Electronic communication in phosphine substituted bridged dirhenium complexes - clarifying ambiguities raised by the redox non-innocence of the C4H2- and C4-bridges.

PubMed

Li, Yan; Blacque, Olivier; Fox, Thomas; Luber, Sandra; Polit, Walther; Winter, Rainer F; Venkatesan, Koushik; Berke, Heinz

2016-04-07

The mononuclear rhenium carbyne complex trans-[Re(C[triple bond, length as m-dash]CSiMe3)([triple bond, length as m-dash]C-Me)(PMe3)4][PF6] (2) was prepared in 90% yield by heating a mixture of the dinitrogen complex trans-[ReCl(N2)(PMe3)4] (1), TlPF6, and an excess of HC[triple bond, length as m-dash]CSiMe3. 2 could be deprotonated with KOtBu to the vinylidene complex trans-[Re(C[triple bond, length as m-dash]CSiMe3)([double bond, length as m-dash]C[double bond, length as m-dash]CH2)(PMe3)4] (3) in 98% yield. Oxidation of 3 with 1.2 equiv. of [Cp2Fe][PF6] at -78 °C gave the Cβ-C'β coupled dinuclear rhenium biscarbyne complex trans-[(Me3SiC[triple bond, length as m-dash]C)(PMe3)4Re[triple bond, length as m-dash]C-CH2-CH2-C[triple bond, length as m-dash]Re(PMe3)4(C[triple bond, length as m-dash]CSiMe3)][PF6]2 (5) in 92% yield. Deprotonation of 5 with an excess of KOtBu in THF produced the diamagnetic trans-[(Me3SiC[triple bond, length as m-dash]C)(PMe3)4Re[double bond, length as m-dash]C[double bond, length as m-dash]CH-CH[double bond, length as m-dash]C[double bond, length as m-dash]Re(PMe3)4(C[triple bond, length as m-dash]CSiMe3)] complex (E-6(S)) in 87% yield with an E-butadienediylidene bridge. Density functional theory (DFT) calculations of E-6(S) confirmed its singlet ground state. The Z-form of 6 (Z-6(S)) could not be observed, which is in accord with its DFT calculated 17.8 kJ mol(-1) higher energy. Oxidation of E-6 with 2 equiv. of [Cp2Fe][PF6] resulted in the stable diamagnetic dicationic trans-[(Me3SiC[triple bond, length as m-dash]C)(PMe3)4Re[triple bond, length as m-dash]C-CH[double bond, length as m-dash]CH-C[triple bond, length as m-dash]Re(PMe3)4(C[triple bond, length as m-dash]CSiMe3)][PF6]2 complex (E-6[PF6]2) with an ethylenylidene dicarbyne structure of the bridge. The paramagnetic mixed-valence (MV) complex E-6[PF6] was obtained by comproportionation of E-6(S) and E-6[PF6]2 or by oxidation of E-6(S) with 1 equiv. of [Cp2Fe][PF6]. The dicationic trans-[(Me3SiC[triple bond, length as m-dash]C)(PMe3)4Re[triple bond, length as m-dash]C-C[triple bond, length as m-dash]C-C[triple bond, length as m-dash]Re(PMe3)4(C[triple bond, length as m-dash]CSiMe3)][PF6]2 (7[PF6]2) complex, attributed a butynedi(triyl) bridge structure, was obtained by deprotonation of E-6[PF6]2 with KOtBu followed by oxidation with 2 equiv. of [Cp2Fe][PF6]. The neutral complex 7 could be accessed best by reduction of 7[PF6]2 with KH in the presence of 18-crown-6. According to DFT calculations 7 possesses two equilibrating electronic states: diamagnetic 7(S) and triplet 7(F) with ferromagnetically coupled spins. The latter is calculated to be 5.2 kcal mol(-1) lower in energy than 7(S). There is experimental evidence that 7(S) prevails in solution. 7 could not be isolated in the crystalline state and is unstable transforming mainly by H-abstraction to give E-6(S). UV-Vis-NIR spectroscopy for the dinuclear rhenium complexes E-6(S), E-6[PF6] and E-6[PF6]2, as well as EPR spectroscopic and variable-temperature magnetization measurements for the MV complex E-6[PF6] were also conducted. Spectro-electrochemical reduction studies on 7[PF6]2 allowed the characterization of the mono- and direduced forms of 7(+) and 7 by means of IR- and UV-Vis-NIR-spectroscopy and revealed the chemical fate of the higher reduced form.

Network Analyses Reveal Pervasive Functional Regulation Between Proteases in the Human Protease Web

PubMed Central

Fortelny, Nikolaus; Cox, Jennifer H.; Kappelhoff, Reinhild; Starr, Amanda E.; Lange, Philipp F.; Pavlidis, Paul; Overall, Christopher M.

2014-01-01

Proteolytic processing is an irreversible posttranslational modification affecting a large portion of the proteome. Protease-cleaved mediators frequently exhibit altered activity, and biological pathways are often regulated by proteolytic processing. Many of these mechanisms have not been appreciated as being protease-dependent, and the potential in unraveling a complex new dimension of biological control is increasingly recognized. Proteases are currently believed to act individually or in isolated cascades. However, conclusive but scattered biochemical evidence indicates broader regulation of proteases by protease and inhibitor interactions. Therefore, to systematically study such interactions, we assembled curated protease cleavage and inhibition data into a global, computational representation, termed the protease web. This revealed that proteases pervasively influence the activity of other proteases directly or by cleaving intermediate proteases or protease inhibitors. The protease web spans four classes of proteases and inhibitors and so links both recently and classically described protease groups and cascades, which can no longer be viewed as operating in isolation in vivo. We demonstrated that this observation, termed reachability, is robust to alterations in the data and will only increase in the future as additional data are added. We further show how subnetworks of the web are operational in 23 different tissues reflecting different phenotypes. We applied our network to develop novel insights into biologically relevant protease interactions using cell-specific proteases of the polymorphonuclear leukocyte as a system. Predictions from the protease web on the activity of matrix metalloproteinase 8 (MMP8) and neutrophil elastase being linked by an inactivating cleavage of serpinA1 by MMP8 were validated and explain perplexing Mmp8 −/− versus wild-type polymorphonuclear chemokine cleavages in vivo. Our findings supply systematically derived and validated evidence for the existence of the protease web, a network that affects the activity of most proteases and thereby influences the functional state of the proteome and cell activity. PMID:24865846

The variable detergent sensitivity of proteases that are utilized for recombinant protein affinity tag removal

PubMed Central

Vergis, James M.; Wiener, Michael C.

2011-01-01

Recombinant proteins typically include one or more affinity tags to facilitate purification and/or detection. Expression constructs with affinity tags often include an engineered protease site for tag removal. Like other enzymes, the activities of proteases can be affected by buffer conditions. The buffers used for integral membrane proteins contain detergents, which are required to maintain protein solubility. We examined the detergent sensitivity of six commonly-used proteases (Enterokinase, Factor Xa, Human Rhinovirus 3C Protease, SUMOstar, Tobacco Etch Virus Protease, and Thrombin) by use of a panel of ninety-four individual detergents. Thrombin activity was insensitive to the entire panel of detergents, thus suggesting it as the optimal choice for use with membrane proteins. Enterokinase and Factor Xa were only affected by a small number of detergents, making them good choices as well. PMID:21539919

The Dash

ERIC Educational Resources Information Center

Buck, Marilyn

2008-01-01

Important events in our lives have a beginning and ending date with a dash in between. What is important is not the dates on either end but what the dash represents. The "dashes" of Amy Morris Homans and Barbara E. Forker are examined. Each touched the lives of their students and the profession that was their lives. Homans understood the…

The Nutritional Characteristics of the Hypotensive WASHOKU-modified DASH Diet: A Sub-analysis of the DASH-JUMP Study.

PubMed

Kawamura, Atsuko; Kajiya, Katsuko; Kishi, Hiroko; Inagaki, Junko; Mitarai, Makoto; Oda, Hiroshi; Umemoto, Seiji; Kobayashi, Sei

2018-01-01

We developed a WASHOKU-modified DASH diet named DASH-JUMP. We previously reported the hypotensive effect of the DASH-JUMP diet in Japanese participants with untreated high-normal Blood Pressure (BP) or stage 1 hypertension. We aim to introduce the DASH-JUMP diet worldwide as a new lifestyle medicine. Accordingly, we prospectively assessed the nutritional characteristics of the DASH-JUMP diet. Participants were treated with the DASH-JUMP diet for 2 months. Then, for 4 months after the intervention, they consumed their usual diets. We conducted a nutritional survey using the FFQg nutrient questionnaire at baseline and after 1, 2, 3, and 6 months. We received completed questionnaires from 55 participants (28 men and 27 women; mean age 54.2 ± 8.0 years) and analyzed them. The DASH-JUMP diet is rich in green-yellow vegetables, seaweed, milk, and mushrooms, while it has low contents of meat, eggs, confectionery, oils and fats, pickles, shellfish boiled in sweetened soy sauce, and fruits. Nutrients significantly associated with the observed change in systolic BP were niacin (P = 0.005) and carbohydrate (P = 0.033). The results of the FFQg questionnaire revealed that participants who had an increased BP at 1 month after ceasing the intervention had eating habits that broadly imitated the DASH-JUMP diet at 4 months after ceasing the intervention. Therefore, the systolic and diastolic BP values at 4 months after ceasing the intervention decreased significantly compared to those at baseline. The DASH-JUMP diet may represent a new lifestyle medicine for reducing hypertension. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

The NCAR Digital Asset Services Hub (DASH): Implementing Unified Data Discovery and Access

NASA Astrophysics Data System (ADS)

Stott, D.; Worley, S. J.; Hou, C. Y.; Nienhouse, E.

2017-12-01

The National Center for Atmospheric Research (NCAR) Directorate created the Data Stewardship Engineering Team (DSET) to plan and implement an integrated single entry point for uniform digital asset discovery and access across the organization in order to improve the efficiency of access, reduce the costs, and establish the foundation for interoperability with other federated systems. This effort supports new policies included in federal funding mandates, NSF data management requirements, and journal citation recommendations. An inventory during the early planning stage identified diverse asset types across the organization that included publications, datasets, metadata, models, images, and software tools and code. The NCAR Digital Asset Services Hub (DASH) is being developed and phased in this year to improve the quality of users' experiences in finding and using these assets. DASH serves to provide engagement, training, search, and support through the following four nodes (see figure). DASH MetadataDASH provides resources for creating and cataloging metadata to the NCAR Dialect, a subset of ISO 19115. NMDEdit, an editor based on a European open source application, has been configured for manual entry of NCAR metadata. CKAN, an open source data portal platform, harvests these XML records (along with records output directly from databases) from a Web Accessible Folder (WAF) on GitHub for validation. DASH SearchThe NCAR Dialect metadata drives cross-organization search and discovery through CKAN, which provides the display interface of search results. DASH search will establish interoperability by facilitating metadata sharing with other federated systems. DASH ConsultingThe DASH Data Curation & Stewardship Coordinator assists with Data Management (DM) Plan preparation and advises on Digital Object Identifiers. The coordinator arranges training sessions on the DASH metadata tools and DM planning, and provides one-on-one assistance as requested. DASH RepositoryA repository is under development for NCAR datasets currently not in existing lab-managed archives. The DASH repository will be under NCAR governance and meet Trustworthy Repositories Audit & Certification (TRAC) requirements. This poster will highlight the processes, lessons learned, and current status of the DASH effort at NCAR.

Synergistic oxygen atom transfer by ruthenium complexes with non-redox metal ions.

PubMed

Lv, Zhanao; Zheng, Wenrui; Chen, Zhuqi; Tang, Zhiming; Mo, Wanling; Yin, Guochuan

2016-07-28

Non-redox metal ions can affect the reactivity of active redox metal ions in versatile biological and heterogeneous oxidation processes; however, the intrinsic roles of these non-redox ions still remain elusive. This work demonstrates the first example of the use of non-redox metal ions as Lewis acids to sharply improve the catalytic oxygen atom transfer efficiency of a ruthenium complex bearing the classic 2,2'-bipyridine ligand. In the absence of Lewis acid, the oxidation of ruthenium(ii) complex by PhI(OAc)2 generates the Ru(iv)[double bond, length as m-dash]O species, which is very sluggish for olefin epoxidation. When Ru(bpy)2Cl2 was tested as a catalyst alone, only 21.2% of cyclooctene was converted, and the yield of 1,2-epoxycyclooctane was only 6.7%. As evidenced by electronic absorption spectra and EPR studies, both the oxidation of Ru(ii) by PhI(OAc)2 and the reduction of Ru(iv)[double bond, length as m-dash]O by olefin are kinetically slow. However, adding non-redox metal ions such as Al(iii) can sharply improve the oxygen transfer efficiency of the catalyst to 100% conversion with 89.9% yield of epoxide under identical conditions. Through various spectroscopic characterizations, an adduct of Ru(iv)[double bond, length as m-dash]O with Al(iii), Ru(iv)[double bond, length as m-dash]O/Al(iii), was proposed to serve as the active species for epoxidation, which in turn generated a Ru(iii)-O-Ru(iii) dimer as the reduced form. In particular, both the oxygen transfer from Ru(iv)[double bond, length as m-dash]O/Al(iii) to olefin and the oxidation of Ru(iii)-O-Ru(iii) back to the active Ru(iv)[double bond, length as m-dash]O/Al(iii) species in the catalytic cycle can be remarkably accelerated by adding a non-redox metal, such as Al(iii). These results have important implications for the role played by non-redox metal ions in catalytic oxidation at redox metal centers as well as for the understanding of the redox mechanism of ruthenium catalysts in the oxygen atom transfer reaction.

Correlation of the degree of clavicle shortening after non-surgical treatment of midshaft fractures with upper limb function.

PubMed

Figueiredo, Gustavo Santiago de Lima; Tamaoki, Marcel Jun Sugawara; Dragone, Bruno; Utino, Artur Yudi; Netto, Nicola Archetti; Matsumoto, Marcelo Hide; Matsunaga, Fábio Teruo

2015-06-17

Despite the use of non-surgical methods to treat for the majority of midshaft fractures of the clavicle, it is remains controversial whether shortening of this bone following non-surgical treatment of a middle third fracture affects upper limb function. We conducted a cohort study by sequentially recruiting 59 patients with a fracture of the middle third of the clavicle. All patients were treated nonsurgically with a figure-of-eight bandage until clinical and radiological findings indicated healing of the fracture. Functional outcome was assessed using the Disability of Arm, Hand and Shoulder (DASH) score revalidated for the Portuguese language, other outcomes assessed included: pain measured by visual analogue scale (VAS); radiographies to measure the degree of shortening, fracture consolidation and fracture malunion. Information were also collected regarding the mechanism of injury, patient's daily activities level and epidemiological features of the patient cohort. The results of our findings are expressed as the comparison of the functional outcome with the degree of shortening. Patients were assessed six weeks and one year after injury. In the first evaluation, the mean DASH score was 28.84 and pain measured by VAS was 2.57. In the second evaluation (one year after injury) the mean DASH score was 8.18 and pain was 0.84. The mean clavicle shortening was 0.92 cm, ranging from 0 to 3 cm (SD = 0.64). There were no correlation between the degree of shortening and DASH score after six weeks and one year (p = 0.073 and 0.706, respectively). When only patients with of shortening greater than 2 cm were assessed for correlation, the result did not change. We conclude that clavicle shortening after nonsurgical treatment with a figure-of-eight bandage does not affect limb function, even when shortening exceeds 2 cm. ISRCTN85206617 . Registered 12 May 2014.

Inhibition of melanosome transfer results in skin lightening.

PubMed

Seiberg, M; Paine, C; Sharlow, E; Andrade-Gordon, P; Costanzo, M; Eisinger, M; Shapiro, S S

2000-08-01

The chemical basis of melanogenesis is well documented, but the mechanism of melanosome transfer and the regulation of pigmentation by keratinocyte-melanocyte interactions are not well understood. Therefore we examined the effects of serine protease inhibitors on skin pigmentation and found that the protease-activated receptor 2, expressed on keratinocytes, may regulate pigmentation via keratinocyte-melanocyte interactions. Here we show that modulation of protease-activated receptor 2 activation affects melanosome transfer into keratinocytes, resulting in changes in pigment production and deposition. SLIGRL, the protease-activated receptor 2 activating peptide, enhanced melanosome ingestion by keratinocytes, thus increasing pigment deposition. RWJ-50353, a serine protease inhibitor, led to reduced pigment deposition in melanocytes and depigmentation. Electron microscopy studies illustrated an accumulation of immature melanosomes inside melanocytes and abnormal dendrite dynamics in RWJ-50353-treated epidermal equivalents. RWJ-50353 induced a visible and dose-dependent skin lightening effect in the dark-skinned Yucatan swine. Examinations by electron microscopy indicated that the in vivo transfer of melanosomes from melanocytes to keratinocytes was affected. Our data suggest that modulation of keratinocyte-melanocyte interactions via the protease-activated receptor 2 pathway affects melanosome transfer. The use of RWJ-50353 to modulate protease-activated receptor 2 activation could lead to a new class of depigmenting agents.

Measurement properties of the QuickDASH (disabilities of the arm, shoulder and hand) outcome measure and cross-cultural adaptations of the QuickDASH: a systematic review.

PubMed

Kennedy, Carol A; Beaton, Dorcas E; Smith, Peter; Van Eerd, Dwayne; Tang, Kenneth; Inrig, Taucha; Hogg-Johnson, Sheilah; Linton, Denise; Couban, Rachel

2013-11-01

To identify and synthesize evidence for the measurement properties of the QuickDASH, a shortened version of the 30-item DASH (Disabilities of the Arm, Shoulder and Hand) instrument. This systematic review used a best evidence synthesis approach to critically appraise the measurement properties [using COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN)] of the QuickDASH and cross-cultural adaptations. A standard search strategy was conducted between 2005 (year of first publication of QuickDASH) and March 2011 in MEDLINE, EMBASE and CINAHL. The search identified 14 studies to include in the best evidence synthesis of the QuickDASH. A further 11 studies were identified on eight cross-cultural adaptation versions. Many measurement properties of the QuickDASH have been evaluated in multiple studies and across most of the measurement properties. The best evidence synthesis of the QuickDASH English version suggests that this tool is performing well with strong positive evidence for reliability and validity (hypothesis testing), and moderate positive evidence for structural validity testing. Strong negative evidence was found for responsiveness due to lower correlations with global estimates of change. Information about the measurement properties of the cross-cultural adaptation versions is still lacking, or the available information is of poor overall methodological quality.

Some transition metal complexes derived from mono- and di-ethynyl perfluorobenzenes.

PubMed

Armitt, David J; Bruce, Michael I; Gaudio, Maryka; Zaitseva, Natasha N; Skelton, Brian W; White, Allan H; Le Guennic, Boris; Halet, Jean-François; Fox, Mark A; Roberts, Rachel L; Hartl, Frantisek; Low, Paul J

2008-12-21

Transition metal alkynyl complexes containing perfluoroaryl groups have been prepared directly from trimethylsilyl-protected mono- and di-ethynyl perfluoroarenes by simple desilylation/metallation reaction sequences. Reactions between Me(3)SiC[triple bond, length as m-dash]CC(6)F(5) and RuCl(dppe)Cp' [Cp' = Cp, Cp*] in the presence of KF in MeOH give the monoruthenium complexes Ru(C[triple bond, length as m-dash]CC(6)F(5))(dppe)Cp' [Cp' = Cp (); Cp* ()], which are related to the known compound Ru(C[triple bond, length as m-dash]CC(6)F(5))(PPh(3))(2)Cp (). Treatment of Me(3)SiC[triple bond, length as m-dash]CC(6)F(5) with Pt(2)(mu-dppm)(2)Cl(2) in the presence of NaOMe in MeOH gave the bis(alkynyl) complex Pt(2)(mu-dppm)(2)(C[triple bond, length as m-dash]CC(6)F(5))(2) (). The Pd(0)/Cu(i)-catalysed reactions between Au(C[triple bond, length as m-dash]CC(6)F(5))(PPh(3)) and Mo( identical withCBr)(CO)(2)Tp* [Tp* = hydridotris(3.5-dimethylpyrazoyl)borate], Co(3)(mu(3)-CBr)(mu-dppm)(CO)(7) or IC[triple bond, length as m-dash]CFc [Fc = (eta(5)-C(5)H(4))FeCp] afford Mo( identical withCC[triple bond, length as m-dash]CC(6)F(5))(CO)(2)Tp* (), Co(3)(mu(3)-CC[triple bond, length as m-dash]CC(6)F(5))(mu-dppm)(CO)(7) () and FcC[triple bond, length as m-dash]CC[triple bond, length as m-dash]CC(6)F(5) (), respectively. The diruthenium complexes 1,4-{Cp'(PP)RuC[triple bond, length as m-dash]C}(2)C(6)F(4) [(PP)Cp' = (PPh(3))(2)Cp (); (dppe)Cp (); (dppe)Cp* ()] are prepared from 1,4-(Me(3)SiC[triple bond, length as m-dash]C)(2)C(6)F(4) in a manner similar to that described for the monoruthenium complexes -. The non-fluorinated complexes 1,4-{Cp'(PP)RuC[triple bond, length as m-dash]C}(2)C(6)H(4) [(PP)Cp' = (PPh(3))(2)Cp (); (dppe)Cp (); (dppe)Cp* ()], prepared for comparison, are obtained from 1,4-(Me(3)SiC[triple bond, length as m-dash]C)(2)C(6)H(4). Spectro-electrochemical studies of the ruthenium aryl and arylene alkynyl complexes - and -, together with DFT-based computational studies on suitable model systems, indicate that perfluorination of the aromatic ring has little effect on the electronic structures of these compounds, and that the frontier orbitals have appreciable diethynylphenylene character. Molecular structure determinations are reported for the fluoroaromatic complexes , , , and .

The PROMIS physical function correlates with the QuickDASH in patients with upper extremity illness.

PubMed

Overbeek, Celeste L; Nota, Sjoerd P F T; Jayakumar, Prakash; Hageman, Michiel G; Ring, David

2015-01-01

To assess disability more efficiently with less burden on the patient, the National Institutes of Health has developed the Patient Reported Outcomes Measurement Information System (PROMIS) Physical Function-an instrument based on item response theory and using computer adaptive testing (CAT). Initially, upper and lower extremity disabilities were not separated and we were curious if the PROMIS Physical Function CAT could measure upper extremity disability and the Quick Disability of Arm, Shoulder and Hand (QuickDASH). We aimed to find correlation between the PROMIS Physical Function and the QuickDASH questionnaires in patients with upper extremity illness. Secondarily, we addressed whether the PROMIS Physical Function and QuickDASH correlate with the PROMIS Depression CAT and PROMIS Pain Interference CAT instruments. Finally, we assessed factors associated with QuickDASH and PROMIS Physical Function in multivariable analysis. A cohort of 93 outpatients with upper extremity illnesses completed the QuickDASH and three PROMIS CAT questionnaires: Physical Function, Pain Interference, and Depression. Pain intensity was measured with an 11-point ordinal measure (0-10 numeric rating scale). Correlation between PROMIS Physical Function and the QuickDASH was assessed. Factors that correlated with the PROMIS Physical Function and QuickDASH were assessed in multivariable regression analysis after initial bivariate analysis. There was a moderate correlation between the PROMIS Physical Function and the QuickDASH questionnaire (r=-0.55, p<0.001). Greater disability as measured with the PROMIS and QuickDASH correlated most strongly with PROMIS Depression (r=-0.35, p<0.001 and r=0.34, p<0.001 respectively) and Pain Interference (r=-0.51, p<0.001 and r=0.74, p<0.001 respectively). The factors accounting for the variability in PROMIS scores are comparable to those for the QuickDASH except that the PROMIS Physical Function is influenced by other pain conditions while the QuickDASH is not. The PROMIS Physical Function instrument may be used as an upper extremity disability measure, as it correlates with the QuickDASH questionnaire, and both instruments are influenced most strongly by the degree to which pain interferes with achieving goals. Level III, diagnostic study. See the Instructions for Authors for a complete description of levels of evidence.

The Effects of Cold Whirlpool on Power, Speed, Agility, and Range of Motion

PubMed Central

Patterson, Stephen M.; Udermann, Brian E.; Doberstein, Scott T.; Reineke, David M.

2008-01-01

The purpose was to determine if cold whirlpool treatment decreases functional performance equally regardless of gender. A secondary aim was to determine if there is a gradual increase in functional performance across time. Twenty-one college-aged subjects volunteered to participate in this study and were required to perform four measures of functional performance including: counter movement vertical jump, T-test, 36.58-meter dash (40-yard), and active range of motion of the ankle. Participants were treated with a 20 minute, 10 degree Celsius cold whirlpool following the pre-test of a given functional performance measure. Participants demonstrated significant decreases in counter movement vertical jump, T-test, and 40-yard dash performance immediately following treatment. Vertical jump performance remained impaired for at least 32 minutes. While both the T-test and 40-yard dash were affected for 7 and 22 minutes post- treatment, respectively. Participants also demonstrated significant decreases in peak power and average power immediately after and for 32 minutes post-treatment. Dorsiflexion was significantly decreased 7 and 12 minutes following treatment. There were no differences for plantar flexion, inversion, or eversion. These data suggest functional performance was affected immediately following and for up to 32 minutes after cold whirlpool treatment. It was also evident that there is a gradual performance increase for each measure of functional performance across time. Therefore, the consequences should be carefully considered before returning athletes to activity following cold whirlpool treatment. Key pointsCryotherapy is a common and highly effective modality in treating acute and chronic athletic injuries.The results indicated that cold whirlpool does have an immediate and subsequent effect on functional performance.Understanding how cold whirlpool adversely affects functional performance allows clinicians to continue using this modality before vigorous athletic activity. PMID:24149907

Greater accordance with the Dietary Approaches to Stop Hypertension dietary pattern is associated with lower diet-related greenhouse gas production but higher dietary costs in the United Kingdom12

PubMed Central

Monsivais, Pablo; Scarborough, Peter; Lloyd, Tina; Mizdrak, Anja; Luben, Robert; Mulligan, Angela A; Wareham, Nicholas J; Woodcock, James

2015-01-01

Background: The Dietary Approaches to Stop Hypertension (DASH) diet is a proven way to prevent and control hypertension and other chronic disease. Because the DASH diet emphasizes plant-based foods, including vegetables and grains, adhering to this diet might also bring about environmental benefits, including lower associated production of greenhouse gases (GHGs). Objective: The objective was to examine the interrelation between dietary accordance with the DASH diet and associated GHGs. A secondary aim was to examine the retail cost of diets by level of DASH accordance. Design: In this cross-sectional study of adults aged 39–79 y from the European Prospective Investigation into Cancer and Nutrition–Norfolk, United Kingdom cohort (n = 24,293), dietary intakes estimated from food-frequency questionnaires were analyzed for their accordance with the 8 DASH food and nutrient-based targets. Associations between DASH accordance, GHGs, and dietary costs were evaluated in regression analyses. Dietary GHGs were estimated with United Kingdom-specific data on carbon dioxide equivalents associated with commodities and foods. Dietary costs were estimated by using national food prices from a United Kingdom–based supermarket comparison website. Results: Greater accordance with the DASH dietary targets was associated with lower GHGs. Diets in the highest quintile of accordance had a GHG impact of 5.60 compared with 6.71 kg carbon dioxide equivalents/d for least-accordant diets (P < 0.0001). Among the DASH food groups, GHGs were most strongly and positively associated with meat consumption and negatively with whole-grain consumption. In addition, higher accordance with the DASH diet was associated with higher dietary costs, with the mean cost of diets in the top quintile of DASH scores 18% higher than that of diets in the lowest quintile (P < 0.0001). Conclusions: Promoting wider uptake of the DASH diet in the United Kingdom may improve population health and reduce diet-related GHGs. However, to make the DASH diet more accessible, food affordability, particularly for lower income groups, will have to be addressed. PMID:25926505

Greater accordance with the Dietary Approaches to Stop Hypertension dietary pattern is associated with lower diet-related greenhouse gas production but higher dietary costs in the United Kingdom.

PubMed

Monsivais, Pablo; Scarborough, Peter; Lloyd, Tina; Mizdrak, Anja; Luben, Robert; Mulligan, Angela A; Wareham, Nicholas J; Woodcock, James

2015-07-01

The Dietary Approaches to Stop Hypertension (DASH) diet is a proven way to prevent and control hypertension and other chronic disease. Because the DASH diet emphasizes plant-based foods, including vegetables and grains, adhering to this diet might also bring about environmental benefits, including lower associated production of greenhouse gases (GHGs). The objective was to examine the interrelation between dietary accordance with the DASH diet and associated GHGs. A secondary aim was to examine the retail cost of diets by level of DASH accordance. In this cross-sectional study of adults aged 39-79 y from the European Prospective Investigation into Cancer and Nutrition-Norfolk, United Kingdom cohort (n = 24,293), dietary intakes estimated from food-frequency questionnaires were analyzed for their accordance with the 8 DASH food and nutrient-based targets. Associations between DASH accordance, GHGs, and dietary costs were evaluated in regression analyses. Dietary GHGs were estimated with United Kingdom-specific data on carbon dioxide equivalents associated with commodities and foods. Dietary costs were estimated by using national food prices from a United Kingdom-based supermarket comparison website. Greater accordance with the DASH dietary targets was associated with lower GHGs. Diets in the highest quintile of accordance had a GHG impact of 5.60 compared with 6.71 kg carbon dioxide equivalents/d for least-accordant diets (P < 0.0001). Among the DASH food groups, GHGs were most strongly and positively associated with meat consumption and negatively with whole-grain consumption. In addition, higher accordance with the DASH diet was associated with higher dietary costs, with the mean cost of diets in the top quintile of DASH scores 18% higher than that of diets in the lowest quintile (P < 0.0001). Promoting wider uptake of the DASH diet in the United Kingdom may improve population health and reduce diet-related GHGs. However, to make the DASH diet more accessible, food affordability, particularly for lower income groups, will have to be addressed.

Adherence to the Dietary Approaches to Stop Hypertension (DASH) diet in relation to obesity among Iranian female nurses.

PubMed

Barak, Farzaneh; Falahi, Ebrahim; Keshteli, Ammar Hassanzadeh; Yazdannik, Ahmadreza; Esmaillzadeh, Ahmad

2015-03-01

Limited observational studies have considered habitual consumption of the general population to examine the relationship between the Dietary Approaches to Stop Hypertension (DASH) diet and obesity. The aim of the present study was to investigate adherence to the DASH diet in relation to general and central obesity among female nurses in Isfahan, Iran. Cross-sectional study carried out among 293 female nurses aged >30 years who were selected by a multistage, cluster random sampling method. Usual dietary intakes were assessed using a validated FFQ. We constructed the DASH score based on foods and nutrients emphasized or minimized in the DASH diet, focusing on eight components: high intake of fruits, vegetables, nuts and legumes, low-fat dairy products and whole grains and low intakes of sodium, sweetened beverages, and red and processed meats. General and abdominal obesity were defined as BMI ≥ 25 kg/m2 and waist circumference ≥ 88 cm, respectively. Isfahan, Iran. Female nurses (n 293) aged >30 years. Increased adherence to the DASH diet was associated with older age (P<0.01) and lower waist circumference (P=0.04). There was no statistically significant difference in the prevalence of general obesity between extreme quartiles of the DASH diet score. After adjustment for age, energy intake and other confounding factors, DASH diet score was not significantly associated with obesity. However, with further controlling for other dietary factors, those in the highest quartile of DASH diet score were 71 % less likely to have general obesity compared with those in the lowest quartile. In addition, following a DASH diet was inversely associated with central obesity after adjustment for potential confounders (OR=0.37; 95 % CI 0.14, 0.96). We found that adherence to the DASH diet was inversely related to central obesity among Iranian adult females. This association remained significant even after adjustment for potential confounders.

Outcomes of nonoperatively treated displaced scapular body fractures.

PubMed

Dimitroulias, Apostolos; Molinero, Kenneth G; Krenk, Daniel E; Muffly, Matthew T; Altman, Daniel T; Altman, Gregory T

2011-05-01

Displaced scapular body fractures most commonly are treated conservatively. However there is conflicting evidence in the literature regarding the outcomes owing to retrospective design of studies, different classification systems, and diverse outcome tools. The functional outcome after nonoperative management of displaced scapular body fractures was assessed by change in the DASH (Disability of Arm, Shoulder and Hand) score; (2) the radiographic outcome was assessed by the change of the glenopolar angle (GPA); and (3) associated scapular and extrascapular injuries that may affect outcome were identified. Forty-nine consecutive patients were treated with early passive and active ROM exercises for a displaced scapular body fracture. We followed 32 of these patients (65.3%) for a minimum of 6 months (mean, 15 months; range, 6-33 months). Mean age of the patients was 46.9 years (range, 21-84 years) and the mean Injury Severity Score (ISS) was 21.5 (range, 5-50). Subjective functional results (DASH score) and radiographic assessment (fracture union, glenopolar angle) were measured. All fractures healed uneventfully. The mean change of glenopolar angle was 9° (range, 0°-20°). The mean change of the DASH score was 10.2, which is a change with minimal clinical importance. There was a correlation between the change in this score with the ISS and presence of rib fractures. Satisfactory outcomes are reported with nonoperative treatment of displaced scapular body fractures. We have shown that the severity of ISS and the presence of rib fractures adversely affect the clinical outcome.

The Mad Dash To Compute.

ERIC Educational Resources Information Center

Healy, Jane M.

1999-01-01

Discusses trade-offs and ramifications of technology use in schools. Cutbacks in proven staples of mental development (arts, music, drama, and physical education) are used to finance technology programs. Youngsters often use educational software for mindless fun. Few advocates consider how extended computer usage affects children's developing…

OHS Provides Easy Heart Health Tips at “DASH and Dine” Event | Poster

Cancer.gov

The lunchtime dash to a nearby eatery is a daily ritual at NCI at Frederick, but Occupational Health Services (OHS) recently showed employees a different, healthy kind of DASH—Dietary Approaches to Stop Hypertension. Employees who visited the educational lunch-hour event, called “DASH and Dine,” had the chance to speak with OHS staff about the benefits of the DASH Eating Plan,

DASH Diet: Reducing Hypertension through Diet and Lifestyle

MedlinePlus

... and Cardiovascular Health DASH Diet DASH Diet: Reducing Hypertension through Diet and Lifestyle Reviewed by Taylor Wolfram, ... vkusidey/iStock/Thinkstock Think you can't stop hypertension, or high blood pressure? You might be able ...

Extracellular Protease Inhibition Alters the Phenotype of Chondrogenically Differentiating Human Mesenchymal Stem Cells (MSCs) in 3D Collagen Microspheres.

PubMed

Han, Sejin; Li, Yuk Yin; Chan, Barbara Pui

2016-01-01

Matrix remodeling of cells is highly regulated by proteases and their inhibitors. Nevertheless, how would the chondrogenesis of mesenchymal stem cells (MSCs) be affected, when the balance of the matrix remodeling is disturbed by inhibiting matrix proteases, is incompletely known. Using a previously developed collagen microencapsulation platform, we investigated whether exposing chondrogenically differentiating MSCs to intracellular and extracellular protease inhibitors will affect the extracellular matrix remodeling and hence the outcomes of chondrogenesis. Results showed that inhibition of matrix proteases particularly the extracellular ones favors the phenotype of fibrocartilage rather than hyaline cartilage in chondrogenically differentiating hMSCs by upregulating type I collagen protein deposition and type II collagen gene expression without significantly altering the hypertrophic markers at gene level. This study suggests the potential of manipulating extracellular proteases to alter the outcomes of hMSC chondrogenesis, contributing to future development of differentiation protocols for fibrocartilage tissues for intervertebral disc and meniscus tissue engineering.

Extracellular Protease Inhibition Alters the Phenotype of Chondrogenically Differentiating Human Mesenchymal Stem Cells (MSCs) in 3D Collagen Microspheres

PubMed Central

Han, Sejin; Li, Yuk Yin; Chan, Barbara Pui

2016-01-01

Matrix remodeling of cells is highly regulated by proteases and their inhibitors. Nevertheless, how would the chondrogenesis of mesenchymal stem cells (MSCs) be affected, when the balance of the matrix remodeling is disturbed by inhibiting matrix proteases, is incompletely known. Using a previously developed collagen microencapsulation platform, we investigated whether exposing chondrogenically differentiating MSCs to intracellular and extracellular protease inhibitors will affect the extracellular matrix remodeling and hence the outcomes of chondrogenesis. Results showed that inhibition of matrix proteases particularly the extracellular ones favors the phenotype of fibrocartilage rather than hyaline cartilage in chondrogenically differentiating hMSCs by upregulating type I collagen protein deposition and type II collagen gene expression without significantly altering the hypertrophic markers at gene level. This study suggests the potential of manipulating extracellular proteases to alter the outcomes of hMSC chondrogenesis, contributing to future development of differentiation protocols for fibrocartilage tissues for intervertebral disc and meniscus tissue engineering. PMID:26760956

A comparison of upper limb amputees and patients with upper limb injuries using the Disability of the Arm, Shoulder and Hand (DASH).

PubMed

Davidson, Judith

To use the Disability of the Arm Shoulder and Hand (DASH) scale to measure the disability of patients with upper limb amputation(s) and to compare these to other upper limb injuries. All 274 patients over the age of 18 years presenting to Prince Henry Hospital in Sydney over a 4-year time frame were given the DASH assessment tool and asked to complete it under supervision of the Occupational Therapist. Patients with brachial plexus injuries, Complex Regional Pain Syndrome and bilateral upper limb amputations demonstrated significantly higher levels of disability to patients with unilateral upper limb amputations. Partial hand amputees reported a higher level of disability than major unilateral upper limb amputees. For the 48 patients who completed pre- and post-treatment assessments, there was a significant improvement in their health status. Further research is required to understand the factors that affect a patient's perceptions of their disability. Perhaps the definitive nature of an amputation and the immediate involvement of highly skilled health professionals serve to assist patients to accept their injury and therefore minimizes the level of disability.

The cross-cultural adaptation of the DASH questionnaire in Thai (DASH-TH).

PubMed

Tongprasert, Siam; Rapipong, Jeeranan; Buntragulpoontawee, Montana

2014-01-01

Clinical measurement. Currently there are no self-report questionnaires in Thai to evaluate disability levels in patients suffering from upper extremity musculoskeletal disorders. To translate and cross-cultural adaptation the disabilities of the arm, shoulder and hand (DASH) questionnaire to Thai version and to evaluate content validity, construct validity and internal consistency of the questionnaire. The DASH-TH was produced by following cross-cultural adaptation guidelines stated by the Institute for Work and Health (IWH). Forty Thai patients with arm, shoulder or hand problems participated in field testing of the questionnaire. Content validity was determined by obtaining the item-objective congruence (IOC) value for each questionnaire item. Correlation between the DASH-TH score and numeric rating scale was used to assess construct validity. Internal consistency of DASH-TH was measured using Cronbach's alpha coefficient. Forty patients (14 males, 26 females) with arm, shoulder or hand problems enrolled in the present study. The average age of patients was 44.8 years. The index of item-objective congruence (IOC) of each item ranged from 0.7 to 1.0. The Cronbach's alpha coefficient of the questionnaire was 0.938. There was no correlation between DASH-TH score and numeric rating scale. The DASH-TH has high content validity and internal consistency. N/A. Copyright © 2014 Hanley & Belfus. Published by Elsevier Inc. All rights reserved.

Dietary approaches to stop hypertension diet and obesity: A cross-sectional study of Iranian children and adolescents.

PubMed

Golpour-Hamedani, Sahar; Mohammadifard, Noushin; Khosravi, Alireza; Feizi, Awat; Safavi, Sayyed Morteza

2017-01-01

Few studies have investigated the effects of dietary approaches to stop hypertension (DASH) diet on obesity in children. The present study was conducted to examine adherence to the DASH diet in relation to obesity in children and adolescents, Isfahan, Iran. A cross-sectional study was carried out among 456 children aged 11-18 years who were selected by random cluster sampling method. Dietary intakes were assessed using a validated Food Frequency Questionnaire (FFQ). The DASH score was constructed based on food items emphasized or minimized in the DASH diet. Anthropometric measurements were conducted based on standard protocols. General and abdominal obesity were defined based on body mass index ≥ 95th percentiles and waist: height ratio of more than 0.5, respectively. Higher adherence to DASH diet was inversely associated with general obesity (odds ratioT1 vs. T3 3.34, 95% confidence interval 1.28-8.75); however, after controlling for confounding factors, this association disappeared. Furthermore, higher adherence to DASH diet was negatively associated with central obesity in children, but the relation was not statistically significant. We concluded that there was an inverse nonsignificant association between adherence to DASH diet and general obesity indices after adjustment for potential confounders. Further, well-designed randomized clinical trial studies are suggested to find out the effect of DASH diet on obesity obviously.

Murine Coronavirus Ubiquitin-Like Domain Is Important for Papain-Like Protease Stability and Viral Pathogenesis

PubMed Central

Mielech, Anna M.; Deng, Xufang; Chen, Yafang; Kindler, Eveline; Wheeler, Dorthea L.; Mesecar, Andrew D.; Thiel, Volker; Perlman, Stanley

2015-01-01

ABSTRACT Ubiquitin-like domains (Ubls) now are recognized as common elements adjacent to viral and cellular proteases; however, their function is unclear. Structural studies of the papain-like protease (PLP) domains of coronaviruses (CoVs) revealed an adjacent Ubl domain in severe acute respiratory syndrome CoV, Middle East respiratory syndrome CoV, and the murine CoV, mouse hepatitis virus (MHV). Here, we tested the effect of altering the Ubl adjacent to PLP2 of MHV on enzyme activity, viral replication, and pathogenesis. Using deletion and substitution approaches, we identified sites within the Ubl domain, residues 785 to 787 of nonstructural protein 3, which negatively affect protease activity, and valine residues 785 and 787, which negatively affect deubiquitinating activity. Using reverse genetics, we engineered Ubl mutant viruses and found that AM2 (V787S) and AM3 (V785S) viruses replicate efficiently at 37°C but generate smaller plaques than wild-type (WT) virus, and AM2 is defective for replication at higher temperatures. To evaluate the effect of the mutation on protease activity, we purified WT and Ubl mutant PLP2 and found that the proteases exhibit similar specific activities at 25°C. However, the thermal stability of the Ubl mutant PLP2 was significantly reduced at 30°C, thereby reducing the total enzymatic activity. To determine if the destabilizing mutation affects viral pathogenesis, we infected C57BL/6 mice with WT or AM2 virus and found that the mutant virus is highly attenuated, yet it replicates sufficiently to elicit protective immunity. These studies revealed that modulating the Ubl domain adjacent to the PLP reduces protease stability and viral pathogenesis, revealing a novel approach to coronavirus attenuation. IMPORTANCE Introducing mutations into a protein or virus can have either direct or indirect effects on function. We asked if changes in the Ubl domain, a conserved domain adjacent to the coronavirus papain-like protease, altered the viral protease activity or affected viral replication or pathogenesis. Our studies using purified wild-type and Ubl mutant proteases revealed that mutations in the viral Ubl domain destabilize and inactivate the adjacent viral protease. Furthermore, we show that a CoV encoding the mutant Ubl domain is unable to replicate at high temperature or cause lethal disease in mice. Our results identify the coronavirus Ubl domain as a novel modulator of viral protease stability and reveal manipulating the Ubl domain as a new approach for attenuating coronavirus replication and pathogenesis. PMID:25694594

Adherence to the DASH diet in relation to psychological profile of Iranian adults.

PubMed

Valipour, Ghazaleh; Esmaillzadeh, Ahmad; Azadbakht, Leila; Afshar, Hamid; Hassanzadeh, Ammar; Adibi, Peyman

2017-02-01

Although empirically derived dietary patterns have been examined in relation to depression, limited data are available linking theory-based dietary patterns and psychological health. We aimed to investigate the association between adherence to DASH-style diet and psychological health among Iranian adults. This cross-sectional study was done among 3846 general public adults in Isfahan, Iran. Dietary assessment was conducted using a validated 106-item dish-based semiquantitative food frequency questionnaire. To investigate participants' adherence to DASH-style diet, we created DASH score based on earlier publications focusing on eight components (fruits, vegetables, nuts and legumes, dairy products, grains, sweetened beverages and sweets, sodium, and red and processed meats). Participants were classified into three categories based on their DASH score [low (≤40), moderate (41-50), and high adherence (≥51)]. This categorization, instead of distribution-based classification, was used due to low adherence to the DASH dietary pattern in the study population. Psychological health was examined by means of validated Hospital Anxiety and Depression Scale and General Health Questionnaire. Depression, anxiety, and psychological distress were defined based on standard criteria. We found that moderate adherence to DASH-style diet was associated with lower odds of depression (OR 0.73; 95 % CI 0.59-0.90, P trend  = 0.63) compared with those with the lowest adherence. In our stratified analyses, these associations remained significant for women (0.70; 0.54-0.91) and for normal-weight participants (0.70; 0.52-0.92). Moreover, after controlling for potential confounders, an inverse association was observed between high adherence to DASH-style diet and anxiety in normal-weight participants (0.61; 0.37-0.98). Such associations were also seen between moderate adherence to DASH-eating style and anxiety in overweight or obese individuals (0.63; 0.42-0.95). We failed to find any significant association between consumption of DASH-style diet and psychological distress. We found an inverse association between moderate adherence to DASH dietary pattern and depression. Further prospective studies are required to confirm these findings.

Indispensable Role of Proteases in Plant Innate Immunity.

PubMed

Balakireva, Anastasia V; Zamyatnin, Andrey A

2018-02-23

Plant defense is achieved mainly through the induction of microbe-associated molecular patterns (MAMP)-triggered immunity (MTI), effector-triggered immunity (ETI), systemic acquired resistance (SAR), induced systemic resistance (ISR), and RNA silencing. Plant immunity is a highly complex phenomenon with its own unique features that have emerged as a result of the arms race between plants and pathogens. However, the regulation of these processes is the same for all living organisms, including plants, and is controlled by proteases. Different families of plant proteases are involved in every type of immunity: some of the proteases that are covered in this review participate in MTI, affecting stomatal closure and callose deposition. A large number of proteases act in the apoplast, contributing to ETI by managing extracellular defense. A vast majority of the endogenous proteases discussed in this review are associated with the programmed cell death (PCD) of the infected cells and exhibit caspase-like activities. The synthesis of signal molecules, such as salicylic acid, jasmonic acid, and ethylene, and their signaling pathways, are regulated by endogenous proteases that affect the induction of pathogenesis-related genes and SAR or ISR establishment. A number of proteases are associated with herbivore defense. In this review, we summarize the data concerning identified plant endogenous proteases, their effect on plant-pathogen interactions, their subcellular localization, and their functional properties, if available, and we attribute a role in the different types and stages of innate immunity for each of the proteases covered.

Purification and characterization of an alkaline protease Prot 1 from Botrytis cinerea : biodetergent catalyst assay.

PubMed

Abidi, Ferid; Limam, Ferid; Marzouki, M Nejib

2007-01-01

Alkaline thiol protease named Prot 1 was isolated from a culture filtrate of Botrytis cinerea. The enzyme was purified by ammonium sulfate fractionation, gel filtration, and ion-exchange chromatography. Thus, the enzyme was purified to homogeneity with specific activity of 30-fold higher than that of the crude broth. The purified alkaline protease has an apparent molecular mass of 43 kDa under denaturing conditions as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The native molecular mass (45 kDa), determined by gel filtration, indicated that the alkaline protease has a monomeric form. The purified protease was biochemically characterized. The enzyme is active at alkaline pH and has a suitable and high thermostability. The optimal pH and temperature for activity were 9.0-10.0 and 60 degrees C, respectively. This protease was stable between pH 5.0 and 12.0. The enzyme retained 85% of its activity by treatment at 50 degrees C over 120 min; it maintained 50% of activity after 60 min of heating at 60 degrees C. Furthermore, the protease retained almost complete activity after 4 wk storage at 25 degrees C. The activity was significantly affected by thiol protease inhibitors, suggesting that the enzyme belongs to the alkaline thiol protease family. With the aim on industrial applications, we focused on studying the stability of the protease in several conditions. Prot 1 activity was not affected by ionic strength and different detergent additives, and, thus, the protease shows remarkable properties as a biodetergent catalyst.

Assessing physical function in adult acquired major upper-limb amputees by combining the Disabilities of the Arm, Shoulder and Hand (DASH) Outcome Questionnaire and clinical examination.

PubMed

Ostlie, Kristin; Franklin, Rosemary J; Skjeldal, Ola H; Skrondal, Anders; Magnus, Per

2011-10-01

To describe physical function in adult acquired major upper-limb amputees (ULAs) by combining self-assessed arm function and physical measures obtained by clinical examinations; to estimate associations between background factors and self-assessed arm function in ULAs; and to assess whether clinical examination findings may be used to detect reduced arm function in unilateral ULAs. postal questionnaires and clinical examinations. Norwegian ULA population. Clinical examinations performed at 3 clinics. Questionnaires: population-based sample (n=224; 57.4% response rate). Clinical examinations: combined referred sample and convenience sample of questionnaire responders (n=70; 83.3% of those invited). SURVEY inclusion criteria: adult acquired major upper-limb amputation, resident in Norway, mastering of spoken and written Norwegian. Not applicable. The Disabilities of the Arm, Shoulder and Hand (DASH) Outcome Questionnaire, and clinical examination of joint motion and muscle strength with and without prostheses. Mean DASH score was 22.7 (95% confidence interval [CI], 20.3-25.0); in bilateral amputees, 35.7 (95% CI, 23.0-48.4); and in unilateral amputees, 22.1 (95% CI, 19.8-24.5). A lower unilateral DASH score (better function) was associated with paid employment (vs not in paid employment: adjusted regression coefficient [aB]=-5.40, P=.033; vs students: aB=-13.88, P=.022), increasing postamputation time (aB=-.27, P=.001), and Norwegian ethnicity (aB=-14.45, P<.001). At clinical examination, we found a high frequency of impaired neck mobility and varying frequencies of impaired joint motion and strength at the shoulder, elbow, and forearm level. Prosthesis wear was associated with impaired joint motion in all upper-limb joints (P<.006) and with reduced shoulder abduction strength (P=.002). Impaired without-prosthesis joint motion in shoulder flexion (ipsilateral: aB=12.19, P=.001) and shoulder abduction (ipsilateral: aB=12.01, P=.005; contralateral: aB=28.82, P=.004) was associated with increased DASH scores. Upper-limb loss clearly affects physical function. DASH score limitation profiles may be useful in individual clinical assessments. Targeted clinical examination may indicate patients with extra rehabilitational needs. Such examinations may be of special importance in relation to prosthesis function. Copyright © 2011 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.

Effects of the dietary approaches to stop hypertension diet alone and in combination with exercise and caloric restriction on insulin sensitivity and lipids.

PubMed

Blumenthal, James A; Babyak, Michael A; Sherwood, Andrew; Craighead, Linda; Lin, Pao-Hwa; Johnson, Julie; Watkins, Lana L; Wang, Jenny T; Kuhn, Cynthia; Feinglos, Mark; Hinderliter, Alan

2010-05-01

This study examined the effects of the Dietary Approaches to Stop Hypertension (DASH) diet on insulin sensitivity and lipids. In a randomized control trial, 144 overweight (body mass index: 25 to 40) men (n=47) and women (n=97) with high blood pressure (130 to 159/85 to 99 mm Hg) were randomly assigned to one of the following groups: (1) DASH diet alone; (2) DASH diet with aerobic exercise and caloric restriction; or (3) usual diet controls (UC). Body composition, fitness, insulin sensitivity, and fasting lipids were measured before and after 4 months of treatment. Insulin sensitivity was estimated on the basis of glucose and insulin levels in the fasting state and after an oral glucose load. Participants in the DASH diet with aerobic exercise and caloric restriction condition lost weight (-8.7 kg [95% CI: -2.0 to -9.7 kg]) and exhibited a significant increase in aerobic capacity, whereas the DASH diet alone and UC participants maintained their weight (-0.3 kg [95% CI: -1.2 to 0.5 kg] and +0.9 kg [95% CI: 0.0 to 1.7 kg], respectively) and had no improvement in exercise capacity. DASH diet with aerobic exercise and caloric restriction demonstrated lower glucose levels after the oral glucose load, improved insulin sensitivity, and lower total cholesterol and triglycerides compared with both DASH diet alone and UC, as well as lower fasting glucose and low-density lipoprotein cholesterol compared with UC. DASH diet alone participants generally did not differ from UC in these measures. Combining the DASH diet with exercise and weight loss resulted in significant improvements in insulin sensitivity and lipids. Despite clinically significant reductions in blood pressure, the DASH diet alone, without caloric restriction or exercise, resulted in minimal improvements in insulin sensitivity or lipids.

The PROMIS Global Health Questionnaire Correlates With the QuickDASH in Patients With Upper Extremity Illness.

PubMed

Stoop, Nicky; Menendez, Mariano E; Mellema, Jos J; Ring, David

2018-01-01

The objective of this study is to evaluate the construct validity of the Patient-Reported Outcomes Measurement Information System (PROMIS) Global Health instrument by establishing its correlation to the Quick-Disabilities of the Arm, Shoulder and Hand (QuickDASH) questionnaire in patients with upper extremity illness. A cohort of 112 patients completed a sociodemographic survey and the PROMIS Global Health and QuickDASH questionnaires. Pearson correlation coefficients were used to evaluate the association of the QuickDASH with the PROMIS Global Health items and subscales. Six of the 10 PROMIS Global Health items were associated with the QuickDASH. The PROMIS Global Physical Health subscale showed moderate correlation with QuickDASH and the Mental Health subscale. There was no significant relationship between the PROMIS Global Mental Health subscale and QuickDASH. The consistent finding that general patient-reported outcomes correlate moderately with regional patient-reported outcomes suggests that a small number of relatively nonspecific patient-reported outcome measures might be used to assess a variety of illnesses. In our opinion, the blending of physical and mental health questions in the PROMIS Global Health makes this instrument less useful for research or patient care.

75 FR 8353 - Agency Information Collection Activities: Final Collection; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-24

... EXPORT[dash]IMPORT BANK OF THE U.S. [Public Notice 145] Agency Information Collection Activities: Final Collection; Comment Request AGENCY: Export-Import Bank of the U.S. ACTION: Submission for OMB... affects entities involved in the export of U.S. goods and services. Annual Number of Respondents: 84...

76 FR 54397 - Airworthiness Directives; Lycoming Engines (Type Certificate Previously Held by Textron Lycoming...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-01

...--Part Numbers (Including All Dash Numbers) of Known Affected HA-6 Model Carburetors 10-5219-XX 10-5224-XX 10-5230-XX 10-5235-XX 10-5253-XX 10-5255-XX 10-5283-XX 10-6001-XX 10-6019-XX 10-6030-XX Unsafe...

77 FR 16921 - Airworthiness Directives; Pratt & Whitney Division Turbofan Engines

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-23

... Airworthiness Directives; Pratt & Whitney Division Turbofan Engines AGENCY: Federal Aviation Administration (FAA..., PW4160, PW4460, PW4462, and PW4650 turbofan engines, including models with any dash number suffix. This... Compliance We estimate that this AD will affect 44 turbofan engines installed on airplanes of U.S. registry...

OHS Provides Easy Heart Health Tips at “DASH and Dine” Event | Poster

Cancer.gov

The lunchtime dash to a nearby eatery is a daily ritual at NCI at Frederick, but Occupational Health Services (OHS) recently showed employees a different, healthy kind of DASH—Dietary Approaches to Stop Hypertension. Employees who visited the educational lunch-hour event, called “DASH and Dine,” had the chance to speak with OHS staff about the benefits of the DASH Eating Plan, a diet developed by the National Heart, Lung, and Blood Institute (NHLBI).

Reliability, validity and responsiveness of the Arabic version of the Disability of Arm, Shoulder and Hand (DASH-Arabic).

PubMed

Alotaibi, Naser M; Aljadi, Sameera H; Alrowayeh, Hesham N

2016-12-01

To investigate the psychometric properties (reliability, validity and responsiveness) of the DASH-Arabic in a cohort of Arabic patients presenting with various upper extremity conditions. Participants were 139 patients with various upper extremity conditions, who completed the DASH-Arabic at the baseline, 2-5 days later and 30-36 days later. Participants completed demographic data forms, the SF-36 and VAS at baseline, and a Global Rating of Change scale at first and second follow-ups. Cronbach's alpha of the DASH-Arabic was 0.94. Test-retest reliability was excellent with an ICC of 0.97. The SEM was 3.50 and the MDC95 was 9.28. Construct validity of the DASH-Arabic with the SF-36 subscales and VAS scores ranged from r -0.32 to -0.57, all statistically significant (p < 0.001). The effect size (ES) for the DASH-Arabic was 1.39 and its standard response mean was 1.51. The area under the curve was 0.82 (95% CI = 0.72-0.92, p < 0.001). The optimally efficient cutoff for an improvement was found to be a difference of 15 DASH points. The DASH-Arabic is a reliable, valid and responsive upper extremity outcome measure for patients whose primary language is Arabic; it can be used to document patient status and outcomes and support evidence-based practice. Implications for Rehabilitation The DASH-Arabic demonstrated sound psychometric properties of reliability, validity and responsiveness. It is an effective patient status and outcome tool that will support evidence-based practice. This tool is recommended for evaluating upper extremity work-related injuries and tracking therapeutic outcomes.

Empirical evaluation of H.265/HEVC-based dynamic adaptive video streaming over HTTP (HEVC-DASH)

NASA Astrophysics Data System (ADS)

Irondi, Iheanyi; Wang, Qi; Grecos, Christos

2014-05-01

Real-time HTTP streaming has gained global popularity for delivering video content over Internet. In particular, the recent MPEG-DASH (Dynamic Adaptive Streaming over HTTP) standard enables on-demand, live, and adaptive Internet streaming in response to network bandwidth fluctuations. Meanwhile, emerging is the new-generation video coding standard, H.265/HEVC (High Efficiency Video Coding) promises to reduce the bandwidth requirement by 50% at the same video quality when compared with the current H.264/AVC standard. However, little existing work has addressed the integration of the DASH and HEVC standards, let alone empirical performance evaluation of such systems. This paper presents an experimental HEVC-DASH system, which is a pull-based adaptive streaming solution that delivers HEVC-coded video content through conventional HTTP servers where the client switches to its desired quality, resolution or bitrate based on the available network bandwidth. Previous studies in DASH have focused on H.264/AVC, whereas we present an empirical evaluation of the HEVC-DASH system by implementing a real-world test bed, which consists of an Apache HTTP Server with GPAC, an MP4Client (GPAC) with open HEVC-based DASH client and a NETEM box in the middle emulating different network conditions. We investigate and analyze the performance of HEVC-DASH by exploring the impact of various network conditions such as packet loss, bandwidth and delay on video quality. Furthermore, we compare the Intra and Random Access profiles of HEVC coding with the Intra profile of H.264/AVC when the correspondingly encoded video is streamed with DASH. Finally, we explore the correlation among the quality metrics and network conditions, and empirically establish under which conditions the different codecs can provide satisfactory performance.

Predictive value of the DASH tool for predicting return to work of injured workers with musculoskeletal disorders of the upper extremity.

PubMed

Armijo-Olivo, Susan; Woodhouse, Linda J; Steenstra, Ivan A; Gross, Douglas P

2016-12-01

To determine whether the Disabilities of the Arm, Shoulder, and Hand (DASH) tool added to the predictive ability of established prognostic factors, including patient demographic and clinical outcomes, to predict return to work (RTW) in injured workers with musculoskeletal (MSK) disorders of the upper extremity. A retrospective cohort study using a population-based database from the Workers' Compensation Board of Alberta (WCB-Alberta) that focused on claimants with upper extremity injuries was used. Besides the DASH, potential predictors included demographic, occupational, clinical and health usage variables. Outcome was receipt of compensation benefits after 3 months. To identify RTW predictors, a purposeful logistic modelling strategy was used. A series of receiver operating curve analyses were performed to determine which model provided the best discriminative ability. The sample included 3036 claimants with upper extremity injuries. The final model for predicting RTW included the total DASH score in addition to other established predictors. The area under the curve for this model was 0.77, which is interpreted as fair discrimination. This model was statistically significantly different than the model of established predictors alone (p<0.001). When comparing the DASH total score versus DASH item 23, a non-significant difference was obtained between the models (p=0.34). The DASH tool together with other established predictors significantly helped predict RTW after 3 months in participants with upper extremity MSK disorders. An appealing result for clinicians and busy researchers is that DASH item 23 has equal predictive ability to the total DASH score. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

One-Dimensional Nature of InAs/InP Quantum Dashes Revealed by Scanning Tunneling Spectroscopy.

PubMed

Papatryfonos, Konstantinos; Rodary, Guillemin; David, Christophe; Lelarge, François; Ramdane, Abderrahim; Girard, Jean-Christophe

2015-07-08

We report on low-temperature cross-sectional scanning tunneling microscopy and spectroscopy on InAs(P)/InGaAsP/InP(001) quantum dashes, embedded in a diode-laser structure. The laser active region consists of nine InAs(P) quantum dash layers separated by the InGaAsP quaternary alloy barriers. The effect of the p-i-n junction built-in potential on the band structure has been evidenced and quantified on large-scale tunneling spectroscopic measurements across the whole active region. By comparing the tunneling current onset channels, a consistent energy shift has been measured in successive quantum dash or barrier layers, either for the ground state energy of similar-sized quantum dashes or for the conduction band edge of the barriers, corresponding to the band-bending slope. The extracted values are in good quantitative agreement with the theoretical band structure calculations, demonstrating the high sensitivity of this spectroscopic measurement to probe the electronic structure of individual nanostructures, relative to local potential variations. Furthermore, by taking advantage of the potential gradient, we compared the local density of states over successive quantum dash layers. We observed that it does not vanish while increasing energy, for any of the investigated quantum dashes, in contrast to what would be expected for discrete level zero-dimensional (0D) structures. In order to acquire further proof and fully address the open question concerning the quantum dash dimensionality nature, we focused on individual quantum dashes obtaining high-energy-resolution measurements. The study of the local density of states clearly indicates a 1D quantum-wirelike nature for these nanostructures whose electronic squared wave functions were subsequently imaged by differential conductivity mapping.

A Multilevel Assessment of Barriers to Adoption of Dietary Approaches to Stop Hypertension (DASH) among African Americans of Low Socioeconomic Status

PubMed Central

Bertoni, Alain G.; Foy, Capri G.; Hunter, Jaimie C.; Quandt, Sara A.; Vitolins, Mara Z.; Whitt-Glover, Melicia C.

2013-01-01

Background We examined perceptions of Dietary Approaches to Stop Hypertension (DASH) and the food environment among African Americans (AA) with high blood pressure living in two low-income communities and objectively assessed local food outlets. Methods Focus groups were conducted with 30 AAs; participants discussed DASH and the availability of healthy foods in their community. Sessions were transcribed and themes identified. Fifty-four stores and 114 restaurants were assessed using the Nutrition Environment Measures Survey (NEMS). Results Common themes included poor availability, quality, and cost of healthy foods; tension between following DASH and feeding other family members; and lack of congruity between their preferred foods and DASH. Food outlets in majority AA census tracts had lower NEMS scores (stores: −11.7, p=.01, restaurants: −8.3, p=.001) compared with majority White areas. Conclusions Interventions promoting DASH among lower income AAs should reflect the food customs, economic concerns, and food available in communities. PMID:22080704

Measures of upper limb function for people with neck pain. A systematic review of measurement and practical properties.

PubMed

Alreni, Ahmad Salah Eldin; Harrop, Deborah; Lowe, Anna; Tanzila Potia; Kilner, Karen; McLean, Sionnadh Mairi

2017-06-01

There is a strong relationship between neck pain (NP) and upper limb disability (ULD). Optimal management of NP should incorporate upper limb rehabilitation and therefore include the use of an ULD measure in the assessment and management process. Clear guidance regarding the suitability of available measures does not exist. The aim of this study was to identify all available measures of ULD for populations with NP, critically evaluate their measurement properties and finally recommend a list of suitable measures. This two-phase systematic review is reported in accordance with the PRISMA statement. Phase one identified clearly reproducible measures of ULD for patients with NP. Phase two identified evidence of their measurement properties. In total, 11 papers evaluating the measurement properties of five instruments were included in this review. The instruments identified were the DASH questionnaire, the QuickDASH questionnaire, the NULI questionnaire, the SFA and the SAMP test. There was limited positive evidence of validity of the DASH, QuickDASH, NULI, SFA and SAMP. There was limited positive evidence of reliability of the NULI, SFA and SAMP. There was unknown evidence of responsiveness of the DASH and QuickDASH. Although all measures are supported by a limited amount of low quality evidence, the DASH, QuickDASH, NULI questionnaires, and the SAMP test are promising measures, but they require further robust evaluation. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

FMCSA’s advanced system testing utilizing a data acquisition system on the highways (FAST DASH) safety technology evaluation project #2 : technology brief.

DOT National Transportation Integrated Search

2016-11-01

The Federal Motor Carrier Safety Administration (FMCSA) established the FAST DASH program to perform efficient independent evaluations of promising safety technologies aimed at commercial vehicle operations. In this second FAST DASH safety technology...

Physical characterization of the cloned protease III gene from Escherichia coli K-12.

PubMed

Dykstra, C C; Kushner, S R

1985-09-01

Analysis of the cloned protease III gene (ptr) from Escherichia coli K-12 has demonstrated that in addition to the previously characterized 110,000-Mr protease III protein, a second 50,000-Mr polypeptide (p50) is derived from the amino-terminal end of the coding sequence. The p50 polypeptide is found predominantly in the periplasmic space along with protease III, but does not proteolytically degrade insulin, a substrate for protease III. p50 does not appear to originate from autolysis of the larger protein. Protease III is not essential for normal cell growth since deletion of the structural gene causes no observed alterations in the phenotypic properties of the bacteria. A 30-fold overproduction of protease III does not affect cell viability. A simple new purification method for protease III is described.

FMCSA’s advanced system testing utilizing a data acquisition system on the highways (FAST DASH) safety technology evaluation project #3 : novel convex mirrors : technology brief.

DOT National Transportation Integrated Search

2016-11-01

The Federal Motor Carrier Safety Administration (FMCSA) established the FAST DASH program to perform efficient independent evaluations of promising safety technologies aimed at commercial vehicle operations. In this third FAST DASH safety technology ...

Comparative responsiveness and minimal clinically important differences for idiopathic ulnar impaction syndrome.

PubMed

Kim, Jae Kwang; Park, Eun Soo

2013-05-01

Patient-reported questionnaires have been widely used to predict symptom severity and functional disability in musculoskeletal disease. Importantly, questionnaires can detect clinical changes in patients; however, this impact has not been determined for ulnar impaction syndrome. We asked (1) which of Patient-Rated Wrist Evaluation (PRWE), DASH, and other physical measures was more responsive to clinical improvements, and (2) what was the minimal clinically important difference for the PRWE and DASH after ulnar shortening osteotomy for idiopathic ulnar impaction syndrome. All patients who underwent ulnar shortening osteotomy between March 2008 and February 2011 for idiopathic ulnar impaction syndrome were enrolled in this study. All patients completed the PRWE and DASH questionnaires, and all were evaluated for grip strength and wrist ROM, preoperatively and 12 months postoperatively. We compared the effect sizes observed by each of these instruments. Effect size is calculated by dividing the mean change in a score of each instrument during a specified interval by the standard deviation of the baseline score. In addition, patient-perceived overall improvement was used as the anchor to determine the minimal clinically important differences on the PRWE and DASH 12 months after surgery. The average score of each item except for wrist flexion and supination improved after surgery. The PRWE was more sensitive than the DASH or than physical measurements in detecting clinical changes. The effect sizes and standardized response means of the outcome measures were as follows: PRWE (1.51, 1.64), DASH (1.12, 1.24), grip strength (0.59, 0.68), wrist pronation (0.33, 0.41), and wrist extension (0.28, 0.36). Patient-perceived overall improvement and score changes of the PRWE and DASH correlated significantly. Minimal clinically important differences were 17 points (of a possible 100) for the PRWE and 13.5 for the DASH (also of 100), and minimal detectable changes were 7.7 points for the PRWE and 9.3 points for the DASH. Although the PRWE and DASH were highly sensitive to clinical changes, the PRWE was more sensitive in terms of detecting clinical changes after ulnar shortening osteotomy for idiopathic ulnar impaction syndrome. A minimal change of 17 PRWE points or 13.5 DASH points was necessary to achieve a benefit that patients perceived as clinically important. The minimal clinically important differences using these instruments were higher than the values produced by measurement errors.

Formation of unexpected silicon- and disiloxane-bridged multiferrocenyl derivatives bearing Si-O-CH[double bond, length as m-dash]CH2 and Si-(CH2)2C(CH3)3 substituents via cleavage of tetrahydrofuran and trapping of its ring fragments.

PubMed

Bruña, Sonia; González-Vadillo, Ana Mª; Ferrández, Marta; Perles, Josefina; Montero-Campillo, M Merced; Mó, Otilia; Cuadrado, Isabel

2017-09-12

The formation of a family of silicon- and siloxane-bridged multiferrocenyl derivatives carrying different functional groups attached to silicon, including Fc 2 (CH 3 ) 3 C(CH 2 ) 2 SiCH[double bond, length as m-dash]CH 2 (5), Fc 2 (CH 2 [double bond, length as m-dash]CH-O)SiCH[double bond, length as m-dash]CH 2 (6), Fc 2 (OH)SiCH[double bond, length as m-dash]CH 2 (7), Fc 2 (CH 2 [double bond, length as m-dash]CH-O)Si-O-Si(O-CH[double bond, length as m-dash]CH 2 )Fc 2 (8) and Fc 2 (CH 2 [double bond, length as m-dash]CH-O)Si-O-SiFc 3 (9) is described. Silyl vinyl ether molecules 6, 8 and 9 and the heteroleptic vinylsilane 5 resulted from the competing metathesis reaction of lithioferrocene (FcLi), CH 2 [double bond, length as m-dash]CH-OLi or (CH 3 ) 3 C(CH 2 ) 2 Li with the corresponding multifunctional chlorosilane, Cl 3 SiCH[double bond, length as m-dash]CH 2 or Cl 3 Si-O-SiCl 3 . The last two organolithium species have been likely formed in situ by fragmentation of the tetrahydrofuran solvent. Diferrocenylvinyloxyvinylsilane 6 is noteworthy since it represents a rare example of a redox-active silyl mononomer in which two different C[double bond, length as m-dash]C polymerisable groups are directly connected to silicon. The molecular structures of the silicon-containing multiferrocenyl species 5, 6, 8 and 9 have been investigated by single-crystal X-ray diffraction studies, demonstrating the capture and storage processes of two ring fragments resulting from the cleavage of cyclic THF in redox-active and stable crystalline organometallic compounds. From electrochemical studies we found that by changing the anion of the supporting electrolyte from [PF 6 ] - to [B(C 6 F 5 ) 4 ] - , the redox behaviour of tetrametallic disiloxane 8 can be switched from a poorly resolved multistep redox process to four consecutive well-separated one-electron oxidations, corresponding to the sequential oxidation of the four ferrocenyl moieties.

A P-H functionalized Al/P-based frustrated Lewis pair - hydrophosphination of nitriles, ring opening with cyclopropenones and evidence of P[double bond, length as m-dash]C double bond formation.

PubMed

Keweloh, Lukas; Aders, Niklas; Hepp, Alexander; Pleschka, Damian; Würthwein, Ernst-Ulrich; Uhl, Werner

2018-06-12

Hydroalumination of R-P(H)-C[triple bond, length as m-dash]C-tBu with bulky H-Al[CH(SiMe3)2]2 afforded the new P-H functionalized Al/P-based frustrated Lewis pair R-P(H)-C[[double bond, length as m-dash]C(H)-tBu]-AlR2 [R = CH(SiMe3)2; FLP 7]. A weak adduct of 7 with benzonitrile (8) was detected by NMR spectroscopy, but could not be isolated. tert-Butyl isocyanide afforded a similar, but isolable adduct (9), in which the isocyanide C atom was coordinated to aluminium. The unique reactivity of 7 became evident from its reactions with the heteroatom substituted nitriles PhO-C[triple bond, length as m-dash]N, PhCH2S-C[triple bond, length as m-dash]N and H8C4N-C[triple bond, length as m-dash]N. Hydrophosphination of the C[triple bond, length as m-dash]N triple bonds afforded imines at room temperature which were coordinated to the FLP by Al-N and P-C bonds to yield AlCPCN heterocycles (10 to 12). These processes depend on substrate activation by the FLP. Diphenylcyclopropenone and its sulphur derivative reacted with 7 by addition of the P-H bond to a C-C bond of the strained C3 ring and ring opening to afford the fragment (Z)-Ph-C(H)[double bond, length as m-dash]C(Ph)-C-X-Al (X = O, S). The C-O or C-S groups were coordinated to the FLP to yield AlCPCX heterocycles (13 and 14). The thiocarbonyl derived compound 14 contains an internally stabilized phosphenium cation with a localized P[double bond, length as m-dash]C bond, a trigonal planar coordinated P atom and a short P[double bond, length as m-dash]C distance (168.9 pm). Insight into formation mechanisms, the structural and energetic properties of FLP 7 and compounds 13 and 14 was gained by quantum chemical DFT calculations.

Relation between the Disability of the Arm, Shoulder and Hand Score and Muscle Strength in Post-Cardiac Surgery Patients.

PubMed

Izawa, Kazuhiro P; Kasahara, Yusuke; Hiraki, Koji; Hirano, Yasuyuki; Watanabe, Satoshi

2017-11-27

Background: The Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire is a valid and reliable patient-reported outcome measure. DASH can be assessed by self-reported upper extremity disability and symptoms. We aimed to examine the relationship between the physiological outcome of muscle strength and the DASH score after cardiac surgery. Methods: This cross-sectional study assessed 50 consecutive cardiac patients that were undergoing cardiac surgery. Physiological outcomes of handgrip strength and knee extensor muscle strength and the DASH score were measured at one month after cardiac surgery and were assessed. Results were analyzed using Spearman correlation coefficients. Results: The final analysis comprised 43 patients (men: 32, women: 11; age: 62.1 ± 9.1 years; body mass index: 22.1 ± 4.7 kg/m²; left ventricular ejection fraction: 53.5 ± 13.7%). Respective handgrip strength, knee extensor muscle strength, and DASH score were 27.4 ± 8.3 kgf, 1.6 ± 0.4 Nm/kg, and 13.3 ± 12.3, respectively. The DASH score correlated negatively with handgrip strength ( r = -0.38, p = 0.01) and with knee extensor muscle strength ( r = -0.32, p = 0.04). Conclusion: Physiological outcomes of both handgrip strength and knee extensor muscle strength correlated negatively with the DASH score. The DASH score appears to be a valuable tool with which to assess cardiac patients with poor physiological outcomes, particularly handgrip strength as a measure of upper extremity function, which is probably easier to follow over time than lower extremity function after patients complete cardiac rehabilitation.

Disability in the upper extremity and quality of life in hand-arm vibration syndrome.

PubMed

Poole, Kerry; Mason, Howard

2005-11-30

To investigate whether hand-arm vibration syndrome (HAVS) leads to disability in the upper extremity or deficit in quality of life (QoL) using validated questionnaire tools, and to establish whether these effects are related to the Stockholm Workshop Staging (SWS). This was a postal cross-sectional questionnaire study with a 50% response rate. Four hundred and forty-four males, who had been diagnosed and staged according to the SWS were sent the Disability in the Arm, Shoulder and Hand (DASH) and the SF-36v2 QoL questionnaires. HAVS cases had significantly greater DASH disability scores and reduced QoL physical and mental component scores compared to published normal values. Those HAVS cases with a presumptive diagnosis of Carpal Tunnel Syndrome(CTS) had even higher disability scores. There was a clear, linear relationship between both the DASH disability score and the physical component of the QoL and sensorineural SWS, but not with the vascular SWS. HAVS has a significant effect on an individual's perceived ability to perform everyday tasks involving the upper extremity, and their quality of life. Physical capability may be further compromised in those individuals who have a presumptive diagnosis of CTS. These findings may have important implications regarding management of the affected worker.

Reliability, Validity, and Responsiveness of the QuickDASH in Patients With Upper Limb Amputation.

PubMed

Resnik, Linda; Borgia, Matthew

2015-09-01

To examine the internal consistency, test-retest reliability, validity, and responsiveness of the shortened version of the Disabilities of the Arm, Shoulder and Hand (QuickDASH) questionnaire in persons with upper limb amputation. Cross-sectional and longitudinal. Three sites participating in the U.S. Department of Veterans Affairs Home Study of the DEKA Arm. A convenience sample of upper limb amputees (N=44). Training with a multifunction upper limb prosthesis. Multiple outcome measures including the QuickDASH were administered twice within 1 week, and for a subset of 20 persons, after completion of in-laboratory training with the DEKA Arm. Scale alphas and intraclass correlation coefficient type 3,1 (ICC3,1) were used to examine reliability. Minimum detectable change (MDC) scores were calculated. Analyses of variance, comparing QuickDASH scores by the amount of prosthetic use and amputation level, were used for known-group validity analyses with alpha set at .05. Pairwise correlations between QuickDASH and other measures were used to examine concurrent validity. Responsiveness was measured by effect size (ES) and standardized response mean (SRM). QuickDASH alpha was .83, and ICC was .87 (95% confidence interval, .77-.93). MDC at the 95% confidence level (MDC95%) was 17.4. Full- or part-time prosthesis users had better QuickDASH scores compared with nonprosthesis users (P=.021), as did those with more distal amputations at both baseline (P=.042) and with the DEKA Arm (P=.024). The QuickDASH was correlated with concurrent measures of activity limitation as expected. The ES and SRM after training with the DEKA Arm were 0.6. This study provides evidence of reliability and validity of the QuickDASH in persons with upper limb amputation. Results provide preliminary evidence of responsiveness to prosthetic device type/training. Further research with a larger sample is needed to confirm results. Copyright © 2015 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.

Lazarus and Group Psychotherapy: AIDS in the Era of Protease Inhibitors

ERIC Educational Resources Information Center

Gushue, George V.; Brazaitis, Sarah J.

2003-01-01

A new class of medications, protease inhibitors, has dramatically improved the health of many people with Human Immunodeficiency Virus (HIV) and Acquired Immune Deficiency Syndrome (AIDS). This development has had a major impact on the lives of those affected by HIV/AIDS. This article considers how a group is affected by the larger systems of…

Zebra chip disease decreases tuber (Solanum tuberosum L.) protein content by attenuating protease inhibitor levels and increasing protease activities.

PubMed

Kumar, G N Mohan; Knowles, Lisa O; Knowles, N Richard

2015-11-01

Zebra chip disease of potato decreases protease inhibitor levels resulting in enhanced serine-type protease activity, decreased protein content and altered protein profiles of fully mature tubers. Zebra-chip (ZC), caused by Candidatus Liberibacter solanacearum (CLso), is a relatively new disease of potato that negatively affects growth, yield, propagation potential, and fresh and process qualities of tubers. Diseased plants produce tubers with characteristic brown discoloration of vascular tissue accompanied by elevated levels of free amino acids and reducing sugars. Here we demonstrate that ZC disease induces selective protein catabolism in tubers through modulating protease inhibitor levels. Soluble protein content of tubers from CLso-infected plants was 33% lower than from non-infected plants and electrophoretic analyses revealed substantial reductions in major tuber proteins. Patatin (~40 kDa) and ser-, asp- (22 kDa) and cys-type (85 kDa) protease inhibitors were either absent or greatly reduced in ZC-afflicted tubers. In contrast to healthy (non-infected) tubers, the proteolytic activity in CLso infected tubers was high and the ability of extracts from infected tubers to inhibit trypsin (ser-type) and papain (cys-type) proteases greatly attenuated. Moreover, extracts from CLso-infected tubers rapidly catabolized proteins purified from healthy tubers (40 kDa patatin, 22 kDa protease inhibitors, 85 kDa potato multicystatin) when subjected to proteolysis individually. In contrast, crude extracts from non-infected tubers effectively inhibited the proteolytic activity from ZC-afflicted tubers. These results suggest that the altered protein profile of ZC afflicted tubers is largely due to loss of ser- and cys-type protease inhibitors. Further analysis revealed a novel PMSF-sensitive (ser) protease (ca. 80-120 kDa) in CLso infected tubers. PMSF abolished the proteolytic activities responsible for degrading patatin, the 22 kDa protease inhibitor(s) and potato multicystatin by CLso infected tubers. The disease-induced loss of patatin and protease inhibitors therefore appears to be modulated by ser-type protease(s). The selective catabolism of proteins in ZC-afflicted tubers undoubtedly affects downstream aspects of carbohydrate and amino acid metabolism, which is ultimately reflected by the accumulation of reducing sugars, free amino acids and reduced sprouting capacity.

Evaluating the safety and efficacy of sodium-restricted/Dietary Approaches to Stop Hypertension diet after acute decompensated heart failure hospitalization: design and rationale for the Geriatric OUt of hospital Randomized MEal Trial in Heart Failure (GOURMET-HF).

PubMed

Wessler, Jeffrey D; Maurer, Mathew S; Hummel, Scott L

2015-03-01

Heart failure (HF) is a major public health problem affecting predominantly older adults. Nonadherence to diet remains a significant contributor to acute decompensated HF (ADHF). The sodium-restricted Dietary Approaches to Stop Hypertension (DASH/SRD) eating plan reduces cardiovascular dysfunction that can lead to ADHF and is consistent with current HF guidelines. We propose that an intervention that promotes adherence to the DASH/SRD by home-delivering meals will be safe and improve health-related quality of life (QOL) in older adults after hospitalization for ADHF. This is a 3-center, randomized, single-blind, controlled trial of 12-week duration designed to determine the safety and efficacy of home-delivered DASH/SRD-compliant meals in older adults after discharge from ADHF hospitalization. Sixty-six subjects will be randomized in a 1:1 stratified fashion by gender and left ventricular ejection fraction (<50% vs ≥50%). Study subjects will receive either preprepared, home-delivered DASH/SRD-compliant meals or usual dietary advice for 4weeks after hospital discharge. Investigators will be blinded to group assignment, food diaries, and urinary electrolyte measurements until study completion. The primary efficacy end point is the change in the Kansas City Cardiomyopathy Questionnaire summary scores for health-related QOL from study enrollment to 4weeks postdischarge. Safety evaluation will focus on hypotension, renal insufficiency, and hyperkalemia. Exploratory end points include echocardiography, noninvasive vascular testing, markers of oxidative stress, and salt taste sensitivity. This randomized controlled trial will test the efficacy, feasibility, and safety of 4weeks of DASH/SRD after ADHF hospitalization. By testing a novel dietary intervention supported by multiple levels of evidence including preliminary data in outpatients with stable HF, we will address a critical evidence gap in the care of older patients with ADHF. If effective and safe, this intervention could be scaled to assess effects on readmission and healthcare costs in older adults after ADHF. Published by Elsevier Inc.

Analysis of factors related to arm weakness in patients with breast cancer-related lymphedema.

PubMed

Lee, Daegu; Hwang, Ji Hye; Chu, Inho; Chang, Hyun Ju; Shim, Young Hun; Kim, Jung Hyun

2015-08-01

The aim of this study was to evaluate the ratio of significant weakness in the affected arm of breast cancer-related lymphedema patients to their unaffected side. Another purpose was to identify factors related to arm weakness and physical function in patients with breast cancer-related lymphedema. Consecutive patients (n = 80) attended a single evaluation session following their outpatient lymphedema clinic visit. Possible independent factors (i.e., lymphedema, pain, psychological, educational, and behavioral) were evaluated. Handgrip strength was used to assess upper extremity muscle strength and the disabilities of arm, shoulder, and hand (DASH) questionnaire was used to assess upper extremity physical function. Multivariate logistic regression was performed using factors that had significant differences between the handgrip weakness and non-weakness groups. Out of the 80 patients with breast cancer-related lymphedema, 29 patients (36.3 %) had significant weakness in the affected arm. Weakness of the arm with lymphedema was not related to lymphedema itself, but was related to the fear of using the affected limb (odds ratio = 1.76, 95 % confidence interval = 1.30-2.37). Fears of using the affected limb and depression significantly contributed to the variance in DASH scores. Appropriate physical and psychological interventions, including providing accurate information and reassurance of physical activity safety, are necessary to prevent arm weakness and physical dysfunction in patients with breast cancer-related lymphedema.

Dietary Approaches to Stop Hypertension Diet, Weight Status, and Blood Pressure among Children and Adolescents: National Health and Nutrition Examination Surveys 2003-2012.

PubMed

Cohen, Juliana F W; Lehnerd, Megan E; Houser, Robert F; Rimm, Eric B

2017-09-01

The Dietary Approaches to Stop Hypertension (DASH) diet is associated with lower blood pressure and reduced risk of cardiovascular disease among adults, but little is known about accordance with this dietary pattern or health benefits among children and adolescents. The objectives were to evaluate accordance with the DASH diet, differences over time, and the association with health attributes among a nationally representative sample of US children and adolescents. Cross-sectional data from the 2003-2012 National Health and Nutrition Examination Surveys (NHANES) were analyzed. Data from 9,793 individuals aged 8 to 18 years were examined. DASH accordance was estimated based on nine nutrient targets: total fat, saturated fat, protein, cholesterol, fiber, calcium, magnesium, potassium, and sodium; possible score range is 0 to 9. Accordance with the DASH diet across time was examined comparing the 2011-2012 to 2003-2004 NHANES surveys. The association between DASH score and weight status was examined using multinomial logistic regression, and the associations with waist circumference, systolic blood pressure, and diastolic blood pressure were examined using multivariable linear regression. Accordance with the DASH diet was low across the age groups, with a range of mean DASH scores from 1.48 to 2.14. There were no significant changes across time. DASH score was inversely associated with systolic blood pressure (mm Hg) among 14- to 18-year-olds (β=-.46; 95% CI -.83 to -.09) among the larger sample of participants who completed at least one dietary recall, but no significant differences were seen in other age categories. In the subsample of participants with both dietary recalls, a significant inverse association was seen between DASH score and systolic blood pressure for 11- to 13-year-olds (β=-.57; 95% CI -1.02 to -.12). There were no significant associations between this dietary pattern and weight status, waist circumference, or diastolic blood pressure. Few US children and adolescents have diets that are in alignment with the DASH diet. Future research should explore strategies to encourage fruit, vegetable, and whole-grain consumption, as well as sodium reductions to help meet DASH nutrient targets in children and adolescents, as well as examine the potential benefits of this eating pattern on health in this population group. Copyright © 2017 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

A Look Inside HIV Resistance through Retroviral Protease Interaction Maps

PubMed Central

Kontijevskis, Aleksejs; Prusis, Peteris; Petrovska, Ramona; Yahorava, Sviatlana; Mutulis, Felikss; Mutule, Ilze; Komorowski, Jan; Wikberg, Jarl E. S

2007-01-01

Retroviruses affect a large number of species, from fish and birds to mammals and humans, with global socioeconomic negative impacts. Here the authors report and experimentally validate a novel approach for the analysis of the molecular networks that are involved in the recognition of substrates by retroviral proteases. Using multivariate analysis of the sequence-based physiochemical descriptions of 61 retroviral proteases comprising wild-type proteases, natural mutants, and drug-resistant forms of proteases from nine different viral species in relation to their ability to cleave 299 substrates, the authors mapped the physicochemical properties and cross-dependencies of the amino acids of the proteases and their substrates, which revealed a complex molecular interaction network of substrate recognition and cleavage. The approach allowed a detailed analysis of the molecular–chemical mechanisms involved in substrate cleavage by retroviral proteases. PMID:17352531

Subjective evaluation of H.265/HEVC based dynamic adaptive video streaming over HTTP (HEVC-DASH)

NASA Astrophysics Data System (ADS)

Irondi, Iheanyi; Wang, Qi; Grecos, Christos

2015-02-01

The Dynamic Adaptive Streaming over HTTP (DASH) standard is becoming increasingly popular for real-time adaptive HTTP streaming of internet video in response to unstable network conditions. Integration of DASH streaming techniques with the new H.265/HEVC video coding standard is a promising area of research. The performance of HEVC-DASH systems has been previously evaluated by a few researchers using objective metrics, however subjective evaluation would provide a better measure of the user's Quality of Experience (QoE) and overall performance of the system. This paper presents a subjective evaluation of an HEVC-DASH system implemented in a hardware testbed. Previous studies in this area have focused on using the current H.264/AVC (Advanced Video Coding) or H.264/SVC (Scalable Video Coding) codecs and moreover, there has been no established standard test procedure for the subjective evaluation of DASH adaptive streaming. In this paper, we define a test plan for HEVC-DASH with a carefully justified data set employing longer video sequences that would be sufficient to demonstrate the bitrate switching operations in response to various network condition patterns. We evaluate the end user's real-time QoE online by investigating the perceived impact of delay, different packet loss rates, fluctuating bandwidth, and the perceived quality of using different DASH video stream segment sizes on a video streaming session using different video sequences. The Mean Opinion Score (MOS) results give an insight into the performance of the system and expectation of the users. The results from this study show the impact of different network impairments and different video segments on users' QoE and further analysis and study may help in optimizing system performance.

Relation between the Disability of the Arm, Shoulder and Hand Score and Muscle Strength in Post-Cardiac Surgery Patients

PubMed Central

Kasahara, Yusuke; Hiraki, Koji; Hirano, Yasuyuki; Watanabe, Satoshi

2017-01-01

Background: The Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire is a valid and reliable patient-reported outcome measure. DASH can be assessed by self-reported upper extremity disability and symptoms. We aimed to examine the relationship between the physiological outcome of muscle strength and the DASH score after cardiac surgery. Methods: This cross-sectional study assessed 50 consecutive cardiac patients that were undergoing cardiac surgery. Physiological outcomes of handgrip strength and knee extensor muscle strength and the DASH score were measured at one month after cardiac surgery and were assessed. Results were analyzed using Spearman correlation coefficients. Results: The final analysis comprised 43 patients (men: 32, women: 11; age: 62.1 ± 9.1 years; body mass index: 22.1 ± 4.7 kg/m2; left ventricular ejection fraction: 53.5 ± 13.7%). Respective handgrip strength, knee extensor muscle strength, and DASH score were 27.4 ± 8.3 kgf, 1.6 ± 0.4 Nm/kg, and 13.3 ± 12.3, respectively. The DASH score correlated negatively with handgrip strength (r = −0.38, p = 0.01) and with knee extensor muscle strength (r = −0.32, p = 0.04). Conclusion: Physiological outcomes of both handgrip strength and knee extensor muscle strength correlated negatively with the DASH score. The DASH score appears to be a valuable tool with which to assess cardiac patients with poor physiological outcomes, particularly handgrip strength as a measure of upper extremity function, which is probably easier to follow over time than lower extremity function after patients complete cardiac rehabilitation. PMID:29186880

Role of tissue factor and protease-activated receptors in a mouse model of endotoxemia.

PubMed

Pawlinski, Rafal; Pedersen, Brian; Schabbauer, Gernot; Tencati, Michael; Holscher, Todd; Boisvert, William; Andrade-Gordon, Patricia; Frank, Rolf Dario; Mackman, Nigel

2004-02-15

Sepsis is associated with a systemic activation of coagulation and an excessive inflammatory response. Anticoagulants have been shown to inhibit both coagulation and inflammation in sepsis. In this study, we used both genetic and pharmacologic approaches to analyze the role of tissue factor and protease-activated receptors in coagulation and inflammation in a mouse endotoxemia model. We used mice expressing low levels of the procoagulant molecule, tissue factor (TF), to analyze the effects of TF deficiency either in all tissues or selectively in hematopoietic cells. Low TF mice had reduced coagulation, inflammation, and mortality compared with control mice. Similarly, a deficiency of TF expression by hematopoietic cells reduced lipopolysaccharide (LPS)-induced coagulation, inflammation, and mortality. Inhibition of the down-stream coagulation protease, thrombin, reduced fibrin deposition and prolonged survival without affecting inflammation. Deficiency of either protease activated receptor-1 (PAR-1) or protease activated receptor-2 (PAR-2) alone did not affect inflammation or survival. However, a combination of thrombin inhibition and PAR-2 deficiency reduced inflammation and mortality. These data demonstrate that hematopoietic cells are the major pathologic site of TF expression during endotoxemia and suggest that multiple protease-activated receptors mediate crosstalk between coagulation and inflammation.

Mid-term functional outcome after the internal fixation of distal radius fractures

PubMed Central

2012-01-01

Background Distal radius fracture is a common injury with a variety of operative and non-operative management options. There remains debate as to the optimal treatment for a given patient and fracture. Despite the popularity of volar locking plate fixation, there are few large cohort or long term follow up studies to justify this modality. Our aim was to report the functional outcome of a large number of patients at a significant follow up time after fixation of their distal radius with a volar locking plate. Methods 180 patients with 183 fractures and a mean age of 62.4 years were followed up retrospectively at a mean of 30 months (Standard deviation = 10.4). Functional assessment was performed using the Disabilities of the Arm, Shoulder and Hand (DASH) and modified MAYO wrist scores. Statistical analysis was performed to identify possible variables affecting outcome and radiographs were assessed to determine time to fracture union. Results The median DASH score was 2.3 and median MAYO score was 90 for the whole group. Overall, 133 patients (74%) had a good or excellent DASH and MAYO score. Statistical analysis showed that no specific variable including gender, age, fracture type, post-operative immobilisation or surgeon grade significantly affected outcome. Complications occurred in 27 patients (15%) and in 11 patients were major (6%). Conclusion This single centre large population series demonstrates good to excellent results in the majority of patients after volar locking plate fixation of the distal radius, with complication rates comparable to other non-operative and operative treatment modalities. On this basis we recommend this mode of fixation for distal radius fractures requiting operative intervention. PMID:22280557

Mid-term functional outcome after the internal fixation of distal radius fractures.

PubMed

Phadnis, Joideep; Trompeter, Alex; Gallagher, Kieran; Bradshaw, Lucy; Elliott, David S; Newman, Kevin J

2012-01-26

Distal radius fracture is a common injury with a variety of operative and non-operative management options. There remains debate as to the optimal treatment for a given patient and fracture. Despite the popularity of volar locking plate fixation, there are few large cohort or long term follow up studies to justify this modality. Our aim was to report the functional outcome of a large number of patients at a significant follow up time after fixation of their distal radius with a volar locking plate. 180 patients with 183 fractures and a mean age of 62.4 years were followed up retrospectively at a mean of 30 months (Standard deviation=10.4). Functional assessment was performed using the Disabilities of the Arm, Shoulder and Hand (DASH) and modified MAYO wrist scores. Statistical analysis was performed to identify possible variables affecting outcome and radiographs were assessed to determine time to fracture union. The median DASH score was 2.3 and median MAYO score was 90 for the whole group. Overall, 133 patients (74%) had a good or excellent DASH and MAYO score. Statistical analysis showed that no specific variable including gender, age, fracture type, post-operative immobilisation or surgeon grade significantly affected outcome. Complications occurred in 27 patients (15%) and in 11 patients were major (6%). This single centre large population series demonstrates good to excellent results in the majority of patients after volar locking plate fixation of the distal radius, with complication rates comparable to other non-operative and operative treatment modalities. On this basis we recommend this mode of fixation for distal radius fractures requiting operative intervention.

The effect of redox-active cyanomanganese(I) ligands on intramolecular electron transfer to, and alkyne alignment in, M(CO)(RC[triple bond, length as m-dash]CR)Tp' (M = Mo or W) units.

PubMed

Adams, Christopher J; Connelly, Neil G; Onganusorn, Sriwipha

2009-04-28

The complexes [(eta-C(5)Me(5))(ON)LMn(micro-CN)M(CO)(RC[triple bond, length as m-dash]CR)Tp'](+) (L = CNXyl, M = Mo; L = CNBu(t), M = Mo or W, R = Ph or Me) and trans- or cis-[(dppm){(EtO)(3)P}(OC)(2)Mn(micro-CN)M(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](+), and their linkage isomers [(eta-C(5)Me(5))(ON)LMn(micro-NC)M(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](+) and trans- or cis-[(dppm){(EtO)(3)P}(OC)(2)Mn(micro-NC)M(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](+), undergo two one-electron oxidations. The complexes [(eta-C(5)Me(5))(ON)LMn(micro-XY)M(CO)(RC[triple bond, length as m-dash]CR)Tp'](+) (XY = CN or NC) are oxidised first at the N-bound metal centre and then at the C-bound centre. For [(dppm){(EtO)(3)P}(OC)(2)Mn(micro-XY)M(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](+), the trans isomers are first oxidised at manganese whereas the cis isomers are first oxidised at M. Thus, the order of one-electron oxidation of the two series of binuclear monocations is influenced by linkage isomerisation of the cyanide bridge and cis-trans isomerisation of the Mn(CO)(2) group. IR spectroscopic changes on reaction of Ag(+) with [(eta-C(5)Me(5))(ON)(Bu(t)NC)Mn(micro-CN)W(CO)(MeC[triple bond, length as m-dash]CMe)Tp'](+) are consistent with one-electron at the N-bound tungsten centre. Likewise, trans-[(dppm){(EtO)(3)P}(OC)(2)Mn(micro-NC)M(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](+) (M = Mo or W) give the stable dications [(dppm){(EtO)(3)P}(OC)(2)Mn(micro-NC)M(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](2+). Significantly longer Mn-P bond distances in trans-[(dppm){(EtO)(3)P}(OC)(2)Mn(micro-NC)Mo(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](2+) than in trans-[(dppm){(EtO)(3)P}(OC)(2)Mn(micro-NC)Mo(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](+) are consistent with one-electron oxidation first at Mn(I); the alignment of the (CN)Mn(CO)(2){P(OEt)(3)}(dppm) fragment relative to the alkyne in trans-[(dppm){(EtO)(3)P}(OC)(2)Mn(micro-NC)Mo(CO)(PhC[triple bond, length as m-dash]CPh)Tp'](+) suggests it acts as a pi-acceptor, in contrast to related species such as trans-(NC)Mn(CO)(2){P(OEt)(3)}(dppm) and (NC)Mn(NO){P(OPh)(3)}(pi-C(5)H(4)Me) which behave as simple N-donors.

An unusual alkylidyne homologation.

PubMed

Han, Yong-Shen; Hill, Anthony F; Kong, Richard Y

2018-02-27

The reaction of [W([triple bond, length as m-dash]CH)Br(CO) 2 (dcpe)] (dcpe = 1,2-bis(dicyclohexylphosphino)ethane) with t BuLi and SiCl 4 affords the trichlorosilyl ligated neopentylidyne complex [W([triple bond, length as m-dash]C t Bu)(SiCl 3 )(CO) 2 (dcpe)]. This slowly reacts with H 2 O to afford [W([triple bond, length as m-dash]CCH 2 t Bu)Cl 3 (dcpe)] and ultimately H 2 C[double bond, length as m-dash]CH t Bu via an unprecedented alkylidyne homologation in which coordinated CO is the source of the additional carbon atom with potential relevance to the Fischer-Tropsch process.

Comparison of the Properties of the Handwriting Speed Test (HST) and Detailed Assessment of Speed of Handwriting (DASH): An Exploratory Study.

PubMed

Francis, Anna; Wallen, Margaret; Bundy, Anita

2017-05-01

Handwriting speed is an important component of students' ability to adequately express their ideas, knowledge and creativity in a timely and effective manner. Psychometric properties of the Handwriting Speed Test (HST) and Detailed Assessment of Speed of Handwriting (DASH) and accuracy of the norms for identifying current Australian students with handwriting speed difficulties were examined. An exploratory, cross-sectional study was conducted involving students, with and without handwriting difficulties, in Years 3-12 (mean age: 12.0 yrs, SD = 3.0 yrs; range = 7 to 18 yrs) in New South Wales (NSW; Australia). Participants were recruited through occupational therapists and schools. Students completed the HST and all DASH subtests. Thirty-two students with, and 139 students without, handwriting difficulties participated. Intra-rater and inter-rater reliability were found to be excellent; sensitivity was low and specificity high for the HST and DASH. No significant differences were found between test scores and normative data for students without handwriting difficulties (year/age groups with n > 10). The HST and DASH are reliable assessments of handwriting speed. Further research is required into discriminant validity of the HST and DASH and need for updated norms.

Construct validity, test-retest reliability and internal consistency of the Thai version of the disabilities of the arm, shoulder and hand questionnaire (DASH-TH) in patients with carpal tunnel syndrome.

PubMed

Buntragulpoontawee, Montana; Phutrit, Suphatha; Tongprasert, Siam; Wongpakaran, Tinakon; Khunachiva, Jeeranan

2018-03-27

This study evaluated additional psychometric properties of the Thai version of the disabilities of the arm, shoulder and hand questionnaire (DASH-TH) which included, test-retest reliability, construct validity, internal consistency of in patients with carpal tunnel syndrome. As for determining construct validity, the Thai EuroQOL questionnaire (EQ-5D-5L) was also administered in order to examine convergent and divergent validity. Fifty patients completed both questionnaires. The DASH-TH showed excellent test-retest reliability (intraclass correlation coefficient = 0.811) and internal consistency (Cronbach's alpha = 0.911). The exploratory factor analysis yielded a six-factor solution while the confirmatory factor analysis denoted that the hypothesized model adequately fit the data with a comparative fit index of 0.967 and a Tucker-Lewis index of 0.964. The related subscales between the DASH-TH and the Thai EQ-5D-5L were significantly correlated, indicating the DASH-TH's convergent and discriminant validity. The DASH-TH demonstrated good reliability, internal consistency construct validity, and multidimensionality, in assessing the upper extremity function in carpal tunnel syndrome patients.

Cysteine Protease Inhibitors as Chemotherapy: Lessons from a Parasite Target

NASA Astrophysics Data System (ADS)

Selzer, Paul M.; Pingel, Sabine; Hsieh, Ivy; Ugele, Bernhard; Chan, Victor J.; Engel, Juan C.; Bogyo, Matthew; Russell, David G.; Sakanari, Judy A.; McKerrow, James H.

1999-09-01

Papain family cysteine proteases are key factors in the pathogenesis of cancer invasion, arthritis, osteoporosis, and microbial infections. Targeting this enzyme family is therefore one strategy in the development of new chemotherapy for a number of diseases. Little is known, however, about the efficacy, selectivity, and safety of cysteine protease inhibitors in cell culture or in vivo. We now report that specific cysteine protease inhibitors kill Leishmania parasites in vitro, at concentrations that do not overtly affect mammalian host cells. Inhibition of Leishmania cysteine protease activity was accompanied by defects in the parasite's lysosome/endosome compartment resembling those seen in lysosomal storage diseases. Colocalization of anti-protease antibodies with biotinylated surface proteins and accumulation of undigested debris and protease in the flagellar pocket of treated parasites were consistent with a pathway of protease trafficking from flagellar pocket to the lysosome/endosome compartment. The inhibitors were sufficiently absorbed and stable in vivo to ameliorate the pathology associated with a mouse model of Leishmania infection.

Pathophysiological significance and therapeutic applications of snake venom protease inhibitors.

PubMed

Thakur, Rupamoni; Mukherjee, Ashis K

2017-06-01

Protease inhibitors are important constituents of snake venom and play important roles in the pathophysiology of snakebite. Recently, research on snake venom protease inhibitors has provided valuable information to decipher the molecular details of various biological processes and offer insight for the development of some therapeutically important molecules from snake venom. The process of blood coagulation and fibrinolysis, in addition to affecting platelet function, are well known as the major targets of several snake venom protease inhibitors. This review summarizes the structure-functional aspects of snake venom protease inhibitors that have been described to date. Because diverse biological functions have been demonstrated by protease inhibitors, a comparative overview of their pharmacological and pathophysiological properties is also highlighted. In addition, since most snake venom protease inhibitors are non-toxic on their own, this review evaluates the different roles of individual protease inhibitors that could lead to the identification of drug candidates and diagnostic molecules. Copyright © 2017 Elsevier Ltd. All rights reserved.

Responsiveness of two Persian-versions of shoulder outcome measures following physiotherapy intervention in patients with shoulder disorders.

PubMed

Negahban, Hossein; Behtash, Zeinab; Sohani, Soheil Mansour; Salehi, Reza

2015-01-01

To identify the ability of the Persian-version of the Shoulder Pain and Disability Index (SPADI) and the Disabilities of the Arm, Shoulder, and Hand (DASH) to detect changes in shoulder function following physiotherapy intervention (i.e. responsiveness) and to determine the change score that indicates a meaningful change in functional ability of the patient (i.e. Minimally Clinically Important Difference (MCID)). A convenient sample of 200 Persian-speaking patients with shoulder disorders completed the SPADI and the DASH at baseline and then again 4 weeks after physiotherapy intervention. Furthermore, patients were asked to rate their global rating of shoulder function at follow-up. The responsiveness was evaluated using two methods: the receiver operating characteristics (ROC) method and the correlation analysis. Two useful statistics extracted from the ROC method are the area under curve (AUC) and the optimal cutoff point called as MCID. Both the SPADI and the DASH showed the AUC of greater than 0.70 (AUC ranges = 0.77-0.82). The best cutoff points (or change scores) for the SPADI-total, SPADI-pain, SPADI-disability and the DASH were 14.88, 26.36, 23.86, and 25.41, respectively. Additionally, moderate to good correlations (Gamma = -0.51 to -0.58) were found between the changes in SPADI/DASH and changes in global rating scale. The Persian SPADI and DASH have adequate responsiveness to clinical changes in patients with shoulder disorders. Moreover, the MCIDs obtained in this study will help the clinicians and researchers to determine if a Persian-speaking patient with shoulder disorder has experienced a true change following a physiotherapy intervention. Implications for Rehabilitation Responsiveness was evaluated using two methods; the receiver operating characteristics (ROC) method and the correlation analysis. The Persian SPADI and DASH can be used as two responsive instruments in both clinical practice and research settings. The MCIDs of 14.88 and 25.41 points obtained for the SPADI-total and DASH indicated that the change scores of at least 14.88 points on the SPADI-total and 25.41 points on the DASH is necessary to certain that a true change has occurred following a physiotherapy intervention.

Mobile Apps for the Dietary Approaches to Stop Hypertension (DASH): App Quality Evaluation.

PubMed

DiFilippo, Kristen Nicole; Huang, Wen-Hao David; Chapman-Novakofski, Karen M

2018-03-08

To identify the availability and quality of apps supporting Dietary Approaches to Stop Hypertension (DASH) education. The researchers identified DASH apps over 1 month in the Apple App Store. Five registered dietitians used the App Quality Evaluation (AQEL) to evaluate app quality on 7 domains. Interrater reliability was tested using intraclass correlations. One paid and 3 free DASH apps were evaluated. Interrater reliability (n = 5) was good for 3 apps and fair for 1 app. Only the paid app scored high (>8 of 10) on most AQEL quality domains. Based on lower quality found among the included free apps, further development of free apps is warranted. Whereas the paid app may be useful in supporting DASH education, future research should determine whether improvements in clinical outcomes are found and whether this app should be improved to address AQEL domains better. Copyright © 2018 Society for Nutrition Education and Behavior. Published by Elsevier Inc. All rights reserved.

The push-off test: development of a simple, reliable test of upper extremity weight-bearing capability.

PubMed

Vincent, Joshua I; MacDermid, Joy C; Michlovitz, Susan L; Rafuse, Richard; Wells-Rowsell, Christina; Wong, Owen; Bisbee, Leslie

2014-01-01

Longitudinal clinical measurement study. The push-off test (POT) is a novel and simple measure of upper extremity weight-bearing that can be measured with a grip dynamometer. There are no published studies on the validity and reliability of the POT. The relationship between upper extremity self-report activity/participation and impairment measures remain an unexplored realm. The primary purpose of this study is to estimate the intra and inter-rater reliability and construct validity of the POT. The secondary purpose is to estimate the relationship between upper extremity self-report activity/participation questionnaires and impairment measures. A convenience sample of 22 patients with wrist or elbow injuries were tested for POT, wrist/elbow range of motion (ROM), isometric wrist extension strength (WES) and grip strength; and completed two self-report activity/participation questionnaires: Disability of the Arm, Shoulder and the Hand (DASH) and Work Limitations Questionnaire (WLQ-26). POT's inter and intra-rater reliability and construct validity was tested. Pearson's correlations were run between the impairment measures and self-report questionnaires to look into the relationship amongst them. The POT demonstrated high inter-rater reliability (ICC affected = 0.97; 95% C.I. 0.93-0.99; ICC unaffected = 0.85; 95% C.I. 0.68-0.94) and intra-rater reliability (ICC affected = 0.96; 95% C.I. 0.92-0.97; ICC unaffected = 0.92; 95% C.I. 0.85-0.97). The POT was correlated moderately with the DASH (r = -0.47; p = 0.03). While examining the relationship between upper extremity self-reported activity/participation questionnaires and impairment measures the strongest correlation was between the DASH and the POT (r = -0.47; p = 0.03) and none of the correlations with the other physical impairment measures reached significance. At-work disability demonstrated insignificant correlations with physical impairments. The POT test provides a reliable and easily administered quantitative measure of ability to bear the load through an injured arm. Preliminary evidence supports a moderate relationship between loading bearing measured by the POT and upper extremity function measured by the DASH. 1b. Copyright © 2014 Hanley & Belfus. Published by Elsevier Inc. All rights reserved.

Cross-cultural adaptation of the Disability of Arm, Shoulder, and Hand questionnaire: Spanish for Puerto Rico Version

PubMed Central

Mulero-Portela, Ana L.; Colón-Santaella, Carmen L.; Cruz-Gomez, Cynthia

2010-01-01

The purpose of this study was to perform a cross-cultural adaptation of the Disability of Arm, Shoulder, and Hand (DASH) questionnaire to Spanish for Puerto Rico. Five steps were followed for the cross-cultural adaptation: forward translations into Spanish for Puerto Rico, synthesis of the translations, back translations into English, revision by an expert committee, and field test of the prefinal version. Psychometric characteristics of reliability and construct validity were evaluated for the final version. Internal consistency of the final version was high (Cronbach's α = 0.97) and item-to-total correlations were moderate (range from 0.44 to 0.85). Construct validity was evaluated by correlating the DASH with the scales of the Functional Assessment of Cancer Therapy - Breast. Fair to moderate correlations found in this study between the DASH and most scales of the Functional Assessment of Cancer Therapy - Breast support the construct validity of the Puerto Rico-Spanish DASH. The final version of the questionnaire was revised and approved by the Institute for Work and Health of Canada. Revisions to the original DASH English version are recommended. This version of the DASH is valid and reliable, and it can be used to evaluate outcomes in both clinical and research settings. PMID:19901616

Multiple Classes of Immune-Related Proteases Associated with the Cell Death Response in Pepper Plants

PubMed Central

Bae, Chungyun; Kim, Su-min; Lee, Dong Ju; Choi, Doil

2013-01-01

Proteases regulate a large number of biological processes in plants, such as metabolism, physiology, growth, and defense. In this study, we carried out virus-induced gene silencing assays with pepper cDNA clones to elucidate the biological roles of protease superfamilies. A total of 153 representative protease genes from pepper cDNA were selected and cloned into a Tobacco rattle virus-ligation independent cloning vector in a loss-of-function study. Silencing of 61 proteases resulted in altered phenotypes, such as the inhibition of shoot growth, abnormal leaf shape, leaf color change, and lethality. Furthermore, the silencing experiments revealed that multiple proteases play a role in cell death and immune response against avirulent and virulent pathogens. Among these 153 proteases, 34 modulated the hypersensitive cell death response caused by infection with an avirulent pathogen, and 16 proteases affected disease symptom development caused by a virulent pathogen. Specifically, we provide experimental evidence for the roles of multiple protease genes in plant development and immune defense following pathogen infection. With these results, we created a broad sketch of each protease function. This information will provide basic information for further understanding the roles of the protease superfamily in plant growth, development, and defense. PMID:23696830

Functional incapacity related to rotator cuff syndrome in workers. Is it influenced by social characteristics and medical management?

PubMed

Champagne, Romain; Bodin, Julie; Fouquet, Natacha; Roquelaure, Yves; Petit, Audrey

2017-12-04

Survey. Rotator cuff syndrome (RCS) is one of the most common musculoskeletal disorders reported in workers. The functional incapacity related to RCS may vary according to the sociodemographic context and to the medical management. The purpose of this is to analyze the RCS-related functional incapacity assessed by the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaires in workers according to their sociodemographic characteristics and the use of care. A cross-sectional study was carried out on a French sample of workers diagnosed with RCS. The DASH and DASH-work scores were studied according to the sociodemographic factors, musculoskeletal symptoms, and RCS medical management during the preceding 12 months. Two hundred seven workers who suffered from RCS filled out the questionnaire of which 80% were still working. The DASH score was significantly higher in women (24.0 vs 17.4; P < .01; effect size (d) = 0.39), in patients over the age of 50 years (23.6 vs 11.3; P < .005) and in case of another upper limb musculoskeletal disorder (P < .0001; d ≥ 0.4). The DASH and DASH-work scores were significantly higher in case of use of care for RCS (P < .005; d > 0.6). The demographic factors and the RCS medical management influenced the overall incapacity assessed by the DASH questionnaire. Work incapacity was more especially related to the use of care for RCS. The sociodemographic and medical parameters added to other established predictors could help guide clinicians in managing their patients. Copyright © 2017 Hanley & Belfus. Published by Elsevier Inc. All rights reserved.

DASH diet, insulin resistance, and serum hs-CRP in polycystic ovary syndrome: a randomized controlled clinical trial.

PubMed

Asemi, Z; Esmaillzadeh, A

2015-03-01

This study was designed to assess the effects of Dietary Approaches to Stop Hypertension (DASH) eating plan on insulin resistance and serum hs-CRP in overweight and obese women with PCOS. This randomized controlled clinical trial was done on 48 women diagnosed with PCOS. Subjects were randomly assigned to consume either the control (n=24) or the DASH eating pattern (n=24) for 8 weeks. The DASH diet consisted of 52% carbohydrates, 18% proteins, and 30% total fats. It was designed to be rich in fruits, vegetables, whole grains, and low-fat dairy products and low in saturated fats, cholesterol, refined grains, and sweets. Sodium content of the DASH diet was designed to be less than 2 400 mg/day. The control diet was also designed to contain 52% carbohydrates, 18% protein, and 30% total fat. Fasting blood samples were taken at baseline and after 8 weeks intervention to measure -insulin resistance and serum hs-CRP levels. -Adherence to the DASH eating pattern, compared to the -control diet, resulted in a significant reduction of serum insulin levels (-1.88 vs. 2.89 μIU/ml, p=0.03), HOMA-IR score (-0.45 vs. 0.80; p=0.01), and serum hs-CRP levels (-763.29 vs. 665.95 ng/ml, p=0.009). Additionally, a significant reduction in waist (-5.2 vs. -2.1 cm; p=0.003) and hip circumference (-5.9 vs. -1 cm; p<0.0001) was also seen in the DASH group compared with the control group. In conclusion, consumption of the DASH eating pattern for 8 weeks in overweight and obese women with PCOS resulted in the improvement of insulin resistance, serum hs-CRP levels, and abdominal fat accumulation. www.irct.ir: IRCT201304235623N6. © Georg Thieme Verlag KG Stuttgart · New York.

The association of the dietary approach to stop hypertension (DASH) diet with blood pressure, glucose and lipid profiles in Malaysian and Philippines populations.

PubMed

Tiong, X T; Nursara Shahirah, A; Pun, V C; Wong, K Y; Fong, A Y Y; Sy, R G; Castillo-Carandang, N T; Nang, E E K; Woodward, M; van Dam, R M; Tai, E S; Venkataraman, K

2018-05-07

Despite a growing body of evidence from Western populations on the health benefits of Dietary Approaches to Stop Hypertension (DASH) diets, their applicability in South East Asian settings is not clear. We examined cross-sectional associations between DASH diet and cardio-metabolic risk factors among 1837 Malaysian and 2898 Philippines participants in a multi-national cohort. Blood pressures, fasting lipid profile and fasting glucose were measured, and DASH score was computed based on a 22-item food frequency questionnaire. Older individuals, women, those not consuming alcohol and those undertaking regular physical activity were more likely to have higher DASH scores. In the Malaysian cohort, while total DASH score was not significantly associated with cardio-metabolic risk factors after adjusting for confounders, significant associations were observed for intake of green vegetable [0.011, standard error (SE): 0.004], and red and processed meat (-0.009, SE: 0.004) with total cholesterol. In the Philippines cohort, a 5-unit increase in total DASH score was significantly and inversely associated with systolic blood pressure (-1.41, SE: 0.40), diastolic blood pressure (-1.09, SE: 0.28), total cholesterol (-0.015, SE: 0.005), low-density lipoprotein cholesterol (-0.025, SE: 0.008), and triglyceride (-0.034, SE: 0.012) after adjusting for socio-demographic and lifestyle groups. Intake of milk and dairy products, red and processed meat, and sugared drinks were found to significantly associated with most risk factors. Differential associations of DASH diet and dietary components with cardio-metabolic risk factors by country suggest the need for country-specific tailoring of dietary interventions to improve cardio-metabolic risk profiles. Copyright © 2018 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.

The use of a tablet computer to complete the DASH questionnaire.

PubMed

Dy, Christopher J; Schmicker, Thomas; Tran, Quynh; Chadwick, Brian; Daluiski, Aaron

2012-12-01

To determine whether electronic self-administration of the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire using a tablet computer increased completion rate compared with paper self-administration. We gave the DASH in self-administered paper form to 222 new patients in a single hand surgeon's practice. After a washout period of 5 weeks, we gave the DASH in self-administered tablet computer form to 264 new patients. A maximum of 3 questions could be omitted before the questionnaire was considered unscorable. We reviewed the submitted surveys to determine the number of scorable questionnaires and the number of omitted questions in each survey. We completed univariate analysis and regression modeling to determine the influence of survey administration type on respondent error while controlling for patient age and sex. Of the 486 total surveys, 60 (12%) were not scorable. A significantly higher proportion of the paper surveys (24%) were unscorable compared with electronic surveys (2%), with significantly more questions omitted in each paper survey (2.6 ± 4.4 questions) than in each electronic survey (0.1 ± 0.8 questions). Logistic regression analysis revealed survey administration mode to be significantly associated with DASH scorability while controlling for age and sex, with electronic survey administration being 14 times more likely than paper administration to yield a scorable DASH. In our retrospective series, electronic self-administration of the DASH decreased the number of omitted questions and yielded a higher number of scorable questionnaires. Prospective, randomized evaluation is needed to better delineate the effect of survey administration on respondent error. Administration of the DASH with a tablet computer may be beneficial for both clinical and research endeavors to increase completion rate and to gain other benefits from electronic data capture. Copyright © 2012 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.

Time Course of Change in Blood Pressure From Sodium Reduction and the DASH Diet.

PubMed

Juraschek, Stephen P; Woodward, Mark; Sacks, Frank M; Carey, Vincent J; Miller, Edgar R; Appel, Lawrence J

2017-11-01

Both sodium reduction and the Dietary Approaches to Stop Hypertension (DASH) diet lower blood pressure (BP); however, the patterns of their effects on BP over time are unknown. In the DASH-Sodium trial, adults with pre-/stage 1 hypertension, not using antihypertensive medications, were randomly assigned to either a typical American diet (control) or DASH. Within their assigned diet, participants randomly ate each of 3 sodium levels (50, 100, and 150 mmol/d, at 2100 kcal) over 4-week periods. BP was measured weekly for 12 weeks; 412 participants enrolled (57% women; 57% black; mean age, 48 years; mean systolic BP [SBP]/diastolic BP [DBP], 135/86 mm Hg). For those assigned control, there was no change in SBP/DBP between weeks 1 and 4 on the high-sodium diet (weekly change, -0.04/0.06 mm Hg/week) versus a progressive decline in BP on the low-sodium diet (-0.94/-0.70 mm Hg/week; P interactions between time and sodium <0.001 for SBP and DBP). For those assigned DASH, SBP/DBP changed -0.60/-0.16 mm Hg/week on the high- versus -0.42/-0.54 mm Hg/week on the low-sodium diet ( P interactions between time and sodium=0.56 for SBP and 0.10 for DBP). When comparing DASH to control, DASH changed SBP/DBP by -4.36/-1.07 mm Hg after 1 week, which accounted for most of the effect observed, with no significant difference in weekly rates of change for either SBP ( P interaction=0.97) or DBP ( P interaction=0.70). In the context of a typical American diet, a low-sodium diet reduced BP without plateau, suggesting that the full effects of sodium reduction are not completely achieved by 4 weeks. In contrast, compared with control, DASH lowers BP within a week without further effect thereafter. URL: http://www.clinicaltrials.gov. Unique identifier: NCT00000608. © 2017 American Heart Association, Inc.

An alternative approach to depigmentation by soybean extracts via inhibition of the PAR-2 pathway.

PubMed

Paine, C; Sharlow, E; Liebel, F; Eisinger, M; Shapiro, S; Seiberg, M

2001-04-01

The protease-activated receptor 2, expressed on keratinocytes but not on melanocytes, has been ascribed functional importance in the regulation of pigmentation by phagocytosis of melanosomes. Inhibition of protease-activated receptor 2 activation by synthetic serine protease inhibitors requires keratinocyte-melanocyte contact and results in depigmentation of the dark skinned Yucatan swine, suggesting a new class of depigmenting mechanism and agents. We therefore examined natural agents that could exert their effect via the protease-activated receptor 2 pathway. Here we show that soymilk and the soybean-derived serine protease inhibitors soybean trypsin inhibitor and Bowman-Birk inhibitor inhibit protease-activated receptor 2 cleavage, affect cytoskeletal and cell surface organization, and reduce keratinocyte phagocytosis. The depigmenting activity of these agents and their capability to prevent ultraviolet-induced pigmentation are demonstrated both in vitro and in vivo. These results imply that inhibition of the protease-activated receptor 2 pathway by soymilk may be used as a natural alternative to skin lightening.

Structural and Functional Characterization of Cleavage and Inactivation of Human Serine Protease Inhibitors by the Bacterial SPATE Protease EspPα from Enterohemorrhagic E. coli

PubMed Central

Weiss, André; Joerss, Hanna; Brockmeyer, Jens

2014-01-01

EspPα and EspI are serine protease autotransporters found in enterohemorrhagic Escherichia coli. They both belong to the SPATE autotransporter family and are believed to contribute to pathogenicity via proteolytic cleavage and inactivation of different key host proteins during infection. Here, we describe the specific cleavage and functional inactivation of serine protease inhibitors (serpins) by EspPα and compare this activity with the related SPATE EspI. Serpins are structurally related proteins that regulate vital protease cascades, such as blood coagulation and inflammatory host response. For the rapid determination of serpin cleavage sites, we applied direct MALDI-TOF-MS or ESI-FTMS analysis of coincubations of serpins and SPATE proteases and confirmed observed cleavage positions using in-gel-digest of SDS-PAGE-separated degradation products. Activities of both serpin and SPATE protease were assessed in a newly developed photometrical assay using chromogenic peptide substrates. EspPα cleaved the serpins α1-protease inhibitor (α1-PI), α1-antichymotrypsin, angiotensinogen, and α2-antiplasmin. Serpin cleavage led to loss of inhibitory function as demonstrated for α1-PI while EspPα activity was not affected. Notably, EspPα showed pronounced specificity and cleaved procoagulatory serpins such as α2-antiplasmin while the anticoagulatory antithrombin III was not affected. Together with recently published research, this underlines the interference of EspPα with hemostasis or inflammatory responses during infection, while the observed interaction of EspI with serpins is likely to be not physiologically relevant. EspPα-mediated serpin cleavage occurred always in flexible loops, indicating that this structural motif might be required for substrate recognition. PMID:25347319

Mediterranean diet, Dietary Approaches to Stop Hypertension (DASH) style diet, and metabolic health in U.S. adults.

PubMed

Park, Yong-Moon Mark; Steck, Susan E; Fung, Teresa T; Zhang, Jiajia; Hazlett, Linda J; Han, Kyungdo; Lee, Seung-Hwan; Kwon, Hyuk-Sang; Merchant, Anwar T

2017-10-01

There is sparse evidence on the relationship between the Mediterranean diet, Dietary Approaches to Stop Hypertension (DASH) style diet, and metabolic health, especially comparing cardiometabolic phenotypes among in normal weight and obese populations. We aimed to investigate the association of the Mediterranean diet scores (MDS) and DASH index with metabolically healthy obese (MHO) and metabolically obese normal weight (MONW) phenotypes in a representative U.S. MDS and DASH index were calculated using dietary data from 2767 adults aged 20-90 years without any prior diagnosis of cancer or cardiovascular disease from the National Health and Nutrition Examination Survey III, 1988-1994. MHO and MONW individuals were identified using fasting glucose, insulin resistance, blood pressure, triglycerides, C-reactive protein, and high-density lipoprotein-cholesterol. Higher MDS was associated with higher odds of MHO phenotype (odds ratio (OR) T3 vs T1 , 2.57 [95% confidence interval (CI), 1.04-6.35]; P trend = 0.04), and higher DASH index was associated with lower odds of MONW phenotype (OR T3 vs T1, 0.59 [95% CI, 0.38-0.93]; P trend = 0.03) only in the younger age group (<45 years for men or premenopausal women). No significant associations of MDS and DASH index with MHO and MONW phenotypes were observed in the older age group (≥45 years for men or postmenopausal women). Adherence to Mediterranean diet or DASH style diet was favorably associated with MHO and MONW phenotypes only in the younger age group, suggesting that potential dietary intervention to prevent cardiometabolic disease differ by age group. Published by Elsevier Ltd.

Pacific kids DASH for health (PacDASH) randomized, controlled trial with DASH eating plan plus physical activity improves fruit and vegetable intake and diastolic blood pressure in children.

PubMed

Novotny, Rachel; Nigg, Claudio R; Li, Fenfang; Wilkens, Lynne R

2015-04-01

Pacific Kids DASH for Health (PacDASH) aimed to improve child diet and physical activity (PA) level and prevent excess weight gain and elevation in blood pressure (BP) at 9 months. PacDASH was a two-arm, randomized, controlled trial (ClinicalTrials.gov: NCT00905411). Eighty-five 5- to 8-year-olds in the 50th-99th percentile for BMI were randomly assigned to treatment (n=41) or control (n=44) groups; 62 completed the 9-month trial. Sixty-two percent were female. Mean age was 7.1±0.95 years. Race/ethnicity was Asian (44%), Native Hawaiian or Other Pacific Islander (28%), white (21%), or other race/ethnicity (7%). Intervention was provided at baseline and 3, 6 and 9 months, with monthly supportive mailings between intervention visits, and a follow-up visit at 15 months to observe maintenance. Diet and PA were assessed by 2-day log. Body size, composition, and BP were measured. The intervention effect on diet and PA, body size and composition, and BP by the end of the intervention was tested using an F test from a mixed regression model, after adjustment for sex, age, and ethnic group. Fruit and vegetable (FV) intake decreased less in the treatment than control group (p=0.04). Diastolic BP (DBP) was 12 percentile units lower in the treatment than control group after 9 months of intervention (p=0.01). There were no group differences in systolic BP (SBP) or body size/composition. The PacDASH trial enhanced FV intake and DBP, but not SBP or body size/composition.

Development of a QSAR model for predicting aqueous reaction rate constants of organic chemicals with hydroxyl radicals.

PubMed

Luo, Xiang; Yang, Xianhai; Qiao, Xianliang; Wang, Ya; Chen, Jingwen; Wei, Xiaoxuan; Peijnenburg, Willie J G M

2017-03-22

Reaction with hydroxyl radicals (˙OH) is an important removal pathway for organic pollutants in the aquatic environment. The aqueous reaction rate constant (k OH ) is therefore an important parameter for fate assessment of aquatic pollutants. Since experimental determination fails to meet the requirement of being able to efficiently handle numerous organic chemicals at limited cost and within a relatively short period of time, in silico methods such as quantitative structure-activity relationship (QSAR) models are needed to predict k OH . In this study, a QSAR model with a larger and wider applicability domain as compared with existing models was developed. Following the guidelines for the development and validation of QSAR models proposed by the Organization for Economic Co-operation and Development (OECD), the model shows satisfactory performance. The applicability domain of the model has been extended and contained chemicals that have rarely been covered in most previous studies. The chemicals covered in the current model contain functional groups including [double bond splayed left]C[double bond, length as m-dash]C[double bond splayed right], -C[triple bond, length as m-dash]C-, -C 6 H 5 , -OH, -CHO, -O-, [double bond splayed left]C[double bond, length as m-dash]O, -C[double bond, length as m-dash]O(O)-, -COOH, -C[triple bond, length as m-dash]N, [double bond splayed left]N-, -NH 2 , -NH-C(O)-, -NO 2 , -N[double bond, length as m-dash]C-N[double bond splayed right], [double bond splayed left]N-N[double bond splayed right], -N[double bond, length as m-dash]N-, -S-, -S-S-, -SH, -SO 3 , -SO 4 , -PO 4 , and -X (F, Cl, Br, and I).

Accessibility and Affordability of Supermarkets: Associations With the DASH Diet.

PubMed

Mackenbach, Joreintje D; Burgoine, Thomas; Lakerveld, Jeroen; Forouhi, Nita G; Griffin, Simon J; Wareham, Nicholas J; Monsivais, Pablo

2017-07-01

It is unknown whether there is an interplay of affordability (economic accessibility) and proximity (geographic accessibility) of supermarkets in relation to having a Dietary Approaches to Stop Hypertension (DASH)-accordant diet. Data (collected: 2005-2015, analyzed: 2016) were from the cross-sectional, population-based Fenland Study cohort: 9,274 adults aged 29-64 years, living in Cambridgeshire, United Kingdom. Dietary quality was evaluated using an index of DASH dietary accordance, based on recorded consumption of foods and beverages in a validated 130-item, semi-quantitative food frequency questionnaire. DASH accordance was defined as a DASH score in the top quintile. Dietary costs (£/day) were estimated by attributing a food price variable to the foods consumed according to the questionnaire. Individuals were classified as having low-, medium-, or high-cost diets. Supermarket affordability was determined based on the cost of a 101-item market basket. Distances between home address to the nearest supermarket (geographic accessibility) and nearest economically-appropriate supermarket (economic accessibility) were divided into tertiles. Higher-cost diets were more likely to be DASH-accordant. After adjustment for key demographics and exposure to other food outlets, individuals with lowest economic accessibility to supermarkets had lower odds of being DASH-accordant (OR=0.59, 95% CI=0.52, 0.68) than individuals with greatest economic accessibility. This association was stronger than with geographic accessibility alone (OR=0.85, 95% CI=0.74, 0.98). Results suggest that geographic and economic access to food should be taken into account when considering approaches to promote adherence to healthy diets for the prevention of cardiovascular diseases and other chronic disease. Copyright © 2017 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights reserved.

High performance mode locking characteristics of single section quantum dash lasers.

PubMed

Rosales, Ricardo; Murdoch, S G; Watts, R T; Merghem, K; Martinez, Anthony; Lelarge, Francois; Accard, Alain; Barry, L P; Ramdane, Abderrahim

2012-04-09

Mode locking features of single section quantum dash based lasers are investigated. Particular interest is given to the static spectral phase profile determining the shape of the mode locked pulses. The phase profile dependence on cavity length and injection current is experimentally evaluated, demonstrating the possibility of efficiently using the wide spectral bandwidth exhibited by these quantum dash structures for the generation of high peak power sub-picosecond pulses with low radio frequency linewidths.

Validation of Ocean Color Sensors Using a Profiling Hyperspectral Radiometer

DTIC Science & Technology

2014-01-01

shadows. The HyperOCRs are all thermally characterized for temperature corrections and spectrally characterized to account for stray light corrections...August 24,2010 is shown in Figure 4A along with the mean percent difference between the NOAA Hyperpro ( Black /Dash) and the other two identical Hyperpro...difference (n=24) between the NOAA Hyperpro ( Black /Dash, Fig. 4A) and the other two Hyperpro systems. The dotted line for the red (bottom) and dash line for

Dentilisin activity affects the organization of the outer sheath of Treponema denticola.

PubMed

Ishihara, K; Kuramitsu, H K; Miura, T; Okuda, K

1998-08-01

Prolyl-phenylalanine-specific serine protease (dentilisin) is a major extracellular protease produced by Treponema denticola. The gene, prtP, coding for the protease was recently cloned and sequenced (K. Ishihara, T. Miura, H. K. Kuramitsu, and K. Okuda, Infect. Immun. 64:5178-5186, 1996). In order to determine the role of this protease in the physiology and virulence of T. denticola, a dentilisin-deficient mutant, K1, was constructed following electroporation with a prtP-inactivated DNA fragment. No chymotrypsin-like protease activity was detected in the dentilisin-deficient mutant. In addition, the high-molecular-mass oligomeric protein characteristic of the outer sheath of the organism decreased in the mutant. Furthermore, the hydrophobicity of the mutant was decreased, and coaggregation of the mutant with Fusobacterium nucleatum was enhanced compared to that of the wild-type organism. The results obtained with a mouse abscess model system indicated that the virulence of the mutant was attenuated relative to that of the wild-type organism. These results suggest that dentilisin activity plays a major role in the structural organization of the outer sheath of T. denticola. The loss of dentilsin activity and the structural change in the outer sheath affect the pathogenicity of T. denticola.

High performance dash on warning air mobile, missile system. [intercontinental ballistic missiles - systems analysis

NASA Technical Reports Server (NTRS)

Levin, A. D.; Castellano, C. R.; Hague, D. S.

1975-01-01

An aircraft-missile system which performs a high acceleration takeoff followed by a supersonic dash to a 'safe' distance from the launch site is presented. Topics considered are: (1) technological feasibility to the dash on warning concept; (2) aircraft and boost trajectory requirements; and (3) partial cost estimates for a fleet of aircraft which provide 200 missiles on airborne alert. Various aircraft boost propulsion systems were studied such as an unstaged cryogenic rocket, an unstaged storable liquid, and a solid rocket staged system. Various wing planforms were also studied. Vehicle gross weights are given. The results indicate that the dash on warning concept will meet expected performance criteria, and can be implemented using existing technology, such as all-aluminum aircraft and existing high-bypass-ratio turbofan engines.

Secretory leukocyte protease inhibitor protein regulates the penetrance of frontotemporal lobar degeneration in progranulin mutation carriers.

PubMed

Ghidoni, Roberta; Flocco, Rosa; Paterlini, Anna; Glionna, Michela; Caruana, Loredana; Tonoli, Elisa; Binetti, Giuliano; Benussi, Luisa

2014-01-01

The discovery that mutations in the gene encoding for progranulin (GRN) cause frontotemporal lobar degeneration (FTLD) and other neurodegenerative diseases leading to dementia has brought renewed interest in progranulin and its functions in the central nervous system. Full length progranulin is preserved from cleavage by secretory leukocyte protease inhibitor (SLPI), one of the smallest serine protease inhibitor circulating in plasma. Herein, we investigated the relationship between circulating SLPI and progranulin in affected and unaffected subjects belonging to 26 Italian pedigrees carrying GRN null mutations. In GRN null mutation carriers, we demonstrated: i) an increase of circulating SLPI levels in affected subjects; ii) an age-related upregulation of the serine-protease inhibitor in response to lifetime progranulin shortage; and iii) a delay in the age of onset in subjects with the highest SLPI protein levels. The study of SLPI and its relation to progranulin suggests the existence of unexpected molecular players in progranulin-associated neurodegeneration.

DASH Eating Plan

MedlinePlus

... Approaches to Stop Hypertension. It is an eating plan that is based on research studies sponsored by ... risk of getting heart disease. The DASH eating plan Emphasizes vegetables, fruits, and whole-grains Includes fat- ...

Facile synthesis of -C[double bond, length as m-dash]N- linked covalent organic frameworks under ambient conditions.

PubMed

Ding, San-Yuan; Cui, Xiao-Hui; Feng, Jie; Lu, Gongxuan; Wang, Wei

2017-10-31

We reported herein a facile approach for the synthesis of -C[double bond, length as m-dash]N- linked covalent organic frameworks under ambient conditions. Three known (COF-42, COF-43, and COF-LZU1) and one new (Pr-COF-42) COF materials were successfully synthesized using this method. Furthermore, this simple synthetic approach makes the large-scale synthesis of -C[double bond, length as m-dash]N- linked COFs feasible.

Materials and Modulators for 3D Displays

DTIC Science & Technology

2002-08-01

1243 nm. 0, 180 and 360 deg. in this plot correspond to parallel polarization. The dashed curve is a cos2(θ) fit to the data with a constant value...dwell time (solid bold curve ), 10 µs dwell time (dashed bold curve ) and static case (thin dashed curve ). 26 Figure. 20. Schematics of free-space...photon. The two peaks in the two photon spectrum can be fit by two Lorentzian curves . These spectra indicate that in the rhodamine B molecule the

Identification of the formation of metal-vinylidene interfacial bonds of alkyne-capped platinum nanoparticles by isotopic labeling.

PubMed

Hu, Peiguang; Chen, Limei; Deming, Christopher P; Bonny, Lewis W; Lee, Hsiau-Wei; Chen, Shaowei

2016-10-07

Stable platinum nanoparticles were prepared by the self-assembly of 1-dodecyne and dodec-1-deuteroyne onto bare platinum colloid surfaces. The nanoparticles exhibited consistent core size and optical properties. FTIR and NMR measurements confirmed the formation of Pt-vinylidene (Pt[double bond, length as m-dash]C[double bond, length as m-dash]CH-) interfacial linkages rather than Pt-acetylide (Pt-C[triple bond, length as m-dash]C-) and platinum-hydride (Pt-H) bonds.

The Dietary Approaches to Stop Hypertension Diet, Cognitive Function, and Cognitive Decline in American Older Women.

PubMed

Berendsen, Agnes A M; Kang, Jae H; van de Rest, Ondine; Feskens, Edith J M; de Groot, Lisette C P G M; Grodstein, Francine

2017-05-01

To examine the association between long-term adherence to the Dietary Approaches to Stop Hypertension (DASH) diet with cognitive function and decline in older American women. Prospective cohort study. The Nurses' Health Study, a cohort of registered nurses residing in 11 US states. A total of 16,144 women from the Nurses' Health Study, aged ≥70 years, who underwent cognitive testing a total of 4 times by telephone from 1995 to 2001 (baseline), with multiple dietary assessments between 1984 and the first cognitive examination. DASH adherence for each individual was based on scoring of intakes of 9 nutrient or food components. Long-term DASH adherence was calculated as the average DASH adherence score from up to 5 repeated measures of diet. Primary outcomes were cognitive function calculated as the average scores of the 4 repeated measures, as well as cognitive change of the Telephone Interview for Cognitive Status score and composite scores of global cognition and verbal memory. Greater adherence to long-term DASH score was associated with better average cognitive function, irrespective of apolipoprotein E ε4 allele status [multivariable-adjusted differences in mean z-scores between extreme DASH quintiles = 0.04 (95% confidence interval, CI 0.01-0.07), P trend = .009 for global cognition; 0.04 (95% CI 0.01-0.07), P trend = .002 for verbal memory and 0.16 (95% CI 0.03-0.29), and P trend = .03 for Telephone Interview for Cognitive Status, P interaction >0.24]. These differences were equivalent to being 1 year younger in age. Adherence to the DASH score was not associated with change in cognitive function over 6 years. Our findings in the largest cohort on dietary patterns and cognitive function to date indicate that long-term adherence to the DASH diet is important to maintain cognitive function at older ages. Copyright © 2016 AMDA – The Society for Post-Acute and Long-Term Care Medicine. All rights reserved.

Platelet-rich plasma (PRP) in chronic epicondylitis: study protocol for a randomized controlled trial.

PubMed

Martin, Jose I; Merino, Josu; Atilano, Leire; Areizaga, Luis M; Gomez-Fernandez, Maria C; Burgos-Alonso, Natalia; Andia, Isabel

2013-12-01

Tendinopathy is a difficult problem to manage and can result in significant patient morbidity. Currently, the clinical use of platelet-rich plasma (PRP) in painful tendons is widespread but its efficacy remains controversial. This study is a single-center, randomized double-blind controlled trial. Eighty patients will be allocated to have ultrasound (US)-guided needling combined with a leukocyte-depleted (that is, pure) PRP or lidocaine each alternate week for a total of two interventions. Outcome data will be collected before intervention, and at 6 weeks, 3, 6, and 12 months after intervention. Changes in pain and activity levels, as assessed by Disabilities of the Arm, Shoulder and Hand (DASH-E, Spanish version) score, at 6 months. We will compare the percentage of patients in each group that achieve a successful treatment defined as a reduction of at least 25% in the DASH-E score. Secondary outcome measures include changes in DASH-E at 3 and 12 months, changes in pain as assessed by the visual analogue scale (VAS) at the 6-week, 3-, 6-, and 12-month follow-up, changes in sonographic features and neovascularity, and percentage of patients in each group with adverse reactions at 3, 6, and 12 months. The results of this study will provide insights into the effect of pure PRP in tendon and may contribute to identifying the best protocol for PRP application in tendinopathies. ClinicalTrials.gov: NCT01945528.

Proteases from Entamoeba spp. and Pathogenic Free-Living Amoebae as Virulence Factors

PubMed Central

Serrano-Luna, Jesús; Piña-Vázquez, Carolina; Reyes-López, Magda; Ortiz-Estrada, Guillermo

2013-01-01

The standard reference for pathogenic and nonpathogenic amoebae is the human parasite Entamoeba histolytica; a direct correlation between virulence and protease expression has been demonstrated for this amoeba. Traditionally, proteases are considered virulence factors, including those that produce cytopathic effects in the host or that have been implicated in manipulating the immune response. Here, we expand the scope to other amoebae, including less-pathogenic Entamoeba species and highly pathogenic free-living amoebae. In this paper, proteases that affect mucin, extracellular matrix, immune system components, and diverse tissues and cells are included, based on studies in amoebic cultures and animal models. We also include proteases used by amoebae to degrade iron-containing proteins because iron scavenger capacity is currently considered a virulence factor for pathogens. In addition, proteases that have a role in adhesion and encystation, which are essential for establishing and transmitting infection, are discussed. The study of proteases and their specific inhibitors is relevant to the search for new therapeutic targets and to increase the power of drugs used to treat the diseases caused by these complex microorganisms. PMID:23476670

DASH Eating Plan

MedlinePlus

... Card Other Resources ChooseMyPlate.gov (U.S. Department of Agriculture, U.S. Department of Health and Human Services) DASH ( ... Dietary Guidelines for Americans, 2010 (U.S. Department of Agriculture, U.S. Department of Health and Human Services) High ...

Utilisation of an eta(3)-allyl hydride complex, formed by UV irradiation, as a controlled source of 16-electron (eta(5)-C(5)Me(5))Rh(CH(2)[double bond, length as m-dash]CHMe).

PubMed

Sexton, Catherine J; López-Serrano, Joaquín; Lledós, Agustí; Duckett, Simon B

2008-10-21

Low temperature UV irradiation of solutions of (eta(5)-C(5)Me(5))Rh(CH(2)[double bond, length as m-dash]CHMe)(2) yields (eta(5)-C(5)Me(5))Rh(eta(3)-CH(2)CHCH(2))(H), which provides controlled access to the 16-electron fragment (eta(5)-C(5)Me(5))Rh(CH(2)[double bond, length as m-dash]CHMe).

Dispersion and Attenuation Due to Scattering from Heterogeneities of the Frame Bulk Modulus of a Poroelastic Medium

DTIC Science & Technology

2010-02-19

attenuation is a function of the Hurst exponent which characterizes the fractal het- erogeneity. Muller and Gurevich15,16 used statistical smoothing of...modified Bessel function of the third kind, Γ denotes the gamma function, and ν is the Hurst coefficient which is assumed to be 0 < ν ≤ 1. The three...The Hurst coefficient, ν, is ν = 0.1 (long-dashed line), ν = 0.5 (short-dashed line), and ν = 0.9 (long-short dashed line). In (a) the sound speed

The relationship between protease/anti-protease profile, angiogenesis and re-epithelialisation in acute burn wounds.

PubMed

Caulfield, Robert H; Tyler, Michael P H; Austyn, Jon M; Dziewulski, Peter; McGrouther, Duncan A

2008-06-01

In the management of partial thickness burns, it is difficult to balance between conservative management and surgical intervention. Our hypothesis was that a triangular relationship exists between protease/anti-protease profile at the burn wound surface, angiogenesis and re-epithelialisation. By manipulation of the biochemical profile at the wound level, we determined to affect the nature and extent of angiogenesis and resulting re-epithelialisation. We performed a randomised longitudinal observational study on partial thickness burns in adult patients presenting to two regional burns units. Our results demonstrated that a high-protease wound environment is associated with lower levels of the angiogenic factor VEGF, a lower more uniform change in wound bloodflow and a uniform well healed wound with an architecturally normal epidermis. In addition, we found that a low protease wound environment is associated with higher levels of the angiogenic factor VEGF, a higher wound bloodflow throughout the wound healing period and a more chaotic, hypercellular, overkeratinised, and chaotic thickened epidermis.

Effects of Sodium Reduction and the DASH Diet in Relation to Baseline Blood Pressure.

PubMed

Juraschek, Stephen P; Miller, Edgar R; Weaver, Connie M; Appel, Lawrence J

2017-12-12

Both sodium reduction and the DASH (Dietary Approaches to Stop Hypertension) diet, a diet rich in fruits, vegetables, and low-fat dairy products, and reduced in saturated fat and cholesterol, lower blood pressure. The separate and combined effects of these dietary interventions by baseline blood pressure (BP) has not been reported. The authors compared the effects of low versus high sodium, DASH versus control, and both (low sodium-DASH vs. high sodium-control diets) on systolic blood pressure (SBP) by baseline BP. In the DASH-Sodium (Dietary Patterns, Sodium Intake and Blood Pressure) trial, adults with pre- or stage 1 hypertension and not using antihypertensive medications, were randomized to either DASH or a control diet. On either diet, participants were fed each of 3 sodium levels (50, 100, and 150 mmol/day at 2,100 kcal) in random order over 4 weeks separated by 5-day breaks. Strata of baseline SBP were <130, 130 to 139, 140 to 149, and ≥150 mm Hg. Of 412 participants, 57% were women, and 57% were black; mean age was 48 years, and mean SBP/diastolic BP was 135/86 mm Hg. In the context of the control diet, reducing sodium (from high to low) was associated with mean SBP differences of -3.20, -8.56, -8.99, and -7.04 mm Hg across the respective baseline SBP strata listed (p for trend = 0.004). In the context of high sodium, consuming the DASH compared with the control diet was associated with mean SBP differences of -4.5, -4.3, -4.7, and -10.6 mm Hg, respectively (p for trend = 0.66). The combined effects of the low sodium-DASH diet versus the high sodium-control diet on SBP were -5.3, -7.5, -9.7, and -20.8 mm Hg, respectively (p for trend <0.001). The combination of reduced sodium intake and the DASH diet lowered SBP throughout the range of pre- and stage 1 hypertension, with progressively greater reductions at higher levels of baseline SBP. SBP reductions in adults with the highest levels of SBP (≥150 mm Hg) were striking and reinforce the importance of both sodium reduction and the DASH diet in this high-risk group. Further research is needed to determine the effects of these interventions among adults with SBP ≥160 mm Hg. (Dietary Patterns, Sodium Intake and Blood Pressure [DASH-Sodium]; NCT00000608). Copyright © 2017 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.

Rasch Model Analysis Gives New Insights Into the Structural Validity of the QuickDASH in Patients With Musculoskeletal Shoulder Pain.

PubMed

Jerosch-Herold, Christina; Chester, Rachel; Shepstone, Lee

2017-09-01

Study Design Cross-sectional secondary analysis of a prospective cohort study. Background The shortened version of the Disabilities of the Arm, Shoulder and Hand questionnaire (QuickDASH) is a widely used outcome measure that has been extensively evaluated using classical test theory. Rasch model analysis can identify strengths and weaknesses of rating scales and goes beyond classical test theory approaches. It uses a mathematical model to test the fit between the observed data and expected responses and converts ordinal-level scores into interval-level measurement. Objective To test the structural validity of the QuickDASH using Rasch analysis. Methods A prospective cohort study of 1030 patients with shoulder pain provided baseline data. Rasch analysis was conducted to (1) assess how the QuickDASH fits the Rasch model, (2) identify sources of misfit, and (3) explore potential solutions to these. Results There was evidence of multidimensionality and significant misfit to the Rasch model (χ 2 = 331.09, P<.001). Two items had disordered threshold responses with strong floor effects. Response bias was detected in most items for age and sex. Rescoring resulted in ordered thresholds; however, the 11-item scale still did not meet the expectations of the Rasch model. Conclusion Rasch model analysis on the QuickDASH has identified a number of problems that cannot be easily detected using traditional analyses. While revisions to the QuickDASH resulted in better fit, a "shoulder-specific" version is not advocated at present. Caution needs to be exercised when interpreting results of the QuickDASH outcome measure, as it does not meet the criteria for interval-level measurement and shows significant response bias by age and sex. J Orthop Sports Phys Ther 2017;47(9):664-672. Epub 13 Jul 2017. doi:10.2519/jospt.2017.7288.

The Association of Mediterranean and DASH Diets with Mortality in Adults on Hemodialysis: The DIET-HD Multinational Cohort Study.

PubMed

Saglimbene, Valeria M; Wong, Germaine; Craig, Jonathan C; Ruospo, Marinella; Palmer, Suetonia C; Campbell, Katrina; Garcia-Larsen, Vanessa; Natale, Patrizia; Teixeira-Pinto, Armando; Carrero, Juan-Jesus; Stenvinkel, Peter; Gargano, Letizia; Murgo, Angelo M; Johnson, David W; Tonelli, Marcello; Gelfman, Rubén; Celia, Eduardo; Ecder, Tevfik; Bernat, Amparo G; Del Castillo, Domingo; Timofte, Delia; Török, Marietta; Bednarek-Skublewska, Anna; Duława, Jan; Stroumza, Paul; Hoischen, Susanne; Hansis, Martin; Fabricius, Elisabeth; Felaco, Paolo; Wollheim, Charlotta; Hegbrant, Jörgen; Strippoli, Giovanni F M

2018-06-01

Background Mediterranean and Dietary Approaches to Stop Hypertension (DASH) diets associate with lower cardiovascular and all-cause mortality in the general population, but the benefits for patients on hemodialysis are uncertain. Methods Mediterranean and DASH diet scores were derived from the GA 2 LEN Food Frequency Questionnaire within the DIET-HD Study, a multinational cohort study of 9757 adults on hemodialysis. We conducted adjusted Cox regression analyses clustered by country to evaluate the association between diet score tertiles and all-cause and cardiovascular mortality (the lowest tertile was the reference category). Results During the median 2.7-year follow-up, 2087 deaths (829 cardiovascular deaths) occurred. The adjusted hazard ratios (95% confidence intervals) for the middle and highest Mediterranean diet score tertiles were 1.20 (1.01 to 1.41) and 1.14 (0.90 to 1.43), respectively, for cardiovascular mortality and 1.10 (0.99 to 1.22) and 1.01 (0.88 to 1.17), respectively, for all-cause mortality. Corresponding estimates for the same DASH diet score tertiles were 1.01 (0.85 to 1.21) and 1.19 (0.99 to 1.43), respectively, for cardiovascular mortality and 1.03 (0.92 to 1.15) and 1.00 (0.89 to 1.12), respectively, for all-cause mortality. The association between DASH diet score and all-cause death was modified by age ( P =0.03); adjusted hazard ratios for the middle and highest DASH diet score tertiles were 1.02 (0.81 to 1.29) and 0.70 (0.53 to 0.94), respectively, for younger patients (≤60 years old) and 1.05 (0.93 to 1.19) and 1.08 (0.95 to 1.23), respectively, for older patients. Conclusions Mediterranean and DASH diets did not associate with cardiovascular or total mortality in hemodialysis. Copyright © 2018 by the American Society of Nephrology.

Effects of dietary sodium and the DASH diet on the occurrence of headaches: results from randomised multicentre DASH-Sodium clinical trial.

PubMed

Amer, Muhammad; Woodward, Mark; Appel, Lawrence J

2014-12-11

Headaches are a common medical problem, yet few studies, particularly trials, have evaluated therapies that might prevent or control headaches. We, thus, investigated the effects on the occurrence of headaches of three levels of dietary sodium intake and two diet patterns (the Dietary Approaches to Stop Hypertension (DASH) diet (rich in fruits, vegetables and low-fat dairy products with reduced saturated and total fat) and a control diet (typical of Western consumption patterns)). Randomised multicentre clinical trial. Post hoc analyses of the DASH-Sodium trial in the USA. In a multicentre feeding study with three 30 day periods, 390 participants were randomised to the DASH or control diet. On their assigned diet, participants ate food with high sodium during one period, intermediate sodium during another period and low sodium during another period, in random order. Occurrence and severity of headache were ascertained from self-administered questionnaires, completed at the end of each feeding period. The occurrence of headaches was similar in DASH versus control, at high (OR (95% CI)=0.65 (0.37 to 1.12); p=0.12), intermediate (0.57 (0.29 to 1.12); p=0.10) and low (0.64 (0.36 to 1.13); p=0.12) sodium levels. By contrast, there was a lower risk of headache on the low, compared with high, sodium level, both on the control (0.69 (0.49 to 0.99); p=0.05) and DASH (0.69 (0.49 to 0.98); p=0.04) diets. A reduced sodium intake was associated with a significantly lower risk of headache, while dietary patterns had no effect on the risk of headaches in adults. Reduced dietary sodium intake offers a novel approach to prevent headaches. NCT00000608. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Effects of dietary sodium and the DASH diet on the occurrence of headaches: results from randomised multicentre DASH-Sodium clinical trial

PubMed Central

Amer, Muhammad; Woodward, Mark; Appel, Lawrence J

2014-01-01

Objectives Headaches are a common medical problem, yet few studies, particularly trials, have evaluated therapies that might prevent or control headaches. We, thus, investigated the effects on the occurrence of headaches of three levels of dietary sodium intake and two diet patterns (the Dietary Approaches to Stop Hypertension (DASH) diet (rich in fruits, vegetables and low-fat dairy products with reduced saturated and total fat) and a control diet (typical of Western consumption patterns)). Design Randomised multicentre clinical trial. Setting Post hoc analyses of the DASH-Sodium trial in the USA. Participants In a multicentre feeding study with three 30 day periods, 390 participants were randomised to the DASH or control diet. On their assigned diet, participants ate food with high sodium during one period, intermediate sodium during another period and low sodium during another period, in random order. Outcome measures Occurrence and severity of headache were ascertained from self-administered questionnaires, completed at the end of each feeding period. Results The occurrence of headaches was similar in DASH versus control, at high (OR (95% CI)=0.65 (0.37 to 1.12); p=0.12), intermediate (0.57 (0.29 to 1.12); p=0.10) and low (0.64 (0.36 to 1.13); p=0.12) sodium levels. By contrast, there was a lower risk of headache on the low, compared with high, sodium level, both on the control (0.69 (0.49 to 0.99); p=0.05) and DASH (0.69 (0.49 to 0.98); p=0.04) diets. Conclusions A reduced sodium intake was associated with a significantly lower risk of headache, while dietary patterns had no effect on the risk of headaches in adults. Reduced dietary sodium intake offers a novel approach to prevent headaches. Trial registration number NCT00000608. PMID:25500372

Adherence to Dietary Approaches to Stop Hypertension (DASH) Dietary Pattern in Relation to Chronic Obstructive Pulmonary Disease (COPD): A Case-Control Study.

PubMed

Ardestani, Mohammad Emami; Onvani, Shokouh; Esmailzadeh, Ahmad; Feizi, Awat; Azadbakht, Leila

2017-01-01

This case-control study was designed to investigate the association between adherences to the Dietary Approaches to Stop Hypertension (DASH) diet in patients with chronic obstructive pulmonary disease (COPD) in comparison to subjects without COPD. This is a case-control study. Usual dietary intake was assessed using a validated food frequency questionnaire. Lung function was evaluated with spirometry testing, and one of the researchers inquired about other respiratory symptoms, including chronic cough, sputum, and breathlessness. Adherence to the DASH dietary pattern was assessed according to the Fung method. This study was conducted at Alzahra University Hospital of Isfahan, Iran, in 2015. Eight-four patients with COPD and 80 subjects without a history of COPD participated in study. The mean age of participants was 57 years. Average smoking in the case group was about 27.5 pack-years. Spirometry tests including forced expiratory volume in 1 second (FEV 1 ), forced vital capacity (FVC), and FEV 1 /FVC were significantly lower in patients with COPD (p = 0.0001). Among COPD symptoms, cough was significantly decreased across tertiles of DASH score (p = 0.03). Significant differences were found for DASH score between patients with COPD and control subjects (19.82 + 3.63 vs 21.13 + 3.82, p = 0.02). Vitamin C, vitamin E, and dietary fiber intake were lower in patients with COPD (144.32 + 70.51 vs 166.97 + 71.88, p = 0.04, 7.49 + 3.91 vs 8.72 + 3.21, p = 0.02 and 19.34 + 7.05 vs 22.19 + 7.87, p = 0.01, respectively). We observed that adherence to a DASH dietary pattern among patients with COPD was significantly lower compared to the control group. Cough was significantly decreased by increments in adherence to a DASH dietary pattern.

Alveolar Macrophages Play a Key Role in Cockroach-Induced Allergic Inflammation via TNF-α Pathway

PubMed Central

Kim, Joo Young; Sohn, Jung Ho; Choi, Je-Min; Lee, Jae-Hyun; Hong, Chein-Soo; Lee, Joo-Shil; Park, Jung-Won

2012-01-01

The activity of the serine protease in the German cockroach allergen is important to the development of allergic disease. The protease-activated receptor (PAR)-2, which is expressed in numerous cell types in lung tissue, is known to mediate the cellular events caused by inhaled serine protease. Alveolar macrophages express PAR-2 and produce considerable amounts of tumor necrosis factor (TNF)-α. We determined whether the serine protease in German cockroach extract (GCE) enhances TNF-α production by alveolar macrophages through the PAR-2 pathway and whether the TNF-α production affects GCE-induced pulmonary inflammation. Effects of GCE on alveolar macrophages and TNF-α production were evaluated using in vitro MH-S and RAW264.6 cells and in vivo GCE-induced asthma models of BALB/c mice. GCE contained a large amount of serine protease. In the MH-S and RAW264.7 cells, GCE activated PAR-2 and thereby produced TNF-α. In the GCE-induced asthma model, intranasal administration of GCE increased airway hyperresponsiveness (AHR), inflammatory cell infiltration, productions of serum immunoglobulin E, interleukin (IL)-5, IL-13 and TNF-α production in alveolar macrophages. Blockade of serine proteases prevented the development of GCE induced allergic pathologies. TNF-α blockade also prevented the development of such asthma-like lesions. Depletion of alveolar macrophages reduced AHR and intracellular TNF-α level in pulmonary cell populations in the GCE-induced asthma model. These results suggest that serine protease from GCE affects asthma through an alveolar macrophage and TNF-α dependent manner, reflecting the close relation of innate and adaptive immune response in allergic asthma model. PMID:23094102

m-AAA proteases, mitochondrial calcium homeostasis and neurodegeneration

PubMed Central

Patron, Maria; Sprenger, Hans-Georg; Langer, Thomas

2018-01-01

The function of mitochondria depends on ubiquitously expressed and evolutionary conserved m-AAA proteases in the inner membrane. These ATP-dependent peptidases form hexameric complexes built up of homologous subunits. AFG3L2 subunits assemble either into homo-oligomeric isoenzymes or with SPG7 (paraplegin) subunits into hetero-oligomeric proteolytic complexes. Mutations in AFG3L2 are associated with dominant spinocerebellar ataxia (SCA28) characterized by the loss of Purkinje cells, whereas mutations in SPG7 cause a recessive form of hereditary spastic paraplegia (HSP7) with motor neurons of the cortico-spinal tract being predominantly affected. Pleiotropic functions have been assigned to m-AAA proteases, which act as quality control and regulatory enzymes in mitochondria. Loss of m-AAA proteases affects mitochondrial protein synthesis and respiration and leads to mitochondrial fragmentation and deficiencies in the axonal transport of mitochondria. Moreover m-AAA proteases regulate the assembly of the mitochondrial calcium uniporter (MCU) complex. Impaired degradation of the MCU subunit EMRE in AFG3L2-deficient mitochondria results in the formation of deregulated MCU complexes, increased mitochondrial calcium uptake and increased vulnerability of neurons for calcium-induced cell death. A reduction of calcium influx into the cytosol of Purkinje cells rescues ataxia in an AFG3L2-deficient mouse model. In this review, we discuss the relationship between the m-AAA protease and mitochondrial calcium homeostasis and its relevance for neurodegeneration and describe a novel mouse model lacking MCU specifically in Purkinje cells. Our results pledge for a novel view on m-AAA proteases that integrates their pleiotropic functions in mitochondria to explain the pathogenesis of associated neurodegenerative disorders. PMID:29451229

m-AAA proteases, mitochondrial calcium homeostasis and neurodegeneration.

PubMed

Patron, Maria; Sprenger, Hans-Georg; Langer, Thomas

2018-03-01

The function of mitochondria depends on ubiquitously expressed and evolutionary conserved m-AAA proteases in the inner membrane. These ATP-dependent peptidases form hexameric complexes built up of homologous subunits. AFG3L2 subunits assemble either into homo-oligomeric isoenzymes or with SPG7 (paraplegin) subunits into hetero-oligomeric proteolytic complexes. Mutations in AFG3L2 are associated with dominant spinocerebellar ataxia (SCA28) characterized by the loss of Purkinje cells, whereas mutations in SPG7 cause a recessive form of hereditary spastic paraplegia (HSP7) with motor neurons of the cortico-spinal tract being predominantly affected. Pleiotropic functions have been assigned to m-AAA proteases, which act as quality control and regulatory enzymes in mitochondria. Loss of m-AAA proteases affects mitochondrial protein synthesis and respiration and leads to mitochondrial fragmentation and deficiencies in the axonal transport of mitochondria. Moreover m-AAA proteases regulate the assembly of the mitochondrial calcium uniporter (MCU) complex. Impaired degradation of the MCU subunit EMRE in AFG3L2-deficient mitochondria results in the formation of deregulated MCU complexes, increased mitochondrial calcium uptake and increased vulnerability of neurons for calcium-induced cell death. A reduction of calcium influx into the cytosol of Purkinje cells rescues ataxia in an AFG3L2-deficient mouse model. In this review, we discuss the relationship between the m-AAA protease and mitochondrial calcium homeostasis and its relevance for neurodegeneration and describe a novel mouse model lacking MCU specifically in Purkinje cells. Our results pledge for a novel view on m-AAA proteases that integrates their pleiotropic functions in mitochondria to explain the pathogenesis of associated neurodegenerative disorders.

Scalable privacy-preserving data sharing methodology for genome-wide association studies: an application to iDASH healthcare privacy protection challenge.

PubMed

Yu, Fei; Ji, Zhanglong

2014-01-01

In response to the growing interest in genome-wide association study (GWAS) data privacy, the Integrating Data for Analysis, Anonymization and SHaring (iDASH) center organized the iDASH Healthcare Privacy Protection Challenge, with the aim of investigating the effectiveness of applying privacy-preserving methodologies to human genetic data. This paper is based on a submission to the iDASH Healthcare Privacy Protection Challenge. We apply privacy-preserving methods that are adapted from Uhler et al. 2013 and Yu et al. 2014 to the challenge's data and analyze the data utility after the data are perturbed by the privacy-preserving methods. Major contributions of this paper include new interpretation of the χ2 statistic in a GWAS setting and new results about the Hamming distance score, a key component for one of the privacy-preserving methods.

Scalable privacy-preserving data sharing methodology for genome-wide association studies: an application to iDASH healthcare privacy protection challenge

PubMed Central

2014-01-01

In response to the growing interest in genome-wide association study (GWAS) data privacy, the Integrating Data for Analysis, Anonymization and SHaring (iDASH) center organized the iDASH Healthcare Privacy Protection Challenge, with the aim of investigating the effectiveness of applying privacy-preserving methodologies to human genetic data. This paper is based on a submission to the iDASH Healthcare Privacy Protection Challenge. We apply privacy-preserving methods that are adapted from Uhler et al. 2013 and Yu et al. 2014 to the challenge's data and analyze the data utility after the data are perturbed by the privacy-preserving methods. Major contributions of this paper include new interpretation of the χ2 statistic in a GWAS setting and new results about the Hamming distance score, a key component for one of the privacy-preserving methods. PMID:25521367

Capture of SO3 isomers in the oxidation of sulfur monoxide with molecular oxygen.

PubMed

Wu, Zhuang; Lu, Bo; Feng, Ruijuan; Xu, Jian; Lu, Yan; Wan, Huabin; Eckhardt, André K; Schreiner, Peter R; Xie, Changjian; Guo, Hua; Zeng, Xiaoqing

2018-02-13

When mixing SO with O 2 in N 2 , Ne, or Ar, an end-on complex OS-OO forms in the gas phase and can subsequently be trapped at cryogenic temperatures (2.8-15.0 K). Upon infrared light irradiation, OS-OO converts to SO 3 and SO 2 + O with the concomitant formation of a rare 1,2,3-dioxathiirane 2-oxide, i.e., cyclic OS([double bond, length as m-dash]O)O. Unexpectedly, the ring-closure of 16 OS- 18 O 18 O yields a ca. 2 : 1 mixture of cyclic 18 OS([double bond, length as m-dash] 16 O) 18 O and 16 OS([double bond, length as m-dash] 18 O) 18 O. The characterization of OS-OO and OS([double bond, length as m-dash]O)O with IR and UV/Vis spectroscopy is supported by high-level ab initio computations.

Tailoring the photoluminescence polarization anisotropy of a single InAs quantum dash by a post-growth modification of its dielectric environment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mrowiński, P.; Misiewicz, J.; Sęk, G.

Excitonic emission from single InAs/InGaAlAs/InP quantum dashes has been investigated in terms of controlling the polarization anisotropy by altering the shape of the processed sub-micrometer mesa structures. Photoluminescence has been measured from exemplary single quantum dashes emitting around 1.3 and 1.55 μm and placed inside rectangular mesas of various orientation, asymmetry, and sizes. The detected degree of linear polarization of bright exciton emission ranges from −0.1 to ca. 0.55, compared to 0.25 for dashes in unaltered or isotropic in-plane dielectric surrounding. These results are interpreted by numerical simulations using an emitter coupled with a single optical mode in such a mesamore » and outgoing in the direction normal to the sample surface.« less

Associations of diet quality with cognition in children - the Physical Activity and Nutrition in Children Study.

PubMed

Haapala, Eero A; Eloranta, Aino-Maija; Venäläinen, Taisa; Schwab, Ursula; Lindi, Virpi; Lakka, Timo A

2015-10-14

Evidence on the associations of dietary patterns with cognition in children is limited. Therefore, we investigated the associations of the Baltic Sea Diet Score (BSDS) and the Dietary Approaches to Stop Hypertension (DASH) score with cognition in children. The present cross-sectional study sample included 428 children aged 6-8 years (216 boys and 212 girls). The BSDS and the DASH score were calculated using data from 4 d food records, higher scores indicating better diet quality. Cognition was assessed by the Raven's Coloured Progressive Matrices (CPM) score, a higher score indicating better cognition. Among all children, the BSDS (standardised regression coefficient β = 0·122, P =0·012) and the DASH score (β = 0·121, P =0·015) were directly associated with the Raven's CPM score. Among boys, a lower BSDS (β = 0·244, P< 0·001) and a lower DASH score (β = 0·202, P= 0·003) were related to a lower Raven's CPM score. Boys in the lowest quartile of the BSDS (22·5 v. 25·3, P= 0·029) and the DASH score (22·4 v. 25·7, P= 0·008) had a lower Raven's CPM score than those in the highest quartile of the corresponding score. Among girls, the BSDS or the DASH score were not associated with cognition. In conclusion, a poorer diet quality was associated with worse cognition in children, and the relationship was stronger in boys than in girls.

Construct validity of the canadian occupational performance measure in participants with tendon injury and Dupuytren disease.

PubMed

van de Ven-Stevens, Lucelle A W; Graff, Maud J L; Peters, Marlijn A M; van der Linde, Harmen; Geurts, Alexander C H

2015-05-01

In patient-centered practice, instruments need to assess outcomes that are meaningful to patients with hand conditions. It is unclear which assessment tools address these subjective perspectives best. The aim of this study was to establish the construct validity of the Canadian Occupational Performance Measure (COPM) in relation to the Disabilities of Arm, Shoulder, and Hand (DASH) questionnaire and the Michigan Hand Outcomes Questionnaire (MHQ) in people with hand conditions. It was hypothesized that COPM scores would correlate with DASH and MHQ total scores only to a moderate degree and that the COPM, DASH questionnaire, and MHQ would all correlate weakly with measures of hand impairments. This was a validation study. The COPM, DASH questionnaire, and MHQ were scored, and then hand impairments were measured (pain [numerical rating scale], active range of motion [goniometer], grip strength [dynamometer], and pinch grip strength [pinch meter]). People who had received postsurgery rehabilitation for flexor tendon injuries, extensor tendon injuries, or Dupuytren disease were eligible. Seventy-two participants were included. For all diagnosis groups, the Pearson coefficient of correlation between the DASH questionnaire and the MHQ was higher than .60, whereas the correlation between the performance scale of the COPM and either the DASH questionnaire or the MHQ was lower than .51. Correlations of these assessment tools with measures of hand impairments were lower than .46. The small sample sizes may limit the generalization of the results. The results supported the hypotheses and, thus, the construct validity of the COPM after surgery in people with hand conditions. © 2015 American Physical Therapy Association.

Effects of Different Dietary Interventions on Calcitriol, Parathyroid Hormone, Calcium, and Phosphorus: Results from the DASH Trial

PubMed Central

Michos, Erin D.; Miller, Edgar R.; Crisp, Zeni

2018-01-01

The “Dietary Approaches to Stop Hypertension” (DASH) diet, rich in fiber and low-fat dairy, effectively lowers blood pressure. DASH’s effect on calcitriol and other markers of bone-mineral metabolism is unknown. This secondary analysis of the DASH trial aimed to determine the effect of dietary patterns on blood concentrations of calcitriol, parathyroid hormone (PTH), ionized calcium, and urinary excretion of calcium and phosphorus. Outcomes were available in 334 participants in the trial. After a 3-week run-in on the control diet, participants were randomized to control, fruits and vegetables (F&V), or DASH diets. Outcomes were assessed at the end of run-in, and during the last week of the intervention period. Mean age of participants was 45.7 ± 10.7 years, 46% female, and 57% African-American. Mean ± Standard Deviation(SD) baseline serum concentrations of calcitriol, PTH, and ionized calcium were 37.8 ± 9.2 pg/mL, 46.1 ± 18.5 pg/mL and 5.2 ± 0.23 mg/dL, respectively. Mean (±SD) urinary calcium and phosphorus excretions were 150.1 ± 77.8 and 708.0 ± 251.8 mg/24 h, respectively. Compared with control, DASH reduced calcitriol −3.32 pg/mL (p = 0.004). Otherwise, there was no significant effect on other biomarkers. DASH lowered serum calcitriol perhaps more among African-Americans. These results raise important questions about the interpretation and clinical significance of low calcitriol concentrations in the setting of recommended diets. PMID:29562597

A novel serine protease from strawberry (Fragaria ananassa): Purification and biochemical characterization.

PubMed

Alici, Esma Hande; Arabaci, Gulnur

2018-03-27

In this study, a protease enzyme was purified from strawberry by using Sepharose-4B-l-tyrosine-p-amino benzoic acid affinity chromatography. The molecular weight of pure protease was determined 65.8 kDa by SDS-PAGE. The single band observed on the gel showed that the enzyme had a single polypeptide chain and was successfully purified. Purification of the protease by the chromatographic method resulted in a 395.6-fold increase in specific activity (3600 U/mg). Optimum pH and temperature for the enzyme were 6 and 40 °C, respectively. The protease was stable at a wide temperature range of 40 to 70 °C and a pH range of 3.0 to 9.0. Co 2+ ions stimulated protease activity very strongly. Cu 2+ , Hg 2+ , Cd 2+ and Mn 2+ ions significantly inhibited protease activity. While 2-propanol completely inhibited the enzyme, the enzyme maintained its activity better in the presence of ethanol and methanol. The strawberry protease showed the highest specificity towards hemoglobin among all the natural substrates tested. The specificity of the enzyme towards synthetic substrates was also investigated and it was concluded that it has broad substrate specificity. The obtained results indicated that this purified protease was highly-likely a serine protease and its activity was significantly affected by the presence of metal ions. Copyright © 2018. Published by Elsevier B.V.

Activity of the Human Rhinovirus 3C Protease Studied in Various Buffers, Additives and Detergents Solutions for Recombinant Protein Production

PubMed Central

Tufail, Soban; Ismat, Fouzia; Imran, Muhammad; Iqbal, Mazhar; Mirza, Osman; Rhaman, Moazur

2016-01-01

Proteases are widely used to remove affinity and solubility tags from recombinant proteins to avoid potential interference of these tags with the structure and function of the fusion partner. In recent years, great interest has been seen in use of the human rhinovirus 3C protease owing to its stringent sequence specificity and enhanced activity. Like other proteases, activity of the human rhinovirus 3C protease can be affected in part by the buffer components and additives that are generally employed for purification and stabilization of proteins, hence, necessitate their removal by tedious and time-consuming procedures before proteolysis can occur. To address this issue, we examined the effect of elution buffers used for common affinity based purifications, salt ions, stability/solubility and reducing agents, and detergents on the activity of the human rhinovirus 3C protease using three different fusion proteins at 4°C, a temperature of choice for purification of many proteins. The results show that the human rhinovirus 3C protease performs better at 4°C than the frequently used tobacco etch virus protease and its activity was insensitive to most of the experimental conditions tested. Though number of fusion proteins tested is limited, we expect that these finding will facilitate the use of the human rhinovirus 3C protease in recombinant protein production for pharmaceutical and biotechnological applications. PMID:27093053

Activity of the Human Rhinovirus 3C Protease Studied in Various Buffers, Additives and Detergents Solutions for Recombinant Protein Production.

PubMed

Ullah, Raheem; Shah, Majid Ali; Tufail, Soban; Ismat, Fouzia; Imran, Muhammad; Iqbal, Mazhar; Mirza, Osman; Rhaman, Moazur

2016-01-01

Proteases are widely used to remove affinity and solubility tags from recombinant proteins to avoid potential interference of these tags with the structure and function of the fusion partner. In recent years, great interest has been seen in use of the human rhinovirus 3C protease owing to its stringent sequence specificity and enhanced activity. Like other proteases, activity of the human rhinovirus 3C protease can be affected in part by the buffer components and additives that are generally employed for purification and stabilization of proteins, hence, necessitate their removal by tedious and time-consuming procedures before proteolysis can occur. To address this issue, we examined the effect of elution buffers used for common affinity based purifications, salt ions, stability/solubility and reducing agents, and detergents on the activity of the human rhinovirus 3C protease using three different fusion proteins at 4°C, a temperature of choice for purification of many proteins. The results show that the human rhinovirus 3C protease performs better at 4°C than the frequently used tobacco etch virus protease and its activity was insensitive to most of the experimental conditions tested. Though number of fusion proteins tested is limited, we expect that these finding will facilitate the use of the human rhinovirus 3C protease in recombinant protein production for pharmaceutical and biotechnological applications.

Dots & dashes : piecing together transit's future.

DOT National Transportation Integrated Search

2008-11-01

Dots & Dashes is an interactive board game that is used at public planning meetings to engage stakeholders in long range transit planning. Groups of three to six people use game pieces with monetary values to choose the priorities of transit projects...

FleetDASH

DOE Office of Scientific and Technical Information (OSTI.GOV)

Singer, Mark R

2017-09-06

FleetDASH helps federal fleet managers maximize their use of alternative fuel. This presentation explains how the dashboard works and demonstrates the newest capabilities added to the tool. It also reviews complementary online tools available to fleet managers on the Alternative Fuel Data Center.

A Collection of Technical Studies Completed for the Computer-Aided Acquisition and Logistic Support (CALS) Program Fiscal Year 1988. Volume 3. CGM Registration

DTIC Science & Technology

1991-03-01

the array are used cyclically, that is when the end of the array is reached, the pattern starts over at the beginning. Dashed lines wrap around curves...the dash pattern relative to the start of the path. It is interpreted as a .distance into the dash pattern at which the pattern should be started ...cubic seldom Is drawn using the four points specified. The curve starts at the first point and ends at the fourth point; the second and third point are

Postoperative improvement in DASH score, clinical findings, and nerve conduction velocity in patients with cubital tunnel syndrome.

PubMed

Ido, Yoshikazu; Uchiyama, Shigeharu; Nakamura, Koichi; Itsubo, Toshiro; Hayashi, Masanori; Hata, Yukihiko; Imaeda, Toshihiko; Kato, Hiroyuki

2016-06-06

We investigated a recovery pattern in subjective and objective measures among 52 patients with cubital tunnel syndrome after anterior subcutaneous transposition of the ulnar nerve. Disabilities of the Arm, Shoulder and Hand (DASH) score (primary outcome), numbness score, grip and pinch strength, Semmes-Weinstein (SW) score, static 2-point discrimination (2PD) score, and motor conduction velocity (MCV) stage were examined preoperatively and 1, 3, 6, 12, and ≥24 months postoperatively. Statistical analyses were conducted to evaluate how each variable improved after surgery. A linear mixed-effects model was used for continuous variables (DASH score, numbness, grip and pinch strength), and a proportional odds model was used for categorical variables (SW and 2PD tests and MCV stages). DASH score significantly improved by 6 months. Significant recovery in numbness and SW test scores occurred at 1 month. Grip and pinch strength, 2PD test scores, and MCV stage improved by 3 months. DASH scores and numbness recovered regardless of age, sex, or disease severity. It was still unclear if both subjective and objective measures improved beyond 1-year postoperatively. These data are helpful for predicting postoperative recovery patterns and tend to be most important for patients prior to surgery.

Effect of Culture Conditions on the Production of an Extracellular Protease by Bacillus sp. Isolated from Soil Sample of Lavizan Jungle Park

PubMed Central

Akhavan Sepahy, Abbas; Jabalameli, Leila

2011-01-01

Soil samples of Tehran jungle parks were screened for proteolytic Bacilli. Among eighteen protease producers one of the isolates obtained from Lavizan park, in north east of Tehran, was selected for further experimental studies. This isolate was identified as Bacillus sp. strain CR-179 based on partial sequencing of 16S rRNA. Various nutritional and environmental parameters affected protease production by Bacillus sp. strain CR-179. Protease production by this Bacillus cultivated in liquid cultures reached a maximum at 24 h, with levels of 340.908 U/mL. Starch and maltose were the best substrates for enzyme production while some pure sugars such as fructose, glucose, and sucrose could not influence production of protease. Among various organic nitrogen sources corn steep liquor, which is commercial, was found as the best substrate followed by yeast extract, whey protein, and beef extract. The optimal pH and optimal temperature of enzyme production were 8.0 and 45°C, respectively. Studies on enzymatic characterization revealed that crude protease showed maximum activity at pH 9.0 and 60°C, which is indicating the enzyme to be thermoalkaline protease. PMID:22191016

Characterization of molecular interactions between Zika virus protease and peptides derived from the C-terminus of NS2B.

PubMed

Li, Yan; Loh, Ying Ru; Hung, Alvin W; Kang, CongBao

2018-06-21

Zika virus (ZIKV) protease is a two-component complex in which NS3 contains the catalytic triad and NS2B cofactor region is important for protease folding and activity. A protease construct-eZiPro without the transmembrane domains of NS2B was designed. Structural study on eZiPro reveals that the Thr-Gly-Lys-Arg (TGKR) sequence at the C-terminus of NS2B binds to the active site after cleavage. The bZiPro construct only contains NS2B cofactor region and the N-terminus of NS3 without any artificial linker or protease cleavage site, giving rise to an empty pocket accessible to substrate and inhibitor binding. Herein, we demonstrate that the TGKR sequence of NS2B in eZiPro is dynamic. Peptides from NS2B with various lengths exhibit different binding affinities to bZiPro. TGKR binding to the active site in eZiPro does not affect protease binding to small-molecule compounds. Our results suggest that eZiPro will also be useful for evaluating small-molecule protease inhibitors. Copyright © 2018 Elsevier Inc. All rights reserved.

Cysteine proteases and wheat (Triticum aestivum L) under drought: A still greatly unexplored association.

PubMed

Botha, Anna-Maria; Kunert, Karl J; Cullis, Christopher A

2017-09-01

Bread wheat (Triticum aestivum L.) provides about 19% of global dietary energy. Environmental stress, such as drought, affects wheat growth causing premature plant senescence and ultimately plant death. A plant response to drought is an increase in protease-mediated proteolysis with rapid degradation of proteins required for metabolic processes. Among the plant proteases that are increased in their activity following stress, cysteine proteases are the best characterized. Very little is known about particular wheat cysteine protease sequences, their expression and also localization. The current knowledge on wheat cysteine proteases belonging to the five clans (CA, CD, CE, CF and CP) is outlined, in particular their expression and possible function under drought. The first successes in establishing an annotated wheat genome database are further highlighted which has allowed more detailed mining of cysteine proteases. We also share our thoughts on future research directions considering the growing availability of genomic resources of this very important food crop. Finally, we also outline future application of developed knowledge in transgenic wheat plants for environmental stress protection and also as senescence markers to monitor wheat growth under environmental stress conditions. © 2017 John Wiley & Sons Ltd.

Protease-Mediated Suppression of DRG Neuron Excitability by Commensal Bacteria.

PubMed

Sessenwein, Jessica L; Baker, Corey C; Pradhananga, Sabindra; Maitland, Megan E; Petrof, Elaine O; Allen-Vercoe, Emma; Noordhof, Curtis; Reed, David E; Vanner, Stephen J; Lomax, Alan E

2017-11-29

Peripheral pain signaling reflects a balance of pronociceptive and antinociceptive influences; the contribution by the gastrointestinal microbiota to this balance has received little attention. Disorders, such as inflammatory bowel disease and irritable bowel syndrome, are associated with exaggerated visceral nociceptive actions that may involve altered microbial signaling, particularly given the evidence for bacterial dysbiosis. Thus, we tested whether a community of commensal gastrointestinal bacteria derived from a healthy human donor (microbial ecosystem therapeutics; MET-1) can affect the excitability of male mouse DRG neurons. MET-1 reduced the excitability of DRG neurons by significantly increasing rheobase, decreasing responses to capsaicin (2 μm) and reducing action potential discharge from colonic afferent nerves. The increase in rheobase was accompanied by an increase in the amplitude of voltage-gated K + currents. A mixture of bacterial protease inhibitors abrogated the effect of MET-1 effects on DRG neuron rheobase. A serine protease inhibitor but not inhibitors of cysteine proteases, acid proteases, metalloproteases, or aminopeptidases abolished the effects of MET-1. The serine protease cathepsin G recapitulated the effects of MET-1 on DRG neurons. Inhibition of protease-activated receptor-4 (PAR-4), but not PAR-2, blocked the effects of MET-1. Furthermore, Faecalibacterium prausnitzii recapitulated the effects of MET-1 on excitability of DRG neurons. We conclude that serine proteases derived from commensal bacteria can directly impact the excitability of DRG neurons, through PAR-4 activation. The ability of microbiota-neuronal interactions to modulate afferent signaling suggests that therapies that induce or correct microbial dysbiosis may impact visceral pain. SIGNIFICANCE STATEMENT Commercially available probiotics have the potential to modify visceral pain. Here we show that secretory products from gastrointestinal microbiota derived from a human donor signal to DRG neurons. Their secretory products contain serine proteases that suppress excitability via activation of protease-activated receptor-4. Moreover, from this community of commensal microbes, Faecalibacterium prausnitzii strain 16-6-I 40 fastidious anaerobe agar had the greatest effect. Our study suggests that therapies that induce or correct microbial dysbiosis may affect the excitability of primary afferent neurons, many of which are nociceptive. Furthermore, identification of the bacterial strains capable of suppressing sensory neuron excitability, and their mechanisms of action, may allow therapeutic relief for patients with gastrointestinal diseases associated with pain. Copyright © 2017 the authors 0270-6474/17/3711758-11$15.00/0.

Clinical rating systems in elbow research-a systematic review exploring trends and distributions of use.

PubMed

Evans, Jonathan P; Smith, Chris D; Fine, Nicola F; Porter, Ian; Gangannagaripalli, Jaheeda; Goodwin, Victoria A; Valderas, Jose M

2018-04-01

Clinical rating systems are used as outcome measures in clinical trials and attempt to gauge the patient's view of his or her own health. The choice of clinical rating system should be supported by its performance against established quality standards. A search strategy was developed to identify all studies that reported the use of clinical rating systems in the elbow literature. The strategy was run from inception in Medline Embase and CINHAL. Data extraction identified the date of publication, country of data collection, pathology assessed, and the outcome measure used. We identified 980 studies that reported clinical rating system use. Seventy-two separate rating systems were identified. Forty-one percent of studies used ≥2 separate measures. Overall, 54% of studies used the Mayo Elbow Performance Score (MEPS). For arthroplasty, 82% used MEPS, 17% used Disabilities of Arm, Shoulder and Hand (DASH), and 7% used QuickDASH. For trauma, 66.7% used MEPS, 32% used DASH, and 23% used the Morrey Score. For tendinopathy, 31% used DASH, 23% used Patient-Rated Tennis Elbow Evaluation (PRTEE), and 13% used MEPS. Over time, there was an increased proportional use of the MEPS, DASH, QuickDASH, PRTEE, and the Oxford Elbow Score. This study identified a wide choice and usage of clinical rating systems in the elbow literature. Numerous studies reported measures without a history of either a specific pathology or cross-cultural validation. Interpretability and comparison of outcomes is dependent on the unification of outcome measure choice. This was not demonstrated currently. Copyright © 2018 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.

Association between adherence to the Dietary Approaches to Stop Hypertension diet with food security and weight status in adult women.

PubMed

Tabibian, Saeideh; Daneshzad, Elnaz; Bellissimo, Nick; Brett, Neil R; Dorosty-Motlagh, Ahmad R; Azadbakht, Leila

2018-06-10

To investigate the association between adherence to the Dietary Approaches to Stop Hypertension (DASH) diet with food security and weight status in adult women. This cross-sectional study was carried out in 227 women-20-50 years of age-who were referred from 10 health centres. Dietary intakes were assessed using validated food frequency questionnaires. The DASH score was calculated using the Fung method based on eight food and nutrient components (high intake of fruits, vegetables, whole grains, nuts and legumes, low-fat dairy, low intakes of red and processed meats, sweetened beverages and sodium). The United States Department of Agriculture (USDA) household food security questionnaire was used to assess food security status. The prevalence of food insecurity was 33.9%. A greater percentage of overweight and obese people were in the food insecure group (P = 0.006). In addition, the prevalence of overweight and obese people was lower with greater adherence to the DASH dietary pattern (P = 0.017). After controlling for age and energy intake, participants in the highest tertile of adherence to DASH diet had 66% lower odds of overweight and obesity than those in the lowest tertile (odds ratio (OR): 0.34; 95% confidence interval (CI): 0.15; 0.79). This relationship of DASH diet tertile and overweight and obesity was significant for both food secure women (OR: 0.54, 95% CI: 0.23; 0.97) and food insecure women (OR: 0.35, 95% CI: 0.06; 0.42). Adherence to the DASH diet is associated with a reduced risk of overweight and obesity, based on body mass index, in both food secure and insecure Iranian women. © 2018 Dietitians Association of Australia.

Effects of the dietary approaches to stop hypertension diet, exercise, and caloric restriction on neurocognition in overweight adults with high blood pressure.

PubMed

Smith, Patrick J; Blumenthal, James A; Babyak, Michael A; Craighead, Linda; Welsh-Bohmer, Kathleen A; Browndyke, Jeffrey N; Strauman, Timothy A; Sherwood, Andrew

2010-06-01

High blood pressure increases the risks of stroke, dementia, and neurocognitive dysfunction. Although aerobic exercise and dietary modifications have been shown to reduce blood pressure, no randomized trials have examined the effects of aerobic exercise combined with dietary modification on neurocognitive functioning in individuals with high blood pressure (ie, prehypertension and stage 1 hypertension). As part of a larger investigation, 124 participants with elevated blood pressure (systolic blood pressure 130 to 159 mm Hg or diastolic blood pressure 85 to 99 mm Hg) who were sedentary and overweight or obese (body mass index: 25 to 40 kg/m(2)) were randomized to the Dietary Approaches to Stop Hypertension (DASH) diet alone, DASH combined with a behavioral weight management program including exercise and caloric restriction, or a usual diet control group. Participants completed a battery of neurocognitive tests of executive function-memory-learning and psychomotor speed at baseline and again after the 4-month intervention. Participants on the DASH diet combined with a behavioral weight management program exhibited greater improvements in executive function-memory-learning (Cohen's D=0.562; P=0.008) and psychomotor speed (Cohen's D=0.480; P=0.023), and DASH diet alone participants exhibited better psychomotor speed (Cohen's D=0.440; P=0.036) compared with the usual diet control. Neurocognitive improvements appeared to be mediated by increased aerobic fitness and weight loss. Also, participants with greater intima-medial thickness and higher systolic blood pressure showed greater improvements in executive function-memory-learning in the group on the DASH diet combined with a behavioral weight management program. In conclusion, combining aerobic exercise with the DASH diet and caloric restriction improves neurocognitive function among sedentary and overweight/obese individuals with prehypertension and hypertension.

Approaches for Analyzing the Roles of Mast Cells and Their Proteases In Vivo

PubMed Central

Galli, Stephen J.; Tsai, Mindy; Marichal, Thomas; Tchougounova, Elena; Reber, Laurent L.; Pejler, Gunnar

2016-01-01

The roles of mast cells in health and disease remain incompletely understood. While the evidence that mast cells are critical effector cells in IgE-dependent anaphylaxis and other acute IgE-mediated allergic reactions seems unassailable, studies employing various mice deficient in mast cells or mast cell-associated proteases have yielded divergent conclusions about the roles of mast cells or their proteases in certain other immunological responses. Such “controversial” results call into question the relative utility of various older versus newer approaches to ascertain the roles of mast cells and mast cell proteases in vivo. This review discusses how both older and more recent mouse models have been used to investigate the functions of mast cells and their proteases in health and disease. We particularly focus on settings in which divergent conclusions about the importance of mast cells and their proteases have been supported by studies that employed different models of mast cell or mast cell protease deficiency. We think that two major conclusions can be drawn from such findings: (1) no matter which models of mast cell or mast cell protease deficiency one employs, the conclusions drawn from the experiments always should take into account the potential limitations of the models (particularly abnormalities affecting cell types other than mast cells) and (2) even when analyzing a biological response using a single model of mast cell or mast cell protease deficiency, details of experimental design are critical in efforts to define those conditions under which important contributions of mast cells or their proteases can be identified. PMID:25727288

47 CFR 2.926 - FCC identifier.

Code of Federal Regulations, 2011 CFR

2011-10-01

... Arabic numerals, capital letters, or combination thereof. A prospective grantee or his authorized... shall consist of a series of Arabic numerals, capital letters or a combination thereof, and may include the dash or hyphen (-). The total of Arabic numerals, capital letters and dashes or hyphens shall not...

47 CFR 2.926 - FCC identifier.

Code of Federal Regulations, 2010 CFR

2010-10-01

... Arabic numerals, capital letters, or combination thereof. A prospective grantee or his authorized... shall consist of a series of Arabic numerals, capital letters or a combination thereof, and may include the dash or hyphen (-). The total of Arabic numerals, capital letters and dashes or hyphens shall not...

Effectiveness of robot-assisted training added to conventional rehabilitation in patients with humeral fracture early after surgical treatment: protocol of a randomised, controlled, multicentre trial.

PubMed

Nerz, Corinna; Schwickert, Lars; Becker, Clemens; Studier-Fischer, Stefan; Müßig, Janina Anna; Augat, Peter

2017-12-06

The incidence of proximal humeral fractures increases with age. The functional recovery of the upper arm after such fractures is slow, and results are often disappointing. Treatment is associated with long immobilisation periods. Evidence-based exercise guidelines are missing. Loss of muscle mass as well as reduced range of motion and motor performance are common consequences. These losses could be partly counteracted by training interventions using robot-assisted arm support of the affected arm derived from neurorehabilitation. Thus, shorter immobilisation could be reached. Thus far, this approach has been tested in only a few small studies. The aim of the present study is to examine whether assistive robotic training augmenting conventional occupational and physical therapy can improve functional shoulder outcomes. Patients aged between 35 and 66 years with proximal humeral fracture and surgical treatment will be recruited at three different clinics in Germany and randomised into an intervention group and a control group. Participants will be assessed before randomisation and followed after completing an intervention period of 3 weeks and additionally after 3, 6 and 12 months. The baseline assessment will include cognition (Short Orientation-Memory-Concentration Test); level of pain in the affected arm; ability to work; gait speed (10-m walk); disability of the arm, shoulder and hand (Disabilities of the Arm, Shoulder and Hand Outcome Measure [DASH]); range of motion of the affected arm (goniometer measurement); visual acuity; and motor function of orthopaedic patients (Wolf Motor Function Test-Orthopaedic version [WMFT-O]). Clinical follow-up directly after the intervention will include assessment of disability of the arm, shoulder and hand (DASH) as well as range of motion and motor function (WMFT-O). The primary outcome parameter will be the DASH, and the secondary outcome parameter will be the WMFT-O. The long-term results will be assessed prospectively by postal follow-up. All patients will receive conventional occupational and physical therapy. The intervention group will receive additional robot-assisted training using the Armeo®Spring robot for 3 weeks. This study protocol describes a phase II, randomised, controlled, single-blind, multicentre intervention study. The results will guide and possibly improve methods of rehabilitation after proximal humeral fracture. Clinicaltrials.gov, NCT03100201 . Registered on 28 March 2017.

A low molecular weight serine protease: Purification and characterization from Hippasa agelenoides (funnel web) spider venom gland extract.

PubMed

Devaraja, S; Nagaraju, S; Mahadeswaraswamy, Y H; Girish, K S; Kemparaju, K

2008-07-01

Despite the long history [Kaiser, E., 1956. Enzymatic activity of spider venoms. In: Buckley, E.E., Porges, N. (Eds.), Venoms. American Association for the Advancement of Science, Washington, DC, pp. 91-93] on proteolytic activity, no study so far claims the isolation of a serine protease from the spider venom/venom gland extract. Therefore, the present study describes the isolation and characterization of a low molecular weight serine protease from Hippasa agelenoides venom gland extract. The protease (Hag-protease) was purified to homogeneity using the combination of gel-permeation and ion-exchange chromatography. The molecular mass was found to be 16.350 kDa by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. Hag-protease was optimally active at pH 7.5 and temperature of 37 degrees C. PMSF abolished the enzyme activity while EDTA, EGTA, IAA, 1, 10-phenanthrolene did not. It hydrolyzed proteins such as casein, fibronectin and collagen type-I dose dependently but did not degrade gelatin and collagen type-IV. The isolated protease was non-lethal and devoid of hemorrhagic, myotoxic and edema forming activities. The light microscopy of Hag-protease treated skin tissue sections at the site of injection showed extensive damage of extracellular matrix (ECM) of hypodermis without causing any damage to blood vessels and capillaries. Similar damage of ECM of muscle tissue sections without affecting myocytes was noticed. Hag-protease was found to be procoagulant in property when studied plasma recalcification time.

DASH-2: Flexible, Low-Cost, and High-Throughput SNP Genotyping by Dynamic Allele-Specific Hybridization on Membrane Arrays

PubMed Central

Jobs, Magnus; Howell, W. Mathias; Strömqvist, Linda; Mayr, Torsten; Brookes, Anthony J.

2003-01-01

Genotyping technologies need to be continually improved in terms of their flexibility, cost-efficiency, and throughput, to push forward genome variation analysis. To this end, we have leveraged the inherent simplicity of dynamic allele-specific hybridization (DASH) and coupled it to recent innovations of centrifugal arrays and iFRET. We have thereby created a new genotyping platform we term DASH-2, which we demonstrate and evaluate in this report. The system is highly flexible in many ways (any plate format, PCR multiplexing, serial and parallel array processing, spectral-multiplexing of hybridization probes), thus supporting a wide range of application scales and objectives. Precision is demonstrated to be in the range 99.8–100%, and assay costs are 0.05 USD or less per genotype assignment. DASH-2 thus provides a powerful new alternative for genotyping practice, which can be used without the need for expensive robotics support. PMID:12727908

iDASH: integrating data for analysis, anonymization, and sharing

PubMed Central

Bafna, Vineet; Boxwala, Aziz A; Chapman, Brian E; Chapman, Wendy W; Chaudhuri, Kamalika; Day, Michele E; Farcas, Claudiu; Heintzman, Nathaniel D; Jiang, Xiaoqian; Kim, Hyeoneui; Kim, Jihoon; Matheny, Michael E; Resnic, Frederic S; Vinterbo, Staal A

2011-01-01

iDASH (integrating data for analysis, anonymization, and sharing) is the newest National Center for Biomedical Computing funded by the NIH. It focuses on algorithms and tools for sharing data in a privacy-preserving manner. Foundational privacy technology research performed within iDASH is coupled with innovative engineering for collaborative tool development and data-sharing capabilities in a private Health Insurance Portability and Accountability Act (HIPAA)-certified cloud. Driving Biological Projects, which span different biological levels (from molecules to individuals to populations) and focus on various health conditions, help guide research and development within this Center. Furthermore, training and dissemination efforts connect the Center with its stakeholders and educate data owners and data consumers on how to share and use clinical and biological data. Through these various mechanisms, iDASH implements its goal of providing biomedical and behavioral researchers with access to data, software, and a high-performance computing environment, thus enabling them to generate and test new hypotheses. PMID:22081224

iDASH: integrating data for analysis, anonymization, and sharing.

PubMed

Ohno-Machado, Lucila; Bafna, Vineet; Boxwala, Aziz A; Chapman, Brian E; Chapman, Wendy W; Chaudhuri, Kamalika; Day, Michele E; Farcas, Claudiu; Heintzman, Nathaniel D; Jiang, Xiaoqian; Kim, Hyeoneui; Kim, Jihoon; Matheny, Michael E; Resnic, Frederic S; Vinterbo, Staal A

2012-01-01

iDASH (integrating data for analysis, anonymization, and sharing) is the newest National Center for Biomedical Computing funded by the NIH. It focuses on algorithms and tools for sharing data in a privacy-preserving manner. Foundational privacy technology research performed within iDASH is coupled with innovative engineering for collaborative tool development and data-sharing capabilities in a private Health Insurance Portability and Accountability Act (HIPAA)-certified cloud. Driving Biological Projects, which span different biological levels (from molecules to individuals to populations) and focus on various health conditions, help guide research and development within this Center. Furthermore, training and dissemination efforts connect the Center with its stakeholders and educate data owners and data consumers on how to share and use clinical and biological data. Through these various mechanisms, iDASH implements its goal of providing biomedical and behavioral researchers with access to data, software, and a high-performance computing environment, thus enabling them to generate and test new hypotheses.

Effects of Dietary Approach to Stop Hypertension diet on androgens, antioxidant status and body composition in overweight and obese women with polycystic ovary syndrome: a randomised controlled trial.

PubMed

Azadi-Yazdi, M; Karimi-Zarchi, M; Salehi-Abargouei, A; Fallahzadeh, H; Nadjarzadeh, A

2017-06-01

Polycystic ovary syndrome (PCOS) is the most common endocrine disease in reproductive age women. The present study aimed to determine the effects of Dietary Approaches to Stop Hypertension (DASH) diet on reproductive hormones, plasma total antioxidant status and anthropometric indices in overweight and obese PCOS women. In this randomised controlled clinical trial, 60 women with PCOS were randomly assigned to one of two diets with energy restriction: the DASH diet and a control diet. The DASH and control diets consisted of 50-55% carbohydrate, 15-20% protein and 25-30% total fat. The DASH diet was designed to be rich in vegetables, fruits, whole grains and low-fat dairy products, as well as low in saturated fats, cholesterol, refined grains and sweets. In the present study, the anthropometric indices, body composition, total testosterone, androstenedione, sex hormone binding globulin (SHBG), free androgen index and 2,2'-diphenyl-1-picryylhydrazyl (DPPH) scavenging activity were measured before and after 3 months. The consumption of DASH diet compared to the control diet was associated with a significant reduction in weight [-5.78 (1.91) kg versus -4.34 (2.87) kg, P = 0.032], body mass index (BMI) [-2.29 (0.15) kg m -2 versus -1.69 (0.20) kg m -2 , P = 0.02], fat mass [-3.23(1.66) kg versus -2.13 (1.26) kg, P = 0.008] and serum androstenedione [-1.75 (1.39) ng mL -1 versus -1.02 (0.72) ng mL -1 , P-value = 0.019]. Increased concentrations of SHBG [28.80 (21.71) versus 11.66(18.82) nmol L -1 , P = 0.003) and DPPH scavenging activity [30.23% (19.09) versus 12.97% (25.12) were also found in the DASH group. The DASH diet could improve weight loss, BMI and fat mass. Furthermore, it could result in a significant reduction in serum androstenedione and a significant increase in antioxidant status and SHBG. © 2016 The British Dietetic Association Ltd.

Dietary habits, poverty, and chronic kidney disease in an urban population.

PubMed

Crews, Deidra C; Kuczmarski, Marie Fanelli; Miller, Edgar R; Zonderman, Alan B; Evans, Michele K; Powe, Neil R

2015-03-01

Poverty is associated with chronic kidney disease (CKD) in the United States and worldwide. Poor dietary habits may contribute to this disparity. Cross-sectional study. A total of 2,058 community-dwelling adults aged 30 to 64 years residing in Baltimore City, Maryland. Adherence to the Dietary Approaches to Stop Hypertension (DASH) diet. DASH scoring based on 9 target nutrients (total fat, saturated fat, protein, fiber, cholesterol, calcium, magnesium, sodium, and potassium); adherence defined as score ≥4.5 of maximum possible score of 9. Poverty (self-reported household income <125% of 2004 Department of Health and Human Services guideline) and nonpoverty (≥125% of guideline). CKD defined as estimated glomerular filtration rate <60 mL/minute/1.73 m(2) (CKD epidemiology collaboration equation). Multivariable logistic regression was used to calculate adjusted odds ratios (AORs) for relation of DASH score tertile and CKD, stratified by poverty status. Among 2,058 participants (mean age 48 years; 57% black; 44% male; 42% with poverty), median DASH score was low, 1.5 (interquartile range, 1-2.5). Only 5.4% were adherent. Poverty, male sex, black race, and smoking were more prevalent among the lower DASH score tertiles, whereas higher education and regular health care were more prevalent among the highest DASH score tertile (P < .05 for all). Fiber, calcium, magnesium, and potassium intake were lower, and cholesterol higher, among the poverty compared with nonpoverty group (P < .05 for all), with no difference in sodium intake. A total of 5.6% of the poverty and 3.8% of the nonpoverty group had CKD (P = .05). The lowest DASH tertile (compared with the highest) was associated with more CKD among the poverty (AOR 3.15, 95% confidence interval 1.51-6.56), but not among the nonpoverty group (AOR 0.73, 95% confidence interval 0.37-1.43; P interaction = .001). Poor dietary habits are strongly associated with CKD among the urban poor and may represent a target for interventions aimed at reducing disparities in CKD. Copyright © 2015 National Kidney Foundation, Inc. All rights reserved.

Dietary Habits, Poverty, and Chronic Kidney Disease in an Urban Population

PubMed Central

Crews, Deidra C.; Kuczmarski, Marie Fanelli; III, Edgar R. Miller; Zonderman, Alan B.; Evans, Michele K.; Powe, Neil R.

2014-01-01

Background Poverty is associated with chronic kidney disease (CKD) in the US and worldwide. Poor dietary habits may contribute to this disparity. Study Design Cross-sectional study. Setting & Participants 2,058 community-dwelling adults aged 30-64 years residing in Baltimore City, Maryland. Predictors Adherence to the Dietary Approaches to Stop Hypertension (DASH) diet. DASH scoring based on 9 target nutrients (total fat, saturated fat, protein, fiber, cholesterol, calcium, magnesium, sodium, and potassium); adherence defined as score ≥4.5 out of maximum possible score of 9. Poverty (self-reported household income <125% of 2004 Department of Health and Human Services guideline) and non-poverty (≥125% of guideline). Outcomes & Measurements CKD defined as estimated glomerular filtration rate <60mL/min/1.73m2 (CKD-EPI). Multivariable logistic regression used to calculate adjusted odds ratios (AORs) for relation of DASH score tertile and CKD, stratified by poverty status. Results Among 2,058 participants (mean age 48 years; 57% black; 44% male; 42% with poverty), median DASH score was low, 1.5 (IQR, 1-2.5). Only 5.4% were adherent. Poverty, male sex, black race, and smoking were more prevalent among the lower DASH score tertiles, while higher education and regular health care were more prevalent among the highest DASH score tertile (P<0.05 for all). Fiber, calcium, magnesium and potassium intake were lower, and cholesterol higher, among the poverty as compared to non-poverty group (P<0.05 for all), with no difference in sodium intake. A total of 5.6% of the poverty and 3.8% of the non-poverty group had CKD (P=0.05). The lowest DASH tertile (compared to the highest) was associated with more CKD among the poverty [AOR 3.15, 95% Confidence Interval (CI) 1.51-6.56], but not among the non-poverty group (AOR 0.73, 95% CI 0.37-1.43). P interaction 0.001. Conclusions Poor dietary habits are strongly associated with CKD among the urban poor and may represent a target for interventions aimed at reducing disparities in CKD. PMID:25238697

Identification and Partial Characterization of Extracellular Aspartic Protease Genes from Metschnikowia pulcherrima IWBT Y1123 and Candida apicola IWBT Y1384

PubMed Central

Reid, Vernita J.; Theron, Louwrens W.; du Toit, Maret

2012-01-01

The extracellular acid proteases of non-Saccharomyces wine yeasts may fulfill a number of roles in winemaking, which include increasing the available nitrogen sources for the growth of fermentative microbes, affecting the aroma profile of the wine, and potentially reducing protein haze formation. These proteases, however, remain poorly characterized, especially at genetic level. In this study, two extracellular aspartic protease-encoding genes were identified and sequenced, from two yeast species of enological origin: one gene from Metschnikowia pulcherrima IWBT Y1123, named MpAPr1, and the other gene from Candida apicola IWBT Y1384, named CaAPr1. In silico analysis of these two genes revealed a number of features peculiar to aspartic protease genes, and both the MpAPr1 and CaAPr1 putative proteins showed homology to proteases of yeast genera. Heterologous expression of MpAPr1 in Saccharomyces cerevisiae YHUM272 confirmed that it encodes an aspartic protease. MpAPr1 production, which was shown to be constitutive, and secretion were confirmed in the presence of bovine serum albumin (BSA), casein, and grape juice proteins. The MpAPr1 gene was found to be present in 12 other M. pulcherrima strains; however, plate assays revealed that the intensity of protease activity was strain dependent and unrelated to the gene sequence. PMID:22820332

Partial purification of histone H3 proteolytic activity from the budding yeast Saccharomyces cerevisiae.

PubMed

Azad, Gajendra Kumar; Tomar, Raghuvir Singh

2016-06-01

The proteolytic clipping of histone tails has recently emerged as a novel form of irreversible post-translational modification (PTM) of histones. Histone clipping has been implicated as a regulatory process leading to the permanent removal of PTMs from histone proteins. However, there is scarcity of literature that describes the identification and characterization of histone-specific proteases. Here, we employed various biochemical methods to report histone H3-specific proteolytic activity from budding yeast. Our results demonstrate that H3 proteolytic activity was associated with sepharose bead matrices and activity was not affected by a variety of stress conditions. We have also identified the existence of an unknown protein that acts as a physiological inhibitor of the H3-clipping activity of yeast H3 protease. Moreover, through protease inhibition assays, we have also characterized yeast H3 protease as a serine protease. Interestingly, unlike glutamate dehydrogenase (GDH), yeast H3 proteolytic activity was not inhibited by Stefin B. Together, our findings suggest the existence of a novel H3 protease in yeast that is different from other reported histone H3 proteases. The presence of histone H3 proteolytic activity, along with the physiological inhibitor in yeast, suggests an interesting molecular mechanism that regulates the activity of histone proteases. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Effects of Different Conditioning Activities on 100-m Dash Performance in High School Track and Field Athletes.

PubMed

Ferreira-Júnior, João B; Guttierres, Ana P M; Encarnação, Irismar G A; Lima, Jorge R P; Borba, Diego A; Freitas, Eduardo D S; Bemben, Michael G; Vieira, Carlos A; Bottaro, Martim

2018-06-01

This study compared the effects of different conditioning activities on the 100-m dash performance of 11 male, high school track and field athletes (mean age = 16.3; SD = 1.2 years). Participants performed a 100-m dash seven minutes after each of four randomized conditioning protocols, with each condition and 100-m dash separated by 3-10 days. The conditioning protocols were (a) control, no conditioning activity; (b) weighted plyometric, three sets of 10 repetitions of alternate leg bounding with additional load of 10% of the body mass; (c) free sprint, two 20-m sprints; and (d) resisted sprint (RS), two 20-m resisted sprints using an elastic tubing tool. We obtained session ratings of perceived exertion (SRPE) immediately after each conditioning protocol. There were no significant differences between any of the three experimental conditioning activities on 100-m sprint time, but the RS protocol improved 100-m sprint time compared with the control (no conditioning) protocol ( p < .001). The RS also led to greater sprint velocity and higher SRPE compared with the control condition ( p < .01). There was no significant association between SRPE and 100-m performance ( p = .77, r = .05). These results suggest a benefit for young male track and field athletes to the elastic tubing warm-up activities prior to the 100-m dash.

Purification, Characterization, and Functional Role of a Novel Extracellular Protease from Pleurotus ostreatus

PubMed Central

Palmieri, Gianna; Bianco, Carmen; Cennamo, Giovanna; Giardina, Paola; Marino, Gennaro; Monti, Maria; Sannia, Giovanni

2001-01-01

A new extracellular protease (PoSl; Pleurotus ostreatus subtilisin-like protease) from P. ostreatus culture broth has been purified and characterized. PoSl is a monomeric glycoprotein with a molecular mass of 75 kDa, a pI of 4.5, and an optimum pH in the alkaline range. The inhibitory profile indicates that PoSl is a serine protease. The N-terminal and three tryptic peptide sequences of PoSl have been determined. The homology of one internal peptide with conserved sequence around the Asp residue of the catalytic triad in the subtilase family suggests that PoSl is a subtilisin-like protease. This hypothesis is further supported by the finding that PoSl hydrolysis sites of the insulin B chain match those of subtilisin. PoSl activity is positively affected by calcium. A 10-fold decrease in the Km value in the presence of calcium ions can reflect an induced structural change in the substrate recognition site region. Furthermore, Ca2+ binding slows PoSl autolysis, triggering the protein to form a more compact structure. These effects have already been observed for subtilisin and other serine proteases. Moreover, PoSl protease seems to play a key role in the regulation of P. ostreatus laccase activity by degrading and/or activating different isoenzymes. PMID:11375191

Protein quality control in organelles - AAA/FtsH story.

PubMed

Janska, Hanna; Kwasniak, Malgorzata; Szczepanowska, Joanna

2013-02-01

This review focuses on organellar AAA/FtsH proteases, whose proteolytic and chaperone-like activity is a crucial component of the protein quality control systems of mitochondrial and chloroplast membranes. We compare the AAA/FtsH proteases from yeast, mammals and plants. The nature of the complexes formed by AAA/FtsH proteases and the current view on their involvement in degradation of non-native organellar proteins or assembly of membrane complexes are discussed. Additional functions of AAA proteases not directly connected with protein quality control found in yeast and mammals but not yet in plants are also described shortly. Following an overview of the molecular functions of the AAA/FtsH proteases we discuss physiological consequences of their inactivation in yeast, mammals and plants. The molecular basis of phenotypes associated with inactivation of the AAA/FtsH proteases is not fully understood yet, with the notable exception of those observed in m-AAA protease-deficient yeast cells, which are caused by impaired maturation of mitochondrial ribosomal protein. Finally, examples of cytosolic events affecting protein quality control in mitochondria and chloroplasts are given. This article is part of a Special Issue entitled: Protein Import and Quality Control in Mitochondria and Plastids. Copyright © 2012 Elsevier B.V. All rights reserved.

Peptidomimetic Escape Mechanisms Arise via Genetic Diversity in the Ligand-Binding Site of the Hepatitis C Virus NS3/4A Serine Protease

PubMed Central

Welsch, Christoph; Shimakami, Tetsuro; Hartmann, Christoph; Yang, Yan; Domingues, Francisco S.; Lengauer, Thomas; Zeuzem, Stefan; Lemon, Stanley M.

2011-01-01

Background & Aims It is a challenge to develop direct-acting antiviral agents (DAAs) that target the NS3/4A protease of hepatitis C virus (HCV) because resistant variants develop. Ketoamide compounds, designed to mimic the natural protease substrate, have been developed as inhibitors. However, clinical trials have revealed rapid selection of resistant mutants, most of which are considered to be pre-existing variants. Methods We identified residues near the ketoamide-binding site in X-ray structures of the genotype 1a protease, co-crystallized with boceprevir or a telaprevir-like ligand, and then identified variants at these positions in 219 genotype 1 sequences from a public database. We used side-chain modeling to assess the potential effects of these variants on the interaction between ketoamide and the protease, and compared these results with the phenotypic effects on ketoamide resistance, RNA replication capacity, and infectious virus yields in a cell culture model of infection. Results Thirteen natural binding-site variants with potential for ketoamide resistance were identified at 10 residues in the protease, near the ketoamide binding site. Rotamer analysis of amino acid side-chain conformations indicated that 2 variants (R155K and D168G) could affect binding of telaprevir more than boceprevir. Measurements of antiviral susceptibility in cell culture studies were consistent with this observation. Four variants (Q41H, I132V, R155K, and D168G) caused low-to-moderate levels of ketoamide resistance; 3 of these were highly fit (Q41H, I132V, and R155K). Conclusions Using a comprehensive sequence and structure-based analysis, we showed how natural variation in the HCV protease NS3/4A sequences might affect susceptibility to first-generation DAAs. These findings increase our understanding of the molecular basis of ketoamide resistance among naturally existing viral variants. PMID:22155364

Modification of host erythrocyte membranes by trypsin and chymotrypsin treatments and effects on the in vitro growth of bovine and equine Babesia parasites.

PubMed

Okamura, Masashi; Yokoyama, Naoaki; Takabatake, Noriyuki; Okubo, Kazuhiro; Ikehara, Yuzuru; Igarashi, Ikuo

2007-02-01

In the present study, we investigated the effects of protease pretreatments of host erythrocytes (RBC) on the in vitro growth of bovine Babesia parasites (Babesia bovis and B. bigemina) and equine Babesia parasites (B. equi and B. caballi). The selected proteases, trypsin and chymotrypsin, clearly modified several membrane proteins of both bovine and equine RBC, as demonstrated by SDS-PAGE analysis; however, the protease treatments also modified the sialic acid content exclusively in bovine RBC, as demonstrated by lectin blot analysis. An in vitro growth assay using the protease-treated RBC showed that the trypsin-treated bovine RBC, but not the chymotrypsin-treated ones, significantly reduced the growth of B. bovis and B. bigemina as compared to the control. In contrast, the growth of B. equi and B. caballi was not affected by any of these proteases. Thus, the bovine, but not the equine, Babesia parasites require the trypsin-sensitive membrane (sialoglyco) proteins to infect the RBC.

Generic protease detection technology for monitoring periodontal disease.

PubMed

Zheng, Xinwei; Cook, Joseph P; Watkinson, Michael; Yang, Shoufeng; Douglas, Ian; Rawlinson, Andrew; Krause, Steffi

2011-01-01

Periodontal diseases are inflammatory conditions that affect the supporting tissues of teeth and can lead to destruction of the bone support and ultimately tooth loss if untreated. Progression of periodontitis is usually site specific but not uniform, and currently there are no accurate clinical methods for distinguishing sites where there is active disease progression from sites that are quiescent. Consequently, unnecessary and costly treatment of periodontal sites that are not progressing may occur. Three proteases have been identified as suitable markers for distinguishing sites with active disease progression and quiescent sites: human neutrophil elastase, cathepsin G and MMP8. Generic sensor materials for the detection of these three proteases have been developed based on thin dextran hydrogel films cross-linked with peptides. Degradation of the hydrogel films was monitored using impedance measurements. The target proteases were detected in the clinically relevant range within a time frame of 3 min. Good specificity for different proteases was achieved by choosing appropriate peptide cross-linkers.

78 FR 21573 - Airworthiness Directives; Bombardier, Inc. Airplanes

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-11

.... For service information identified in this proposed AD, contact Bombardier, Inc., Q-Series Technical... following service information: de Havilland Dash 8 Series 100 Maintenance Task Card 531X1, Revision 25, in... Havilland Dash 8 Series 100 Maintenance Task Card 631X1, Revision 25, in Section 8, Electrical Wiring...

75 FR 45496 - Amendment of the Commission's Rules to Accommodate 30 Megahertz Channels in the 6525-6875 MHz...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-03

... to Provide for Conditional Authorization on Additional Channels in the 21.8[dash]22.0 GHz and 23.0[dash]23.2 GHz Band Correction In rule document 2010-17205 beginning on page 41767 in the issue of...

Microbial degradation at a shallow coastal site: Long-term spectra and rates of exoenzymatic activities in the NE Adriatic Sea

NASA Astrophysics Data System (ADS)

Celussi, Mauro; Del Negro, Paola

2012-12-01

The degradation of organic matter along the water column is mediated by enzymes released into the environment by planktonic organisms. Variations in enzymes profiles (types and levels of activity) reflect the trophic status of the environment and could be caused by shifts in the dominant species or in the level of enzyme expression by the same species in response to changes in the spectrum of organic substrates. To explore this issue, we examined the maximum rates of hydrolysis of 6 different enzymes (protease, α-glucosidase, β-glucosidase, β-galactosidase, alkaline phosphatase and lipase) along the water column (4 depths) at a coastal station in the Gulf of Trieste (northern Adriatic Sea), from 2000 to 2005. Most of the studied enzymes exhibited a pronounced seasonal variability with winter minima and maxima from April to October. During summer, alkaline phosphatase, lipase and protease reached the highest activities, while polysaccharide degradation prevailed in spring and autumn, associated to phytoplankton blooms. Phosphatase/protease activities ratio was generally low, indicating that microbial communities were rarely P-limited, possibly because of the use of organic P sources. A pronounced interannual variability of degradation patterns was found, with maximum rates of protease being the highest in most of the samples, followed by the alkaline phosphatase's ones. Water column features greatly affected hydrolysis rates, being degradation of linear polysaccharides, lipids, phosphorilated compounds and polypeptides significantly different at different depths during stratified condition. Mixing processes affected especially α-glucosidase activity, possibly as a consequence of resuspension of organic matter from the seabed. Large-impact phenomena such as the 2003 heat wave and mucilage influenced the degradation of specific substrates. Mucilage enhanced lipase, phosphatase and protease, whereas a pronounced inhibition characterised phosphatase and protease during summer 2003.

Detergent-compatible, organic solvent-tolerant alkaline protease from Bacillus circulans MTCC 7942: Purification and characterization.

PubMed

Patil, Ulhas; Mokashe, Narendra; Chaudhari, Ambalal

2016-01-01

Proteases are now recognized as the most indispensable industrial biocatalyst owing to their diverse microbial sources and innovative applications. In the present investigation, a thermostable, organic solvent-tolerant, alkaline serine protease from Bacillus circulans MTCC 7942, was purified and characterized. The protease was purified to 37-fold by a three-step purification scheme with 39% recovery. The optimum pH and temperature for protease was 10 and 60 °C, respectively. The apparent molecular mass of the purified enzyme was 43 kD as revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The Km and Vmax values using casein-substrate were 3.1 mg/mL and 1.8 µmol/min, respectively. The protease remained stable in the presence of organic solvents with higher (>3.2) log P value (cyclohexane, n-octane, n-hexadecane, n-decane, and n-dodecane), as compared to organic solvents with lower (<3.2) log P value (acetone, butanol, benzene, chloroform, toluene). Remarkably, the protease showed profound stability even in the presence of organic solvents with less log P values (glycerol, dimethyl sulfate [DMSO], p-xylene), indicating the possibility of nonaqueous enzymatic applications. Also, protease activity was improved in the presence of metal ions (Ca(2+), Mg(2+), Mn(2+)); enhanced by biosurfactants; hardly affected by bleaching agents, oxidizing agents, and chemical surfactants; and stable in commercial detergents. In addition, a protease-detergent formulation effectively washed out egg and blood stains as compared to detergent alone. The protease was suitable for various commercial applications like processing of gelatinous film and as a compatible additive to detergent formulation with its operative utility in hard water.

Radiological and clinical predictors of long-term outcome in rotator cuff calcific tendinitis.

PubMed

de Witte, Pieter Bas; van Adrichem, Raymond A; Selten, Jasmijn W; Nagels, Jochem; Reijnierse, M; Nelissen, Rob G H H

2016-10-01

Knowledge on the epidemiology and long-term course of rotator cuff calcific tendinitis (RCCT) is scarce. We assessed demographics, radiological characteristics, and their association with long-term outcomes in a large patient group. Baseline demographics, radiological characteristics and treatment were recorded in 342 patients. Interobserver agreement of radiological measures was analyzed. Long-term outcome was evaluated with questionnaires (WORC, DASH). The association of baseline characteristics with outcome was assessed. Mean age was 49.0 (SD = 10.0), and 59.5 % were female. The dominant arm was affected in 66.0 %, and 21.3 % had bilateral disease. Calcifications were on average 18.7 mm (SD = 10.1, ICC = 0.84 (p < 0.001)) and located 10.1 mm (SD = 11.8) medially to the acromion (ICC = 0.77 (p < 0.001)). Gärtner type I calcifications were found in 32.1 % (Kappa = 0.47 (p < 0.001)). After 14 years (SD = 7.1) of follow-up, median WORC was 72.5 (range, 3.0-100.0; WORC < 60 in 42 %) and median DASH 17.0 (range, 0.0-82.0). Female gender, dominant arm involvement, bilateral disease, longer duration of symptoms, and multiple calcifications were associated with inferior WORC. DASH results were similar. Many subjects have persisting shoulder complaints years after diagnosis, regardless of treatment. Female gender, dominant arm involvement, bilateral disease, longer duration of symptoms, and multiple calcifications were associated with inferior outcome. Radiological measures had moderate-to-good reliability and no prognostic value. • Most RCCT studies report on short-term outcome and/or small patients groups. • In this large, long-term observational study, RCCT appeared to not be self-limiting in many subjects. • Negative prognostic factors included female gender, more calcifications, dominant arm affected, and longer duration of symptoms. • Interobserver agreement of general radiological RCCT measures is moderate to good. • More rigorous diagnostics and treatment might be needed in specific RCCT cases.

The serine protease inhibitor TLCK attenuates intrinsic death pathways in neurons upstream of mitochondrial demise.

PubMed

Reuther, C; Ganjam, G K; Dolga, A M; Culmsee, C

2014-11-01

It is well-established that activation of proteases, such as caspases, calpains and cathepsins are essential components in signaling pathways of programmed cell death (PCD). Although these proteases have also been linked to mechanisms of neuronal cell death, they are dispensable in paradigms of intrinsic death pathways, e.g. induced by oxidative stress. However, emerging evidence implicated a particular role for serine proteases in mechanisms of PCD in neurons. Here, we investigated the role of trypsin-like serine proteases in a model of glutamate toxicity in HT-22 cells. In these cells glutamate induces oxytosis, a form of caspase-independent cell death that involves activation of the pro-apoptotic protein BH3 interacting-domain death agonist (Bid), leading to mitochondrial demise and ensuing cell death. In this model system, the trypsin-like serine protease inhibitor Nα-tosyl-l-lysine chloromethyl ketone hydrochloride (TLCK) inhibited mitochondrial damage and cell death. Mitochondrial morphology alterations, the impairment of the mitochondrial membrane potential and ATP depletion were prevented and, moreover, lipid peroxidation induced by glutamate was completely abolished. Strikingly, truncated Bid-induced cell death was not affected by TLCK, suggesting a detrimental activity of serine proteases upstream of Bid activation and mitochondrial demise. In summary, this study demonstrates the protective effect of serine protease inhibition by TLCK against oxytosis-induced mitochondrial damage and cell death. These findings indicate that TLCK-sensitive serine proteases play a crucial role in cell death mechanisms upstream of mitochondrial demise and thus, may serve as therapeutic targets in diseases, where oxidative stress and intrinsic pathways of PCD mediate neuronal cell death.

Nanostructured Quantum Dots or Dashes in Photovoltaic Devices and Methods Thereof

NASA Technical Reports Server (NTRS)

Raffaele, Ryne P. (Inventor); Wilt, David M. (Inventor)

2015-01-01

A photovoltaic device includes one or more structures, an array of at least one of quantum dots and quantum dashes, at least one groove, and at least one conductor. Each of the structures comprises an intrinsic layer on one of an n type layer and a p type layer and the other one of the n type layer and the p type layer on the intrinsic layer. The array of at least one of quantum dots and quantum dashes is located in the intrinsic layer in at least one of the structures. The groove extends into at least one of the structures and the conductor is located along at least a portion of the groove.

Expression of Bacillus protease (Protease BYA) from Bacillus sp. Y in Bacillus subtilis and enhancement of its specific activity by site-directed mutagenesis-improvement in productivity of detergent enzyme-.

PubMed

Tobe, Seiichi; Shimogaki, Hisao; Ohdera, Motoyasu; Asai, Yoshio; Oba, Kenkichi; Iwama, Masanori; Irie, Masachika

2006-01-01

An attempt was made to express protease BYA produced by an alkalophilic Bacillus sp. Y in Bacillus subtilis by gene engineering methods. The gene encoding protease BYA was cloned from Bacillus sp. Y, and expression vector pTA71 was constructed from the amylase promoter of Bacillus licheniformis, DNA fragments encoding the open reading frame of protease BYA, and pUB110. Protease BYA was secreted at an activity level of 5100 APU/ml in the common industrial culture medium of Bacillus subtilis transformed with pTA71. We then attempted to increase the specific activity of protease BYA by site-directed mutagenesis. Amino acid residue Ala29 next to catalytic Asp30 was replaced by one of three uncharged amino acid residues (Val29, Leu29, Ile29), and each mutant enzyme was expressed and isolated from the culture medium. Val29 mutant enzyme was secreted at an activity level of greater than 7000 APU/ml in culture medium, and its specific activity was 1.5-fold higher than that of the wild-type enzyme. Other mutant enzymes had specific activity similar to that of the original one and were less stabile than the wild-type enzyme. It can be thought that the substitution at amino acid residue 29 affects the level of activity and stability of protease BYA.

77 FR 63215 - Airworthiness Directives; Bombardier, Inc. Airplanes

Federal Register 2010, 2011, 2012, 2013, 2014

2012-10-16

... can be found in the Bombardier Q400 Dash 8 Aircraft Maintenance Manual. (h) Retained Actions for... operational check fails, guidance on doing the repair can be found in the Bombardier Q400 Dash 8 Aircraft... which the reidentification is done, an operational check of the alternate extension system of the main...

77 FR 67763 - Airworthiness Directives; Pratt & Whitney Division Turbofan Engines

Federal Register 2010, 2011, 2012, 2013, 2014

2012-11-14

... Airworthiness Directives; Pratt & Whitney Division Turbofan Engines AGENCY: Federal Aviation Administration (FAA..., PW4156A, PW4158, PW4160, PW4460, PW4462, and PW4650 turbofan engines, including models with any dash... PW4650 turbofan engines, including models with any dash number suffix, with 3rd stage low-pressure...

Validity of the Sleep Subscale of the Diagnostic Assessment for the Severely Handicapped-II (DASH-II)

ERIC Educational Resources Information Center

Matson, Johnny L.; Malone, Carrie J.

2006-01-01

Currently there are no available sleep disorder measures for individuals with severe and profound intellectual disability. We, therefore, attempted to establish the external validity of the "Diagnostic Assessment for the Severely Handicapped-II" (DASH-II) sleep subscale by comparing daily observational sleep data with the responses of…

Hermaphroditism in the dash-and-dot goatfish Parupeneus barberinus.

PubMed

Longenecker, K; Langston, R; Crane, M

2017-03-01

Presence of bisexual individuals and a sex-specific bimodal size distribution are suggestive of protogyny in the dash-and-dot goatfish Parupeneus barberinus, but the most parsimonious interpretation of histological analysis is juvenile hermaphroditism. This is the first report of hermaphroditism in the Mullidae. © 2017 The Fisheries Society of the British Isles.

Development of DASH Mobile: a mHealth lifestyle change intervention for the management of hypertension.

PubMed

Mann, Devin M; Kudesia, Valmeek; Reddy, Shivani; Weng, Michael; Imler, Daniel; Quintiliani, Lisa

2013-01-01

Several landmark studies based on the DASH diet have established the effectiveness of a lifestyle approach to blood pressure control that emphasizes a diet rich in fruits and vegetables with moderate portions of low-fat dairy and lean protein along with increased physical activity and reduced sodium intake. However, this evidence base remains underused due feasibility limitations of implementing these intense in-person interventions and poor engagement with desktop computer based versions. Mobile technologies such as smartphones and wireless sensors have the ability to deliver behavioral interventions in-the-moment and with reduced user burden. DASH Mobile is a new mHealth system being developed to deliver this evidence-based lifestyle intervention to hypertensive patients. The system consists of an Android based "app" that facilitates easy tracking of DASH food portions, integrated Bluetooth blood pressure, weight and pedometer monitoring, goal setting, simple data visualizations and multimedia video clips to train patients in the basic concepts of the lifestyle change plan. At present, the system is undergoing usability testing with a pilot clinical trial planned for Spring 2013.

Hydridosilylamido complexes of Ta and Mo isolobal with Berry's zirconocenes: syntheses, β-Si-H agostic interactions, catalytic hydrosilylation, and insight into mechanism.

PubMed

McLeod, Nicolas A; Kuzmina, Lyudmila G; Korobkov, Ilia; Howard, Judith A K; Nikonov, Georgii I

2016-02-14

The syntheses of novel Group 5 and Group 6 hydrosilylamido complexes of the type R(ArN[double bond, length as m-dash])M{N((t)Bu)SiMe2-H}X (M = Ta, R = Cp; M = Mo, R = ArN; X = Cl, H, OBn, Me) are described. The various substituents in the X position seem to play the key role in determining the extent of β-agostic interaction with the Si-H bond. The Mo agostic hydrido complex (ArN[double bond, length as m-dash])2Mo{η(3)-N((t)Bu)SiMe2-H}H is a pre-catalyst for the hydrosilylation of carbonyls. The stoichiometric reaction between benzaldehyde and (ArN[double bond, length as m-dash])2Mo{η(3)-N((t)Bu)SiMe2-H}H gives the benzoxy complex (ArN[double bond, length as m-dash])2Mo{N((t)Bu)SiMe2-H}(OBn), which showed a similar catalytic reactivity compared to the parent hydride. Mechanistic studies suggest that a non-hydride mechanism is operative.

Highly efficient non-degenerate four-wave mixing under dual-mode injection in InP/InAs quantum-dash and quantum-dot lasers at 1.55 μm

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sadeev, T., E-mail: tagir@mailbox.tu-berlin.de; Arsenijević, D.; Huang, H.

2015-11-09

This work reports on non-degenerate four-wave mixing under dual-mode injection in metalorganic vapor phase epitaxy grown InP/InAs quantum-dash and quantum dot Fabry-Perot laser operating at 1550 nm. High values of normalized conversion efficiency of −18.6 dB, optical signal-to-noise ratio of 37 dB, and third order optical susceptibility normalized to material gain χ{sup (3)}/g{sub 0} of ∼4 × 10{sup −19} m{sup 3}/V{sup 3} are measured for 1490 μm long quantum-dash lasers. These values are similar to those obtained with distributed-feedback lasers and semiconductor optical amplifiers, which are much more complicated to fabricate. On the other hand, due to the faster gain saturation and enhanced modulation of carriermore » populations, quantum-dot lasers demonstrate 12 dB lower conversion efficiency and 4 times lower χ{sup (3)}/g{sub 0} compared to quantum dash lasers.« less

Data Access System for Hydrology

NASA Astrophysics Data System (ADS)

Whitenack, T.; Zaslavsky, I.; Valentine, D.; Djokic, D.

2007-12-01

As part of the CUAHSI HIS (Consortium of Universities for the Advancement of Hydrologic Science, Inc., Hydrologic Information System), the CUAHSI HIS team has developed Data Access System for Hydrology or DASH. DASH is based on commercial off the shelf technology, which has been developed in conjunction with a commercial partner, ESRI. DASH is a web-based user interface, developed in ASP.NET developed using ESRI ArcGIS Server 9.2 that represents a mapping, querying and data retrieval interface over observation and GIS databases, and web services. This is the front end application for the CUAHSI Hydrologic Information System Server. The HIS Server is a software stack that organizes observation databases, geographic data layers, data importing and management tools, and online user interfaces such as the DASH application, into a flexible multi- tier application for serving both national-level and locally-maintained observation data. The user interface of the DASH web application allows online users to query observation networks by location and attributes, selecting stations in a user-specified area where a particular variable was measured during a given time interval. Once one or more stations and variables are selected, the user can retrieve and download the observation data for further off-line analysis. The DASH application is highly configurable. The mapping interface can be configured to display map services from multiple sources in multiple formats, including ArcGIS Server, ArcIMS, and WMS. The observation network data is configured in an XML file where you specify the network's web service location and its corresponding map layer. Upon initial deployment, two national level observation networks (USGS NWIS daily values and USGS NWIS Instantaneous values) are already pre-configured. There is also an optional login page which can be used to restrict access as well as providing a alternative to immediate downloads. For large request, users would be notified via email with a link to their data when it is ready.

Unravelling the immunological roles of dipeptidyl peptidase 4 (DPP4) activity and/or structure homologue (DASH) proteins

PubMed Central

Klemann, C.; Stephan, M.

2016-01-01

Summary Dipeptidyl peptidase (DPP) 4 (CD26, DPP4) is a multi‐functional protein involved in T cell activation by co‐stimulation via its association with adenosine deaminase (ADA), caveolin‐1, CARMA‐1, CD45, mannose‐6‐phosphate/insulin growth factor‐II receptor (M6P/IGFII‐R) and C‐X‐C motif receptor 4 (CXC‐R4). The proline‐specific dipeptidyl peptidase also modulates the bioactivity of several chemokines. However, a number of enzymes displaying either DPP4‐like activities or representing structural homologues have been discovered in the past two decades and are referred to as DPP4 activity and/or structure homologue (DASH) proteins. Apart from DPP4, DASH proteins include fibroblast activation protein alpha (FAP), DPP8, DPP9, DPP4‐like protein 1 (DPL1, DPP6, DPPX L, DPPX S), DPP4‐like protein 2 (DPL2, DPP10) from the DPP4‐gene family S9b and structurally unrelated enzyme DPP2, displaying DPP4‐like activity. In contrast, DPP6 and DPP10 lack enzymatic DPP4‐like activity. These DASH proteins play important roles in the immune system involving quiescence (DPP2), proliferation (DPP8/DPP9), antigen‐presenting (DPP9), co‐stimulation (DPP4), T cell activation (DPP4), signal transduction (DPP4, DPP8 and DPP9), differentiation (DPP4, DPP8) and tissue remodelling (DPP4, FAP). Thus, they are involved in many pathophysiological processes and have therefore been proposed for potential biomarkers or even drug targets in various cancers (DPP4 and FAP) and inflammatory diseases (DPP4, DPP8/DPP9). However, they also pose the challenge of drug selectivity concerning other DASH members for better efficacy and/or avoidance of unwanted side effects. Therefore, this review unravels the complex roles of DASH proteins in immunology. PMID:26671446

The effects of dietary approaches to stop hypertension diet on weight loss, anti-Müllerian hormone and metabolic profiles in women with polycystic ovary syndrome: A randomized clinical trial.

PubMed

Foroozanfard, Fatemeh; Rafiei, Hamideh; Samimi, Mansooreh; Gilasi, Hamid Reza; Gorjizadeh, Roohangiz; Heidar, Zahra; Asemi, Zatollah

2017-07-01

This study was designed to evaluate the effects of the dietary approaches to stop hypertension (DASH diet) on weight loss, anti-Müllerian hormone (AMH) and metabolic profiles in women with polycystic ovary syndrome (PCOS). A randomized controlled clinical trial was conducted among 60 overweight or obese patients with PCOS. Patients were randomly assigned to receive either low-calorie DASH (N=30) or control diet (N=30) for 12 weeks. The DASH and control diets were consisted of 52%-55% carbohydrates, 16%-18% proteins and 30% total fats; however, the DASH diet was designed to be rich in fruits, vegetables, whole grains, low-fat dairy products, cholesterol and refined grains. Both diets were equicaloric. Adherence to the DASH diet, compared to the control diet, resulted in a significant decrease in BMI (-1.6±0.5 vs -1.2±0.7 kg/m 2 , P=.02). Significant decreases in AMH (-1.1±3.1 vs +0.3±0.7 ng/mL, P=.01), insulin (-25.2±51.0 vs -1.2±28.8 pmol/L, P=.02), homoeostasis model of assessment-estimated insulin resistance (-0.9±2.0 vs -0.1±1.0, P=.02), free androgen index (FAI; -0.03±0.09 vs +0.06±0.21, P=.02) and malondialdehyde (MDA) levels (-0.5±0.4 vs +0.2±0.3 μmol/L, P<.001), and significant increases in quantitative insulin sensitivity check index (+0.01±0.03 vs -0.004±0.01, P=.02), sex hormone-binding globulin (SHBG; +3.7±8.5 vs -1.5±7.2 nmol/L, P=.01) and nitric oxide (NO; +9.0±4.9 vs +0.6±2.3 μmol/L, P<.001) were also seen in the DASH group compared with the control group. Adherence to the DASH diet for 12 weeks among PCOS women had beneficial effects on BMI, AMH, insulin metabolism, SHBG, FAI, NO and MDA levels. © 2017 John Wiley & Sons Ltd.

Multiple Off-Ice Performance Variables Predict On-Ice Skating Performance in Male and Female Division III Ice Hockey Players.

PubMed

Janot, Jeffrey M; Beltz, Nicholas M; Dalleck, Lance D

2015-09-01

The purpose of this study was to determine if off-ice performance variables could predict on-ice skating performance in Division III collegiate hockey players. Both men (n = 15) and women (n = 11) hockey players (age = 20.5 ± 1.4 years) participated in the study. The skating tests were agility cornering S-turn, 6.10 m acceleration, 44.80 m speed, modified repeat skate, and 15.20 m full speed. Off-ice variables assessed were years of playing experience, height, weight and percent body fat and off-ice performance variables included vertical jump (VJ), 40-yd dash (36.58m), 1-RM squat, pro-agility, Wingate peak power and peak power percentage drop (% drop), and 1.5 mile (2.4km) run. Results indicated that 40-yd dash (36.58m), VJ, 1.5 mile (2.4km) run, and % drop were significant predictors of skating performance for repeat skate (slowest, fastest, and average time) and 44.80 m speed time, respectively. Four predictive equations were derived from multiple regression analyses: 1) slowest repeat skate time = 2.362 + (1.68 x 40-yd dash time) + (0.005 x 1.5 mile run), 2) fastest repeat skate time = 9.762 - (0.089 x VJ) - (0.998 x 40-yd dash time), 3) average repeat skate time = 7.770 + (1.041 x 40-yd dash time) - (0.63 x VJ) + (0.003 x 1.5 mile time), and 4) 47.85 m speed test = 7.707 - (0.050 x VJ) - (0.01 x % drop). It was concluded that selected off-ice tests could be used to predict on-ice performance regarding speed and recovery ability in Division III male and female hockey players. Key pointsThe 40-yd dash (36.58m) and vertical jump tests are significant predictors of on-ice skating performance specific to speed.In addition to 40-yd dash and vertical jump, the 1.5 mile (2.4km) run for time and percent power drop from the Wingate anaerobic power test were also significant predictors of skating performance that incorporates the aspect of recovery from skating activity.Due to the specificity of selected off-ice variables as predictors of on-ice performance, coaches can elect to assess player performance off-ice and focus on other uses of valuable ice time for their individual teams.

The selective activation of a C-F bond with an auxiliary strong Lewis acid: a method to change the activation preference of C-F and C-H bonds.

PubMed

Wang, Lin; Sun, Hongjian; Li, Xiaoyan; Fuhr, Olaf; Fenske, Dieter

2016-11-15

The selective activation of the C-F bonds in substituted (2,6-difluorophenyl)phenylimines (2,6-F 2 H 3 C 6 -(C[double bond, length as m-dash]NH)-n'-R-C 6 H 4 (n' = 2, R = H (1); n' = 2, R = Me (2); n' = 4, R = tBu (3))) by Fe(PMe 3 ) 4 with an auxiliary strong Lewis acid (LiBr, LiI, or ZnCl 2 ) was explored. As a result, iron(ii) halides ((H 5 C 6 -(C[double bond, length as m-dash]NH)-2-FH 3 C 6 )FeX(PMe 3 ) 3 (X = Br (8); Cl (9)) and (n-RH 4 C 6 -(C[double bond, length as m-dash]NH)-2'-FH 3 C 6 )FeX(PMe 3 ) 3 (n = 2, R = Me, X = Br (11); n = 4, R = tBu, X = I (12))) were obtained. Under similar reaction conditions, using LiBF 4 instead of LiBr or ZnCl 2 , the reaction of (2,6-difluorophenyl)phenylimine with Fe(PMe 3 ) 4 afforded an ionic complex [(2,6-F 2 H 3 C 6 -(C[double bond, length as m-dash]NH)-H 4 C 6 )Fe(PMe 3 ) 4 ](BF 4 ) (10) via the activation of a C-H bond. The method of C-F bond activation with an auxiliary strong Lewis acid is appropriate for monofluoroarylmethanimines. Without the Lewis acid, iron(ii) hydrides ((2-RH 4 C 6 -(C[double bond, length as m-dash]NH)-2'-FH 3 C 6 )FeH(PMe 3 ) 3 (R = H (13); Me (14))) were generated from the reactions of Fe(PMe 3 ) 4 with the monofluoroarylmethanimines (2-FH 4 C 6 -(C[double bond, length as m-dash]NH)-2'-RC 6 H 4 (R = H (4); Me (5))); however, in the presence of ZnCl 2 or LiBr, iron(ii) halides ((2-RH 4 C 6 -(C[double bond, length as m-dash]NH)-H 4 C 6 )FeX(PMe 3 ) 3 (R = H, X = Cl (15); R = Me, X = Br (16))) could be obtained through the activation of a C-F bond. Furthermore, a C-F bond activation with good regioselectivity in (pentafluorophenyl)arylmethanimines (F 5 C 6 -(C[double bond, length as m-dash]NH)-2,6-Y 2 C 6 H 3 (Y = F (6); H (7))) could be realized in the presence of ZnCl 2 to produce iron(ii) chlorides ((2,6-Y 2 H 3 C 6 -(C[double bond, length as m-dash]NH)-F 4 C 6 )FeCl(PMe 3 ) 3 (Y = F (17); H (18))). This series of iron(ii) halides could be used to catalyze the hydrosilylation reaction of aldehydes. Due to the stability of iron(ii) halides to high temperature, the reaction mixture was allowed to be heated to 100 °C and the reaction could finish within 0.5 h.

Proteinase pattern of honeybee prepupae from healthy and American Foulbrood infected bees investigated by zymography.

PubMed

Felicioli, Antonio; Turchi, Barbara; Fratini, Filippo; Giusti, Matteo; Nuvoloni, Roberta; Dani, Francesca Romana; Sagona, Simona

2018-05-15

American foulbrood disease (AFB) is the main devastating disease that affects honeybees' brood, caused by Paenibacillus larvae. The trend of the research on AFB has addressed the mechanisms by which P. larvae bacteria kill honeybee larvae. Since prepupae could react to the infection of AFB by increasing protease synthesis, the aim of this work was to compare protease activity in worker prepupae belonging to healthy colonies and to colonies affected by AFB. This investigation was performed by zymography. In gel, proteolytic activity was observed in prepupae extracts belonging only to the healthy colonies. In the prepupae extracts, 2D zimography followed by protein identification by MS allowed to detect Trypsin-1 and Chymotrypsin-1, which were not observed in diseased specimens. Further investigations are needed to clarify the involvement of these proteinases in the immune response of honeybee larvae and the mechanisms by which P. larvae inhibits protease production in its host. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Proteases in cardiometabolic diseases: Pathophysiology, molecular mechanisms and clinical applications

PubMed Central

Hua, Yinan; Nair, Sreejayan

2014-01-01

Cardiovascular disease is the leading cause of death in the U.S. and other developed country. Metabolic syndrome, including obesity, diabetes/insulin resistance, hypertension and dyslipidemia is major threat for public health in the modern society. It is well established that metabolic syndrome contributes to the development of cardiovascular disease collective called as cardiometabolic disease. Despite documented studies in the research field of cardiometabolic disease, the underlying mechanisms are far from clear. Proteases are enzymes that break down proteins, many of which have been implicated in various diseases including cardiac disease. Matrix metalloproteinase (MMP), calpain, cathepsin and caspase are among the major proteases involved in cardiac remodeling. Recent studies have also implicated proteases in the pathogenesis of cardiometabolic disease. Elevated expression and activities of proteases in atherosclerosis, coronary heart disease, obesity/insulin-associated heart disease as well as hypertensive heart disease have been documented. Furthermore, transgenic animals that are deficient in or overexpress proteases allow scientists to understand the causal relationship between proteases and cardiometabolic disease. Mechanistically, MMPs and cathepsins exert their effect on cardiometabolic diseases mainly through modifying the extracellular matrix. However, MMP and cathepsin are also reported to affect intracellular proteins, by which they contribute to the development of cardiometabolic diseases. On the other hand, activation of calpain and caspases has been shown to influence intracellular signaling cascade including the NF-κB and apoptosis pathways. Clinically, proteases are reported to function as biomarkers of cardiometabolic diseases. More importantly, the inhibitors of proteases are credited with beneficial cardiometabolic profile, although the exact molecular mechanisms underlying these salutary effects are still under investigation. A better understanding of the role of MMPs, cathepsins, calpains and caspases in cardiometabolic diseases process may yield novel therapeutic targets for threating or controlling these diseases. PMID:24815358

Dietary Fat, Eicosanoids and Breast Cancer Risk

DTIC Science & Technology

2005-04-01

Celery log 1Og log Peas 15g 15g 15g Garlic Powder/Salt/Pepper Dash Dash Dash Lemon Juice 2g 2g 2g Mayonnaise 25g 9g 0 Fat Free Miracle Whip 0 20g 30g...Sugar 0 4g 4g Oatmeal Walnut Cookie 30g 30g 30g Dinner: Beef Tenderloin 65g 60g 75g Mashed Potatoes 10Og 140g 140g w/Butter/Olive Oil 5g/4g 0 0 Corn 80g...Fat Free Gravy 40g 40g 40g w/ Safflower Oil/Flax Oil 7g (Safflower) 3g (Safflower) 3g (Flax) Mashed Potato 100g 140g 140g Green Beans 100g 100g �g

Influence of ensiling, exogenous protease addition, and bacterial inoculation on fermentation profile, nitrogen fractions, and ruminal in vitro starch digestibility in rehydrated and high-moisture corn.

PubMed

Ferraretto, L F; Fredin, S M; Shaver, R D

2015-10-01

Exogenous protease addition may be an option to increase proteolysis of zein proteins and thus starch digestibility in rehydrated and high-moisture corn (HMC) ensiled for short periods. In addition, microbial inoculation may accelerate fermentation and increase acid production and thus increase solubilization of zein proteins. Four experiments were performed to evaluate the effect on fermentation profile, N fractions, and ruminal in vitro starch digestibility (ivSD) of the following: (1) rehydration and ensiling of dry ground corn; (2) exogenous protease addition to rehydrated un-ensiled and ensiled corn; (3) exogenous protease addition or inoculation in rehydrated ensiled corn; and (4) exogenous protease addition or inoculation in HMC. Experiments 1, 2, and 3 were performed with 7 treatments: dry ground corn (DGC); DGC rehydrated to a targeted dry matter content of 70% (REH); REH treated with exogenous protease (REH+); REH ensiled for 30 d (ENS); ENS treated with exogenous protease (ENS+); ENS treated with a microbial inoculant containing Lactobacillus plantarum, Lactobacillus casei, Enterococcus faecium, and Pediococcus sp. (ENSI); and ENS treated with exogenous protease and microbial inoculant (ENSI+). Experiment 1 compared DGC, REH, and ENS with ivSD being greater for ENS (64.9%) than DGC and REH (51.7% on average). Experiment 2 compared REH and ENS without or with exogenous protease addition (REH+ and ENS+, respectively). Ensiling and exogenous protease addition increased ivSD, but exogenous protease addition was more effective in ENS than REH (6.4 vs. 2.6 percentage unit increase). Experiment 3 compared the effects of exogenous protease addition and inoculation in ENS corn (ENS, ENS+, ENSI, and ENSI+). The addition of protease, but not inoculant, increased ivSD. Inoculation reduced pH and acetate, propionate, and ethanol concentrations, and increased lactate and total acid concentrations. In experiment 4, 8 treatments were a combination of HMC noninoculated or inoculated with 1 of 3 microbial inoculants and with or without exogenous protease addition. The inoculant treatments contained (1) Lactobacillus buchneri 40788 and Pediococcus pentosaceus, (2) L. buchneri 40788, and (3) a mixture of P. pentosaceus and Propionibacterium freudenreichii. Protease, but not inoculation, increased ivSD by 7.5 percentage units (44.4 vs. 51.9%). Protease addition increased ivSD in rehydrated corn and HMC. Microbial inoculation improved fermentation profiles but did not affect ivSD. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Loss of second and sixth conserved cysteine residues from trypsin inhibitor-like cysteine-rich domain-type protease inhibitors in Bombyx mori may induce activity against microbial proteases.

PubMed

Li, Youshan; Liu, Huawei; Zhu, Rui; Xia, Qingyou; Zhao, Ping

2016-12-01

Previous studies have indicated that most trypsin inhibitor-like cysteine-rich domain (TIL)-type protease inhibitors, which contain a single TIL domain with ten conserved cysteines, inhibit cathepsin, trypsin, chymotrypsin, or elastase. Our recent findings suggest that Cys 2nd and Cys 6th were lost from the TIL domain of the fungal-resistance factors in Bombyx mori, BmSPI38 and BmSPI39, which inhibit microbial proteases and the germination of Beauveria bassiana conidia. To reveal the significance of these two missing cysteines in relation to the structure and function of TIL-type protease inhibitors in B. mori, cysteines were introduced at these two positions (D36 and L56 in BmSPI38, D38 and L58 in BmSPI39) by site-directed mutagenesis. The homology structure model of TIL domain of the wild-type and mutated form of BmSPI39 showed that two cysteine mutations may cause incorrect disulfide bond formation of B. mori TIL-type protease inhibitors. The results of Far-UV circular dichroism (CD) spectra indicated that both the wild-type and mutated form of BmSPI39 harbored predominantly random coil structures, and had slightly different secondary structure compositions. SDS-PAGE and Western blotting analysis showed that cysteine mutations affected the multimerization states and electrophoretic mobility of BmSPI38 and BmSPI39. Activity staining and protease inhibition assays showed that the introduction of cysteine mutations dramaticly reduced the activity of inhibitors against microbial proteases, such as subtilisin A from Bacillus licheniformis, protease K from Engyodontium album, protease from Aspergillus melleus. We also systematically analyzed the key residue sites, which may greatly influence the specificity and potency of TIL-type protease inhibitors. We found that the two missing cysteines in B. mori TIL-type protease inhibitors might be crucial for their inhibitory activities against microbial proteases. The genetic engineering of TIL-type protease inhibitors may be applied in both health care and agricultural industries, and could lead to new methods for breeding fungus-resistant transgenic crops and antifungal transgenic silkworm strains. Copyright © 2016 Elsevier Inc. All rights reserved.

Regulated Proteolysis in Bacteria.

PubMed

Mahmoud, Samar A; Chien, Peter

2018-06-20

Regulated proteolysis is a vital process that affects all living things. Bacteria use energy-dependent AAA+ proteases to power degradation of misfolded and native regulatory proteins. Given that proteolysis is an irreversible event, specificity and selectivity in degrading substrates are key. Specificity is often augmented through the use of adaptors that modify the inherent specificity of the proteolytic machinery. Regulated protein degradation is intricately linked to quality control, cell-cycle progression, and physiological transitions. In this review, we highlight recent work that has shed light on our understanding of regulated proteolysis in bacteria. We discuss the role AAA+ proteases play during balanced growth as well as how these proteases are deployed during changes in growth. We present examples of how protease selectivity can be controlled in increasingly complex ways. Finally, we describe how coupling a core recognition determinant to one or more modifying agents is a general theme for regulated protein degradation.

Glycosylation analysis of recombinant neutral protease I from Aspergillus oryzae expressed in Pichia pastoris.

PubMed

Lei, Da; Xu, Yang; He, Qinghua; Pang, Yifeng; Chen, Bo; Xiong, Liang; Li, Yanping

2013-12-01

Neutral protease I from Aspergillus oryzae 3.042 was expressed in Pichia pastoris and its N-glycosylation properties were analyzed. After purification by nickel-affinity chromatography column, the recombinant neutral protease (rNPI) was confirmed to be N-glycosylated by periodicacid/Schiff's base staining and Endo H digestion. Moreover, the deglycosylated protein's molecular weight decreased to 43.3 kDa from 54.5 kDa analyzed by SDS-PAGE and MALDI-TOF-MS, and the hyperglycosylation extent was 21 %. The N-glycosylation site of rNPI was analyzed by nano LC-MS/MS after digesting by trypsin and Glu-C, and the unique potential site Asn41 of mature peptide was found to be glycosylated. Homology modeling of the 3D structure of rNPI indicated that the attached N-glycans hardly affected neutral protease's activity due to the great distance away from the active site of the enzyme.

The protease-activated receptor-2 upregulates keratinocyte phagocytosis.

PubMed

Sharlow, E R; Paine, C S; Babiarz, L; Eisinger, M; Shapiro, S; Seiberg, M

2000-09-01

The protease-activated receptor-2 (PAR-2) belongs to the family of seven transmembrane domain receptors, which are activated by the specific enzymatic cleavage of their extracellular amino termini. Synthetic peptides corresponding to the tethered ligand domain (SLIGRL in mouse, SLIGKV in human) can activate PAR-2 without the need for receptor cleavage. PAR-2 activation is involved in cell growth, differentiation and inflammatory processes, and was shown to affect melanin and melanosome ingestion by human keratinocytes. Data presented here suggest that PAR-2 activation may regulate human keratinocyte phagocytosis. PAR-2 activation by trypsin, SLIGRL or SLIGKV increased the ability of keratinocytes to ingest fluorescently labeled microspheres or E. coli K-12 bioparticles. This PAR-2 mediated increase in keratinocyte phagocytic capability correlated with an increase in actin polymerization and *-actinin reorganization, cell surface morphological changes and increased soluble protease activity. Moreover, addition of serine protease inhibitors downmodulated both the constitutive and the PAR-2 mediated increases in phagocytosis, suggesting that serine proteases mediate this functional activity in keratinocytes. PAR-2 involvement in keratinocyte phagocytosis is a novel function for this receptor.

Influence of nitrogen source and pH value on undesired poly(γ-glutamic acid) formation of a protease producing Bacillus licheniformis strain.

PubMed

Meissner, Lena; Kauffmann, Kira; Wengeler, Timo; Mitsunaga, Hitoshi; Fukusaki, Eiichiro; Büchs, Jochen

2015-09-01

Bacillus spp. are used for the production of industrial enzymes but are also known to be capable of producing biopolymers such as poly(γ-glutamic acid). Biopolymers increase the viscosity of the fermentation broth, thereby impairing mixing, gas/liquid mass and heat transfer in any bioreactor system. Undesired biopolymer formation has a significant impact on the fermentation and downstream processing performance. This study shows how undesirable poly(γ-glutamic acid) formation of an industrial protease producing Bacillus licheniformis strain was prevented by switching the nitrogen source from ammonium to nitrate. The viscosity was reduced from 32 to 2.5 mPa s. A constant or changing pH value did not influence the poly(γ-glutamic acid) production. Protease production was not affected: protease activities of 38 and 46 U mL(-1) were obtained for ammonium and nitrate, respectively. With the presented results, protease production with industrial Bacillus strains is now possible without the negative impact on fermentation and downstream processing by undesired poly(γ-glutamic acid) formation.

Examination of the pronator quadratus muscle during hardware removal procedures after volar plating for distal radius fractures.

PubMed

Nho, Jae-Hwi; Gong, Hyun Sik; Song, Cheol Ho; Wi, Seung Myung; Lee, Young Ho; Baek, Goo Hyun

2014-09-01

It is not clear whether the pronator quadratus (PQ) muscle actually heals and provides a meaningful pronation force after volar plating for distal radius fractures (DRFs). We aimed to determine whether the length of the PQ muscle, which is dissected and then repaired during volar plating for a DRF, affects the forearm rotation strength and clinical outcomes. We examined 41 patients who requested hardware removal after volar plating. We measured the isokinetic forearm rotation strength and clinical outcomes including grip strength, wrist range of motion, and disabilities of the arm, shoulder and hand (DASH) scores at 6 months after fracture fixation. During the hardware removal surgery, which was performed at an average of 9 months (range, 8.3 to 11.5 months) after fracture fixation, we measured the PQ muscle length. The average PQ muscle length was 68% of the normal muscle length, and no significant relationship was found between the PQ muscle length and the outcomes including isokinetic forearm rotation strength, grip strength, wrist range of motion, and DASH scores. This study demonstrates that the length of the healed PQ muscle does not affect isokinetic forearm rotation strength and clinical outcomes after volar plating for DRFs. The results of this study support our current practice of loose repair of the PQ that is performed by most of the surgeons to prevent tendon irritation over the plate, and suggest that tight repair of the PQ is not necessary for achieving improved forearm function.

Pyoverdine and proteases affect the response of Pseudomonas aeruginosa to gallium in human serum.

PubMed

Bonchi, Carlo; Frangipani, Emanuela; Imperi, Francesco; Visca, Paolo

2015-09-01

Gallium is an iron mimetic which has recently been repurposed as an antibacterial agent due to its capability to disrupt bacterial iron metabolism. In this study, the antibacterial activity of gallium nitrate [Ga(NO3)3] was investigated in complement-free human serum (HS) on 55 Pseudomonas aeruginosa clinical isolates from cystic fibrosis and non-cystic fibrosis patients. The susceptibility of P. aeruginosa to Ga(NO3)3 in HS was dependent on the bacterial ability to acquire iron from serum binding proteins (i.e., transferrin). The extent of serum protein degradation correlated well with P. aeruginosa growth in HS, while pyoverdine production did not. However, pyoverdine-deficient P. aeruginosa strains were unable to grow in HS and overcome iron restriction, albeit capable of releasing proteases. Predigestion of HS with proteinase K promoted the growth of all strains, irrespective of their ability to produce proteases and/or pyoverdine. The MICs of Ga(NO3)3 were higher in HS than in an iron-poor Casamino Acids medium, where proteolysis does not affect iron availability. Coherently, strains displaying high proteolytic activity were less susceptible to Ga(NO3)3 in HS. Our data support a model in which both pyoverdine and proteases affect the response of P. aeruginosa to Ga(NO3)3 in HS. The relatively high Ga(NO3)3 concentration required to inhibit the growth of highly proteolytic P. aeruginosa isolates in HS poses a limitation to the potential of Ga(NO3)3 in the treatment of P. aeruginosa bloodstream infections. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Pyoverdine and Proteases Affect the Response of Pseudomonas aeruginosa to Gallium in Human Serum

PubMed Central

Bonchi, Carlo; Frangipani, Emanuela; Imperi, Francesco

2015-01-01

Gallium is an iron mimetic which has recently been repurposed as an antibacterial agent due to its capability to disrupt bacterial iron metabolism. In this study, the antibacterial activity of gallium nitrate [Ga(NO3)3] was investigated in complement-free human serum (HS) on 55 Pseudomonas aeruginosa clinical isolates from cystic fibrosis and non-cystic fibrosis patients. The susceptibility of P. aeruginosa to Ga(NO3)3 in HS was dependent on the bacterial ability to acquire iron from serum binding proteins (i.e., transferrin). The extent of serum protein degradation correlated well with P. aeruginosa growth in HS, while pyoverdine production did not. However, pyoverdine-deficient P. aeruginosa strains were unable to grow in HS and overcome iron restriction, albeit capable of releasing proteases. Predigestion of HS with proteinase K promoted the growth of all strains, irrespective of their ability to produce proteases and/or pyoverdine. The MICs of Ga(NO3)3 were higher in HS than in an iron-poor Casamino Acids medium, where proteolysis does not affect iron availability. Coherently, strains displaying high proteolytic activity were less susceptible to Ga(NO3)3 in HS. Our data support a model in which both pyoverdine and proteases affect the response of P. aeruginosa to Ga(NO3)3 in HS. The relatively high Ga(NO3)3 concentration required to inhibit the growth of highly proteolytic P. aeruginosa isolates in HS poses a limitation to the potential of Ga(NO3)3 in the treatment of P. aeruginosa bloodstream infections. PMID:26149986

Serpin-9 and -13 regulate hemolymph proteases during immune responses of Manduca sexta.

PubMed

He, Yan; Wang, Yang; Zhao, Picheng; Rayaprolu, Subrahmanyam; Wang, Xiuhong; Cao, Xiaolong; Jiang, Haobo

2017-11-01

Serpins are a superfamily of proteins, most of which inhibit cognate serine proteases by forming inactive acyl-enzyme complexes. In the tobacco hornworm Manduca sexta, serpin-1, -3 through -7 negatively regulate a hemolymph serine protease system that activates precursors of the serine protease homologs (SPHs), phenoloxidases (POs), Spätzles, and other cytokines. Here we report the cloning and characterization of M. sexta serpin-9 and -13. Serpin-9, a 402-residue protein most similar to Drosophila Spn77Ba, has R 366 at the P1 position right before the cleavage site; Serpin-13, a 444-residue ortholog of Drosophila Spn28Dc, is longer than the other seven serpins and has R 410 as the P1 residue. Both serpins are mainly produced in fat body and secreted into plasma to function. While their mRNA and protein levels were not up-regulated upon immune challenge, they blocked protease activities and affected proPO activation in hemolymph. Serpin-9 inhibited human neutrophil elastase, cathepsin G, trypsin, and chymotrypsin to different extents; serpin-13 reduced trypsin activity to approximately 10% at a molar ratio of 4:1 (serpin: enzyme). Serpin-9 was cleaved at Arg 366 by the enzymes with different specificity, but serpin-13 had four P1 sites (Arg 410 for trypsin-like proteases, Gly 406 and Ala 409 for the elastase and Thr 404 for cathepsin G). Supplementation of induced cell-free hemolymph (IP, P for plasma) with recombinant serpin-9 did not noticeably affect proPO activation, but slightly reduced the PO activity increase after 0-50% ammonium sulfate fraction of the IP had been elicited by bacteria. In comparison, addition of recombinant serpin-13 significantly inhibited proPO activation in IP and the suppression was stronger in the fraction of IP. Serpin-9- and -13-containing protein complexes were isolated from IP using their antibodies. Hemolymph protease-1 precursor (proHP1), HP6 and HP8 were found to be associated with serpin-9, whereas proHP1, HP2 and HP6 were pulled downed with serpin-13. These results indicate that both serpins regulate immune proteases in hemolymph of M. sexta larvae. Copyright © 2017 Elsevier Ltd. All rights reserved.

Asymmetric oxidation of vinyl- and ethynyl terthiophene ligands in triruthenium complexes.

PubMed

Zhang, Jing; Sun, Chao-Fang; Zhang, Ming-Xing; Hartl, František; Yin, Jun; Yu, Guang-Ao; Rao, Li; Liu, Sheng Hua

2016-01-14

A series of ruthenium(ii) complexes [{RuCl(CO)(PMe3)3(-CH[double bond, length as m-dash]CH-)}nX], (: n = 3, X = 3,3''-dimethyl-2,2':3',2''-terthiophene; : n = 2, X = 2,2'-bithiophene; : n = 2, X = 2,3-bis(3-methylthiophen-2-yl)benzothiophene) and [{Cp*(dppe)2Ru(-C[triple bond, length as m-dash]C-)}3X], (X = 3,3''-dimethyl-2,2':3',2''-terthiophene), were prepared and characterized by (1)H, (13)C and (31)P NMR. Their redox, spectroscopic and bonding properties were studied with a range of spectro-electrochemical methods in combination with density functional theory calculations. The first two anodic steps observed for and are largely localized on the lateral frameworks of the molecular triangle, the direct conjugation between them being precluded due to the photostable open form of the dithienyl ethene moiety. The third anodic step is then mainly localized on the centerpiece of the triangular structure, affecting both bithiophene laterals. The experimental IR and UV-vis-NIR spectroelectrochemical data and, largely, also DFT calculations account for this explanation, being further supported by direct comparison with the anodic behavior of reference diruthenium complexes and .

Patterns of Emotional and Behavioural Disturbance Associated with Autistic Traits in Young People with Severe Intellectual Disabilities and Challenging Behaviours

ERIC Educational Resources Information Center

Hill, Jennie; Furniss, Frederick

2006-01-01

Emotional and behavioural disturbance was assessed in 82 individuals with severe intellectual disabilities and challenging behaviour using the Diagnostic Assessment for the Severely Handicapped-II (DASH-II). Levels of disturbance were compared firstly in individuals with and without features of autism as assessed by the DASH-II, and secondly in…

75 FR 9756 - Airworthiness Directives; Bombardier, Inc. Model DHC-8-100 and DHC-8-200 Series Airplanes, and...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-04

... defined in Part 1 of the Bombardier (de Havilland) DHC-6 Twin Otter, Dash 7 & Dash 8 Corrosion Prevention... Corrosion Prevention and Control Manual PSM 1-GEN- 5, Part 1, Revision 3, contains the revision level of... more likely to exhibit indications of corrosion. We are issuing this AD to prevent structural failure...

Validation of Core Temperature Estimation Algorithm

DTIC Science & Technology

2016-01-29

plot of observed versus estimated core temperature with the line of identity (dashed) and the least squares regression line (solid) and line equation...estimated PSI with the line of identity (dashed) and the least squares regression line (solid) and line equation in the top left corner. (b) Bland...for comparison. The root mean squared error (RMSE) was also computed, as given by Equation 2.

Accuracy of a disability instrument to identify workers likely to develop upper extremity musculoskeletal disorders.

PubMed

Stover, Bert; Silverstein, Barbara; Wickizer, Thomas; Martin, Diane P; Kaufman, Joel

2007-06-01

Work related upper extremity musculoskeletal disorders (MSD) result in substantial disability, and expense. Identifying workers or jobs with high risk can trigger intervention before workers are injured or the condition worsens. We investigated a disability instrument, the QuickDASH, as a workplace screening tool to identify workers at high risk of developing upper extremity MSDs. Subjects included workers reporting recurring upper extremity MSD symptoms in the past 7 days (n = 559). The QuickDASH was reasonably accurate at baseline with sensitivity of 73% for MSD diagnosis, and 96% for symptom severity. Specificity was 56% for diagnosis, and 53% for symptom severity. At 1-year follow-up sensitivity and specificity for MSD diagnosis was 72% and 54%, respectively, as predicted by the baseline QuickDASH score. For symptom severity, sensitivity and specificity were 86% and 52%. An a priori target sensitivity of 70% and specificity of 50% was met by symptom severity, work pace and quality, and MSD diagnosis. The QuickDASH may be useful for identifying jobs or workers with increased risk for upper extremity MSDs. It may provide an efficient health surveillance screening tool useful for targeting early workplace intervention for prevention of upper extremity MSD problems.

System for line drawings interpretation

NASA Astrophysics Data System (ADS)

Boatto, L.; Consorti, Vincenzo; Del Buono, Monica; Eramo, Vincenzo; Esposito, Alessandra; Melcarne, F.; Meucci, Mario; Mosciatti, M.; Tucci, M.; Morelli, Arturo

1992-08-01

This paper describes an automatic system that extracts information from line drawings, in order to feed CAD or GIS systems. The line drawings that we analyze contain interconnected thin lines, dashed lines, text, and symbols. Characters and symbols may overlap with lines. Our approach is based on the properties of the run representation of a binary image that allow giving the image a graph structure. Using this graph structure, several algorithms have been designed to identify, directly in the raster image, straight segments, dashed lines, text, symbols, hatching lines, etc. Straight segments and dashed lines are converted into vectors, with high accuracy and good noise immunity. Characters and symbols are recognized by means of a recognizer, specifically developed for this application, designed to be insensitive to rotation and scaling. Subsequent processing steps include an `intelligent'' search through the graph in order to detect closed polygons, dashed lines, text strings, and other higher-level logical entities, followed by the identification of relationships (adjacency, inclusion, etc.) between them. Relationships are further translated into a formal description of the drawing. The output of the system can be used as input to a Geographic Information System package. The system is currently used by the Italian Land Register Authority to process cadastral maps.

The influence of education and income on responses to the QuickDASH questionnaire.

PubMed

Finsen, V

2015-05-01

We studied the influence of levels of income and education on QuickDASH scores. The scores were collected in a random sample of 1376 residents of Norway. The level of income was divided into four bands and level of education into five bands. The mean QuickDASH score for both men and women fell with every increase in education and income level. For women the mean score was 30 for those with the shortest education and 9 for those with the longest (p < 0.001). The corresponding figures for men were 19 and 7 (p < 0.01). The women with the lowest level of income had a mean score of 23, compared with 8 for women with the highest income level (p < 0.001). For men the corresponding mean scores were 20 and 5 (p < 0.001). Analysis of variance showed that age alone accounted for 16% of the variability of the scores among women and 7% among men. When levels of education and income were added to the analysis, these three factors accounted for 21% of the variability among women and 13% among men. We conclude that socioeconomic factors significantly influence QuickDASH scores. 3. © The Author(s) 2014.

Regulation of protein degradation in muscle by calcium

NASA Technical Reports Server (NTRS)

Zeman, Richard J.; Kameyama, Tsuneo; Matsumoto, Kazue; Bernstein, Paul; Etlinger, Joseph D.

1985-01-01

Calcium-dependent regulation of intracellular protein degradation was studied in isolated rat skeletal muscles incubated in vitro in the presence of a large variety of agents known to affect calcium movement and distribution. The effect of different classes of protease inhibitors was tested to determine the responsible proteolytic systems involved in calcium-dependent degradation. The results suggest that nonlysosomal leupetin- and E-64-c-sensitive proteases are resposible for calcium-dependent proteolysis in muscle.

Effects of exogenous proteases without or with carbohydrases on nutrient digestibility and disappearance of non-starch polysaccharides in broiler chickens

PubMed Central

Olukosi, O. A.; Beeson, L. A.; Englyst, K.; Romero, L. F.

2015-01-01

The objective of the current study was to evaluate the effect of a subtilisin protease, without or with inclusion of carbohydrases, on digestibility and retention of energy and protein, as well as the solubilization and disappearance of non-starch polysaccharides (NSP) from corn-soybean meal based diets fed to broiler chickens. Two hundred eighty-eight Ross 308 male broiler chickens were used for the experiment. On d 14, the birds were weighed and allocated to 6 treatments and 8 replicates per treatment with 6 birds per replicate. Treatments were: 1) corn-soybean meal based control diet; 2) control diet plus supplemental protease at 5,000 (P5000) protease units (PU)/kg); 3) control plus 10,000 PU/kg protease (P10000); or control plus an enzyme combination containing xylanase, amylase, and protease (XAP) added to achieve protease activity of: 4) 2,500 PU/kg (XAP2500); 5) 5,000 PU/kg (XAP5000); or 6) 10,000 PU/kg (XAP10000). The enzymes in XAP were combined at fixed ratios of 10:1:25 of xylanase:amylase:protease. Data were analyzed by ANOVA and specific orthogonal contrasts between treatments were performed. Addition of xylanase and amylase increased (P < 0.05) the ileal digestibility of protein by 4.2% and 2.1% at XAP5000 and XAP10000, respectively (relative to P5000 and P10000, respectively), exhibiting a plateau after the XAP5000 dose. Increment (P < 0.05) in AME due to protease was evident, particularly in P10000. At the ileal level, XAP reduced (P < 0.05) the flow of insoluble xylose and arabinose, which indicates an increase in the solubilization of arabinoxylan polymers in the small intestine. Protease on its own reduced (P < 0.05) the flow of insoluble arabinose but did not affect the flow of insoluble xylose. XAP reduced (P < 0.05) the pre-cecal flow of insoluble and total glucose and galactose. It was concluded that whereas protease by itself improved nutrient utilization and increased solubilization of NSP components, at the lower dose, a combination of xylanase, amylase, and protease produced effects greater than those of protease alone. PMID:26371327

Optimal symmetric flight studies

NASA Technical Reports Server (NTRS)

Weston, A. R.; Menon, P. K. A.; Bilimoria, K. D.; Cliff, E. M.; Kelley, H. J.

1985-01-01

Several topics in optimal symmetric flight of airbreathing vehicles are examined. In one study, an approximation scheme designed for onboard real-time energy management of climb-dash is developed and calculations for a high-performance aircraft presented. In another, a vehicle model intermediate in complexity between energy and point-mass models is explored and some quirks in optimal flight characteristics peculiar to the model uncovered. In yet another study, energy-modelling procedures are re-examined with a view to stretching the range of validity of zeroth-order approximation by special choice of state variables. In a final study, time-fuel tradeoffs in cruise-dash are examined for the consequences of nonconvexities appearing in the classical steady cruise-dash model. Two appendices provide retrospective looks at two early publications on energy modelling and related optimal control theory.

Successfully recruiting a multicultural population: the DASH-Sodium experience.

PubMed

Kennedy, Betty M; Conlin, Paul R; Ernst, Denise; Reams, Patrice; Charleston, Jeanne B; Appel, Lawrence J

2005-01-01

Recruiting practices employed by the four clinical centers participating in the Dietary Approaches to Stop Hypertension (DASH)-Sodium trial were examined to assess the most successful method of obtaining participants and to describe pertinent learning experiences gained as a result of the trial. The primary recruitment strategies employed by each center were mass mailing brochures (direct, coupon packs, or other) and mass media (advertisements in newspapers, radio, and television spots). Of 412 randomized participants, 265 (64%) were from mass distribution of brochures, 62 (15%) mass media, and 85 (21%) were prior study participants, referred by word-of-mouth, or reported coming from screening events and presentations. Although the most successful method of recruitment was mass mailing brochures, three times as many brochures were distributed to obtain similar success as in the initial DASH trial.

An on-board near-optimal climb-dash energy management

NASA Technical Reports Server (NTRS)

Weston, A. R.; Cliff, E. M.; Kelley, H. J.

1982-01-01

On-board real time flight control is studied in order to develop algorithms which are simple enough to be used in practice, for a variety of missions involving three dimensional flight. The intercept mission in symmetric flight is emphasized. Extensive computation is required on the ground prior to the mission but the ensuing on-board exploitation is extremely simple. The scheme takes advantage of the boundary layer structure common in singular perturbations, arising with the multiple time scales appropriate to aircraft dynamics. Energy modelling of aircraft is used as the starting point for the analysis. In the symmetric case, a nominal path is generated which fairs into the dash or cruise state. Feedback coefficients are found as functions of the remaining energy to go (dash energy less current energy) along the nominal path.

Altered Substrate Specificity of Drug-Resistant Human Immunodeficiency Virus Type 1 Protease

PubMed Central

Dauber, Deborah S.; Ziermann, Rainer; Parkin, Neil; Maly, Dustin J.; Mahrus, Sami; Harris, Jennifer L.; Ellman, Jon A.; Petropoulos, Christos; Craik, Charles S.

2002-01-01

Resistance to human immunodeficiency virus type 1 protease (HIV PR) inhibitors results primarily from the selection of multiple mutations in the protease region. Because many of these mutations are selected for the ability to decrease inhibitor binding in the active site, they also affect substrate binding and potentially substrate specificity. This work investigates the substrate specificity of a panel of clinically derived protease inhibitor-resistant HIV PR variants. To compare protease specificity, we have used positional-scanning, synthetic combinatorial peptide libraries as well as a select number of individual substrates. The subsite preferences of wild-type HIV PR determined by using the substrate libraries are consistent with prior reports, validating the use of these libraries to compare specificity among a panel of HIV PR variants. Five out of seven protease variants demonstrated subtle differences in specificity that may have significant impacts on their abilities to function in viral maturation. Of these, four variants demonstrated up to fourfold changes in the preference for valine relative to alanine at position P2 when tested on individual peptide substrates. This change correlated with a common mutation in the viral NC/p1 cleavage site. These mutations may represent a mechanism by which severely compromised, drug-resistant viral strains can increase fitness levels. Understanding the altered substrate specificity of drug-resistant HIV PR should be valuable in the design of future generations of protease inhibitors as well as in elucidating the molecular basis of regulation of proteolysis in HIV. PMID:11773410

Multiple Off-Ice Performance Variables Predict On-Ice Skating Performance in Male and Female Division III Ice Hockey Players

PubMed Central

Janot, Jeffrey M.; Beltz, Nicholas M.; Dalleck, Lance D.

2015-01-01

The purpose of this study was to determine if off-ice performance variables could predict on-ice skating performance in Division III collegiate hockey players. Both men (n = 15) and women (n = 11) hockey players (age = 20.5 ± 1.4 years) participated in the study. The skating tests were agility cornering S-turn, 6.10 m acceleration, 44.80 m speed, modified repeat skate, and 15.20 m full speed. Off-ice variables assessed were years of playing experience, height, weight and percent body fat and off-ice performance variables included vertical jump (VJ), 40-yd dash (36.58m), 1-RM squat, pro-agility, Wingate peak power and peak power percentage drop (% drop), and 1.5 mile (2.4km) run. Results indicated that 40-yd dash (36.58m), VJ, 1.5 mile (2.4km) run, and % drop were significant predictors of skating performance for repeat skate (slowest, fastest, and average time) and 44.80 m speed time, respectively. Four predictive equations were derived from multiple regression analyses: 1) slowest repeat skate time = 2.362 + (1.68 x 40-yd dash time) + (0.005 x 1.5 mile run), 2) fastest repeat skate time = 9.762 - (0.089 x VJ) - (0.998 x 40-yd dash time), 3) average repeat skate time = 7.770 + (1.041 x 40-yd dash time) - (0.63 x VJ) + (0.003 x 1.5 mile time), and 4) 47.85 m speed test = 7.707 - (0.050 x VJ) - (0.01 x % drop). It was concluded that selected off-ice tests could be used to predict on-ice performance regarding speed and recovery ability in Division III male and female hockey players. Key points The 40-yd dash (36.58m) and vertical jump tests are significant predictors of on-ice skating performance specific to speed. In addition to 40-yd dash and vertical jump, the 1.5 mile (2.4km) run for time and percent power drop from the Wingate anaerobic power test were also significant predictors of skating performance that incorporates the aspect of recovery from skating activity. Due to the specificity of selected off-ice variables as predictors of on-ice performance, coaches can elect to assess player performance off-ice and focus on other uses of valuable ice time for their individual teams. PMID:26336338

The Strategic Nature of the Tactical Satellite. Part 2

DTIC Science & Technology

2006-08-13

hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and...constellation orbiting at 100 NM. The curves represent data for three mission types: SIGINT (solid), comm/BFT (dashed), and imagery (dotted). Two panes...persistent constellation orbiting at 500 km. The curves represent data for three mission types: SIGINT (solid), comm/BFT (dashed), and imagery (dotted

DASH (Dietary Approaches to Stop Hypertension) Diet and Risk of Subsequent Kidney Disease

PubMed Central

Rebholz, Casey M.; Crews, Deidra C.; Grams, Morgan E.; Steffen, Lyn M.; Levey, Andrew S.; Miller, Edgar R.; Appel, Lawrence J.; Coresh, Josef

2016-01-01

Background There are established guidelines for recommended dietary intake for hypertension treatment and cardiovascular disease prevention. Evidence is lacking for effective dietary patterns for kidney disease prevention. Study Design Prospective cohort study Setting & Participants Atherosclerosis Risk in Communities (ARIC) study participants with baseline estimated glomerular filtration rate (eGFR) ≥60 mL/min/1.73 m2 (N=14,882) Predictor The Dietary Approaches to Stop Hypertension (DASH) diet score was calculated based on self-reported dietary intake of red and processed meat, sweetened beverages, sodium, fruits, vegetables, whole grains, nuts and legumes, and low-fat dairy products, averaged over two visits. Outcomes Cases were ascertained based on development of eGFR <60 mL/min/1.73 m2 accompanied by ≥25% eGFR decline from baseline, an ICD-9/10 code for a kidney disease–related hospitalization or death, or end-stage renal disease from baseline through 2012. Results A total of 3,720 participants developed kidney disease during a median follow-up of 23 years. Participants with a DASH diet score in the lowest tertile were 16% more likely to develop kidney disease than those with the highest score tertile (HR, 1.16; 95% CI, 1.07-1.26; p for trend <0.001), after adjusting for socio-demographics, smoking status, physical activity, total caloric intake, baseline eGFR, overweight/obese status, diabetes status, hypertension status, systolic blood pressure, and anti-hypertensive medication use. Of the individual components of the DASH diet score, high intake of red and processed meat was adversely associated with kidney disease and high intake of nuts, legumes, and low-fat dairy products was associated with reduced risk of kidney disease. Limitations Potential measurement error due to self-reported dietary intake and lack of data on albuminuria Conclusions Consuming a DASH-style diet was associated with lower risk for kidney disease, independent of demographic characteristics, established kidney risk factors, and baseline kidney function. Healthful dietary patterns, such as the DASH diet, may be beneficial for kidney disease prevention. PMID:27519166

DASH (Dietary Approaches to Stop Hypertension) Diet and Risk of Subsequent Kidney Disease.

PubMed

Rebholz, Casey M; Crews, Deidra C; Grams, Morgan E; Steffen, Lyn M; Levey, Andrew S; Miller, Edgar R; Appel, Lawrence J; Coresh, Josef

2016-12-01

There are established guidelines for recommended dietary intake for hypertension treatment and cardiovascular disease prevention. Evidence is lacking for effective dietary patterns for kidney disease prevention. Prospective cohort study. Atherosclerosis Risk in Communities (ARIC) Study participants with baseline estimated glomerular filtration rate (eGFR) ≥ 60mL/min/1.73m 2 (N=14,882). The Dietary Approaches to Stop Hypertension (DASH) diet score was calculated based on self-reported dietary intake of red and processed meat, sweetened beverages, sodium, fruits, vegetables, whole grains, nuts and legumes, and low-fat dairy products, averaged over 2 visits. Cases were ascertained based on the development of eGFRs<60mL/min/1.73m 2 accompanied by ≥25% eGFR decline from baseline, an International Classification of Diseases, Ninth/Tenth Revision code for a kidney disease-related hospitalization or death, or end-stage renal disease from baseline through 2012. 3,720 participants developed kidney disease during a median follow-up of 23 years. Participants with a DASH diet score in the lowest tertile were 16% more likely to develop kidney disease than those with the highest score tertile (HR, 1.16; 95% CI, 1.07-1.26; P for trend < 0.001), after adjusting for sociodemographics, smoking status, physical activity, total caloric intake, baseline eGFR, overweight/obese status, diabetes status, hypertension status, systolic blood pressure, and antihypertensive medication use. Of the individual components of the DASH diet score, high red and processed meat intake was adversely associated with kidney disease and high nuts, legumes, and low-fat dairy products intake was associated with reduced risk for kidney disease. Potential measurement error due to self-reported dietary intake and lack of data for albuminuria. Consuming a DASH-style diet was associated with lower risk for kidney disease independent of demographic characteristics, established kidney risk factors, and baseline kidney function. Healthful dietary patterns such as the DASH diet may be beneficial for kidney disease prevention. Copyright © 2016 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

DASH for asthma: A pilot study of the DASH diet in not-well-controlled adult asthma⋆

PubMed Central

Ma, Jun; Strub, Peg; Lavori, Phillip W.; Buist, A. Sonia; Camargo, Carlos A.; Nadeau, Kari C.; Wilson, Sandra R.; Xiao, Lan

2014-01-01

This pilot study aims to provide effect size confidence intervals, clinical trial and intervention feasibility data, and procedural materials for a full-scale randomized controlled trial that will determine the efficacy of Dietary Approaches to Stop Hypertension (DASH) as adjunct therapy to standard care for adults with uncontrolled asthma. The DASH diet encompasses foods (e.g., fresh fruit, vegetables, and nuts) and antioxidant nutrients (e.g., vitamins A, C, E, and zinc) with potential benefits for persons with asthma, but it is unknown whether the whole diet is beneficial. Participants (n = 90) will be randomized to receive usual care alone or combined with a DASH intervention consisting of 8 group and 3 individual sessions during the first 3 months, followed by at least monthly phone consultations for another 3 months. Follow-up assessments will occur at 3 and 6 months. The primary outcome measure is the 7-item Juniper Asthma Control Questionnaire, a validated composite measure of daytime and nocturnal symptoms, activity limitations, rescue medication use, and percentage predicted forced expiratory volume in 1 second. We will explore changes in inflammatory markers important to asthma pathophysiology (e.g., fractional exhaled nitric oxide) and their potential to mediate the intervention effect on disease control. We will also conduct pre-specified subgroup analyses by genotype (e.g., polymorphisms on the glutathione S transferase gene) and phenotype (e.g., atopy, obesity). By evaluating a dietary pattern approach to improving asthma control, this study could advance the evidence base for refining clinical guidelines and public health recommendations regarding the role of dietary modifications in asthma management. PMID:23648395

Standardized combined cryotherapy and compression using Cryo/Cuff after wrist arthroscopy.

PubMed

Meyer-Marcotty, M; Jungling, O; Vaske, B; Vogt, P M; Knobloch, Karsten

2011-02-01

cryotherapy and compression as integral part of the RICE regimen are thought to improve treatment outcome after sport injuries. Using standardized cryotherapy and compression perioperatively has been reported with conflicting clinical results. The impact of combined cryotherapy and compression is compared to standard care among patients undergoing wrist arthroscopy. fifty-six patients undergoing wrist arthroscopy were assessed, 54 patients were randomized to either Cryo/Cuff (3 × 10 min twice daily) or standard care over 3 weeks. Follow-up clinical visits were at postoperative days 1, 8, and 21. One patient in each group was lost during follow-up. Fifty-two patients were analyzed. Statistics were performed as Intention-to-treat analysis. Outcome parameters were pain, three-dimensional volume of the wrist, range of motion, and DASH score. the Cryo/Cuffgroup had a 49% reduction in pain level (VAS 3.5 ± 0.4 vs. VAS 1.8 ± 0.2 on the 21st postoperative day) when compared to a reduction of 41% in the control group (VAS 5.1 ± 0.6 preoperatively vs. VAS 3.0 ± 0.5 on the 21st postoperative day). Swelling and range of motion were not as significantly different between the two groups as were DASH scores (DASH-score Cryo/Cuff group preoperatively 37.3 ± 3.5 and postoperatively 36.9 ± 3.5; DASH-score control group preoperatively 42.8 ± 4.3 and postoperatively 41.9 ± 4.9). The CONSORT score reached 17 out of 22. there was no significant effect of additional home-based combined cryotherapy and compression using the Cryo/Cuff wrist bandage, following wrist arthroscopy regarding pain, swelling, range of motion, and subjective impairment assessed using the DASH score over 3 weeks in comparison with the control group.

Mediterranean and DASH Diet Scores and Mortality in Women with Heart Failure: The Women’s Health Initiative

PubMed Central

Levitan, Emily B.; Lewis, Cora E.; Tinker, Lesley F.; Eaton, Charles B.; Ahmed, Ali; Manson, JoAnn E.; Snetselaar, Linda G.; Martin, Lisa W.; Trevisan, Maurizio; Howard, Barbara V.; Shikany, James M.

2015-01-01

Background Current dietary recommendations for heart failure (HF) patients are largely based on data from non-HF populations; evidence regarding associations of dietary patterns with outcomes in HF is limited. We therefore evaluated associations of Mediterranean and DASH diet scores with mortality among postmenopausal women with HF. Methods and Results Women’s Health Initiative participants were followed from the date of HF hospitalization through the date of death or last participant contact prior to August 2009. Mediterranean and DASH diet scores were calculated from food-frequency questionnaires. Cox proportional hazards models adjusted for demographics, health behaviors, and health status were used to calculate hazard ratios (HR) and 95% confidence intervals (CI). Over a median of 4.6 years of follow-up, 1,385 of 3,215 (43.1%) participants who experienced a HF hospitalization died. Multivariable-adjusted HRs were 1 (reference), 1.05 (95% CI 0.89–1.24), 0.97 (95% CI 0.81–1.17), and 0.85 (95% CI 0.70–1.02) across quartiles of the Mediterranean diet score (p-trend = 0.08) and 1 (reference), 1.04 (95% CI 0.89–1.21), 0.83 (95% CI 0.70–0.98), and 0.84 (95% CI 0.70–1.00) across quartiles of the DASH diet score (p-trend = 0.01). Diet score components vegetables, must, and whole grain intake were inversely associated with mortality. Conclusions Higher DASH diet scores were associated with modestly lower mortality in women with HF, and there was a non-significant trend towards an inverse association with Mediterranean diet scores. These data provide support for the concept that dietary recommendations developed for other cardiovascular conditions or general populations may also be appropriate in HF patients. PMID:24107587

A priori-defined diet quality indices, biomarkers and risk for type 2 diabetes in five ethnic groups: the Multiethnic Cohort.

PubMed

Jacobs, Simone; Boushey, Carol J; Franke, Adrian A; Shvetsov, Yurii B; Monroe, Kristine R; Haiman, Christopher A; Kolonel, Laurence N; Le Marchand, Loic; Maskarinec, Gertraud

2017-08-01

Dietary indices have been related to risk for type 2 diabetes (T2D) predominantly in white populations. The present study evaluated this association in the ethnically diverse Multiethnic Cohort and examined four diet quality indices in relation to T2D risk, homoeostatic model assessment-estimated insulin resistance (HOMA-IR) and biomarkers of dyslipidaemia, inflammation and adipokines. The T2D analysis included 166 550 white, African American, Native Hawaiian, Japanese American and Latino participants (9200 incident T2D cases). Dietary intake was assessed at baseline using a quantitative FFQ and T2D status was based on three self-reports and confirmed by administrative data. Biomarkers were assessed about 10 years later in a biomarker subcohort (n 10 060). Sex- and ethnicity-specific hazard ratios were calculated for the Healthy Eating Index-2010 (HEI-2010), the alternative HEI-2010 (AHEI-2010), the alternate Mediterranean diet score (aMED) and the Dietary Approaches to Stop Hypertension (DASH). Multivariable-adjusted means of biomarkers were compared across dietary index tertiles in the biomarker subcohort. The AHEI-2010, aMED (in men only) and DASH scores were related to a 10-20 % lower T2D risk, with the strongest associations in whites and the direction of the relationships mostly consistent across ethnic groups. Higher scores on the four indices were related to lower HOMA-IR, TAG and C-reactive protein concentrations, not related to leptin, and the DASH score was directly associated with adiponectin. The AHEI-2010 and DASH were directly related to HDL-cholesterol in women. Potential underlying biological mechanisms linking diet quality and T2D risk are an improved lipid profile and reduced systemic inflammation and, with regards to DASH alone, an improved adiponectin profile.

MOLECULAR IDENTIFICATION OF CYSTEINE AND TRYPSIN PROTEASE, EFFECT OF DIFFERENT HOSTS ON PROTEASE EXPRESSION, AND RNAI MEDIATED SILENCING OF CYSTEINE PROTEASE GENE IN THE SUNN PEST.

PubMed

Amiri, Azam; Bandani, Ali Reza; Alizadeh, Houshang

2016-04-01

Sunn pest, Eurygaster integriceps, is a serious pest of cereals in the wide area of the globe from Near and Middle East to East and South Europe and North Africa. This study described for the first time, identification of E. integriceps trypsin serine protease and cathepsin-L cysteine, transcripts involved in digestion, which might serve as targets for pest control management. A total of 478 and 500 base pair long putative trypsin and cysteine gene sequences were characterized and named Tryp and Cys, respectively. In addition, the tissue-specific relative gene expression levels of these genes as well as gluten hydrolase (Gl) were determined under different host kernels feeding conditions. Result showed that mRNA expression of Cys, Tryp, and Gl was significantly affected after feeding on various host plant species. Transcript levels of these genes were most abundant in the wheat-fed E. integriceps larvae compared to other hosts. The Cys transcript was detected exclusively in the gut, whereas the Gl and Tryp transcripts were detectable in both salivary glands and gut. Also possibility of Sunn pest gene silencing was studied by topical application of cysteine double-stranded RNA (dsRNA). The results indicated that topically applied dsRNA on fifth nymphal stage can penetrate the cuticle of the insect and induce RNA interference. The Cys gene mRNA transcript in the gut was reduced to 83.8% 2 days posttreatment. Also, it was found that dsRNA of Cys gene affected fifth nymphal stage development suggesting the involvement of this protease in the insect growth, development, and molting. © 2015 Wiley Periodicals, Inc.

SERPINE1: A Molecular Switch in the Proliferation-Migration Dichotomy in Wound-“Activated” Keratinocytes

PubMed Central

Simone, Tessa M.; Higgins, Craig E.; Czekay, Ralf-Peter; Law, Brian K.; Higgins, Stephen P.; Archambeault, Jaclyn; Kutz, Stacie M.; Higgins, Paul J.

2014-01-01

Significance: A highly interactive serine protease/plasmin/matrix metalloproteinase axis regulates stromal remodeling in the wound microenvironment. Current findings highlight the importance of stringent controls on protease expression and their topographic activities in cell proliferation, migration, and tissue homeostasis. Targeting elements in this cascading network may lead to novel therapeutic approaches for fibrotic diseases and chronic wounds. Recent Advances: Matrix-active proteases and their inhibitors orchestrate wound site tissue remodeling, cell migration, and proliferation. Indeed, the serine proteases urokinase plasminogen activator and tissue-type plasminogen activator (uPA/tPA) and their major phsyiological inhibitor, plasminogen activator inhibitor-1 (PAI-1; serine protease inhibitor clade E member 1 [SERPINE1]), are upregulated in several cell types during injury repair. Coordinate expression of proteolytic enzymes and their inhibitors in the wound bed provides a mechanism for fine control of focal proteolysis to facilitate matrix restructuring and cell motility in complex environments. Critical Issues: Cosmetic and tissue functional consequences of wound repair anomalies affect the quality of life of millions of patients in the United States alone. The development of novel therapeutics to manage individuals most affected by healing anomalies will likely derive from the identification of critical, translationally accessible, control elements in the wound site microenvironment. Future Directions: Activation of the PAI-1 gene early after wounding, its prominence in the repair transcriptome and varied functions suggest a key role in the global cutaneous injury response program. Targeting PAI-1 gene expression and/or PAI-1 function with molecular genetic constructs, neutralizing antibodies or small molecule inhibitors may provide a novel, therapeutically relevant approach, to manage the pathophysiology of wound healing disorders associated with deficient or excessive PAI-1 levels. PMID:24669362

Arginine-specific gingipains from Porphyromonas gingivalis deprive protective functions of secretory leucocyte protease inhibitor in periodontal tissue

PubMed Central

Into, T; Inomata, M; Kanno, Y; Matsuyama, T; Machigashira, M; Izumi, Y; Imamura, T; Nakashima, M; Noguchi, T; Matsushita, K

2006-01-01

Chronic periodontitis is correlated with Porphyromonas gingivalis infection. In this study, we found that the expression of secretory leucocyte protease inhibitor (SLPI), an endogenous inhibitor for neutrophil-derived proteases, was reduced in gingival tissues with chronic periodontitis associated with P. gingivalis infection. The addition of vesicles of P. gingivalis decreased the amount of SLPI in the media of primary human gingival keratinocytes compared to untreated cultures. We therefore investigated how arginine-specific gingipains (Rgps) affect the functions of SLPI, because Rgps are the major virulence factors in the vesicles and cleave a wide range of in-host proteins. We found that Rgps digest SLPI in vitro, suppressing the release of SLPI. Rgps proteolysis of SLPI disrupted SLPI functions, which normally suppresses neutrophil elastase and neutralizes pro-inflammatory effects of bacterial cell wall compounds in cultured human gingival fibroblasts. The protease inhibitory action of SLPI was not exerted towards Rgps. These results suggest that Rgps reduce the protective effects of SLPI on neutrophil proteases and bacterial proinflammatory compounds, by which disease in gingival tissue may be accelerated at the sites with P. gingivalis infection. PMID:16907925

cl-dash: rapid configuration and deployment of Hadoop clusters for bioinformatics research in the cloud.

PubMed

Hodor, Paul; Chawla, Amandeep; Clark, Andrew; Neal, Lauren

2016-01-15

: One of the solutions proposed for addressing the challenge of the overwhelming abundance of genomic sequence and other biological data is the use of the Hadoop computing framework. Appropriate tools are needed to set up computational environments that facilitate research of novel bioinformatics methodology using Hadoop. Here, we present cl-dash, a complete starter kit for setting up such an environment. Configuring and deploying new Hadoop clusters can be done in minutes. Use of Amazon Web Services ensures no initial investment and minimal operation costs. Two sample bioinformatics applications help the researcher understand and learn the principles of implementing an algorithm using the MapReduce programming pattern. Source code is available at https://bitbucket.org/booz-allen-sci-comp-team/cl-dash.git. hodor_paul@bah.com. © The Author 2015. Published by Oxford University Press.

Road Lane Detection by Discriminating Dashed and Solid Road Lanes Using a Visible Light Camera Sensor.

PubMed

Hoang, Toan Minh; Hong, Hyung Gil; Vokhidov, Husan; Park, Kang Ryoung

2016-08-18

With the increasing need for road lane detection used in lane departure warning systems and autonomous vehicles, many studies have been conducted to turn road lane detection into a virtual assistant to improve driving safety and reduce car accidents. Most of the previous research approaches detect the central line of a road lane and not the accurate left and right boundaries of the lane. In addition, they do not discriminate between dashed and solid lanes when detecting the road lanes. However, this discrimination is necessary for the safety of autonomous vehicles and the safety of vehicles driven by human drivers. To overcome these problems, we propose a method for road lane detection that distinguishes between dashed and solid lanes. Experimental results with the Caltech open database showed that our method outperforms conventional methods.

Road Lane Detection by Discriminating Dashed and Solid Road Lanes Using a Visible Light Camera Sensor

PubMed Central

Hoang, Toan Minh; Hong, Hyung Gil; Vokhidov, Husan; Park, Kang Ryoung

2016-01-01

With the increasing need for road lane detection used in lane departure warning systems and autonomous vehicles, many studies have been conducted to turn road lane detection into a virtual assistant to improve driving safety and reduce car accidents. Most of the previous research approaches detect the central line of a road lane and not the accurate left and right boundaries of the lane. In addition, they do not discriminate between dashed and solid lanes when detecting the road lanes. However, this discrimination is necessary for the safety of autonomous vehicles and the safety of vehicles driven by human drivers. To overcome these problems, we propose a method for road lane detection that distinguishes between dashed and solid lanes. Experimental results with the Caltech open database showed that our method outperforms conventional methods. PMID:27548176

cl-dash: rapid configuration and deployment of Hadoop clusters for bioinformatics research in the cloud

PubMed Central

Hodor, Paul; Chawla, Amandeep; Clark, Andrew; Neal, Lauren

2016-01-01

Summary: One of the solutions proposed for addressing the challenge of the overwhelming abundance of genomic sequence and other biological data is the use of the Hadoop computing framework. Appropriate tools are needed to set up computational environments that facilitate research of novel bioinformatics methodology using Hadoop. Here, we present cl-dash, a complete starter kit for setting up such an environment. Configuring and deploying new Hadoop clusters can be done in minutes. Use of Amazon Web Services ensures no initial investment and minimal operation costs. Two sample bioinformatics applications help the researcher understand and learn the principles of implementing an algorithm using the MapReduce programming pattern. Availability and implementation: Source code is available at https://bitbucket.org/booz-allen-sci-comp-team/cl-dash.git. Contact: hodor_paul@bah.com PMID:26428290

Interspecific differences between D. pulex and D. magna in tolerance to cyanobacteria with protease inhibitors.

PubMed

Kuster, Christian J; Von Elert, Eric

2013-01-01

It is known that cyanobacteria negatively affect herbivores due to their production of toxins such as protease inhibitors. In the present study we investigated potential interspecific differences between two major herbivores, Daphnia magna and Daphnia pulex, in terms of their tolerance to cyanobacteria with protease inhibitors. Seven clones each of D. magna and of D. pulex were isolated from different habitats in Europe and North America. To test for interspecific differences in the daphnids' tolerance to cyanobacteria, their somatic and population growth rates were determined for each D. magna and D. pulex clone after exposure to varying concentrations of two Microcystis aeruginosa strains. The M. aeruginosa strains NIVA and PCC(-) contained either chymotrypsin or trypsin inhibitors, but no microcystins. Mean somatic and population growth rates on a diet with 20% NIVA were significantly more reduced in D. pulex than in D. magna. On a diet with 10% PCC(-), the population growth of D. pulex was significantly more reduced than that of D. magna. This indicates that D. magna is more tolerant to cyanobacteria with protease inhibitors than D. pulex. The reduction of growth rates was possibly caused by an interference of cyanobacterial inhibitors with proteases in the gut of Daphnia, as many other conceivable factors, which might have been able to explain the reduced growth, could be excluded as causal factors. Protease assays revealed that the sensitivities of chymotrypsins and trypsins to cyanobacterial protease inhibitors did not differ between D. magna and D. pulex. However, D. magna exhibited a 2.3-fold higher specific chymotrypsin activity than D. pulex, which explains the observed higher tolerance to cyanobacterial protease inhibitors of D. magna. The present study suggests that D. magna may control the development of cyanobacterial blooms more efficiently than D. pulex due to differences in their tolerance to cyanobacteria with protease inhibitors.

Modeling of Women's 100-M Dash World Record: Wind-Aided or Not?

ERIC Educational Resources Information Center

Hazelrigg, Conner; Waibel, Bryson; Baker, Blane

2015-01-01

On July 16, 1988, Florence Griffith Joyner (FGJ) shattered the women's 100-m dash world record (WR) with a time of 10.49 s, breaking the previous mark by an astonishing 0.27 s. By all accounts FGJ dominated the race that day, securing her place as the premiere female sprinter of that era, and possibly all time. In the aftermath of such an…

75 FR 45076 - Approval and Promulgation of Air Quality Implementation Plans; New York, New Jersey, and...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-02

... the 2007-2009 design value for the NY-NJ-CT PM 2.5 nonattainment area would be 14.0 [micro]g/m\\3\\. VI... the New York-N. New Jersey-Long Island, NY-NJ-CT fine particle (PM 2.5 ) nonattainment area has... determine that the New York-N. New Jersey-Long Island, NY[dash]NJ[dash]CT PM 2.5 nonattainment area...

Wave Evolution in River Mouths and Tidal Inlets

DTIC Science & Technology

2014-06-01

Monterey Bay by a Datawell Buoy (blue) and three collocated WRD buoys (red). Also shown is the f −4 spectral roll off (black dashed). .............. 48...f −4 spectral roll off (black dashed) and the blocking frequency in regions B-E. .................................................... 53   Figure...Significant Wave Height Hz hertz IMU Inertial measurement unit JONSWAP Joint North Sea Wave Program km kilometer MCR Mouth of the Columbia River MEMS

Relationship between self-reported upper limb disability and quantitative tests in hand-arm vibration syndrome.

PubMed

Poole, Kerry; Mason, Howard

2007-03-15

To establish the relationship between quantitative tests of hand function and upper limb disability, as measured by the Disability of the Arm, Shoulder and Hand (DASH) questionnaire, in hand-arm vibration syndrome (HAVS). A total of 228 individuals with HAVS were included in this study. Each had undergone a full HAVS assessment by an experienced physician, including quantitative tests of vibrotactile and thermal perception thresholds, maximal hand-grip strength (HG) and the Purdue pegboard (PP) test. Individuals were also asked to complete a DASH questionnaire. PP and HG of the quantitative tests gave the best and statistically significant individual correlations with the DASH disability score (r2 = 0.168 and 0.096). Stepwise linear regression analysis revealed that only PP and HG measurements were statistically significant predictors of upper limb disability (r2 = 0.178). Overall a combination of the PP and HG measurements, rather than each alone, gave slightly better discrimination, although not statistically significant, between normal and abnormal DASH scores with a sensitivity of 73.1% and specificity of 64.3%. Measurements of manual dexterity and hand-grip strength using PP and HG may be useful in helping to confirm lack of upper limb function and 'perceived' disability in HAVS.

Mediterranean Diet and Other Dietary Patterns in Primary Prevention of Heart Failure and Changes in Cardiac Function Markers: A Systematic Review.

PubMed

Sanches Machado d'Almeida, Karina; Ronchi Spillere, Stefanny; Zuchinali, Priccila; Corrêa Souza, Gabriela

2018-01-10

Heart failure (HF) is a complex syndrome and is recognized as the ultimate pathway of cardiovascular disease (CVD). Studies using nutritional strategies based on dietary patterns have proved to be effective for the prevention and treatment of CVD. Although there are studies that support the protective effect of these diets, their effects on the prevention of HF are not clear yet. We searched the Medline, Embase, and Cochrane databases for studies that examined dietary patterns, such as dietary approaches to stop hypertension (DASH diet), paleolithic, vegetarian, low-carb and low-fat diets and prevention of HF. No limitations were used during the search in the databases. A total of 1119 studies were identified, 14 met the inclusion criteria. Studies regarding the Mediterranean, DASH, vegetarian, and Paleolithic diets were found. The Mediterranean and DASH diets showed a protective effect on the incidence of HF and/or worsening of cardiac function parameters, with a significant difference in relation to patients who did not adhere to these dietary patterns. It is observed that the adoption of Mediterranean or DASH-type dietary patterns may contribute to the prevention of HF, but these results need to be analyzed with caution due to the low quality of evidence.

Mediterranean Diet and Other Dietary Patterns in Primary Prevention of Heart Failure and Changes in Cardiac Function Markers: A Systematic Review

PubMed Central

Sanches Machado d’Almeida, Karina; Ronchi Spillere, Stefanny; Zuchinali, Priccila; Corrêa Souza, Gabriela

2018-01-01

Background: Heart failure (HF) is a complex syndrome and is recognized as the ultimate pathway of cardiovascular disease (CVD). Studies using nutritional strategies based on dietary patterns have proved to be effective for the prevention and treatment of CVD. Although there are studies that support the protective effect of these diets, their effects on the prevention of HF are not clear yet. Methods: We searched the Medline, Embase, and Cochrane databases for studies that examined dietary patterns, such as dietary approaches to stop hypertension (DASH diet), paleolithic, vegetarian, low-carb and low-fat diets and prevention of HF. No limitations were used during the search in the databases. Results: A total of 1119 studies were identified, 14 met the inclusion criteria. Studies regarding the Mediterranean, DASH, vegetarian, and Paleolithic diets were found. The Mediterranean and DASH diets showed a protective effect on the incidence of HF and/or worsening of cardiac function parameters, with a significant difference in relation to patients who did not adhere to these dietary patterns. Conclusions: It is observed that the adoption of Mediterranean or DASH-type dietary patterns may contribute to the prevention of HF, but these results need to be analyzed with caution due to the low quality of evidence. PMID:29320401

A de novo variant in the ASPRV1 gene in a dog with ichthyosis.

PubMed

Bauer, Anina; Waluk, Dominik P; Galichet, Arnaud; Timm, Katrin; Jagannathan, Vidhya; Sayar, Beyza S; Wiener, Dominique J; Dietschi, Elisabeth; Müller, Eliane J; Roosje, Petra; Welle, Monika M; Leeb, Tosso

2017-03-01

Ichthyoses are a heterogeneous group of inherited cornification disorders characterized by generalized dry skin, scaling and/or hyperkeratosis. Ichthyosis vulgaris is the most common form of ichthyosis in humans and caused by genetic variants in the FLG gene encoding filaggrin. Filaggrin is a key player in the formation of the stratum corneum, the uppermost layer of the epidermis and therefore crucial for barrier function. During terminal differentiation of keratinocytes, the precursor profilaggrin is cleaved by several proteases into filaggrin monomers and eventually processed into free amino acids contributing to the hydration of the cornified layer. We studied a German Shepherd dog with a novel form of ichthyosis. Comparing the genome sequence of the affected dog with 288 genomes from genetically diverse non-affected dogs we identified a private heterozygous variant in the ASPRV1 gene encoding "aspartic peptidase, retroviral-like 1", which is also known as skin aspartic protease (SASPase). The variant was absent in both parents and therefore due to a de novo mutation event. It was a missense variant, c.1052T>C, affecting a conserved residue close to an autoprocessing cleavage site, p.(Leu351Pro). ASPRV1 encodes a retroviral-like protease involved in profilaggrin-to-filaggrin processing. By immunofluorescence staining we showed that the filaggrin expression pattern was altered in the affected dog. Thus, our findings provide strong evidence that the identified de novo variant is causative for the ichthyosis in the affected dog and that ASPRV1 plays an essential role in skin barrier formation. ASPRV1 is thus a novel candidate gene for unexplained human forms of ichthyoses.

The influence of parachute-resisted sprinting on running mechanics in collegiate track athletes.

PubMed

Paulson, Sally; Braun, William A

2011-06-01

The influence of parachute-resisted sprinting on running mechanics in collegiate track athletes. The aim of this investigation was to compare the acute effects of parachute-resisted (PR) sprinting on selected kinematic variables. Twelve collegiate sprinters (mean age 19.58 ± 1.44 years, mass 69.32 ± 14.38 kg, height 1.71 ± 9.86 m) ran a 40-yd dash under 2 conditions: PR sprint and sprint without a parachute (NC) that were recorded on a video computer system (60 Hz). Sagittal plane kinematics of the right side of the body was digitized to calculate joint angles at initial ground contact (IGC) and end ground contact (EGC), ground contact (GC) time, stride rate (SR), stride length (SL), and the times of the 40-yd dashes. The NC 40-yd dash time was significantly faster than the PR trial (p < 0.05). The shoulder angle at EGC significantly increased from 34.10 to 42.10° during the PR trial (p < 0.05). There were no significant differences in GC time, SR, SL, or the other joint angles between the 2 trials (p > 0.05). This study suggests that PR sprinting does not acutely affect GC time, SR, SL and upper extremity or lower extremity joint angles during weight acceptance (IGC) in collegiate sprinters. However, PR sprinting increased shoulder flexion by 23.5% at push-off and decreased speed by 4.4%. While sprinting with the parachute, the athlete's movement patterns resembled their mechanics during the unloaded condition. This indicates the external load caused by PR did not substantially overload the runner, and only caused a minor change in the shoulder during push-off. This sports-specific training apparatus may provide coaches with another method for training athletes in a sports-specific manner without causing acute changes to running mechanics.

Nanothermodynamics Applied to Thermal Processes in Heterogeneous Materials

DTIC Science & Technology

2012-08-03

models agree favorably with a wide range of measurements of local thermal and dynamic properties. Progress in understanding basic thermodynamic...Monte- Carlo (MC) simulations of the Ising model .7 The solid black lines in Fig. 4 show results using the uncorrected (Metropolis) algorithm on the...parameter g=0.5 (green, dash-dot), g=1 (black, solid ), and g=2 (blue, dash-dot-dot). Note the failure of the standard Ising model (g=0) to match

User’s Manual for the Modular Analysis-Package Libraries ANAPAC and TRANL

DTIC Science & Technology

1977-09-01

number) Computer software Fourier transforms Computer software library Interpolation software Digitized data...disregarded to give the user a simplified plot. (b) The last digit of ISPACE determines the type of line to be drawn, provided KODE is not...negative. If the last digit of ISPACE is 0 a solid line is drawn 1 a dashed line is drawn - - - 2 a dotted line is drawn .... 3 a dash-dot line is

Transient Simulation of Ram Accelerator Flowfields

DTIC Science & Technology

1993-01-01

PROPULSIVE FLOWS WITH COMIBUSTION CHEMISTRY, ADVANCED TURBULENCE MODELS - 1񕚑ŕ. Reference: Dash, "Advanced Computational Models for Analyzing High - Speed...coupled, implicit manner. Near-wall effects have been dealt with via the low Reynolds number formulation of Chien: and the recent model of Rodi.3 High ...July 1989. 19 Dash, S.M., ’Advanced Computational Models for Analyzing High -Speed Propulsive Flowficlds,’ 199M J&ý,NAF Propulsion Meeting, CPIA Pub. 550

Valve and dash-pot assembly

DOEpatents

Chang, Shih-Chih

1986-01-01

A dash-pot valve comprising a cylinder submerged in the fluid of a housing and having a piston attached to a plunger projecting into the path of closing movement of a pivotal valve member. A vortex chamber in said cylinder is provided with tangentially directed inlets to generate vortex flow upon retraction of said plunger and effect increasing resistance against said piston to progressively retard the closing rate of said valve member toward its seat.

Improved valve and dash-pot assembly

DOEpatents

Chang, S.C.

1985-04-23

A dash-pot valve comprises a cylinder submerged in the fluid of a housing and have a piston attached to a plunger projecting into the path of closing movement of a pivotal valve member. A vortex chamber in said cylinder is provided with targentially directed inlets to generate vortex flow upon retraction of said plunger and effect increasing resistance against said piston to progressively retard the closing rate of said valve member toward its seat.

Modular Electronics for Flash Memory Production

DTIC Science & Technology

2011-12-28

DEFENSE TECHNICAL INFORMATION CENTER ImuM ktkUmlimäj DTICfhas determined on oc öf^H AJI that this Technical Document has the Distribution...the second harmonic (8I2/8V2), and a normalization to remove the scale of the measured current. The red dashed lines represent notable vibrational...superimposed as red dashed lines. The agreement between the two spectroscopies is conclusive that we have successfully put our OPE derivative into the gap

Mission Connect Mild TBI Translational Research Consortium

DTIC Science & Technology

2010-08-01

Grill RJ, Parsley MA, Unabia G, Hulsebosch CE. Anti- inflammatory treatment of traumatic brain injury (TBI). Society for Neuroscience, 2009...Hulsebosch, CD, DeWitt DS, Dash PK, Grill RJ, Parsley MA, Unabia, BC, Rea HC, & Perez-Polo, JR Role of IL-1 and TNF receptor activation in neurological...deficits at TBI. Military health Research Forum, 2009. Hulsebosch, CD, DeWitt DS, Dash PK, Grill RJ, Parsley MA, Unabia, BC, Rea HC, & Perez-Polo, JR

Organic Electronic Devices Using Crosslinked Polyelectrolyte Multilayers as an Ultra-Thin Dielectric Material

DTIC Science & Technology

2006-09-01

energy band diagram illustrating the allowed energies for valence and conducting electrons. The dashes within the band gap (Eg) represent localized ...allowed energies for valence and conducting electrons. The dashes within the band gap (Eg) represent localized electron energy states, or traps, that...been observed with the formation of alternating bond lengths along the backbone.43 The localization of the π-electrons while forming the shorter double

Engaging Students via Innovative Military Useful Technologies

DTIC Science & Technology

2013-07-01

projects involved four additional M.Eng. students who are all finishing their degrees during the spring of 2013: Aniket Dash, Sunil Channapatna...Ravindrachar, Herat Gandhi, and Heesung Sohn. Aniket Dash and Sunil Channapatna Ravindrachar have worked with us on creating a way for unmanaged C++, C...16 and was able to get some simple Isis2 system calls working. In the spring semester, Aniket and Sunil have made very strong progress and are

Single-photon emission of InAs/InP quantum dashes at 1.55 μm and temperatures up to 80 K

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dusanowski, Ł., E-mail: lukasz.dusanowski@pwr.edu.pl; Syperek, M.; Misiewicz, J.

2016-04-18

We report on single photon emission from a self-assembled InAs/InGaAlAs/InP quantum dash emitting at 1.55 μm at the elevated temperatures. The photon auto-correlation histograms of the emission from a charged exciton indicate clear antibunching dips with as-measured g{sup (2)}(0) values significantly below 0.5 recorded at temperatures up to 80 K. It proves that the charged exciton complex in a single quantum dash of the mature InP-based material system can act as a true single photon source up to at least liquid nitrogen temperature. This demonstrates the huge potential of InAs on InP nanostructures as the non-classical light emitters for long-distance fiber-based securemore » communication technologies.« less

Reductive transformation of V(iii) precursors into vanadium(ii) oxide nanowires.

PubMed

Ojelere, Olusola; Graf, David; Ludwig, Tim; Vogt, Nicholas; Klein, Axel; Mathur, Sanjay

2018-05-15

Vanadium(ii) oxide nanostructures are promising materials for supercapacitors and electrocatalysis because of their excellent electrochemical properties and high surface area. In this study, new homoleptic vanadium(iii) complexes with bi-dentate O,N-chelating heteroarylalkenol ligands (DmoxCH[double bond, length as m-dash]COCF3, PyCH[double bond, length as m-dash]COCF3 and PyN[double bond, length as m-dash]COCF3) were synthesized and successfully transformed by reductive conversion into VO nanowires. The chemical identity of V(iii) complexes and their redox behaviour were unambiguously established by single crystal X-ray diffraction studies, cyclic voltammetry, spectrometric studies and DFT calculations. Transformation into the metastable VO phase was verified by powder X-ray diffraction and thermo-gravimetry. Transmission electron microscopy and X-ray photoelectron spectroscopy data confirmed the morphology and chemical composition of VO nanostructures, respectively.

Musculoskeletal system problems and quality of life of mothers of children with cerebral palsy with different levels of disability.

PubMed

Kavlak, Erdoğan; Altuğ, Filiz; Büker, Nihal; Şenol, Hande

2015-01-01

The objective of this study is to investigate musculoskeletal system problems and quality of life of mothers of children with cerebral palsy with different levels of disability. 100 children (37 girls and 63 boys) with cerebral palsy (CP) and their mothers were included in this study. Functional levels of children with CP were assessed by using the Gross Motor Function Classification System (GMFCS) and the Pediatric Functional Independence Measure (WeeFIM). Quality of life of mothers regarding health was assessed by using the Nottingham Health Profile (NHP). Musculoskeletal system problems of mothers were assessed by using the Neck Disability Index (NDI) and the Roland-Morris Disability Questionnaire (RMDQ). No statistical significance was found when GMFCS levels of children with CP and the NHP, DASH-T, RMDQ, NDI and the BAE values of mothers were compared in an inter-group way (p> 0.05). When the NHP parameters and the existence of lower and arm pains of mothers were compared with their BAI, NDI, RMDQ and DASH-T scores, a statistically significant relationship was found among them (p< 0.05). As functional levels of children with CP get worse, upper extremity, lower back and neck problems and anxiety levels of mothers increase and this situation negatively affects mothers' quality of life.

A web-based nutrition program reduces health care costs in employees with cardiac risk factors: before and after cost analysis.

PubMed

Sacks, Naomi; Cabral, Howard; Kazis, Lewis E; Jarrett, Kelli M; Vetter, Delia; Richmond, Russell; Moore, Thomas J

2009-10-23

Rising health insurance premiums represent a rapidly increasing burden on employer-sponsors of health insurance and their employees. Some employers have become proactive in managing health care costs by providing tools to encourage employees to directly manage their health and prevent disease. One example of such a tool is DASH for Health, an Internet-based nutrition and exercise behavior modification program. This program was offered as a free, opt-in benefit to US-based employees of the EMC Corporation. The aim was to determine whether an employer-sponsored, Internet-based diet and exercise program has an effect on health care costs. There were 15,237 total employees and spouses who were included in our analyses, of whom 1967 enrolled in the DASH for Health program (DASH participants). Using a retrospective, quasi-experimental design, study year health care costs among DASH participants and non-participants were compared, controlling for baseline year costs, risk, and demographic variables. The relationship between how often a subject visited the DASH website and health care costs also was examined. These relationships were examined among all study subjects and among a subgroup of 735 subjects with cardiovascular conditions (diabetes, hypertension, hyperlipidemia). Multiple linear regression analysis examined the relationship of program use to health care costs, comparing study year costs among DASH participants and non-participants and then examining the effects of increased website use on health care costs. Analyses were repeated among the cardiovascular condition subgroups. Overall, program use was not associated with changes in health care costs. However, among the cardiovascular risk study subjects, health care costs were US$827 lower, on average, during the study year (P= .05; t(729) = 1.95). Among 1028 program users, increased website use was significantly associated with lower health care costs among those who visited the website at least nine times during the study year (US$14 decrease per visit; P = .04; t(1022) = 2.05), with annual savings highest among 80 program users with targeted conditions (US$55 decrease per visit; P < .001; t(74) = 2.71). An employer-sponsored, Internet-based diet and exercise program shows promise as a low-cost benefit that contributes to lower health care costs among persons at higher risk for above-average health care costs and utilization.

A Web-Based Nutrition Program Reduces Health Care Costs in Employees With Cardiac Risk Factors: Before and After Cost Analysis

PubMed Central

Cabral, Howard; Kazis, Lewis E; Jarrett, Kelli M; Vetter, Delia; Richmond, Russell; Moore, Thomas J

2009-01-01

Background Rising health insurance premiums represent a rapidly increasing burden on employer-sponsors of health insurance and their employees. Some employers have become proactive in managing health care costs by providing tools to encourage employees to directly manage their health and prevent disease. One example of such a tool is DASH for Health, an Internet-based nutrition and exercise behavior modification program. This program was offered as a free, opt-in benefit to US-based employees of the EMC Corporation. Objective The aim was to determine whether an employer-sponsored, Internet-based diet and exercise program has an effect on health care costs. Methods There were 15,237 total employees and spouses who were included in our analyses, of whom 1967 enrolled in the DASH for Health program (DASH participants). Using a retrospective, quasi-experimental design, study year health care costs among DASH participants and non-participants were compared, controlling for baseline year costs, risk, and demographic variables. The relationship between how often a subject visited the DASH website and health care costs also was examined. These relationships were examined among all study subjects and among a subgroup of 735 subjects with cardiovascular conditions (diabetes, hypertension, hyperlipidemia). Multiple linear regression analysis examined the relationship of program use to health care costs, comparing study year costs among DASH participants and non-participants and then examining the effects of increased website use on health care costs. Analyses were repeated among the cardiovascular condition subgroups. Results Overall, program use was not associated with changes in health care costs. However, among the cardiovascular risk study subjects, health care costs were US$827 lower, on average, during the study year (P = .05; t 729 = 1.95). Among 1028 program users, increased website use was significantly associated with lower health care costs among those who visited the website at least nine times during the study year (US$14 decrease per visit; P = .04; t 1022 = 2.05), with annual savings highest among 80 program users with targeted conditions (US$55 decrease per visit; P < .001; t 74 = 2.71). Conclusions An employer-sponsored, Internet-based diet and exercise program shows promise as a low-cost benefit that contributes to lower health care costs among persons at higher risk for above-average health care costs and utilization. PMID:19861297

Neutrophil Elastase, Proteinase 3, and Cathepsin G as Therapeutic Targets in Human Diseases

PubMed Central

Horwitz, Marshall S.; Jenne, Dieter E.; Gauthier, Francis

2010-01-01

Polymorphonuclear neutrophils are the first cells recruited to inflammatory sites and form the earliest line of defense against invading microorganisms. Neutrophil elastase, proteinase 3, and cathepsin G are three hematopoietic serine proteases stored in large quantities in neutrophil cytoplasmic azurophilic granules. They act in combination with reactive oxygen species to help degrade engulfed microorganisms inside phagolysosomes. These proteases are also externalized in an active form during neutrophil activation at inflammatory sites, thus contributing to the regulation of inflammatory and immune responses. As multifunctional proteases, they also play a regulatory role in noninfectious inflammatory diseases. Mutations in the ELA2/ELANE gene, encoding neutrophil elastase, are the cause of human congenital neutropenia. Neutrophil membrane-bound proteinase 3 serves as an autoantigen in Wegener granulomatosis, a systemic autoimmune vasculitis. All three proteases are affected by mutations of the gene (CTSC) encoding dipeptidyl peptidase I, a protease required for activation of their proform before storage in cytoplasmic granules. Mutations of CTSC cause Papillon-Lefèvre syndrome. Because of their roles in host defense and disease, elastase, proteinase 3, and cathepsin G are of interest as potential therapeutic targets. In this review, we describe the physicochemical functions of these proteases, toward a goal of better delineating their role in human diseases and identifying new therapeutic strategies based on the modulation of their bioavailability and activity. We also describe how nonhuman primate experimental models could assist with testing the efficacy of proposed therapeutic strategies. PMID:21079042

Effects of protease and non-starch polysaccharide enzyme on performance, digestive function, activity and gene expression of endogenous enzyme of broilers.

PubMed

Yuan, Lin; Wang, Mingfa; Zhang, Xiaotu; Wang, Zhixiang

2017-01-01

Three hundred one-day-old male broiler chickens (Ross-308) were fed corn-soybean basal diets containing non-starch polysaccharide (NSP) enzyme and different levels of acid protease from 1 to 42 days of age to investigate the effects of exogenous enzymes on growth performance, digestive function, activity of endogenous digestive enzymes in the pancreas and mRNA expression of pancreatic digestive enzymes. For days 1-42, compared to the control chickens, average daily feed intake (ADFI) and average daily gain (ADG) were significantly enhanced by the addition of NSP enzyme in combination with protease supplementation at 40 or 80 mg/kg (p<0.05). Feed-to-gain ratio (FGR) was significantly improved by supplementation with NSP enzymes or NSP enzyme combined with 40 or 80 mg/kg protease compared to the control diet (p<0.05). Apparent digestibility of crude protein (ADCP) was significantly enhanced by the addition of NSP enzyme or NSP enzyme combined with 40 or 80 mg/kg protease (p<0.05). Cholecystokinin (CCK) level in serum was reduced by 31.39% with NSP enzyme combined with protease supplementation at 160 mg/kg (p<0.05), but the CCK level in serum was increased by 26.51% with NSP enzyme supplementation alone. After 21 days, supplementation with NSP enzyme and NSP enzyme combined with 40 or 80 mg/kg protease increased the activity of pancreatic trypsin by 74.13%, 70.66% and 42.59% (p<0.05), respectively. After 42 days, supplementation with NSP enzyme and NSP enzyme combined with 40 mg/kg protease increased the activity of pancreatic trypsin by 32.45% and 27.41%, respectively (p<0.05). However, supplementation with NSP enzyme and 80 or 160 mg/kg protease decreased the activity of pancreatic trypsin by 10.75% and 25.88%, respectively (p<0.05). The activities of pancreatic lipase and amylase were significantly higher in treated animals than they were in the control group (p<0.05). Supplementation with NSP enzyme, NSP enzyme combined with 40 or 80 mg/kg protease increased pancreatic trypsin mRNA levels by 40%, 44% and 28%, respectively. Supplementation with NSP enzyme and 160 mg/kg protease decreased pancreatic trypsin mRNA levels by 13%. Pancreatic lipase and amylase mRNA expression were significantly elevated in treated animals compared to the control group (p<0.05). These results suggest that the amount of NSP enzyme and acid protease in the diet significantly affects digestive function, endogenous digestive-enzyme activity and mRNA expression in broilers.

Human eosinophils constitutively express a unique serine protease, PRSS33.

PubMed

Toyama, Sumika; Okada, Naoko; Matsuda, Akio; Morita, Hideaki; Saito, Hirohisa; Fujisawa, Takao; Nakae, Susumu; Karasuyama, Hajime; Matsumoto, Kenji

2017-07-01

Eosinophils play important roles in asthma, especially airway remodeling, by producing various granule proteins, chemical mediators, cytokines, chemokines and proteases. However, protease production by eosinophils is not fully understood. In the present study, we investigated the production of eosinophil-specific proteases/proteinases by transcriptome analysis. Human eosinophils and other cells were purified from peripheral blood by density gradient sedimentation and negative/positive selections using immunomagnetic beads. Protease/proteinase expression in eosinophils and release into the supernatant were evaluated by microarray analysis, qPCR, ELISA, flow cytometry and immunofluorescence staining before and after stimulation with eosinophil-activating cytokines and secretagogues. mRNAs for extracellular matrix proteins in human normal fibroblasts were measured by qPCR after exposure to recombinant protease serine 33 (PRSS33) protein (rPRSS33), created with a baculovirus system. Human eosinophils expressed relatively high levels of mRNA for metalloproteinase 25 (MMP25), a disintegrin and metalloprotease 8 (ADAM8), ADAM10, ADAM19 and PRSS33. Expression of PRSS33 was the highest and eosinophil-specific. PRSS33 mRNA expression was not affected by eosinophil-activating cytokines. Immunofluorescence staining showed that PRSS33 was co-localized with an eosinophil granule protein. PRSS33 was not detected in the culture supernatant of eosinophils even after stimulation with secretagogues, but its cell surface expression was increased. rPRSS33 stimulation of human fibroblasts increased expression of collagen and fibronectin mRNAs, at least in part via protease-activated receptor-2 activation. Activated eosinophils may induce fibroblast extracellular matrix protein synthesis via cell surface expression of PRSS33, which would at least partly explain eosinophils' role(s) in airway remodeling. Copyright © 2017 Japanese Society of Allergology. Production and hosting by Elsevier B.V. All rights reserved.

Accelerated Neuronal Cell Recovery from Botulinum Neurotoxin Intoxication by Targeted Ubiquitination

PubMed Central

Kuo, Chueh-Ling; Oyler, George A.; Shoemaker, Charles B.

2011-01-01

Botulinum neurotoxin (BoNT), a Category A biodefense agent, delivers a protease to motor neuron cytosol that cleaves one or more soluble NSF attachment protein receptors (SNARE) proteins involved in neurotransmission to cause a flaccid paralysis. No antidotes exist to reverse symptoms of BoNT intoxication so severely affected patients require artificial respiration with prolonged intensive care. Time to recovery depends on toxin serotype because the intraneuronal persistence of the seven known BoNT serotypes varies widely from days to many months. Our therapeutic antidote strategy is to develop ‘targeted F-box’ (TFB) agents that target the different intraneuronal BoNT proteases for accelerated degradation by the ubiquitin proteasome system (UPS), thus promoting rapid recovery from all serotypes. These agents consist of a camelid heavy chain-only VH (VHH) domain specific for a BoNT protease fused to an F-box domain recognized by an intraneuronal E3-ligase. A fusion protein containing the 14 kDa anti-BoNT/A protease VHH, ALcB8, joined to a 15 kDa F-box domain region of TrCP (D5) was sufficient to cause increased ubiquitination and accelerate turnover of the targeted BoNT/A protease within neurons. Neuronal cells expressing this TFB, called D5-B8, were also substantially resistant to BoNT/A intoxication and recovered from intoxication at least 2.5 fold quicker than control neurons. Fusion of D5 to a VHH specific for BoNT/B protease (BLcB10) led to accelerated turnover of the targeted protease within neurons, thus demonstrating the modular nature of these therapeutic agents and suggesting that development of similar therapeutic agents specific to all botulinum serotypes should be readily achievable. PMID:21629663

Accelerated neuronal cell recovery from Botulinum neurotoxin intoxication by targeted ubiquitination.

PubMed

Kuo, Chueh-Ling; Oyler, George A; Shoemaker, Charles B

2011-01-01

Botulinum neurotoxin (BoNT), a Category A biodefense agent, delivers a protease to motor neuron cytosol that cleaves one or more soluble NSF attachment protein receptors (SNARE) proteins involved in neurotransmission to cause a flaccid paralysis. No antidotes exist to reverse symptoms of BoNT intoxication so severely affected patients require artificial respiration with prolonged intensive care. Time to recovery depends on toxin serotype because the intraneuronal persistence of the seven known BoNT serotypes varies widely from days to many months. Our therapeutic antidote strategy is to develop 'targeted F-box' (TFB) agents that target the different intraneuronal BoNT proteases for accelerated degradation by the ubiquitin proteasome system (UPS), thus promoting rapid recovery from all serotypes. These agents consist of a camelid heavy chain-only V(H) (VHH) domain specific for a BoNT protease fused to an F-box domain recognized by an intraneuronal E3-ligase. A fusion protein containing the 14 kDa anti-BoNT/A protease VHH, ALcB8, joined to a 15 kDa F-box domain region of TrCP (D5) was sufficient to cause increased ubiquitination and accelerate turnover of the targeted BoNT/A protease within neurons. Neuronal cells expressing this TFB, called D5-B8, were also substantially resistant to BoNT/A intoxication and recovered from intoxication at least 2.5 fold quicker than control neurons. Fusion of D5 to a VHH specific for BoNT/B protease (BLcB10) led to accelerated turnover of the targeted protease within neurons, thus demonstrating the modular nature of these therapeutic agents and suggesting that development of similar therapeutic agents specific to all botulinum serotypes should be readily achievable.

DASH Eating Plan: An Eating Pattern for Diabetes Management

PubMed Central

2017-01-01

IN BRIEF The DASH (Dietary Approaches to Stop Hypertension) eating plan is an acceptable eating pattern for people who have diabetes. In addition to promoting blood pressure control, this eating pattern has been shown to improve insulin resistance, hyperlipidemia, and even overweight/obesity. This balanced approach promotes consumption of a variety of foods (whole grains, fat-free or low-fat dairy products, fruits, vegetables, poultry, fish, and nuts) and is appropriate for the entire family. PMID:28588372

Combat Casualty Hand Burns: Evaluating Impairment and Disability during Recovery

DTIC Science & Technology

2008-06-01

impairment guidelines would correlate with disability as mea- sured by the DASH. However, a study by Mink van der Molen et al. found only a weak correlation (r...Mink van der Molen AB, Ettema AM, Hovius SER. Outcome of hand trauma: the hand injury severity scoring system (HISS) and subsequent impairment and...0.38) between AMA and DASH scores at six months after hand trauma.16 In another study, van Oosterom et al. reported no statistically significant

Mission Connect Mild TBI Translational Research Consortium

DTIC Science & Technology

2009-08-01

modalities. We recruited to the project Margaret A. Parsley to carry out the Mild Traumatic Brain Injuries (MTBIs) under the supervision of Doug DeWitt...abstracts: Hulsebosch CE, Johnson KM, Dewitt DS, Dash PK, Grill R, Parsley MA, Unabia G, Rea H, Perez-Polo JR. Role of IL-1 and TNF receptor activation in...neurological deficits after TBI National Neurotrauma Society, 2010 Johnson KM, Dewitt DS, Dash PK, Grill R, Parsley MA, Unabia G, Rea H, Perez-Polo

Dynamic Testing of Laterally Confined Concrete

DTIC Science & Technology

1990-09-01

for Intermediate Confining pressure (Dashed Curve). 31 23. Example of Regression Fit by Equation (6) for Highest Pressure Group (Dashed Curve... pressure group , loaded by a moderate striker-bar impact speed of 420 in/sec. (10.7 m/s). The peak stress of 124 MPa (18 ksi) occurs at a strain of...survived at one end. This was for the highest speed impact in the lowest confining pressure group . Curves are given in the Appendix Figure A-15. The

Ammonolysis of ketene as a potential source of acetamide in the troposphere: a quantum chemical investigation.

PubMed

Sarkar, Saptarshi; Mallick, Subhasish; Kumar, Pradeep; Bandyopadhyay, Biman

2018-05-16

Quantum chemical calculations at the CCSD(T)/CBS//MP2/aug-cc-pVTZ levels of theory have been carried out to investigate a potential new source of acetamide in Earth's atmosphere through the ammonolysis of the simplest ketene. It was found that the reaction can occur via the addition of ammonia at either the C[double bond, length as m-dash]C or C[double bond, length as m-dash]O bond of ketene. The potential energy surface as well as calculated rate coefficients indicate that under tropospheric conditions, ammonolysis would occur almost exclusively via ammonia addition at the C[double bond, length as m-dash]O bond with negligible contribution from addition at the C[double bond, length as m-dash]C bond. The reaction of ketene with water has also been investigated in order to compare between hydrolysis and ammonolysis, as the former is known to be responsible for the formation of acetic acid. The rate coefficient for the formation of acetamide was found to be ∼106 to 109 times higher than that for the formation of acetic acid from the same ketene source in the troposphere. By means of the relative rate of ammonolysis with respect to hydrolysis, it was shown that acetamide formation would dominate over acetic acid formation at various altitudes in the troposphere.

Pressure-induced emission band separation of the hybridized local and charge transfer excited state in a TPE-based crystal.

PubMed

Liu, Xuedan; Li, Aisen; Xu, Weiqing; Ma, Zhiyong; Jia, Xinru

2018-05-08

We herein report a newly synthesized simple molecule, named TPE[double bond, length as m-dash]C4, with twisted D-A structure. TPE[double bond, length as m-dash]C4 showed two intrinsic emission bands ascribed to the locally excited (LE) state and the intramolecular charge transfer (ICT) state, respectively. In the crystal state, the LE emission band is usually observed. However, by applying hydrostatic pressure to the powder sample and the single crystal sample of TPE[double bond, length as m-dash]C4, dual-fluorescence (445 nm and 532 nm) was emerged under high pressure, owing to the pressure-induced emission band separation of the hybridized local and charge transfer excited state (HLCT). It is found that the emission of TPE[double bond, length as m-dash]C4 is generally determined by the ratio of the LE state to the ICT state. The ICT emission band is much more sensitive to the external pressure than the LE emission band. The HLCT state leads to a sample with different responsiveness to grinding and hydrostatic pressure. This study is of significance in the molecular design of such D-A type molecules and in the control of photoluminescence features by molecular structure. Such results are expected to pave a new way to further understand the relationship between the D-A molecular structure and stimuli-responsive properties.

Pilot randomised trial of a healthy eating behavioural intervention in uncontrolled asthma.

PubMed

Ma, Jun; Strub, Peg; Lv, Nan; Xiao, Lan; Camargo, Carlos A; Buist, A Sonia; Lavori, Philip W; Wilson, Sandra R; Nadeau, Kari C; Rosas, Lisa G

2016-01-01

Rigorous research on the benefit of healthy eating patterns for asthma control is lacking.We randomised 90 adults with objectively confirmed uncontrolled asthma and a low-quality diet (Dietary Approaches to Stop Hypertension (DASH) scores <6 out of 9) to a 6-month DASH behavioural intervention (n=46) or usual-care control (n=44). Intention-to-treat analyses used repeated-measures mixed models.Participants were middle-aged, 67% female and multiethnic. Compared with controls, intervention participants improved on DASH scores (mean change (95% CI) 0.6 (0, 1.1) versus -0.3 (-0.8, 0.2); difference 0.8 (0.2, 1.5)) and the primary outcome, Asthma Control Questionnaire scores (-0.2 (-0.5, 0) versus 0 (-0.3, 0.3); difference -0.2 (-0.5, 0.1)) at 6 months. The mean group differences in changes in Mini Asthma Quality of Life Questionnaire overall and subdomain scores consistently favoured the intervention over the control group: overall 0.4 (95% CI 0, 0.8), symptoms 0.5 (0, 0.9), environment 0.4 (-0.1, 1.0), emotions 0.4 (-0.2, 0.9) and activities 0.3 (0, 0.7). These differences were modest, but potentially clinical significant.The DASH behavioural intervention improved diet quality with promising clinical benefits for better asthma control and functional status among adults with uncontrolled asthma. A full-scale efficacy trial is warranted. Copyright ©ERS 2016.

Pre-equilibrium competitive library screening for tuning inhibitor association rate and specificity toward serine proteases.

PubMed

Cohen, Itay; Naftaly, Si; Ben-Zeev, Efrat; Hockla, Alexandra; Radisky, Evette S; Papo, Niv

2018-04-16

High structural and sequence similarity within protein families can pose significant challenges to the development of selective inhibitors, especially toward proteolytic enzymes. Such enzymes usually belong to large families of closely similar proteases and may also hydrolyze, with different rates, protein- or peptide-based inhibitors. To address this challenge, we employed a combinatorial yeast surface display library approach complemented with a novel pre-equilibrium, competitive screening strategy for facile assessment of the effects of multiple mutations on inhibitor association rates and binding specificity. As a proof of principle for this combined approach, we utilized this strategy to alter inhibitor/protease association rates and to tailor the selectivity of the amyloid β-protein precursor Kunitz protease inhibitor domain (APPI) for inhibition of the oncogenic protease mesotrypsin, in the presence of three competing serine proteases, anionic trypsin, cationic trypsin and kallikrein-6. We generated a variant, designated APPI P13W/M17G/I18F/F34V , with up to 30-fold greater specificity relative to the parental APPI M17G/I18F/F34V protein, and 6500- to 230 000-fold improved specificity relative to the wild-type APPI protein in the presence of the other proteases tested. A series of molecular docking simulations suggested a mechanism of interaction that supported the biochemical results. These simulations predicted that the selectivity and specificity are affected by the interaction of the mutated APPI residues with nonconserved enzyme residues located in or near the binding site. Our strategy will facilitate a better understanding of the binding landscape of multispecific proteins and will pave the way for design of new drugs and diagnostic tools targeting proteases and other proteins. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Molecular principles behind Boceprevir resistance due to mutations in hepatitis C NS3/4A protease.

PubMed

Nagpal, Neha; Goyal, Sukriti; Wahi, Divya; Jain, Ritu; Jamal, Salma; Singh, Aditi; Rana, Preeti; Grover, Abhinav

2015-10-01

The hepatitis C virus (HCV) infection is a primary cause of chronic hepatitis which eventually progresses to cirrhosis and in some instances might advance to hepatocellular carcinoma. According to the WHO report, HCV infects 130-150 million people globally and every year 350,000 to 500,000 people die from hepatitis C virus infection. Great achievement has been made in viral treatment evolution, after the development of HCV NS3/4A protease inhibitor (Boceprevir). However, efficacy of Boceprevir is compromised by the emergence of drug resistant variants. The molecular principle behind drug resistance of the protease mutants such as (V36M, T54S and R155K) is still poorly understood. Therefore in this study, we employed a series of computational strategies to analyze the binding of antiviral drug, Boceprevir to HCV NS3/4A protease mutants. Our results clearly demonstrate that the point mutations (V36M, T54S and R155K) in protease are associated with lowering of its binding affinity with Boceprevir. Exhaustive analysis of the simulated Boceprevir-bound wild and mutant complexes revealed variations in hydrophobic interactions, hydrogen bond occupancy and salt bridge interactions. Also, substrate envelope analysis scrutinized that the studied mutations reside outside the substrate envelope which may affect the Boceprevir affinity towards HCV protease but not the protease enzymatic activity. Furthermore, structural analyses of the binding site volume and flexibility show impairment in flexibility and stability of the binding site residues in mutant structures. In order to combat Boceprevir resistance, renovation of binding interactions between the drug and protease may be valuable. The structural insight from this study reveals the mechanism of the Boceprevir resistance and the results can be valuable for the design of new PIs with improved efficiency. Copyright © 2015 Elsevier B.V. All rights reserved.

Biochemical characterization of recombinant Enterovirus 71 3C protease with fluorogenic model peptide substrates and development of a biochemical assay.

PubMed

Shang, Luqing; Zhang, Shumei; Yang, Xi; Sun, Jixue; Li, Linfeng; Cui, Zhengjie; He, Qiuhong; Guo, Yu; Sun, Yuna; Yin, Zheng

2015-04-01

Enterovirus 71 (EV71), a primary pathogen of hand, foot, and mouth disease (HFMD), affects primarily infants and children. Currently, there are no effective drugs against HFMD. EV71 3C protease performs multiple tasks in the viral replication, which makes it an ideal antiviral target. We synthesized a small set of fluorogenic model peptides derived from cleavage sites of EV71 polyprotein and examined their efficiencies of cleavage by EV71 3C protease. The novel peptide P08 [(2-(N-methylamino)benzoyl) (NMA)-IEALFQGPPK(DNP)FR] was determined to be the most efficiently cleaved by EV71 3C protease, with a kinetic constant kcat/Km of 11.8 ± 0.82 mM(-1) min(-1). Compared with literature reports, P08 gave significant improvement in the signal/background ratio, which makes it an attractive substrate for assay development. A Molecular dynamics simulation study elaborated the interactions between substrate P08 and EV71 3C protease. Arg39, which is located at the bottom of the S2 pocket of EV71 3C protease, may participate in the proteolysis process of substrates. With an aim to evaluate EV71 3C protease inhibitors, a reliable and robust biochemical assay with a Z' factor of 0.87 ± 0.05 was developed. A novel compound (compound 3) (50% inhibitory concentration [IC50] = 1.89 ± 0.25 μM) was discovered using this assay, which effectively suppressed the proliferation of EV 71 (strain Fuyang) in rhabdomyosarcoma (RD) cells with a highly selective index (50% effective concentration [EC50] = 4.54 ± 0.51 μM; 50% cytotoxic concentration [CC50] > 100 μM). This fast and efficient assay for lead discovery and optimization provides an ideal platform for anti-EV71 drug development targeting 3C protease. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Molecular mechanism of mast cell–mediated innate defense against endothelin and snake venom sarafotoxin

PubMed Central

Schneider, Lars A.; Schlenner, Susan M.; Feyerabend, Thorsten B.; Wunderlin, Markus; Rodewald, Hans-Reimer

2007-01-01

Mast cells are protective against snake venom sarafotoxins that belong to the endothelin (ET) peptide family. The molecular mechanism underlying this recently recognized innate defense pathway is unknown, but secretory granule proteases have been invoked. To specifically disrupt a single protease function without affecting expression of other proteases, we have generated a mouse mutant selectively lacking mast cell carboxypeptidase A (Mc-cpa) activity. Using this mutant, we have now identified Mc-cpa as the essential protective mast cell enzyme. Mass spectrometry of peptide substrates after cleavage by normal or mutant mast cells showed that removal of a single amino acid, the C-terminal tryptophan, from ET and sarafotoxin by Mc-cpa is the principle molecular mechanism underlying this very rapid mast cell response. Mast cell proteases can also cleave ET and sarafotoxin internally, but such “nicking” is not protective because intramolecular disulfide bridges maintain peptide function. We conclude that mast cells attack ET and sarafotoxin exactly at the structure required for toxicity, and hence sarafotoxins could not “evade” Mc-cpa's substrate specificity without loss of toxicity. PMID:17923505

HCV NS3 protease enhances liver fibrosis via binding to and activating TGF-β type I receptor

NASA Astrophysics Data System (ADS)

Sakata, Kotaro; Hara, Mitsuko; Terada, Takaho; Watanabe, Noriyuki; Takaya, Daisuke; Yaguchi, So-Ichi; Matsumoto, Takehisa; Matsuura, Tomokazu; Shirouzu, Mikako; Yokoyama, Shigeyuki; Yamaguchi, Tokio; Miyazawa, Keiji; Aizaki, Hideki; Suzuki, Tetsuro; Wakita, Takaji; Imoto, Masaya; Kojima, Soichi

2013-11-01

Viruses sometimes mimic host proteins and hijack the host cell machinery. Hepatitis C virus (HCV) causes liver fibrosis, a process largely mediated by the overexpression of transforming growth factor (TGF)-β and collagen, although the precise underlying mechanism is unknown. Here, we report that HCV non-structural protein 3 (NS3) protease affects the antigenicity and bioactivity of TGF-β2 in (CAGA)9-Luc CCL64 cells and in human hepatic cell lines via binding to TGF-β type I receptor (TβRI). Tumor necrosis factor (TNF)-α facilitates this mechanism by increasing the colocalization of TβRI with NS3 protease on the surface of HCV-infected cells. An anti-NS3 antibody against computationally predicted binding sites for TβRI blocked the TGF-β mimetic activities of NS3 in vitro and attenuated liver fibrosis in HCV-infected chimeric mice. These data suggest that HCV NS3 protease mimics TGF-β2 and functions, at least in part, via directly binding to and activating TβRI, thereby enhancing liver fibrosis.

Nucleation-dependant chemical bonding paradigm: the effect of rare earth ions on the nucleation of urea in aqueous solution.

PubMed

Chen, Xiaoyan; Sun, Congting; Wu, Sixin; Xue, Dongfeng

2017-03-29

Rare earth ions can be used to construct a variety of novel structures and are favorable to chemical bonding regulation and design. In this study, the chemical bonding paradigm between rare earth ions (Ln 3+ ) and urea molecules in an aqueous solution can be tracked by the evolution of C[double bond, length as m-dash]O, NH 2 , and CN vibration bands during the urea nucleation stage. Rare earth ions such as La 3+ , Gd 3+ , and Lu 3+ can manipulate the nucleation time of urea via regulating the nucleation-dependant N-C[double bond, length as m-dash]OH-N hydrogen-bonding between urea molecules. Two types of chemical bondings between Ln 3+ and urea molecules have been confirmed, which are Ln 3+ O[double bond, length as m-dash]C-N and Ln 3+ NH 2 -C. Compared with Ln 3+ NH 2 -C, Ln 3+ prefers to coordinate with the O[double bond, length as m-dash]C bond in urea. With a higher concentration of rare earth ions in the solution, some N-C[double bond, length as m-dash]OH-N hydrogen bonds are broken as a consequence of the incorporation of Ln 3+ into the lattice, resulting in the decreased symmetry of local urea molecules in the crystalline nuclei and the consequent Ln 3+ concentration-dependent nucleation time of urea. Moreover, using the ionic electronegativity scale of Ln 3+ , the different effects of La 3+ , Gd 3+ , and Lu 3+ on urea nucleation can be further distinguished. The present study provides basic data for unrevealing the chemical bonding regulation role of rare earth ions in the formation of hydrogen bonded materials, which may give insight into the design and fabrication of novel materials utilizing rare earth ions to adjust the chemical bonding process.

Low-Intensity Sprint Training With Blood Flow Restriction Improves 100-m Dash.

PubMed

Behringer, Michael; Behlau, Daniel; Montag, Johannes C K; McCourt, Molly L; Mester, Joachim

2017-09-01

Behringer, M, Behlau, D, Montag, JCK, McCourt, ML, and Mester, J. Low-intensity sprint training with blood flow restriction improves 100-m dash. J Strength Cond Res 31(9): 2462-2472, 2017-We investigated the effects of practical blood flow restriction (pBFR) of leg muscles during sprint training on the 100-m dash time in well-trained sport students. Participants performed 6 × 100-m sprints at 60-70% of their maximal 100-m sprinting speed twice a week for 6 weeks, either with (intervention group [IG]; n = 12) or without pBFR (control group [CG]; n = 12). The 100-m dash time significantly decreased more in the IG (-0.38 ± 0.24 seconds) than in the CG (-0.16 ± 0.17 seconds). The muscle thickness of the rectus femoris increased only in the IG, whereas no group-by-time interactions were found for the muscle thickness of the biceps femoris and the biceps brachii. The maximal isometric force, measured using a leg press, did not change in either group. However, the rate of force development improved in the IG. Growth hormone, testosterone, insulin-like growth factor 1, and cortisol concentrations did not significantly differ between both groups at any measurement time point (pre, 1 minute, 20 minutes, 120 minutes, and 24 hours after the 6 all-out sprints of the first training session). The muscle damage marker h-FABP increased significantly more in the CG than in the IG. The pBFR improved the 100-m dash time significantly more than low-intensity sprint interval training alone. Other noted benefits of training with pBFR were a decreased level of muscle damage, a greater increase of the rectus femoris muscle thickness, and a higher rate of force development. However, the tested hormones were unable to explain the additional beneficial effects.

Prospective study of Dietary Approaches to Stop Hypertension- and Mediterranean-style dietary patterns and age-related cognitive change: the Cache County Study on Memory, Health and Aging.

PubMed

Wengreen, Heidi; Munger, Ronald G; Cutler, Adele; Quach, Anna; Bowles, Austin; Corcoran, Christopher; Tschanz, Joann T; Norton, Maria C; Welsh-Bohmer, Kathleen A

2013-11-01

Healthy dietary patterns may protect against age-related cognitive decline, but results of studies have been inconsistent. We examined associations between Dietary Approaches to Stop Hypertension (DASH)- and Mediterranean-style dietary patterns and age-related cognitive change in a prospective, population-based study. Participants included 3831 men and women ≥65 y of age who were residents of Cache County, UT, in 1995. Cognitive function was assessed by using the Modified Mini-Mental State Examination (3MS) ≤4 times over 11 y. Diet-adherence scores were computed by summing across the energy-adjusted rank-order of individual food and nutrient components and categorizing participants into quintiles of the distribution of the diet accordance score. Mixed-effects repeated-measures models were used to examine 3MS scores over time across increasing quintiles of dietary accordance scores and individual food components that comprised each score. The range of rank-order DASH and Mediterranean diet scores was 1661-25,596 and 2407-26,947, respectively. Higher DASH and Mediterranean diet scores were associated with higher average 3MS scores. People in quintile 5 of DASH averaged 0.97 points higher than those in quintile 1 (P = 0.001). The corresponding difference for Mediterranean quintiles was 0.94 (P = 0.001). These differences were consistent over 11 y. Higher intakes of whole grains and nuts and legumes were also associated with higher average 3MS scores [mean quintile 5 compared with 1 differences: 1.19 (P < 0.001), 1.22 (P < 0.001), respectively]. Higher levels of accordance with both the DASH and Mediterranean dietary patterns were associated with consistently higher levels of cognitive function in elderly men and women over an 11-y period. Whole grains and nuts and legumes were positively associated with higher cognitive functions and may be core neuroprotective foods common to various healthy plant-centered diets around the globe.

An insight into the sialotranscriptome of the seed-feeding bug, Oncopeltus fasciatus.

PubMed

Francischetti, Ivo M B; Lopes, Angela H; Dias, Felipe A; Pham, Van M; Ribeiro, José M C

2007-09-01

The salivary transcriptome of the seed-feeding hemipteran, Oncopeltus fasciatus (milkweed bug), is described following assembly of 1025 expressed sequence tags (ESTs) into 305 clusters of related sequences. Inspection of these sequences reveals abundance of low complexity, putative secreted products rich in the amino acids (aa) glycine, serine or threonine, which might function as silk or mucins and assist food canal lubrication and sealing of the feeding site around the mouthparts. Several protease inhibitors were found, including abundant expression of cystatin transcripts that may inhibit cysteine proteases common in seeds that might injure the insect or induce plant apoptosis. Serine proteases and lipases are described that might assist digestion and liquefaction of seed proteins and oils. Finally, several novel putative proteins are described with no known function that might affect plant physiology or act as antimicrobials.

A Quantitative Analysis of Solar Flare Characteristics as Observed in the Solar Observing Optical Network and the Global Oscillation Network Group

DTIC Science & Technology

2012-03-01

due to their possible threat to operations. Because of their importance, event-level flares are given extra consideration in this report. (AFWA...In the impulsive phase (3 to 10 minutes), as this reconnection takes place, electrons and ions with large energies are accelerated and released...right, correspond to the horizontal lines with large red dashes. The flash and gradual phases are bound by the vertical dashed lines. 13 variable in

Enhancing Sparsity by Reweighted l(1) Minimization

DTIC Science & Technology

2008-07-01

recovery depends on the sparsity level k. The dashed curves represent a reweighted ℓ1 algorithm that outperforms the traditional unweighted ℓ1...approach (solid curve ). (a) Performance after 4 reweighting iterations as a function of ǫ. (b) Performance with fixed ǫ = 0.1 as a function of the number of...signal recovery (declared when ‖x0 − x‖ℓ∞ ≤ 10−3) for the unweighted ℓ1 algorithm as a function of the sparsity level k. The dashed curves represent the

Synthesis and SMM behaviour of trinuclear versus dinuclear 3d-5f uranyl(v)-cobalt(ii) cation-cation complexes.

PubMed

Chatelain, Lucile; Tuna, Floriana; Pécaut, Jacques; Mazzanti, Marinella

2017-05-02

Trinuclear versus dinuclear heterodimetallic U V O 2 + Co 2+ complexes were selectively assembled via a cation-cation interaction by tuning the ligand. The trimeric complex 2, with a linear [Co-O[double bond, length as m-dash]U[double bond, length as m-dash]O-Co] core, exhibits magnetic exchange and slow relaxation with a reversal barrier of 30.5 ± 0.9 K providing the first example of a U-Co exchange-coupled SMM.

47 CFR 15.122 - Closed caption decoder requirements for digital television receivers and converter boxes.

Code of Federal Regulations, 2011 CFR

2011-10-01

... code 0×2D Lower-left border (⌊), G2 char code 0×7C G0 dash (-), char code 0×2D Horizontal border... (-), char code 0×2D Upper-left border (⌈), G2 char code 0×7F G0 dash (-), char code 0×2D (4) Support for..., if the character to the left of the embedded return is a hyphen, the embedded return should be...

47 CFR 15.122 - Closed caption decoder requirements for digital television receivers and converter boxes.

Code of Federal Regulations, 2010 CFR

2010-10-01

... code 0×2D Lower-left border (⌊), G2 char code 0×7C G0 dash (-), char code 0×2D Horizontal border... (-), char code 0×2D Upper-left border (⌈), G2 char code 0×7F G0 dash (-), char code 0×2D (4) Support for..., if the character to the left of the embedded return is a hyphen, the embedded return should be...

Reliability of the detailed assessment of speed of handwriting on Flemish children.

PubMed

Simons, Johan; Probst, Michel

2014-01-01

This study evaluates the reliability of the Detailed Assessment of Speed of Handwriting (DASH) in a Dutch-speaking sample of children. The sample included 650 boys and 513 girls (age range = 9-16 years). Handwriting speed measurements were obtained using the DASH. Interrater agreement, test-retest reliability, and internal consistency were calculated; gender and age effects were analyzed. Interrater agreement shows excellent reliability with intraclass correlation coefficients of at least 0.94. Test-retest correlations ranged from r = 0.65 to r = 0.81. The internal consistency measures, calculated with Cronbach's alpha, were between 0.88 and 0.94. Both gender and age have a significant effect on handwriting speed, with F (7.1144) = 17.43 (P < .001) for gender and F (7.1144) = 21.8 (P < .001) for age. The DASH is a reliable assessment tool to evaluate handwriting speed of Dutch-speaking children. There is a tendency of girls to write faster than boys.

Dots and dashes: art, virtual reality, and the telegraph

NASA Astrophysics Data System (ADS)

Ruzanka, Silvia; Chang, Ben

2009-02-01

Dots and Dashes is a virtual reality artwork that explores online romance over the telegraph, based on Ella Cheever Thayer's novel Wired Love - a Romance in Dots and Dashes (an Old Story Told in a New Way)1. The uncanny similarities between this story and the world of today's virtual environments provides the springboard for an exploration of a wealth of anxieties and dreams, including the construction of identities in an electronically mediated environment, the shifting boundaries between the natural and machine worlds, and the spiritual dimensions of science and technology. In this paper we examine the parallels between the telegraph networks and our current conceptions of cyberspace, as well as unique social and cultural impacts specific to the telegraph. These include the new opportunities and roles available to women in the telegraph industry and the connection between the telegraph and the Spiritualist movement. We discuss the development of the artwork, its structure and aesthetics, and the technical development of the work.

Characteristics of stereotypic movement disorder and self-injurious behavior assessed with the Diagnostic Assessment for the Severely Handicapped (DASH-II).

PubMed

Matson, J L; Hamilton, M; Duncan, D; Bamburg, J; Smiroldo, B; Anderson, S; Baglio, C

1997-01-01

The first experiment involved 143 individuals with severe and profound mental retardation. Individuals with Stereotypic Movement Disorder, Self-Injurious Behavior (SIB), and Stereotypic movement disorder with self-injurious behavior as assessed by the Diagnostic Assessment for the Severely Handicapped-II DASH-II were validated against Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV; American Psychiatric Association, 1994) criteria. In a second study DASH-II scores for 1480 individuals with severe and profound mental retardation were compared on demographic variables, core and associated features of each disorder. Characteristics of persons in each group were reviewed. Persons with profound mental retardation were more likely to evince stereotypies or self-injury compared to their severely impaired counterparts. Also, those with stereotypies were more likely to present with Pervasive Developmental Disorder (PDD)/autism, organicity, and eating disorders, while persons with SIB were more likely to evince sleep, sexual, and eating disorders.

Variably Protease-Sensitive Prionopathy: A New Sporadic Disease of the Prion Protein

PubMed Central

Zou, Wen-Quan; Puoti, Gianfranco; Xiao, Xiangzhu; Yuan, Jue; Qing, Liuting; Cali, Ignazio; Shimoji, Miyuki; Langeveld, Jan P. M.; Castellani, Rudy; Notari, Silvio; Crain, Barbara; Schmidt, Robert E.; Geschwind, Michael; DeArmond, Stephen J.; Cairns, Nigel J.; Dickson, Dennis; Honig, Lawrence; Torres, Juan Maria; Mastrianni, James; Capellari, Sabina; Giaccone, Giorgio; Belay, Ermias D.; Schonberger, Lawrence B.; Cohen, Mark; Perry, George; Kong, Qingzhong; Parchi, Piero; Tagliavini, Fabrizio; Gambetti, Pierluigi

2011-01-01

Objective The objective of the study is to report 2 new genotypic forms of protease-sensitive prionopathy (PSPr), a novel prion disease described in 2008, in 11 subjects all homozygous for valine at codon 129 of the prion protein (PrP) gene (129VV). The 2 new PSPr forms affect individuals who are either homozygous for methionine (129MM) or heterozygous for methionine/valine (129MV). Methods Fifteen affected subjects with 129MM, 129MV, and 129VV underwent comparative evaluation at the National Prion Disease Pathology Surveillance Center for clinical, histopathologic, immunohistochemical, genotypical, and PrP characteristics. Results Disease duration (between 22 and 45 months) was significantly different in the 129VV and 129MV subjects. Most other phenotypic features along with the PrP electrophoretic profile were similar but distinguishable in the 3 129 genotypes. A major difference laid in the sensitivity to protease digestion of the disease-associated PrP, which was high in 129VV but much lower, or altogether lacking, in 129MV and 129MM. This difference prompted the substitution of the original designation with “variably protease-sensitive prionopathy” (VPSPr). None of the subjects had mutations in the PrP gene coding region. Interpretation Because all 3 129 genotypes are involved, and are associated with distinguishable phenotypes, VPSPr becomes the second sporadic prion protein disease with this feature after Creutzfeldt-Jakob disease, originally reported in 1920. However, the characteristics of the abnormal prion protein suggest that VPSPr is different from typical prion diseases, and perhaps more akin to subtypes of Gerstmann-Sträussler-Scheinker disease. PMID:20695009

Characterization of a protease produced by a Trichoderma harzianum isolate which controls cocoa plant witches' broom disease

PubMed Central

De Marco, Janice L; Felix, Carlos Roberto

2002-01-01

Background Several Trichoderma strains have been reported to be effective in controlling plant diseases, and the action of fungal hydrolytic enzymes has been considered as the main mechanism involved in the antagonistic process. However, although Trichoderma strains were found to impair development of Crinipellis perniciosa, the causal agent of cocoa plant witches' broom disease, no fungal strain is available for effective control of this disease. We have then undertaken a program of construction of hydrolytic enzyme-overproducing Trichoderma strains aiming improvement of the fungal antagonistic capacity. The protease of an indian Trichoderma isolate showing antagonistic activity against C. perniciosa was purified to homogeneity and characterized for its kinetic properties and action on the phytopathogen cell wall. Results A protease produced by the Trichoderma harzianum isolate 1051 was purified to homogeneity by precipitation with ammonium sulfate followed by hydrophobic chromatography. The molecular mass of this protease as determined by SDS-polyacrylamide gel electrophoresis was about 18.8 kDa. Its N-terminal amino acid sequence shares no homology with any other protease. The purified enzyme substantially affected the cell wall of the phytopathogen C. perniciosa. Western-blotting analysis showed that the enzyme was present in the culture supernatant 24 h after the Trichoderma started to grow in casein-containing liquid medium. Conclusions The capacity of the Trichoderma harzianum protease to hydrolyze the cell wall of C. perniciosa indicates that this enzyme may be actually involved in the antagonistic process between the two fungi. This fact strongly suggest that hydrolytic enzyme over-producing transgenic fungi may show superior biocontrol capacity. PMID:11835696

Crystal Structures of the Histo-Aspartic Protease (HAP) from Plasmodium falciparum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bhaumik, Prasenjit; Xiao, Huogen; Parr, Charity L.

The structures of recombinant histo-aspartic protease (HAP) from malaria-causing parasite Plasmodium falciparum as apoenzyme and in complex with two inhibitors, pepstatin A and KNI-10006, were solved at 2.5-, 3.3-, and 3.05-{angstrom} resolutions, respectively. In the apoenzyme crystals, HAP forms a tight dimer not seen previously in any aspartic protease. The interactions between the monomers affect the conformation of two flexible loops, the functionally important 'flap' (residues 70-83) and its structural equivalent in the C-terminal domain (residues 238-245), as well as the orientation of helix 225-235. The flap is found in an open conformation in the apoenzyme. Unexpectedly, the active sitemore » of the apoenzyme contains a zinc ion tightly bound to His32 and Asp215 from one monomer and to Glu278A from the other monomer, with the coordination of Zn resembling that seen in metalloproteases. The flap is closed in the structure of the pepstatin A complex, whereas it is open in the complex with KNI-10006. Although the binding mode of pepstatin A is significantly different from that in other pepsin-like aspartic proteases, its location in the active site makes unlikely the previously proposed hypothesis that HAP is a serine protease. The binding mode of KNI-10006 is unusual compared with the binding of other inhibitors from the KNI series to aspartic proteases. The novel features of the HAP active site could facilitate design of specific inhibitors used in the development of antimalarial drugs.« less

Neuropathogenic Escherichia coli K1 does not exhibit proteolytic activities to exert its pathogenicity.

PubMed

Iqbal, Junaid; Rajani, Mehak; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

2013-05-01

Proteases are well-known virulence factors that promote survival, pathogenesis and immune evasion of many pathogens. Several lines of evidence suggest that the blood-brain barrier permeability is a prerequisite in microbial invasion of the central nervous system. Because proteases are frequently associated with vascular permeability by targeting junctional proteins, here it is hypothesized that neuropathogenic Escherichia coli K1 exhibit proteolytic activities to exert its pathogenicity. Zymographic assays were performed using collagen and gelatin as substrates. The lysates of whole E. coli K1 strain E44, or E. coli K-12 strain HB101 were tested for proteolytic activities. The conditioned media were prepared by incubating bacteria in RPMI-1640 in the presence or absence of serum. The cell-free supernatants were collected and tested for proteases in zymography as mentioned above. Additionally, proteolytic degradation of host immune factors was determined by co-incubating conditioned media with albumin/immunoglobulins using protease assays. When collagen or gelatin were used as substrates in zymographic assays, neither whole bacteria nor conditioned media exhibited proteolytic activities. The conditioned media of neuropathogenic E. coli K1 strain E44, or E. coli K-12 strain HB101 did not affect degradation of albumin and immunoglobulins using protease assays. Neither zymographic assays nor protease assays detected proteolytic activities in either the whole bacteria or conditioned media of E. coli K1 strain E44 and E. coli K-12 strain HB101. These findings suggest that host cell monolayer disruptions and immune evasion strategies are likely independent of proteolytic activities of neuropathogenic E. coli K1.

Human mast cell neutral proteases generate modified LDL particles with increased proteoglycan binding.

PubMed

Maaninka, Katariina; Nguyen, Su Duy; Mäyränpää, Mikko I; Plihtari, Riia; Rajamäki, Kristiina; Lindsberg, Perttu J; Kovanen, Petri T; Öörni, Katariina

2018-04-13

Subendothelial interaction of LDL with extracellular matrix drives atherogenesis. This interaction can be strengthened by proteolytic modification of LDL. Mast cells (MCs) are present in atherosclerotic lesions, and upon activation, they degranulate and release a variety of neutral proteases. Here we studied the ability of MC proteases to cleave apoB-100 of LDL and affect the binding of LDL to proteoglycans. Mature human MCs were differentiated from human peripheral blood-derived CD34 + progenitors in vitro and activated with calcium ionophore to generate MC-conditioned medium. LDL was incubated in the MC-conditioned medium or with individual MC proteases, and the binding of native and modified LDL to isolated human aortic proteoglycans or to human atherosclerotic plaques ex vivo was determined. MC proteases in atherosclerotic human coronary artery lesions were detected by immunofluorescence and qPCR. Activated human MCs released the neutral proteases tryptase, chymase, carboxypeptidase A3, cathepsin G, and granzyme B. Of these, cathepsin G degraded most efficiently apoB-100, induced LDL fusion, and enhanced binding of LDL to isolated human aortic proteoglycans and human atherosclerotic lesions ex vivo. Double immunofluoresence staining of human atherosclerotic coronary arteries for tryptase and cathepsin G indicated that lesional MCs contain cathepsin G. In the lesions, expression of cathepsin G correlated with the expression of tryptase and chymase, but not with that of neutrophil proteinase 3. The present study suggests that cathepsin G in human atherosclerotic lesions is largely derived from MCs and that activated MCs may contribute to atherogenesis by enhancing LDL retention. Copyright © 2018 Elsevier B.V. All rights reserved.

Effekt einer ad libitum verzehrten fettreduzierten Kost, reich an Obst, Gemüse und Milchprodukten auf den Blutdruck bei Borderline-Hypertonikern

NASA Astrophysics Data System (ADS)

Möseneder, Jutta M.

2002-01-01

In der randomisierten, multizentrischen DASH-Studie (Dietary Approaches to Stop Hy-pertension), die unter kontrollierten Bedingungen stattfand, führte eine fettreduzierte Mischkost, reich an Obst, Gemüse und Milchprodukten, bei Borderline-Hypertonikern zu einer signifikanten Blutdrucksenkung. Während der Studienphase wurden Körpermasse, Natrium-Aufnahme sowie Alkoholzufuhr aufgrund der bekannten Einflussnahme auf den Blutdruck konstant gehalten. In der eigenen Pilot-Studie sollte untersucht werden, ob das Ergebnis der DASH-Studie (i) mit deutschen Hypertonikern und (ii) unter habituellen Ernährungs- und Lebensbedingungen mit regelmäßig durchgeführter Ernährungsberatung und ad libitum Verzehr anstelle des streng kontrollierten Studienansatzes bestätigt werden kann. Eine Konstanz der Körpermasse, der Natrium-Urinausscheidung (unter diesem Studienansatz valider als die Aufnahme) und des Alkoholkonsums wurde vorausgesetzt. Die Studienpopulation setzte sich aus 53 übergewichtigen Probanden mit einer nicht medikamentös therapierten Borderline-Hypertonie und ohne Stoffwechselerkrankungen zusammen. Die Studienteilnehmer wurden randomisiert entweder der Idealgruppe mit einer fettarmen Kost reich an Milchprodukten, Obst und Gemüse (ähnlich der DASH-Idealgruppe) oder der Kontrollgruppe mit habitueller Ernährungsweise zugeteilt. Über einen Zeitraum von fünf Wochen wurde den Probanden etwa 50% ihres täglichen Lebensmittelbedarfes entsprechend ihrer Gruppenzugehörigkeit kostenfrei zur Verfügung gestellt. Gelegenheitsblutdruckmessungen und 24h-Blutdruckmessungen, Ernährungs- und Aktivitätsprotokolle, Blut- und Urinproben sowie anthropometrische Messungen wurden vor, während und fünf Wochen nach der Interventionsphase durchgeführt. Die Ergebnisse zeigen, dass in der Idealgruppe keine signifikante Blutdrucksenkung beobachtet werden konnte. Dies lässt sich durch die Tatsache erklären, dass die Lebens-mittel- und Nährstoffaufnahme der deutschen Kontrollgruppe eher der amerikanischen Idealgruppe entsprach. In der Pilot-Studie waren die Unterschiede in der Nährstoffzufuhr zwischen den beiden Gruppen viel geringer als in der DASH-Studie; für eine blutdrucksenkende Ernährungsumstellung bestand somit nur ein geringer Spielraum. Eine weitere Erklärung besteht in der unterschiedlichen Zusammensetzung der Studienpopulation. Bei DASH wurden vorwiegend farbige Probanden (40% höhere Hypertonieprävalenz) untersucht. Die Studienergebnisse lassen also den Schluss zu, dass Ernährungs- und Lebensstilgewohnheiten sowie der genetische Hintergrund der entsprechenden Bevölkerungsgruppe bei der Formulierung von nährstoff- oder lebensmittelbezogenen Empfehlungen zur Senkung des Bluthochdruckes Berücksichtigung finden müssen. The Dietary Approaches to Stop Hypertension trial (DASH), a randomized well-controlled feeding study conducted at 4 medical centers, demonstrated that a low-fat diet, rich in fruits, vegetables and dairy products is able to lower blood pressure of borderline-hypertensive people significantly. Body weight, sodium intake and alcohol consumption were kept constant during the whole study period, due to the known influence on hypertension. Aim of our study project was to investigate whether the results of the DASH study can be confirmed by (i) using a German hypertensive population and (ii) replacing the well-controlled feeding design by allowing ad libitum intake according to dietary ad-vice. The participants were asked to keep their body weight, sodium urine excretion (un-der this study design more valid than sodium intake) and their alcohol consumption con-stant. Our pilot study population consisted of 53 mainly overweight participants with borderline hypertension and without medication or any metabolic disorders. They were randomly assigned to either an ideal diet low in fat and rich in dairy products, fruits and vegetables (similar to the DASH I-group) or a habitual diet as control (C-group). During five weeks the subjects were provided 50 % of their daily intake for free according to their dietary pattern. Single and 24h-blood pressure measurements, dietary weighed and physical activity records, blood and urine samples and anthropometric measurements were collected before, during and five weeks after the intervention period. The study results indicated that no significantly reduction of blood pressure could be observed for both methods in the I-group of our trial. This is due to the fact that the baseline intake of foods and nutrients of the German subjects was nearly corresponding to the intake targeted for the DASH I-group. Therefore, it was impossible to increase the intake of these nutrients in our I-group by the same percentage as in the DASH study. Another explanation may be the different consistency of the study population. About 60% of the DASH study population were African Americans with a known higher prevalence of hy-pertension (about 40%) than Caucasians. The conclusion is that even convincing results of a controlled trial cannot be simply transferred into dietary advice for the general public. For addressing food-based dietary guidelines to reduce the risk of hypertension it is necessary to consider the genetic background as well as the dietary and the lifestyle situation of the target population carefully.

Psychometric Properties of the Persian Version of the Simple Shoulder Test (SST) Questionnaire.

PubMed

Ebrahimzadeh, Mohammad H; Vahedi, Ehsan; Baradaran, Aslan; Birjandinejad, Ali; Seyyed-Hoseinian, Seyyed-Hadi; Bagheri, Farshid; Kachooei, Amir Reza

2016-10-01

To validate the Persian version of the simple shoulder test in patients with shoulder joint problems. Following Beaton`s guideline, translation and back translation was conducted. We reached to a consensus on the Persian version of SST. To test the face validity in a pilot study, the Persian SST was administered to 20 individuals with shoulder joint conditions. We enrolled 148 consecutive patients with shoulder problem to fill the Persian SST, shoulder specific measure including Oxford shoulder score (OSS) and two general measures including DASH and SF-36. To measure the test-retest reliability, 42 patients were randomly asked to fill the Persian-SST for the second time after one week. Cronbach's alpha coefficient was used to demonstrate internal consistency over the 12 items of Persian-SST. ICC for the total questionnaire was 0.61 showing good and acceptable test-retest reliability. ICC for individual items ranged from 0.32 to 0.79. The total Cronbach's alpha was 0.84 showing good internal consistency over the 12 items of the Persian-SST. Validity testing showed strong correlation between SST and OSS and DASH. The correlation with OSS was positive while with DASH scores was negative. The correlation was also good to strong with all physical and most mental subscales of the SF-36. Correlation coefficient was higher with DASH and OSS in compare to SF-36. Persian version of SST found to be valid and reliable instrument for shoulder joint pain and function assessment in Iranian population.

The National Football League Combine: performance differences between drafted and nondrafted players entering the 2004 and 2005 drafts.

PubMed

Sierer, S Patrick; Battaglini, Claudio L; Mihalik, Jason P; Shields, Edgar W; Tomasini, Nathan T

2008-01-01

The purpose of this study was to examine performance differences between drafted and nondrafted athletes (N = 321) during the 2004 and 2005 National Football League (NFL) Combines. We categorized players into one of 3 groups: Skill, Big skill, and Linemen. Skill players (SP) consisted of wide receivers, cornerbacks, free safeties, strong safeties, and running backs. Big skill players (BSP) included fullbacks, linebackers, tight ends, and defensive ends. Linemen (LM) consisted of centers, offensive guards, offensive tackles, and defensive tackles. We analyzed player height and mass, as well as performance on the following combine drills: 40-yard dash, 225-lb bench press test, vertical jump, broad jump, pro-agility shuttle, and the 3-cone drill. Student t-tests compared performance on each of these measures between drafted and nondrafted players. Statistical significance was found between drafted and nondrafted SP for the 40-yard dash (P < 0.001), vertical jump (P = 0.003), pro-agility shuttle (P < 0.001), and 3-cone drill (P < 0.001). Drafted and nondrafted BSP performed differently on the 40-yard dash (P = 0.002) and 3-cone drill (P = 0.005). Finally, drafted LM performed significantly better than nondrafted LM on the 40-yard dash (P = 0.016), 225-lb bench press (P = 0.003), and 3-cone drill (P = 0.005). Certified strength and conditioning specialists will be able to utilize the significant findings to help better prepare athletes as they ready themselves for the NFL Combine.

Pilot randomised trial of a healthy eating behavioural intervention in uncontrolled asthma

PubMed Central

Ma, Jun; Strub, Peg; Lv, Nan; Xiao, Lan; Camargo, Carlos A.; Buist, A. Sonia; Lavori, Philip W.; Wilson, Sandra R.; Nadeau, Kari C.; Rosas, Lisa G.

2016-01-01

Rigorous research on the benefit of healthy eating patterns for asthma control is lacking. We randomised 90 adults with objectively confirmed uncontrolled asthma and a low-quality diet (Dietary Approaches to Stop Hypertension (DASH) scores <6 out of 9) to a 6-month DASH behavioural intervention (n=46) or usual-care control (n=44). Intention-to-treat analyses used repeated-measures mixed models. Participants were middle-aged, 67% female and multiethnic. Compared with controls, intervention participants improved on DASH scores (mean change (95% CI) 0.6 (0, 1.1) versus −0.3 (−0.8, 0.2); difference 0.8 (0.2, 1.5)) and the primary outcome, Asthma Control Questionnaire scores (−0.2 (−0.5, 0) versus 0 (−0.3, 0.3); difference −0.2 (−0.5, 0.1)) at 6 months. The mean group differences in changes in Mini Asthma Quality of Life Questionnaire overall and subdomain scores consistently favoured the intervention over the control group: overall 0.4 (95% CI 0, 0.8), symptoms 0.5 (0, 0.9), environment 0.4 (−0.1, 1.0), emotions 0.4 (−0.2, 0.9) and activities 0.3 (0, 0.7). These differences were modest, but potentially clinical significant. The DASH behavioural intervention improved diet quality with promising clinical benefits for better asthma control and functional status among adults with uncontrolled asthma. A full-scale efficacy trial is warranted. PMID:26493792

Effects of protease and non-starch polysaccharide enzyme on performance, digestive function, activity and gene expression of endogenous enzyme of broilers

PubMed Central

Wang, Mingfa; Zhang, Xiaotu; Wang, Zhixiang

2017-01-01

Three hundred one-day-old male broiler chickens (Ross-308) were fed corn-soybean basal diets containing non-starch polysaccharide (NSP) enzyme and different levels of acid protease from 1 to 42 days of age to investigate the effects of exogenous enzymes on growth performance, digestive function, activity of endogenous digestive enzymes in the pancreas and mRNA expression of pancreatic digestive enzymes. For days 1-42, compared to the control chickens, average daily feed intake (ADFI) and average daily gain (ADG) were significantly enhanced by the addition of NSP enzyme in combination with protease supplementation at 40 or 80 mg/kg (p<0.05). Feed-to-gain ratio (FGR) was significantly improved by supplementation with NSP enzymes or NSP enzyme combined with 40 or 80 mg/kg protease compared to the control diet (p<0.05). Apparent digestibility of crude protein (ADCP) was significantly enhanced by the addition of NSP enzyme or NSP enzyme combined with 40 or 80 mg/kg protease (p<0.05). Cholecystokinin (CCK) level in serum was reduced by 31.39% with NSP enzyme combined with protease supplementation at 160 mg/kg (p<0.05), but the CCK level in serum was increased by 26.51% with NSP enzyme supplementation alone. After 21 days, supplementation with NSP enzyme and NSP enzyme combined with 40 or 80 mg/kg protease increased the activity of pancreatic trypsin by 74.13%, 70.66% and 42.59% (p<0.05), respectively. After 42 days, supplementation with NSP enzyme and NSP enzyme combined with 40 mg/kg protease increased the activity of pancreatic trypsin by 32.45% and 27.41%, respectively (p<0.05). However, supplementation with NSP enzyme and 80 or 160 mg/kg protease decreased the activity of pancreatic trypsin by 10.75% and 25.88%, respectively (p<0.05). The activities of pancreatic lipase and amylase were significantly higher in treated animals than they were in the control group (p<0.05). Supplementation with NSP enzyme, NSP enzyme combined with 40 or 80 mg/kg protease increased pancreatic trypsin mRNA levels by 40%, 44% and 28%, respectively. Supplementation with NSP enzyme and 160 mg/kg protease decreased pancreatic trypsin mRNA levels by 13%. Pancreatic lipase and amylase mRNA expression were significantly elevated in treated animals compared to the control group (p<0.05). These results suggest that the amount of NSP enzyme and acid protease in the diet significantly affects digestive function, endogenous digestive-enzyme activity and mRNA expression in broilers. PMID:28323908

Entamoeba histolytica-secreted cysteine proteases induce IL-8 production in human mast cells via a PAR2-independent mechanism.

PubMed

Lee, Young Ah; Nam, Young Hee; Min, Arim; Kim, Kyeong Ah; Nozaki, Tomoyoshi; Saito-Nakano, Yumiko; Mirelman, David; Shin, Myeong Heon

2014-01-01

Entamoeba histolytica is an extracellular tissue parasite causing colitis and occasional liver abscess in humans. E. histolytica-derived secretory products (SPs) contain large amounts of cysteine proteases (CPs), one of the important amoebic virulence factors. Although tissue-residing mast cells play an important role in the mucosal inflammatory response to this pathogen, it is not known whether the SPs induce mast cell activation. In this study, when human mast cells (HMC-1 cells) were stimulated with SPs collected from pathogenic wild-type amoebae, interleukin IL-8 mRNA expression and production were significantly increased compared with cells incubated with medium alone. Inhibition of CP activity in the SPs with heat or the CP inhibitor E64 resulted in significant reduction of IL-8 production. Moreover, SPs obtained from inhibitors of cysteine protease (ICP)-overexpressing amoebae with low CP activity showed weaker stimulatory effects on IL-8 production than the wild-type control. Preincubation of HMC-1 cells with antibodies to human protease-activated receptor 2 (PAR2) did not affect the SP-induced IL-8 production. These results suggest that cysteine proteases in E. histolytica-derived secretory products stimulate mast cells to produce IL-8 via a PAR2-independent mechanism, which contributes to IL-8-mediated tissue inflammatory responses during the early phase of human amoebiasis. © Y.A. Lee et al., published by EDP Sciences, 2014.

Can HN[double bond, length as m-dash]NH, FN[double bond, length as m-dash]NH, or HN[double bond, length as m-dash]CHOH bridge the σ-hole and the lone pair at P in binary complexes with H2XP, for X = F, Cl, NC, OH, CN, CCH, CH3, and H?

PubMed

Del Bene, Janet E; Alkorta, Ibon; Elguero, José

2015-11-11

Ab initio MP2/aug'-cc-pVTZ calculations have been carried out to investigate the properties of complexes formed between H2XP, for X = F, Cl, NC, OH, CN, CCH, CH3, and H, and the possible bridging molecules HN[double bond, length as m-dash]NH, FN[double bond, length as m-dash]NH, and HN[double bond, length as m-dash]CHOH. H2XP:HNNH and H2XP:FNNH complexes are stabilized by PN pnicogen bonds, except for H2(CH3)P:FNNH and H3P:FNNH which are stabilized by N-HP hydrogen bonds. H2XP:HNCHOH complexes are stabilized by PN pnicogen bonds and nonlinear O-HP hydrogen bonds. For a fixed H2XP molecule, binding energies decrease in the order HNCHOH > HNNH > FNNH, except for the binding energies of H2(CH3)P and H3P with HNNH and FNNH. Binding energies of complexes with HNCHOH and HNNH increase as the P-N1 distance decreases, but binding energies of complexes with FNNH show little dependence on this distance. The large binding energies of H2XP:HNCHOH complexes arise from a cooperative effect involving electron-pair acceptance by P to form a pnicogen bond, and electron-pair donation by P to form a hydrogen bond. The dominant charge-transfer interaction in these complexes involves electron-pair donation by N across the pnicogen bond, except for complexes in which X is one of the more electropositive substituents, CCH, CH3, and H. For these, lone-pair donation by P across the hydrogen bond dominates. AIM and NBO data for these complexes are consistent with their bonding characteristics, showing molecular graphs with bond critical points and charge-transfer interactions associated with hydrogen and pnicogen bonds. EOM-CCSD spin-spin coupling constants (1p)J(P-N) across the pnicogen bond for each series of complexes correlate with the P-N distance. In contrast, (2h)J(O-P) values for complexes H2XP:HNCHOH do not correlate with the O-P distance, a consequence of the nonlinearity of these hydrogen bonds.

Kallikreins - The melting pot of activity and function.

PubMed

Kalinska, Magdalena; Meyer-Hoffert, Ulf; Kantyka, Tomasz; Potempa, Jan

2016-03-01

The human tissue kallikrein and kallikrein-related peptidases (KLKs), encoded by the largest contiguous cluster of protease genes in the human genome, are secreted serine proteases with diverse expression patterns and physiological roles. Because of the broad spectrum of processes that are modulated by kallikreins, these proteases are the subject of extensive investigations. This review brings together basic information about the biochemical properties affecting enzymatic activity, with highlights on post-translational modifications, especially glycosylation. Additionally, we present the current state of knowledge regarding the physiological functions of KLKs in major human organs and outline recent discoveries pertinent to the involvement of kallikreins in cell signaling and in viral infections. Despite the current depth of knowledge of these enzymes, many questions regarding the roles of kallikreins in health and disease remain unanswered. Copyright © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

A novel approach to make homogeneous protease-stable monovalent streptavidin

DOE PAGES

Zhang, M.; Shao, J; Xiao, J.; ...

2015-06-11

The interaction between the tetramer streptavidin and biotin is recognized as one of the strongest non-covalent associations. Owing to the tight and specific binding, the streptavidin-biotin system has been used widely for bimolecular labeling, purification, immobilization, and even for targeted delivery of therapeutics drugs. Here, we report a novel approach to make homogeneous monovalent tetramer streptavidin. The purified monovalent protein showed both thermal stability and protease stability. Unexpectedly, we found that two proteases, Proteinase K (PK) and Subtilisin (SU), can efficiently remove the His8-tag from the wild-type subunit without affecting the tetramer architecture of monovalent streptavidin, thus making it moremore » homogeneous. In addition, crystallization was performed to assure the homogeneity of the monovalent protein prepared. Overall, monovalent streptavidin shows increased homogeneity and will likely be valuable for many future applications in a wide range of research areas.« less

Protease-Activated Receptors and other G-Protein-Coupled Receptors: the Melanoma Connection

PubMed Central

Rosero, Rebecca A.; Villares, Gabriel J.; Bar-Eli, Menashe

2016-01-01

The vast array of G-protein-coupled receptors (GPCRs) play crucial roles in both physiological and pathological processes, including vision, coagulation, inflammation, autophagy, and cell proliferation. GPCRs also affect processes that augment cell proliferation and metastases in many cancers including melanoma. Melanoma is the deadliest form of skin cancer, yet limited therapeutic modalities are available to patients with metastatic melanoma. Studies have found that both chemokine receptors and protease-activated receptors, both of which are GPCRs, are central to the metastatic melanoma phenotype and may serve as potential targets in novel therapies against melanoma and other cancers. PMID:27379162

Protease-Activated Receptors and other G-Protein-Coupled Receptors: the Melanoma Connection.

PubMed

Rosero, Rebecca A; Villares, Gabriel J; Bar-Eli, Menashe

2016-01-01

The vast array of G-protein-coupled receptors (GPCRs) play crucial roles in both physiological and pathological processes, including vision, coagulation, inflammation, autophagy, and cell proliferation. GPCRs also affect processes that augment cell proliferation and metastases in many cancers including melanoma. Melanoma is the deadliest form of skin cancer, yet limited therapeutic modalities are available to patients with metastatic melanoma. Studies have found that both chemokine receptors and protease-activated receptors, both of which are GPCRs, are central to the metastatic melanoma phenotype and may serve as potential targets in novel therapies against melanoma and other cancers.

The Effects of Adult Day Services on Family Caregivers’ Daily Stress, Affect, and Health: Outcomes From the Daily Stress and Health (DaSH) Study

PubMed Central

Zarit, Steven H.; Kim, Kyungmin; Femia, Elia E.; Almeida, David M.; Klein, Laura C.

2014-01-01

Purpose: We examine the effects of use of adult day service (ADS) by caregivers of individuals with dementia (IWD) on daily stressors, affect, and health symptoms. Participants were interviewed for 8 consecutive days. On some days, the IWD attended an ADS program and on the other days caregivers provide most or all of the care at home. Methods: Participants were 173 family caregivers of IWDs using an ADS program. Daily telephone interviews assessed care-related stressors, noncare stressors, positive events, affect, and health symptoms. Multilevel models with data nested within persons were used to examine effects of ADS use on daily stressor exposure, affect, and health symptoms. Results: Caregivers had lower exposure to care-related stressors on ADS days, more positive experiences, and more noncare stressors. ADS use lowered anger and reduced the impact of noncare stressors on depressive symptoms. Implications: The findings demonstrate that stressors on caregivers are partly lowered, and affect is improved on ADS days, which may provide protection against the effects of chronic stress associated with caregiving. PMID:23690056

Terahertz-bandwidth coherence measurements of a quantum dash laser in passive and active mode-locking operation.

PubMed

Martin, Eamonn; Watts, Regan; Bramerie, Laurent; Shen, Alexandre; Gariah, Harry; Blache, Fabrice; Lelarge, Francois; Barry, Liam

2012-12-01

This research carries out coherence measurements of a 42.7 GHz quantum dash (QDash) semiconductor laser when passively, electrically, and optically mode-locked. Coherence of the spectral lines from the mode-locked laser is determined by examining the radio frequency beat-tone linewidth as the mode spacing is increased up to 1.1 THz. Electric-field measurements of the QDash laser are also presented, from which a comparison between experimental results and accepted theory for coherence in passively mode-locked lasers has been performed.

Acceptability of the Current and Proposed Items in the Meal, Ready-to-Eat

DTIC Science & Technology

2002-05-01

Cayenne pepper and Mrs. Dash), and dessert items (Peanut Butter M&Ms, Crispy M&Ms, Pumpkin Pound cake, Almond Poppy Pound cake, and a Chocolate Mint...Almost Every Entree 16.34 Every Entree 22.18 Because the Cayenne pepper and Mrs. Dash are not included in the current MREs, the soldiers were asked... pepper . Half(54.47%) wish to have Hot sauce in ’·every/almost every’· entree (Table 3). Forty percent (43.29%) would like to have Cayenne pepper "never

Results of surgical treatment of acromioclavicular dislocations type III using modified Weaver Dunn technique.

PubMed

López-Alameda, S; Fernández-Santás, T; García-Villanueva, A; Varillas-Delgado, D; Garcia de Lucas, F

To evaluate the clinical and radiological results of the surgical treatment of type III acromioclavicular dislocations using the Weaver-Dunn technique in the delayed phase. A non-randomised controlled retrospective observational study of 38 patients operated between January 2006 and December 2014. We excluded 10 patients due to death or non-localisation. We collected demographic data, time to intervention, complications, analysing the Visual Analog Scale, DASH and Oxford Shoulder Score and the updated radiological result. mean age of patients with right-dominant shoulder affected in 71% of cases predominantly by non-level falls was 35. 70% of the cases had subjective perception of both recovery of strength and disappearance of deformity. Full radiological reduction was observed in 95% of the cases with the appearance of mild osteoarthritis in 44% and moderate osteoarthritis in 5.6%. The results of the DASH presented values of 12,939 (±16,851) and the OSS of 42,736 (±7,794), indicating satisfactory articular function. The data from this study shows similar results to previous studies regarding subjective recovery of strength, maintenance of anatomical reduction, functional test results and efficacy of the Weaver-Dunn technique. The modified Weaver-Dunn technique provided good clinical and radiological results with patient reincorporation to their usual activities and maintenance over time. Copyright © 2017 SECOT. Publicado por Elsevier España, S.L.U. All rights reserved.

Assessment of simulation fidelity using measurements of piloting technique in flight. II

NASA Technical Reports Server (NTRS)

Ferguson, S. W.; Clement, W. F.; Hoh, R. H.; Cleveland, W. B.

1985-01-01

Two components of the Vertical Motion Simulator (presently being used to assess the fidelity of UH-60A simulation) are evaluated: (1) the dash/quickstop Nap-of-the-earth (NOE) piloting task, and (2) the bop-up task. Data from these two flight test experiments are presented which provide information on the effect of reduced visual field of view, variation in scene content and texture, and the affect of pure time delay in the closed-loop pilot response. In comparison with task performance results obtained in flight tests, the results from the simulation indicate that the pilot's NOE task performance in the simulator is significantly degraded.

Detection of PrP(Sc) in peripheral tissues of clinically affected cattle after oral challenge with bovine spongiform encephalopathy

USDA-ARS?s Scientific Manuscript database

Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative prion disease that affects cattle and can be transmitted to human beings as new variant Creutzfeldt-Jakob disease (vCJD). A protease-resistant, disease-associated isoform of the prion protein (PrP**Sc) accumulates in the central ner...

Regulated Proteolytic Processing of Reelin through Interplay of Tissue Plasminogen Activator (tPA), ADAMTS-4, ADAMTS-5, and Their Modulators

PubMed Central

Krstic, Dimitrije; Rodriguez, Myriam; Knuesel, Irene

2012-01-01

The extracellular signaling protein Reelin, indispensable for proper neuronal migration and cortical layering during development, is also expressed in the adult brain where it modulates synaptic functions. It has been shown that proteolytic processing of Reelin decreases its signaling activity and promotes Reelin aggregation in vitro, and that proteolytic processing is affected in various neurological disorders, including Alzheimer's disease (AD). However, neither the pathophysiological significance of dysregulated Reelin cleavage, nor the involved proteases and their modulators are known. Here we identified the serine protease tissue plasminogen activator (tPA) and two matrix metalloproteinases, ADAMTS-4 and ADAMTS-5, as Reelin cleaving enzymes. Moreover, we assessed the influence of several endogenous protease inhibitors, including tissue inhibitors of metalloproteinases (TIMPs), α-2-Macroglobulin, and multiple serpins, as well as matrix metalloproteinase 9 (MMP-9) on Reelin cleavage, and described their complex interplay in the regulation of this process. Finally, we could demonstrate that in the murine hippocampus, the expression levels and localization of Reelin proteases largely overlap with that of Reelin. While this pattern remained stable during normal aging, changes in their protein levels coincided with accelerated Reelin aggregation in a mouse model of AD. PMID:23082219

Roles of s3 site residues of nattokinase on its activity and substrate specificity.

PubMed

Wu, Shuming; Feng, Chi; Zhong, Jin; Huan, Liandong

2007-09-01

Nattokinase (Subtilisin NAT, NK) is a bacterial serine protease with high fibrinolytic activity. To probe their roles on protease activity and substrate specificity, three residues of S3 site (Gly(100), Ser(101) and Leu(126)) were mutated by site-directed mutagenesis. Kinetics parameters of 20 mutants were measured using tetrapeptides as substrates, and their fibrinolytic activities were determined by fibrin plate method. Results of mutation analysis showed that Gly(100) and Ser(101) had reverse steric and electrostatic effects. Residues with bulky or positively charged side chains at position 100 decreased the substrate binding and catalytic activity drastically, while residues with the same characters at position 101 could obviously enhance protease and fibrinolytic activity of NK. Mutation of Leu(126) might impair the structure of the active cleft and drastically decreased the activity of NK. Kinetics studies of the mutants showed that S3 residues were crucial to keep protease activity while they moderately affected substrate specificity of NK. The present study provided some original insight into the P3-S3 interaction in NK and other subtilisins, as well as showed successful protein engineering cases to improve NK as a potential therapeutic agent.

A folding-dependent mechanism of antimicrobial peptide resistance to degradation unveiled by solution structure of distinctin

PubMed Central

Raimondo, Domenico; Andreotti, Giuseppina; Saint, Nathalie; Amodeo, Pietro; Renzone, Giovanni; Sanseverino, Marina; Zocchi, Ivana; Molle, Gerard; Motta, Andrea; Scaloni, Andrea

2005-01-01

Many bioactive peptides, presenting an unstructured conformation in aqueous solution, are made resistant to degradation by posttranslational modifications. Here, we describe how molecular oligomerization in aqueous solution can generate a still unknown transport form for amphipathic peptides, which is more compact and resistant to proteases than forms related to any possible monomer. This phenomenon emerged from 3D structure, function, and degradation properties of distinctin, a heterodimeric antimicrobial compound consisting of two peptide chains linked by a disulfide bond. After homodimerization in water, this peptide exhibited a fold consisting of a symmetrical full-parallel four-helix bundle, with a well secluded hydrophobic core and exposed basic residues. This fold significantly stabilizes distinctin against proteases compared with other linear amphipathic peptides, without affecting its antimicrobial, hemolytic, and ion-channel formation properties after membrane interaction. This full-parallel helical orientation represents a perfect compromise between formation of a stable structure in water and requirement of a drastic structural rearrangement in membranes to elicit antimicrobial potential. Thus, distinctin can be claimed as a prototype of a previously unrecognized class of antimicrobial derivatives. These results suggest a critical revision of the role of peptide oligomerization whenever solubility or resistance to proteases is known to affect biological properties. PMID:15840728

Factors affecting decay of Salmonella Birkenhead and coliphage MS2 during mesophilic anaerobic digestion and air drying of sewage sludge.

PubMed

Mondal, Tania; Rouch, Duncan A; Thurbon, Nerida; Smith, Stephen R; Deighton, Margaret A

2015-06-01

Factors affecting the decay of Salmonella Birkenhead and coliphage, as representatives of bacterial and viral pathogens, respectively, during mesophilic anaerobic digestion (MAD) and air drying treatment of anaerobically digested sewage sludge were investigated. Controlled concentrations of S. Birkenhead were inoculated into non-sterile, autoclaved, γ-irradiated and nutrient-supplemented sludge and cultures were incubated at 37 °C (MAD sludge treatment temperature) or 20 °C (summer air drying sludge treatment temperature). Nutrient limitation caused by microbial competition was the principal mechanism responsible for the decay of S. Birkenhead by MAD and during air drying of digested sludge. The effects of protease activity in sludge on MS2 coliphage decay in digested and air dried sludge were also investigated. MS2 coliphage showed a 3.0-3.5 log10 reduction during incubation with sludge-protease extracts at 37 °C for 25 h. Proteases produced by indigenous microbes in sludge potentially increase coliphage inactivation and may therefore have a significant role in the decay of enteric viruses in sewage sludge. The results help to explain the loss of viability of enteric bacteria and viral pathogens with treatment process time and contribute to fundamental understanding of the various biotic inactivation mechanisms operating in sludge treatment processes at mesophilic and ambient temperatures.

Activity-based probes for the ubiquitin conjugation-deconjugation machinery: new chemistries, new tools, and new insights.

PubMed

Hewings, David S; Flygare, John A; Bogyo, Matthew; Wertz, Ingrid E

2017-05-01

The reversible post-translational modification of proteins by ubiquitin and ubiquitin-like proteins regulates almost all cellular processes, by affecting protein degradation, localization, and complex formation. Deubiquitinases (DUBs) are proteases that remove ubiquitin modifications or cleave ubiquitin chains. Most DUBs are cysteine proteases, which makes them well suited for study by activity-based probes. These DUB probes report on deubiquitinase activity by reacting covalently with the active site in an enzyme-catalyzed manner. They have proven to be important tools to study DUB selectivity and proteolytic activity in different settings, to identify novel DUBs, and to characterize deubiquitinase inhibitors. Inspired by the efficacy of activity-based probes for DUBs, several groups have recently reported probes for the ubiquitin conjugation machinery (E1, E2, and E3 enzymes). Many of these enzymes, while not proteases, also posses active site cysteine residues and can be targeted by covalent probes. In this review, we will discuss how features of the probe (cysteine-reactive group, recognition element, and reporter tag) affect reactivity and suitability for certain experimental applications. We will also review the diverse applications of the current probes, and discuss the need for new probe types to study emerging aspects of ubiquitin biology. © 2017 Federation of European Biochemical Societies.

Purification and characterization of a protease produced by Bacillus megaterium RRM2: application in detergent and dehairing industries.

PubMed

Rajkumar, Renganathan; Jayappriyan, Kothilmozhian Ranishree; Rengasamy, Ramasamy

2011-12-01

An alkaline serine protease produced by Bacillus megaterium RRM2 isolated from the red alga, Kappaphycus alvarezii (Doty) Doty ex Silva was studied for the first time and the same analyzed for the production of protease in the present study. Identification of the bacterium was done on the basis of both biochemical analysis and by 16S rDNA sequence analysis. The extracellular protease obtained from B. megaterium RRM2 was purified by a three-step process involving ammonium sulphate precipitation, gel filtration (Sephadex G100) and Q-Sepharose column chromatography. The purity was found to be 30.6-fold with a specific activity of 3591.5 U/mg protein with a molecular weight of 27 kDa. The metal ions Ca(2+), Mg(2+), K(+) and Na(+) marginally enhanced the activity of the purified enzyme while Hg(2+), Cu(2+), Fe(2+), CO(2+) and Zn(2+), had reduced the activity. The enzyme was found to be active in the pH range of 9.0-10.0 and remained active up to 60 °C. Phenyl Methyl Sulfonyl Fluoride (PMSF) inhibited the enzyme activity, thus, confirming that this enzyme is an alkaline serine protease. Likewise, DTT also inhibited the enzyme thus confirming the disulfide nature of the enzyme. The enzyme exhibited a high degree of tolerance to Sodium Dodecyl Sulphate (SDS). The partially purified protease when used as an additive in the commercial detergents was found to be a suitable source for washing clothes especially those stained with blood. Further, it showed good dehairing activity within a short duration in goat skin without affecting its collagen component. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Dietary quality indices in relation to cardiometabolic risk among Finnish children aged 6-8 years - The PANIC study.

PubMed

Eloranta, A M; Schwab, U; Venäläinen, T; Kiiskinen, S; Lakka, H M; Laaksonen, D E; Lakka, T A; Lindi, V

2016-09-01

There are no studies on the relationships of dietary quality indices to the clustering of cardiometabolic risk factors in children. We therefore investigated the associations of four dietary quality indices with cardiometabolic risk score and cardiometabolic risk factors in Finnish children. Subjects were a population sample of 204 boys and 198 girls aged 6-8 years. We assessed diet by 4-day food records and calculated Dietary Approaches to Stop Hypertension (DASH) Score, Baltic Sea Diet Score (BSDS), Mediterranean Diet Score (MDS), and Finnish Children Healthy Eating Index (FCHEI). We calculated the age- and sex-adjusted cardiometabolic risk score summing up Z-scores for waist circumference, mean of systolic and diastolic blood pressure and concentrations of fasting serum insulin and fasting plasma glucose, triglycerides and HDL cholesterol, the last multiplying by -1. Higher FCHEI was associated with lower cardiometabolic risk score among boys (standardised regression coefficient β = -0.14, P = 0.044) adjusted for age, physical activity, electronic media time and household income. Higher DASH Score was related to a lower serum insulin in boys (β = -0.15, P = 0.028). Higher DASH Score (β = -0.16, P = 0.023) and FCHEI (β = -0.17, P = 0.014) were related to lower triglyceride concentration in boys. Higher FCHEI was associated with lower triglyceride concentration in girls (β = -0.16, P = 0.033). Higher DASH Score (β = -0.19, P = 0.011) and BSDS (β = -0.23, P = 0.001) were associated with lower plasma HDL cholesterol concentration in girls. Higher FCHEI was associated with lower cardiometabolic risk among boys, whereas DASH Score, BSDS or MDS were not associated with cardiometabolic risk in children. Copyright © 2016 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.

Effects of sodium intake and diet on racial differences in urinary potassium excretion: results from the Dietary Approaches to Stop Hypertension (DASH)-Sodium trial.

PubMed

Turban, Sharon; Thompson, Carol B; Parekh, Rulan S; Appel, Lawrence J

2013-01-01

We previously showed that African Americans excreted less urinary potassium than whites, even while consuming similar diets in the Dietary Approaches to Stop Hypertension (DASH) trial. We hypothesized that a low-sodium diet may eliminate these differences. Data from the DASH-Sodium randomized controlled feeding trial were analyzed. 412 adults with prehypertension or stage 1 hypertension. Random assignment to either a typical American "control" diet (1.7 g [43 mEq] potassium/2,100 kcal/d) or the DASH diet (4.1 g [105 mEq] potassium/2,100 kcal/d). Within each diet, participants received 3 levels of sodium intake in random order for 30 days. 24-hour urine samples were analyzed at the end of each period. The primary outcome was urinary potassium excretion. On the DASH diet, African Americans consistently excreted significantly less urinary potassium (mean 24-hour urinary potassium excretion, 2,594 ± 961 mg [66 ± 25 mEq]) than whites (3,412 ± 1,016 mg [87 ± 26 mEq]) at the highest sodium level; adjusted (P < 0.001); this difference was not altered by sodium level (P = 0.6 comparing white to African American difference in urinary potassium excretion on high- vs low-sodium diet). In contrast, there was a smaller but significant white-African American difference in mean daily urinary potassium excretion in participants fed the control/high-sodium diet that was not present in the control/low-sodium diet (adjusted differences of 281 mg [7 mEq]/d vs 20 mg [0.5 mEq]/d, respectively; P = 0.007). Significant interactions were found between race and diet (P < 0.001) and between race and sodium (P = 0.02). Single rather than multiple urine collections were available at each time. Lack of stool potassium and sweat potassium values. Racial differences in urinary potassium excretion depend on sodium intake and diet. Our results may help explain the previously documented large variability in urinary potassium excretion. Copyright © 2012 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

Comparative effects of different dietary approaches on blood pressure in hypertensive and pre-hypertensive patients: A systematic review and network meta-analysis.

PubMed

Schwingshackl, Lukas; Chaimani, Anna; Schwedhelm, Carolina; Toledo, Estefania; Pünsch, Marina; Hoffmann, Georg; Boeing, Heiner

2018-05-02

Pairwise meta-analyses have shown beneficial effects of individual dietary approaches on blood pressure but their comparative effects have not been established. Therefore we performed a systematic review of different dietary intervention trials and estimated the aggregate blood pressure effects through network meta-analysis including hypertensive and pre-hypertensive patients. PubMed, Cochrane CENTRAL, and Google Scholar were searched until June 2017. The inclusion criteria were defined as follows: i) Randomized trial with a dietary approach; ii) hypertensive and pre-hypertensive adult patients; and iii) minimum intervention period of 12 weeks. In order to determine the pooled effect of each intervention relative to each of the other intervention for both diastolic and systolic blood pressure (SBP and DBP), random effects network meta-analysis was performed. A total of 67 trials comparing 13 dietary approaches (DASH, low-fat, moderate-carbohydrate, high-protein, low-carbohydrate, Mediterranean, Palaeolithic, vegetarian, low-GI/GL, low-sodium, Nordic, Tibetan, and control) enrolling 17,230 participants were included. In the network meta-analysis, the DASH, Mediterranean, low-carbohydrate, Palaeolithic, high-protein, low-glycaemic index, low-sodium, and low-fat dietary approaches were significantly more effective in reducing SBP (-8.73 to -2.32 mmHg) and DBP (-4.85 to -1.27 mmHg) compared to a control diet. According to the SUCRAs, the DASH diet was ranked the most effective dietary approach in reducing SBP (90%) and DBP (91%), followed by the Palaeolithic, and the low-carbohydrate diet (ranked 3rd for SBP) or the Mediterranean diet (ranked 3rd for DBP). For most comparisons, the credibility of evidence was rated very low to moderate, with the exception for the DASH vs. the low-fat dietary approach for which the quality of evidence was rated high. The present network meta-analysis suggests that the DASH dietary approach might be the most effective dietary measure to reduce blood pressure among hypertensive and pre-hypertensive patients based on high quality evidence.

Prospective study of Dietary Approaches to Stop Hypertension– and Mediterranean-style dietary patterns and age-related cognitive change: the Cache County Study on Memory, Health and Aging123

PubMed Central

Munger, Ronald G; Cutler, Adele; Quach, Anna; Bowles, Austin; Corcoran, Christopher; Tschanz, JoAnn T; Norton, Maria C; Welsh-Bohmer, Kathleen A

2013-01-01

Background: Healthy dietary patterns may protect against age-related cognitive decline, but results of studies have been inconsistent. Objective: We examined associations between Dietary Approaches to Stop Hypertension (DASH)– and Mediterranean-style dietary patterns and age-related cognitive change in a prospective, population-based study. Design: Participants included 3831 men and women ≥65 y of age who were residents of Cache County, UT, in 1995. Cognitive function was assessed by using the Modified Mini-Mental State Examination (3MS) ≤4 times over 11 y. Diet-adherence scores were computed by summing across the energy-adjusted rank-order of individual food and nutrient components and categorizing participants into quintiles of the distribution of the diet accordance score. Mixed-effects repeated-measures models were used to examine 3MS scores over time across increasing quintiles of dietary accordance scores and individual food components that comprised each score. Results: The range of rank-order DASH and Mediterranean diet scores was 1661–25,596 and 2407–26,947, respectively. Higher DASH and Mediterranean diet scores were associated with higher average 3MS scores. People in quintile 5 of DASH averaged 0.97 points higher than those in quintile 1 (P = 0.001). The corresponding difference for Mediterranean quintiles was 0.94 (P = 0.001). These differences were consistent over 11 y. Higher intakes of whole grains and nuts and legumes were also associated with higher average 3MS scores [mean quintile 5 compared with 1 differences: 1.19 (P < 0.001), 1.22 (P < 0.001), respectively]. Conclusions: Higher levels of accordance with both the DASH and Mediterranean dietary patterns were associated with consistently higher levels of cognitive function in elderly men and women over an 11-y period. Whole grains and nuts and legumes were positively associated with higher cognitive functions and may be core neuroprotective foods common to various healthy plant-centered diets around the globe. PMID:24047922

Endoscopic repair of tears of the superficial layer of the distal triceps tendon.

PubMed

Heikenfeld, Roderich; Listringhaus, Rico; Godolias, Georgios

2014-07-01

The purpose of this study was to evaluate the results after endoscopic repair of partial superficial layer triceps tendon tears. Fourteen patients treated surgically between July 2005 and December 2012 were studied prospectively for 12 months. Indication for surgery was a partial detachment of the triceps tendon from the olecranon that was proved by magnetic resonance imaging (MRI) in all cases. Ten of these patients had chronic olecranon bursitis. All patients were treated with endoscopic surgery including bursectomy and repair of the distal triceps tendon with double-loaded suture anchors. Clinical examination of the patients as well as functional and subjective scores (Mayo Elbow Performance Index [MEPI], Disabilities of the Arm, Shoulder and Hand Score [Quick DASH]) were obtained preoperatively and postoperatively at 6 and 12 months. An isokinetic strength measurement and MRI were performed preoperatively and 12 months after surgery. All 14 patients were completely evaluated. The MEPI and Quick DASH Score improved significantly after the repair at all postoperative examinations. The MEPI gained 29 points, up to 96 points at last follow-up (P < .05), and the Quick DASH Score went down 15.6 points after 12 months to 4.5 points (P < .05). Maximum extension power improved 55.8%, up to 94.7% at last follow-up compared with the contralateral side. Using MRI, we found one reruptured partial tear of the triceps tendon that did not require revision surgery. Although triceps tendon ruptures are generally uncommon, partial superficial tears might be more common than previously described. Once the diagnosis is made, endoscopic repair is a method leading to good clinical results with improved function of the affected elbow. Endoscopic repair of superficial tears of the triceps tendon is able to restore function and strength and leads to excellent clinical results after 1 year. Strength recovers to nearly that of the contralateral side, and serious complications appear to be infrequent. Level IV, therapeutic case series. Copyright © 2014 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.

Functional measures developed for clinical populations identified impairment among active workers with upper extremity disorders

PubMed Central

Gardner, Bethany T.; Dale, Ann Marie; Buckner-Petty, Skye; Rachford, Robert; Strickland, Jaime; Kaskutas, Vicki; Evanoff, Bradley

2017-01-01

Purpose Few studies have explored measures of function across a range of health outcomes in a general working population. Using four upper extremity (UE) case definitions from the scientific literature, we described the performance of functional measures of work, activities of daily living, and overall health. Methods A sample of 573 workers completed several functional measures: modified recall versions of the QuickDASH, Levine Functional Status Scale (FSS), DASH Work module (DASH-W), and standard SF-8 physical component score. We determined case status based on four UE case definitions: 1) UE symptoms, 2) UE musculoskeletal disorders (MSD), 3) carpal tunnel syndrome (CTS), and 4) work limitations due to UE symptoms. We calculated effect sizes for each case definition to show the magnitude of the differences that were detected between cases and non-cases for each case definition on each functional measure. Sensitivity and specificity analyses showed how well each measure identified functional impairments across the UE case definitions. Results All measures discriminated between cases and non-cases for each case definition with the largest effect sizes for CTS and work limitations, particularly for the modified FSS and DASH-W measures. Specificity was high and sensitivity was low for outcomes of UE symptoms and UE MSD in all measures. Sensitivity was high for CTS and work limitations. Conclusions Functional measures developed specifically for use in clinical, treatment-seeking populations may identify mild levels of impairment in relatively healthy, active working populations, but measures performed better among workers with CTS or those reporting limitations at work. PMID:26091980

Reaction of the thermo-labile triazenide Na[tBu3SiNNNSiMe3] with CO2: formation of the imido carbonate (tBu3SiO)(Me3SiO)C[double bond, length as m-dash]N-SitBu3 and carbamine acid (tBu3SiO)CONH2.

PubMed

Lerner, H-W; Bolte, M; Wagner, M

2017-07-11

The thermo-labile triazenide Na[tBu 3 SiNNNSiMe 3 ] was prepared by the reaction of Me 3 SiN 3 with Na(thf) 2 [SitBu 3 ] in pentane at -78 °C. Treatment of Na[tBu 3 SiNNNSiMe 3 ] with an excess of carbon dioxide in pentane at -78 °C yielded the imido carbonate (tBu 3 SiO)(Me 3 SiO)C[double bond, length as m-dash]N-SitBu 3 and the carbamine acid (tBu 3 SiO)CONH 2 along with other products. From the reaction solution we could isolate the imido carbonate (tBu 3 SiO)(Me 3 SiO)C[double bond, length as m-dash]N-SitBu 3 and carbamine acid (tBu 3 SiO)CONH 2 . At first single crystals of the carbamine acid (tBu 3 SiO)CONH 2 (triclinic, space group P1[combining macron]) were grown from this solution at room temperature. A second crop of crystals were obtained by concentrating the solution. The second charge consisted of the imido carbonate (tBu 3 SiO)(Me 3 SiO)C[double bond, length as m-dash]N-SitBu 3 (monoclinic, space group P2 1 /n).

Association Between Blood Pressure and Adherence to French Dietary Guidelines.

PubMed

Lelong, Helene; Blacher, Jacques; Menai, Mehdi; Galan, Pilar; Fezeu, Leopold; Hercberg, Serge; Kesse-Guyot, Emmanuelle

2016-08-01

Adopting a healthy diet like the Dietary Approach to Stop Hypertension (DASH) or Mediterranean diet (MD) represents a major lifestyle for blood pressure (BP) control in general population. Nutritional policies, such as the French Nutrition and Health Program (Programme National Nutrition Santé or PNNS), have been implemented in several countries with the aims of preventing chronic diseases. The objective of our study was to investigate association between BP and adherence to PNNS guidelines compared with adherence to DASH or MD. We conducted a cross-sectional study in 11,302 untreated participants from the NutriNet-Santé study, a French web-based cohort study. Three validated scores reflecting adherence to PNNS guidelines, DASH diet, and MD were calculated from repeated 24-hour records. Three BP measurements using a standardized protocol were collected. Multivariate linear models were used to assess the associations between the dietary scores and BP. In women, independent of age, socioeconomic status, body mass index, tobacco use, alcohol consumption, and physical activity, adherence to PNNS guidelines was inversely significantly associated with systolic BP (β = -0.63, P < 0.0001). This association was of similar amplitude that between BP and adherence to DASH-style diet (β = -0.66, P < 0.0001) or MD (β = -0.63, P = 0.0002). No significant association was found in men. Adherence to French nutritional recommendations was found negatively associated with BP at the same magnitude as adherence to well-known dietary pattern in the prevention and treatment of hypertension. © American Journal of Hypertension, Ltd 2016. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Acceptability and feasibility of the 'DASH for Asthma' intervention in a randomized controlled trial pilot study.

PubMed

Blonstein, Andrea C; Lv, Nan; Camargo, Carlos A; Wilson, Sandra R; Buist, A Sonia; Rosas, Lisa G; Strub, Peg; Ma, Jun

2016-08-01

'DASH for Asthma' (n 90) was a 6-month randomized controlled trial that demonstrated potential benefits of a DASH (Dietary Approaches to Stop Hypertension) behavioural intervention for improving diet quality and asthma control by comparing intervention to usual care in adults with uncontrolled asthma. The present study examined acceptability and feasibility of the intervention from the perspective of intervention participants and lifestyle coaches. Grounded in Social Cognitive Theory, the 3-month intensive stage, including three individual and eight group sessions, focused on diet modifications and behavioural self-regulation. The 3-month maintenance stage contained telephone consultations. Participants and lifestyle coaches completed surveys including 5-point Likert scales and open-ended questions. We analysed data using descriptive and inductive content analyses. Forty-six intervention participants (survey response rate was 65-72 %) and two lifestyle coaches. Participants and lifestyle coaches were highly satisfied (all mean ratings >4) with individual and group sessions. Participants identified mastery of knowledge and skills (awareness, goal setting, self-monitoring, problem solving), social learning (class members sharing experiences and ideas) and good coaching skills (reflective listening, empathy, motivational counselling) as important contributors to self-efficacy and programme satisfaction. Participants also valued personalized feedback received in individual sessions. Lifestyle coaches viewed participant engagement as a facilitator to effective sessions. Finally, participants and lifestyle coaches identified food tasting as beneficial for observational learning and facilitation of participant engagement. High class attendance and self-monitoring rate also reflected the high engagement among participants. The DASH behavioural intervention was feasible and highly acceptable to participants with uncontrolled asthma and lifestyle coaches.

Insertion of terminal alkyne into Pt-N bond of the square planar [PtI2(Me2phen)] complex.

PubMed

Benedetti, Michele; De Castro, Federica; Lamacchia, Vincenza; Pacifico, Concetta; Natile, Giovanni; Fanizzi, Francesco P

2017-11-21

The reactivity of [PtX 2 (Me 2 phen)] complexes (X = Cl, Br, I; Me 2 phen = 2,9-dimethyl-1,10-phenanthroline) with terminal alkynes has been investigated. Although the dichlorido species [PtCl 2 (Me 2 phen)] exhibits negligible reactivity, the bromido and iodido derivatives lead in short time to the formation of five-coordinate Pt(ii) complexes of the type [PtX 2 (Me 2 phen)(η 2 -CH[triple bond, length as m-dash]CR)] (X = Br, I; R = Ph, n-Bu), in equilibrium with the starting reagents. Similar to analogous complexes with simple acetylene, the five coordinate species can also undergo dissociation of an halido ligand and formation of the transient square-planar cationic species [PtX(Me 2 phen)(η 2 -CH[triple bond, length as m-dash]CR)] + . This latter can further evolve to give an unusual, sparingly soluble square planar product where the former terminal alkyne is converted into a :C[double bond, length as m-dash]C(H)(R) moiety with the α-carbon bridging the Pt(ii) core with one of the two N-donors of coordinated Me 2 phen. The final product [PtX 2 {κ 2 -N,C-(Z)-N[combining low line]1-N10-C[combining low line][double bond, length as m-dash]C(H)(R)}] (N1-N10 = 2,9-dimethyl-1,10-phenanthroline; X = Br, I) contains a Pt-N-C-C-N-C six-membered chelate ring in a square planar Pt(ii) coordination environment.

Heterotrimeric G protein alpha subunit controls growth, stress response, extracellular protease activity, and cyclopiazonic acid production in Penicillium camemberti.

PubMed

García-Rico, Ramón O; Gil-Durán, Carlos; Rojas-Aedo, Juan F; Vaca, Inmaculada; Figueroa, Luis; Levicán, Gloria; Chávez, Renato

2017-09-01

The fungus Penicillium camemberti is widely used in the ripening of various bloomy-rind cheeses. Several properties of P. camemberti are important in cheese ripening, including conidiation, growth and enzyme production, among others. However, the production of mycotoxins such as cyclopiazonic acid during the ripening process by P. camemberti has raised concerns among consumers that demand food with minimal contamination. Here we show that overexpressing an α-subunit from the subgroup I of the heterotrimeric G protein (Gαi) influences several of these processes: it negatively affects growth in a media-dependent manner, triggers conidial germination, reduces the rate of sporulation, affects thermal and osmotic stress resistance, and also extracellular protease and cyclopiazonic acid production. Our results contribute to understanding the biological determinants underlying these biological processes in the economically important fungus P. camemberti. Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Metabolic effects of the HIV protease inhibitor--saquinavir in differentiating human preadipocytes.

PubMed

Bociąga-Jasik, Monika; Polus, Anna; Góralska, Joanna; Czech, Urszula; Gruca, Anna; Śliwa, Agnieszka; Garlicki, Aleksander; Mach, Tomasz; Dembińska-Kieć, Aldona

2013-01-01

The iatrogenic, HIV-related lipodystrophy is associated with development of the significant metabolic and cardiovascular complications. The underlying mechanisms of antiretroviral (ARV) drugs are not completely explored. The aim of the study was to characterize effects of the protease inhibitor (PI)--saquinavir (SQV) on metabolic functions, and gene expression during differentiation in cells (Chub-S7) culture. SQV in concentrations observed during antiretroviral therapy (ART) significantly decreased mitochondrial membrane potential (MMP), oxygen consumption and ATP generation. The effects were greater in already differentiated cells. This was accompanied by characteristic changes in the expression of the genes involved in endoplasmic reticulum (ER) stress, and differentiation (lipid droplet formation) process such as: WNT10a, C/EBPa, AFT4, CIDEC, ADIPOQ, LPIN1. The results indicate that SQV affects not only metabolic (mitochondrial) activity of adipocytes, but affects the expression of genes related to differentiation and to a lesser extent to cell apoptosis.

Secure distributed genome analysis for GWAS and sequence comparison computation.

PubMed

Zhang, Yihua; Blanton, Marina; Almashaqbeh, Ghada

2015-01-01

The rapid increase in the availability and volume of genomic data makes significant advances in biomedical research possible, but sharing of genomic data poses challenges due to the highly sensitive nature of such data. To address the challenges, a competition for secure distributed processing of genomic data was organized by the iDASH research center. In this work we propose techniques for securing computation with real-life genomic data for minor allele frequency and chi-squared statistics computation, as well as distance computation between two genomic sequences, as specified by the iDASH competition tasks. We put forward novel optimizations, including a generalization of a version of mergesort, which might be of independent interest. We provide implementation results of our techniques based on secret sharing that demonstrate practicality of the suggested protocols and also report on performance improvements due to our optimization techniques. This work describes our techniques, findings, and experimental results developed and obtained as part of iDASH 2015 research competition to secure real-life genomic computations and shows feasibility of securely computing with genomic data in practice.

Secure distributed genome analysis for GWAS and sequence comparison computation

PubMed Central

2015-01-01

Background The rapid increase in the availability and volume of genomic data makes significant advances in biomedical research possible, but sharing of genomic data poses challenges due to the highly sensitive nature of such data. To address the challenges, a competition for secure distributed processing of genomic data was organized by the iDASH research center. Methods In this work we propose techniques for securing computation with real-life genomic data for minor allele frequency and chi-squared statistics computation, as well as distance computation between two genomic sequences, as specified by the iDASH competition tasks. We put forward novel optimizations, including a generalization of a version of mergesort, which might be of independent interest. Results We provide implementation results of our techniques based on secret sharing that demonstrate practicality of the suggested protocols and also report on performance improvements due to our optimization techniques. Conclusions This work describes our techniques, findings, and experimental results developed and obtained as part of iDASH 2015 research competition to secure real-life genomic computations and shows feasibility of securely computing with genomic data in practice. PMID:26733307

DASH, the data and specimen hub of the National Institute of Child Health and Human Development

PubMed Central

Hazra, Rohan; Tenney, Susan; Shlionskaya, Alexandra; Samavedam, Rajni; Baxter, Kristin; Ilekis, John; Weck, Jennifer; Willinger, Marian; Grave, Gilman; Tsilou, Katerina; Songco, David

2018-01-01

The benefits of data sharing are well-established and an increasing number of policies require that data be shared upon publication of the main study findings. As data sharing becomes the new norm, there is a heightened need for additional resources to drive efficient data reuse. This article describes the development and implementation of the Data and Specimen Hub (DASH) by the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) to promote data sharing from NICHD-funded studies and enable researchers to comply with NIH data sharing policies. DASH’s flexible architecture is designed to archive diverse data types and formats from NICHD’s broad scientific portfolio in a manner that promotes FAIR data sharing principles. Performance of DASH over two years since launch is promising: the number of available studies and data requests are growing; three manuscripts have been published from data reanalysis, all within two years of access. Critical success factors included NICHD leadership commitment, stakeholder engagement and close coordination between the governance body and technical team. PMID:29557977

Gain and refractive index dynamics in p-doped InAs quantum dash semiconductor optical amplifiers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Komolibus, Katarzyna; Tyndall National Institute, University College Cork, Cork T12 R5CP; Piwonski, Tomasz, E-mail: tomasz.piwonski@tyndall.ie

The ultrafast carrier dynamics in a p-doped dash-in-a-well structure at 1.5 μm is experimentally investigated. An analysis of the timescales related to carrier relaxation and escape processes as well as the “dynamical” linewidth enhancement factor is presented and compared with results obtained from similar un-doped materials. Intentional p-doping of the active region results in an enhancement of the intermediate timescale of the gain dynamics associated with phonon-assisted electron capture and a reduction of the α-factor due to increased differential gain.

Sonar Based Navigation of an Autonomous Underwater Vehicle

DTIC Science & Technology

1994-06-01

3.1.4 Obstacles that are not on the map are marked, known initial position. 35 Eommd Trejecay of AUV 25 Elied -Solid ine Actual :Dashed line 20, 15 I...actual trajectory. Efifets of Camn*= in do Pai~on of AUV. 20 15- 5- 0 S 10 15 20 25 30 35 Figure 3.16 The effects of currents in the trajectory of the...vehicle. 38 E~med T~MjMMa~ ofAUV 20Eadalmd 3affid one Actual :Dashed hMe 15- 0- 0 5 10 15 20 25 30 35 X (irn faet) Figure 3.17 Position estimation

Virus maturation: dynamics and mechanism of a stabilizing structural transition that leads to infectivity.

PubMed

Steven, Alasdair C; Heymann, J Bernard; Cheng, Naiqian; Trus, Benes L; Conway, James F

2005-04-01

For many viruses, the final stage of assembly involves structural transitions that convert an innocuous precursor particle into an infectious agent. This process -- maturation -- is controlled by proteases that trigger large-scale conformational changes. In this context, protease inhibitor antiviral drugs act by blocking maturation. Recent work has succeeded in determining the folds of representative examples of the five major proteins -- major capsid protein, scaffolding protein, portal, protease and accessory protein -- that are typically involved in capsid assembly. These data provide a framework for detailed mechanistic investigations and elucidation of mutations that affect assembly in various ways. The nature of the conformational change has been elucidated: it entails rigid-body rotations and translations of the arrayed subunits that transfer the interactions between them to different molecular surfaces, accompanied by refolding and redeployment of local motifs. Moreover, it has been possible to visualize maturation at the submolecular level in movies based on time-resolved cryo-electron microscopy.

Tripeptidyl peptidase II promotes fat formation in a conserved fashion.

PubMed

McKay, Renée M; McKay, James P; Suh, Jae Myoung; Avery, Leon; Graff, Jonathan M

2007-12-01

Tripeptidyl peptidase II (TPPII) is a multifunctional and evolutionarily conserved protease. In the mammalian hypothalamus, TPPII has a proposed anti-satiety role affected by degradation of the satiety hormone cholecystokinin 8. Here, we show that TPPII also regulates the metabolic homoeostasis of Caenorhabditis elegans; TPPII RNA interference (RNAi) decreases worm fat stores. However, this occurs independently of feeding behaviour and seems to be a function within fat-storing tissues. In mammalian cell culture, TPPII stimulates adipogenesis and TPPII RNAi blocks adipogenesis. The pro-adipogenic action of TPPII seems to be independent of protease function, as catalytically inactive TPPII also increases adipogenesis. Mice that were homozygous for an insertion in the Tpp2 locus were embryonic lethal. However, Tpp2 heterozygous mutants were lean compared with wild-type littermates, although food intake was normal. These findings indicate that TPPII has central and peripheral roles in regulating metabolism and that TPPII actions in fat-storing tissues might be an ancient function carried out in a protease-independent manner.

Tripeptidyl peptidase II promotes fat formation in a conserved fashion

PubMed Central

McKay, Renée M; McKay, James P; Suh, Jae Myoung; Avery, Leon; Graff, Jonathan M

2007-01-01

Tripeptidyl peptidase II (TPPII) is a multifunctional and evolutionarily conserved protease. In the mammalian hypothalamus, TPPII has a proposed anti-satiety role affected by degradation of the satiety hormone cholecystokinin 8. Here, we show that TPPII also regulates the metabolic homoeostasis of Caenorhabditis elegans; TPPII RNA interference (RNAi) decreases worm fat stores. However, this occurs independently of feeding behaviour and seems to be a function within fat-storing tissues. In mammalian cell culture, TPPII stimulates adipogenesis and TPPII RNAi blocks adipogenesis. The pro-adipogenic action of TPPII seems to be independent of protease function, as catalytically inactive TPPII also increases adipogenesis. Mice that were homozygous for an insertion in the Tpp2 locus were embryonic lethal. However, Tpp2 heterozygous mutants were lean compared with wild-type littermates, although food intake was normal. These findings indicate that TPPII has central and peripheral roles in regulating metabolism and that TPPII actions in fat-storing tissues might be an ancient function carried out in a protease-independent manner. PMID:17932511

Inhibition of Protein Carbamylation in Urea Solution Using Ammonium Containing Buffers

PubMed Central

Sun, Shisheng; Zhou, Jian-Ying; Yang, Weiming; Zhang, Hui

2013-01-01

Urea solution is one of the most commonly employed protein denaturants for protease digestion in proteomic studies. However, it has long been recognized that urea solution can cause carbamylation at the N-termini of proteins/peptides and at the side chain amino groups of lysine and arginine residues. Protein/peptide carbamylation blocks protease digestion and affects protein identification and quantification in mass spectrometry analysis by blocking peptide amino groups from isotopic/isobaric labeling and changing peptide charge states, retention times and masses. In addition, protein carbamylation during sample preparation makes it difficult to study in vivo protein carbamylation. In this study, we compared the peptide carbamylation in urea solutions of different buffers and found that ammonium containing buffers were the most effective buffers to inhibit protein carbamylation in urea solution. The possible mechanism of carbamylation inhibition by ammonium containing buffers is discussed, and a revised procedure for the protease digestion of proteins in urea and ammonium containing buffers was developed to facilitate its application in proteomic research. PMID:24161613

Gut pH as a limiting factor for digestive proteolysis in cultured juveniles of the gilthead sea bream (Sparus aurata).

PubMed

Márquez, Lorenzo; Robles, Rocío; Morales, Gabriel A; Moyano, Francisco J

2012-06-01

After the development of the gastric function in juvenile fish, dietary proteins enter a two-phase digestive process comprising an acidic gastric phase followed by an alkaline intestinal phase. However, the main gastric protease, pepsin, is strictly dependent on the existence of a low-enough environmental pH. In 20-g gilthead sea bream, Sparus aurata, the mean minimal gastric pH is close to 4.5, while the mean pH in the duodenal portion of the intestine was nearly fixed at 6.5. The mean maximal gastric content of HCl was approximately 20 microEq for a low-buffering diet. Gastric proteases were more severely affected than intestinal proteases when assayed at actual sub-optimal pH values, 4.5 and 6.5, respectively. When the gastric proteases of juvenile fish were pre-incubated with a citric acid buffer at pH 6.0, the activity at pH 4.5 was very low, whereas when they were pre-incubated with the same buffer at pH 3.0, the activity at pH 4.5 was significantly increased; this fact suggests a deficient activation of zymogens during the gastric digestion and points to a potential approach to improve protein digestion in juvenile gilthead sea bream.

Delivery of Patient-Reported Outcome Instruments by Automated Mobile Phone Text Messaging.

PubMed

Anthony, Christopher A; Lawler, Ericka A; Glass, Natalie A; McDonald, Katelyn; Shah, Apurva S

2017-11-01

Patient-reported outcome (PRO) instruments allow patients to interpret their health and are integral in evaluating orthopedic treatments and outcomes. The purpose of this study was to define: (1) correlation between PROs collected by automated delivery of text messages on mobile phones compared with paper delivery; and (2) patient use characteristics of a technology platform utilizing automated delivery of text messages on mobile phones. Paper versions of the 12-Item Short Form Health Survey (SF-12) and the short form of the Disabilities of the Arm, Shoulder and Hand (QuickDASH) were completed by patients in orthopedic hand and upper extremity clinics. Over the next 48 hours, the same patients also completed the mobile phone portion of the study outside of the clinic which included text message delivery of the SF-12 and QuickDASH, assigned in a random order. Correlations between paper and text message delivery of the 2 PROs were assessed. Among 72 patients, the intraclass correlation coefficient (ICC) between the written and mobile phone delivery of QuickDASH was 0.91 (95% confidence interval [CI], 0.85-0.95). The ICC between the paper and mobile phone delivery of the SF-12 physical health composite score was 0.88 (95% CI, 0.79-0.93) and 0.86 (95% CI, 0.75-0.92) for the SF-12 mental health composite score. We find that text message delivery using mobile phones permits valid assessment of SF-12 and QuickDASH scores. The findings suggest that software-driven automated delivery of text communication to patients via mobile phones may be a valid method to obtain other PRO scores in orthopedic patients.

GeoDash: Assisting Visual Image Interpretation in Collect Earth Online by Leveraging Big Data on Google Earth Engine

NASA Technical Reports Server (NTRS)

Markert, Kel; Ashmall, William; Johnson, Gary; Saah, David; Mollicone, Danilo; Diaz, Alfonso Sanchez-Paus; Anderson, Eric; Flores, Africa; Griffin, Robert

2017-01-01

Collect Earth Online (CEO) is a free and open online implementation of the FAO Collect Earth system for collaboratively collecting environmental data through the visual interpretation of Earth observation imagery. The primary collection mechanism in CEO is human interpretation of land surface characteristics in imagery served via Web Map Services (WMS). However, interpreters may not have enough contextual information to classify samples by only viewing the imagery served via WMS, be they high resolution or otherwise. To assist in the interpretation and collection processes in CEO, SERVIR, a joint NASA-USAID initiative that brings Earth observations to improve environmental decision making in developing countries, developed the GeoDash system, an embedded and critical component of CEO. GeoDash leverages Google Earth Engine (GEE) by allowing users to set up custom browser-based widgets that pull from GEE's massive public data catalog. These widgets can be quick looks of other satellite imagery, time series graphs of environmental variables, and statistics panels of the same. Users can customize widgets with any of GEE's image collections, such as the historical Landsat collection with data available since the 1970s, select date ranges, image stretch parameters, graph characteristics, and create custom layouts, all on-the-fly to support plot interpretation in CEO. This presentation focuses on the implementation and potential applications, including the back-end links to GEE and the user interface with custom widget building. GeoDash takes large data volumes and condenses them into meaningful, relevant information for interpreters. While designed initially with national and global forest resource assessments in mind, the system will complement disaster assessments, agriculture management, project monitoring and evaluation, and more.

Dash Cam videos on YouTube™ offer insights into factors related to moose-vehicle collisions.

PubMed

Rea, Roy V; Johnson, Chris J; Aitken, Daniel A; Child, Kenneth N; Hesse, Gayle

2018-03-26

To gain a better understanding of the dynamics of moose-vehicle collisions, we analyzed 96 videos of moose-vehicle interactions recorded by vehicle dash-mounted cameras (Dash Cams) that had been posted to the video-sharing website YouTube™. Our objective was to determine the effects of road conditions, season and weather, moose behavior, and driver response to actual collisions compared to near misses when the collision was avoided. We identified 11 variables that were consistently observable in each video and that we hypothesized would help to explain a collision or near miss. The most parsimonious logistic regression model contained variables for number of moose, sight time, vehicle slows, and vehicle swerves (AIC c w = 0.529). This model had good predictive accuracy (AUC = 0.860, SE = 0.041). The only statistically significant variable from this model that explained the difference between moose-vehicle collisions and near misses was 'Vehicle slows'. Our results provide no evidence that road surface conditions (dry, wet, ice or snow), roadside habitat type (forested or cleared), the extent to which roadside vegetation was cleared, natural light conditions (overcast, clear, twilight, dark), season (winter, spring and summer, fall), the presence of oncoming traffic, or the direction from which the moose entered the roadway had any influence on whether a motorist collided with a moose. Dash Cam videos posted to YouTube™ provide a unique source of data for road safety planners trying to understand what happens in the moments just before a moose-vehicle collision and how those factors may differ from moose-vehicle encounters that do not result in a collision. Copyright © 2018 Elsevier Ltd. All rights reserved.

A QM/MM study of the initial excited state dynamics of green-absorbing proteorhodopsin.

PubMed

Borin, Veniamin A; Wiebeler, Christian; Schapiro, Igor

2018-04-17

The primary photochemical reaction of the green-absorbing proteorhodopsin is studied by means of a hybrid quantum mechanics/molecular mechanics (QM/MM) approach. The simulations are based on a homology model derived from the blue-absorbing proteorhodopsin crystal structure. The geometry of retinal and the surrounding sidechains in the protein binding pocket were optimized using the QM/MM method. Starting from this geometry the isomerization was studied with a relaxed scan along the C13[double bond, length as m-dash]C14 dihedral. It revealed an "aborted bicycle pedal" mechanism of isomerization that was originally proposed by Warshel for bovine rhodopsin and bacteriorhodopsin. However, the isomerization involved the concerted rotation about C13[double bond, length as m-dash]C14 and C15[double bond, length as m-dash]N, with the latter being highly twisted but not isomerized. Further, the simulation showed an increased steric interaction between the hydrogen at the C14 of the isomerizing bond and the hydroxyl group at the neighbouring tyrosine 200. In addition, we have simulated a nonadiabatic trajectory which showed the timing of the isomerization. In the first 20 fs upon excitation the order of the conjugated double and single bonds is inverted, consecutively the C13[double bond, length as m-dash]C14 rotation is activated for 200 fs until the S1-S0 transition is detected. However, the isomerization is reverted due to the specific interaction with the tyrosine as observed along the relaxed scan calculation. Our simulations indicate that the retinal - tyrosine 200 interaction plays an important role in the outcome of the photoisomerization.

GeoDash: Assisting Visual Image Interpretation in Collect Earth Online by Leveraging Big Data on Google Earth Engine

NASA Astrophysics Data System (ADS)

Markert, K. N.; Ashmall, W.; Johnson, G.; Saah, D. S.; Anderson, E.; Flores Cordova, A. I.; Díaz, A. S. P.; Mollicone, D.; Griffin, R.

2017-12-01

Collect Earth Online (CEO) is a free and open online implementation of the FAO Collect Earth system for collaboratively collecting environmental data through the visual interpretation of Earth observation imagery. The primary collection mechanism in CEO is human interpretation of land surface characteristics in imagery served via Web Map Services (WMS). However, interpreters may not have enough contextual information to classify samples by only viewing the imagery served via WMS, be they high resolution or otherwise. To assist in the interpretation and collection processes in CEO, SERVIR, a joint NASA-USAID initiative that brings Earth observations to improve environmental decision making in developing countries, developed the GeoDash system, an embedded and critical component of CEO. GeoDash leverages Google Earth Engine (GEE) by allowing users to set up custom browser-based widgets that pull from GEE's massive public data catalog. These widgets can be quick looks of other satellite imagery, time series graphs of environmental variables, and statistics panels of the same. Users can customize widgets with any of GEE's image collections, such as the historical Landsat collection with data available since the 1970s, select date ranges, image stretch parameters, graph characteristics, and create custom layouts, all on-the-fly to support plot interpretation in CEO. This presentation focuses on the implementation and potential applications, including the back-end links to GEE and the user interface with custom widget building. GeoDash takes large data volumes and condenses them into meaningful, relevant information for interpreters. While designed initially with national and global forest resource assessments in mind, the system will complement disaster assessments, agriculture management, project monitoring and evaluation, and more.

Rilonacept in the treatment of subacromial bursitis: A randomized, non-inferiority, unblinded study versus triamcinolone acetonide.

PubMed

Carroll, Matthew B; Motley, Spencer A; Wohlford, Susanna; Ramsey, Bryan C

2015-12-01

Subacromial bursitis is caused by inflammation of the bursa that separates the superior surface of the supraspinatus tendon from the overlying coraco-acromial ligament and acromion. While multiple cytokines are implicated, interleukin-1 beta appears to play a prominent role. Rilonacept, an interleukin-1 trap, may be an alternative to corticosteroid injection for the management of this condition. This single center, randomized, non-inferiority, unblinded study recruited 33 subjects over 9 months. Twenty subjects received 160mg intrabursal injection of rilonacept and 13 received a 6mL mixture of lidocaine, bupivacaine, and 80mg triamcinolone acetonide. QuickDASH, subject reported pain, and adverse events were recorded at time of injection, 2 days later, 2 weeks later, and 4 weeks later. Primary outcome was improvement in QuickDASH 4 weeks post-injection. Secondary outcomes were improvement in subject reported pain and occurrence of adverse events at 4 weeks. Both study groups were equally matched for age, gender, ethnicity, and site of bursa injection. Both medications demonstrated a statistically significant improvement in QuickDASH 4 weeks post-injection, but triamcinolone acetonide injection offered greater improvement (P=0.004). Both medications demonstrated improvement in subject reported pain but between group comparison at 4 weeks showed that triamcinolone was superior (P=0.044). No statistically significant differences in adverse events were noted between groups, but subjects who received rilonacept experienced more episodes of diarrhea and headache. While improvement in QuickDASH and pain was noted with a single intrabursal injection of rilonacept at 4 weeks, injection with triamcinolone acetonide was more efficacious. This trial was registered with www.clinicaltrials.gov (NCT01830699). Copyright © 2015 Société française de rhumatologie. Published by Elsevier SAS. All rights reserved.

Long-term functional, subjective and psychological results after single digit replantation.

PubMed

Chen, Jing; Zhang, Ai Xian; Chen, Qing Zhong; Mu, Shuai; Tan, Jun

2018-03-01

The aim of this study was to analyse the long-term functional, subjective, and psychological results after single-digit replantation. Thirty cases of digital replantation (14 thumbs, 12 index fingers, 2 middle fingers, 1 ring finger, and 1 little finger) in 30 patients (7 females and 23 males) with a mean age of 44.2 years (20-65 years) were evaluated at the end of a mean follow-up time of 36 months (19-50 months). The active range of motion of joints, grip and pinch strength, cutaneous sensibility, upper-extremity functioning, and subjective satisfaction were determined using the Disability of Arm, Shoulder, and Hand (DASH) questionnaire and the Michigan Hand Outcomes questionnaire (MHQ). Psychological sequelae, including depression, anxiety, and posttraumatic stress disorder (PTSD), were assessed. A correlation analysis among variables was also performed. The mean score for the DASH questionnaire was 6.6 (range: 0-39.2). The symptom of cold intolerance occurred in 53% of the patients. Two patients were diagnosed with depression, and only one patient exhibited PTSD. The DASH score had a good statistical correlation with total grip strength, pinch grip strength, and static two-point discrimination (S-2PD) (P < 0.05). Several aspects of the MHQ were also statistically relevant to some or all of the three objective results. Furthermore, the grip strength showed significant correlation with DASH and most aspects of the MHQ in multivariate logistic regression analysis (P < 0.05). Total grip strength is the most important factor positively related to subjective outcomes. The incidence rates of psychological symptoms after digit replantation are very low at long-term follow-up. Level IV, therapeutic study. Copyright © 2017 Turkish Association of Orthopaedics and Traumatology. Production and hosting by Elsevier B.V. All rights reserved.

Black breast cancer survivors experience greater upper extremity disability.

PubMed

Dean, Lorraine T; DeMichele, Angela; LeBlanc, Mously; Stephens-Shields, Alisa; Li, Susan Q; Colameco, Chris; Coursey, Morgan; Mao, Jun J

2015-11-01

Over one-third of breast cancer survivors experience upper extremity disability. Black women present with factors associated with greater upper extremity disability, including: increased body mass index (BMI), more advanced disease stage at diagnosis, and varying treatment type compared with Whites. No prior research has evaluated the relationship between race and upper extremity disability using validated tools and controlling for these factors. Data were drawn from a survey study among 610 women with stage I-III hormone receptor positive breast cancer. The disabilities of the arm, shoulder and hand (QuickDASH) is an 11-item self-administered questionnaire that has been validated for breast cancer survivors to assess global upper extremity function over the past 7 days. Linear regression and mediation analysis estimated the relationships between race, BMI and QuickDASH score, adjusting for demographics and treatment types. Black women (n = 98) had 7.3 points higher average QuickDASH scores than White (n = 512) women (p < 0.001). After adjusting for BMI, age, education, cancer treatment, months since diagnosis, and aromatase inhibitor status, Black women had an average 4-point (95 % confidence interval 0.18-8.01) higher QuickDASH score (p = 0.04) than White women. Mediation analysis suggested that BMI attenuated the association between race and disability by 40 %. Even several years post-treatment, Black breast cancer survivors had greater upper extremity disability, which was partially mediated by higher BMIs. Close monitoring of high BMI Black women may be an important step in reducing disparities in cancer survivorship. More research is needed on the relationship between race, BMI, and upper extremity disability.

Relationship between distal radius fracture malunion and arm-related disability: A prospective population-based cohort study with 1-year follow-up

PubMed Central

2011-01-01

Background Distal radius fracture is a common injury and may result in substantial dysfunction and pain. The purpose was to investigate the relationship between distal radius fracture malunion and arm-related disability. Methods The prospective population-based cohort study included 143 consecutive patients above 18 years with an acute distal radius fracture treated with closed reduction and either cast (55 patients) or external and/or percutaneous pin fixation (88 patients). The patients were evaluated with the disabilities of the arm, shoulder and hand (DASH) questionnaire at baseline (concerning disabilities before fracture) and one year after fracture. The 1-year follow-up included the SF-12 health status questionnaire and clinical and radiographic examinations. Patients were classified into three hypothesized severity categories based on fracture malunion; no malunion, malunion involving either dorsal tilt (>10 degrees) or ulnar variance (≥1 mm), and combined malunion involving both dorsal tilt and ulnar variance. Multivariate regression analyses were performed to determine the relationship between the 1-year DASH score and malunion and the relative risk (RR) of obtaining DASH score ≥15 and the number needed to harm (NNH) were calculated. Results The mean DASH score at one year after fracture was significantly higher by a minimum of 10 points with each malunion severity category. The RR for persistent disability was 2.5 if the fracture healed with malunion involving either dorsal tilt or ulnar variance and 3.7 if the fracture healed with combined malunion. The NNH was 2.5 (95% CI 1.8-5.4). Malunion had a statistically significant relationship with worse SF-12 score (physical health) and grip strength. Conclusion Malunion after distal radius fracture was associated with higher arm-related disability regardless of age. PMID:21232088

Proximal fractures of the humerus in patients older than 75 years of age: should we consider operative treatment?

PubMed

de Kruijf, Marjolein; Vroemen, J P A M; de Leur, K; van der Voort, E A M; Vos, D I; Van der Laan, L

2014-06-01

Over 75 % of patients presenting with a proximal humerus fracture are 70 years or older. Very little is known about the outcome after operative treatment of these fractures in very old patients. This study was performed to gain more insight in safety and functional outcome of surgical treatment of proximal humerus fractures in the elderly. In this observational study, we analyzed all operatively treated patients, aged 75 or older, with a proximal humerus fracture between January 2003 and December 2008 in our center. Patient selection was on clinical grounds, based on physical, mental, and social criteria. Complications were evaluated. We used the DASH Questionnaire to investigate functional outcome, pain, and ADL limitations. Sixty-four patients were treated surgically for a displaced proximal fracture of the humerus: 15 two-part, 32 three-part, and 17 four-part fractures. Mean DASH scores were 37.5, 36.9, and 48.6, respectively. Regarding the operative methods, overall good results were obtained with the modern locked plate osteosynthesis (mean DASH 34.4). Prosthetic treatment, mostly used in highly comminuted fractures, often resulted in poor function (mean DASH 72.9). Persistent pain and ADL limitations were more present in more comminuted fractures (64 and 50 % in patients with 4-part fractures vs. 14 % in 2-part fractures). There were no postoperative deaths within 3 months of surgery, and fracture-related and non-fracture-related complication rates were low (non-union 3 %; 1 myocardial infarction). This study shows that it is safe and justifiable to consider surgical treatment of a severely dislocated proximal humerus fracture in selected patients aged 75 and older. According to OCEBM Working Group,Level IV.

Enzymatic activities and effects of mycovirus infection on the virulence of Metarhizium anisopliae in Rhipicephalus microplus.

PubMed

Perinotto, Wendell M S; Golo, Patricia S; Coutinho Rodrigues, Caio J B; Sá, Fillipe A; Santi, Lucélia; Beys da Silva, Walter O; Junges, Angela; Vainstein, Marilene H; Schrank, Augusto; Salles, Cristiane M C; Bittencourt, Vânia R E P

2014-06-16

The present study aimed to evaluate the pathogenic potential of different Metarhizium anisopliae s.l. isolates and to determine whether differences in enzymatic activities of proteases, lipases and chitinases and infection with mycoviruses affect the control of Rhipicephalus microplus achieved by these fungal isolates. Engorged female ticks were exposed to fungal suspensions. The lipolytic and proteolytic activities in the isolates were evaluated using chromogenic substrates and the chitinolytic activity was determined using fluorescent substrates. A gel zymography was performed to determine the approximate size of serine proteases released by M. anisopliae isolates. To detect mycoviral infections, dsRNA was digested using both RNAse A and S1 endonuclease; samples were analyzed on an agarose gel. Four of the five isolates tested were infected with mycovirus; however, the level of control of R. microplus ticks achieved with the only isolate free of infection (isolate CG 347) was low. This finding suggests that mycoviral infection does not affect the virulence of fungi against ticks. Although all five isolates were considered pathogenic to R. microplus, the best tick control and the highest levels of enzymatic activity were achieved with the isolates CG 629 and CG 148. The in vitro activities of lipases, proteases and chitinases produced by M. anisopliae s.l. differed among isolates and may be related to their virulence. Copyright © 2014 Elsevier B.V. All rights reserved.

Effect of protease inhibitors on angiotensin-converting enzyme activity in human T-lymphocytes.

PubMed

Petrov, V; Fagard, R; Lijnen, P

2000-05-01

The purpose of these investigations was to determine whether the aminopeptidase B and leucine aminopeptidase inhibitor bestatin, the chymase inhibitor chymostatin, the calpain inhibitor E-64, and the neutral serine protease inhibitor leupeptin affect the angiotensin converting enzyme (ACE) activity in T-lymphocytes. ACE activity in homogenates of T-lymphocytes or in intact T-lymphocytes in suspension was measured by determining fluorimetrically histidyl-leucine, formed from the conversion of hippuryl-histidyl-leucine, coupled with ophtaldialdehyde. The effect of various concentrations (10(-9) to 10(-3) mol/L) of the angiotensin-converting enzyme inhibitors lisinopril and captopril and of the various protease inhibitors on ACE activity was studied. Lisinopril and captopril reduced the ACE activity in homogenates of T-lymphocytes in a concentration-dependent manner. Lisinopril exhibited a more pronounced inhibition of ACE in T-lymphocytes than did captopril. Chymostatin and E-64 had no effect on the ACE activity in T-lymphocytes, whereas leupeptin inhibited its activity in a dose-dependent fashion. Bestatin, on the contrary, increased the ACE activity in homogenates of T-lymphocytes as well as in intact T-lymphocytes in proportion to the concentration. Our data showed that the ACE activity in T-lymphocytes was stimulated by bestatin and inhibited by leupeptin, whereas chymostatin and E-64 did not affect the ACE activity in T-lymphocytes.

Long-Term Clinical Outcome After Titanium Lunate Arthroplasty for Kienböck Disease.

PubMed

Viljakka, Timo; Tallroth, Kaj; Vastamäki, Martti

2018-04-09

Titanium lunate arthroplasty (TLA) for Kienböck disease was introduced in 1984 to address the silicone-wear particle problem common to silicone lunate implants. We sought to study the outcome of TLA. We identified 11 patients from our hospital database who had undergone TLA between 2001 and 2010. We evaluated pain, range of motion (ROM), function, and radiological outcome at a mean 11 years after surgery. We compared preoperative ROM and radiological findings with final follow-up in the ipsilateral wrist and also made comparisons with the contralateral wrists. No implants were removed, and no wrist joints were fused. Pain on the visual analog scale averaged 0.5 at rest, 0.3 at night, and 2.7 during heavy exertion. Seven patients had no pain at rest and 9 had no pain at night. Range of motion reached 70% of that of the contralateral wrist, and strength reached 81%. The Disabilities of the Arm, Shoulder, and Hand (DASH) score averaged 9.6, optional DASH 9.7, and Mayo wrist score 67.7. Radiologically, only Ståhl and arthrosis indexes differed significantly between affected and unaffected wrists. Two patients had a dorsally dislocated implant, meaning that around 20% of our cases probably meet the criteria for failure. The longer-term results of TLA for stage III Kienböck disease are promising. Therapeutic IV. Copyright © 2018 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.

[Validation of the German version of the Oxford Elbow Score : A cross-sectional study].

PubMed

Marquardt, J; Schöttker-Königer, T; Schäfer, A

2016-08-01

Elbow complaints are complex problems leading to severe consequences for affected people and the healthcare system. The German version of the Oxford Elbow Score (OES) is the first German-speaking instrument that specifically measures elbow complaints from the patient's perspective and changes of their health status. The aim of this study is the validation of the German version of the OES. In this context the internal consistency and the construct validity were investigated. 59 patients with elbow complaints completed the German version of the OES, the DASH and the SF-36 in a cross-sectional study. The internal consistency was calculated with Cronbach's alpha coefficients. Spearman's correlation coefficients were used to confirm construct validity. Cronbach's alpha for pain, function and psychological subscales was 0.88, 0.81 and 0.90, respectively. The whole questionnaire presents a Cronbach's alpha value of 0.93. Convergent construct validity was confirmed with correlation coefficients containing values of -0.84, -0.77 and -0.82 compared to DASH and values ranging from 0.41 to 0.80 compared with the physical domains of the SF-36. The divergent construct validity presented values ranging from 0.07 to 0.20 with the SF-36 domains of "general health perception" and "mental health". The German OES is an internal consistent instrument with good convergent and divergent construct validity. Other aspects of the validity, the reliability and the responsiveness should be confirmed through further studies.

Granular starch hydrolysis for fuel ethanol production

NASA Astrophysics Data System (ADS)

Wang, Ping

Granular starch hydrolyzing enzymes (GSHE) convert starch into fermentable sugars at low temperatures (≤48°C). Use of GSHE in dry grind process can eliminate high temperature requirements during cooking and liquefaction (≥90°C). In this study, GSHE was compared with two combinations of commercial alpha-amylase and glucoamylase (DG1 and DG2, respectively). All three enzyme treatments resulted in comparable ethanol concentrations (between 14.1 to 14.2% v/v at 72 hr), ethanol conversion efficiencies and ethanol and DDGS yields. Sugar profiles for the GSHE treatment were different from DG1 and DG2 treatments, especially for glucose. During simultaneous saccharification and fermentation (SSF), the highest glucose concentration for the GSHE treatment was 7% (w/v); for DG1 and DG2 treatments, maximum glucose concentration was 19% (w/v). GSHE was used in one of the fractionation technologies (enzymatic dry grind) to improve recovery of germ and pericarp fiber prior to fermentation. The enzymatic dry grind process with GSHE was compared with the conventional dry grind process using GSHE with the same process parameters of dry solids content, pH, temperature, time, enzyme and yeast usages. Ethanol concentration (at 72 hr) of the enzymatic process was 15.5% (v/v), which was 9.2% higher than the conventional process (14.2% v/v). Distillers dried grains with solubles (DDGS) generated from the enzymatic process (9.8% db) was 66% less than conventional process (28.3% db). Three additional coproducts, germ 8.0% (db), pericarp fiber 7.7% (db) and endosperm fiber 5.2% (db) were produced. Costs and amounts of GSHE used is an important factor affecting dry grind process economics. Proteases can weaken protein matrix to aid starch release and may reduce GSHE doses. Proteases also can hydrolyze protein into free amino nitrogen (FAN), which can be used as a yeast nutrient during fermentation. Two types of proteases, exoprotease and endoprotease, were studied; protease and urea addition were evaluated in the dry grind process using GSHE (GSH process). Addition of proteases resulted in higher ethanol concentrations (15.2 to 18.0% v/v) and lower (DDGS) yields (32.9 to 45.8% db) compared to the control (no protease addition). As level of proteases and GSHE increased, ethanol concentrations increased and DDGS yields decreased. Proteases addition reduced required GSHE dose. Ethanol concentrations with protease addition alone were higher than with urea or with addition of both protease and urea. Corn endosperm consists of soft and hard endosperm. More exposed starch granules and rough surfaces produced from soft endosperm compared to hard endosperm will create more surface area which will benefit the solid phase hydrolysis as used in GSH process. In this study, the effects of protease, urea, endosperm hardness and GSHE levels on the GSH process were evaluated. Soft and hard endosperm materials were obtained by grinding and sifting flaking grits from dry milling pilot plant. Soft endosperm resulted in higher ethanol concentrations (at 72 hr) compared to ground corn or hard endosperm. Addition of urea increased ethanol concentrations (at 72 hr) for soft and hard endosperm. The effect of protease addition on increasing ethanol concentrations and fermentation rates was more predominant for soft endosperm, less for hard endosperm and least for ground corn. The GSH process with protease resulted in higher ethanol concentration than that with urea. For fermentation of soft endosperm, GSHE dose can be reduced. Ground corn fermented faster at the beginning than hard and soft endosperm due to the presence of inherent nutrients which enhanced yeast growth.

Protease nexin 1 is a potent urinary plasminogen activator inhibitor in the presence of collagen type IV.

PubMed

Crisp, Robert J; Knauer, Mary F; Knauer, Daniel J

2002-12-06

Protease nexin 1 (PN1) in solution forms inhibitory complexes with thrombin or urokinase, which have opposing effects on the blood coagulation cascade. An initial report provided data supporting the idea that PN1 target protease specificity is under the influence of collagen type IV (1). Although collagen type IV demonstrated no effect on the association rate between PN1 and thrombin, the study reported that the association rate between PN1 and urokinase was allosterically reduced 10-fold. This has led to the generally accepted idea that the primary role of PN1 in the brain is to act as a rapid thrombin inhibition and clearance mechanism during trauma and loss of vascular integrity. In studies to identify the structural determinants of PN1 that mediate the allosteric interaction with collagen type IV, we found that protease specificity was only affected after transient exposure of PN1 to acidic conditions that mimic the elution protocol from a monoclonal antibody column. Because PN1 used in previous studies was purified over a monoclonal antibody column, we propose that the allosteric regulation of PN1 target protease specificity by collagen type IV is a result of the purification protocol. We provide both biochemical and kinetic data to support this conclusion. This finding is significant because it implies that PN1 may play a much larger role in the modeling and remodeling of brain tissues during development and is not simply an extravasated thrombin clearance mechanism as previously suggested.

Mechanisms Regulating the Degradation of Dentin Matrices by Endogenous Dentin Proteases and their Role in Dental Adhesion. A Review

PubMed Central

Sabatini, Camila; Pashley, David H.

2015-01-01

Purpose This systematic review provides an overview of the different mechanisms proposed to regulate the degradation of dentin matrices bye host-derived dentin proteases, particularly as it relates to their role in dental adhesion. Methods Significant developments have taken place over the last few years that have contributed to a better understanding of all the factors affecting the durability of adhesive resin restorations. The complexity of dentin-resin interfaces mandates a thorough understanding of all the mechanical, physical and biochemical aspects that play a role in the formation of hybrid layers. The ionic and hydrophilic nature of current dental adhesives yields permeable, unstable hybrid layers susceptible to water sorption, hydrolytic degradation and resin leaching. The hydrolytic activity of host-derived proteases also contributes to the degradation of the resin-dentin bonds. Preservation of the collagen matrix is critical to the improvement of resin-dentin bond durability. Approaches to regulate collagenolytic activity of dentin proteases have been the subject of extensive research in the last few years. A shift has occurred from the use of proteases inhibitors to the use of collagen cross-linking agents. Data provided by fifty-one studies published in peer-reviewed journals between January 1999 and December 2013 was compiled in this systematic review. Results Appraisal of the data provided by the studies included in the present review yielded a summary of the mechanisms which have already proven to be clinically successful and those which need further investigation before new clinical protocols can be adopted. PMID:25831604

Metabolic complications associated with HIV protease inhibitor therapy.

PubMed

Nolan, David

2003-01-01

HIV protease inhibitors were introduced into clinical practice over 7 years ago as an important component of combination antiretroviral drug regimens which in many ways revolutionised the treatment of HIV infection. The significant improvements in prognosis that have resulted from the use of these regimens, combined with the need for lifelong treatment, have increasingly focused attention on the adverse effects of antiretroviral drugs and on the metabolic complications of HIV protease inhibitors in particular. In this review, the cluster of metabolic abnormalities characterised by triglyceride-rich dyslipidaemia and insulin resistance associated with HIV protease inhibitor therapy are considered, along with implications for cardiovascular risk in patients affected by these complications. Toxicity profiles of individual drugs within the HIV protease inhibitor class are examined, as there is an increased recognition of significant intra-class differences both in terms of absolute risk of metabolic complications as well as the particular metabolic phenotype associated with these drugs. Guidelines for clinical assessment and treatment are emphasised, along with pathophysiological mechanisms that may provide a rational basis for the treatment of metabolic complications. Finally, these drug-specific effects are considered within the context of HIV-specific effects on lipid metabolism as well as lifestyle factors that have contributed to a rapidly increasing incidence of similar metabolic syndromes in the general population. These data highlight the importance of individualising patient management in terms of choice of antiretroviral regimen, assessment of metabolic outcomes and use of therapeutic interventions, based on the assessment of baseline (pre-treatment) metabolic status as well as the presence of potentially modifiable cardiovascular risk factors.

Spelling Endings Added to e: Level III, Unit 5, Lesson 1; Capitalization: Lesson 2; Question Marks and Exclamation Points: Lesson 3; Quotation Marks: Lesson 4; Spelling Double Letter Demons: Lesson 5; Colons and Dashes: Lesson 6; Punctuating Series with Commas and Semicolons: Lesson 7; More Confusing Word Pairs: Lesson 8; Separating Sentence Parts with Punctuation: Lesson 9; Other Uses for Commas and Semicolons.

ERIC Educational Resources Information Center

Manpower Administration (DOL), Washington, DC. Job Corps.

This self-study program for the high-school level contains lessons in the following subjects: Spelling Endings Added to e; Capitalization; Question Marks and Exclamation Points; Quotation Marks; Spelling Double Letter Demons; Colons and Dashes; Punctuating Series with Commas and Semicolons; More Confusing Word Pairs; Separating Sentence Parts with…

146-GHz millimeter-wave radio-over-fiber photonic wireless transmission system.

PubMed

Fice, M J; Rouvalis, E; van Dijk, F; Accard, A; Lelarge, F; Renaud, C C; Carpintero, G; Seeds, A J

2012-01-16

We report the experimental implementation of a wireless transmission system with a 146-GHz carrier frequency which is generated by optical heterodyning the two modes from a monolithically integrated quantum dash dual-DFB source. The monolithic structure of the device and the inherent low noise characteristics of quantum dash gain material allow us to demonstrate the transmission of a 1 Gbps ON-OFF keyed data signal with the two wavelengths in a free-running state at 146-GHz carrier wave frequency. The tuning range of the device fully covers the W-band (75 - 110 GHz) and the F-band (90 - 140 GHz).

The coronary artery disease quality dashboard: a chronic care disease management tool in an electronic health record.

PubMed

Jung, Eunice; Schnipper, Jeffrey L; Li, Qi; Linder, Jeffrey A; Rose, Alan F; Li, Ruzhuo; Eskin, Michael S; Housman, Dan; Middleton, Blackford; Einbinder, Jonathan S

2007-10-11

Quality reporting tools, integrated with ambulatory electronic health records (EHRs), may help clinicians understand performance, manage populations, and improve quality. The Coronary Artery Disease Quality Dash board (CAD QD) is a secure web report for performance measurement of a chronic care condition delivered through a central data warehouse and custom-built reporting tool. Pilot evaluation of the CAD Quality Dash board indicates that clinicians prefer a quality report that combines not only structured data from EHRs but one that facilitates actions to be taken on individual patients or on a population, i.e., for case management.

SURGICAL TREATMENT OF DISTAL RADIUS FRACTURES WITH A VOLAR LOCKED PLATE: CORRELATION OF CLINICAL AND RADIOGRAPHIC RESULTS

PubMed Central

Xavier, Claudio Roberto Martins; Dal Molin, Danilo Canesin; dos Santos, Rafael Mota Marins; dos Santos, Roberto Della Torre; Neto, Julio Cezar Ferreira

2015-01-01

Objectives: To analyze and correlate the clinical and radiographic results from patients with distal radius fractures who underwent surgical treatment with a fixed-angle volar locked plate. Methods: Sixty-four patients with distal radius fractures were evaluated. They all underwent surgical treatment with a volar locked plate for the distal radius, with a minimum of six months of postoperative follow-up. They underwent a physical examination that measured range of motion and grip strength, answered the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire and underwent radiographic examination. Results: In the physical examination on the patients, all the range-of-motion measurements were reduced. Grip strength measured in kgf was on average 85.8% of the strength on the unaffected side. The mean DASH score was 15.99. A significant relationship was found between lower DASH scores and losses of extension and grip strength. On the radiographs, the mean values in relation to the unfractured side were 84.0% for radial inclination, 85.4% for radial length and 86.8% for volar deviation of the radius. Loss of radial length was correlated with losses of extension and grip strength. PMID:27027046

Effects of platelet-rich plasma on lateral epicondylitis of the elbow: prospective randomized controlled trial.

PubMed

Palacio, Evandro Pereira; Schiavetti, Rafael Ramos; Kanematsu, Maiara; Ikeda, Tiago Moreno; Mizobuchi, Roberto Ryuiti; Galbiatti, José Antônio

2016-01-01

To evaluate the effects of platelet-rich plasma (PRP) infiltration in patients with lateral epicondylitis of the elbow, through analysis of the Disabilities of the Arm, Shoulder and Hand (DASH) and Patient-Rated Tennis Elbow Evaluation (PRTEE) questionnaires. Sixty patients with lateral epicondylitis of the elbow were prospectively randomized and evaluated after receiving infiltration of three milliliters of PRP, or 0.5% neocaine, or dexamethasone. For the scoring process, the patients were asked to fill out the DASH and PRTEE questionnaires on three occasions: on the day of infiltration and 90 and 180 days afterwards. Around 81.7% of the patients who underwent the treatment presented some improvement of the symptoms. The statistical tests showed that there was evidence that the cure rate was unrelated to the substance applied (p = 0.62). There was also intersection between the confidence intervals of each group, thus demonstrating that the proportions of patients whose symptoms improved were similar in all the groups. At a significance level of 5%, there was no evidence that one treatment was more effective than another, when assessed using the DASH and PRTEE questionnaires.

Joint distraction for thumb carpometacarpal osteoarthritis: a feasibility study with 1-year follow-up.

PubMed

Spaans, Anne J; Minnen, L Paul van; Braakenburg, Assa; Mink van der Molen, Aebele B

2017-08-01

The purpose of this pilot study was to evaluate the feasibility of joint distraction of the first carpometacarpal (CMC1) joint in patients with CMC1 osteoarthritis (OA). An external joint distractor was placed over the CMC1 joint by K-wire fixation in the trapezium and the metacarpal. The joint was distracted 3 mm during surgery. The device was then kept in place for 8 weeks. Disabilities of the Arm, Shoulder, and Hand (DASH) score, Michigan Hand Outcome Questionnaire (MHQ), Visual Analogue Scale (VAS), and grip strength were recorded preoperatively and at set postoperative intervals. Five female patients with an average age of 53 years (range = 41-61) were included. One year postoperatively, average DASH, MHQ, and VAS scores improved compared to preoperative values; DASH 53 to 27, MHQ 48 to 76, and VAS pain 48 to 14. There were no technical problems associated with the device. One patient had a local pin site infection treated successfully with oral antibiotics. This study concludes that joint distraction of the osteoarthritic CMC1 joint is technically feasible. In this small, prospective pilot study the majority of the results were favourable during short-term follow-up.

Can radiological results be surrogate markers of functional outcome in distal radial extra-articular fractures?

PubMed

Kumar, Skand; Penematsa, Subbaraju; Sadri, Maziar; Deshmukh, Subodh C

2008-08-01

The purpose of this study was to evaluate the relationship between radiological and functional results in patients with extra-articular fractures of the distal radius. We conducted a prospective study of radiological and functional assessment in 95 consecutively selected extra-articular distal radius fractures. There were two patient groups: more than 60 and less than 60 years of age. The final fracture union radiographs were analysed for their functional outcome using the Michigan Hand Outcomes Questionnaire (MHQ) and the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire. In patients with acceptable radiological results, 62% (MHQ group) and 72% (DASH group) of patients had satisfactory functional outcome. Analysing patients with satisfactory functional results, 56% (MHQ group) and 59% (DASH group) had satisfactory radiological results. There was a higher proportion of patients with better functional results, despite poor radiological results, in both of the age groups. There was a statistically significant correlation between satisfactory radial tilt and functional outcome in the younger patients. In the older age group, patients with satisfactory radiological results had satisfactory functional outcome (p<0.05). However, in the older age group, satisfactory functional results were achieved, despite unsatisfactory radiological results.

Patient satisfaction and disability after brachial plexus surgery.

PubMed

Kretschmer, Thomas; Ihle, Sarah; Antoniadis, Gregor; Seidel, Julia A; Heinen, Christian; Börm, Wolfgang; Richter, Hans-Peter; König, Ralph

2009-10-01

Little is known about patient satisfaction and disability after brachial plexus surgery. Would patients undergo the procedure again, if they knew the current result beforehand? How do they rate their result and their disability? Of 319 plexus patients who had undergone surgery between 1995 and 2005, 199 received a 65-item questionnaire. Measurement instruments included a new plexus-specific outcome questionnaire (Ulm Questionnaire) with categories of satisfaction, functionality, pain, comorbidities, and work; and the disability of the arm, shoulder, and hand questionnaire (DASH; scale, 0-100). Of 99 returned questionnaires, 70 were returned in a useful form for evaluation. The results of patients with C5-C6 lesions (21 of 70) are as follows: 90% (19 of 21) would undergo surgery again, 95% (20 of 21) were satisfied with the result, and 86% (18 of 21) subjectively improved. The mean DASH score was 41 (standard deviation [SD], 24). The results of patients with C5-C7 lesions (6 of 70) are as follows: 50% (3 of 6) were satisfied and would undergo surgery again, and 67% (4 of 6) improved. The mean DASH score was 46 (SD, 13). The results of patients with C5-T1 lesions (43 of 70) are as follows: 67% (29 of 43) would undergo surgery again, 81% (35 of 42) were satisfied, and 74% (32 of 43) reported improvement. The mean DASH score was 58 (SD, 26). The overall mean DASH score was 52 (SD, 26). Pain since the injury was prevalent in 86% of patients (60 of 70), back pain in 53%, and depression/anxiety in 21%. Fifty-two percent of those who worked before their injury (27 of 53 patients) remained unemployed or incapacitated for work. Forty-five percent of previous workers (24 of 53) returned to their former occupation. Occupational retraining was successful for 70% of patients (16 of 23). The mean duration until return to work was 9 months overall and 5 months for those who returned to their previous occupation. Eighty-seven percent of patients were satisfied with the results and 83% would undergo the procedure again. Despite a high satisfaction rate, patients remained considerably disabled, and half of the previous workers did not return to work. Occupational retraining is effective.

CHRONIC UCL INJURY: A MULTIMODAL APPROACH TO CORRECTING ALTERED MECHANICS AND IMPROVING HEALING IN A COLLEGE ATHLETE— A CASE REPORT

PubMed Central

Patrick, Rachel; McGinty, Josh; Lucado, Ann; Collier, Beth

2016-01-01

ABSTRACT Background Ulnar collateral ligament (UCL) tears and associated Tommy Johns surgical intervention from excessive and poor quality pitching has increased immensely—with more college and professional pitchers undergoing the surgery in 2014 alone than in the 1990s as a whole.1 Faulty mechanics developed at young ages are often well-engrained by the late adolescent years and the minimal healing ability of the largely avascular UCL often leads to delayed safe return to sport.2 Purpose The purpose of this case study was to describe an innovative, multimodal approach to conservative management of a chronic UCL injury in a college-aged baseball pitcher. This innovative approach utilizes both contractile and non-contractile dry needling to enhance soft tissue healing combined with standard conservative treatment to decrease pain and improve sport performance as measured by the Disabilities of Arm, Shoulder and Hand (DASH), Numeric Pain Report Scale (NPRS), and return to sport. Study Design Retrospective Case Report Case Description A collegiate athlete presented to an outpatient orthopedic physical therapy clinic for treatment of UCL sprain approximately six weeks post-injury and platelet-rich plasma injection. Diagnostic testing revealed chronic ligamentous microtrauma. Impairments at evaluation included proximal stabilizing strength deficits, myofascial trigger points throughout the dominant upper extremity, improper pitching form, and inability to pitch in game conditions due to severe pain. Interventions included addressing strength deficits throughout the body, dry needling, and sport-specific biomechanical training with pitching form analysis and correction. Outcomes Conventional DASH and Sport-Specific scale on the DASH and the numeric pain rating scale improved beyond both the minimally clinically important difference and minimal detectable change over the 12 week treatment3,4 At 24-week follow up, conventional DASH scores decreased from 34.20% disability to 3.33% disability while sport-specific DASH scores decreased from 100% disability to 31.25% disability. Although initially unable to compete due to high pain levels, the subject is currently completing his pitching role full-time with 1/10 max pain. Discussion The approach used in this case study provides an innovative approach to conservative UCL partial tear treatment. Dry needling of both contractile and non-contractile tissue in combination with retraining of faulty mechanics may encourage chronically injured ligamentous tissue healing and encourage safe return to sport. Level of evidence Level 4 PMID:27525185

Reliability, validity, and responsiveness of the Japanese version of the Patient-Rated Wrist Evaluation.

PubMed

Imaeda, Toshihiko; Uchiyama, Shigeharu; Wada, Takuro; Okinaga, Shuji; Sawaizumi, Takuya; Omokawa, Shohei; Momose, Toshimitsu; Moritomo, Hisao; Gotani, Hiroyuki; Abe, Yukio; Nishida, Jun; Kanaya, Fuminori

2010-07-01

The Patient-Rated Wrist Evaluation is a regionspecific, self-administered questionnaire consisting of a pain scale (PRWE-P) and a functional scale (PRWE-F), with the latter consisting of specific function (PRWE-SF) and usual function (PRWE-UF). The PRWE was cross-culturally adapted from the original English version by the Impairment Evaluation Committee, Japanese Society for Surgery of the Hand (JSSH). The purpose of this study was to test the reliability, validity, and responsiveness of the Japanese version of PRWE (PRWE-J). A consecutive series of 117 patients with wrist disorders completed the PRWE-J, the JSSH version of the Disabilities of the Arm, Shoulder, and Hand (DASH-JSSH) questionnaire and the 36-Item Short-Form Health Survey (SF-36). Of the 117 patients, 71 were reassessed for test-retest reliability 1 or 2 weeks later. Reliability was investigated by reproducibility and internal consistency. To analyze the validity, a factor analysis (principal axis factoring) of PRWE-J and correlation coefficients between PRWE-J and DASH-JSSH were obtained. Responsiveness was examined by calculating the standardized response mean (SRM) (mean change/SD) and effect size (mean change/SD of baseline value) after open surgery in 50 patients. Cronbach's alpha coefficients for PRWE-P, PRWE-F, and PRWE were 0.90, 0.95, and 0.95, respectively. The intraclass correlation coefficients (ICCs) for the same were 0.86, 0.93, and 0.92, respectively. Unidimensionality of PRWE-P was con-firmed. Bidimensionality of PRWE-F was confirmed and separated clearly into PRWE-SF and PRWE-UF. The correlation coefficients between PRWE-P and PRWE-F or DASH-JSSH were 0.63 or 0.63, respectively. The correlation coefficient between PRWE-F and DASH-JSSH was 0.80. The correlation coefficients between DASH-JSSH and PRWE-SF or PRWE-UF were 0.76 or 0.73, respectively. Moderate correlation was observed in "physical functioning" for SF-36 and PRWE-SF (r = -0.46), PRWE-F (r = -0.46), or PRWE (r = -0.46). The SRMs/effect sizes of PRWE-P, PRWE-F, or PRWE were respectively excellent: 1.7/2.2, 1.2/1.3, 1.6/1.9. The PRWE-J has evaluation capacities equivalent to those of the original PRWE.

Brazilian dietary patterns and the dietary approaches to stop hypertension (DASH) diet-relationship with metabolic syndrome and newly diagnosed diabetes in the ELSA-Brasil study.

PubMed

Drehmer, Michele; Odegaard, Andrew O; Schmidt, Maria Inês; Duncan, Bruce B; Cardoso, Letícia de Oliveira; Matos, Sheila M Alvim; Molina, Maria Del Carmen B; Barreto, Sandhi M; Pereira, Mark A

2017-01-01

Studies evaluating dietary patterns, including the DASH diet, and their relationship with the metabolic syndrome and diabetes may help to understand the role of dairy products (low fat or full fat) in these conditions. Our aim is to identify dietary patterns in Brazilian adults and compare them with the (DASH) diet quality score in terms of their associations with metabolic syndrome and newly diagnosed diabetes in the Brazilian Longitudinal Study of Adult Health-the ELSA-Brasil study. The ELSA-Brasil is a multicenter cohort study comprising 15,105 civil servants, aged 35-74 years at baseline (2008-2010). Standardized interviews and exams were carried out, including an OGTT. We analyzed baseline data for 10,010 subjects. Dietary patterns were derived by principal component analysis. Multivariable logistic regression investigated associations of dietary patterns with metabolic syndrome and newly diagnosed diabetes and multivariable linear regression with components of metabolic syndrome. After controlling for potential confounders, we observed that greater adherence to the Common Brazilian meal pattern (white rice, beans, beer, processed and fresh meats), was associated with higher frequencies of newly diagnosed diabetes, metabolic syndrome and all of its components, except HDL-C. Participants with greater intake of a Common Brazilian fast foods/full fat dairy/milk based desserts pattern presented less newly diagnosed diabetes. An inverse association was also seen between the DASH Diet pattern and the metabolic syndrome, blood pressure and waist circumference. Diet, light foods and beverages/low fat dairy pattern was associated with more prevalence of both outcomes, and higher fasting glucose, HDL-C, waist circumference (among men) and lower blood pressure. Vegetables/fruit dietary pattern did not protect against metabolic syndrome and newly diagnosed diabetes but was associated with lower waist circumference. The inverse associations found for the dietary pattern characterizing Brazilian fast foods and desserts, typically containing dairy products, with newly diagnosed diabetes, and for the DASH diet with metabolic syndrome, support previously demonstrated beneficial effects of dairy products in metabolism. The positive association with metabolic syndrome and newly diagnosed diabetes found for the pattern characterizing a typical Brazilian meal deserves further investigation, particularly since it is frequently accompanied by processed meat. Trial registration NCT02320461. Registered 18 December 2014.

Platelet-rich plasma supplementation in arthroscopic repair of full-thickness rotator cuff tears: a randomized clinical trial.

PubMed

D'Ambrosi, R; Palumbo, F; Paronzini, A; Ragone, V; Facchini, R M

2016-12-01

Results on the effectiveness of PRP supplementation in arthroscopic rotator cuff repair are conflicting, making it difficult to draw definitive conclusions. This was a prospective, randomized, and double-blind study with two groups of 20 patients each (PRP group and control group). Degenerative supraspinatus full-thickness tears grade C2-C3 were subjected to arthroscopic repair; PRP supplementation was given to patients in the PRP group. The outcomes were assessed by DASH, Constant scales, and ultrasound before and 6 months after surgery. Pain measured by VAS was evaluated preoperatively and 7 and 30 days after surgery. The two groups did not differ significantly by age, sex, and dominance of the affected side. In all surgical procedures, a long head of the biceps tenotomy and single-row repair were performed. The preoperative VAS was 5.6 ± 2.4 in PRP group and 6.4 ± 1.5 in the control group (p > 0.05). The group supplemented with PRP reported a VAS significantly better in the first week (2.5 ± 1.9 vs 5.3 ± 2.1, p < 0.05) and during the first month after surgery (1.5 ± 1.0 vs 3.2 ± 1.7, p < 0.05) compared to the control group. The preoperative Constant and DASH scores were 39.95 ± 12 and 51 ± 15.2, respectively, in the PRP group and 41 ± 11 (p > 0.05) and 45 ± 12.6 (p > 0.05) in the control group. The average Constant score improved significantly after 6 months to 81 ± 11.2 (p < 0.05) in the PRP group and 78.5 ± 9 (p < 0.05) in the control group. No differences were noted between the two groups (p > 0.05). The DASH score after 6 months was 17.4 ± 8 (p < 0.05) for the treatment group (the PRP group) and 21 ± 8.4 (p < 0.05) for the control group. No statistically significant differences were found as regards the DASH score in the two groups after 6 months (p > 0.05). The two groups showed no differences in the ultrasound evaluation after 6 months either. No re-ruptures occurred in either group. PRP leads to a reduction in pain during a short-term follow-up. Pain reduction allows for a more rapid recovery of mobilization and improvement in functionality. Randomized controlled trial, Level of evidence, 1.

Structural Mechanisms of Inactivation in Scabies Mite Serine Protease Paralogues

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fischer, Katja; Langendorf, Christopher G.; Irving, James A.

2009-08-07

The scabies mite (Sarcoptes scabiei) is a parasite responsible for major morbidity in disadvantaged communities and immuno-compromised patients worldwide. In addition to the physical discomfort caused by the disease, scabies infestations facilitate infection by Streptococcal species via skin lesions, resulting in a high prevalence of rheumatic fever/heart disease in affected communities. The scabies mite produces 33 proteins that are closely related to those in the dust mite group 3 allergen and belong to the S1-like protease family (chymotrypsin-like). However, all but one of these molecules contain mutations in the conserved active-site catalytic triad that are predicted to render them catalyticallymore » inactive. These molecules are thus termed scabies mite inactivated protease paralogues (SMIPPs). The precise function of SMIPPs is unclear; however, it has been suggested that these proteins might function by binding and protecting target substrates from cleavage by host immune proteases, thus preventing the host from mounting an effective immune challenge. In order to begin to understand the structural basis for SMIPP function, we solved the crystal structures of SMIPP-S-I1 and SMIPP-S-D1 at 1.85 {angstrom} and 2.0 {angstrom} resolution, respectively. Both structures adopt the characteristic serine protease fold, albeit with large structural variations over much of the molecule. In both structures, mutations in the catalytic triad together with occlusion of the S1 subsite by a conserved Tyr200 residue is predicted to block substrate ingress. Accordingly, we show that both proteases lack catalytic function. Attempts to restore function (via site-directed mutagenesis of catalytic residues as well as Tyr200) were unsuccessful. Taken together, these data suggest that SMIPPs have lost the ability to bind substrates in a classical 'canonical' fashion, and instead have evolved alternative functions in the lifecycle of the scabies mite.« less

Sweet potato SPAP1 is a typical aspartic protease and participates in ethephon-mediated leaf senescence.

PubMed

Chen, Hsien-Jung; Huang, Yu-Hsuan; Huang, Guan-Jhong; Huang, Shyh-Shyun; Chow, Te-Jin; Lin, Yaw-Huei

2015-05-15

Plant aspartic proteases are generally divided into three categories: typical, nucellin-like, and atypical aspartic proteases based on their gene and protein structures. In this report, a full-length cDNA SPAP1 was cloned from sweet potato leaves, which contained 1515 nucleotides (504 amino acids) and exhibited high amino acid sequence identity (ca. 51-72%) with plant typical aspartic proteases, including tomato LeAspP, potato StAsp, and wheat WAP2. SPAP1 also contained conserved DTG and DSG amino acid residues within its catalytic domain and plant specific insert (PSI) at the C-terminus. The cDNA corresponding to the mature protein (starting from the 66th to 311th amino acid residues) without PSI domain was constructed with pET30a expression vector for fusion protein and antibody production. RT-PCR and protein blot hybridization showed that SPAP1 expression level was the highest in L3 mature leaves, then gradually declined until L5 completely yellow leaves. Ethephon, an ethylene-releasing compound, also enhanced SPAP1 expression at the time much earlier than the onset of leaf senescence. Exogenous application of SPAP1 fusion protein promoted ethephon-induced leaf senescence, which could be abolished by pre-treatment of SPAP1 fusion protein with (a) 95 °C for 5 min, (b) aspartic protease inhibitor pepstatin A, and (c) anti-SPAP1 antibody, respectively. Exogenous SPAP1 fusion protein, whereas, did not significantly affect leaf senescence under dark. These data conclude that sweet potato SPAP1 is a functional typical aspartic protease and participates in ethephon-mediated leaf senescence. The SPAP1-promoted leaf senescence and its activity are likely not associated with the PSI domain. Interaction of ethephon-inducible components for effective SPAP1 promotion on leaf senescence is also suggested. Copyright © 2015 Elsevier GmbH. All rights reserved.

Clinical outcomes after arthroscopic release for recalcitrant frozen shoulder.

PubMed

Ebrahimzadeh, Mohammad H; Moradi, Ali; Pour, Mostafa Khalili; Moghadam, Mohammad Hallaj; Kachooei, Amir Reza

2014-09-01

To explain the role of arthroscopic release in intractable frozen shoulders. We used different questionnaires and measuring tools to understand whether arthroscopic release is the superior modality to treat patients with intractable frozen shoulders. Between 2007 and 2013, in a prospective study, we enrolled 80 patients (52 females and 28 males) with recalcitrant frozen shoulder, who underwent arthroscopic release at Ghaem Hospital, a tertiary referral center, in Mashhad, Iran. Before operation, all patients filled out the Disability of Arm, Shoulder and Hand (DASH), Constant, University of California Los Angeles (UCLA), ROWE and Visual Analogue Scale (VAS) for pain questionnaires. We measured the difference in range of motion between both the normal and the frozen shoulders in each patient. The average age of the patients was 50.8±7.1 years. In 49 patients, the right shoulder was affected and in the remaining 31 the left side was affected. Before surgery, the patients were suffering from this disease on average for 11.7±10.3 months. The average time to follow-up was 47.2±6.8 months (14 to 60 months). Diabetes mellitus (38%) and history of shoulder trauma (23%) were the most common comorbidities in our patients. We did not find any significant differences between baseline characteristics of diabetics patients with non-diabetics ones. After surgery, the average time to achieve maximum pain improvement and range of motion were 3.6±2.1 and 3.6±2 months, respectively. The VAS score, constant shoulder score, Rowe score, UCLA shoulder score, and DASH score showed significant improvement in shoulder function after surgery, and shoulder range of motion improved in all directions compared to pre-operation range of motion. According to our results, arthroscopic release of recalcitrant frozen shoulder is a valuable modality in treating this disease. This method could decrease pain and improve both subjective and objective mid-term outcomes.

Clinical Outcomes after Arthroscopic Release for Recalcitrant Frozen Shoulder

PubMed Central

Ebrahimzadeh, Mohammad H; Moradi, Ali; Pour, Mostafa Khalili; Moghadam, Mohammad Hallaj; Kachooei, Amir Reza

2014-01-01

Background: To explain the role of arthroscopic release in intractable frozen shoulders. We used different questionnaires and measuring tools to understand whether arthroscopic release is the superior modality to treat patients with intractable frozen shoulders. Methods: Between 2007 and 2013, in a prospective study, we enrolled 80 patients (52 females and 28 males) with recalcitrant frozen shoulder, who underwent arthroscopic release at Ghaem Hospital, a tertiary referral center, in Mashhad, Iran. Before operation, all patients filled out the Disability of Arm, Shoulder and Hand (DASH), Constant, University of California Los Angeles (UCLA), ROWE and Visual Analogue Scale (VAS) for pain questionnaires. We measured the difference in range of motion between both the normal and the frozen shoulders in each patient. Results: The average age of the patients was 50.8±7.1 years. In 49 patients, the right shoulder was affected and in the remaining 31 the left side was affected. Before surgery, the patients were suffering from this disease on average for 11.7±10.3 months. The average time to follow-up was 47.2±6.8 months (14 to 60 months). Diabetes mellitus (38%) and history of shoulder trauma (23%) were the most common comorbidities in our patients. We did not find any significant differences between baseline characteristics of diabetics patients with non-diabetics ones. After surgery, the average time to achieve maximum pain improvement and range of motion were 3.6±2.1 and 3.6±2 months, respectively. The VAS score, constant shoulder score, Rowe score, UCLA shoulder score, and DASH score showed significant improvement in shoulder function after surgery, and shoulder range of motion improved in all directions compared to pre-operation range of motion. Conclusions: According to our results, arthroscopic release of recalcitrant frozen shoulder is a valuable modality in treating this disease. This method could decrease pain and improve both subjective and objective mid-term outcomes. PMID:25386586

In vitro antioxidant activity of rice protein affected by alkaline degree and gastrointestinal protease digestion.

PubMed

Liu, Ye; Wang, Zhengxuan; Li, Hui; Liang, Mingcai; Yang, Lin

2016-12-01

To elucidate whether and how alkali treatment, which is a common process for rice protein (RP) extraction, affects antioxidant activity of RP, the different degree of alkali (from 0.1% to 0.4% of NaOH) was used to extract RP (RP-1, RP-2, RP-3, RP-4). The antioxidant capacities of scavenging free radicals [2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid] diammonium salt, ABTS; 1,1-diphenyl-2-picrylhydrazyl, DPPH), chelating metals (iron, copper) and reducing power investigated in the hydrolysates of RPs (RP-1, RP-2, RP-3, RP-4) during in vitro pepsin-pancreatin digestion were effectively affected by alkali treatment. The present study demonstrated that the weakest antioxidant responses to ABTS radical-scavenging activity, DPPH radical-scavenging activity, iron chelating activity, copper chelating activity and reducing power were produced by RP-4 extracted by the highest alkali proportion (0.4% NaOH). The present study indicates that antioxidant capacity of RP could be more readily depressed by strict alkali degree and affected by gastrointestinal proteases. Results suggest that alkali extraction is a vital process to regulate the antioxidant activity of RP through modifying the compositions of amino acids, which are dependent on alkali magnitude. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Cleavage Entropy as Quantitative Measure of Protease Specificity

PubMed Central

Fuchs, Julian E.; von Grafenstein, Susanne; Huber, Roland G.; Margreiter, Michael A.; Spitzer, Gudrun M.; Wallnoefer, Hannes G.; Liedl, Klaus R.

2013-01-01

A purely information theory-guided approach to quantitatively characterize protease specificity is established. We calculate an entropy value for each protease subpocket based on sequences of cleaved substrates extracted from the MEROPS database. We compare our results with known subpocket specificity profiles for individual proteases and protease groups (e.g. serine proteases, metallo proteases) and reflect them quantitatively. Summation of subpocket-wise cleavage entropy contributions yields a measure for overall protease substrate specificity. This total cleavage entropy allows ranking of different proteases with respect to their specificity, separating unspecific digestive enzymes showing high total cleavage entropy from specific proteases involved in signaling cascades. The development of a quantitative cleavage entropy score allows an unbiased comparison of subpocket-wise and overall protease specificity. Thus, it enables assessment of relative importance of physicochemical and structural descriptors in protease recognition. We present an exemplary application of cleavage entropy in tracing substrate specificity in protease evolution. This highlights the wide range of substrate promiscuity within homologue proteases and hence the heavy impact of a limited number of mutations on individual substrate specificity. PMID:23637583

Subfamily-Specific Fluorescent Probes for Cysteine Proteases Display Dynamic Protease Activities during Seed Germination.

PubMed

Lu, Haibin; Chandrasekar, Balakumaran; Oeljeklaus, Julian; Misas-Villamil, Johana C; Wang, Zheming; Shindo, Takayuki; Bogyo, Matthew; Kaiser, Markus; van der Hoorn, Renier A L

2015-08-01

Cysteine proteases are an important class of enzymes implicated in both developmental and defense-related programmed cell death and other biological processes in plants. Because there are dozens of cysteine proteases that are posttranslationally regulated by processing, environmental conditions, and inhibitors, new methodologies are required to study these pivotal enzymes individually. Here, we introduce fluorescence activity-based probes that specifically target three distinct cysteine protease subfamilies: aleurain-like proteases, cathepsin B-like proteases, and vacuolar processing enzymes. We applied protease activity profiling with these new probes on Arabidopsis (Arabidopsis thaliana) protease knockout lines and agroinfiltrated leaves to identify the probe targets and on other plant species to demonstrate their broad applicability. These probes revealed that most commercially available protease inhibitors target unexpected proteases in plants. When applied on germinating seeds, these probes reveal dynamic activities of aleurain-like proteases, cathepsin B-like proteases, and vacuolar processing enzymes, coinciding with the remobilization of seed storage proteins. © 2015 American Society of Plant Biologists. All Rights Reserved.

Diversity of Both the Cultivable Protease-Producing Bacteria and Bacterial Extracellular Proteases in the Coastal Sediments of King George Island, Antarctica

PubMed Central

Zhou, Ming-Yang; Wang, Guang-Long; Li, Dan; Zhao, Dian-Li; Qin, Qi-Long; Chen, Xiu-Lan; Chen, Bo; Zhou, Bai-Cheng; Zhang, Xi-Ying; Zhang, Yu-Zhong

2013-01-01

Protease-producing bacteria play a vital role in degrading sedimentary organic nitrogen. However, the diversity of these bacteria and their extracellular proteases in most regions remain unknown. In this paper, the diversity of the cultivable protease-producing bacteria and of bacterial extracellular proteases in the sediments of Maxwell Bay, King George Island, Antarctica was investigated. The cultivable protease-producing bacteria reached 105 cells/g in all 8 sediment samples. The cultivated protease-producing bacteria were mainly affiliated with the phyla Actinobacteria, Firmicutes, Bacteroidetes, and Proteobacteria, and the predominant genera were Bacillus (22.9%), Flavobacterium (21.0%) and Lacinutrix (16.2%). Among these strains, Pseudoalteromonas and Flavobacteria showed relatively high protease production. Inhibitor analysis showed that nearly all the extracellular proteases from the bacteria were serine proteases or metalloproteases. These results begin to address the diversity of protease-producing bacteria and bacterial extracellular proteases in the sediments of the Antarctic Sea. PMID:24223990

Antimicrobial activity of an aspartic protease from Salpichroa origanifolia fruits.

PubMed

Díaz, M E; Rocha, G F; Kise, F; Rosso, A M; Guevara, M G; Parisi, M G

2018-05-08

Plant proteases play a fundamental role in several processes like growth, development and in response to biotic and abiotic stress. In particular, aspartic proteases (AP) are expressed in different plant organs and have antimicrobial activity. Previously, we purified an AP from Salpichroa origanifolia fruits called salpichroin. The aim of this work was to determine the cytotoxic activity of this enzyme on selected plant and human pathogens. For this purpose, the growth of the selected pathogens was analysed after exposure to different concentrations of salpichroin. The results showed that the enzyme was capable of inhibiting Fusarium solani and Staphylococcus aureus in a dose-dependent manner. It was determined that 1·2 μmol l -1 of salpichroin was necessary to inhibit 50% of conidial germination, and the minimal bactericidal concentration was between 1·9 and 2·5 μmol l -1 . Using SYTOX Green dye we were able to demonstrate that salpichroin cause membrane permeabilization. Moreover, the enzyme treated with its specific inhibitor pepstatin A did not lose its antibacterial activity. This finding demonstrates that the cytotoxic activity of salpichroin is due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of the AP could represent a potential alternative for the control of pathogens that affect humans or crops of economic interest. This study provides insights into the antimicrobial activity of an aspartic protease isolated from Salpichroa origanifolia fruits on plant and human pathogens. The proteinase inhibited Fusarium solani and Staphylococcus aureus in a dose-dependent manner due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of salpichroin suggests its potential applications as an important tool for the control of pathogenic micro-organisms affecting humans and crops of economic interest. Therefore, it would represent a new alternative to avoid the problems of environmental pollution and antimicrobial resistance. © 2018 The Society for Applied Microbiology.

Loss of Drosophila i-AAA protease, dYME1L, causes abnormal mitochondria and apoptotic degeneration.

PubMed

Qi, Y; Liu, H; Daniels, M P; Zhang, G; Xu, H

2016-02-01

Mitochondrial AAA (ATPases Associated with diverse cellular Activities) proteases i-AAA (intermembrane space-AAA) and m-AAA (matrix-AAA) are closely related and have major roles in inner membrane protein homeostasis. Mutations of m-AAA proteases are associated with neuromuscular disorders in humans. However, the role of i-AAA in metazoans is poorly understood. We generated a deletion affecting Drosophila i-AAA, dYME1L (dYME1L(del)). Mutant flies exhibited premature aging, progressive locomotor deficiency and neurodegeneration that resemble some key features of m-AAA diseases. dYME1L(del) flies displayed elevated mitochondrial unfolded protein stress and irregular cristae. Aged dYME1L(del) flies had reduced complex I (NADH/ubiquinone oxidoreductase) activity, increased level of reactive oxygen species (ROS), severely disorganized mitochondrial membranes and increased apoptosis. Furthermore, inhibiting apoptosis by targeting dOmi (Drosophila Htra2/Omi) or DIAP1, or reducing ROS accumulation suppressed retinal degeneration. Our results suggest that i-AAA is essential for removing unfolded proteins and maintaining mitochondrial membrane architecture. Loss of i-AAA leads to the accumulation of oxidative damage and progressive deterioration of membrane integrity, which might contribute to apoptosis upon the release of proapoptotic molecules such as dOmi. Containing ROS level could be a potential strategy to manage mitochondrial AAA protease deficiency.

Activities of Vacuolar Cysteine Proteases in Plant Senescence.

PubMed

Martínez, Dana E; Costa, Lorenza; Guiamét, Juan José

2018-01-01

Plant senescence is accompanied by a marked increase in proteolytic activities, and cysteine proteases (Cys-protease) represent the prevailing class among the responsible proteases. Cys-proteases predominantly locate to lytic compartments, i.e., to the central vacuole (CV) and to senescence-associated vacuoles (SAVs), the latter being specific to the photosynthetic cells of senescing leaves. Cellular fractionation of vacuolar compartments may facilitate Cys-proteases purification and their concentration for further analysis. Active Cys-proteases may be analyzed by different, albeit complementary approaches: (1) in vivo examination of proteolytic activity by fluorescence microscopy using specific substrates which become fluorescent upon cleavage by Cys-proteases, (2) protease labeling with specific probes that react irreversibly with the active enzymes, and (3) zymography, whereby protease activities are detected in polyacrylamide gels copolymerized with a substrate for proteases. Here we describe the three methods mentioned above for detection of active Cys-proteases and a cellular fractionation technique to isolate SAVs.

Interaction between FMDV Lpro and transcription factor ADNP is required for viral replication

USDA-ARS?s Scientific Manuscript database

The foot-and-mouth disease virus (FMDV) leader protease (Lpro) inhibits host translation and transcription affecting the expression of several factors involved in innate immunity. In this study, we have identified the host transcription factor ADNP (activity dependent neuroprotective protein) as an ...

Limited amplification of chronic wasting disease prions in the peripheral tissues of intracerebrally inoculated cattle

USDA-ARS?s Scientific Manuscript database

Chronic wasting disease (CWD) is a fatal neurodegenerative disease, classified as a prion disease or transmissible spongiform encephalopathy (TSE) similar to bovine spongiform encephalopathy (BSE). Cervids affected by CWD accumulate an abnormal protease resistant prion protein throughout the central...

Cathepsin B Activity Initiates Apoptosis via Digestive Protease Activation in Pancreatic Acinar Cells and Experimental Pancreatitis*

PubMed Central

Sendler, Matthias; Maertin, Sandrina; John, Daniel; Persike, Maria; Weiss, F. Ulrich; Krüger, Burkhard; Wartmann, Thomas; Wagh, Preshit; Halangk, Walter; Schaschke, Norbert; Mayerle, Julia; Lerch, Markus M.

2016-01-01

Pancreatitis is associated with premature activation of digestive proteases in the pancreas. The lysosomal hydrolase cathepsin B (CTSB) is a known activator of trypsinogen, and its deletion reduces disease severity in experimental pancreatitis. Here we studied the activation mechanism and subcellular compartment in which CTSB regulates protease activation and cellular injury. Cholecystokinin (CCK) increased the activity of CTSB, cathepsin L, trypsin, chymotrypsin, and caspase 3 in vivo and in vitro and induced redistribution of CTSB to a secretory vesicle-enriched fraction. Neither CTSB protein nor activity redistributed to the cytosol, where the CTSB inhibitors cystatin-B/C were abundantly present. Deletion of CTSB reduced and deletion of cathepsin L increased intracellular trypsin activation. CTSB deletion also abolished CCK-induced caspase 3 activation, apoptosis-inducing factor, as well as X-linked inhibitor of apoptosis protein degradation, but these depended on trypsinogen activation via CTSB. Raising the vesicular pH, but not trypsin inhibition, reduced CTSB activity. Trypsin inhibition did not affect apoptosis in hepatocytes. Deletion of CTSB affected apoptotic but not necrotic acinar cell death. In summary, CTSB in pancreatitis undergoes activation in a secretory, vesicular, and acidic compartment where it activates trypsinogen. Its deletion or inhibition regulates acinar cell apoptosis but not necrosis in two models of pancreatitis. Caspase 3-mediated apoptosis depends on intravesicular trypsinogen activation induced by CTSB, not CTSB activity directly, and this mechanism is pancreas-specific. PMID:27226576

Cathepsin B Activity Initiates Apoptosis via Digestive Protease Activation in Pancreatic Acinar Cells and Experimental Pancreatitis.

PubMed

Sendler, Matthias; Maertin, Sandrina; John, Daniel; Persike, Maria; Weiss, F Ulrich; Krüger, Burkhard; Wartmann, Thomas; Wagh, Preshit; Halangk, Walter; Schaschke, Norbert; Mayerle, Julia; Lerch, Markus M

2016-07-08

Pancreatitis is associated with premature activation of digestive proteases in the pancreas. The lysosomal hydrolase cathepsin B (CTSB) is a known activator of trypsinogen, and its deletion reduces disease severity in experimental pancreatitis. Here we studied the activation mechanism and subcellular compartment in which CTSB regulates protease activation and cellular injury. Cholecystokinin (CCK) increased the activity of CTSB, cathepsin L, trypsin, chymotrypsin, and caspase 3 in vivo and in vitro and induced redistribution of CTSB to a secretory vesicle-enriched fraction. Neither CTSB protein nor activity redistributed to the cytosol, where the CTSB inhibitors cystatin-B/C were abundantly present. Deletion of CTSB reduced and deletion of cathepsin L increased intracellular trypsin activation. CTSB deletion also abolished CCK-induced caspase 3 activation, apoptosis-inducing factor, as well as X-linked inhibitor of apoptosis protein degradation, but these depended on trypsinogen activation via CTSB. Raising the vesicular pH, but not trypsin inhibition, reduced CTSB activity. Trypsin inhibition did not affect apoptosis in hepatocytes. Deletion of CTSB affected apoptotic but not necrotic acinar cell death. In summary, CTSB in pancreatitis undergoes activation in a secretory, vesicular, and acidic compartment where it activates trypsinogen. Its deletion or inhibition regulates acinar cell apoptosis but not necrosis in two models of pancreatitis. Caspase 3-mediated apoptosis depends on intravesicular trypsinogen activation induced by CTSB, not CTSB activity directly, and this mechanism is pancreas-specific. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

The effect of different protease inhibitors on stability of parathyroid hormone, insulin, and prolactin levels under different lag times and storage conditions until analysis.

PubMed

Baykan, Ozgur; Yaman, Ali; Gerin, Fethullah; Sirikci, Onder; Haklar, Goncagul

2017-11-01

Proteolytic cleavage through proteases affects peptide hormone levels, which is of particular significance when the time interval between sampling and analysis is prolonged. We evaluated the stability of parathyroid hormone, insulin, and prolactin molecules (i) with different protease inhibitors such as K 2 EDTA, aprotinin, and protease inhibitor cocktail (PIC), (ii) with different lag times (6-72 hours), and (iii) under different storage temperatures (4°C vs room temperature [RT]) until analysis. Blood samples were collected into 2 sets of 5 Vacutainer ® tubes (Becton Dickinson) from 10 healthy adults. Tubes 1 and 2 were plain gel separator tubes. Tubes 3, 4, and 5 contained PIC (1%), aprotinin (500 KIU/mL), and K 2 EDTA, respectively. After centrifugation at 1300 g for 10 minutes, PIC added to tube 2 of each set. Samples were analyzed and then one set was stored at 4°C, whereas the other at RT until analysis at 6, 24, 48, and 72 hours. Hormone levels were determined with electrochemiluminescence immunoassay (ModularE170; Roche Diagnostics). The results were compared with desirable bias limits (DBL) from Westgard QC database. Insulin at RT decreases exceeding the DBL starting from 24 hours and K 2 EDTA preserved insulin. PTH exceeded the DBL at RT for 48 hours or longer and PIC addition after centrifugation inhibited its degradation. Prolactin remained stable in all tested conditions. All parameters in the plain gel separator tubes remained within DBL when stored at 4°C until 72 hours. Different proteases may degrade peptide hormones and measures should be taken to counteract these effects especially if there is a delay before analysis. © 2017 Wiley Periodicals, Inc.

A Potential Mechanism for ADC-Induced Neutropenia: Role of Neutrophils in Their Own Demise.

PubMed

Zhao, Hui; Gulesserian, Sara; Malinao, Maria Christina; Ganesan, Sathish Kumar; Song, James; Chang, Mi Sook; Williams, Melissa M; Zeng, Zhilan; Mattie, Michael; Mendelsohn, Brian A; Stover, David R; Doñate, Fernando

2017-09-01

Neutropenia is a common adverse event in cancer patients treated with antibody-drug conjugates (ADC) and we aimed to elucidate the potential mechanism of this toxicity. To investigate whether ADCs affect neutrophil production from bone marrow, an in vitro assay was developed in which hematopoietic stem cells (HSC) were differentiated to neutrophils. Several antibodies against targets absent in HSCs and neutrophils were conjugated to MMAE via a cleavable valine-citrulline linker (vcMMAE-ADC) or MMAF via a noncleavable maleimidocaproyl linker (mcMMAF-ADC), and their cytotoxicity was tested in the neutrophil differentiation assay. Results showed that HSCs had similar sensitivity to vcMMAE-ADCs and mcMMAF-ADCs; however, vcMMAE-ADCs were more cytotoxic to differentiating neutrophils than the same antibody conjugated to mcMMAF. This inhibitory effect was not mediated by internalization of ADC either by macropinocytosis or FcγRs. Our results suggested that extracellular proteolysis of the cleavable valine-citrulline linker is responsible for the cytotoxicity to differentiating neutrophils. Mass spectrometry analyses indicated that free MMAE was released from vcMMAE-ADCs in the extracellular compartment when they were incubated with differentiating neutrophils or neutrophil conditioned medium, but not with HSC-conditioned medium. Using different protease inhibitors, our data suggested that serine, but not cysteine proteases, were responsible for the cleavage. In vitro experiments demonstrated that the purified serine protease, elastase, was capable of releasing free MMAE from a vcMMAE-ADC. Here we propose that ADCs containing protease cleavable linkers can contribute to neutropenia via extracellular cleavage mediated by serine proteases secreted by differentiating neutrophils in bone marrow. Mol Cancer Ther; 16(9); 1866-76. ©2017 AACR See related article by Zhao et al., p. 1877 . ©2017 American Association for Cancer Research.

Indigenous proteases in the skin of unicorn leatherjacket (Alutherus monoceros) and their influence on characteristic and functional properties of gelatin.

PubMed

Ahmad, Mehraj; Benjakul, Soottawat; Ovissipour, Mahmoudreza; Prodpran, Thummanoon

2011-07-15

Indigenous proteases in the skin of unicorn leatherjacket (Alutherus monoceros) were characterised using autolytic study. Maximised autolysis was found at pH 7 and 50°C. Autolysis was markedly inhibited by 0.04mM soybean trypsin inhibitor (SBTI), suggesting that heat activated serine protease was predominant in the skin. The impact of indigenous proteases on the properties of gelatin extracted from unicorn leatherjacket skin was investigated. Gelatin was extracted from unicorn leatherjacket skin using distilled water at 50°C for 12h in the presence and absence of 0.04mM SBTI. In the presence of SBTI, the degradation was markedly inhibited, but a lower gelatin extraction yield was obtained (P<0.05). Extracted gelatins contained α(1) and α(2) chains as the predominant components with some degradation peptides. FTIR spectra indicated a greater loss of molecular order of the triple helix and a higher degradation was found in gelatin extracted in the absence of 0.04mM SBTI. The net charge of gelatin samples extracted with and without 0.04mM SBTI became zero at pHs of 8.45 and 7.31, respectively, as determined by ζ-potential titration. Higher gel strength (320.68±3.02g) was obtained in gelatin extracted with SBTI, compared with that of gelatin extracted without SBTI (288.63±1.44g). High emulsifying activity index but lower emulsifying stability index was observed in the former. Therefore, heat-activated serine protease was involved in the degradation of gelatin molecules, thereby affecting the yield, proteinaceous components and properties of gelatin from unicorn leatherjacket skin. Copyright © 2011 Elsevier Ltd. All rights reserved.

Recombinant Passenger Proteins Can Be Conveniently Purified by One-Step Affinity Chromatography.

PubMed

Wang, Hua-zhen; Chu, Zhi-zhan; Chen, Chang-chao; Cao, Ao-cheng; Tong, Xin; Ouyang, Can-bin; Yuan, Qi-hang; Wang, Mi-nan; Wu, Zhong-kun; Wang, Hai-hong; Wang, Sheng-bin

2015-01-01

Fusion tag is one of the best available tools to date for enhancement of the solubility or improvement of the expression level of recombinant proteins in Escherichia coli. Typically, two consecutive affinity purification steps are often necessitated for the purification of passenger proteins. As a fusion tag, acyl carrier protein (ACP) could greatly increase the soluble expression level of Glucokinase (GlcK), α-Amylase (Amy) and GFP. When fusion protein ACP-G2-GlcK-Histag and ACP-G2-Amy-Histag, in which a protease TEV recognition site was inserted between the fusion tag and passenger protein, were coexpressed with protease TEV respectively in E. coli, the efficient intracellular processing of fusion proteins was achieved. The resulting passenger protein GlcK-Histag and Amy-Histag accumulated predominantly in a soluble form, and could be conveniently purified by one-step Ni-chelating chromatography. However, the fusion protein ACP-GFP-Histag was processed incompletely by the protease TEV coexpressed in vivo, and a large portion of the resulting target protein GFP-Histag aggregated in insoluble form, indicating that the intracellular processing may affect the solubility of cleaved passenger protein. In this context, the soluble fusion protein ACP-GFP-Histag, contained in the supernatant of E. coli cell lysate, was directly subjected to cleavage in vitro by mixing it with the clarified cell lysate of E. coli overexpressing protease TEV. Consequently, the resulting target protein GFP-Histag could accumulate predominantly in a soluble form, and be purified conveniently by one-step Ni-chelating chromatography. The approaches presented here greatly simplify the purification process of passenger proteins, and eliminate the use of large amounts of pure site-specific proteases.

Recombinant Passenger Proteins Can Be Conveniently Purified by One-Step Affinity Chromatography

PubMed Central

Wang, Hua-zhen; Chu, Zhi-zhan; Chen, Chang-chao; Cao, Ao-cheng; Tong, Xin; Ouyang, Can-bin; Yuan, Qi-hang; Wang, Mi-nan; Wu, Zhong-kun; Wang, Hai-hong; Wang, Sheng-bin

2015-01-01

Fusion tag is one of the best available tools to date for enhancement of the solubility or improvement of the expression level of recombinant proteins in Escherichia coli. Typically, two consecutive affinity purification steps are often necessitated for the purification of passenger proteins. As a fusion tag, acyl carrier protein (ACP) could greatly increase the soluble expression level of Glucokinase (GlcK), α-Amylase (Amy) and GFP. When fusion protein ACP-G2-GlcK-Histag and ACP-G2-Amy-Histag, in which a protease TEV recognition site was inserted between the fusion tag and passenger protein, were coexpressed with protease TEV respectively in E. coli, the efficient intracellular processing of fusion proteins was achieved. The resulting passenger protein GlcK-Histag and Amy-Histag accumulated predominantly in a soluble form, and could be conveniently purified by one-step Ni-chelating chromatography. However, the fusion protein ACP-GFP-Histag was processed incompletely by the protease TEV coexpressed in vivo, and a large portion of the resulting target protein GFP-Histag aggregated in insoluble form, indicating that the intracellular processing may affect the solubility of cleaved passenger protein. In this context, the soluble fusion protein ACP-GFP-Histag, contained in the supernatant of E. coli cell lysate, was directly subjected to cleavage in vitro by mixing it with the clarified cell lysate of E. coli overexpressing protease TEV. Consequently, the resulting target protein GFP-Histag could accumulate predominantly in a soluble form, and be purified conveniently by one-step Ni-chelating chromatography. The approaches presented here greatly simplify the purification process of passenger proteins, and eliminate the use of large amounts of pure site-specific proteases. PMID:26641240

Inhibition of protein carbamylation in urea solution using ammonium-containing buffers.

PubMed

Sun, Shisheng; Zhou, Jian-Ying; Yang, Weiming; Zhang, Hui

2014-02-01

Urea solution is one of the most commonly employed protein denaturants for protease digestion in proteomic studies. However, it has long been recognized that urea solution can cause carbamylation at the N termini of proteins/peptides and at the side chain amino groups of lysine and arginine residues. Protein/peptide carbamylation blocks protease digestion and affects protein identification and quantification in mass spectrometry analysis by blocking peptide amino groups from isotopic/isobaric labeling and changing peptide charge states, retention times, and masses. In addition, protein carbamylation during sample preparation makes it difficult to study in vivo protein carbamylation. In this study, we compared the peptide carbamylation in urea solutions of different buffers and found that ammonium-containing buffers were the most effective buffers to inhibit protein carbamylation in urea solution. The possible mechanism of carbamylation inhibition by ammonium-containing buffers is discussed, and a revised procedure for the protease digestion of proteins in urea and ammonium-containing buffers was developed to facilitate its application in proteomic research. Copyright © 2013 Elsevier Inc. All rights reserved.

An integrated proteomic approach uncovers novel substrates and functions of the Lon protease in Escherichia coli.

PubMed

Arends, Jan; Griego, Marcena; Thomanek, Nikolas; Lindemann, Claudia; Kutscher, Blanka; Meyer, Helmut E; Narberhaus, Franz

2018-04-30

Controlling the cellular abundance and proper function of proteins by proteolysis is a universal process in all living organisms. In Escherichia coli, the ATP-dependent Lon protease is crucial for protein quality control and regulatory processes. To understand how diverse substrates are selected and degraded, unbiased global approaches are needed. We employed a quantitative Super-SILAC mass spectrometry approach and compared the proteomes of a lon mutant and a strain producing the protease to discover Lon-dependent physiological functions. To identify Lon substrates, we took advantage of a Lon trapping variant, which is able to translocate substrates but unable to degrade them. Lon-associated proteins were identified by label-free LC-MS/MS. The combination of both approaches revealed a total of 14 novel Lon substrates. Besides the identification of known pathways affected by Lon, for example the superoxide-stress response, our cumulative data suggests previously unrecognized fundamental functions of Lon in sulfur assimilation, nucleotide biosynthesis, amino acid and central energy metabolism. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Cathepsins: Proteases that are vital for survival but can also be fatal.

PubMed

Patel, Seema; Homaei, Ahmad; El-Seedi, Hesham R; Akhtar, Nadeem

2018-06-06

The state of enzymes in the human body determines the normal physiology or pathology, so all the six classes of enzymes are crucial. Proteases, the hydrolases, can be of several types based on the nucleophilic amino acid or the metal cofactor needed for their activity. Cathepsins are proteases with serine, cysteine, or aspartic acid residues as the nucleophiles, which are vital for digestion, coagulation, immune response, adipogenesis, hormone liberation, peptide synthesis, among a litany of other functions. But inflammatory state radically affects their normal roles. Released from the lysosomes, they degrade extracellular matrix proteins such as collagen and elastin, mediating parasite infection, autoimmune diseases, tumor metastasis, cardiovascular issues, and neural degeneration, among other health hazards. Over the years, the different types and isoforms of cathepsin, their optimal pH and functions have been studied, yet much information is still elusive. By taming and harnessing cathepsins, by inhibitors and judicious lifestyle, a gamut of malignancies can be resolved. This review discusses these aspects, which can be of clinical relevance. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

HtrA Is Important for Stress Resistance and Virulence in Haemophilus parasuis

PubMed Central

Zhang, Luhua; Li, Ying; Wen, Yiping; Lau, Gee W.; Huang, Xiaobo; Wu, Rui; Yan, Qigui; Huang, Yong; Zhao, Qin; Ma, Xiaoping

2016-01-01

Haemophilus parasuis is an opportunistic pathogen that causes Glässer's disease in swine, with polyserositis, meningitis, and arthritis. The high-temperature requirement A (HtrA)-like protease, which is involved in protein quality control, has been reported to be a virulence factor in many pathogens. In this study, we showed that HtrA of H. parasuis (HpHtrA) exhibited both chaperone and protease activities. Finally, nickel import ATP-binding protein (NikE), periplasmic dipeptide transport protein (DppA), and outer membrane protein A (OmpA) were identified as proteolytic substrates for HpHtrA. The protease activity reached its maximum at 40°C in a time-dependent manner. Disruption of the htrA gene from strain SC1401 affected tolerance to temperature stress and resistance to complement-mediated killing. Furthermore, increased autoagglutination and biofilm formation were detected in the htrA mutant. In addition, the htrA mutant was significantly attenuated in virulence in the murine model of infection. Together, these data demonstrate that HpHtrA plays an important role in the virulence of H. parasuis. PMID:27217419

HtrA Is Important for Stress Resistance and Virulence in Haemophilus parasuis.

PubMed

Zhang, Luhua; Li, Ying; Wen, Yiping; Lau, Gee W; Huang, Xiaobo; Wu, Rui; Yan, Qigui; Huang, Yong; Zhao, Qin; Ma, Xiaoping; Wen, Xintian; Cao, Sanjie

2016-08-01

Haemophilus parasuis is an opportunistic pathogen that causes Glässer's disease in swine, with polyserositis, meningitis, and arthritis. The high-temperature requirement A (HtrA)-like protease, which is involved in protein quality control, has been reported to be a virulence factor in many pathogens. In this study, we showed that HtrA of H. parasuis (HpHtrA) exhibited both chaperone and protease activities. Finally, nickel import ATP-binding protein (NikE), periplasmic dipeptide transport protein (DppA), and outer membrane protein A (OmpA) were identified as proteolytic substrates for HpHtrA. The protease activity reached its maximum at 40°C in a time-dependent manner. Disruption of the htrA gene from strain SC1401 affected tolerance to temperature stress and resistance to complement-mediated killing. Furthermore, increased autoagglutination and biofilm formation were detected in the htrA mutant. In addition, the htrA mutant was significantly attenuated in virulence in the murine model of infection. Together, these data demonstrate that HpHtrA plays an important role in the virulence of H. parasuis. Copyright © 2016 Zhang et al.

Preliminary crystallographic analysis of avian infectious bronchitis virus main protease

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Jun; Shen, Wei; Liao, Ming, E-mail: mliao@scau.edu.cn

The avian infectious bronchitis virus main protease has been crystallized; crystals diffract to 2.7 Å resolution. Infectious bronchitis virus (IBV) is the prototype of the genus Coronavirus. It causes a highly contagious disease which affects the respiratory, reproductive, neurological and renal systems of chickens, resulting great economic losses in the poultry industry worldwide. The coronavirus (CoV) main protease (M{sup pro}), which plays a pivotal role in viral gene expression and replication through a highly complex cascade involving the proteolytic processing of replicase polyproteins, is an attractive target for antiviral drug design. In this study, IBV M{sup pro} was overexpressed inmore » Escherichia coli. Crystals suitable for X-ray crystallography have been obtained using microseeding techniques and belong to space group P6{sub 1}22. X-ray diffraction data were collected in-house to 2.7 Å resolution from a single crystal. The unit-cell parameters were a = b = 119.1, c = 270.7 Å, α = β = 90, γ = 120°. Three molecules were predicted to be present in the asymmetric unit from a calculated self-rotation function.« less

Binding of Substrate Locks the Electrochemistry of CRY-DASH into DNA Repair.

PubMed

Gindt, Yvonne M; Messyasz, Adriana; Jumbo, Pamela I

2015-05-12

VcCry1, a member of the CRY-DASH family, may serve two diverse roles in vivo, including blue-light signaling and repair of UV-damaged DNA. We have discovered that the electrochemistry of the flavin adenine dinucleotide cofactor of VcCry1 is locked to cycle only between the hydroquinone and neutral semiquinone states when UV-damaged DNA is present. Other potential substrates, including undamaged DNA and ATP, have no discernible effect on the electrochemistry, and the kinetics of the reduction is unaffected by damaged DNA. Binding of the damaged DNA substrate determines the role of the protein and prevents the presumed photochemistry required for blue-light signaling.

Software Mechanisms for Multiprocessor TLB Consistency

DTIC Science & Technology

1989-12-01

thanks. Raj V aswani implemented the DASH message-passing system. Ramesh Govindan implemented part of the DASH virtual memory system. G . Scott...1 Latency (ma) 5𔃺 ---·-r··-·r···r··-T·-·-r··-·r····r····-r··-·r··- . g ~~~R=r 18 -----1·--·-r··-T·----1·--··r·---1·----1·--··r·---T·----1 16...model development. Synchronizing TLBs is similar to updating replicated data in a distributed environment. Lee and Garcia-Molina both used an M/ G /1

Plant cysteine proteases that evoke itch activate protease-activated receptors

PubMed Central

Reddy, V.B.; Lerner, E.A.

2013-01-01

Background Bromelain, ficin and papain are cysteine proteases from plants that produce itch upon injection into skin. Their mechanism of action has not been considered previously. Objectives To determine the mechanism by which these proteases function. Methods The ability of these proteases to activate protease-activated receptors was determined by ratiometric calcium imaging. Results We show here that bromelain, ficin and papain activate protease-activated receptors 2 and 4. Conclusions Bromelain, ficin and papain function as signalling molecules and activate protease-activated receptors. Activation of these receptors is the likely mechanism by which these proteases evoke itch. PMID:20491769

Characterization and identification of proteases secreted by Aspergillus fumigatus using free flow electrophoresis and MS.

PubMed

Neustadt, Madlen; Costina, Victor; Kupfahl, Claudio; Buchheidt, Dieter; Eckerskorn, Christoph; Neumaier, Michael; Findeisen, Peter

2009-06-01

Early diagnosis of life-threatening invasive aspergillosis in neutropenic patients remains challenging because current laboratory methods have limited diagnostic sensitivity and/or specificity. Aspergillus species are known to secrete various pathogenetically relevant proteases and the monitoring of their protease activity in serum specimens might serve as a new diagnostic approach.For the characterization and identification of secreted proteases, the culture supernatant of Aspergillus fumigatus was fractionated using free flow electrophoresis (Becton Dickinson). Protease activity of separated fractions was measured using fluorescently labeled reporter peptides. Fractions were also co-incubated in parallel with various protease inhibitors that specifically inhibit a distinct class of proteases e.g. metallo- or cysteine-proteases. Those fractions with high protease activity were further subjected to LC-MS/MS analysis for protease identification. The highest protease activity was measured in fractions with an acidic pH range. The results of the 'inhibitor-panel' gave a clear indication that it is mainly metallo- and serine-proteases that are involved in the degradation of reporter peptides. Furthermore, several proteases were identified that facilitate the optimization of reporter peptides for functional protease profiling as a diagnostic tool for invasive aspergillosis.

Active Site Characterization of Proteases Sequences from Different Species of Aspergillus.

PubMed

Morya, V K; Yadav, Virendra K; Yadav, Sangeeta; Yadav, Dinesh

2016-09-01

A total of 129 proteases sequences comprising 43 serine proteases, 36 aspartic proteases, 24 cysteine protease, 21 metalloproteases, and 05 neutral proteases from different Aspergillus species were analyzed for the catalytically active site residues using MEROPS database and various bioinformatics tools. Different proteases have predominance of variable active site residues. In case of 24 cysteine proteases of Aspergilli, the predominant active site residues observed were Gln193, Cys199, His364, Asn384 while for 43 serine proteases, the active site residues namely Asp164, His193, Asn284, Ser349 and Asp325, His357, Asn454, Ser519 were frequently observed. The analysis of 21 metalloproteases of Aspergilli revealed Glu298 and Glu388, Tyr476 as predominant active site residues. In general, Aspergilli species-specific active site residues were observed for different types of protease sequences analyzed. The phylogenetic analysis of these 129 proteases sequences revealed 14 different clans representing different types of proteases with diverse active site residues.

Dual origin of gut proteases in Formosan subterranean termites (Coptotermes formosanus Shiraki) (Isoptera: Rhinotermitidae).

PubMed

Sethi, Amit; Xue, Qing-Gang; La Peyre, Jerome F; Delatte, Jennifer; Husseneder, Claudia

2011-07-01

Cellulose digestion in lower termites, mediated by carbohydrases originating from both termite and endosymbionts, is well characterized. In contrast, limited information exists on gut proteases of lower termites, their origins and roles in termite nutrition. The objective of this study was to characterize gut proteases of the Formosan subterranean termite (Coptotermes formosanus Shiraki) (Isoptera: Rhinotermitidae). The protease activity of extracts from gut tissues (fore-, mid- and hindgut) and protozoa isolated from hindguts of termite workers was quantified using hide powder azure as a substrate and further characterized by zymography with gelatin SDS-PAGE. Midgut extracts showed the highest protease activity followed by the protozoa extracts. High level of protease activity was also detected in protozoa culture supernatants after 24 h incubation. Incubation of gut and protozoa extracts with class-specific protease inhibitors revealed that most of the proteases were serine proteases. All proteolytic bands identified after gelatin SDS-PAGE were also inhibited by serine protease inhibitors. Finally, incubation with chromogenic substrates indicated that extracts from fore- and hindgut tissues possessed proteases with almost exclusively trypsin-like activity while both midgut and protozoa extracts possessed proteases with trypsin-like and subtilisin/chymotrypsin-like activities. However, protozoa proteases were distinct from midgut proteases (with different molecular mass). Our results suggest that the Formosan subterranean termite not only produces endogenous proteases in its gut tissues, but also possesses proteases originating from its protozoan symbionts. Copyright © 2011 Elsevier Inc. All rights reserved.

Infrared study of matrix-isolated ethyl cyanide: simulation of the photochemistry in the atmosphere of Titan.

PubMed

Toumi, A; Piétri, N; Couturier-Tamburelli, I

2015-11-11

Low-temperature Ar matrix isolation has been carried out to investigate the infrared spectrum of ethyl cyanide (CH3CH2CN), a molecule present in the atmosphere of Titan. The λ > 120 nm and λ > 230 nm photolysis reactions of ethyl cyanide in an Ar matrix were also performed in order to compare the behaviour of this compound when it is submitted to high and low energetic radiations. These different wavelengths have been used with the aim to reproduce the radiation reaching the various parts of the atmosphere. Several photoproducts have been identified during photolysis such as vinyl cyanide (CH2[double bond, length as m-dash]CHCN), cyanoacetylene (HC3N), and ethylene/hydrogen cyanide (C2H4/HCN), ethylene/hydrogen isocyanide (C2H4/HNC), acetylene/hydrogen cyanide (C2H2/HCN), acetylene/hydrogen isocyanide (C2H2/HNC), and acetylene:methylenimine (C2H2:HNCH2) complexes. Ethyl isocyanide (CH3CH2NC) and a ketenimine form (CH3CH[double bond, length as m-dash]C[double bond, length as m-dash]NH) have been identified as well. Photoproduct identification and spectral assignments were done using previous studies and density functional theory (DFT) calculations with the B3LYP/cc-pVTZ basis set.

A Non-surgical Intervention for Triangular Fibrocartilage Complex Tears.

PubMed

Barlow, Susan J

2016-12-01

The current literature contains no reports of treatment options other than surgery following failed conservative management of a triangular fibrocartilage complex (TFCC) tear. The purpose of this study is to describe the use of a novel brace as a non-surgical intervention for TFCC tears. This paper is a case study of a subject with a magnetic resonance imaging-confirmed TFCC tear. As an alternative to surgery, he consented to wear a novel brace for 12 weeks after conservative management of his injury had failed. His recovery from injury was monitored with a weight-bearing tolerance test and the disabilities of the arm, shoulder and hand (DASH) outcome measure. An increase in weight-bearing tolerance and upper extremity use was evident immediately after donning the brace. After 12 weeks, the subject demonstrated a return to normal weight-bearing tolerance and normal DASH outcome measure scores. These improvements were still evident at a 1-year follow-up appointment. Utilizing this novel brace resulted in functional status improvement in a subject with a TFCC tear as demonstrated by significant changes in his DASH outcome measure scores. This case study demonstrates the first non-surgical alternative treatment for a TFCC tear after conservative management has failed. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Functional and oncological outcome after surgical resection of the scapula and clavicle for primary chondrosarcoma.

PubMed

Nota, S P F T; Russchen, M J A M; Raskin, K A; Mankin, H J; Hornicek, F J; Schwab, J H

2017-04-01

The scapula is a relatively common site for chondrosarcoma to develop in contrary to the clavicle, which is rarely affected by these tumors. The aim of this study is to determine the functional and oncological outcome for patients treated operatively for scapular or clavicular chondrosarcoma. In this single-center retrospective study, we included a sample of 20 patients that received the diagnosis of a primary chondrosarcoma of the scapula or clavicle. Of the surviving patients, the functional function was assessed using the DASH and the PROMIS Physical Function-Upper Extremity. Patients were longitudinally tracked for their oncological outcome. All patients were followed for at least 2 years or until death. The mean age of the cohort was 47 years. Eighteen patients suffered from a chondrosarcoma of the scapula, and in 2 patients, the tumor was located in the clavicle. Metastasis, local recurrence and a higher tumor grade were all associated with a decreased overall survival. For the patients with a chondrosarcoma of the scapula, the average DASH score was 16 ± 16 and the mean PROMIS Physical Function-Upper Extremity score was 48 ± 10. Patients with both an intact rotator cuff and glenoid had a better physical function. Upper extremity function after (partial) scapulectomy varied depending on whether the glenoid was spared and whether a functioning shoulder abductor remained. When the resection spared these structures, then excellent functional outcomes were reported. Oncologic outcomes depended upon the grade of the tumor and whether local recurrence and metastases occurred.

Beaten Into Submissiveness? An Investigation Into the Protective Strategies Used by Survivors of Domestic Abuse.

PubMed

Irving, Laura; Liu, Ben Chi-Pun

2016-12-01

The aim of the study was to identify the prevalence and perceived helpfulness of a variety of protective strategies that were used by female survivors of domestic abuse and to explore factors that may have influenced strategy usage. Forty participants were recruited from a voluntary sector domestic abuse service, commissioned by an outer London local authority in the United Kingdom, in early 2014. The measurement tools used were the Intimate Partner Violence Strategies Index (IPVSI) and the Coordinated Action Against Domestic Abuse (CAADA) Domestic Abuse, Stalking and "Honour"-Based Violence (DASH) Risk Assessment Checklist. The average age was 33 years ( SD = 7.9, range = 20-57); half reported to be of Asian ethnicity, 37.5% White, and 12.5% Black or Mixed ethnicity. The average DASH score was 9.8 ( SD = 13.2, range = 0-18), and an average of 18 ( SD = 6.7, range = 1-29) protective strategies were utilized by each participant. All of the most commonly used strategies were from the placating category. Although safety planning strategies were rated as the most helpful by all participants, placating strategies were also rated as helpful by two thirds of participants. Stepwise multiple regression showed that placating was the only significant predictor of DASH score (β = .375, p < .05) and accounted for 14% of the variance of DASH score. Findings showed that women utilized a diverse range of protective strategies with placating strategies being most intensely used and rated as helpful. However, placating strategy usage could be a risk factor as opposed to a protective factor. This study has also demonstrated that greater placating strategies were used by White than South Asian women, and women who were employed used more formal strategies. This research has extended the knowledge base of protective strategies that professionals can draw from to underpin decisions and interventions when working with domestic abuse survivors.

Cross-cultural adaptation and validation of the Korean version of the Oxford shoulder score.

PubMed

Roh, Young Hak; Noh, Jung Ho; Kim, Woo; Oh, Joo Han; Gong, Hyun Sik; Baek, Goo Hyun

2012-01-01

The Oxford shoulder score (OSS) is being used increasingly and has been adapted cross-culturally in some Western countries. On the other hand, there are few validated translations of the OSS in Asian countries. This study translated and adapted cross-culturally the original OSS to produce a Korean version, and assessed the validity and reliability of the Korean version of the OSS (Korean OSS). One hundred and five patients with shoulder pain caused by degenerative or inflammatory disorders completed the Korean OSS and Korean disability of arm, shoulder and hand (DASH). In addition, the pain score by a visual analog scale (VAS) during activity and at rest, subjective assessment of activities of daily living (ADL), the active range of motion (ROM), and measurements of the abduction strength (strength) were included in the validation process. There were no major linguistic or cultural problems during the forward and backward translations of the MHQ, except for a minor change due to cultural discrepancies in eating such as using a spoon and chopsticks by one dominant hand instead of a knife and fork by two hands. The internal consistency was high (Cronbach's alpha 0.91). The reproducibility test showed no significant difference (Intra-class coefficient 0.95). The construct validity, which was tested by the Pearson correlation coefficient revealed a strong correlation (r > 0.6) between the Korean OSS against subscale of DASH disability/symptom, DASH work and ADL, as well as a moderate correlation (0.3 < r < 0.6) with the DASH sports/music, strength, ROM, pain during activity and pain at rest. The Korean OSS proved to be valid by demonstrating a significant correlation with the patient-based upper extremity questionnaire and clinical assessment. The application and evaluation of the instrument is feasible and understandable among patients in Korea.

A method of coupling the Paternò-Büchi reaction with direct infusion ESI-MS/MS for locating the C[double bond, length as m-dash]C bond in glycerophospholipids.

PubMed

Stinson, Craig A; Xia, Yu

2016-06-21

Tandem mass spectrometry (MS/MS) coupled with soft ionization is established as an essential platform for lipid analysis; however, determining high order structural information, such as the carbon-carbon double bond (C[double bond, length as m-dash]C) location, remains challenging. Recently, our group demonstrated a method for sensitive and confident lipid C[double bond, length as m-dash]C location determination by coupling online the Paternò-Büchi (PB) reaction with nanoelectrospray ionization (nanoESI) and MS/MS. Herein, we aimed to expand the scope of the PB reaction for lipid analysis by enabling the reaction with infusion ESI-MS/MS at much higher flow rates than demonstrated in the nanoESI setup (∼20 nL min(-1)). In the new design, the PB reaction was effected in a fused silica capillary solution transfer line, which also served as a microflow UV reactor, prior to ESI. This setup allowed PB reaction optimization and kinetics studies. Under optimized conditions, a maximum of 50% PB reaction yield could be achieved for a standard glycerophosphocholine (PC) within 6 s of UV exposure over a wide flow rate range (0.1-10 μL min(-1)). A solvent composition of 7 : 3 acetone : H2O (with 1% acid or base modifier) allowed the highest PB yields and good lipid ionization, while lower yields were obtained with an addition of a variety of organic solvents. Radical induced lipid peroxidation was identified to induce undesirable side reactions, which could be effectively suppressed by eliminating trace oxygen in the solution via N2 purge. Finally, the utility of coupling the PB reaction with infusion ESI-MS/MS was demonstrated by analyzing a yeast polar lipid extract where C[double bond, length as m-dash]C bond locations were revealed for 35 glycerophospholipids (GPs).

Positive effect of an autologous platelet concentrate in lateral epicondylitis in a double-blind randomized controlled trial: platelet-rich plasma versus corticosteroid injection with a 1-year follow-up.

PubMed

Peerbooms, Joost C; Sluimer, Jordi; Bruijn, Daniël J; Gosens, Taco

2010-02-01

Platelet-rich plasma (PRP) has shown to be a general stimulation for repair. Purpose To determine the effectiveness of PRP compared with corticosteroid injections in patients with chronic lateral epicondylitis. Randomized controlled trial; Level of evidence, 1. The trial was conducted in 2 teaching hospitals in the Netherlands. One hundred patients with chronic lateral epicondylitis were randomly assigned in the PRP group (n = 51) or the corticosteroid group (n = 49). A central computer system carried out randomization and allocation to the trial group. Patients were randomized to receive either a corticosteroid injection or an autologous platelet concentrate injection through a peppering technique. The primary analysis included visual analog scores and DASH Outcome Measure scores (DASH: Disabilities of the Arm, Shoulder, and Hand). Successful treatment was defined as more than a 25% reduction in visual analog score or DASH score without a reintervention after 1 year. The results showed that, according to the visual analog scores, 24 of the 49 patients (49%) in the corticosteroid group and 37 of the 51 patients (73%) in the PRP group were successful, which was significantly different (P <.001). Furthermore, according to the DASH scores, 25 of the 49 patients (51%) in the corticosteroid group and 37 of the 51 patients (73%) in the PRP group were successful, which was also significantly different (P = .005). The corticosteroid group was better initially and then declined, whereas the PRP group progressively improved. Treatment of patients with chronic lateral epicondylitis with PRP reduces pain and significantly increases function, exceeding the effect of corticosteroid injection. Future decisions for application of the PRP for lateral epicondylitis should be confirmed by further follow-up from this trial and should take into account possible costs and harms as well as benefits.

Platelet-rich plasma versus autologous blood versus steroid injection in lateral epicondylitis: systematic review and network meta-analysis.

PubMed

Arirachakaran, Alisara; Sukthuayat, Amnat; Sisayanarane, Thaworn; Laoratanavoraphong, Sorawut; Kanchanatawan, Wichan; Kongtharvonskul, Jatupon

2016-06-01

Clinical outcomes between the use of platelet-rich plasma (PRP), autologous blood (AB) and corticosteroid (CS) injection in lateral epicondylitis are still controversial. A systematic review and network meta-analysis of randomized controlled trials was conducted with the aim of comparing relevant clinical outcomes between the use of PRP, AB and CS injection. Medline and Scopus databases were searched from inception to January 2015. A network meta-analysis was performed by applying weight regression for continuous outcomes and a mixed-effect Poisson regression for dichotomous outcomes. Ten of 374 identified studies were eligible. When compared to CS, AB injection showed significantly improved effects with unstandardized mean differences (UMD) in pain visual analog scale (VAS), Disabilities of Arm Shoulder and Hand (DASH), Patient-Related Tennis Elbow Evaluation (PRTEE) score and pressure pain threshold (PPT) of -2.5 (95 % confidence interval, -3.5, -1.5), -25.5 (-33.8, -17.2), -5.3 (-9.1, -1.6) and 9.9 (5.6, 14.2), respectively. PRP injections also showed significantly improved VAS and DASH scores when compared with CS. PRP showed significantly better VAS with UMD when compared to AB injection. AB injection has a higher risk of adverse effects, with a relative risk of 1.78 (1.00, 3.17), when compared to CS. The network meta-analysis suggested no statistically significant difference in multiple active treatment comparisons of VAS, DASH and PRTEE when comparing PRP and AB injections. However, AB injection had improved DASH score and PPT when compared with PRP injection. In terms of adverse effects, AB injection had a higher risk than PRP injection. This network meta-analysis provided additional information that PRP injection can improve pain and lower the risk of complications, whereas AB injection can improve pain, disabilities scores and pressure pain threshold but has a higher risk of complications. Level I evidence.

Ongoing positive effect of platelet-rich plasma versus corticosteroid injection in lateral epicondylitis: a double-blind randomized controlled trial with 2-year follow-up.

PubMed

Gosens, Taco; Peerbooms, Joost C; van Laar, Wilbert; den Oudsten, Brenda L

2011-06-01

Platelet-rich plasma (PRP) has been shown to be a general stimulation for repair and 1-year results showed promising success percentages. This trial was undertaken to determine the effectiveness of PRP compared with corticosteroid injections in patients with chronic lateral epicondylitis with a 2-year follow-up. Randomized controlled trial; Level of evidence, 1. The trial was conducted in 2 Dutch teaching hospitals. One hundred patients with chronic lateral epicondylitis were randomly assigned to a leukocyte-enriched PRP group (n = 51) or the corticosteroid group (n = 49). Randomization and allocation to the trial group were carried out by a central computer system. Patients received either a corticosteroid injection or an autologous platelet concentrate injection through a peppering needling technique. The primary analysis included visual analog scale (VAS) pain scores and Disabilities of the Arm, Shoulder and Hand (DASH) outcome scores. The PRP group was more often successfully treated than the corticosteroid group (P < .0001). Success was defined as a reduction of 25% on VAS or DASH scores without a reintervention after 2 years. When baseline VAS and DASH scores were compared with the scores at 2-year follow-up, both groups significantly improved across time (intention-to-treat principle). However, the DASH scores of the corticosteroid group returned to baseline levels, while those of the PRP group significantly improved (as-treated principle). There were no complications related to the use of PRP. Treatment of patients with chronic lateral epicondylitis with PRP reduces pain and increases function significantly, exceeding the effect of corticosteroid injection even after a follow-up of 2 years. Future decisions for application of PRP for lateral epicondylitis should be confirmed by further follow-up from this trial and should take into account possible costs and harms as well as benefits.

The role of Dietary Approaches to Stop Hypertension (DASH) diet food groups in blood pressure in type 2 diabetes.

PubMed

de Paula, Tatiana Pedroso; Steemburgo, Thais; de Almeida, Jussara Carnevale; Dall'Alba, Valesca; Gross, Jorge Luiz; de Azevedo, Mirela Jobim

2012-07-14

The role of each Dietary Approaches to Stop Hypertension (DASH) diet component in blood pressure (BP) of patients with diabetes is still uncertain. The aim of the present study was to evaluate possible associations of the recommended food groups of the DASH diet eating plan with BP values in patients with type 2 diabetes. In the present cross-sectional study, 225 patients with type 2 diabetes (age 61·1 (SD 10·4) years; diabetes duration 13·1 (SD 9·1) years; males 48·4 %; BMI 28·5 (SD 4·3) kg/m(2); HbA1c 7·1 (SD 1·3) %; systolic BP 136·7 (SD 20·0) mmHg; diastolic BP 78·4 (SD 11·8) mmHg) without dietary counselling during the previous 6 months had their dietary intake assessed by 3 d weighed-diet records. Patients were divided into two groups according to BP tertiles: LOW BP (first tertile) and HIGH BP (second plus third tertiles). Multivariate logistic regression models demonstrated that the daily intake of 80 g of fruits per 4184 kJ (1000 kcal) (OR 0·781; 95 % CI 0·617, 0·987; P = 0·039) or 50 g of vegetables per 4184 kJ (1000 kcal) (OR 0·781; 95 % CI 0·618, 0·988; P = 0·040) reduced the chance of the presence of HIGH mean BP (MBP ≥ 92 mmHg) by 22 % each, adjusted for possible confounders. In conclusion, fruit and vegetables were the food groups of the DASH diet associated with reduced BP values in patients with type 2 diabetes, and their consumption might play a protective role against increased BP values.

Intra-urban spatial variability of PM2.5-bound carbonaceous components

NASA Astrophysics Data System (ADS)

Xie, Mingjie; Coons, Teresa L.; Dutton, Steven J.; Milford, Jana B.; Miller, Shelly L.; Peel, Jennifer L.; Vedal, Sverre; Hannigan, Michael P.

2012-12-01

The Denver Aerosol Sources and Health (DASH) study was designed to evaluate associations between PM2.5 species and sources and adverse human health effects. The DASH study generated a five-year (2003-2007) time series of daily speciated PM2.5 concentration measurements from a single, special-purpose monitoring site in Denver, CO. To evaluate the ability of this site to adequately represent the short term temporal variability of PM2.5 concentrations in the five county Denver metropolitan area, a one year supplemental set of PM2.5 samples was collected every sixth day at the original DASH monitoring site and concurrently at three additional sites. Two of the four sites, including the original DASH site, were located in residential areas at least 1.9 km from interstate highways. The other two sites were located within 0.3 km of interstate highways. Concentrations of elemental carbon (EC), organic carbon (OC), and 58 organic molecular markers were measured at each site. To assess spatial variability, site pairs were compared using the Pearson correlation coefficient (r) and coefficient of divergence (COD), a statistic that provides information on the degree of uniformity between monitoring sites. Bi-weekly co-located samples collected from July 2004 to September 2005 were also analyzed and used to estimate the uncertainty associated with sampling and analytical measurement for each species. In general, the two near-highway sites exhibited higher concentrations of EC, OC, polycyclic aromatic hydrocarbons (PAHs), and steranes than did the more residential sites. Lower spatial heterogeneity based on r and COD was inferred for all carbonaceous species after considering their divergence and lack of perfect correlations in co-located samples. Ratio-ratio plots combined with available gasoline- and diesel-powered motor vehicle emissions profiles for the region suggested a greater impact to high molecular weight (HMW) PAHs from diesel-powered vehicles at the near-highway sites and a more uniformly distributed impact to ambient hopanes from gasoline-powered motor vehicles at all four sites.

Utilization of Away-From-Home Food Establishments, Dietary Approaches to Stop Hypertension Dietary Pattern, and Obesity.

PubMed

Penney, Tarra L; Jones, Nicholas R V; Adams, Jean; Maguire, Eva R; Burgoine, Thomas; Monsivais, Pablo

2017-11-01

Eating meals away from home has been associated with the consumption of unhealthy foods and increased body weight. However, more rigorous assessment of the contribution of different types of away-from-home food establishments to overall diet quality and obesity is minimal. This study examined usage of these food establishments, accordance to the Dietary Approaches to Stop Hypertension (DASH) dietary pattern and obesity status in a nationally representative sample of adults in the United Kingdom. A cross-sectional analysis of data from a national survey (N=2,083 aged ≥19 years, from 2008 to 2012) with dietary intake measured using a 4-day food diary, and height and weight measured objectively. Exposures included usage of (i.e., by proportion of energy) all away-from-home food establishments combined, and fast-food outlets, restaurants, and cafés separately. Outcomes included accordance with the DASH diet, and obesity status. Multivariable logistic regressions were conducted in 2016 to estimate associations between food establishments, diet quality, and obesity. People consuming a higher proportion of energy from any away-from-home food establishment had lower odds of DASH accordance (OR=0.45, 95% CI=0.31, 0.67) and increased odds of obesity (OR=1.48, 95% CI=1.10, 1.99). After adjustment, only use of fast-food outlets was significantly associated with lower odds of DASH accordance (OR=0.48, 95% CI=0.33, 0.69) and higher odds of obesity (OR=1.30, 95% CI=1.01, 1.69). Although a greater reliance on eating away-from-home is associated with less-healthy diets and obesity, dietary public health interventions that target these food establishments may be most effective if they focus on modifying the use of fast-food outlets. Copyright © 2017 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights reserved.

Dietary Patterns Over Time and Microalbuminuria in Youth and Young Adults With Type 1 Diabetes: The SEARCH Nutrition Ancillary Study.

PubMed

Costacou, Tina; Crandell, Jamie; Kahkoska, Anna R; Liese, Angela D; Dabelea, Dana; Lawrence, Jean M; Pettitt, David J; Reynolds, Kristi; Mayer-Davis, Elizabeth J; Mottl, Amy K

2018-06-14

We assessed the association between diet quality and microalbuminuria in youth-onset type 1 diabetes using three indices: a modified Mediterranean diet score for children and adolescents (mKIDMED), the Dietary Approaches to Stop Hypertension (DASH), and the Healthy Eating Index-2010 (HEI). Youth and young adults from the SEARCH (Search for Diabetes in Youth) Nutrition Ancillary Study diagnosed with type 1 diabetes in 2002-2008, who had repeated dietary assessments at baseline and follow-up visits and urine albumin-to-creatinine ratio (UACR) measured at the outcome visit (2012-2015) ( n = 461), were selected for study. Regression models estimated the association between each longitudinally assessed diet score and UACR and microalbuminuria (UACR ≥30 μg/mg). The cohort was 43% female, and at follow-up, mean age was 20 years, disease duration was 108 months, and 7% had microalbuminuria. Adherence to a higher-quality diet was low for the mKIDMED (mean 3.7 of a possible range of -3 to 12) and the DASH (mean 42 of 80) and better, for the HEI (mean 56.3 of 100). A borderline inverse association was observed between the HEI score and microalbuminuria after adjustment for caloric and protein intake and demographic and disease factors (odds ratio [OR] HEI 0.83, P = 0.07), which lost significance with further adjustment for HbA 1c and systolic blood pressure (OR HEI 0.86, P = 0.19). Results were similar for continuous UACR. No significant associations were observed for diet quality characterized by the mKIDMED or DASH indices. Greater adherence to the HEI may be beneficial for kidney health in youth and young adults with type 1 diabetes. Low adherence to the mKIDMED and DASH diets may explain the lack of association with microalbuminuria. © 2018 by the American Diabetes Association.

Return to competitive sports after medial epicondyle fractures in adolescent athletes: results of operative and nonoperative treatment.

PubMed

Lawrence, J Todd R; Patel, Neeraj M; Macknin, Jonathan; Flynn, John M; Cameron, Danielle; Wolfgruber, Hayley C; Ganley, Theodore J

2013-05-01

The optimal treatment of medial epicondyle fractures in pediatric athletes remains unclear. To evaluate the outcomes of operative and nonoperative management of medial epicondyle fractures in young athletes. Case series; Level of evidence, 4. The records of all children with fractures of the medial epicondyle over a 5-year period, with a minimum 2 years of follow-up at a pediatric tertiary referral center, were reviewed. Patients with intra-articular entrapment of the fracture fragment or ulnar nerve entrapment were excluded. Treatment decisions were made primarily based on injury mechanism and elbow laxity or instability. Patients were contacted and asked to complete a modified Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire. Complete data with 2-year follow-up were available for 20 athletes: 6 treated nonoperatively and 14 treated operatively. At the latest follow-up, both groups achieved excellent DASH scores. Half of each cohort required physical therapy, and 6 of 14 patients who received operative treatment reported numbness. All patients were either very or completely satisfied with their treatment. Fourteen patients were overhead athletes (8 treated operatively, 6 nonoperatively). Excellent DASH scores were achieved in both groups, and all overhead athletes were able to return to their sport at the next appropriate level. Seven patients were baseball pitchers and sustained a fracture while throwing (4 treated operatively, 3 nonoperatively). None felt their performance was limited after treatment, and excellent DASH scores were achieved in both groups. These data demonstrate that nonoperative treatment can be successful in young athletes with low-energy medial epicondyle avulsions, a stable elbow, and minimal fracture displacement. Surgical management can be successful in athletes who sustain more significant trauma, who have elbow laxity or instability, or who have significant fracture fragment displacement after a fracture of the medial epicondyle.

Purification, characterization and molecular cloning of alpha-2-macroglobulin in cobia, Rachycentron canadum.

PubMed

Chuang, Wen-Hsiao; Liu, Ping-Chung; Hung, Chia-Yu; Lee, Kuo-Kau

2014-12-01

Alpha-2-macroglobulin (α-2-M) is a broad spectrum protease inhibitor which is abundant in the plasma of vertebrates and several invertebrates. The α-2-M was purified from cobia (Rachycentron canadum) plasma by a four-step procedure: poly ethylene glycol fractionation, affinity chromatography, hydrophobic interaction chromatography and ion exchange chromatography on Fast Protein liquid chromatography system in the present study. It migrated as one protein band with a molecular mass of about 360 kDa in the native state, whereas in SDS-PAGE it was about 180 kDa under non-reducing condition. This result revealed that the native protein was a dimer. In addition, it was cleaved into two different fragments of molecular mass about 93 and 87 kDa when reduced by dithiothreitol (DTT). The anti-protease activity of the purified α-2-M was apparently decreased as temperature elevated above 50 °C. The α-2-M exhibited highest protease inhibitory activity at pH 9. The results indicate that the α-2-M is a heat-labile and alkaline protease inhibitor. The purified α-2-M exhibited more than 50% protease inhibitory activity against extracellular products (ECP) of Vibrio alginolytius isolated from diseased cobia. It seems that the protease activities in ECP may be affected by the plasma α-2-M. The protease inhibitory activities of cobia plasma or purified α-2-M were decreased when incubated with 10 mM methylamine for 30 min. The α-2-M cDNA consisted of 4611 bp with an open reading frame of 4374 bp had been cloned from cobia liver. This sequence contained thioester domain (GCGEQ) and thirteen predicted N-linked glycosylation sites. In addition, the amino acid sequence of thioester domain and genes of adjacent regions of cobia α-2-M were further compared with sequences of known fish species in GenBank. The unweighted pair group method using arithmetic average (UPGMA) was employed to construct the phylogenetic trees of α-2-M among different fish species (freshwater fish, sea water fish and primitive fish), and all these fish species were then clustered into three groups. The cobia α-2-M was closer to that of sea water fish than that of freshwater fish compared basing on its similarity of amino acid sequence and phylogenetic analysis of the partial gene. Copyright © 2014 Elsevier Ltd. All rights reserved.

Synthesis, structure and reactivity of rare-earth metallacarborane alkyls [η(1):η(5)-O(CH2)2C2B9H9]Ln(σ:η(1)-CH2C6H4-o-NMe2)(THF)2.

PubMed

Yang, Jingying; Xie, Zuowei

2015-04-14

Rare-earth metallacarborane alkyls can be stabilized by the incorporation of a functional sidearm into both π and σ ligands. Reaction of [Me3NH][7,8-O(CH2)2-7,8-C2B9H10] with one equiv. of Ln(CH2C6H4-o-NMe2)3 gave metallacarborane alkyls [η(1):η(5)-O(CH2)2C2B9H9]Ln(σ:η(1)-CH2C6H4-o-NMe2)(THF)2 (Ln = Y (), Gd (), Er ()) via alkane elimination. They represent the first examples of rare-earth metallacarborane alkyls. Treatment of with RN[double bond, length as m-dash]C[double bond, length as m-dash]NR (R = Cy, (i)Pr) or 2-benzoylpyridine afforded the corresponding mono-insertion products [η(1):η(5)-O(CH2)2C2B9H9]Y[η(2)-(RN)2C(CH2C6H4-o-NMe2)](DME) (R = Cy (), (i)Pr ()) or [η(1):η(5)-O(CH2)2C2B9H9]Y[C5H4NC(Ph)(CH2C6H4-o-NMe2)O](THF)2 (), respectively. Complex also reacted with ArNCO or ArNC (Ar = 2,6-diisopropylphenyl, 2,6-dimethylphenyl) to give di-insertion products [η(1):η(5)-O(CH2)2C2B9H9]Y[OC([double bond, length as m-dash]NC6H3Me2)N(C6H3Me2)C(CH2C6H4-o-NMe2)O](THF)2 () or [η(1):η(5)-O(CH2)2C2B9H9]Y[C([double bond, length as m-dash]NC6H3(i)Pr2)C([double bond, length as m-dash]NC6H3(i)Pr2)(CH2C6H4-o-NMe2)](DME) (). These results showed that the reactivity pattern of the Ln-C σ bond in rare-earth metallacarborane alkyls was dependent on the nature of the unsaturated organic molecules. New complexes were characterized by various spectroscopic techniques and elemental analysis. Some were further confirmed by single-crystal X-ray analysis.

The Dimer Interfaces of Protease and Extra-Protease Domains Influence the Activation of Protease and the Specificity of GagPol Cleavage

PubMed Central

Pettit, Steven C.; Gulnik, Sergei; Everitt, Lori; Kaplan, Andrew H.

2003-01-01

Activation of the human immunodeficiency virus type 1 (HIV-1) protease is an essential step in viral replication. As is the case for all retroviral proteases, enzyme activation requires the formation of protease homodimers. However, little is known about the mechanisms by which retroviral proteases become active within their precursors. Using an in vitro expression system, we have examined the determinants of activation efficiency and the order of cleavage site processing for the protease of HIV-1 within the full-length GagPol precursor. Following activation, initial cleavage occurs between the viral p2 and nucleocapsid proteins. This is followed by cleavage of a novel site located in the transframe domain. Mutational analysis of the dimer interface of the protease produced differential effects on activation and specificity. A subset of mutations produced enhanced cleavage at the amino terminus of the protease, suggesting that, in the wild-type precursor, cleavages that liberate the protease are a relatively late event. Replacement of the proline residue at position 1 of the protease dimer interface resulted in altered cleavage of distal sites and suggests that this residue functions as a cis-directed specificity determinant. In summary, our studies indicate that interactions within the protease dimer interface help determine the order of precursor cleavage and contribute to the formation of extended-protease intermediates. Assembly domains within GagPol outside the protease domain also influence enzyme activation. PMID:12477841

The dimer interfaces of protease and extra-protease domains influence the activation of protease and the specificity of GagPol cleavage.

PubMed

Pettit, Steven C; Gulnik, Sergei; Everitt, Lori; Kaplan, Andrew H

2003-01-01

Activation of the human immunodeficiency virus type 1 (HIV-1) protease is an essential step in viral replication. As is the case for all retroviral proteases, enzyme activation requires the formation of protease homodimers. However, little is known about the mechanisms by which retroviral proteases become active within their precursors. Using an in vitro expression system, we have examined the determinants of activation efficiency and the order of cleavage site processing for the protease of HIV-1 within the full-length GagPol precursor. Following activation, initial cleavage occurs between the viral p2 and nucleocapsid proteins. This is followed by cleavage of a novel site located in the transframe domain. Mutational analysis of the dimer interface of the protease produced differential effects on activation and specificity. A subset of mutations produced enhanced cleavage at the amino terminus of the protease, suggesting that, in the wild-type precursor, cleavages that liberate the protease are a relatively late event. Replacement of the proline residue at position 1 of the protease dimer interface resulted in altered cleavage of distal sites and suggests that this residue functions as a cis-directed specificity determinant. In summary, our studies indicate that interactions within the protease dimer interface help determine the order of precursor cleavage and contribute to the formation of extended-protease intermediates. Assembly domains within GagPol outside the protease domain also influence enzyme activation.

The effects of adult day services on family caregivers' daily stress, affect, and health: outcomes from the Daily Stress and Health (DaSH) study.

PubMed

Zarit, Steven H; Kim, Kyungmin; Femia, Elia E; Almeida, David M; Klein, Laura C

2014-08-01

We examine the effects of use of adult day service (ADS) by caregivers of individuals with dementia (IWD) on daily stressors, affect, and health symptoms. Participants were interviewed for 8 consecutive days. On some days, the IWD attended an ADS program and on the other days caregivers provide most or all of the care at home. Participants were 173 family caregivers of IWDs using an ADS program. Daily telephone interviews assessed care-related stressors, noncare stressors, positive events, affect, and health symptoms. Multilevel models with data nested within persons were used to examine effects of ADS use on daily stressor exposure, affect, and health symptoms. Caregivers had lower exposure to care-related stressors on ADS days, more positive experiences, and more noncare stressors. ADS use lowered anger and reduced the impact of noncare stressors on depressive symptoms. The findings demonstrate that stressors on caregivers are partly lowered, and affect is improved on ADS days, which may provide protection against the effects of chronic stress associated with caregiving. © The Author 2013. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Protease production by fermentation of fish solubles from salmon canning processes.

PubMed

Wah-On, H C; Branion, R M; Strasdine, G A

1980-09-01

Production of protease by fermentation, using Sorangium 495, of a substrate based on condensed fish solubles is demonstrated. The effects of carbohydrate addition, pH, fish solubles concentration, scale-up, agitation, and air flow rate on protease yields are described. While the fish solubles medium alone could give rise to measurable yields of protease, these were, at worst, doubled when 1% glucose was added to the medium. pH 7 was optimal for protease yield. Although the concentration of fish solubles in the basic medium showed no significant effect on cell yield, maximum protease yield was observed at a protein concentration equivalent to 3.85 mg/mL of bovine serum albumin. Protease production rates decreased as medium protein fermentor showed no significant effect on maximum protease yields. The effects of agitator speed and air flow rate on protease yield suggested that the rate of O2 transfer from air to medium could limit the rate of protease production. It was also noted that protease production is not growth associated.

Dynamic characteristics of undoped and p-doped Fabry-Perot InAs/InP quantum dash based ridge waveguide lasers for access/metro networks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mollet, O., E-mail: oriane.mollet@lpn.cnrs.fr; Martinez, A.; Merghem, K.

In this paper, we report the characteristics of InAs/InP quantum dashes (QDash) based lasers emitting around 1.55 μm. An unprecedented high modal gain of ∼100 cm{sup −1} is obtained for an optimized active structure by stacking 12 QDash layers. Directly modulated lasers allowed achieving a modulation bandwidth of ∼10 GHz and a Henry factor around 5. Thanks to p-type doping, the Henry factor value is reduced down to 2.7 while the modulation bandwidth still amounts to ∼10 GHz. This shows that doping of the active region is important to improve the dynamic characteristics of QDash lasers.

The proteolytic system of pineapple stems revisited: Purification and characterization of multiple catalytically active forms.

PubMed

Matagne, André; Bolle, Laetitia; El Mahyaoui, Rachida; Baeyens-Volant, Danielle; Azarkan, Mohamed

2017-06-01

Crude pineapple proteases extract (aka stem bromelain; EC 3.4.22.4) is an important proteolytic mixture that contains enzymes belonging to the cysteine proteases of the papain family. Numerous studies have been reported aiming at the fractionation and characterization of the many molecular species present in the extract, but more efforts are still required to obtain sufficient quantities of the various purified protease forms for detailed physicochemical, enzymatic and structural characterization. In this work, we describe an efficient strategy towards the purification of at least eight enzymatic forms. Thus, following rapid fractionation on a SP-Sepharose FF column, two sub-populations with proteolytic activity were obtained: the unbound (termed acidic) and bound (termed basic) bromelain fractions. Following reversible modification with monomethoxypolyethylene glycol (mPEG), both fractions were further separated on Q-Sepharose FF and SP-Sepharose FF, respectively. This procedure yielded highly purified molecular species, all titrating ca. 1 mol of thiol group per mole of enzyme, with distinct biochemical properties. N-terminal sequencing allowed identifying at least eight forms with proteolytic activity. The basic fraction contained previously identified species, i.e. basic bromelain forms 1 and 2, ananain forms 1 and 2, and comosain (MEROPS identifier: C01.027). Furthermore, a new proteolytic species, showing similarities with basic bomelain forms 1 and 2, was discovered and termed bromelain form 3. The two remaining species were found in the acidic bromelain fraction and were arbitrarily named acidic bromelain forms 1 and 2. Both, acidic bromelain forms 1, 2 and basic bromelain forms 1, 2 and 3 are glycosylated, while ananain forms 1 and 2, and comosain are not. The eight protease forms display different amidase activities against the various substrates tested, namely small synthetic chromogenic compounds (DL-BAPNA and Boc-Ala-Ala-Gly-pNA), fluorogenic compounds (like Boc-Gln-Ala-Arg-AMC, Z-Arg-Arg-AMC and Z-Phe-Arg-AMC), and proteins (azocasein and azoalbumin), suggesting a specific organization of their catalytic residues. All forms are completely inhibited by specific cysteine and cysteine/serine protease inhibitors, but not by specific serine and aspartic protease inhibitors, with the sole exception of pepstatin A that significantly affects acidic bromelain forms 1 and 2. For all eight protease forms, inhibition is also observed with 1,10-phenanthrolin, a metalloprotease inhibitor. Metal ions (i.e. Mn 2+ , Mg 2+ and Ca 2+ ) showed various effects depending on the protease under consideration, but all of them are totally inhibited in the presence of Zn 2+ . Mass spectrometry analyses revealed that all forms have a molecular mass of ca. 24 kDa, which is characteristic of enzymes belonging to the papain-like proteases family. Far-UV CD spectra analysis further supported this analysis. Interestingly, secondary structure calculation proves to be highly reproducible for all cysteine proteases of the papain family tested so far (this work; see also Azarkan et al., 2011; Baeyens-Volant et al., 2015) and thus can be used as a test for rapid identification of the classical papain fold. Copyright © 2017 Elsevier Ltd. All rights reserved.

Protease inhibitor from Moringa oleifera with potential for use as therapeutic drug and as seafood preservative

PubMed Central

Bijina, B.; Chellappan, Sreeja; Krishna, Jissa G.; Basheer, Soorej M.; Elyas, K.K.; Bahkali, Ali H.; Chandrasekaran, M.

2011-01-01

Protease inhibitors are well known to have several applications in medicine and biotechnology. Several plant sources are known to return potential protease inhibitors. In this study plants belonging to different families of Leguminosae, Malvaceae, Rutaceae, Graminae and Moringaceae were screened for the protease inhibitor. Among them Moringa oleifera, belonging to the family Moringaceae, recorded high level of protease inhibitor activity after ammonium sulfate fractionation. M. oleifera, which grows throughout most of the tropics and having several industrial and medicinal uses, was selected as a source of protease inhibitor since so far no reports were made on isolation of the protease inhibitor. Among the different parts of M. oleifera tested, the crude extract isolated from the mature leaves and seeds showed the highest level of inhibition against trypsin. Among the various extraction media evaluated, the crude extract prepared in phosphate buffer showed maximum recovery of the protease inhibitor. The protease inhibitor recorded high inhibitory activity toward the serine proteases thrombin, elastase, chymotrypsin and the cysteine proteases cathepsin B and papain which have more importance in pharmaceutical industry. The protease inhibitor also showed complete inhibition of activities of the commercially available proteases of Bacillus licheniformis and Aspergillus oryzae. However, inhibitory activities toward subtilisin, esperase, pronase E and proteinase K were negligible. Further, it was found that the protease inhibitor could prevent proteolysis in a commercially valuable shrimp Penaeus monodon during storage indicating the scope for its application as a seafood preservative. This is the first report on isolation of a protease inhibitor from M. oleifera. PMID:23961135

Suppression of Calpain Expression by NSAIDs is Associated with Inhibition of Cell Migration in Rat Duodenum

PubMed Central

Silver, Kristopher; Littlejohn, A.; Thomas, Laurel; Bawa, Bhupinder; Lillich, James D.

2017-01-01

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used for the alleviation of pain and inflammation, but these drugs are also associated with a suite of negative side effects. Gastrointestinal (GI) toxicity is particularly concerning since it affects an estimated 70% of individuals taking NSAIDs routinely, and evidence suggests the majority of toxicity is occurring in the small intestine. Traditionally, NSAID-induced GI toxicity has been associated with indiscriminate inhibition of cyclooxygenase isoforms, but other mechanisms, including inhibition of cell migration, intestinal restitution, and wound healing, are likely to contribute to toxicity. Previous efforts demonstrated that treatment of cultured intestinal epithelial cells (IEC) with NSAIDs inhibits expression and activity of calpain proteases, but the effects of specific inhibition of calpain expression in vitro or the effects of NSAIDs on intestinal cell migration in vivo remain to be determined. Accordingly, we examined the effect of suppression of calpain protease expression with siRNA on cell migration in cultured IECs and evaluated the effects of NSAID treatment on epithelial cell migration and calpain protease expression in rat duodenum. Our results show that calpain siRNA inhibits protease expression and slows migration in cultured IECs. Additionally, NSAID treatment of rats slowed migration up the villus axis and suppressed calpain expression in duodenal epithelial cells. Our results are supportive of the hypothesis that suppression of calpain expression leading to slowing of cell migration is a potential mechanism through which NSAIDs cause GI toxicity. PMID:28342779

Proliferation of protease-enriched mast cells in sarcoptic skin lesions of raccoon dogs.

PubMed

Noviana, D; W Harjanti, D; Otsuka, Y; Horii, Y

2004-07-01

Skin sites, tongue, lung, liver, jejunum and rectum from two raccoon dogs with Sarcoptes scabiei infestation and five normal (control) raccoon dogs were examined in terms of the distribution, proteoglycan properties and protease activity of mast cells. Infestation with S. scabiei caused a significant increase in the number of dermal mast cells. While the number of mast cells (average +/- standard deviation) in specimens of skin from the dorsum, dorsal neck, dorsal hind foot and dorsal fore foot was 40.0 +/- 19.8/mm2 in control animals, it was 236.1 +/- 58.9/mm2 in the skin of mange-infested animals. Histochemical analysis revealed the glycosaminoglycan, heparin, within the mast cells of all organs examined in both control and affected animals. Enzyme-histochemical detection of serine proteases demonstrated an increase in mast-cell-specific protease activity (i.e., chymase and tryptase) in the skin of infested animals. The percentage of mast cells demonstrating chymase activity was 53.0 +/- 27.4% in control animals and 73.8 +/- 19.4% in mite-infested animals. The corresponding results for tryptase activity were 53.5 +/- 25.2% and 89.4 +/- 9.8%. Increases in mast cell chymase or tryptase activity, or both, were also observed within other organs of the infected animals, but the total number of mast cells found at such sites (with the exception of liver and ventrolateral pinna) did not differ from those of control animals. Copyright 2004 Elsevier Ltd.

Purification of a factor from the granules of a rat natural killer cell line (RNK) that reduces tumor cell growth and changes tumor morphology. Molecular identity with a granule serine protease (RNKP-1).

PubMed

Sayers, T J; Wiltrout, T A; Sowder, R; Munger, W L; Smyth, M J; Henderson, L E

1992-01-01

We have purified a protein from the granules of the rat NK leukemia cell line (RNK) that is cytostatic to a variety of tumor cells. This protein shows no species specificity because certain tumor cell lines of mouse, rat, and human origin were equally sensitive to its growth inhibitory effects. Treatment of sensitive cells resulted in a rounding of the cells followed by homotypic aggregation into large aggregates. The granule protein was distinct from cytolysin, Na-Cbz-Lys-thiobenzylester-esterase, or leukolexin. It had a molecular mass of 29 to 31 kDa, bound strongly to heparin, was inactivated by heating at 70 degrees C for 5 min or reduction, but was stable to trypsin treatment. By using molecular sieve chromatography, heparin agarose chromatography, and reverse phase HPLC, this protein was purified to homogeneity. The first 33 amino acids of the N-terminal amino acid sequence showed complete identity to the sequence predicted from a rat serine protease gene recently cloned and designated RNKP-1. Therefore we have purified a novel serine protease and demonstrated that it has effects on the growth and morphology of certain tumor cells. Other serine proteases that were structurally related and have substantial homology with RNKP-1 at the amino acid level showed neither growth inhibitory properties nor affected the morphology of the tumor target cells we used.

Proteases of Wood Rot Fungi with Emphasis on the Genus Pleurotus

PubMed Central

Inácio, Fabíola Dorneles; Ferreira, Roselene Oliveira; de Araujo, Caroline Aparecida Vaz; Peralta, Rosane Marina; de Souza, Cristina Giatti Marques

2015-01-01

Proteases are present in all living organisms and they play an important role in physiological conditions. Cell growth and death, blood clotting, and immune defense are all examples of the importance of proteases in maintaining homeostasis. There is growing interest in proteases due to their use for industrial purposes. The search for proteases with specific characteristics is designed to reduce production costs and to find suitable properties for certain industrial sectors, as well as good producing organisms. Ninety percent of commercialized proteases are obtained from microbial sources and proteases from macromycetes have recently gained prominence in the search for new enzymes with specific characteristics. The production of proteases from saprophytic basidiomycetes has led to the identification of various classes of proteases. The genus Pleurotus has been extensively studied because of its ligninolytic enzymes. The characteristics of this genus are easy cultivation techniques, high yield, low nutrient requirements, and excellent adaptation. There are few studies in the literature about proteases of Pleurotus spp. This review gathers together information about proteases, especially those derived from basidiomycetes, and aims at stimulating further research about fungal proteases because of their physiological importance and their application in various industries such as biotechnology and medicine. PMID:26180792

Functional Proteomic Profiling of Secreted Serine Proteases in Health and Inflammatory Bowel Disease.

PubMed

Denadai-Souza, Alexandre; Bonnart, Chrystelle; Tapias, Núria Solà; Marcellin, Marlène; Gilmore, Brendan; Alric, Laurent; Bonnet, Delphine; Burlet-Schiltz, Odile; Hollenberg, Morley D; Vergnolle, Nathalie; Deraison, Céline

2018-05-18

While proteases are essential in gastrointestinal physiology, accumulating evidence indicates that dysregulated proteolysis plays a pivotal role in the pathophysiology of inflammatory bowel disease (IBD). Nonetheless, the identity of overactive proteases released by human colonic mucosa remains largely unknown. Studies of protease abundance have primarily investigated expression profiles, not taking into account their enzymatic activity. Herein we have used serine protease-targeted activity-based probes (ABPs) coupled with mass spectral analysis to identify active forms of proteases secreted by the colonic mucosa of healthy controls and IBD patients. Profiling of (Pro-Lys)-ABP bound proteases revealed that most of hyperactive proteases from IBD secretome are clustered at 28-kDa. We identified seven active proteases: the serine proteases cathepsin G, plasma kallikrein, plasmin, tryptase, chymotrypsin-like elastase 3 A, and thrombin and the aminopeptidase B. Only cathepsin G and thrombin were overactive in supernatants from IBD patient tissues compared to healthy controls. Gene expression analysis highlighted the transcription of genes encoding these proteases into intestinal mucosae. The functional ABP-targeted proteomic approach that we have used to identify active proteases in human colonic samples bears directly on the understanding of the role these enzymes may play in the pathophysiology of IBD.

Proteases of Wood Rot Fungi with Emphasis on the Genus Pleurotus.

PubMed

Inácio, Fabíola Dorneles; Ferreira, Roselene Oliveira; de Araujo, Caroline Aparecida Vaz; Brugnari, Tatiane; Castoldi, Rafael; Peralta, Rosane Marina; de Souza, Cristina Giatti Marques

2015-01-01

Proteases are present in all living organisms and they play an important role in physiological conditions. Cell growth and death, blood clotting, and immune defense are all examples of the importance of proteases in maintaining homeostasis. There is growing interest in proteases due to their use for industrial purposes. The search for proteases with specific characteristics is designed to reduce production costs and to find suitable properties for certain industrial sectors, as well as good producing organisms. Ninety percent of commercialized proteases are obtained from microbial sources and proteases from macromycetes have recently gained prominence in the search for new enzymes with specific characteristics. The production of proteases from saprophytic basidiomycetes has led to the identification of various classes of proteases. The genus Pleurotus has been extensively studied because of its ligninolytic enzymes. The characteristics of this genus are easy cultivation techniques, high yield, low nutrient requirements, and excellent adaptation. There are few studies in the literature about proteases of Pleurotus spp. This review gathers together information about proteases, especially those derived from basidiomycetes, and aims at stimulating further research about fungal proteases because of their physiological importance and their application in various industries such as biotechnology and medicine.

Mosaic serine proteases in the mammalian central nervous system.

PubMed

Mitsui, Shinichi; Watanabe, Yoshihisa; Yamaguchi, Tatsuyuki; Yamaguchi, Nozomi

2008-01-01

We review the structure and function of three kinds of mosaic serine proteases expressed in the mammalian central nervous system (CNS). Mosaic serine proteases have several domains in the proenzyme fragment, which modulate proteolytic function, and a protease domain at the C-terminus. Spinesin/TMPRSS5 is a transmembrane serine protease whose presynaptic distribution on motor neurons in the spinal cord suggests that it is significant for neuronal plasticity. Cell type-specific alternative splicing gives this protease diverse functions by modulating its intracellular localization. Motopsin/PRSS12 is a mosaic protease, and loss of its function causes mental retardation. Recent reports indicate the significance of this protease for cognitive function. We mention the fibrinolytic protease, tissue plasminogen activator (tPA), which has physiological and pathological functions in the CNS.

Polycystic Kidney Disease

MedlinePlus

... a healthy eating plan can help lower blood pressure. A health care provider may recommend the DASH eating plan, which focuses on fruits, vegetables, whole grains, ... Dialysis Kidney Transplant Clinical ...

In vitro digestion of soluble cashew proteins and characterization of surviving IgE-reactive peptides

USDA-ARS?s Scientific Manuscript database

The stability of food allergens to digestion varies; and the ability of food proteins to cause an allergic reaction may be affected by the susceptibility of the allergen to digestion by proteases, including pepsin and trypsin. Recent studies have demonstrated that cashew nut allergens are often a ca...

LPS induces direct death of IFN-gamma primed murine embryonic hepatocyte, BNL CL2 cells in a TNF-alpha independent manner.

PubMed

So, H S; Jung, B H; Yeum, S S; Park, J S; Kim, M S; Lee, J H; Chung, S Y; Choi, S; Chae, H J; Kim, H R; Ko, C B; Chung, H T; Park, R

2000-11-01

Although it has been well known that the role of LPS on liver damage is mediated through TNF-alpha, the mechanism by which LPS modulates the cytotoxicity of IFN-gamma on hepatocytes has not yet been clearly demonstrated. Here, we demonstrate that IFN-gamma mediated apoptosis in murine embryonic hepatocyte BNL CL2 cells is potentiated by the addition of LPS (0.5 microg/ml). Consistently, LPS markedly increases the catalytic activity of caspase 3-like protease but not caspase 1-like protease in IFN-gamma treated cells. In addition, TNF-alpha alone does not affect cell viability but rather it potentiates the cytotoxic effect of IFN-gamma on BNL CL2 cells. However, the cell viability of IFN-gamma/LPS treated cells is affected by the addition of polymyxin B but not by TNF binding protein I (TNF-BPI). These data suggest that the lipid moiety of LPS may mediate direct cytotoxicity of BNL CL2 cells in a TNF-alpha independent manner.

New insights into Clostridium perfringens epsilon toxin activation and action on the brain during enterotoxemia.

PubMed

Freedman, John C; McClane, Bruce A; Uzal, Francisco A

2016-10-01

Epsilon toxin (ETX), produced by Clostridium perfringens types B and D, is responsible for diseases that occur mostly in ruminants. ETX is produced in the form of an inactive prototoxin that becomes proteolytically-activated by several proteases. A recent ex vivo study using caprine intestinal contents demonstrated that ETX prototoxin is processed in a step-wise fashion into a stable, active ∼27 kDa band on SDS-PAGE. When characterized further by mass spectrometry, the stable ∼27 kDa band was shown to contain three ETX species with varying C-terminal residues; each of these ETX species is cytotoxic. This study also demonstrated that, in addition to trypsin and chymotrypsin, proteases such as carboxypeptidases are involved in processing ETX prototoxin. Once absorbed, activated ETX species travel to several internal organs, including the brain, where this toxin acts on the vasculature to cross the blood-brain barrier, produces perivascular edema and affects several types of brain cells including neurons, astrocytes, and oligodendrocytes. In addition to perivascular edema, affected animals show edema within the vascular walls. This edema separates the astrocytic end-feet from affected blood vessels, causing hypoxia of nervous system tissue. Astrocytes of rats and sheep affected by ETX show overexpression of aquaporin-4, a membrane channel protein that is believed to help remove water from affected perivascular spaces in an attempt to resolve the perivascular edema. Amyloid precursor protein, an early astrocyte damage indicator, is also observed in the brains of affected sheep. These results show that ETX activation in vivo seems to be more complex than previously thought and this toxin acts on the brain, affecting vascular permeability, but also damaging neurons and other cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inhibition of the mitochondrial protease, ClpP, as a therapeutic strategy for human acute myeloid leuekmia

PubMed Central

Cole, Alicia; Wang, Zezhou; Coyaud, Etienne; Voisin, Veronique; Gronda, Marcela; Jitkova, Yulia; Mattson, Rachel; Hurren, Rose; Babovic, Sonja; Maclean, Neil; Restall, Ian; Wang, Xiaoming; Jeyaraju, Danny V.; Sukhai, Mahadeo A.; Prabha, Swayam; Bashir, Shaheena; Ramakrishnan, Ashwin; Leung, Elisa; Qia, Yi Hua; Zhang, Nianxian; Combes, Kevin R.; Ketela, Troy; Lin, Fengshu; Houry, Walid A.; Aman, Ahmed; Al-awar, Rima; Zheng, Wei; Wienholds, Erno; Xu, Chang Jiang; Dick, John; Wang, Jean C.Y.; Moffat, Jason; Minden, Mark D.; Eaves, Connie J.; Bader, Gary D.; Hao, Zhenyue; Kornblau, Steven M.; Raught, Brian; Schimmer, Aaron D.

2015-01-01

Summary From an shRNA screen, we identified ClpP as a member of the mitochondrial proteome whose knockdown reduced the viability of K562 leukemic cells. Expression of this mitochondrial protease that has structural similarity to the cytoplasmic proteosome is increased in the leukemic cells from approximately half of patients with AML. Genetic or chemical inhibition of ClpP killed cells from both human AML cell lines and primary samples in which the cells showed elevated ClpP expression, but did not affect their normal counterparts. Importantly, Clpp knockout mice were viable with normal hematopoiesis. Mechanistically, we found ClpP interacts with mitochondrial respiratory chain proteins and metabolic enzymes, and knockdown of ClpP in leukemic cells inhibited oxidative phosphorylation and mitochondrial metabolism. PMID:26058080

Open reduction and internal fixation of Ideberg IV and V glenoid intra-articular fractures through a Judet approach: a retrospective analysis of 11 cases.

PubMed

Hu, Chuanzhen; Zhang, Weibin; Qin, Hui; Shen, Yuhui; Xue, Zichao; Ding, Haoliang; An, Zhiquan

2015-02-01

To evaluate the methods and the outcomes of complex intra-articular glenoid fractures, treated by open reduction and internal fixations. The outcomes of 11 cases of complex intra-articular glenoid scapular fractures were retrospectively analyzed. The fractures were classified as type IV in five cases, type Va in two and Vb in four cases, according to Ideberg classification system. The mean step or gap between the main articular fragments was 6.3 ± 6.2 (4-25) mm. The fractures were openly reduced through a Judet approach and fixed with reconstructive plates or bands placed on the lateral and medial side of affected scapula, respectively. The main articular fragments were strengthened with a 4.0-mm cannulated screw in five cases. The bone union, the anterior flexion, the external and internal rotation of the shoulders were checked and recorded. The functional outcomes were evaluated using DASH questionnaire, Constant and UCLA shoulder score systems, respectively. 11 patients were followed up with an average of 28.2 ± 12.6 (12-50) months. All the fractures were united smoothly without second intervention. At the latest visiting, the mean anterior flexion of affected shoulder was 157.3 ± 7.37° (range 150°-170°), the mean external rotation of the affected shoulder was 58.2 ± 7.5° (range 50°-70°). When the shoulder in the internal rotation, the extended thumb reached to L4 or L1 or T10 or T7 in one case, to T12 in two cases and to T8 in four cases, respectively, the mean Constant score was 91.7 ± 2.8 (86-96) points. The mean UCLA score was 32.7 ± 1.7 (30-35) points, leading to four cases of excellent and seven cases of good results. The mean DASH score was 7.4 ± 3.3 (3.4-13) points. Good outcomes could be obtained when Ideberg IV and V glenoid fractures were treated by open reduction and internal fixation through a Judet approach.

Protease and Protease-Activated Receptor-2 Signaling in the Pathogenesis of Atopic Dermatitis

PubMed Central

Lee, Sang Eun; Jeong, Se Kyoo

2010-01-01

Proteases in the skin are essential to epidermal permeability barrier homeostasis. In addition to their direct proteolytic effects, certain proteases signal to cells by activating protease-activated receptors (PARs), the G-protein-coupled receptors. The expression of functional PAR-2 on human skin and its role in inflammation, pruritus, and skin barrier homeostasis have been demonstrated. Atopic dermatitis (AD) is a multifactorial inflammatory skin disease characterized by genetic barrier defects and allergic inflammation, which is sustained by gene-environmental interactions. Recent studies have revealed aberrant expression and activation of serine proteases and PAR-2 in the lesional skin of AD patients. The imbalance between proteases and protease inhibitors associated with genetic defects in the protease/protease inhibitor encoding genes, increase in skin surface pH, and exposure to proteolytically active allergens contribute to this aberrant protease/PAR-2 signaling in AD. The increased protease activity in AD leads to abnormal desquamation, degradation of lipid-processing enzymes and antimicrobial peptides, and activation of primary cytokines, thereby leading to permeability barrier dysfunction, inflammation, and defects in the antimicrobial barrier. Moreover, up-regulated proteases stimulate PAR-2 in lesional skin of AD and lead to the production of cytokines and chemokines involved in inflammation and immune responses, itching sensation, and sustained epidermal barrier perturbation with easier allergen penetration. In addition, PAR-2 is an important sensor for exogenous danger molecules, such as exogenous proteases from various allergens, and plays an important role in AD pathogenesis. Together, these findings suggest that protease activity or PAR-2 may be a future target for therapeutic intervention for the treatment of AD. PMID:20879045

Intramembrane proteolysis: theme and variations.

PubMed

Wolfe, Michael S; Kopan, Raphael

2004-08-20

Proteases that reside in cellular membranes apparently wield water to hydrolyze the peptide bonds of substrates despite their water-excluding environment. Although these intramembrane proteases bear little or no sequence resemblance to classical water-soluble proteases, they have ostensibly converged on similar hydrolytic mechanisms. Identification of essential amino acid residues of these proteases suggests that they use residue combinations for catalysis in the same way as their soluble cousins. In contrast to classical proteases, however, the catalytic residues of intramembrane proteases lie within predicted hydrophobic transmembrane domains. Elucidating the biological functions of intramembrane proteases, identifying their substrates, and understanding how they hydrolyze peptide bonds within membranes will shed light on the ways these proteases regulate crucial biological processes and contribute to disease.

Emerging principles in protease-based drug discovery

PubMed Central

Drag, Marcin; Salvesen, Guy S.

2010-01-01

Proteases have an important role in many signalling pathways, and represent potential drug targets for diseases ranging from cardiovascular disorders to cancer, as well as for combating many parasites and viruses. Although inhibitors of well-established protease targets such as angiotensin-converting enzyme and HIV protease have shown substantial therapeutic success, developing drugs for new protease targets has proved challenging in recent years. This in part could be due to issues such as the difficulty of achieving selectivity when targeting protease active sites. This Perspective discusses the general principles in protease-based drug discovery, highlighting the lessons learned and the emerging strategies, such as targeting allosteric sites, which could help harness the therapeutic potential of new protease targets. PMID:20811381

Detergent alkaline proteases: enzymatic properties, genes, and crystal structures.

PubMed

Saeki, Katsuhisa; Ozaki, Katsuya; Kobayashi, Tohru; Ito, Susumu

2007-06-01

Subtilisin-like serine proteases from bacilli have been used in various industrial fields worldwide, particularly in the production of laundry and automatic dishwashing detergents. They belong to family A of the subtilase superfamily, which is composed of three clans, namely, true subtilisins, high-alkaline proteases, and intracellular proteases. We succeeded in the large-scale production of a high-alkaline protease (M-protease) from alkaliphilic Bacillus clausii KSM-K16, and the enzyme has been introduced into compact heavy-duty laundry detergents. We have also succeeded in the industrial-scale production of a new alkaline protease, KP-43, which was originally resistant to chemical oxidants and to surfactants, produced by alkaliphilic Bacillus sp. strain KSM-KP43 and have incorporated it into laundry detergents. KP-43 and related proteases form a new clan, oxidatively stable proteases, in subtilase family A. In this review, we describe the enzymatic properties, gene sequences, and crystal structures of M-protease, KP-43, and related enzymes.

Cloning, expression and activity analysis of a novel fibrinolytic serine protease from Arenicola cristata

NASA Astrophysics Data System (ADS)

Zhao, Chunling; Ju, Jiyu

2015-06-01

The full-length cDNA of a protease gene from a marine annelid Arenicola cristata was amplified through rapid amplification of cDNA ends technique and sequenced. The size of the cDNA was 936 bp in length, including an open reading frame encoding a polypeptide of 270 amino acid residues. The deduced amino acid sequnce consisted of pro- and mature sequences. The protease belonged to the serine protease family because it contained the highly conserved sequence GDSGGP. This protease was novel as it showed a low amino acid sequence similarity (< 40%) to other serine proteases. The gene encoding the active form of A. cristata serine protease was cloned and expressed in E. coli. Purified recombinant protease in a supernatant could dissolve an artificial fibrin plate with plasminogen-rich fibrin, whereas the plasminogen-free fibrin showed no clear zone caused by hydrolysis. This result suggested that the recombinant protease showed an indirect fibrinolytic activity of dissolving fibrin, and was probably a plasminogen activator. A rat model with venous thrombosis was established to demonstrate that the recombinant protease could also hydrolyze blood clot in vivo. Therefore, this recombinant protease may be used as a thrombolytic agent for thrombosis treatment. To our knowledge, this study is the first of reporting the fibrinolytic serine protease gene in A. cristata.

Mitochondrial enzymes are protected from stress-induced aggregation by mitochondrial chaperones and the Pim1/LON protease

PubMed Central

Bender, Tom; Lewrenz, Ilka; Franken, Sebastian; Baitzel, Catherina; Voos, Wolfgang

2011-01-01

Proteins in a natural environment are constantly challenged by stress conditions, causing their destabilization, unfolding, and, ultimately, aggregation. Protein aggregation has been associated with a wide variety of pathological conditions, especially neurodegenerative disorders, stressing the importance of adequate cellular protein quality control measures to counteract aggregate formation. To secure protein homeostasis, mitochondria contain an elaborate protein quality control system, consisting of chaperones and ATP-dependent proteases. To determine the effects of protein aggregation on the functional integrity of mitochondria, we set out to identify aggregation-prone endogenous mitochondrial proteins. We could show that major metabolic pathways in mitochondria were affected by the aggregation of key enzyme components, which were largely inactivated after heat stress. Furthermore, treatment with elevated levels of reactive oxygen species strongly influenced the aggregation behavior, in particular in combination with elevated temperatures. Using specific chaperone mutant strains, we showed a protective effect of the mitochondrial Hsp70 and Hsp60 chaperone systems. Moreover, accumulation of aggregated polypeptides was strongly decreased by the AAA-protease Pim1/LON. We therefore propose that the proteolytic breakdown of aggregation-prone polypeptides represents a major protective strategy to prevent the in vivo formation of aggregates in mitochondria. PMID:21209324

Pressure-induced structural transition of mature HIV-1 protease from a combined NMR/MD simulation approach.

PubMed

Roche, Julien; Louis, John M; Bax, Ad; Best, Robert B

2015-12-01

We investigate the pressure-induced structural changes in the mature human immunodeficiency virus type 1 protease dimer, using residual dipolar coupling (RDC) measurements in a weakly oriented solution. (1)DNH RDCs were measured under high-pressure conditions for an inhibitor-free PR and an inhibitor-bound complex, as well as for an inhibitor-free multidrug resistant protease bearing 20 mutations (PR20). While PR20 and the inhibitor-bound PR were little affected by pressure, inhibitor-free PR showed significant differences in the RDCs measured at 600 bar compared with 1 bar. The structural basis of such changes was investigated by MD simulations using the experimental RDC restraints, revealing substantial conformational perturbations, specifically a partial opening of the flaps and the penetration of water molecules into the hydrophobic core of the subunits at high pressure. This study highlights the exquisite sensitivity of RDCs to pressure-induced conformational changes and illustrates how RDCs combined with MD simulations can be used to determine the structural properties of metastable intermediate states on the folding energy landscape. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

Cross-cultural adaptation of the Korean version of the Boston carpal tunnel questionnaire: its clinical evaluation in patients with carpal tunnel syndrome following local corticosteroid injection.

PubMed

Park, Dong-Jin; Kang, Ji-Hyoun; Lee, Jeong-Won; Lee, Kyung-Eun; Wen, Lihui; Kim, Tae-Jong; Park, Yong-Wook; Nam, Tai-Seung; Kim, Myung-Sun; Lee, Shin-Seok

2013-07-01

The aim of this study was to assess and validate the Korean version of the Boston Carpal Tunnel Questionnaire (K-BCTQ) in patients with carpal tunnel syndrome (CTS). After translation and cultural adaptation of the BCTQ to a Korean version, the K-BCTQ was administered to 54 patients with CTS; it was administered again after 2 weeks to assess reliability. Additionally, we administered K-DASH and EQ-5D to assess construct-validity. In a prospective study of responsiveness to clinical change, 29 of 54 patients were treated by ultrasonography-guided local corticosteroid injection therapy. The internal consistency of the K-BCTQ was high (Cronbach's alpha: 0.915) and the intra-class correlation coefficients were 0.931 for the symptom severity scale (P<0.001) and 0.844 for the functional severity scale (P<0.001). The construct-validity between the symptom severity scale and the K-DASH, and between the functional severity scale and the K-DASH were significantly correlated (both P<0.001). Clinical improvement was noted in 29 patients with injection therapy. The effect size of symptom severity was 0.67, and that of functional severity was 0.58. In conclusion, the K-BCTQ shows good reliability, construct-validity, and acceptable responsiveness after local corticosteroid injection therapy (Clinical trial number, KCT0000050).

Stroke and Nutrition: A Review of Studies

PubMed Central

Foroughi, Mehdi; Akhavanzanjani, Mohsen; Maghsoudi, Zahra; Ghiasvand, Reza; Khorvash, Fariborz; Askari, Gholamreza

2013-01-01

Background: Stroke is one of the leading causes of death and certainly the major cause of disability in the world. Diet and nutrient has an effective role in prevention and control of the risk of stroke. The aim of this study was to review the studies on the relationship between dietary intake and stroke incidence. Methods: In this study, the terms of “Fat”, “cholesterol”, “antioxidant”, “vitamins”, “salt”, “potassium”, “calcium”, “carbohydrate”, “vegetables”, “fruits”, “meat”, “tea”, “whole grains”, “sugar-sweetened beverages”, “Mediterranean diet”, “dietary approaches to stop hypertension diet (DASH diet)”, “Western diet”, and “stroke” were searched in Pubmed search engine. The observational studies, cohort studies, clinical trial studies, systemic review, and meta-analysis reviews are also included in this study. Results: The study revealed that adherence to theimprovements in nutrition and diet canreducethe incidence ofstroke. Higher antioxidant, vitamins, potassium, calcium, vegetables, fruits, whole grain intake, and adherence to the Mediterranean dietor DASH diet can lower stroke incidence. Conclusions: Adherence to Mediterranean diet or DASH diet and increasing the consumption of antioxidant, vitamins, potassium, calcium food sources, vegetables, fruits, and whole grains intake can lower the risk of stroke. Healthy diet is effective in reducing risk of stroke, however, more studies need to be carried out in this area. PMID:23776719

Deep Friction Massage Versus Steroid Injection in the Treatment of Lateral Epicondylitis.

PubMed

Yi, Rosemary; Bratchenko, Walter W; Tan, Virak

2018-01-01

The aim of the study was to determine the efficacy of deep friction massage in the treatment of lateral epicondylitis by comparing outcomes with a control group treated with splinting and therapy and with an experimental group receiving a local steroid injection. A randomized clinical trial was conducted to compare outcomes after recruitment of consecutive patients presenting with lateral epicondylitis. Patients were randomized to receive one of 3 treatments: group 1: splinting and stretching, group 2: a cortisone injection, or group 3: a lidocaine injection with deep friction massage. Pretreatment and posttreatment parameters of visual analog scale (VAS) pain ratings, Disabilities of the Arm, Shoulder and Hand (DASH) scores, and grip strength were measured. Outcomes were measured at early follow-up (6-12 weeks) and at 6-month follow-up. There was a significant improvement in VAS pain score in all treatment groups at early follow-up. DASH score and grip strength improved in the cortisone injection group and the deep friction massage group at early follow-up; these parameters did not improve in the splinting and stretching group. At 6-month follow-up, only patients in the deep friction massage group demonstrated a significant improvement in all outcome measures, including VAS pain score, DASH score, and grip strength. Deep friction massage is an effective treatment for lateral epicondylitis and can be used in patients who have failed other nonoperative treatments, including cortisone injection.

Clinical and patient reported outcomes of bleaching effectiveness.

PubMed

Klaric Sever, Eva; Budimir, Zrinka; Cerovac, Matea; Stambuk, Mario; Par, Matej; Negovetic Vranic, Dubravka; Tarle, Zrinka

2018-01-01

The objective of this study is to evaluate clinical and patient reported outcomes of different bleaching products. Thirty participants were randomly divided into three bleaching groups (n = 10). Bleaching was performed with high concentrations of hydrogen peroxide (HP) - Boost (40%) and Dash (30%), and with prefabricated splints Bite&White (6% HP). Tooth colour was measured before, immediately after, and 1 and 6 months after the bleaching by using classical shade guide and spectrophotometer. Tooth hypersensitivity was self-rated by patients on the Wong-Baker's face scale. Patient satisfaction was evaluated on a 7-point Likert-type scales that measured perceived performance and importance of different characteristics of bleaching treatment. All products were effective in teeth colour change (ΔE > 3.3), which was significantly higher for Boost (p = .016) and Dash (p = .024) than Bite&White treatment. Perception of hypersensitivity was the highest in Boost group, followed by Dash and Bite&White treatment. Most of the patients were satisfied with final tooth colour, length and comfort during treatment, but were dissatisfied with the stability of bleached tooth colour. Materials with the higher concentrations of bleaching agent demonstrated greater bleaching effectiveness than at-home bleaching product, but also a greater hypersensitivity. Lengthening the treatment process, but achieving a more stable tooth colour may improve the perceived value of a bleaching service.

Is posterior synovial plica excision necessary for refractory lateral epicondylitis of the elbow?

PubMed

Rhyou, In Hyeok; Kim, Kang Wook

2013-01-01

Arthroscopic treatments for lateral epicondylitis including débridement of the extensor carpi radialis brevis (ECRB) origin (Baker technique) or resection of the radiocapitellar synovial plica reportedly improve symptoms. However the etiology of the disease and the role of the plica remain unclear. We asked if posterior radiocapitellar synovial plica excision made any additional improvement in pain or function after arthroscopic ECRB release. We retrospectively reviewed 38 patients who had arthroscopic treatment for refractory lateral epicondylitis between November 2003 and October 2009. Twenty patients (Group A) underwent the Baker technique and 18 patients (Group B) underwent a combination of the Baker technique and posterior synovial plica excision. The minimum followup was 36 months (mean, 46 months; range, 36-72 months) for Group A and 25 months (mean, 30 months; range, 25-36 months) for Group B. Postoperatively we obtained VAS pain and DASH scores for each group. Two years postoperatively, we found no differences in the VAS pain score or DASH: the mean VAS pain scores were 0.3 points in Group A and 0.4 points in Group B, and the DASH scores were 5.1 points and 6.1 points respectively. The addition of débridement of the posterior synovial fold did not appear to enhance either pain relief or function compared with the classic Baker technique without decortication.

Functional outcomes after shoulder resection: the patient's perspective.

PubMed

Stevens, Nicole M; Kim, H Mike; Armstrong, April D

2015-09-01

Resection arthroplasty is a salvage procedure used for the treatment of deep-seated infections after total shoulder arthroplasty, hemiarthroplasty, and reverse total shoulder arthroplasty. Previous studies have reported a 50% to 66% rate of pain relief after resection arthroplasty but with significant functional limitations. Our study aimed to qualify the perspective of the patients on their limitations and satisfaction with resection arthroplasty. A retrospective record review of resection arthroplasties performed between September 2003 and December 2012 yielded 14 patients, and 7 completed the survey. The patients completed surveys with the focus on the "patient perspective." Functional scores, including American Shoulder and Elbow Surgeons, Simple Shoulder Test, Disabilities of the Arm, Shoulder, and Hand (DASH), DASH work, and DASH sports, were determined. Pain reduction and functional outcomes were similar to past reports of resection arthroplasty. Five of the 7 patients (71%) reported satisfaction with their resection arthroplasty, and 6 of the 7 patients (86%) would undergo the procedure again if given the choice. Five of the 7 patients (71%) were able to most of activities of daily living. Patients in our study were generally satisfied with their resection arthroplasty. Resection arthroplasty is a reasonable option for treatment of deep-seated periprosthetic infections or for patients with multiple previous failed procedures for total shoulder arthroplasty, hemiarthroplasty. and reverse shoulder arthroplasty. Copyright © 2015 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.

PROMIS Physical Function Computer Adaptive Test Compared With Other Upper Extremity Outcome Measures in the Evaluation of Proximal Humerus Fractures in Patients Older Than 60 Years.

PubMed

Morgan, Jordan H; Kallen, Michael A; Okike, Kanu; Lee, Olivia C; Vrahas, Mark S

2015-06-01

To compare the PROMIS Physical Function Computer Adaptive Test (PROMIS PF CAT) to commonly used traditional PF measures for the evaluation of patients with proximal humerus fractures. Prospective. Two Level I trauma centers. Forty-seven patients older than 60 years with displaced proximal humerus fractures treated between 2006 and 2009. Evaluation included completion of the PROMIS PF CAT, the Constant Shoulder Score, the Disabilities of the Arm, Shoulder, and Hand (DASH) and the Short Musculoskeletal Functional Assessment (SMFA). Observed correlations among the administered PF outcome measures. On average, patients responded to 86 outcome-related items for this study: 4 for the PROMIS PF CAT (range: 4-8 items), 6 for the Constant Shoulder Score, 30 for the DASH, and 46 for the SMFA. Time to complete the PROMIS PF CAT (median completion time = 98 seconds) was significantly less than that for the DASH (median completion time = 336 seconds, P < 0.001) and for the SMFA (median completion time = 482 seconds, P < 0.001). PROMIS PF CAT scores correlated statistically significantly and were of moderate-to-high magnitude with all other PF outcome measure scores administered. This study suggests using the PROMIS PF CAT as a sole PF outcome measure can yield an assessment of upper extremity function similar to those provided by traditional PF measures, while substantially reducing patient assessment time.

Clinimetric evaluation of shoulder disability questionnaires: a systematic review of the literature

PubMed Central

Bot, S; Terwee, C; van der Windt, D A W M; Bouter, L; Dekker, J; de Vet, H C W

2004-01-01

Methods: Systematic literature searches were performed to identify self administered shoulder disability questionnaires. A checklist was developed to evaluate and compare the clinimetric quality of the instruments. Results: Two reviewers identified and evaluated 16 questionnaires by our checklist. Most studies were found for the Disability of the Arm, Shoulder, and Hand scale (DASH), the Shoulder Pain and Disability Index (SPADI), and the American Shoulder and Elbow Surgeons Standardised Shoulder Assessment Form (ASES). None of the questionnaires demonstrated satisfactory results for all properties. Most questionnaires claim to measure several domains (for example, pain, physical, emotional, and social functioning), yet dimensionality was studied in only three instruments. The internal consistency was calculated for seven questionnaires and only one received an adequate rating. Twelve questionnaires received positive ratings for construct validity, although depending on the population studied, four of these questionnaires received poor ratings too. Seven questionnaires were shown to have adequate test-retest reliability (ICC >0.70), but five questionnaires were tested inadequately. In most clinimetric studies only small sample sizes (n<43) were used. Nearly all publications lacked information on the interpretation of scores. Conclusion: The DASH, SPADI, and ASES have been studied most extensively, and yet even published validation studies of these instruments have limitations in study design, sample sizes, or evidence for dimensionality. Overall, the DASH received the best ratings for its clinimetric properties. PMID:15020324

The Brazilian version of the Constant-Murley Score (CMS-BR): convergent and construct validity, internal consistency, and unidimensionality.

PubMed

Barreto, Rodrigo Py Gonçalves; Barbosa, Marcus Levi Lopes; Balbinotti, Marcos Alencar Abaide; Mothes, Fernando Carlos; da Rosa, Luís Henrique Telles; Silva, Marcelo Faria

2016-01-01

To translate and culturally adapt the CMS and assess the validity of the Brazilian version (CMS-BR). The translation was carried out according to the back-translation method by four independent translators. The produced versions were synthesized through extensive analysis and by consensus of an expert committee, reaching a final version used for the cultural adaptation. A field test was conducted with 30 subjects in order to obtain semantic considerations. For the psychometric analyzes, the sample was increased to 110 participants who answered two instruments: CMS-BR and the Disabilities of the Arm, shoulder and Hand (DASH). The CMS-BR and DASH score range from 0 to 100 points. For the first, higher points reflect better function and for the latter, the inverse is true. The validity was verified by Pearson's correlation test, the unidimensionality by factorial analysis, and the internal consistency by Cronbach's alpha. The explained variance was 60.28% with factor loadings ranging from 0.60 to 0.91. The CMS-BR exhibited strong negative correlation with the DASH score (-0.82, p  < 0.05), Cronbach's alpha 0.85, and its total score was strongly correlated with the patient's range of motion (0.93, p  < 0.001). The CMS was satisfactorily adapted for Brazilian Portuguese and demonstrated evidence of validity that allows its use in this population.

Detailed two-dimensional gel proteomic mapping of the feces of the house dust mite Dermatophagoides pteronyssinus and comparison with D. farinae: Reduced trypsin protease content in D. pteronyssinus and different isoforms.

PubMed

Erban, Tomas; Harant, Karel; Hubert, Jan

2017-06-06

Major domestic mite allergens are present in feces. We present a detailed 2D-E-MS/MS proteomic analysis of the Dermatophagoides pteronyssinus feces. Precise cultivation yielded a pure fecal extract. We detected differences in fecal allergens/digestive enzymes between D. pteronyssinus and D. farinae using 2D-E fingerprinting, including unique information on species-specific protease isoforms. Proteomic analysis was performed by 2D-E coupled with MALDI-TOF/TOF identification. The species-specific differences in the fecal extracts of the mites were attributed to trypsin-like proteases known as group 3 allergens. In D. farinae, Der f 3 exhibited high abundance with a pI similar (acidic) to that of the cysteine protease Der f 1 and the chymotrypsin protease Der f 6, whereas in D. pteronyssinus, Der p 3 was rarely detected and exhibited low abundance only at basic pI. Moreover, Der p 9 was detected at a pI of ~ 10, in contrast to Der p 1 and Der p 6, suggesting different compartmentalization in the body. Overall, in D. pteronyssinus feces, allergens of groups 1, 2, 6, and 15 were quantitatively similar to those of D. farinae with the exception of the group 3 and 9 allergens. This work provides novel insights into mite-defecated proteins/digestive enzymes, which are important allergens. Millions of people are affected by allergy and asthma, and their number is growing. In homes, the major triggers of allergy and asthma are the house dust mites Dermatophagoides farinae and D. pteronyssinus, and a clear understanding of the development of diseases caused by these mites is needed. The major sources of mite allergens are their feces, which are deposited in the environment and are easily inhaled as part of aeroplankton. However, descriptions of and comparisons between the major fecal allergens of these two mites are lacking. This study shows that similar group 1 (cysteine protease), 2 (NPC2 family), 6 (chymotrypsin) and 15 (chitinase-like) allergens are present in the feces of these two mite species, as determined by 2D-E mapping, whereas group 3 (trypsin) and 9 (collagenolytic protease) allergens in the feces of the two species are different. The results provide unique MS/MS mapped fingerprints of mite species-specific isoforms in feces. The presence of ubiquitin in mite feces suggests that these proteins participate in the post-translational modification of fecal proteins. The findings are essential for understanding differences between D. farinae and D. pteronyssinus with respect to immunoreactivity, protease activation mechanisms, association with microbes, and food utilization. Copyright © 2017 Elsevier B.V. All rights reserved.

Potential Roles of Protease Inhibitors in Cancer Progression.

PubMed

Yang, Peng; Li, Zhuo-Yu; Li, Han-Qing

2015-01-01

Proteases are important molecules that are involved in many key physiological processes. Protease signaling pathways are strictly controlled, and disorders in protease activity can result in pathological changes such as cardiovascular and inflammatory diseases, cancer and neurological disorders. Many proteases have been associated with increasing tumor metastasis in various human cancers, suggesting important functional roles in the metastatic process because of their ability to degrade the extracellular matrix barrier. Proteases are also capable of cleaving non-extracellular matrix molecules. Inhibitors of proteases to some extent can reduce invasion and metastasis of cancer cells, and slow down cancer progression. In this review, we focus on the role of a few proteases and their inhibitors in tumors as a basis for cancer prognostication and therapy.

Structure of protease-cleaved Escherichia coli α-2-macroglobulin reveals a putative mechanism of conformational activation for protease entrapment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fyfe, Cameron D.; Grinter, Rhys; Josts, Inokentijs

The X-ray structure of protease-cleaved E. coli α-2-macroglobulin is described, which reveals a putative mechanism of activation and conformational change essential for protease inhibition. Bacterial α-2-macroglobulins have been suggested to function in defence as broad-spectrum inhibitors of host proteases that breach the outer membrane. Here, the X-ray structure of protease-cleaved Escherichia coli α-2-macroglobulin is described, which reveals a putative mechanism of activation and conformational change essential for protease inhibition. In this competitive mechanism, protease cleavage of the bait-region domain results in the untethering of an intrinsically disordered region of this domain which disrupts native interdomain interactions that maintain E. colimore » α-2-macroglobulin in the inactivated form. The resulting global conformational change results in entrapment of the protease and activation of the thioester bond that covalently links to the attacking protease. Owing to the similarity in structure and domain architecture of Escherichia coli α-2-macroglobulin and human α-2-macroglobulin, this protease-activation mechanism is likely to operate across the diverse members of this group.« less

Detergent-compatible proteases: microbial production, properties, and stain removal analysis.

PubMed

Niyonzima, Francois Niyongabo; More, Sunil

2015-01-01

Proteases are one of the most important commercial enzymes used in various industrial domains such as detergent and leather industries. The alkaline proteases as well as other detergent-compatible enzymes such as lipases and amylases serve now as the key components in detergent formulations. They break down various stains during fabric washing. The search for detergent-compatible proteases with better properties is a continuous exercise. The current trend is to use detergent-compatible proteases that are stable over a wide temperature range. Although the proteases showing stability at elevated pH have the capacity to be used in detergent formulations, their usage can be significant if they are also stable and compatible with detergent and detergent ingredients, and also able to remove protein stains. Despite the existence of some reviews on alkaline proteases, there is no specification for the use of alkaline proteases as detergent additives. The present review describes the detergent-compatible proteases tested as detergent additives. An overview was provided for screening, optimization, purification, and properties of detergent compatible proteases, with an emphasis on the stability and compatibility of the alkaline proteases with the detergent and detergent compounds, as well as stain removal examination methods.

A New Subtilase-Like Protease Deriving from Fusarium equiseti with High Potential for Industrial Applications.

PubMed

Juntunen, Kari; Mäkinen, Susanna; Isoniemi, Sari; Valtakari, Leena; Pelzer, Alexander; Jänis, Janne; Paloheimo, Marja

2015-09-01

A gene encoding a novel extracellular subtilisin-like protease was cloned from the ascomycete Fusarium equiseti and expressed in Trichoderma reesei. The F. equiseti protease (Fe protease) showed excellent performance in stain removal and good compatibility with several commercial laundry detergent formulations, suggesting that it has high potential for use in various industrial applications. The recombinant enzyme was purified and characterized. The temperature optimum of the Fe protease was 60 °C and it showed high activity in the pH range of 6-10, with a sharp decline in activity at pH above 10. The amino acid specificity of the Fe protease was studied using casein, cytochrome c, and ubiquitin as substrates. The Fe protease had broad substrate specificity: almost all amino acid residues were accepted at position P1, even though it showed some preference for cleavage at the C-terminal side of asparagine and histidine residues. The S4 subsite of Fe protease favors aspartic acid and threonine. The other well-characterized proteases from filamentous fungi, Proteinase K from Engyodontium album, Thermomycolin from Malbranchea sulfurea, and alkaline subtilisins from Bacillus species prefer hydrophobic amino acids in both the S1 and S4 subsites. Due to its different specificity compared to the members of the S8 family of clan SB of proteases, we consider that the Fe protease is a new protease. It does not belong to any previously defined IUBMB groups of proteases.

Structural determinants of tobacco vein mottling virus protease substrate specificity

PubMed Central

Sun, Ping; Austin, Brian P; Tözsér, József; Waugh, David S

2010-01-01

Tobacco vein mottling virus (TVMV) is a member of the Potyviridae, one of the largest families of plant viruses. The TVMV genome is translated into a single large polyprotein that is subsequently processed by three virally encoded proteases. Seven of the nine cleavage events are carried out by the NIa protease. Its homolog from the tobacco etch virus (TEV) is a widely used reagent for the removal of affinity tags from recombinant proteins. Although TVMV protease is a close relative of TEV protease, they exhibit distinct sequence specificities. We report here the crystal structure of a catalytically inactive mutant TVMV protease (K65A/K67A/C151A) in complex with a canonical peptide substrate (Ac-RETVRFQSD) at 1.7-Å resolution. As observed in several crystal structures of TEV protease, the C-terminus (∼20 residues) of TVMV protease is disordered. Unexpectedly, although deleting the disordered residues from TEV protease reduces its catalytic activity by ∼10-fold, an analogous truncation mutant of TVMV protease is significantly more active. Comparison of the structures of TEV and TVMV protease in complex with their respective canonical substrate peptides reveals that the S3 and S4 pockets are mainly responsible for the differing substrate specificities. The structure of TVMV protease suggests that it is less tolerant of variation at the P1′ position than TEV protease. This conjecture was confirmed experimentally by determining kinetic parameters kcat and Km for a series of oligopeptide substrates. Also, as predicted by the cocrystal structure, we confirm that substitutions in the P6 position are more readily tolerated by TVMV than TEV protease. PMID:20862670

Structural determinants of tobacco vein mottling virus protease substrate specificity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, Ping; Austin, Brian P.; Tozer, Jozsef

2010-10-28

Tobacco vein mottling virus (TVMV) is a member of the Potyviridae, one of the largest families of plant viruses. The TVMV genome is translated into a single large polyprotein that is subsequently processed by three virally encoded proteases. Seven of the nine cleavage events are carried out by the NIa protease. Its homolog from the tobacco etch virus (TEV) is a widely used reagent for the removal of affinity tags from recombinant proteins. Although TVMV protease is a close relative of TEV protease, they exhibit distinct sequence specificities. We report here the crystal structure of a catalytically inactive mutant TVMVmore » protease (K65A/K67A/C151A) in complex with a canonical peptide substrate (Ac-RETVRFQSD) at 1.7-{angstrom} resolution. As observed in several crystal structures of TEV protease, the C-terminus ({approx}20 residues) of TVMV protease is disordered. Unexpectedly, although deleting the disordered residues from TEV protease reduces its catalytic activity by {approx}10-fold, an analogous truncation mutant of TVMV protease is significantly more active. Comparison of the structures of TEV and TVMV protease in complex with their respective canonical substrate peptides reveals that the S3 and S4 pockets are mainly responsible for the differing substrate specificities. The structure of TVMV protease suggests that it is less tolerant of variation at the P1{prime} position than TEV protease. This conjecture was confirmed experimentally by determining kinetic parameters k{sub cat} and K{sub m} for a series of oligopeptide substrates. Also, as predicted by the cocrystal structure, we confirm that substitutions in the P6 position are more readily tolerated by TVMV than TEV protease.« less

Synergistic Activity between Two Antifungal Proteins, the Plant Defensin NaD1 and the Bovine Pancreatic Trypsin Inhibitor

PubMed Central

Dawson, Charlotte S.; McKenna, James A.; Quimbar, Pedro; Hayes, Brigitte M. E.; van der Weerden, Nicole L.

2017-01-01

ABSTRACT Defensins are a large family of small, cationic, cysteine-rich proteins that are part of the defense arsenal that plants use for protection against potentially damaging fungal infections. The plant defensin NaD1 from Nicotiana alata is a potent antifungal protein that inhibits growth and kills a variety of fungal pathogens that affect both plant and animal (human) hosts. Some serine protease inhibitors have also been reported to be antifungal molecules, while others have no inhibitory activity against fungi. Here we describe the synergistic activity of the plant defensin NaD1 with a selection of serine protease inhibitors against the plant pathogens Fusarium graminearum and Colletotrichum graminicola and the animal pathogen Candida albicans. The synergistic activity was not related to the protease inhibitory activity of these molecules but may arise from activation of fungal stress response pathways. The bovine pancreatic trypsin inhibitor (BPTI) displayed the most synergy with NaD1. BPTI also acted synergistically with several other antifungal molecules. The observation that NaD1 acts synergistically with protease inhibitors provides the foundation for the design of transgenic plants with improved resistance to fungal disease. It also supports the possibility of naturally occurring accessory factors that function to enhance the activity of innate immunity peptides in biological systems. IMPORTANCE This work describes the increased activity of a natural antifungal peptide in the presence of another antifungal peptide from a different family. This is termed antifungal synergy. Synergy is important for decreasing the amount of antifungal molecule needed to control the disease. Traditionally, naturally occurring antifungal molecules are assayed in isolation. Identification of synergistic interactions between antifungal peptides means that their activities in a complex biological system are likely to be different from what we observe when examining them individually. This study identified synergy between an antifungal peptide and a group of peptides that do not affect fungal growth in vitro. This provides the foundation for generation of transgenic plants with increased resistance to fungal disease and identification of antifungal accessory factors that enhance the activity of innate immune molecules but do not have an antifungal effect on their own. PMID:29062897

Detection of Legume Protease Inhibitors by the Gel-X-ray Film Contact Print Technique

ERIC Educational Resources Information Center

Mulimani, Veerappa H.; Sudheendra, Kulkarni; Giri, Ashok P.

2002-01-01

Redgram (Cajanus cajan L.) extracts have been analyzed for the protease inhibitors using a new, sensitive, simple, and rapid method for detection of electrophoretically separated protease inhibitors. The detection involves equilibrating the gel successively in the protease assay buffer and protease solution, rinsing the gel in assay buffer, and…

Membrane protease degradomics: proteomic identification and quantification of cell surface protease substrates.

PubMed

Butler, Georgina S; Dean, Richard A; Smith, Derek; Overall, Christopher M

2009-01-01

The modification of cell surface proteins by plasma membrane and soluble proteases is important for physiological and pathological processes. Methods to identify shed and soluble substrates are crucial to further define the substrate repertoire, termed the substrate degradome, of individual proteases. Identifying protease substrates is essential to elucidate protease function and involvement in different homeostatic and disease pathways. This characterisation is also crucial for drug target identification and validation, which would then allow the rational design of specific targeted inhibitors for therapeutic intervention. We describe two methods for identifying and quantifying shed cell surface protease targets in cultured cells utilising Isotope-Coded Affinity Tags (ICAT) and Isobaric Tags for Relative and Absolute Quantification (iTRAQ). As a model system to develop these techniques, we chose a cell-membrane expressed matrix metalloproteinase, MMP-14, but the concepts can be applied to proteases of other classes. By over-expression, or conversely inhibition, of a particular protease with careful selection of control conditions (e.g. vector or inactive protease) and differential labelling, shed proteins can be identified and quantified by mass spectrometry (MS), MS/MS fragmentation and database searching.

Tripeptide inhibitors of dengue and West Nile virus NS2B-NS3 protease.

PubMed

Schüller, Andreas; Yin, Zheng; Brian Chia, C S; Doan, Danny N P; Kim, Hyeong-Kyu; Shang, Luqing; Loh, Teck Peng; Hill, Jeffery; Vasudevan, Subhash G

2011-10-01

A series of tripeptide aldehyde inhibitors were synthesized and their inhibitory effect against dengue virus type 2 (DENV2) and West Nile virus (WNV) NS3 protease was evaluated side by side with the aim to discover potent flaviviral protease inhibitors and to examine differences in specificity of the two proteases. The synthesized inhibitors feature a varied N-terminal cap group and side chain modifications of a P2-lysine residue. In general a much stronger inhibitory effect of the tripeptide inhibitors was observed toward WNV protease. The inhibitory concentrations against DENV2 protease were in the micromolar range while they were submicromolar against WNV. The data suggest that a P2-arginine shifts the specificity toward DENV2 protease while WNV protease favors a lysine in the P2 position. Peptides with an extended P2-lysine failed to inhibit DENV2 protease suggesting a size-constrained S2 pocket. Our results generally encourage the investigation of di- and tripeptide aldehydes as inhibitors of DENV and WNV protease. Copyright © 2011 Elsevier B.V. All rights reserved.

2-D zymographic analysis of Broccoli (Brassica oleracea L. var. Italica) florets proteases: follow up of cysteine protease isotypes in the course of post-harvest senescence.

PubMed

Rossano, Rocco; Larocca, Marilena; Riccio, Paolo

2011-09-01

Zymographic analysis of Broccoli florets (Brassica oleracea L. var. Italica) revealed the presence of acidic metallo-proteases, serine proteases and cysteine proteases. Under conditions which were denaturing for the other proteases, the study was restricted to cysteine proteases. 2-D zymography, a technique that combines IEF and zymography was used to show the presence of 11 different cysteine protease spots with molecular mass of 44 and 47-48kDa and pIs ranging between 4.1 and 4.7. pI differences could be ascribed to different degrees of phosphorylation that partly disappeared in the presence of alkaline phosphatase. Post-harvest senescence of Broccoli florets was characterized by decrease in protein and chlorophyll contents and increase of protease activity. In particular, as determined by 2-D zymography, the presence of cysteine protease clearly increased during senescence, a finding that may represent a useful tool for the control of the aging process. Copyright © 2011 Elsevier GmbH. All rights reserved.

Protease resistance of infectious prions is suppressed by removal of a single atom in the cellular prion protein.

PubMed

Leske, Henning; Hornemann, Simone; Herrmann, Uli Simon; Zhu, Caihong; Dametto, Paolo; Li, Bei; Laferriere, Florent; Polymenidou, Magdalini; Pelczar, Pawel; Reimann, Regina Rose; Schwarz, Petra; Rushing, Elisabeth Jane; Wüthrich, Kurt; Aguzzi, Adriano

2017-01-01

Resistance to proteolytic digestion has long been considered a defining trait of prions in tissues of organisms suffering from transmissible spongiform encephalopathies. Detection of proteinase K-resistant prion protein (PrPSc) still represents the diagnostic gold standard for prion diseases in humans, sheep and cattle. However, it has become increasingly apparent that the accumulation of PrPSc does not always accompany prion infections: high titers of prion infectivity can be reached also in the absence of protease resistant PrPSc. Here, we describe a structural basis for the phenomenon of protease-sensitive prion infectivity. We studied the effect on proteinase K (PK) resistance of the amino acid substitution Y169F, which removes a single oxygen atom from the β2-α2 loop of the cellular prion protein (PrPC). When infected with RML or the 263K strain of prions, transgenic mice lacking wild-type (wt) PrPC but expressing MoPrP169F generated prion infectivity at levels comparable to wt mice. The newly generated MoPrP169F prions were biologically indistinguishable from those recovered from prion-infected wt mice, and elicited similar pathologies in vivo. Surprisingly, MoPrP169F prions showed greatly reduced PK resistance and density gradient analyses showed a significant reduction in high-density aggregates. Passage of MoPrP169F prions into mice expressing wt MoPrP led to full recovery of protease resistance, indicating that no strain shift had taken place. We conclude that a subtle structural variation in the β2-α2 loop of PrPC affects the sensitivity of PrPSc to protease but does not impact prion replication and infectivity. With these findings a specific structural feature of PrPC can be linked to a physicochemical property of the corresponding PrPSc.

Inhibitors of serine proteases decrease sperm penetration during porcine fertilization in vitro by inhibiting sperm binding to the zona pellucida and acrosome reaction.

PubMed

Beek, J; Nauwynck, H; Appeltant, R; Maes, D; Van Soom, A

2015-11-01

Serine proteases are involved in mammalian fertilization. Inhibitors of serine proteases can be applied to investigate at which point these enzymes exert their action. We selected two serine protease inhibitors, 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF, 100 μM) and soybean trypsin inhibitor (STI, 5 μM) from Glycine max, via previous dose-response IVF experiments and sperm toxicity tests. In the present study, we evaluated how these inhibitors affect porcine fertilization in vitro as calculated on total fertilization rate, polyspermy rate, and the sperm number per fertilized oocyte of cumulus-intact, cumulus-free, and zona-free oocytes. In the control group (no inhibitor), these parameters were 86%, 49%, and 2.2 for cumulus-intact oocytes and 77%, 43%, and 2.2 for cumulus-free oocytes (6-hour gamete incubation period, 1.25 × 10(5) spermatozoa/mL). 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride and STI significantly reduced total fertilization and polyspermy rate in cumulus-intact and cumulus-free oocytes (P < 0.05). Total fertilization rates were respectively 65% and 53% (AEBSF) and 36% and 17% (STI). Inhibition rates were higher in cumulus-free oocytes than in cumulus-intact oocytes, indicating that inhibitors exerted their action after sperm passage through the cumulus. 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride but not STI reduced sperm binding to the ZP. The acrosome reaction was significantly inhibited by both inhibitors. Only 40.4% (AEBSF) and 11.4% (STI) of spermatozoa completed a calcium-induced acrosome reaction compared to 86.7% of spermatozoa in the control group. There was no effect on sperm binding or fertilization parameters in zona-free oocytes. In conclusion, sperm-zona binding and acrosome reaction were inhibited by serine protease inhibitors during porcine IVF. Copyright © 2015 Elsevier Inc. All rights reserved.

Proteases Revisited: Roles and Therapeutic Implications in Fibrosis

PubMed Central

Kryczka, Jakub

2017-01-01

Proteases target many substrates, triggering changes in distinct biological processes correlated with cell migration, EMT/EndMT and fibrosis. Extracellular protease activity, demonstrated by secreted and membrane-bound protease forms, leads to ECM degradation, activation of other proteases (i.e., proteolysis of nonactive zymogens), decomposition of cell-cell junctions, release of sequestered growth factors (TGF-β and VEGF), activation of signal proteins and receptors, degradation of inflammatory inhibitors or inflammation-related proteins, and changes in cell mechanosensing and motility. Intracellular proteases, mainly caspases and cathepsins, modulate lysosome activity and signal transduction pathways. Herein, we discuss the current knowledge on the multidimensional impact of proteases on the development of fibrosis. PMID:28642633

Purification and Characterization of a Protease Produced by a Planomicrobium sp. L-2 from Gut of Octopus vulgaris

PubMed Central

Liu, Qing; Sun, Shujing; Piao, Meizi; Yang, Ji Young

2013-01-01

Protease widely exists in the digestive tract of animals and humans, playing a very important role in protein digestion and absorption. In this study, a high protease-producing strain Planomicrobium sp. L-2 was isolated and identified from the digestive tract of Octopus variabilis. The strain was identified by physiological and biochemical experiments and 16S rDNA sequences analysis. A protease was obtained from the strain Planomicrobium sp. L-2 through ammonium sulfate precipitation, dialysis and enrichment, DEAE-Sephadex A50 anion-exchange chromatography, and Sephadex G-100 gel chromatography. The molecular weight and properties of the protease were characterized, including optimum temperature and pH, thermal stability, protease inhibitions and metal ions. According to our results, the protease from Planomicrobium sp. L-2 strain designated as F1-1 was obtained by three-step separation and purification from crude enzyme. The molecular weight of the protease was 61.4 kDa and its optimum temperature was 40°C. The protease F1-1 showed a broad pH profile for casein hydrolysis between 5.0~11.0. No residual activity was observed after incubation for 40 min at 60°C and 60 min at 50°C. F1-1 protease was inhibited by Mn2+, Hg2+, Pb2+, Zn2+, and Cu2+ ions, as well as PMSF, indicating that the protease F1-1 was a serine protease. Additionally, research basis provided by this study could be considered for industrial application of octopus intestinal proteases. PMID:24551830

Purification and characterisation of a salt-stable protease from the halophilic archaeon Halogranum rubrum.

PubMed

Gao, Ruichang; Shi, Tong; Liu, Xiangdong; Zhao, Mengqin; Cui, Henglin; Yuan, Li

2017-03-01

Because proteases play an important role in the fermentation of fish sauce, the purification and characterisation of an extracellular protease from the halophilic archaeon Halogranum rubrum was investigated. The molecular mass of the protease was estimated to be approximately 47 kDa based on sodium dodecyl sulfate-polyacrylamide gel electropheresis (SDS-PAGE) and native-PAGE analysis. The optimum conditions for catalytic activity were pH 8.0 and 50°C. The protease showed alkaline stability (pH 7.0-10.0). The protease also exhibited novel catalytic ability over a broad range of salinity (NaCl 0-3 mol L -1 ). Calcium ion enhanced the proteolytic activity of the enzyme. The K m and V max values of the purified protease for casein were calculated to be 4.89 mg mL -1 and 1111.11 U mL -1 , respectively. The protease was strongly inhibited by ethylenediamine tetraacetic acid (EDTA) and phenylmethanesulfonyl fluoride (PMSF). Meanwhile, the protease was stable in the presence of Triton X-100, isopropanol, ethanol or dithio-bis-nitrobenzoic (DTNB), but was inhibited by sodium dodecyl sulfate (SDS), dimethyl sulfoxide (DMSO) or methanol. MALDI -TOF/TOF MS analysis revealed that the protease shared some functional traits with protease produced by Halogranum salarium. Furthermore, it exhibited high hydrolytic activity on silver carp myosin protein. The protease is an alkaline and salt-tolerant enzyme that hydrolyses silver carp myosin with high efficiency. These excellent characteristics make this protease an attractive candidate for industrial use in low-salt fish sauce fermentation. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Recombinant expression and antigenic properties of a 32-kilodalton extracellular alkaline protease, representing a possible virulence factor from Aspergillus fumigatus.

PubMed Central

Moser, M; Menz, G; Blaser, K; Crameri, R

1994-01-01

A 32-kDa nonglycosylated alkaline protease (EC 3.4.1.14) with elastolytic activity, secreted by the opportunistic pathogen Aspergillus fumigatus ATCC 42202, is suggested to be a virulence factor of this fungus. The enzyme is a serine protease of the subtilisin family, and its cDNA nucleotide sequence has recently been reported. We have cloned the cDNA encoding the mature protease into a high-level Escherichia coli expression plasmid and produced the recombinant protease as a fusion protein with a six-adjacent-histidine affinity tag at the carboxy terminus. Subsequently, the recombinant protease was purified to homogeneity, with affinity chromatography yielding 30 to 40 mg of recombinant protease per liter of E. coli culture. Refolded recombinant protease, in comparison with native protease, demonstrated weak enzymatic activity but similar immunochemical characteristics as analyzed by antigen-specific enzyme-linked immunosorbent assay (ELISA), competition ELISA, and immunoblotting assays. To assess the allergenic potential of the protease, sera from patients with allergic bronchopulmonary aspergillosis and sera from healthy control individuals were analyzed by ELISA and immunoblotting techniques. Sera from patients with allergic bronchopulmonary aspergillosis did not have protease-specific immunoglobulin E (IgE) antibodies and, remarkably, did not show significantly elevated protease-specific IgG antibody levels compared with those in sera from healthy control individuals. This suggests that the alkaline protease from A. fumigatus does not elicit IgE antibodies and has weak immunogenicity, a property which may explain fungus persistence in allergic individuals. Images PMID:8112866

Optimum production and characterization of an acid protease from marine yeast Metschnikowia reukaufii W6b

NASA Astrophysics Data System (ADS)

Li, Jing; Peng, Ying; Wang, Xianghong; Chi, Zhenming

2010-12-01

The marine yeast strain W6b isolated from sediment of the South China Sea was found to produce a cell-bound acid protease. The crude acid protease produced by this marine yeast showed the highest activity at pH 3.5 and 40 °C. The optimal pH and temperature for the crude acid protease were in agreement with those for acid protease produced by the terrestrial yeasts. The optimal medium of the acid protease production was seawater containing 1.0% glucose, 1.5% casein, and 0.5% yeast extract, and the optimal cultivation conditions of the acid protease production were pH 4.0, a temperature of 25 °C and a shaking speed of 140 rmin-1. Under the optimal conditions, 72.5 UmL-1 of acid protease activity could be obtained in cell suspension within 48 h of fermentation at shake flask level. The acid protease production was induced by high-molecular-weight nitrogen sources and repressed by low-molecular-weight nitrogen sources. Skimmed-milk-clotting test showed that the crude acid protease from the cell suspension of the yeast W6b had high skimmed milk coagulability. The acid protease produced by M. reukaufii W6b may have highly potential applications in cheese, food and fermentation industries.

Enteric bacterial proteases in inflammatory bowel disease- pathophysiology and clinical implications

PubMed Central

Carroll, Ian M; Maharshak, Nitsan

2013-01-01

Numerous reports have identified a dysbiosis in the intestinal microbiota in patients suffering from inflammatory bowel diseases (IBD), yet the mechanism(s) in which this complex microbial community initiates or perpetuates inflammation remains unclear. The purpose of this review is to present evidence for one such mechanism that implicates enteric microbial derived proteases in the pathogenesis of IBD. We highlight and discuss studies demonstrating that proteases and protease receptors are abundant in the digestive system. Additionally, we investigate studies demonstrating an association between increased luminal protease activity and activation of protease receptors, ultimately resulting in increased intestinal permeability and exacerbation of colitis in animal models as well as in human IBD. Proteases are essential for the normal functioning of bacteria and in some cases can serve as virulence factors for pathogenic bacteria. Although not classified as traditional virulence factors, proteases originating from commensal enteric bacteria also have a potential association with intestinal inflammation via increased enteric permeability. Reports of increased protease activity in stools from IBD patients support a possible mechanism for a dysbiotic enteric microbiota in IBD. A better understanding of these pathways and characterization of the enteric bacteria involved, their proteases, and protease receptors may pave the way for new therapeutic approaches for these diseases. PMID:24431894

Single-arm open-label study of Durolane (NASHA nonanimal hyaluronic acid) for the treatment of osteoarthritis of the thumb.

PubMed

Velasco, Eloisa; Ribera, Mª Victoria; Pi, Joan

2017-01-01

Osteoarthritis of the trapeziometacarpal (TMC) joint of the thumb - also known as rhizarthrosis - is painful and has a significant impact on quality of life. Intra-articular injection of hyaluronic acid may potentially meet the need for effective, minimally invasive intervention in patients not responding adequately to initial treatment. We aimed to confirm the safety and effectiveness of viscosupplementation with Durolane (NASHA nonanimal hyaluronic acid) in rhizarthrosis. This was a prospective, single-arm, multicenter, open-label study with a 6-month follow-up period. Eligible patients had Eaton-Littler grade II-III rhizarthrosis in one TMC joint with pain and visual analog scale (VAS) pain score ≥4 (scale: 0-10). A single injection of NASHA was administered to the affected TMC joint. The primary effectiveness variable was change from baseline in VAS pain score. Thirty-five patients (mean age 60.8 years; 85.7% female) received NASHA and completed the study. The least-squares mean change from baseline in VAS pain score over 6 months was -2.00, a reduction of 27.8% ( p <0.001). The reduction in pain exceeded 25% as early as month 1 (26.5%), and gradual improvement was observed throughout the 6-month follow-up period. Secondary effectiveness parameters included QuickDASH (shortened version of Disabilities of the Arm, Shoulder, and Hand [DASH]), Kapandji thumb opposition test, radial abduction, metacarpophalangeal (MCP) joint flexion, and pinch (clamp) strength. Most of these measurements showed statistically significant improvements from baseline over 6 months. Five adverse events (injection site reactions) were reported in four patients (11.4%), and there were no serious or allergic reactions. This study suggests that viscosupplementation using NASHA is effective and well tolerated in treating the symptoms of rhizarthrosis.

Percutaneous ultrasonic tenotomy for chronic elbow tendinosis: a prospective study.

PubMed

Barnes, Darryl E; Beckley, James M; Smith, Jay

2015-01-01

Elbow tendinopathy is the most common cause of elbow pain affecting active populations. Surgical excision is reserved for patients with refractory symptoms. Percutaneous ultrasonic tenotomy performed under local anesthesia also removes degenerated tissue and therefore provides an alternative treatment option to surgical excision. This investigation prospectively documented the safety and 1-year efficacy of ultrasonic percutaneous tenotomy performed by a single operator. Nineteen patients, aged 38 to 67 years, in whom >6 months of conservative management for medial (7) or lateral (12) elbow tendinopathy had failed were prospectively studied. All patients were treated with percutaneous ultrasonic tenotomy of the elbow by a single operator. Visual analog scale (VAS) for pain, the 11-item version of the Disabilities of the Arm, Shoulder, and Hand (Quick DASH) index, and the Mayo Elbow Performance Score (MEPS) were assessed by an independent observer before treatment and at 6 weeks, 3 months, 6 months, and 12 months after treatment. No procedural complications occurred. Total treatment time was <15 minutes, and ultrasonic energy time averaged 38.6 ± 8.8 seconds per procedure. Average VAS scores were significantly improved from 6.4 to 2.6 at 6 weeks and were 0.7 at 12 months (P < .0001). Similar improvement occurred with the Quick DASH (pretreatment, 44.1; 12 months, 8.6, P < .0001) and MEPS (pretreatment, 59.1; 12 months, 83.4; P < .0001). Percutaneous ultrasonic tenotomy performed under local anesthesia appears to be a safe and effective treatment option for chronic, refractory lateral or medial elbow tendinopathy up to 1 year after the procedure. Copyright © 2015 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.

Carrier diffusion as a measure of carrier/exciton transfer rate in InAs/InGaAsP/InP hybrid quantum dot-quantum well structures emitting at telecom spectral range

NASA Astrophysics Data System (ADS)

Rudno-Rudziński, W.; Biegańska, D.; Misiewicz, J.; Lelarge, F.; Rousseau, B.; Sek, G.

2018-01-01

We investigate the diffusion of photo-generated carriers (excitons) in hybrid two dimensional-zero dimensional tunnel injection structures, based on strongly elongated InAs quantum dots (called quantum dashes, QDashes) of various heights, designed for emission at around 1.5 μm, separated by a 3.5 nm wide barrier from an 8 nm wide In0.64Ga0.36As0.78P0.22 quantum well (QW). By measuring the spectrally filtered real space images of the photoluminescence patterns with high resolution, we probe the spatial extent of the emission from QDashes. Deconvolution with the exciting light spot shape allows us to extract the carrier/exciton diffusion lengths. For the non-resonant excitation case, the diffusion length depends strongly on excitation power, pointing at carrier interactions and phonons as its main driving mechanisms. For the case of excitation resonant with absorption in the adjacent QW, the diffusion length does not depend on excitation power for low excitation levels since the generated carriers do not have sufficient excess kinetic energy. It is also found that the diffusion length depends on the quantum-mechanical coupling strength between QW and QDashes, controlled by changing the dash size. It influences the energy difference between the QDash ground state of the system and the quantum well levels, which affects the tunneling rates. When that QW-QDash level separation decreases, the probability of capturing excitons generated in the QW by QDashes increases, which is reflected by the decreased diffusion length from approx. 5 down to 3 μm.

Effectiveness of commercial inhibitors against subtype F HIV-1 protease.

PubMed

Krauchenco, Sandra; Martins, Nadia H; Sanches, Mario; Polikarpov, Igor

2009-06-01

Subtype F wild type HIV protease has been kinetically characterized using six commercial inhibitors (amprenavir, indinavir, lopinavir, nelfinavir, ritonavir and saquinavir) commonly used for HIV/AIDS treatment, as well as inhibitor TL-3 and acetyl-pepstatin. We also obtained kinetic parameters for two multi-resistant proteases (one of subtype B and one of subtype F) harboring primary and secondary mutations selected by intensive treatment with ritonavir/nelfinavir. This newly obtained biochemical data shows that all six studied commercially available protease inhibitors are significantly less effective against subtype F HIV proteases than against HIV proteases of subtype B, as judged by increased K(i) and biochemical fitness (vitality) values. Comparison with previously reported kinetic values for subtype A and C HIV proteases show that subtype F wild type proteases are significantly less susceptible to inhibition. These results demonstrate that the accumulation of natural polymorphisms in subtype F proteases yields catalytically more active enzymes with a large degree of cross-resistance, which thus results in strong virus viability.

Aspartic protease inhibitors as potential anti-Candida albicans drugs: impacts on fungal biology, virulence and pathogenesis.

PubMed

Braga-Silva, L A; Santos, A L S

2011-01-01

Mycoses are still one of the most problematic illnesses worldwide, especially affecting immunocompromised individuals. The development of novel antifungal drugs is becoming more demanding every day, since existing drugs either have too many side effects or they tend to lose effectiveness due to the resistant fungal strains. In this scenario, Candida albicans is still the main fungal pathogen isolated in hospitals. Pathogenicity results essentially from modifications of the host defense mechanisms that secondarily initiate transformations in the fungal behavior. The pathogenesis of C. albicans is multifactorial and different virulence attributes are important during the various stages of infection. Some virulence factors, like the secreted aspartic proteases (Saps), play a role in several infection stages and the inhibition of one of the many stages may contribute to the containment of the pathogen and thus should help in the treatment of disease. Therefore, Saps are potential targets for the development of novel anti-C. albicans drugs. Herein, we review the beneficial properties of pepstatin A and aspartic-type protease inhibitors used in the anti-human immunodeficiency virus chemotherapy on C. albicans, with particular emphasis in the effects on Sap activity, proliferation, morphogenesis (yeasts into mycelia transformation), ultrastructural architecture, adhesion to mammalian cells and abiotic materials, modulation of unrelated virulence factors (e.g., surface glycoconjugates, lipases and sterol), experimental candidiasis infection as well as synergistic properties when conjugated with classical antifungals. Collectively, these positive findings have stimulated the search for novel natural and/or synthetic pharmacological compounds with anti-aspartic protease properties against the human opportunistic fungus C. albicans.

Protease-mediated Inflammation: An In Vitro Human Keratinocyte-based Screening Tool for Anti-inflammatory Drug Nanocarrier Systems

NASA Astrophysics Data System (ADS)

Frombach, Janna; Lohan, Silke B.; Lemm, Davina; Gruner, Paul; Hasler, Julia; Ahlberg, Sebastian; Blume-Peytavi, Ulrike; Unbehauen, Michael; Haag, Rainer; Meinke, Martina C.; Vogt, Annika

2018-05-01

Refined encapsulation approaches in dermatotherapy gain increased interest. There is need of reproducible in vitro systems representing disease features to screen drug delivery systems for preclinical assessment. Inflammatory human skin diseases are commonly accompanied by abnormal epidermal differentiation and barrier impairment. Serine proteases (SPs) and their inhibitors play a critical role in such dysfunctional differentiation. SPs also initiate cellular pathways via activation of protease-activated receptors, which contribute to inflammation. Thus, function and activity of SPs should be considered for the design of new therapies of such disorders. Herein, we established a novel simplified cell culture model, based on SP-mediated inflammation suitable to assess nanocarriers loaded with anti-inflammatory drugs. SP-mediated inflammation and the regulatory effect of free or encapsulated dexamethasone were determined by measuring interleukin-6 and interleukin-8 in culture medium of HaCaT (human adult low calcium temperature)-keratinocytes. Additionally, radical formation was analyzed by electron paramagnetic resonance spectroscopy. Cellular uptake of core-multishell nanocarriers was investigated by fluorescence microscopy. Cytotoxicity of all additives was determined by a viability assay. SP-Stimulation of keratinocytes resulted in increased radical production and release of inflammatory cytokines without affecting cell viability. Induced inflammation was successfully downregulated by addition of free or encapsulated dexamethasone. SP-addition can be used as inflammatory stimulus in cell culture to mimic effects of aberrant enzymatic activities found in skin of atopic dermatitis patients. The set-up is appropriate as a preliminary test to examine the effectiveness of new molecules or delivery-systems to counteract serine protease-mediated inflammatory processes prior to skin studies.

Suppression of calpain expression by NSAIDs is associated with inhibition of cell migration in rat duodenum.

PubMed

Silver, Kristopher; Littlejohn, A; Thomas, Laurel; Bawa, Bhupinder; Lillich, James D

2017-05-15

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used for the alleviation of pain and inflammation, but these drugs are also associated with a suite of negative side effects. Gastrointestinal (GI) toxicity is particularly concerning since it affects an estimated 70% of individuals taking NSAIDs routinely, and evidence suggests the majority of toxicity is occurring in the small intestine. Traditionally, NSAID-induced GI toxicity has been associated with indiscriminate inhibition of cyclooxygenase isoforms, but other mechanisms, including inhibition of cell migration, intestinal restitution, and wound healing, are likely to contribute to toxicity. Previous efforts demonstrated that treatment of cultured intestinal epithelial cells (IEC) with NSAIDs inhibits expression and activity of calpain proteases, but the effects of specific inhibition of calpain expression in vitro or the effects of NSAIDs on intestinal cell migration in vivo remain to be determined. Accordingly, we examined the effect of suppression of calpain protease expression with siRNA on cell migration in cultured IECs and evaluated the effects of NSAID treatment on epithelial cell migration and calpain protease expression in rat duodenum. Our results show that calpain siRNA inhibits protease expression and slows migration in cultured IECs. Additionally, NSAID treatment of rats slowed migration up the villus axis and suppressed calpain expression in duodenal epithelial cells. Our results are supportive of the hypothesis that suppression of calpain expression leading to slowing of cell migration is a potential mechanism through which NSAIDs cause GI toxicity. Copyright © 2017 Elsevier B.V. All rights reserved.

Deletion of Cysteine Cathepsins B or L Yields Differential Impacts on Murine Skin Proteome and Degradome*

PubMed Central

Tholen, Stefan; Biniossek, Martin L.; Gansz, Martina; Gomez-Auli, Alejandro; Bengsch, Fee; Noel, Agnes; Kizhakkedathu, Jayachandran N.; Boerries, Melanie; Busch, Hauke; Reinheckel, Thomas; Schilling, Oliver

2013-01-01

Numerous studies highlight the fact that concerted proteolysis is essential for skin morphology and function. The cysteine protease cathepsin L (Ctsl) has been implicated in epidermal proliferation and desquamation, as well as in hair cycle regulation. In stark contrast, mice deficient in cathepsin B (Ctsb) do not display an overt skin phenotype. To understand the systematic consequences of deleting Ctsb or Ctsl, we determined the protein abundances of >1300 proteins and proteolytic cleavage events in skin samples of wild-type, Ctsb−/−, and Ctsl−/− mice via mass-spectrometry-based proteomics. Both protease deficiencies revealed distinct quantitative changes in proteome composition. Ctsl−/− skin revealed increased levels of the cysteine protease inhibitors cystatin B and cystatin M/E, increased cathepsin D, and an accumulation of the extracellular glycoprotein periostin. Immunohistochemistry located periostin predominantly in the hypodermal connective tissue of Ctsl−/− skin. The proteomic identification of proteolytic cleavage sites within skin proteins revealed numerous processing sites that are underrepresented in Ctsl−/− or Ctsb−/− samples. Notably, few of the affected cleavage sites shared the canonical Ctsl or Ctsb specificity, providing further evidence of a complex proteolytic network in the skin. Novel processing sites in proteins such as dermokine and Notch-1 were detected. Simultaneous analysis of acetylated protein N termini showed prototypical mammalian N-alpha acetylation. These results illustrate an influence of both Ctsb and Ctsl on the murine skin proteome and degradome, with the phenotypic consequences of the absence of either protease differing considerably. PMID:23233448

Purification and crystallization of dengue and West Nile virus NS2B–NS3 complexes

DOE Office of Scientific and Technical Information (OSTI.GOV)

D’Arcy, Allan, E-mail: allan.darcy@novartis.com; Chaillet, Maxime; Schiering, Nikolaus

Crystals of dengue serotype 2 and West Nile virus NS2B–NS3 protease complexes have been obtained and the crystals of both diffract to useful resolution. Sample homogeneity was essential for obtaining X-ray-quality crystals of the dengue protease. Controlled proteolysis produced a crystallizable fragment of the apo West Nile virus NS2B–NS3 and crystals were also obtained in the presence of a peptidic inhibitor. Both dengue and West Nile virus infections are an increasing risk to humans, not only in tropical and subtropical areas, but also in North America and parts of Europe. These viral infections are generally transmitted by mosquitoes, but maymore » also be tick-borne. Infection usually results in mild flu-like symptoms, but can also cause encephalitis and fatalities. Approximately 2799 severe West Nile virus cases were reported this year in the United States, resulting in 102 fatalities. With this alarming increase in the number of West Nile virus infections in western countries and the fact that dengue virus already affects millions of people per year in tropical and subtropical climates, there is a real need for effective medicines. A possible therapeutic target to combat these viruses is the protease, which is essential for virus replication. In order to provide structural information to help to guide a lead identification and optimization program, crystallizations of the NS2B–NS3 protease complexes from both dengue and West Nile viruses have been initiated. Crystals that diffract to high resolution, suitable for three-dimensional structure determinations, have been obtained.« less

Antiacanthain A: New proteases isolated from Bromelia antiacantha Bertol. (Bromeliaceae).

PubMed

Vallés, Diego; Cantera, Ana M B

2018-07-01

Crude extract (CE) from pulp of Bromelia antiacantha Bertol. mature fruit, contains at least 3 cysteine proteases with proteolytic activity. By single step cation exchange chromatography (Hi-trap SP-HP) of partially purified CE, the protease with the lowest pI, Antiacanthain A (AntA), was isolated. It showed maximum activity at pH9, and 75% of remaining activity was maintained over a wide pH range (pH6-10). The AntA activity exhibits a constant increase up to 70°C. Maintains almost 100% of its activity at 45 at pH6 and 9. A 60% of AntA was active by titration with specific inhibitor, E64. Amidasic activity was studied with pyroglutamyl-phenyl-leucyl-paranitroaniline (PFLNA) substrate having higher AntA catalytic efficiency of (k cat /K m =470s -1 M -1 ) relative to stem bromelain (k cat /K m =305s -1 M -1 ). Esterase activity using p-nitrophenyl esters of N-α-CBZ-l-Lysine (z-L-LysONp) showed a 10-fold higher catalytic efficiency for AntA (k cat /K m =6376s -1 M -1 ) relative to stem bromelain (k cat /K m =688s -1 M -1 ). Incubation with 8M Urea did not affect AntA activity and remained unchanged for 18h, with 6M GndHCl resulted in a 41% decrease in activity after 30min incubation, maintained this activity 18h. AntA exhibits high sequence identity with proteases of the Bromeliaceae family. Copyright © 2018 Elsevier B.V. All rights reserved.

Development of marine biotechnology as a resource for novel proteases and their role in modern biotechnology.

PubMed

Homaei, Ahmad; Lavajoo, Fatemeh; Sariri, Reyhaneh

2016-07-01

Marine environment consists of the largest sources diversified genetic pool of material with an enormous potential for a wide variety of enzymes including proteases. A protease hydrolyzes the peptide bond and most of proteases possess many industrial applications. Marine proteases differ considerably from those found in internal or external organs of invertebrates and vertebrates. In common with all enzymes, external factors such as temperature, pH and type of media are important for the activity, catalytic efficiency, stability and proper functioning of proteases. In this review valuable characteristics of proteases in marine organisms and their applications are gathered from a wide literature survey. Considering their biochemical significance and their increasing importance in biotechnology, a thorough understanding of marine proteases functioning could be of prime importance. Copyright © 2016 Elsevier B.V. All rights reserved.

Proteases in Escherichia coli and Staphylococcus aureus confer reduced susceptibility to lactoferricin B.

PubMed

Ulvatne, Hilde; Haukland, Hanne Husom; Samuelsen, Ørjan; Krämer, Manuela; Vorland, Lars H

2002-10-01

Lactoferricin B is a cationic antimicrobial peptide derived from the N-terminal part of bovine lactoferrin. The effect of bacterial proteases on the antibacterial activity of lactoferricin B towards Escherichia coli and Staphylococcus aureus was investigated using various protease inhibitors and protease-deficient E. coli mutants. Sodium-EDTA, a metalloprotease inhibitor, was the most efficient inhibitors in both species, but combinations of sodium-EDTA with other types of protease inhibitor gave a synergic effect. The results indicate that several groups of proteases are involved in resistance to lactoferricin B in both E. coli and S. aureus. We also report that genetic inactivation of the heat shock-induced serine protease DegP increased the susceptibility to lactoferricin B in E. coli, suggesting that this protease, at least, is involved in reduced susceptibility to lactoferricin B.

Viral evolution in response to the broad-based retroviral protease inhibitor TL-3.

PubMed

Bühler, B; Lin, Y C; Morris, G; Olson, A J; Wong, C H; Richman, D D; Elder, J H; Torbett, B E

2001-10-01

TL-3 is a protease inhibitor developed using the feline immunodeficiency virus protease as a model. It has been shown to efficiently inhibit replication of human, simian, and feline immunodeficiency viruses and therefore has broad-based activity. We now demonstrate that TL-3 efficiently inhibits the replication of 6 of 12 isolates with confirmed resistance mutations to known protease inhibitors. To dissect the spectrum of molecular changes in protease and viral properties associated with resistance to TL-3, a panel of chronological in vitro escape variants was generated. We have virologically and biochemically characterized mutants with one (V82A), three (M46I/F53L/V82A), or six (L24I/M46I/F53L/L63P/V77I/V82A) changes in the protease and structurally modeled the protease mutant containing six changes. Virus containing six changes was found to be 17-fold more resistant to TL-3 in cell culture than was wild-type virus but maintained similar in vitro replication kinetics compared to the wild-type virus. Analyses of enzyme activity of protease variants with one, three, and six changes indicated that these enzymes, compared to wild-type protease, retained 40, 47, and 61% activity, respectively. These results suggest that deficient protease enzymatic activity is sufficient for function, and the observed protease restoration might imply a selective advantage, at least in vitro, for increased protease activity.

Viral Evolution in Response to the Broad-Based Retroviral Protease Inhibitor TL-3†

PubMed Central

Bühler, Bernd; Lin, Ying-Chuan; Morris, Garrett; Olson, Arthur J.; Wong, Chi-Huey; Richman, Douglas D.; Elder, John H.; Torbett, Bruce E.

2001-01-01

TL-3 is a protease inhibitor developed using the feline immunodeficiency virus protease as a model. It has been shown to efficiently inhibit replication of human, simian, and feline immunodeficiency viruses and therefore has broad-based activity. We now demonstrate that TL-3 efficiently inhibits the replication of 6 of 12 isolates with confirmed resistance mutations to known protease inhibitors. To dissect the spectrum of molecular changes in protease and viral properties associated with resistance to TL-3, a panel of chronological in vitro escape variants was generated. We have virologically and biochemically characterized mutants with one (V82A), three (M46I/F53L/V82A), or six (L24I/M46I/F53L/L63P/V77I/V82A) changes in the protease and structurally modeled the protease mutant containing six changes. Virus containing six changes was found to be 17-fold more resistant to TL-3 in cell culture than was wild-type virus but maintained similar in vitro replication kinetics compared to the wild-type virus. Analyses of enzyme activity of protease variants with one, three, and six changes indicated that these enzymes, compared to wild-type protease, retained 40, 47, and 61% activity, respectively. These results suggest that deficient protease enzymatic activity is sufficient for function, and the observed protease restoration might imply a selective advantage, at least in vitro, for increased protease activity. PMID:11533212

Implementation of specific strength training among industrial laboratory technicians: long-term effects on back, neck and upper extremity pain.

PubMed

Pedersen, Mogens Theisen; Andersen, Christoffer H; Zebis, Mette K; Sjøgaard, Gisela; Andersen, Lars L

2013-10-09

Previous studies have shown positive effects of physical exercise at the workplace on musculoskeletal disorders. However, long-term adherence remains a challenge. The present study evaluates long-term adherence and effects of a workplace strength training intervention on back, neck and upper extremity pain among laboratory technicians. Cluster-randomized controlled trial involving 537 industrial laboratory technicians. Subjects were randomized at the cluster level to one of two groups: training group 1 (TG1, n = 282) performing supervised strength training from February to June 2009 (round one) or training group 2 (TG2, n = 255) performing supervised strength training from August to December 2009 (round two). The outcome measures were changes in self-reported pain intensity (0-9) in the back, neck and upper extremity as well as Disability of the Arm, Shoulder and Hand (DASH, 0-100). Regular adherence, defined as at least one training session per week, was achieved by around 85% in both groups in the supervised training periods. In the intention-to-treat analyses there were significant group by time effects for pain in the neck, right shoulder, right hand and lower back and DASH-resulting in significant reductions in pain (mean 0.3 to 0.5) and DASH (mean 3.9) in the scheduled training group compared to the reference group. For TG1 there were no significant changes in pain in round two, i.e. they maintained the pain reduction achieved in round one. Subgroup analyses among those with severe pain (> = 3 on a scale of 0-9) showed a significant group by time effect for pain in the neck, right shoulder, upper back and lower back. For these subgroups the pain reduction in response to training ranged from 1.1 to 1.8. Specific strength training at the workplace can lead to significant long-term reductions in spinal and upper extremity pain and DASH. The pain reductions achieved during the intensive training phase with supervision appears to be maintained a half year later.

A Dietary Intervention in Urban African Americans

PubMed Central

Miller, Edgar R.; Cooper, Lisa A.; Carson, Kathryn A.; Wang, Nae-Yuh; Appel, Lawrence J.; Gayles, Debra; Charleston, Jeanne; White, Karen; You, Na; Weng, Yingjie; Martin-Daniels, L. Michelle; Bates-Hopkins, Barbara; Robb, Inez; Franz, Whitney K.; Brown, Emily L.; Halbert, Jennifer P.; Albert, Michael C.; Dalcin, Arlene T.; Yeh, Hsin-Chieh

2015-01-01

Introduction Unhealthy diets, often low in potassium, likely contribute to racial disparities in blood pressure. We tested the effectiveness of providing weekly dietary advice, assistance with selection of higher potassium grocery items, and a $30 per week food allowance on blood pressure and other outcomes in African American adults with hypertension. Design We conducted an 8-week RCT with two parallel arms between May 2012 and November 2013. Setting/participants We randomized 123 African Americans with controlled hypertension from an urban primary care clinic in Baltimore, Maryland and implemented the trial in partnership with a community supermarket and the Baltimore City Health Department. Mean (SD) age was 58.6 (9.5) years, 71% were female, blood pressure was 131.3 (14.7)/77.2 (10.5) mmHg, BMI was 34.5 (8.2) kg/m2, and 28% had diabetes. Intervention Participants randomized to the active intervention group (Dietary Approaches to Stop Hypertension [DASH]-Plus) received coach-directed dietary advice and assistance with weekly online ordering and purchasing of high-potassium foods ($30/week) delivered by a community supermarket to a neighborhood library. Participants in the control group received a printed DASH diet brochure along with debit account of equivalent value to that of the DASH-Plus group. Main outcome measures The primary outcome was blood pressure change. Analyses were conducted in January to October 2014. Results Compared with the control group, the DASH-Plus group increased self-reported consumption of fruits and vegetables (mean=1.4, 95% CI=0.7, 2.1 servings/day), estimated intake of potassium (mean=0.4, 95% CI=0.1, 0.7 grams/day), and urine potassium excretion (mean=19%, 95% CI=1%, 38%). There was no significant effect on blood pressure. Conclusions A program providing dietary advice, assistance with grocery ordering, and $30/week of high-potassium foods in African American patients with controlled hypertension in a community-based clinic did not reduce BP. However, the intervention increased consumption of fruits, vegetables, and urinary excretion of potassium. PMID:26321012

Return to football and long-term clinical outcomes after thumb ulnar collateral ligament suture anchor repair in collegiate athletes.

PubMed

Werner, Brian C; Hadeed, Michael M; Lyons, Matthew L; Gluck, Joshua S; Diduch, David R; Chhabra, A Bobby

2014-10-01

To evaluate return to play after complete thumb ulnar collateral ligament (UCL) injury treated with suture anchor repair for both skill position and non-skill position collegiate football athletes and report minimum 2-year clinical outcomes in this population. For this retrospective study, inclusion criteria were complete rupture of the thumb UCL and suture anchor repair in a collegiate football athlete performed by a single surgeon who used an identical technique for all patients. Data collection included chart review, determination of return to play, and Quick Disabilities of the Arm, Shoulder, and Hand (QuickDASH) outcomes. A total of 18 collegiate football athletes were identified, all of whom were evaluated for follow-up by telephone, e-mail, or regular mail at an average 6-year follow-up. Nine were skill position players; the remaining 9 played in nonskill positions. All players returned to at least the same level of play. The average QuickDASH score for the entire cohort was 1 out of 100; QuickDASH work score, 0 out of 100; and sport score, 1 out of 100. Average time to surgery for skill position players was 12 days compared with 43 for non-skill position players. Average return to play for skill position players was 7 weeks postoperatively compared with 4 weeks for non-skill position players. There was no difference in average QuickDASH overall scores or subgroup scores between cohorts. Collegiate football athletes treated for thumb UCL injuries with suture anchor repair had quick return to play, reliable return to the same level of activity, and excellent long-term clinical outcomes. Skill position players had surgery sooner after injury and returned to play later than non-skill position players, with no differences in final level of play or clinical outcomes. Management of thumb UCL injuries in collegiate football athletes can be safely and effectively tailored according to the demands of the player's football position. Therapeutic IV. Copyright © 2014 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.

A Dietary Intervention in Urban African Americans: Results of the "Five Plus Nuts and Beans" Randomized Trial.

PubMed

Miller, Edgar R; Cooper, Lisa A; Carson, Kathryn A; Wang, Nae-Yuh; Appel, Lawrence J; Gayles, Debra; Charleston, Jeanne; White, Karen; You, Na; Weng, Yingjie; Martin-Daniels, Michelle; Bates-Hopkins, Barbara; Robb, Inez; Franz, Whitney K; Brown, Emily L; Halbert, Jennifer P; Albert, Michael C; Dalcin, Arlene T; Yeh, Hsin-Chieh

2016-01-01

Unhealthy diets, often low in potassium, likely contribute to racial disparities in blood pressure. We tested the effectiveness of providing weekly dietary advice, assistance with selection of higher potassium grocery items, and a $30 per week food allowance on blood pressure and other outcomes in African American adults with hypertension. We conducted an 8-week RCT with two parallel arms between May 2012 and November 2013. We randomized 123 African Americans with controlled hypertension from an urban primary care clinic in Baltimore, Maryland, and implemented the trial in partnership with a community supermarket and the Baltimore City Health Department. Mean (SD) age was 58.6 (9.5) years; 71% were female; blood pressure was 131.3 (14.7)/77.2 (10.5) mmHg; BMI was 34.5 (8.2); and 28% had diabetes. Participants randomized to the active intervention group (Dietary Approaches to Stop Hypertension [DASH]-Plus) received coach-directed dietary advice and assistance with weekly online ordering and purchasing of high-potassium foods ($30/week) delivered by a community supermarket to a neighborhood library. Participants in the control group received a printed DASH diet brochure along with a debit account of equivalent value to that of the DASH-Plus group. The primary outcome was blood pressure change. Analyses were conducted in January to October 2014. Compared with the control group, the DASH-Plus group increased self-reported consumption of fruits and vegetables (mean=1.4, 95% CI=0.7, 2.1 servings/day); estimated intake of potassium (mean=0.4, 95% CI=0.1, 0.7 grams/day); and urine potassium excretion (mean=19%, 95% CI=1%, 38%). There was no significant effect on blood pressure. A program providing dietary advice, assistance with grocery ordering, and $30/week of high-potassium foods in African American patients with controlled hypertension in a community-based clinic did not reduce BP. However, the intervention increased consumption of fruits, vegetables, and urinary excretion of potassium. Copyright © 2016 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights reserved.

The effect of a dietary portfolio compared to a DASH-type diet on blood pressure.

PubMed

Jenkins, D J A; Jones, P J; Frohlich, J; Lamarche, B; Ireland, C; Nishi, S K; Srichaikul, K; Galange, P; Pellini, C; Faulkner, D; de Souza, R J; Sievenpiper, J L; Mirrahimi, A; Jayalath, V H; Augustin, L S; Bashyam, B; Leiter, L A; Josse, R; Couture, P; Ramprasath, V; Kendall, C W C

2015-12-01

Compared to a DASH-type diet, an intensively applied dietary portfolio reduced diastolic blood pressure at 24 weeks as a secondary outcome in a previous study. Due to the importance of strategies to reduce blood pressure, we performed an exploratory analysis pooling data from intensively and routinely applied portfolio treatments from the same study to assess the effect over time on systolic, diastolic and mean arterial pressure (MAP), and the relation to sodium (Na(+)), potassium (K(+)), and portfolio components. 241 participants with hyperlipidemia, from four academic centers across Canada were randomized and completed either a DASH-type diet (control n = 82) or a dietary portfolio that included, soy protein, viscous fibers and nuts (n = 159) for 24 weeks. Fasting measures and 7-day food records were obtained at weeks 0, 12 and 24, with 24-h urines at weeks 0 and 24. The dietary portfolio reduced systolic, diastolic and mean arterial blood pressure compared to the control by 2.1 mm Hg (95% CI, 4.2 to -0.1 mm Hg) (p = 0.056), 1.8 mm Hg (CI, 3.2 to 0.4 mm Hg) (p = 0.013) and 1.9 mm Hg (CI, 3.4 to 0.4 mm Hg) (p = 0.015), respectively. Blood pressure reductions were small at 12 weeks and only reached significance at 24 weeks. Nuts, soy and viscous fiber all related negatively to change in mean arterial pressure (ρ = -0.15 to -0.17, p ≤ 0.016) as did urinary potassium (ρ = -0.25, p = 0.001), while the Na(+)/K(+) ratio was positively associated (ρ = 0.20, p = 0.010). Consumption of a cholesterol-lowering dietary portfolio also decreased blood pressure by comparison with a healthy DASH-type diet. CLINICAL TRIAL REG. NO.: NCT00438425, clinicaltrials.gov. Copyright © 2015 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.

Effects of dietary sodium on metabolites: the Dietary Approaches to Stop Hypertension (DASH)-Sodium Feeding Study.

PubMed

Derkach, Andriy; Sampson, Joshua; Joseph, Justin; Playdon, Mary C; Stolzenberg-Solomon, Rachael Z

2017-10-01

Background: High sodium intake is known to increase blood pressure and is difficult to measure in epidemiologic studies. Objective: We examined the effect of sodium intake on metabolites within the DASH (Dietary Approaches to Stop Hypertension Trial)-Sodium Trial to further our understanding of the biological effects of sodium intake beyond blood pressure. Design: The DASH-Sodium Trial randomly assigned individuals to either the DASH diet (low in fat and high in protein, low-fat dairy, and fruits and vegetables) or a control diet for 12 wk. Participants within each diet arm received, in random order, diets containing high (150 nmol or 3450 mg), medium (100 nmol or 2300 mg), and low (50 nmol or 1150 mg) amounts of sodium for 30 d (crossover design). Fasting blood samples were collected at the end of each sodium intervention. We measured 531 identified plasma metabolites in 73 participants at the end of their high- and low-sodium interventions and in 46 participants at the end of their high- and medium-sodium interventions ( N = 119). We used linear mixed-effects regression to model the relation between each log-transformed metabolite and sodium intake. We also combined the resulting P values with Fisher's method to estimate the association between sodium intake and 38 metabolic pathways or groups. Results: Six pathways were associated with sodium intake at a Bonferroni-corrected threshold of 0.0013 (e.g., fatty acid, food component or plant, benzoate, γ-glutamyl amino acid, methionine, and tryptophan). Although 82 metabolites were associated with sodium intake at a false discovery rate ≤0.10, only 4-ethylphenylsufate, a xenobiotic related to benzoate metabolism, was significant at a Bonferroni-corrected threshold ( P < 10 -5 ). Adjustment for coinciding change in blood pressure did not substantively alter the association for the top-ranked metabolites. Conclusion: Sodium intake is associated with changes in circulating metabolites, including gut microbial, tryptophan, plant component, and γ-glutamyl amino acid-related metabolites. This trial was registered at clinicaltrials.gov as NCT00000608. © 2017 American Society for Nutrition.

Return-to-activity after anatomical reconstruction of acute high-grade acromioclavicular separation.

PubMed

Saier, T; Plath, J E; Beitzel, K; Minzlaff, P; Feucht, J M; Reuter, S; Martetschläger, F; Imhoff, Andreas B; Aboalata, M; Braun, S

2016-04-02

To evaluate return-to-activity (RtA) after anatomical reconstruction of acute high-grade acromioclavicular joint (ACJ) separation. A total of 42 patients with anatomical reconstruction of acute high-grade ACJ-separation (Rockwood Type V) were surveyed to determine RtA at a mean 31 months follow-up (f-u). Sports disciplines, intensity, level of competition, participation in overhead and/or contact sports, as well as activity scales (DASH-Sport-Module, Tegner Activity Scale) were evaluated. Functional outcome evaluation included Constant score and QuickDASH. All patients (42/42) participated in sporting activities at f-u. Neither participation in overhead/contact sports, nor level of activity declined significantly (n.s.). 62 % (n = 26) of patients reported subjective sports specific ACJ integrity to be at least the same as prior to the trauma. Sporting intensity (hours/week: 7.3 h to 5.4 h, p = .004) and level of competition (p = .02) were reduced. If activity changed, in 50 % other reasons but clinical symptoms/impairment were named for modified behavior. QuickDASH (mean 6, range 0-54, SD 11) and DASH-Sport-Module (mean 6, range 0-56, SD 13) revealed only minor disabilities at f-u. Over time Constant score improved significant to an excellent score (mean 94, range 86-100, SD 4; p < .001). Functional outcome was not correlated with RtA (n.s.). All patients participated in sporting activities after anatomical reconstruction of high-grade (Rockwood Type V) ACJ-separation. With a high functional outcome there was no significant change in activity level (Tegner) and participation in overhead and/or contact sports observed. There was no correlation between functional outcome and RtA. Limiting, there were alterations in time spent for sporting activities and level of competition observed. But in 50 % those were not related to ACJ symptoms/impairment. Unrelated to successful re-established integrity and function of the ACJ it should be considered that patients decided not return-to-activity but are very content with the procedure.

Return to sports after arthroscopic capsulolabral repair using knotless suture anchors for anterior shoulder instability in soccer players: minimum 5-year follow-up study.

PubMed

Alentorn-Geli, Eduard; Álvarez-Díaz, Pedro; Doblas, Jesús; Steinbacher, Gilbert; Seijas, Roberto; Ares, Oscar; Boffa, Juan José; Cuscó, Xavier; Cugat, Ramón

2016-02-01

To report the return to sports and recurrence rates in competitive soccer players after arthroscopic capsulolabral repair using knotless suture anchors at a minimum of 5 years of follow-up. All competitive soccer players with anterior glenohumeral instability treated by arthroscopic capsulolabral repair using knotless suture anchors between 2002 and 2009 were retrospectively identified through the medical records. Inclusion criteria were: no previous surgical treatment of the involved shoulder, absence of glenoid or tuberosity fractures, absence of large Hill-Sachs or glenoid bone defect, minimum follow-up of 5 years, instability during soccer practice or games, and failure of non-surgical treatment. The charts of included players were reviewed, and a phone call was performed in a cross-sectional manner to obtain information on: current soccer, return to soccer, recurrence of instability, shoulder function (Rowe score), and disability [Quick-Disability of the Arm, Shoulder, and Hand (DASH) score and Quick-DASH Sports/Performing Arts Module]. Fifty-seven young male soccer players were finally included with a median (range) follow-up of 8 (5-10) years. Forty-nine (86 %) of the soccer players were able to return to soccer and 36 of them (73 %) at the same pre-injury level. There were 6 (10.5 %) re-dislocations in the 57 players, all of them of traumatic origin produced during soccer and other unrelated activities. The main reasons to not return to soccer were: knee injuries (two players), changes in personal life (two players), and job-related (three players). None of the players quit playing soccer because of their shoulder instability injury. The median (range) Rowe score, Quick-DASH score, and Quick-DASH sports score were 80 (25-100), 2.3 (0-12.5), and 0 (0-18.8), respectively. Competitive soccer players undergoing arthroscopic capsulolabral repair with knotless suture anchors for shoulder instability without significant bone loss demonstrate excellent return to play at mid-to-long-term follow-up, with a 10.5 % chances of re-dislocating. IV.

Presenilins and γ-Secretase: Structure, Function, and Role in Alzheimer Disease

PubMed Central

De Strooper, Bart; Iwatsubo, Takeshi; Wolfe, Michael S.

2012-01-01

Presenilins were first discovered as sites of missense mutations responsible for early-onset Alzheimer disease (AD). The encoded multipass membrane proteins were subsequently found to be the catalytic components of γ-secretases, membrane-embedded aspartyl protease complexes responsible for generating the carboxyl terminus of the amyloid β-protein (Aβ) from the amyloid protein precursor (APP). The protease complex also cleaves a variety of other type I integral membrane proteins, most notably the Notch receptor, signaling from which is involved in many cell differentiation events. Although γ-secretase is a top target for developing disease-modifying AD therapeutics, interference with Notch signaling should be avoided. Compounds that alter Aβ production by γ-secretase without affecting Notch proteolysis and signaling have been identified and are currently at various stages in the drug development pipeline. PMID:22315713

DOE Office of Scientific and Technical Information (OSTI.GOV)

Venkatraman, Srikanth; Bogen, Stephane L.; Arasappan, Ashok

Hepatitis C virus (HCV) infection is the major cause of chronic liver disease, leading to cirrhosis and hepatocellular carcinoma, which affects more than 170 million people worldwide. Currently the only therapeutic regimens are subcutaneous interferon-{alpha} or polyethylene glycol (PEG)-interferon-{alpha} alone or in combination with oral ribavirin. Although combination therapy is reasonably successful with the majority of genotypes, its efficacy against the predominant genotype (genotype 1) is moderate at best, with only about 40% of the patients showing sustained virological response. Herein, the SAR leading to the discovery of 70 (SCH 503034), a novel, potent, selective, orally bioavailable NS3 protease inhibitormore » that has been advanced to clinical trials in human beings for the treatment of hepatitis C viral infections is described. X-ray structure of inhibitor 70 complexed with the NS3 protease and biological data are also discussed.« less

Characterization and isolation of an extracellular serine protease from the tomato pathogen Colletotrichum coccodes, and it's role in pathogenicity

USGS Publications Warehouse

Redman, Regina S.; Rodriguez, Rusty J.

2002-01-01

Extracellular enzymes play an important role in the pathogenicity and virulence of phytopathogenic fungi. Several isolates of Colletotrichum coccodes causal agent of anthracnose on tomato, were screened to determine the relationship between protease activity and virulence. A direct relationship was observed between extracellular protease activity and the induction of disease symptoms of fruit and mortality in plants. Isolate Cc155 exhibited the highest protease activity after five days of growth in protease induction medium and produced an extracellular serine protease (sp78) that was 78 kDa, auto-degradative, glucose repressible, and non-glycosylated. To determine the role of sp78 in pathogenicity, a UV-induced extracellular protease deficient mutant (np155) was generated from the wildtype isolate Cc155. Np155 maintained growth rates comparable to Cc155 and produced wildtype levels of extracellular cellulase but did not produce extracellular protease. Unlike Cc155, np155 caused no disease symptoms on tomato fruit and 0% mortality on tomato seedlings. These results suggest that extracellular protease activity is required for pathogenicity and virulence of C. coccodes and that the elimination of protease activity transforms a virulent pathogen to a non-pathogenic endophyte.

Characterization and isolation of an extracellular serine protease from the tomato pathogen Colletotrichum coccodes, and it's role in pathogenicity

USGS Publications Warehouse

Redman, R.S.; Rodriguez, R.J.

2002-01-01

Extracellular enzymes play an important role in the pathogenicity and virulence of phytopathogenic fungi. Several isolates of Colletotrichum coccodes, causal agent of anthracnose on tomato, were screened to determine the relationship between protease activity and virulence. A direct relationship was observed between extracellular protease activity and the induction of disease symptoms of fruit and mortality in plants. Isolate Cc155 exhibited the highest protease activity after five days of growth in protease induction medium and produced an extracellular serine protease (sp78) that was 78 kDa, auto-degradative, glucose repressible, and non-glycosylated. To determine the role of sp78 in pathogenicity, a uv-induced extracellular protease deficient mutant (np155) was generated from the wildtype isolate Cc155. Np155 maintained growth rates comparable to Cc155 and produced wildtype levels of extracellular cellulase but did not produce extracellular protease. Unlike Cc155, np155 caused no disease symptoms on tomato fruit and 0% mortality on tomato seedlings. These results suggest that extracellular protease activity is required for pathogenicity and virulence of C. coccodes, and that the elimination of protease activity transforms a virulent pathogen to a non-pathogenic endophyte.

Purification and characterization of a cysteine protease from corms of freesia, Freesia reflacta.

PubMed

Kaneda, M; Yonezawa, H; Uchikoba, T

1997-09-01

A protease (freesia protease B) has been purified to electrophoretic homogeneity from corms of freesia, Freesia reflacta by five steps of chromatography. Its M(r) was estimated to be about 26,000 by SDS-PAGE. The optimum pH of the enzyme was 6.0-7.0 at 30 degrees C using casein as a substrate. The enzyme was strongly inhibited by p-chloromercuribenzoic acid but not by phenylmethanesulphonylfluoride and EDTA. These results indicate that freesia protease B is a cysteine protease. Nine sites of oxidized insulin B-chain were cleaved by freesia protease B in 24 h of hydrolysis. The four cleavage sites among them resembled those of papain. From the digestion of five peptidyl substrates the specificity of freesia protease B was found to be approximately broad, but the preferential cleavage sites were negatively charged residues at P1 positions. Freesia protease B preferred also the large hydrophobic amino acid residues at the P2 position, in a similar manner to papain. The amino terminal sequence of freesia protease B was identical with those of papain in regard to the conservative residues of cysteine protease.

Multi-Approach Analysis for the Identification of Proteases within Birch Pollen.

PubMed

McKenna, Olivia E; Posselt, Gernot; Briza, Peter; Lackner, Peter; Schmitt, Armin O; Gadermaier, Gabriele; Wessler, Silja; Ferreira, Fatima

2017-07-04

Birch pollen allergy is highly prevalent, with up to 100 million reported cases worldwide. Proteases in such allergen sources have been suggested to contribute to primary sensitisation and exacerbation of allergic disorders. Until now the protease content of Betula verrucosa , a birch species endemic to the northern hemisphere has not been studied in detail. Hence, we aim to identify and characterise pollen and bacteria-derived proteases found within birch pollen. The pollen transcriptome was constructed via de novo transcriptome sequencing and analysis of the proteome was achieved via mass spectrometry; a cross-comparison of the two databases was then performed. A total of 42 individual proteases were identified at the proteomic level. Further clustering of proteases into their distinct catalytic classes revealed serine, cysteine, aspartic, threonine, and metallo-proteases. Further to this, protease activity of the pollen was quantified using a fluorescently-labelled casein substrate protease assay, as 0.61 ng/mg of pollen. A large number of bacterial strains were isolated from freshly collected birch pollen and zymographic gels with gelatinase and casein, enabled visualisation of proteolytic activity of the pollen and the collected bacterial strains. We report the successful discovery of pollen and bacteria-derived proteases of Betula verrucosa .

Protease-mediated drug delivery

NASA Astrophysics Data System (ADS)

Dickson, Eva F.; Goyan, Rebecca L.; Kennedy, James C.; Mackay, M.; Mendes, M. A. K.; Pottier, Roy H.

2003-12-01

Drugs used in disease treatment can cause damage to both malignant and normal tissue. This toxicity limits the maximum therapeutic dose. Drug targeting is of high interest to increase the therapeutic efficacy of the drug without increasing systemic toxicity. Certain tissue abnormalities, disease processes, cancers, and infections are characterized by high levels of activity of specific extracellular and/or intracellular proteases. Abnormally high activity levels of specific proteases are present at sites of physical or chemical trauma, blood clots, malignant tumors, rheumatoid arthritis, inflammatory bowel disease, gingival disease, glomerulonerphritis, and acute pancreatitis. Abnormal protease activity is suspected in development of liver thrombosis, pulmonary emphysema, atherosclerosis, and muscular dystrophy. Inactiviating disease-associated proteases by the administration of appropriate protease inhibitors has had limited success. Instead, one could use such proteases to target drugs to treat the condition. Protease mediated drug delivery offers such a possibility. Solubilizing groups are attached to insoluble drugs via a polypeptide chain which is specifically cleavable by certian proteases. When the solubilized drug enounters the protease, the solubilizing moieties are cleaved, and the drug precipitates at the disease location. Thus, a smaller systemic dosage could result in a therapeutic drug concentration at the treatment site with less systemic toxicity.

The herpesvirus proteases as targets for antiviral chemotherapy.

PubMed

Waxman, L; Darke, P L

2000-01-01

Viruses of the family Herpesviridae are responsible for a diverse set of human diseases. The available treatments are largely ineffective, with the exception of a few drugs for treatment of herpes simplex virus (HSV) infections. For several members of this DNA virus family, advances have been made recently in the biochemistry and structural biology of the essential viral protease, revealing common features that may be possible to exploit in the development of a new class of anti-herpesvirus agents. The herpesvirus proteases have been identified as belonging to a unique class of serine protease, with a Ser-His-His catalytic triad. A new, single domain protein fold has been determined by X-ray crystallography for the proteases of at least three different herpesviruses. Also unique for serine proteases, dimerization has been shown to be required for activity of the cytomegalovirus and HSV proteases. The dimerization requirement seriously impacts methods needed for productive, functional analysis and inhibitor discovery. The conserved functional and catalytic properties of the herpesvirus proteases lead to common considerations for this group of proteases in the early phases of inhibitor discovery. In general, classical serine protease inhibitors that react with active site residues do not readily inactivate the herpesvirus proteases. There has been progress however, with activated carbonyls that exploit the selective nucleophilicity of the active site serine. In addition, screening of chemical libraries has yielded novel structures as starting points for drug development. Recent crystal structures of the herpesvirus proteases now allow more direct interpretation of ligand structure-activity relationships. This review first describes basic functional aspects of herpesvirus protease biology and enzymology. Then we discuss inhibitors identified to date and the prospects for their future development.

Exogenous Thyropin from p41 Invariant Chain Diminishes Cysteine Protease Activity and Affects IL-12 Secretion during Maturation of Human Dendritic Cells

PubMed Central

Zavašnik-Bergant, Tina; Bergant Marušič, Martina

2016-01-01

Dendritic cells (DC) play a pivotal role as antigen presenting cells (APC) and their maturation is crucial for effectively eliciting an antigen-specific immune response. The p41 splice variant of MHC class II-associated chaperone, called invariant chain p41 Ii, contains an amino acid sequence, the p41 fragment, which is a thyropin-type inhibitor of proteolytic enzymes. The effects of exogenous p41 fragment and related thyropin inhibitors acting on human immune cells have not been reported yet. In this study we demonstrate that exogenous p41 fragment can enter the endocytic pathway of targeted human immature DC. Internalized p41 fragment has contributed to the total amount of the immunogold labelled p41 Ii-specific epitope, as quantified by transmission electron microscopy, in particular in late endocytic compartments with multivesicular morphology where antigen processing and binding to MHC II take place. In cell lysates of treated immature DC, diminished enzymatic activity of cysteine proteases has been confirmed. Internalized exogenous p41 fragment did not affect the perinuclear clustering of acidic cathepsin S-positive vesicles typical of mature DC. p41 fragment is shown to interfere with the nuclear translocation of NF-κB p65 subunit in LPS-stimulated DC. p41 fragment is also shown to reduce the secretion of interleukin-12 (IL-12/p70) during the subsequent maturation of treated DC. The inhibition of proteolytic activity of lysosomal cysteine proteases in immature DC and the diminished capability of DC to produce IL-12 upon their subsequent maturation support the immunomodulatory potential of the examined thyropin from p41 Ii. PMID:26960148

N-glycans of Human Protein C Inhibitor: Tissue-Specific Expression and Function

PubMed Central

Engström, Åke; Sooriyaarachchi, Sanjeewani; Ubhayasekera, Wimal; Hreinsson, Julius; Wånggren, Kjell; Clark, Gary F.; Dell, Anne; Schedin-Weiss, Sophia

2011-01-01

Protein C inhibitor (PCI) is a serpin type of serine protease inhibitor that is found in many tissues and fluids in human, including blood plasma, seminal plasma and urine. This inhibitor displays an unusually broad protease specificity compared with other serpins. Previous studies have shown that the N-glycan(s) and the NH2-terminus affect some blood-related functions of PCI. In this study, we have for the first time determined the N-glycan profile of seminal plasma PCI, by mass spectrometry. The N-glycan structures differed markedly compared with those of both blood-derived and urinary PCI, providing evidence that the N-glycans of PCI are expressed in a tissue-specific manner. The most abundant structure (m/z 2592.9) had a composition of Fuc3Hex5HexNAc4, consistent with a core fucosylated bi-antennary glycan with terminal Lewisx. A major serine protease in semen, prostate specific antigen (PSA), was used to evaluate the effects of N-glycans and the NH2-terminus on a PCI function related to the reproductive tract. Second-order rate constants for PSA inhibition by PCI were 4.3±0.2 and 4.1±0.5 M−1s−1 for the natural full-length PCI and a form lacking six amino acids at the NH2-terminus, respectively, whereas these constants were 4.8±0.1 and 29±7 M−1s−1 for the corresponding PNGase F-treated forms. The 7–8-fold higher rate constants obtained when both the N-glycans and the NH2-terminus had been removed suggest that these structures jointly affect the rate of PSA inhibition, presumably by together hindering conformational changes of PCI required to bind to the catalytic pocket of PSA. PMID:22205989

Discovery of potent peptide-mimetic antagonists for the human thrombin receptor, protease-activated receptor-1 (PAR-1).

PubMed

Maryanoff, Bruce E; Zhang, Han-Cheng; Andrade-Gordon, Patricia; Derian, Claudia K

2003-03-01

Protease-activated receptors (PARs) represent a unique family of seven-transmembrane G-protein-coupled receptors, which are enzymatically cleaved to expose a new extracellular N-terminus that acts as a tethered activating ligand. PAR-1 is cleaved and activated by the serine protease alpha-thrombin, is expressed in various tissues (e.g. platelets and vascular cells), and is involved in cellular responses associated with hemostasis, proliferation, and tissue injury. By using a de novo design approach, we have discovered a series of potent heterocycle-based peptide-miimetic antagonists of PAR-1, exemplified by advanced leads RWJ-56110 (22) and RWJ-58259 (32). These compounds are potent, selective PAR-1 antagonists, devoid of PAR-1 agonist and thrombin inhibitory activity: they bind to PAR-1, interfere with calcium mobilization and cellular functions associated with PAR-1, and do not affect PAR-2, PAR-3, or PAR-4. RWJ-56110 was determined to be a direct inhibitor of PAR-1 activation and internalization, without affecting PAR-1 N-terminal cleavage. At high concentrations of alpha-thrombin, RWJ-56110 fully blocked activation responses in human vascular cells, but not in human platelets; whereas, at high concentrations of TRAP-6, RWJ-56110 blocked activation responses in both cell types. This result is consistent with the presence of another thrombin receptor on human platelets, namely PAR-4. RWJ-56110 and RWJ-58259 clearly interrupt the binding of a tethered ligand to its receptor. RWJ-58259 demonstrated antirestenotic activity in a rat balloon angioplasty model and antithrombotic activity in a cynomolgus monkey arterial injury model. Such PAR-1 antagonists should not only serve as useful tools to delineate the physiological and pathophysiological roles of PAR-1, but also may have therapeutic potential for treating thrombosis and restenosis in humans.

Studies on detection and analysis of proteases in leaf extract of medicinally important plants.

PubMed

Chinnadurai, Gandhi Shree; Krishnan, Sivakumar; Perumal, Palani

2018-02-01

The whole plant or the extracts obtained from them have long been used as medicine to treat various human diseases and disorders. Notably, those plants endowed with protease activity have been traditionally used as the agents for treating tumors, digestion disorders, swelling, blood coagulation, fibrinolysis and also for immune-modulation. Proteases occupy a pivotal position in enzyme based industries. Plant proteases have been increasingly exploited for pharmaceutical, food, leather and textile processing industries. Earlier investigations have focused on the occurrence of proteases in medicinally unimportant plants. Therefore it has been aimed to study the occurrence of proteolytic enzymes from medicinally important plants establish any correlation exists between protease activity and medicinal use of individual plants. Crude extract were obtained from the leaves of 80 different medicinal plants. Tris-HCl buffer was used as the extraction buffer and the supernatants obtained were used for determination of total protein and protease activity using spectrophotometric methods. Qualitative screening for the presence of protease was carried out with agar diffusion method by incorporating the substrate. SDS-PAGE was used to analyse the isoforms of protease and for determination of relative molecular mass. Relatively higher protease activities were observed in the extracts of leaves of Pongamia pinnata (Fabaceae), Wrightia tinctoria (Apocyanaceae) Acalypha indica (Euphorbiaceae), Adhatoda vasica (Acanthaceae) and Curcuma longa (Zingiberaceae). No correlation was found between the total protein content and protease activity in individual plant species. SDS-PAGE analysis indicated the presence of multiple forms of protease of higher molecular weight range in several plant species. We found a strong correlation between the protease activity and medicinal application of the plant CONCLUSION: The present study has unequivocally revealed that the leaves of medicinal plants could serve as excellent sources of proteases which could be exploited for various industrial, food and pharmaceutical applications. Copyright © 2018 Elsevier GmbH. All rights reserved.