Wu, Gongqing; Yi, Yunhong
The bacterium Photorhabdus luminescens produces a number of insecticidal proteins to kill its larval prey. In this study, we cloned the gene coding for a binary toxin PirA1B1 and purified the recombinant protein using affinity chromatography combined with desalination technology. Furthermore, the cytotoxicity of the recombinant protein against the haemocytes of Galleria mellonella larvae was investigated. We found that the protein had haemocoel insecticidal activity against G. mellonella with an LD50 of 131.5 ng/larva. Intrahaemocoelic injection of PirA1B1 into G. mellonella resulted in significant decreases in haemocyte number and phagocytic ability. In in vitro experiments, PirA1B1 inhibited the spreading behaviour of the haemocytes of G. mellonella larvae and even caused haemocyte degeneration. Fluorescence microscope analysis and visualization of haemocyte F-actin stained with phalloidin-FITC showed that the PirA1B1 toxin disrupted the organization of the haemocyte cytoskeleton. Our results demonstrated that the PirA1B1 toxin disarmed the insect cellular immune system.
Wu, Gongqing; Yi, Yunhong
The bacterium Photorhabdus luminescens produces a number of insecticidal proteins to kill its larval prey. In this study, we cloned the gene coding for a binary toxin PirA1B1 and purified the recombinant protein using affinity chromatography combined with desalination technology. Furthermore, the cytotoxicity of the recombinant protein against the haemocytes of Galleria mellonella larvae was investigated. We found that the protein had haemocoel insecticidal activity against G. mellonella with an LD50 of 131.5 ng/larva. Intrahaemocoelic injection of PirA1B1 into G. mellonella resulted in significant decreases in haemocyte number and phagocytic ability. In in vitro experiments, PirA1B1 inhibited the spreading behaviour of the haemocytes of G. mellonella larvae and even caused haemocyte degeneration. Fluorescence microscope analysis and visualization of haemocyte F-actin stained with phalloidin-FITC showed that the PirA1B1 toxin disrupted the organization of the haemocyte cytoskeleton. Our results demonstrated that the PirA1B1 toxin disarmed the insect cellular immune system. PMID:27734915
Wu, Gongqing; Yi, Yunhong; Sun, Jianyu; Li, Mei; Qiu, Lihong
There is accumulating evidence that many invertebrates including insects can acquire enhanced immune protection against subsequently pathogens infection through immune priming. However, whether the toxin protein from pathogenic bacteria can induce such priming response remains unclear. Here we cloned, expressed and purified the toxin Photorhabdus insect-related proteins A2B2 (PirA2B2) from Photorhabdus luminescens TT01. We primed Galleria mellonella with sublethal dose of PirA2B2 and then challenged the larvae with viable P. luminescens TT01 at 48 h after priming. We found no evidence for immune priming in G. mellonella larvae exposed to PirA2B2. Priming the larvae with PirA2B2 did not improve their resistance in a subsequent challenge with P. luminescens TT01. Whereas a robust priming response was observed when the larvae exposed to lipopolysaccharide (LPS) extracted from P. luminescens TT01. Because the larvae primed with LPS showed significant higher resistance against P. luminescens TT01 infection than those of the PBS and BSA controls. Furthermore, we investigated the changes of the cellular immune parameters, such as hemocyte counts, phagocytic activity and encapsulation ability of the hemocytes, after priming. We found that the toxin PirA2B2 significantly decreased the cellular immunity of the larvae, whereas the LPS significantly increased them. These results indicated that the degree of priming response in G. mellonella correlated positively to the levels of cellular immune parameters, and the underlying mechanism in regulating the immune priming of invertebrates was not homologous to that of the immunological memory of vertebrates.
Singh, O P; Dykes, C L; Sharma, G; Das, M K
Leucine-to-phenylalanine substitution at residue L1014 in the voltage-gated sodium channel, target site of action for dichlorodiphenyltrichloroethane (DDT) and pyrethroids, is the most common knockdown resistance (kdr) mutation reported in several insects conferring resistance against DDT and pyrethroids. Here, we report presence of two coexisting alternative transversions, A>T and A>C, on the third codon position of L1014 residue in malaria vector Anopheles subpictus Grassi (species A) from Jamshedpur (India), both leading to the same amino acid substitution of Leu-to-Phe with allelic frequencies of 19 and 67%, respectively. A single primer-introduced restriction analysis-polymerase chain reaction (PIRA-PCR) was devised for the identification of L1014F-kdr mutation in An. subpictus resulting from either type of point mutation. Genotyping of samples with PIRA-PCR revealed high frequency (82%) of L1014F-kdr mutation in the study area.
Background Mexico has the highest adult overweight and obesity prevalence in the Americas; 23.8% of children <5 years old are at risk for overweight and 9.7% are already overweight or obese. Creciendo Sanos was a pilot intervention to prevent obesity among preschoolers in Instituto Mexicano del Seguro Social (IMSS) clinics. Methods We randomized 4 IMSS primary care clinics to either 6 weekly educational sessions promoting healthful nutrition and physical activity or usual care. We recruited 306 parent-child pairs: 168 intervention, 138 usual care. Children were 2-5 years old with WHO body mass index (BMI) z-score 0-3. We measured children’s height and weight and parents reported children’s diet and physical activity at baseline and 3 and 6-month follow-up. We analyzed behavioral and BMI outcomes with generalized mixed models incorporating multiple imputation for missing values. Results 93 (55%) intervention and 96 (70%) usual care families completed 3 and 6-month follow-up. At 3 months, intervention v. usual care children increased vegetables by 6.3 servings/week (95% CI, 1.8, 10.8). In stratified analyses, intervention participants with high program adherence (5-6 sessions) decreased snacks and screen time and increased vegetables v. usual care. No further effects on behavioral outcomes or BMI were observed. Transportation time and expenses were barriers to adherence. 90% of parents who completed the post-intervention survey were satisfied with the program. Conclusions Although satisfaction was high among participants, barriers to participation and retention included transportation cost and time. In intention to treat analyses, we found intervention effects on vegetable intake, but not other behaviors or BMI. Trial registration ClinicalTrials.gov NCT01539070. Comisión Nacional de Investigación Científica del IMSS: 2009-785-120. PMID:24649831
Using a lighthearted and simple approach, this 23-minute videotape in English and Spanish versions presents interactions between parents and children while reading books. The children in the videotape range in age from 0 to 5 years. The video is introduced by scenes of children enjoying books while narration discussing the impact of reading to…
Office of Elementary and Secondary Education (ED), Washington, DC. Safe and Drug Free Schools Program.
This publication is part of the Department of Education's ongoing effort to provide information to emerging populations in their native language on how to create and maintain drug-free and safe schools across the nation. Throughout the publication are personal stories and artwork of elementary and secondary school students that promote the message…
The bibliography contains citations concerning chlorofluorocarbons (CFC's), and efforts to find substitute materials in response to the need to lessen the environmental impact of CFC on the ozone layer, as agreed to in the international Montreal Protocol. CFC alternatives are expected to be used as aerosol propellants, blowing agents, refrigerants, fire extinguishants, and solvents. References to the effects of CFC on ozone are found in separate bibliographies. (Contains 250 citations and includes a subject term index and title list.)
El-Houssini, Ola M; Zawilla, Nagwan H; Mohammad, Mohammad A
Specific stability indicating reverse-phase liquid chromatography (RP-LC) assay method (SIAM) was developed for the determination of cinnarizine (Cinn)/piracetam (Pira) and cinnarizine (Cinn)/heptaminol acefyllinate (Hept) in the presence of the reported degradation products of Cinn. A C18 column and gradient mobile phase was applied for good resolution of all peaks. The detection was achieved at 210 nm and 254 nm for Cinn/Pira and Cinn/Hept, respectively. The responses were linear over concentration ranges of 20-200, 20-1000 and 25-1000 μgmL(-1) for Cinn, Pira, and Hept respectively. The proposed method was validated for linearity, accuracy, repeatability, intermediate precision, and robustness via statistical analysis of the data. The method was shown to be precise, accurate, reproducible, sensitive, and selective for the analysis of Cinn/Pira and Cinn/Hept in laboratory prepared mixtures and in pharmaceutical formulations.
EL-Houssini, Ola M.; Zawilla, Nagwan H.; Mohammad, Mohammad A.
Specific stability indicating reverse-phase liquid chromatography (RP-LC) assay method (SIAM) was developed for the determination of cinnarizine (Cinn)/piracetam (Pira) and cinnarizine (Cinn)/heptaminol acefyllinate (Hept) in the presence of the reported degradation products of Cinn. A C18 column and gradient mobile phase was applied for good resolution of all peaks. The detection was achieved at 210 nm and 254 nm for Cinn/Pira and Cinn/Hept, respectively. The responses were linear over concentration ranges of 20–200, 20–1000 and 25–1000 μgmL−1 for Cinn, Pira, and Hept respectively. The proposed method was validated for linearity, accuracy, repeatability, intermediate precision, and robustness via statistical analysis of the data. The method was shown to be precise, accurate, reproducible, sensitive, and selective for the analysis of Cinn/Pira and Cinn/Hept in laboratory prepared mixtures and in pharmaceutical formulations. PMID:24137049
Jo, Yong Hun; Patnaik, Bharat Bhusan; Kang, Se Won; Chae, Sung-Hwa; Oh, Seunghan; Kim, Dong Hyun; Noh, Mi Young; Seo, Gi Won; Jeong, Heon Cheon; Noh, Ju Young; Jeong, Ji Eun; Hwang, Hee Ju; Ko, Kisung; Han, Yeon Soo; Lee, Yong Seok
Background Most traditional genome sequencing projects involving viruses include the culture and purification of the virus particles. However, purification of virions may yield insufficient material for traditional sequencing. The electrophoretic method described here provides a strategy whereby the genomic DNA of the Korean isolate of Pieris rapae granulovirus (PiraGV-K) could be recovered in sufficient amounts for sequencing by purifying it directly from total host DNA by pulse-field gel electrophoresis (PFGE). Methodology/Principal Findings The total genomic DNA of infected P. rapae was embedded in agarose plugs, treated with restriction nuclease and methylase, and then PFGE was used to separate PiraGV-K DNA from the DNA of P. rapae, followed by mapping of fosmid clones of the purified viral DNA. The double-stranded circular genome of PiraGV-K was found to encode 120 open reading frames (ORFs), which covered 92% of the sequence. BLAST and ORF arrangement showed the presence of 78 homologs to other genes in the database. The mean overall amino acid identity of PiraGV-K ORFs was highest with the Chinese isolate of PiraGV (∼99%), followed up with Choristoneura occidentalis ORFs at 58%. PiraGV-K ORFs were grouped, according to function, into 10 genes involved in transcription, 11 involved in replication, 25 structural protein genes, and 15 auxiliary genes. Genes for Chitinase (ORF 10) and cathepsin (ORF 11), involved in the liquefaction of the host, were found in the genome. Conclusions/Significance The recovery of PiraGV-K DNA genome by pulse-field electrophoretic separation from host genomic DNA had several advantages, compared with its isolation from particles harvested as virions or inclusions from the P. rapae host. We have sequenced and analyzed the 108,658 bp PiraGV-K genome purified by the electrophoretic method. The method appears to be generally applicable to the analysis of genomes of large viruses. PMID:24391907
Tristán-González, Margarito; Labarthe-Hernández, Guillermo; Aguirre-Díaz, Gerardo J.; Aguillón-Robles, Alfredo
The volcano-tectonic events at the Villa de Reyes Graben (VRG), in the southern Sierra Madre Occidental, Mexico, include 1) a regional NNE fault system developed before 32 Ma, 2) this pre-32 Ma faulting controlled the emplacement of 31.5 Ma dacitic domes, 3) NE faulting at 28 Ma that displaced the 31.5 Ma dacitic domes and formed the VRG, as well as the oblique grabens of Bledos and Enramadas oriented NW, 4) emplacement of Panalillo ignimbrite at 28 Ma filling the VRG and erupting from fissures related to the oblique grabens, and eruption of Placa basalt apparently also from fault-controlled vents.
Organization NIF National Islamic Front PIJ Palestinian Islamic Jihad PIRA Provisional Irish Republican Army WMD Weapons of Mass Destruction xii THIS PAGE...Islamic Front ( NIF ) to power. The party’s leader, Hassan al-Turabi, sought to establish an Islamist state in Sudan and offered sanctuary for Islamic
Liu, Saijun; Chen, Liyuan; Zhang, Xiandong; Li, Jian; Lin, Haiying; Liu, Louhui; Xie, Jiansheng; Ge, Huijuan; Ye, Minglan; Chen, Caifen; Ji, Xingwen; Zhang, Caifen; Xu, Fengping; Jiang, Hui; Zhen, Hefu; Chen, Shiping; Wang, Wei
We have developed a new method for non-invasive prenatal testing (NIPT) of paternally inherited fetal mutants for β-thalassemia (β-thal). Specially designed primer-introduced restriction analysis-polymerase chain reaction (PIRA-PCR) were used to detect four major mutations [IVS-II-654, HBB: c.316-197C > T; codon 17 (A > T), HBB: c.52A > T; -28 (A > G), HBB: c.-78A > G and codons 41/42 (-TTCT), HBB: c.126_129delCTTT] causing β-thal in China. The PIRA-PCR assay was first tested in a series of mixed DNA with different concentrations and mixed proportions. Subsequently, this assay was further tested in 10 plasma DNA samples collected from pregnant women. In the DNA mixture simulation test, the PIRA-PCR assay was able to detect 3.0% target genomic DNA (gDNA) mixed in 97.0% wild-type gDNA isolated from whole blood. For plasma DNA testing, the results detected by PIRA-PCR assay achieved 100.0% consistency with those obtained from the amniocentesis analysis. This new method could potentially be used for NIPT of paternally inherited fetal mutants for β-thal.
Alley, T L; Cooper, M D; Chen, M; Kubagawa, H
The genes encoding the murine paired immunoglobulin-like receptors PIR-A and PIR-B are members of a novel gene family which encode cell-surface receptors bearing immunoreceptor tyrosine-based inhibitory motifs (ITIMs) and their non-inhibitory/activatory counterparts. PIR-A and PIR-B have highly homologous extracellular domains but distinct transmembrane and cytoplasmic regions. A charged arginine in the transmembrane region of PIR-A suggests its potential association with other transmembrane proteins to form a signal transducing unit. PIR-B, in contrast, has an uncharged transmembrane region and several ITIMs in its cytoplasmic tail. These characteristics suggest that PIR-A and PIR-B which are coordinately expressed by B cells and myeloid cells, serve counter-regulatory roles in humoral and inflammatory responses. In the present study we have determined the genomic structure of the single copy PIR-B gene. The gene consists of 15 exons and spans approximately 8 kilobases. The first exon contains the 5' untranslated region, the ATG translation start site, and approximately half of the leader peptide sequence. The remainder of the leader peptide sequence is encoded by exon 2. Exons 3-8 encode the six extracellular immunoglobulin-like domains and exons 9 and 10 code for the extracellular membrane proximal and transmembrane regions. The final five exons (exons 11-15) encode for the ITIM-bearing cytoplasmic tail and the 3' untranslated region. The intron/exon boundaries of PIR-B obey the GT-AG rule and are in phase I, with the notable exception of the three boundaries determined for ITIM-containing exons. A microsatellite composed of the trinucleotide repeat AAG in the intron between exons 9 and 10 provides a useful marker for studying population genetics.
Gianfranco Piras May 2009 C on st ru ct io n E n gi n ee ri n g R es ea rc h L ab or at or y Approved for public release; distribution is... GIS operations. Julia Koschinsky provided logistical support. The report and its findings are those of the authors alone, and should not be attributed...news sources. While precise geocoding was not possible, there was sufficient information in the reports to associate each event with a neighborhood. A
Asal, Victor; Gill, Paul; Rethemeyer, R Karl; Horgan, John
This paper presents an analysis of the Provisional Irish Republican Army's (PIRA) brigade level behavior during the Northern Ireland Conflict (1970-1998) and identifies the organizational factors that impact a brigade's lethality as measured via terrorist attacks. Key independent variables include levels of technical expertise, cadre age, counter-terrorism policies experienced, brigade size, and IED components and delivery methods. We find that technical expertise within a brigade allows for careful IED usage, which significantly minimizes civilian casualties (a specific strategic goal of PIRA) while increasing the ability to kill more high value targets with IEDs. Lethal counter-terrorism events also significantly affect a brigade's likelihood of killing both civilians and high-value targets but in different ways. Killing PIRA members significantly decreases IED fatalities but also significantly decreases the possibility of zero civilian IED-related deaths in a given year. Killing innocent Catholics in a Brigade's county significantly increases total and civilian IED fatalities. Together the results suggest the necessity to analyze dynamic situational variables that impact terrorist group behavior at the sub-unit level.
Asal, Victor; Rethemeyer, R. Karl; Horgan, John
This paper presents an analysis of the Provisional Irish Republican Army's (PIRA) brigade level behavior during the Northern Ireland Conflict (1970-1998) and identifies the organizational factors that impact a brigade's lethality as measured via terrorist attacks. Key independent variables include levels of technical expertise, cadre age, counter-terrorism policies experienced, brigade size, and IED components and delivery methods. We find that technical expertise within a brigade allows for careful IED usage, which significantly minimizes civilian casualties (a specific strategic goal of PIRA) while increasing the ability to kill more high value targets with IEDs. Lethal counter-terrorism events also significantly affect a brigade's likelihood of killing both civilians and high-value targets but in different ways. Killing PIRA members significantly decreases IED fatalities but also significantly decreases the possibility of zero civilian IED-related deaths in a given year. Killing innocent Catholics in a Brigade's county significantly increases total and civilian IED fatalities. Together the results suggest the necessity to analyze dynamic situational variables that impact terrorist group behavior at the sub-unit level. PMID:25838603
Adhya, Sriya; Noé, John W.
The `Mirage' (Opti-Gone International) is a well-known optics demonstration (PIRA index number 6A20.35) that uses two opposed concave mirrors to project a real image of a small object into space. We studied image formation in the Mirage by standard 2x2 matrix methods and by exact ray tracing, with particular attention to additional real images that can be observed when the mirror separation is increased beyond one focal length. We find that the three readily observed secondary images correspond to 4, 6, or 8 reflections, respectively, contrary to previous reports.
Rahman, Mohammad M; Shoubudani, Tomoaki; Mizukami, Keijiro; Chang, Hye-Sook; Hossain, Mohammad A; Yabuki, Akira; Mitani, Sawane; Higo, Takashi; Arai, Toshiro; Yamato, Osamu
Polymerase chain reaction (PCR)-based assays combined with microchip electrophoresis were developed and evaluated for diagnosis and genotyping of GM2 gangliosidosis variant 0 (Sandhoff-like disease) in Japanese domestic cats. A preliminary genotyping survey was carried out in the population of Japanese domestic cats (1,015 cats in total) in southern Japan. Three kinds of assays including PCR primer-induced restriction analysis (PIRA) and mutagenically separated (MS)-PCR were carried out using blood-stained Flinders Technology Associates filter papers (FTA cards) as templates. The PCR products were analyzed by both agarose gel and microchip electrophoreses. All assays were sufficient to determine the genotypes of this disease, but MS-PCR offered the most rapid and simplest test, as it does not need the restriction enzyme step required in PCR-PIRA. The use of microchip electrophoresis in combination with FTA cards for sampling could shorten the time required for genotyping and simplify the procedure as well. The genotyping survey in the current study did not find any cats that possessed the mutant allele, suggesting that the prevalence of this allele is low (<0.1%) in southern Japan.
Ferreira, João A; Bila, Danielle M; Ritter, Elisabeth; Braga, Ana Cs
The disposal of healthcare waste (HCW) seems to have been solved in developed countries, while in most developing countries the problem persists because the disposal methods are expensive and larger than the budget of small- and medium-sized municipalities. The current study evaluates the encapsulation process for the disposal of medical chemical waste. The experiment was developed in the Piraí municipality (Rio de Janeiro State, Brazil) and the chemical wastes were produced in the local public hospital, as well as the 12 units of primary care health services. Chemical waste generated at health services units may include the liquid waste from cleaning materials and disinfectants, expired and unused pharmaceutical products, and cytotoxins. These are all considered hazardous waste products and they must be disposed of via an authorised system at approved sites (e.g. industrial landfills). The process of encapsulating chemical medical waste in concrete (cement, crushed stones and sand) followed by their disposal at sanitary landfills is a procedure that is not considered in Brazilian Legislation. Despite the oversight, this method of disposal was used in the municipality of Piraí, with the approval of the Rio de Janeiro State Agency for Environmental Control. The safety aspects of this method and the limits of its applicability, particularly in small municipalities, were evaluated in this study. The results indicate that, within certain parameters, this method may provide a viable solution for the disposal of HCW in small municipalities.
Bosio, D; Coggiola, M; Baracco, A; Andreis, P; Perrelli, F
Audiogram classification is crucial for hearing protection of workers occupationally exposed to noise. The methods that have been proposed are based on two principles: the morphological evaluation of the audiometric curve (eg. Merluzzi-Pira-Bosio--MPB) or the average hearing loss on different frequencies (eg. Albera-Beatrice--AB). The purpose of this study was to classify audiograms compatible with chronic acoustic trauma performed at the Occupational Medicine Outpatient Clinic of CTO Hospital in Turin from 2004 to 2011 with the methods outlined in Guidelines published by SIMLII. A substantial agreement among the methods was observed. While MPB is the most appropriate method for secondary prevention, the AB would seem more appropriate for the verification of a permanent weakening that has to be reported to the competent legal authorities.
Enquist, Lynn W.
John N. Brady, Chief of the Virus Tumor Biology Section of the Laboratory of Cellular Oncology, National Institutes of Health, died of cancer on 27 April 2009. John was a stellar member of the virology community. He was a longtime Journal of Virology reviewer and a member of the editorial board. He will be missed. Fatah Kashanchi of the George Washington University Medical Center has written John's memorial. Fatah worked with John at the NIH and published more than 30 papers with him. Fatah thanks all the people who contributed to John's obituary, including Kuan-Teh Jeang, Lou Laimins, Mary Loeken, Renaud Mehieux, Paul Lambert, Graziella Piras, Scott Gitlin, Paul Lindholm, Nadia Rosenthal, Sergi Nekhai, Brian Wigdahl, David Price, Susan J. Marriott, Cynthia Masison, Jurgen Dittmer, Eric Verdin, Bassel E. Sawaya, and John's longtime assistants Janet Duvall Grimm and Michael Radonovich, who gave immense support to all the individuals who went through John's lab. PMID:19474098
Most new teachers walk into their new classroom territory and find at least some equipment. Perhaps there are items there that you don't recognize. One way to get those types of questions answered is to post a question and/or picture on the TAP-L (Teaching Apparatus List) listserv (see: http://www.pira-online.org for more information). Within a few short minutes, the sender will receive at least a couple of responses. But what if a new teacher walks into a totally empty prep room? Now that teacher is tasked with purchasing useful equipment, and we know that the opportunity for a similar acquisition in the near future is slim. So you better choose wisely! Whether my new stockroom is well equipped or completely empty, I would make sure to have the following six items in my arsenal.
Tejera Pérez, Cristina; Bellido Guerrero, Diego
Twitter es una red social tipo microbloggingque se basa en la generación de mensajes o tuits limitados a 140 caracteres que pueden ir acompañados de enlaces y fotografías. Desde 2006, esta red ha ido creciendo, acumulando más de 300 millones de usuarios activos, que tuitean 65 millones de mensajes al día y vehiculan más de 800.000 peticiones de búsqueda en esas 24 horas. En esta inmediatez y brevedad reside su éxito. Se trata de una gran plataforma de comunicación que conecta a emisor y receptor y los agrupa según sus intereses. Cada tema o topicse identifica con una o varias palabras clave, lo que se denomina #hashtagy posibilita seguir el ritmo de la conversación tanto en directo como en diferido.
Lazardi, K; Urbina, J A; de Souza, W
We report the ultrastructural alterations induced during the proliferative stages of Trypanosoma (Schizotrypanum) cruzi, the causative agent of Chagas' disease, by two ergosterol biosynthesis inhibitors, ketoconazole and terbinafine, which had previously been shown to be potent growth inhibitors whose effects are potentiated when used in combination (J. A. Urbina, K. Lazardi, T. Aguirre, M. M. Piras, and R. Piras, Antimicrob. Agents Chemother. 32:1237-1242, 1988). Epimastigotes treated with a low concentration of ketoconazole (1 microM), which blocks ergosterol biosynthesis at the level of C-14 demethylation of lanosterol and induces cell lysis coincident with total ergosterol depletion, showed gross alterations of the kinetoplast-mitochondrion complex, which swelled and lost the organization of its inner membrane and the electron-dense bodies of its matrix. Thus, coincident with the beginning of cell lysis, the kinetoplast-mitochondrion complex occupied greater than 80% of the cell volume, while other subcellular structures such as the nucleus and subpellicular microtubules were not affected. Terbinafine, which blocks ergosterol synthesis in these cells at the level of squalene synthetase and thus leads to almost immediate arrest of growth at concentrations greater than 1 microM, produced proliferation of glycosomelike bodies, binucleated cells (arrest at cytokinesis), and eventually massive vacuolization. When the drugs were combined, the predominant effect was mitochondrial swelling, which was more drastic and took place earlier than that observed in cells treated with ketoconazole alone. In amastigotes proliferating in Vero cells, ketoconazole at the concentration required to eradicate the parasites (10 nM) produced mitochondrial swelling, the appearance of autophagic vacuoles containing partially degraded subcellular material, and finally a general breakdown of the subcellular structures. Terbinafine at 3 microM induced more limited ultrastructural damage to
Li, Yusheng; Hu, Xiaofeng; Zhang, Xu; Liu, Zhengqiang; Ding, Xuezhi; Xia, Liqiu; Hu, Shengbiao
The binary toxin 'Photorhabdus insect-related' proteins (PirAB) produced by Photorhabdus luminescens have been reported to possess both injectable and oral activities against a range of insects. Here, PirAB-fusion protein was constructed by linking pirA and pirB genes with the flexible linker (Gly4 Ser)3 DNA encoding sequence and then efficiently expressed in Escherichia coli. To better understand the role of PirAB toxin played in the process of invasion, its cytotoxicity against insect midgut CF-203 cells was investigated. Application of purified PirAB-fusion protein as well as PirA/PirB mixture caused loss of viability of CF-203 cells after 24 h incubation. CF-203 cells treated by PirAB-fusion protein displayed morphological changes typical of apoptosis, such as cell shrinkage, cell membrane blebbing, nuclear condensation and DNA fragmentation. Moreover, PirAB-fusion protein also exhibited injectable insecticidal activity against Spodoptera exigua larvae. The bodies of S. exigua fourth-instar larvae injected with PirAB-fusion protein turned completely black. Thus, we concluded that PirAB-fusion protein possessed similar biological activity (cytotoxicity and insecticidal activity) to PirA/PirB mixture, which would enable it to be used as an efficient agent for pest control.
Lenzmeier, B A; Nyborg, J K
The HTLV-1 promoter directs RNA polymerase II transcription of viral genomic RNA in vivo. However, it has been reported that in vitro, a unique RNA polymerase, with characteristics of RNA polymerases II and III, is capable of HTLV-1 transcription (G. Piras, F. Kashanchi, M. F. Radonovich, J. F. Duvall, and J. N. Brady, J. Virol. 68:6170-6179, 1994). To further characterize the polymerase involved in HTLV-1 transcription in vitro, runoff transcription assays were performed with a variety of extracts and RNA polymerase inhibitors. Under all in vitro reaction conditions tested, RNA polymerase II appeared to be the only polymerase capable of correct transcriptional initiation from the HTLV-1 promoter. Synthesis of the specific HTLV-1 RNA transcript showed sensitivities to the RNA polymerase inhibitors tagetitoxin and alpha-amanitin that are consistent with RNA polymerase II transcription. Together, these data indicate that in vitro, as in vivo, the HTLV-1 promoter directs transcription by RNA polymerase II. PMID:9032404
Neige, Pascal; Lapierre, Hervé; Merle, Didier
New coleoid cephalopods are described from statolith remains from the Middle Eocene (Middle Lutetian) of the Paris Basin. Fifteen fossil statoliths are identified and assigned to the Sepiidae (Sepia boletzkyi sp. nov.,? Sepia pira sp. nov.), Loliginidae (Loligo clarkei sp. nov.), and Ommastrephidae (genus indet.) families. The sediments containing these fossils indicate permanent aquatic settings in the infralittoral domain. These sediments range in age from 46 Mya to 43 Mya. Analysis of the fossil record of statoliths (from findings described here, together with a review of previously published data) indicates marked biases in our knowledge. Fossil statoliths are known from as far back as the Early Jurassic (199.3 to 190.8 Mya) but surprisingly, to the best of our knowledge, no record occurs in the Cretaceous. This is a "knowledge bias" and clearly calls for further studies. Finally, we attempt to compare findings described here with fossils previously used to constrain divergence and/or diversification ages of some coleoid subclades in molecular phylogenies. This comparison clearly indicates that the new records detailed here will challenge some estimated divergence times of coleoid cephalopod subclades.
Neige, Pascal; Lapierre, Hervé; Merle, Didier
New coleoid cephalopods are described from statolith remains from the Middle Eocene (Middle Lutetian) of the Paris Basin. Fifteen fossil statoliths are identified and assigned to the Sepiidae (Sepia boletzkyi sp. nov.,? Sepia pira sp. nov.), Loliginidae (Loligo clarkei sp. nov.), and Ommastrephidae (genus indet.) families. The sediments containing these fossils indicate permanent aquatic settings in the infralittoral domain. These sediments range in age from 46 Mya to 43 Mya. Analysis of the fossil record of statoliths (from findings described here, together with a review of previously published data) indicates marked biases in our knowledge. Fossil statoliths are known from as far back as the Early Jurassic (199.3 to 190.8 Mya) but surprisingly, to the best of our knowledge, no record occurs in the Cretaceous. This is a “knowledge bias” and clearly calls for further studies. Finally, we attempt to compare findings described here with fossils previously used to constrain divergence and/or diversification ages of some coleoid subclades in molecular phylogenies. This comparison clearly indicates that the new records detailed here will challenge some estimated divergence times of coleoid cephalopod subclades. PMID:27192490
Restrepo, Leda; Bayot, Bonny; Betancourt, Irma; Pinzón, Andres
Vibrio parahaemolyticus is a pathogenic bacteria which has been associated to the early mortality syndrome (EMS) also known as hepatopancreatic necrosis disease (AHPND) causing high mortality in shrimp farms. Pathogenic strains contain two homologous genes related to insecticidal toxin genes, PirA and PirB, these toxin genes are located on a plasmid contained within the bacteria. Genomic sequences have allowed the finding of two strains with a divergent structure related to the geographic region from where they were found. The isolates from the geographic collection of Southeast Asia and Mexico show variable regions on the plasmid genome, indicating that even though they are not alike they still conserve the toxin genes. In this paper, we report for the first time, a pathogenic V. parahaemolyticus strain in shrimp from South America that showed symptoms of AHPND. The genomic analysis revealed that this strain of V. parahaemolyticus found in South America appears to be more related to the Southeast Asia as compared to the Mexican strains. This finding is of major importance for the shrimp industry, especially in regards to the urgent need for disease control strategies to avoid large EMS outbreaks and economic loss, and to determine its dispersion in South America. The whole-genome shotgun project of V. parahaemolyticus strain Ba94C2 have been deposited at DDBJ/EMBL/GenBank under the accession PRJNA335761.
Giorgianni, C; Trimboli, K; Tanzariello, G; Fontana, G; Abbate, S; Callipari, S; Galtieri, G
The activity of roadman is not much knowed. The evalutation of the risk evidences the noise and vibrations as the principal polluter. The aim of our study is to go into the knowledge of the working risk of that job. The study was leaded on a sample of 287 roadmen. It was carried out an evalutation of noise exposure of the workers and a biologic evaluation that included: Medical check. Othorhin.laryngoiatric examination. Audiometric examination. Impedenzometric examination. The audiometric exam was conduced in a silent box and with a acoustic rest at least 16 h. The audiometric test were classified in conformity with Merluzzi-Pira-Bosio method. The phonometric evaluations point out, in roadmans examinated, hight exposure to noise with mean Lep d > 90 db. Biological evaluation, through medical check, showed data similar with that of a general group homogeneous for age and sex. Audiometric evaluation showed a noise acoustic trama in 60% of sample. Inpedenzometric exam confirmed the absence of transmissive damage in almost whole sample. In conclusion we can adfirm that noise is a strong factor for roadmen, with evident damage to exposed workers.
Franco-Martín, Manuel; Parra-Vidales, Esther; González-Palau, Fátima; Bernate-Navarro, Mara; Solis, Abdel
Introduccion. La poblacion espanola mayor de 65 anos continua creciendo, por lo que la atencion a adultos mayores se convierte en un objetivo de salud publica cada vez mas relevante. La actividad fisica esta siendo reconocida como un factor altamente protector de las funciones cognitivas en el envejecimiento, y se establece, en la actualidad, como una estrategia psicosocial prometedora para la proteccion de las facultades cognitivas. Pacientes y metodos. Mediante una busqueda bibliografica en las bases de datos PubMed, PsycINFO, Psicodoc, Scopus y SciELO, se revisaron sistematicamente los estudios relativos a la influencia del ejercicio fisico en la prevencion del deterioro cognitivo en adultos mayores sanos y en la reversion o en el mantenimiento del declive cognitivo una vez ya iniciado. Se seleccionaron y utilizaron 31 articulos como unidades de analisis. Resultados. En conjunto, estos estudios indicaron que un mayor indice de actividad fisica se relacionaria con un menor deterioro de las funciones cognitivas en adultos mayores sanos y con deterioro cognitivo ya manifiesto. Conclusiones. El ejercicio fisico constituye una estrategia psicosocial prometedora para la intervencion de adultos mayores con y sin signos de deterioro cognitivo. Serian convenientes mayores estudios que empleen una metodologia experimental, mayor homogeneidad en cuanto a los instrumentos de recogida de datos de funciones cognitivas y una profundizacion en la frecuencia e intensidad necesaria en las intervenciones.
Fröhlich, Siegmund; Sewing, Dirk
The first large-scale battery recycling facility implementing the hydrometallurgical BATENUS technology is expected to go into operation by 1996. The plant will be situated in Schönebeck/Sachsen-Anhalt, and has a projected maximum capacity of 7500 tons of spent batteries per year. The engineering is being carried out by Keramchemie GmbH and the plant will be operated by Batterierecycling Schönebeck GmbH. The BATENUS process was developed by Pira GmbH, a research institute in Stühlingen, Germany, during a period of five years. This new process combines hydrometallurgical operations in a nearly closed reagent cycle that involves electrochemical and membrane techniques. Effluent emissions are minimized to the greatest possible extent. Process validity has been proven in a series of pilot plant testings. After mechanical separation of the casing materials like ferrous and nonferrous metals, paper and plastics, the subsequent hydrometallurgical recovery yields zinc, copper, nickel and cadmium. The other products are manganese carbonate and a mixture of manganese oxide with carbon black. Mercury is immobilized by absorption on a selective ion-exchange resin. The BATENUS process is a master process for the hydrometallurgical reclamation of metals from secondary raw materials. It has found its first application in the treatment of spent consumer batteries (i.e., mixtures of zinc-carbon, alkaline manganese, lithium, nickel-cadmium cells, etc.). As a result of its modular process design, the individual steps can be modified easily and adapted to accommodate variations in the contents of the secondary raw materials. Further applications of this highly flexible technology are planned for the future.
Passos, Mauro Romero L; Varella, Renata Q; Barreto, Nero A; Garcia, Maria Luiza; Giraldo, Paulo C
Diagnosis of vaginal discharge is frequently performed in an empirical way, leading to inadequate treatment. This study tested the accuracy of a self-collection kit for microbiological study of the vaginal content. One hundred and forty-two women of Family Health Program units in Niterói and Piraí cities were enrolled in order to have their vaginal content studied. A brief explanation and a self-collection kit were provided in order to sample the vaginal content. The self-collection kit was composed of one empty plastic tube, two glass slides, a long handle cytobrush, an identification card and guideline notes. The vaginal sample was applied on the glass slides by the women and stained by Gram technique. A second sampling was done by the medical personnel. The microbiological diagnosis in a blinded analysis was made under optical microscopy. A validation diagnosis test was done taking the medical collection results as a gold standard. A total of 106 women had followed the protocol and were included in the study. Microbiological analysis was unsatisfactory in 12 cases (6 cases of self-collection material and 6 cases of medical collection). The microbiological analyses in the self-collection and in the medical collection material were respectively: bacterial vaginosis in 21.7% and 17.9%, non bacillar flora in 10.3% and 11.3%, vaginal trichomoniasis in 5.66% and 5.6%, candidiasis in 3.78% and 2.8% and a normal microbiota in 52.8% and 56.6%. The Kappa coefficient suggested a "very good correlation" of the microbiological results between the two methods of collection (K=0.7945). The self-collection kit provides samples for microbiological analysis of the vaginal microbiota as good as medical collection.
Humphrey, Mary Beth; Nakamura, Mary C
Osteoclasts require coordinated co-stimulation by several signaling pathways to initiate and regulate their cellular differentiation. Receptor activator for NF-κB ligand (RANKL or TNFSF11), a tumor necrosis factor (TNF) superfamily member, is the master cytokine required for osteoclastogenesis with essential co-stimulatory signals mediated by immunoreceptor tyrosine-based activation motif (ITAM)-signaling adaptors, DNAX-associated protein 12 kDa size (DAP12) and FcεRI gamma chain (FcRγ). The ITAM-signaling adaptors do not have an extracellular ligand-binding domain and, therefore, must pair with ligand-binding immunoreceptors to interact with their extracellular environment. DAP12 pairs with a number of different immunoreceptors including triggering receptor expressed on myeloid cells 2 (TREM2), myeloid DAP12-associated lectin (MDL-1), and sialic acid-binding immunoglobulin-type lectin 15 (Siglec-15); while FcRγ pairs with a different set of receptors including osteoclast-specific activating receptor (OSCAR), paired immunoglobulin receptor A (PIR-A), and Fc receptors. The ligands for many of these receptors in the bone microenvironment remain unknown. Here, we will review immunoreceptors known to pair with either DAP12 or FcRγ that have been shown to regulate osteoclastogenesis. Co-stimulation and the effects of ITAM-signaling have turned out to be complex, and now include paradoxical findings that ITAM-signaling adaptor-associated receptors can inhibit osteoclastogenesis and immunoreceptor tyrosine-based inhibitory motif (ITIM) receptors can promote osteoclastogenesis. Thus, co-stimulation of osteoclastogenesis continues to reveal additional complexities that are important in the regulatory mechanisms that seek to maintain bone homeostasis.
Qi, Yanhong; Yu, Yang; Li, Kaisheng; Chen, Zhihong; Liu, Yuanchang; Zhang, Hongmei
Introducción: En los últimos años, la obesidad se ha convertido en el principal problema de salud pública a nivel mundial. La tasa de detección de obesidad infantil está creciendo alarmantemente entre los niños y los adolescentes en China, cuya población de 120 millones se sitúa en el rango de obesidad. Se calculó que el 8% de los niños son obesos. Métodos: Se recopilaron publicaciones entre 2009 y 2014 sobre la prevalencia de obesidad entre los alumnos de primaria en China de PubMed, textos completos de bases de datos de periódicos chinos online de VIP, CNKI y Wanfang. Se empleó software de meta-análisis para resumir y analizar las tasas de detección incluidas en la bibliografía previamente obtenida. Resultados: Después de la evaluación de la calidad de los artículos, finalmente incluimos 18 artículos en nuestro estudio, y los tamaños de la muestra total en la investigación sobre obesidad fueron 247547, donde 26466 presentaron obesidad. Los resultados de los meta-análisis mostraron que la prevalencia de la obesidad en los alumnos de primaria es del 10,0% (IC del 95%: 7,2% - 13,5%). Conclusión: Nuestros resultados indican que la prevalencia de obesidad en China todavía supone un problema preocupante, dado que la situación será peor si no se toman medidas eficaces y prácticas.
Riveros, Efraín; Amado, Paula; Zambrano, Paula
Introducción: La seguridad de paciente en áreas hospitalarias críticas ha sido tema de diversas investigaciones. En este sentido, en Colombia está creciendo la conciencia acerca de la gestión de eventos adversos. Este estudio explora la primera fase de dicha gestión: el reporte en la unidad de cuidado intensivo de una institución privada prototipo. Objetivo: Evaluar el reporte de eventos adversos en una clínica colombiana de tercer nivel, a través de la estimación de frecuencias y categorías de los reportes. Material y Métodos: Se hizo un estudio descriptivo sobre la tasa de reporte de eventos adversos en la Clínica de los Andes de Tunja, durante 2011, a través de la estimación de frecuencias y categorías de los reportes.La tasa de reporte se definió como el número de reportes por 1000 días cama. Reportamos las categorías de los eventos adversos comomayores y menores. Resultados: Se registraron 383 admisiones a la UCI, correspondientes a 2374 días paciente. Se presentaron un total de 54 eventos adversos reportados (tasa de 22,74 por 1000 días paciente). El tipo de evento más frecuente fue el relacionado con vías intravenosas (55%)(IC 95% 50,1-58,3), seguido por desviación de procesos (22%)(IC 95% 18,2-26,1). Conclusiones: Se encontróuna tasa de reporte de eventos adversos en el rango superior de lo publicado en la literatura. Posiblemente exista aúnuna proporción no cuantificada de subregistro, que puede atribuirse principalmente a factores culturales.
Hegg, Jens C; Giarrizzo, Tommaso; Kennedy, Brian P
Animal migrations provide important ecological functions and can allow for increased biodiversity through habitat and niche diversification. However, aquatic migrations in general, and those of the world's largest fish in particular, are imperiled worldwide and are often poorly understood. Several species of large Amazonian catfish carry out some of the longest freshwater fish migrations in the world, travelling from the Amazon River estuary to the Andes foothills. These species are important apex predators in the main stem rivers of the Amazon Basin and make up the region's largest fishery. They are also the only species to utilize the entire Amazon Basin to complete their life cycle. Studies indicate both that the fisheries may be declining due to overfishing, and that the proposed and completed dams in their upstream range threaten spawning migrations. Despite this, surprisingly little is known about the details of these species' migrations, or their life history. Otolith microchemistry has been an effective method for quantifying and reconstructing fish migrations worldwide across multiple spatial scales and may provide a powerful tool to understand the movements of Amazonian migratory catfish. Our objective was to describe the migratory behaviors of the three most populous and commercially important migratory catfish species, Dourada (Brachyplatystoma rousseauxii), Piramutaba (Brachyplatystoma vaillantii), and Piraíba (Brachyplatystoma filamentosum). We collected fish from the mouth of the Amazon River and the Central Amazon and used strontium isotope signatures ((87)Sr/(86)Sr) recorded in their otoliths to determine the location of early rearing and subsequent. Fish location was determined through discriminant function classification, using water chemistry data from the literature as a training set. Where water chemistry data was unavailable, we successfully in predicted (87)Sr/(86)Sr isotope values using a regression-based approach that related the geology
McTavish, H; Sayavedra-Soto, L A; Arp, D J
Mutants in which conserved cysteines 294, 297 or 64 and 65 of the Azotobacter vinelandii hydrogenase small subunit were replaced by serines were studied. Cysteines 294 and 297 are homologous to cysteines 246 and 249 of the Desulfovibrio gigas hydrogenase, and these cysteines are ligands to the [3Fe-4S] clusters (A. Volbeda, M.-H. Charon, C. Piras, E. C. Hatchikian, M. Frey, and J. C. Fontecilla-Camps, Nature (London) 373:580-587, 1995). Cysteine 65 is homologous to cysteine 20 of the D. gigas hydrogenase, and this cysteine is a ligand to the proximal [4Fe-4S] cluster. All three mutants retained some hydrogenase activity. All three mutants studied had H2 oxidation-to-H2 evolution activity ratios with whole cells of approximately 1.5, compared with 46 for the wild type. The changes preferentially deplete H2 oxidation activity, while having less effect on evolution. The K64,65C-->S hydrogenase was partially purified and had a specific activity for the evolution reaction that was 22% that of the wild type, while the oxidation-specific activity was 2% that of the wild type. Because cysteine 65 provides a ligand to the proximal [4Fe-4S] cluster, this cluster can be altered without entirely eliminating enzyme activity. Likewise, the detection of H2 evolution and H2 oxidation activities with whole cells and membranes of the K294C-->S and K297C-->S mutants indicates that the [3Fe-4S] cluster can also be altered or possibly eliminated without entirely eliminating enzyme activity. Membranes with K294C-->S or K297C-->S hydrogenase were uninhibited by O2 in H2 oxidation and uninhibited by H2 in H2 evolution. Wild-type membranes and membranes with K64,65C-->S hydrogenase were both sensitive to these inhibitors. These data indicate that the [3Fe-4S] cluster controls the reversible inhibition of hydrogenase activity by O2 or H2. PMID:7608067
Hegg, Jens C.; Giarrizzo, Tommaso; Kennedy, Brian P.
Animal migrations provide important ecological functions and can allow for increased biodiversity through habitat and niche diversification. However, aquatic migrations in general, and those of the world’s largest fish in particular, are imperiled worldwide and are often poorly understood. Several species of large Amazonian catfish carry out some of the longest freshwater fish migrations in the world, travelling from the Amazon River estuary to the Andes foothills. These species are important apex predators in the main stem rivers of the Amazon Basin and make up the region’s largest fishery. They are also the only species to utilize the entire Amazon Basin to complete their life cycle. Studies indicate both that the fisheries may be declining due to overfishing, and that the proposed and completed dams in their upstream range threaten spawning migrations. Despite this, surprisingly little is known about the details of these species’ migrations, or their life history. Otolith microchemistry has been an effective method for quantifying and reconstructing fish migrations worldwide across multiple spatial scales and may provide a powerful tool to understand the movements of Amazonian migratory catfish. Our objective was to describe the migratory behaviors of the three most populous and commercially important migratory catfish species, Dourada (Brachyplatystoma rousseauxii), Piramutaba (Brachyplatystoma vaillantii), and Piraíba (Brachyplatystoma filamentosum). We collected fish from the mouth of the Amazon River and the Central Amazon and used strontium isotope signatures (87Sr/86Sr) recorded in their otoliths to determine the location of early rearing and subsequent. Fish location was determined through discriminant function classification, using water chemistry data from the literature as a training set. Where water chemistry data was unavailable, we successfully in predicted 87Sr/86Sr isotope values using a regression-based approach that related the geology of
Rodríguez-Rodríguez, Laura; Jiménez-Sánchez, Montserrat; Domínguez-Cuesta, María Jose; Rinterknecht, Vincent; Pallàs, Raimon; Braucher, Régis; Bourlès, Didier; Valero-Garcés, Blas
., 2011. Last deglaciation in northwestern Spain: New chronological and geomorphologic evidence from the Sanabria region. Geomorphology 135, 48-65. Palacios, D., Andrés, N., Úbeda, J., Alcalá, J., Marcos, J., Vázquez-Selem, L., 2012. The importance of poligenic moraines in the paleoclimatic interpretation from cosmogenic dating. Geophysical Research Abstracts 14, EGU2012-3759-1. Pérez-Alberti, A., Valcárcel-Díaz, M., Martini, I.P., Pascucci, V., Andrucci, S., 2011. Upper Pleistocene glacial valley-junction sediments at Pias, Trevinca Mountains, NW Spain. In: Martini, I.P., French, H.M., Pérez-Alberti, A. (Eds.), Ice-Marginal and Periglacial Processes and Sediments. Geological Society (London) Special Publication 354, pp. 93-110. Research funded by the projects LIMNOCLIBER (REN2003-09130-C02-02), IBERLIMNO (CGL2005-20236-E/CLI), LIMNOCAL (CGL2006-13327-C04-01) and GRACCIE (CSD2007-00067) of the Spanish Inter-Ministry Commission of Science and Technology (CICYT). Additional funding was provided by the Fundación Patrimonio Natural de Castilla y León through the project "La investigacion en el Lago de Sanabria dentro del proyecto CALIBRE: perspectivas y posibilidades", and by the projects Consolider Ingenio 2006 (CSD2006-0041, Topo-Iberia), 2003 PIRA 00256, HF02.4, and RISKNAT (2009SGR520). L. Rodríguez-Rodríguez has developed her research under a Severo Ochoa Programme fellowship (FICYT- Asturias).