Eighteen Years of Molecular Genotyping the Hemophilia Inversion Hotspot: From Southern Blot to Inverse Shifting-PCR

PubMed Central

Rossetti, Liliana C.; Radic, Claudia P.; Abelleyro, Miguel M.; Larripa, Irene B.; De Brasi, Carlos D.

2011-01-01

The factor VIII gene (F8) intron 22 inversion (Inv22) is a paradigmatic duplicon-mediated rearrangement, found in about one half of patients with severe hemophilia A worldwide. The identification of this prevalent cause of hemophilia was delayed for nine years after the F8 characterization in 1984. The aim of this review is to present the wide diversity of practical approaches that have been developed for genotyping the Inv22 (and related int22h rearrangements) since discovery in 1993. The sequence— Southern blot, long distance-PCR and inverse shifting-PCR—for Inv22 genotyping is an interesting example of scientific ingenuity and evolution in order to resolve challenging molecular diagnostic problems. PMID:22072947

Retrospective Study of Hemoparasites in Cattle in Southern Italy by Reverse Line Blot Hybridization

PubMed Central

CECI, Luigi; IARUSSI, Fabrizio; GRECO, Beatrice; LACINIO, Rosanna; FORNELLI, Stefania; CARELLI, Grazia

2014-01-01

ABSTRACT Tick-borne diseases are widespread in tropical and temperate regions and are responsible for important economic losses in those areas. In order to assess the presence and prevalence of various pathogens in southern Italy, we retrospectively analyzed cattle blood samples collected for a previous study in 2000 using reverse line blot (RLB) hybridization. The study had been carried out in three regions of southern Italy on 1,500 randomly selected and apparently healthy adult cattle. RLB showed that 43.7% of the cattle were positive for nine different species of hemoparasites with either a single infection or a mixed infection. Theileria buffeli was the most common species found, being present in 27.3% of the animals, followed by Anaplasma marginale in 18.1%, Anaplasma centrale in 13.8%, Babesia bigemina and Anaplasma bovis in 4.2%, Anaplasma phagocytophilum in 1.7%, Babesia bovis in 1.6%, Babesia major in 0.2% and Babesia divergens in 0.1%. Complete blood counts showed different degrees of anemia in 363 animals (24.2%) and of these, 169 were RLB-positive for at least one pathogen. Among the ticks that were collected from the cattle, the following species were identified: Rhipicephalus bursa, Ixodes ricinus, Hyalomma marginatum, Boophilus annulatus, Dermacentor marginatus and Haemaphysalis (sulcata, parva, inermis and punctata). The results obtained confirmed the spread of endemic tick-borne pathogens in the regions studied. PMID:24614604

Retrospective study of hemoparasites in cattle in southern Italy by reverse line blot hybridization.

PubMed

Ceci, Luigi; Iarussi, Fabrizio; Greco, Beatrice; Lacinio, Rosanna; Fornelli, Stefania; Carelli, Grazia

2014-06-01

Tick-borne diseases are widespread in tropical and temperate regions and are responsible for important economic losses in those areas. In order to assess the presence and prevalence of various pathogens in southern Italy, we retrospectively analyzed cattle blood samples collected for a previous study in 2000 using reverse line blot (RLB) hybridization. The study had been carried out in three regions of southern Italy on 1,500 randomly selected and apparently healthy adult cattle. RLB showed that 43.7% of the cattle were positive for nine different species of hemoparasites with either a single infection or a mixed infection. Theileria buffeli was the most common species found, being present in 27.3% of the animals, followed by Anaplasma marginale in 18.1%, Anaplasma centrale in 13.8%, Babesia bigemina and Anaplasma bovis in 4.2%, Anaplasma phagocytophilum in 1.7%, Babesia bovis in 1.6%, Babesia major in 0.2% and Babesia divergens in 0.1%. Complete blood counts showed different degrees of anemia in 363 animals (24.2%) and of these, 169 were RLB-positive for at least one pathogen. Among the ticks that were collected from the cattle, the following species were identified: Rhipicephalus bursa, Ixodes ricinus, Hyalomma marginatum, Boophilus annulatus, Dermacentor marginatus and Haemaphysalis (sulcata, parva, inermis and punctata). The results obtained confirmed the spread of endemic tick-borne pathogens in the regions studied.

Western Blotting of the Endocannabinoid System.

PubMed

Wager-Miller, Jim; Mackie, Ken

2016-01-01

Measuring expression levels of G protein-coupled receptors (GPCRs) is an important step for understanding the distribution, function, and regulation of these receptors. A common approach for detecting proteins from complex biological systems is Western blotting. In this chapter, we describe a general approach to Western blotting protein components of the endocannabinoid system using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nitrocellulose membranes, with a focus on detecting type 1 cannabinoid (CB1) receptors. When this technique is carefully used, specifically with validation of the primary antibodies, it can provide quantitative information on protein expression levels. Additional information can also be inferred from Western blotting such as potential posttranslational modifications that can be further evaluated by specific analytical techniques.

Application of FTA sample collection and DNA purification system on the determination of CTG trinucleotide repeat size by PCR-based Southern blotting.

PubMed

Hsiao, K M; Lin, H M; Pan, H; Li, T C; Chen, S S; Jou, S B; Chiu, Y L; Wu, M F; Lin, C C; Li, S Y

1999-01-01

Myotonic dystrophy (DM) is caused by a CTG trinucleotide expansion mutation at exon 15 of the myotonic dystrophy protein kinase gene. The clinical severity of this disease correlates with the length of the CTG trinucleotide repeats. Determination of the CTG repeat length has been primarily relied on by Southern blot analysis of restriction enzyme-digested genomic DNA. The development of PCR-based Southern blotting methodology provides a much more sensitive and simpler protocol for DM diagnosis. However, the quality of the template and the high (G+C) ratio of the amplified region hamper the use of PCR on the diagnosis of DM. A modified PCR protocol to amplify different lengths of CTG repeat region using various concentrations of 7deaza-dGTP has been reported (1). Here we describe a procedure including sample collection, DNA purification, and PCR analysis of CTG repeat length without using 7-deaza-dGTP. This protocol is very sensitive and convenient because only a small number of nucleate cells are needed for detection of CTG expansion. Therefore, it could be very useful in clinical and prenatal diagnosis as well as in prevalence study of DM.

Western blotting.

PubMed

Kurien, Biji T; Scofield, R Hal

2006-04-01

Western blotting (protein blotting or immunoblotting) is a powerful and important procedure for the immunodetection of proteins post-electrophoresis, particularly proteins that are of low abundance. Since the inception of the protocol for protein transfer from an electrophoresed gel to a membrane in 1979, protein blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer. This review describes the various procedures that have been used to transfer proteins from a gel to a membrane based on the principles of simple diffusion, vacuum-assisted solvent flow and electrophoretic elution. Finally, a brief description of methods generally used to detect antigens on blots is also described.

Western Blot of Stained Proteins from Dried Polyacrylamide Gels

NASA Technical Reports Server (NTRS)

Gruber, Claudia; Stan-Lotter, Helga

1996-01-01

Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.

The combination of quantitative PCR and western blot detecting CP4-EPSPS component in Roundup Ready soy plant tissues and commercial soy-related foodstuffs.

PubMed

Xiao, Xiao; Wu, Honghong; Zhou, Xinghu; Xu, Sheng; He, Jian; Shen, Wenbiao; Zhou, Guanghong; Huang, Ming

2012-06-01

With the widespread use of Roundup Ready soy (event 40-3-2) (RRS), the comprehensive detection of genetically modified component in foodstuffs is of significant interest, but few protein-based approaches have been found useful in processed foods. In this report, the combination of quantitative PCR (qPCR) and western blot was used to detect cp4-epsps gene and its protein product in different RRS plant tissues and commercial soy-containing foodstuffs. The foods included those of plant origin produced by different processing procedures and also some products containing both meat and plant protein concentrates. The validity of the 2 methods was confirmed first. We also showed that the CP4-EPSPS protein existed in different RRS plant tissues. In certain cases, the results from the western blot and the qPCR were not consistent. To be specific, at least 2 degraded fragments of CP4-EPSPS protein (35.5 and 24.6 kDa) were observed. For dried bean curd crust and deep-fried bean curd, a degraded protein fragment with the size of 24.6 kDa appeared, while cp4-epsps gene could not be traced by qPCR. In contrast, we found a signal of cp4-epsps DNA in 3 foodstuffs, including soy-containing ham cutlet product, meat ball, and sausage by qPCR, while CP4-EPSPS protein could not be detected by western blot in such samples. Our study therefore concluded that the combination of DNA- and protein-based methods would compensate each other, thus resulting in a more comprehensive detection from nucleic acid and protein levels. The combination of quantitative PCR (qPCR) and western blot was used to detect cp4-epsps gene and its protein product in different Roundup Ready soy (event 40-3-2) plant tissues and commercial soy-containing foodstuffs. The foods included those of plant origin produced by different processing procedures and also some products containing a combination of both meat and plant protein concentrates. This study indicated that the combination of DNA- and protein-based methods

The western blot

USDA-ARS?s Scientific Manuscript database

Western blotting is a technique that involves the separation of proteins by gel electrophoresis, their blotting or transfer to a membrane, and selective immunodetection of an immobilized antigen. This is an important and routine method for protein analysis that depends on the specificity of antibod...

Western blotting using capillary electrophoresis.

PubMed

Anderson, Gwendolyn J; M Cipolla, Cynthia; Kennedy, Robert T

2011-02-15

A microscale Western blotting system based on separating sodium-dodecyl sulfate protein complexes by capillary gel electrophoresis followed by deposition onto a blotting membrane for immunoassay is described. In the system, the separation capillary is grounded through a sheath capillary to a mobile X-Y translation stage which moves a blotting membrane past the capillary outlet for protein deposition. The blotting membrane is moistened with a methanol and buffer mixture to facilitate protein adsorption. Although discrete protein zones could be detected, bands were broadened by ∼1.7-fold by transfer to membrane. A complete Western blot for lysozyme was completed in about one hour with 50 pg mass detection limit from low microgram per milliliter samples. These results demonstrate substantial reduction in time requirements and improvement in mass sensitivity compared to conventional Western blots. Western blotting using capillary electrophoresis shows promise to analyze low volume samples with reduced reagents and time, while retaining the information content of a typical Western blot.

Lipid-binding analysis using a fat blot assay.

PubMed

Munnik, Teun; Wierzchowiecka, Magdalena

2013-01-01

Protein-lipid interactions play an important role in lipid metabolism, membrane trafficking and cell -signaling by regulating protein localization, activation, and function. The Fat Blot assay is a relatively simple and inexpensive method to examine these interactions using nitrocellulose membrane-immobilized lipids. The assay is adapted from the method by Dowler et al. (Sci STKE 129:pl6, 2002) and provides qualitative and quantitative information on the relative affinity with which a protein binds to a particular lipid. To perform a Fat Blot assay, serial dilutions of different phospholipids are spotted onto a nitrocellulose membrane. These membranes are then incubated with a lipid-binding protein possessing a GST (or other epitope) tag. The membranes are washed and the protein, which is bound to the membrane by virtue of its interaction with the lipid's head group, is detected by immunoblotting with an antibody against GST (or other epitope). The procedure only requires a few micrograms of protein and is quick, simple and cheap to perform.

Value of molecular monitoring during the treatment of chronic myeloid leukemia: a Cancer and Leukemia Group B study.

PubMed

Stock, W; Westbrook, C A; Peterson, B; Arthur, D C; Szatrowski, T P; Silver, R T; Sher, D A; Wu, D; Le Beau, M M; Schiffer, C A; Bloomfield, C D

1997-01-01

Disappearance of the Philadelphia chromosome during treatment for chronic myeloid leukemia (CML) has become an important therapeutic end point. To determine the additional value of molecular monitoring during treatment for CML, we performed a prospective, sequential analysis using quantitative Southern blot monitoring of BCR gene rearrangements of blood and marrow samples from Cancer and Leukemia Group B (CALGB) study 8761. Sixty-four previously untreated adults with chronic-phase CML who were enrolled onto CALGB 8761, a molecular-monitoring companion study to a treatment study for adults with chronic-phase CML (CALGB 9013). Treatment consisted of repetitive cycles of interferon alfa and low-dose subcutaneous cytarabine. Blood and marrow Southern blot quantitation of BCR gene rearrangements was compared with marrow cytogenetic analysis before the initiation of treatment and of specified points during therapy. Reverse-transcriptase polymerase chain reaction (RT-PCR) analysis was performed to detect residual disease in patients who achieved a complete response by Southern blot or cytogenetic analysis. Quantitative molecular monitoring by Southern blot analysis of blood samples was found to be equivalent to marrow monitoring at all time points. Twelve of 62 (19%) follow-up samples studied by Southern blot analysis had a complete loss of BCR gene rearrangement in matched marrow and blood specimens. Southern blot monitoring of blood samples was also found to be highly correlated to marrow cytogenetic evaluation at all points, although there were four discordant cases in which Southern blot analysis of blood showed no BCR gene rearrangement, yet demonstrated from 12% to 20% Philadelphia chromosome-positive metaphase cells in the marrow. RT-PCR analysis detected residual disease in five of six patients in whom no malignant cells were detected using Southern blot or cytogenetic analyses. Quantitative Southern blot analysis of blood samples may be substituted for bone

Introduction to protein blotting.

PubMed

Kurien, Biji T; Scofield, R Hal

2009-01-01

Protein blotting is a powerful and important procedure for the immunodetection of proteins following electrophoresis, particularly proteins that are of low abundance. Since the inception of the protocol for protein transfer from an electrophoresed gel to a membrane in 1979, protein blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer.

Western blotting: an introduction.

PubMed

Kurien, Biji T; Scofield, R Hal

2015-01-01

Western blotting is an important procedure for the immunodetection of proteins, particularly proteins that are of low abundance. This process involves the transfer of protein patterns from gel to microporous membrane. Electrophoretic as well as non-electrophoretic transfer of proteins to membranes was first described in 1979. Protein blotting has evolved greatly since the inception of this protocol, allowing protein transfer to be accomplished in a variety of ways.

A new background subtraction method for Western blot densitometry band quantification through image analysis software.

PubMed

Gallo-Oller, Gabriel; Ordoñez, Raquel; Dotor, Javier

2018-06-01

Since its first description, Western blot has been widely used in molecular labs. It constitutes a multistep method that allows the detection and/or quantification of proteins from simple to complex protein mixtures. Western blot quantification method constitutes a critical step in order to obtain accurate and reproducible results. Due to the technical knowledge required for densitometry analysis together with the resources availability, standard office scanners are often used for the imaging acquisition of developed Western blot films. Furthermore, the use of semi-quantitative software as ImageJ (Java-based image-processing and analysis software) is clearly increasing in different scientific fields. In this work, we describe the use of office scanner coupled with the ImageJ software together with a new image background subtraction method for accurate Western blot quantification. The proposed method represents an affordable, accurate and reproducible approximation that could be used in the presence of limited resources availability. Copyright © 2018 Elsevier B.V. All rights reserved.

Detection of Proteins on Blot Membranes

PubMed Central

Goldman, Aaron; Harper, Sandra; Speicher, David W.

2017-01-01

Staining of blot membranes enables the visualization of bound proteins. Proteins are usually transferred to blot membranes by electroblotting, by direct spotting of protein solutions, or by contact blots. Staining allows the efficiency of transfer to the membrane to be monitored. This unit describes protocols for staining proteins after electroblotting from polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. The same methods can be used if proteins are directly spotted, either manually or using robotics. Protocols are included for seven general protein stains (amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, India ink, and MemCode) and three fluorescent protein stains (fluorescamine, IAEDANS, and SYPRO Ruby). Also included is an in-depth discussion of the different blot membrane types and the compatibility of different protein stains with downstream applications, such as immunoblotting or N-terminal Edman sequencing. PMID:27801518

Detection of Proteins on Blot Membranes.

PubMed

Goldman, Aaron; Harper, Sandra; Speicher, David W

2016-11-01

Staining of blot membranes enables the visualization of bound proteins. Proteins are usually transferred to blot membranes by electroblotting, by direct spotting of protein solutions, or by contact blots. Staining allows the efficiency of transfer to the membrane to be monitored. This unit describes protocols for staining proteins after electroblotting from polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. The same methods can be used if proteins are directly spotted, either manually or using robotics. Protocols are included for seven general protein stains (amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, India ink, and MemCode) and three fluorescent protein stains (fluorescamine, IAEDANS, and SYPRO Ruby). Also included is an in-depth discussion of the different blot membrane types and the compatibility of different protein stains with downstream applications, such as immunoblotting or N-terminal Edman sequencing. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

Detection of proteins on blot transfer membranes.

PubMed

Sasse, Joachim; Gallagher, Sean R

2003-11-01

In the basic and alternate protocols of this unit, proteins are stained after electroblotting from polyacrylamide gels to blot transfer membranes. If the samples of interest are electrophoresed in duplicate and transferred to a blot transfer membrane, half of the membrane can be stained to determine the efficiency of transfer to the membrane and the other half can be used for immunoblotting (i.e., western blotting). Detection limits of each staining method are given along with a list of compatible blot transfer membranes and gels. A support protocol describes a method for alkali treatment that enhances subsequent staining of bound proteins.

Nonenzymatic chemiluminescent detection and quantitation of total protein on Western and slot blots allowing subsequent immunodetection and sequencing.

PubMed

Alba, F J; Daban, J R

1997-10-01

We have studied the light emission efficiency of proteins labeled with different fluorescent dyes chemically excited by the bis(2,4,6-trichlorophenyl)oxalate (TCPO)-H2O2 reaction. Using this peroxyoxalate chemiluminescence system, the best results were obtained with proteins covalently labeled with 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF). Blotted proteins on polyvinylidene difluoride (PVDF) membranes can be labeled rapidly with MDPF. Our results demonstrate that energy from the excited intermediate produced in the TCPO-H2O2 reaction can be efficiently transferred to MDPF-labeled proteins in solution and on PVDF membranes. Although this nonenzymatic chemiluminescent system produces a background emission that reduces the sensitivity, the method developed in this work allows detection of 5 ng of protein in blots after 5 min exposure to X-ray film. Chemiluminescence of MDPF-labeled proteins on Western and slot blots may also be detected and quantified using a charge-coupled device (CCD) camera or a storage phosphor imaging system. This chemiluminescent method allows the staining of the total electrophoretic pattern but does not preclude further N-terminal sequencing and immunodetection of specific bands.

Protein purification and analysis: next generation Western blotting techniques.

PubMed

Mishra, Manish; Tiwari, Shuchita; Gomes, Aldrin V

2017-11-01

Western blotting is one of the most commonly used techniques in molecular biology and proteomics. Since western blotting is a multistep protocol, variations and errors can occur at any step reducing the reliability and reproducibility of this technique. Recent reports suggest that a few key steps, such as the sample preparation method, the amount and source of primary antibody used, as well as the normalization method utilized, are critical for reproducible western blot results. Areas covered: In this review, improvements in different areas of western blotting, including protein transfer and antibody validation, are summarized. The review discusses the most advanced western blotting techniques available and highlights the relationship between next generation western blotting techniques and its clinical relevance. Expert commentary: Over the last decade significant improvements have been made in creating more sensitive, automated, and advanced techniques by optimizing various aspects of the western blot protocol. New methods such as single cell-resolution western blot, capillary electrophoresis, DigiWest, automated microfluid western blotting and microchip electrophoresis have all been developed to reduce potential problems associated with the western blotting technique. Innovative developments in instrumentation and increased sensitivity for western blots offer novel possibilities for increasing the clinical implications of western blot.

Single cell–resolution western blotting

PubMed Central

Kang, Chi-Chih; Yamauchi, Kevin A; Vlassakis, Julea; Sinkala, Elly; Duncombe, Todd A; Herr, Amy E

2017-01-01

This protocol describes how to perform western blotting on individual cells to measure cell-to-cell variation in protein expression levels and protein state. like conventional western blotting, single-cell western blotting (scWB) is particularly useful for protein targets that lack selective antibodies (e.g., isoforms) and in cases in which background signal from intact cells is confounding. scWB is performed on a microdevice that comprises an array of microwells molded in a thin layer of a polyacrylamide gel (PAG). the gel layer functions as both a molecular sieving matrix during PAGE and a blotting scaffold during immunoprobing. scWB involves five main stages: (i) gravity settling of cells into microwells; (ii) chemical lysis of cells in each microwell; (iii) PAGE of each single-cell lysate; (iv) exposure of the gel to UV light to blot (immobilize) proteins to the gel matrix; and (v) in-gel immunoprobing of immobilized proteins. Multiplexing can be achieved by probing with antibody cocktails and using antibody stripping/reprobing techniques, enabling detection of 10+ proteins in each cell. We also describe microdevice fabrication for both uniform and pore-gradient microgels. to extend in-gel immunoprobing to gels of small pore size, we describe an optional gel de-cross-linking protocol for more effective introduction of antibodies into the gel layer. once the microdevice has been fabricated, the assay can be completed in 4–6 h by microfluidic novices and it generates high-selectivity, multiplexed data from single cells. the technique is relevant when direct measurement of proteins in single cells is needed, with applications spanning the fundamental biosciences to applied biomedicine. PMID:27466711

[Better performance of Western blotting: quick vs slow protein transfer, blotting membranes and the visualization methods].

PubMed

Kong, Ling-Quan; Pu, Ying-Hui; Ma, Shi-Kun

2008-01-01

To study how the choices of the quick vs slow protein transfer, the blotting membranes and the visualization methods influence the performance of Western blotting. The cellular proteins were abstracted from human breast cell line MDA-MB-231 for analysis with Western blotting using quick (2 h) and slow (overnight) protein transfer, different blotting membranes (nitrocellulose, PVDF and nylon membranes) and different visualization methods (ECL and DAB). In Western blotting with slow and quick protein transfer, the prestained marker presented more distinct bands on nitrocellulose membrane than on the nylon and PVDF membranes, and the latter also showed clear bands on the back of the membrane to very likely cause confusion, which did not occur with nitrocellulose membrane. PVDF membrane allowed slightly clearer visualization of the proteins with DAB method as compared with nitrocellulose and nylon membranes, and on the latter two membranes, quick protein transfer was likely to result in somehow irregular bands in comparison with slow protein transfer. With slow protein transfer and chemiluminescence for visualization, all the 3 membranes showed clear background, while with quick protein transfer, nylon membrane gave rise to obvious background noise but the other two membranes did not. Different membranes should be selected for immunoblotting according to the actual needs of the experiment. Slow transfer of the proteins onto the membranes often has better effect than quick transfer, and enhanced chemiluminescence is superior to DAB for protein visualization and allows highly specific and sensitive analysis of the protein expressions.

Western blotting revisited: critical perusal of underappreciated technical issues.

PubMed

Gorr, Thomas A; Vogel, Johannes

2015-04-01

The most commonly used semiquantitative analysis of protein expression still employs protein separation by denaturing SDS-PAGE with subsequent Western blotting and quantification of the resulting ODs of bands visualized with specific antibodies. However, many questions regarding this procedure are usually ignored, although still in need of answering: Does isolation or separation procedure harm the integrity or affect modifications (e.g., phosphorylation) of the protein of interest? Does denaturation reduce binding of antibodies used for detection? Should denaturation be performed or should a native gel be run? How can artificial degradations or aggregations be distinguished from biological relevant ones? If the antibody detects multiple bands (which is not uncommon), which one(s) should be taken into account for quantification and why? Which loading control protein should be chosen and is it really "housekeeping" and how can this be verified? Is the image acquisition system linear and does it come with a sufficient dynamic range? How to account and control for background staining? This article is intended to address these questions and raise the readers awareness to possible Western blot alternatives in the attempt of minimizing possible pitfalls that might loom anywhere from protein isolation to acquisition of final quantitative data. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Novel Methylation PCR that Offers Standardized Determination of FMR1 Methylation and CGG Repeat Length without Southern Blot Analysis

PubMed Central

Grasso, Marina; Boon, Elles M.J.; Filipovic-Sadic, Stela; van Bunderen, Patrick A.; Gennaro, Elena; Cao, Ru; Latham, Gary J.; Hadd, Andrew G.; Coviello, Domenico A.

2015-01-01

Fragile X syndrome and associated disorders are characterized by the number of CGG repeats and methylation status of the FMR1 gene for which Southern blot (SB) historically has been required for analysis. This study describes a simple PCR-only workflow (mPCR) to replace SB analysis, that incorporates novel procedural controls, treatment of the DNA in separate control and methylation-sensitive restriction endonuclease reactions, amplification with labeled primers, and two-color amplicon sizing by capillary electrophoresis. mPCR was evaluated in two independent laboratories with 76 residual clinical samples that represented typical and challenging fragile X alleles in both males and females. mPCR enabled superior size resolution and analytical sensitivity for size and methylation mosaicism compared to SB. Full mutation mosaicism was detected down to 1% in a background of 99% normal allele with 50- to 100-fold less DNA than required for SB. A low level of full mutation mosaicism in one sample was detected using mPCR but not observed using SB. Overall, the sensitivity for detection of full mutation alleles was 100% (95% CI: 89%–100%) with an accuracy of 99% (95% CI: 93%–100%). mPCR analysis of DNA from individuals with Klinefelter and Turner syndromes, and DNA from sperm and blood, were consistent with SB. As such, mPCR enables accurate, sensitive, and standardized methods of FMR1 analysis that can harmonize results across different laboratories. PMID:24177047

[Scientific Evaluation of Crude Drugs and Kampo Medicines Using the Eastern Blotting Method and Its Application to Biological Metabolic Studies].

PubMed

Morinaga, Osamu

2018-01-01

　The scientific evaluation of crude drugs and kampo medicines (KMs) was demonstrated using the eastern blotting method with monoclonal antibodies (MAbs) against bioactive natural compounds. Scutellariae radix is one of the most important crude drugs used in KMs. Part of its pharmaceutical properties is due to the flavone glycoside baicalin (BI). A quantitative analysis method based on eastern blotting was developed for BI using an anti-BI MAb. A rapid, simple, sensitive, specific analytical system was subsequently established for BI with the eastern blotting technique using dot-blot and chemiluminescent methods. This system was useful as a high-throughput analytical method for the determination of BI in KMs as well as HPLC and enzyme-linked immunosorbent assay systems. Furthermore, an eastern blotting method was applied to the biological metabolic study of glycyrrhizic acid (GL), the major active constituent of licorice, for investigation of metabolites of GL such as 3-monoglucuronyl-glycyrrhetinic acid (3MGA) because licorice causes pseudoaldosteronism as a side effect. This approach may make it possible to determine the pathogenic agents of licorice-induced pseudoaldosteronism.

The Dot Blot ELISA.

ERIC Educational Resources Information Center

Gerbig, Donald G., Jr.; Fenk, Christopher J.; Goodhart, Amy S.

2000-01-01

Uses two laboratory techniques, Enzyme Linked Immunosorbent Assay (ELISA) and Western Blot, to demonstrate antibody-antigen binding concepts. Includes a list of required materials and directions for the procedure, and makes suggestions for classroom applications. (Contains 13 references.) (YDS)

Multiplexed Western Blotting Using Microchip Electrophoresis.

PubMed

Jin, Shi; Furtaw, Michael D; Chen, Huaxian; Lamb, Don T; Ferguson, Stephen A; Arvin, Natalie E; Dawod, Mohamed; Kennedy, Robert T

2016-07-05

Western blotting is a commonly used protein assay that combines the selectivity of electrophoretic separation and immunoassay. The technique is limited by long time, manual operation with mediocre reproducibility, and large sample consumption, typically 10-20 μg per assay. Western blots are also usually used to measure only one protein per assay with an additional housekeeping protein for normalization. Measurement of multiple proteins is possible; however, it requires stripping membranes of antibody and then reprobing with a second antibody. Miniaturized alternatives to Western blot based on microfluidic or capillary electrophoresis have been developed that enable higher-throughput, automation, and greater mass sensitivity. In one approach, proteins are separated by electrophoresis on a microchip that is dragged along a polyvinylidene fluoride membrane so that as proteins exit the chip they are captured on the membrane for immunoassay. In this work, we improve this method to allow multiplexed protein detection. Multiple injections made from the same sample can be deposited in separate tracks so that each is probed with a different antibody. To further enhance multiplexing capability, the electrophoresis channel dimensions were optimized for resolution while keeping separation and blotting times to less than 8 min. Using a 15 μm deep × 50 μm wide × 8.6 cm long channel, it is possible to achieve baseline resolution of proteins that differ by 5% in molecular weight, e.g., ERK1 (44 kDa) from ERK2 (42 kDa). This resolution allows similar proteins detected by cross-reactive antibodies in a single track. We demonstrate detection of 11 proteins from 9 injections from a single Jurkat cell lysate sample consisting of 400 ng of total protein using this procedure. Thus, multiplexed Western blots are possible without cumbersome stripping and reprobing steps.

Allele-specific methylated multiplex real-time quantitative PCR (ASMM RTQ-PCR), a powerful method for diagnosing loss of imprinting of the 11p15 region in Russell Silver and Beckwith Wiedemann syndromes.

PubMed

Azzi, Salah; Steunou, Virginie; Rousseau, Alexandra; Rossignol, Sylvie; Thibaud, Nathalie; Danton, Fabienne; Le Jule, Marilyne; Gicquel, Christine; Le Bouc, Yves; Netchine, Irène

2011-02-01

Many human syndromes involve a loss of imprinting (LOI) due to a loss (LOM) or a gain of DNA methylation (GOM). Most LOI occur as mosaics and can therefore be difficult to detect with conventional methods. The human imprinted 11p15 region is crucial for the control of fetal growth, and LOI at this locus is associated with two clinical disorders with opposite phenotypes: Beckwith-Wiedemann syndrome (BWS), characterized by fetal overgrowth and a high risk of tumors, and Russell-Silver syndrome (RSS), characterized by intrauterine and postnatal growth restriction. Until recently, we have been using Southern blotting for the diagnosis of RSS and BWS. We describe here a powerful quantitative technique, allele-specific methylated multiplex real-time quantitative PCR (ASMM RTQ-PCR), for the diagnosis of these two complex disorders. We first checked the specificity of the probes and primers used for ASMM RTQ-PCR. We then carried out statistical validation for this method, on both retrospective and prospective populations of patients. This analysis demonstrated that ASMM RTQ-PCR is more sensitive than Southern blotting for detecting low degree of LOI. Moreover, ASMM RTQ-PCR is a very rapid, reliable, simple, safe, and cost effective method. © 2011 Wiley-Liss, Inc.

Serological investigation of enterococcal infections using western blot.

PubMed

Sulaiman, A; Rakita, R M; Arduino, R C; Patterson, J E; Steckelberg, J M; Singh, K V; Murray, B E

1996-10-01

To assess the usefulness of Western blot in the diagnosis of enterococcal infections, a pilot study was conducted with a newly developed Western blot using sera from patients with confirmed enterococcal infections. Sera from 17 of 19 patients with enterococcal endocarditis reacted strongly to enterococcal antigens on the Western blot, and most produced specific bands at molecular weights 98 kDa and 54 kDa. Sera from patients with bacteremic cholangitis and pyelonephritis reacted frequently as well, but the pattern of bands was different from that observed with endocarditis. Eighty-five percent of 26 sera tested from patients with bacteremia and associated deep-seated infections (endocarditis, cholangitis, and pyelonephritis) were positive on Western blot, compared to 30% of sera from bacteremic patients with no clinically determined deep focus of infection (p < 0.001).

Quantitative analysis of species specificity of two anti-parvalbumin antibodies for detecting southern hemisphere fish species demonstrating strong phylogenetic association.

PubMed

Liang, Ji; Tan, Chui Choo; Taylor, Steve L; Baumert, Joseph L; Lopata, Andreas L; Lee, N Alice

2017-12-15

This study aimed to develop a novel approach to determine the correlation between the parvalbumin (PAV) contents and their corresponding immunoreactivity (detectability) in southern hemisphere fish species. The immuno-detected PAV contents of the test fish species were estimated by a quantitative SDS-PAGE. A quantitative Enzyme-Linked ImmunoSorbent Assay (ELISA) was formatted to assess relative immunoreactivity of PAV. Sixteen species (forty-three percent) displayed a positive correlation with the anti-cod PAV polyclonal antibody, but no correlation with the anti-carp PAV monoclonal antibody. There was a strong phylogenetic association of the PAV immunoreactivity. Species from the order of Perciformes showed strong binding with both antibodies; whereas species from Salmoniformes, Ophidiiformes, Scombriformes, Scorpaeniformes, and Tetraodontiformes showed weak or no binding. This approach showed for the first time a statistical correlation between the PAV content and the immunoreactivity and allowed to rank the relative species/order specificity of the two antibodies for the southern hemisphere fish PAV. Copyright © 2017 Elsevier Ltd. All rights reserved.

A 15-year-long Southern blotting analysis of FMR1 to detect female carriers and for prenatal diagnosis of fragile X syndrome in Taiwan.

PubMed

Tzeng, C-C; Tsai, L-P; Chang, Y-K; Hung, Y-J; Chang, Y-Y; Su, Y-P; Jiang, J-J; Liang, H-M

2017-08-01

Here, we review the results of Southern blotting analyses of the FMR1 gene performed in our reference laboratory in Taiwan over a 15-year period. In total, 725 high-risk women with a family history of fragile X syndrome (FXS) or idiopathic intellectual disability, 3911 low-risk pregnant women without such family history, and prenatal diagnosis data for 32 foetuses from 24 carrier mothers were included. Only 2 carriers were in the low-risk group, which indicated a prevalence of 1 of 1955 women (95% confidence interval: 1/7156-1/539). A total of 100 carriers were found to be in the high-risk group, thus revealing a significantly higher frequency than the low-risk group (100/725 vs 2/3911, P<0.0001). Eight of the 14 foetuses that inherited the maternal mutant allele were verified to have a full mutation, with the smallest maternal pre-mutation allele carrying 56 CGG repeats. The overall findings confirmed that the carrier prevalence among low-risk women in Taiwan is significantly lower than that reported in western countries. Therefore, the most important step for preventing FXS in Taiwan would be to focus on high-risk women by promoting general awareness of this disease and spreading knowledge regarding the benefits of carrier screening and prenatal testing. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Other notable protein blotting methods: a brief review.

PubMed

Kurien, Biji T; Scofield, R Hal

2015-01-01

Proteins have been transferred from the gel to the membrane by a variety of methods. These include vacuum blotting, centrifuge blotting, electroblotting of proteins to Teflon tape and membranes for N- and C-terminal sequence analysis, multiple tissue blotting, a two-step transfer of low- and high-molecular-weight proteins, acid electroblotting onto activated glass, membrane-array method for the detection of human intestinal bacteria in fecal samples, protein microarray using a new black cellulose nitrate support, electrotransfer using square wave alternating voltage for enhanced protein recovery, polyethylene glycol-mediated significant enhancement of the immunoblotting transfer, parallel protein chemical processing before and during western blot and the molecular scanner concept, electronic western blot of matrix-assisted laser desorption/ionization mass spectrometric-identified polypeptides from parallel processed gel-separated proteins, semidry electroblotting of peptides and proteins from acid-urea polyacrylamide gels, transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes, and the display of K(+) channel proteins on a solid nitrocellulose support for assaying toxin binding. The quantification of proteins bound to PVDF membranes by elution of CBB, clarification of immunoblots on PVDF for transmission densitometry, gold coating of nonconductive membranes before matrix-assisted laser desorption/ionization tandem mass spectrometric analysis to prevent charging effect for analysis of peptides from PVDF membranes, and a simple method for coating native polysaccharides onto nitrocellulose are some of the methods involving either the manipulation of membranes with transferred proteins or just a passive transfer of antigens to membranes. All these methods are briefly reviewed in this chapter.

Western blotting using chemiluminescent substrates.

PubMed

Alegria-Schaffer, Alice

2014-01-01

Western blotting is a powerful and commonly used tool to identify and quantify a specific protein in a complex mixture (Towbin et al., 1979). The technique enables indirect detection of protein samples immobilized on a nitrocellulose or polyvinylidene fluoride (PVDF) membrane. Copyright © 2014 Elsevier Inc. All rights reserved.

Post-staining electroblotting for efficient and reliable peptide blotting.

PubMed

Lee, Der-Yen; Chang, Geen-Dong

2015-01-01

Post-staining electroblotting has been previously described to transfer Coomassie blue-stained proteins from polyacrylamide gel onto polyvinylidene difluoride (PVDF) membranes. Actually, stained peptides can also be efficiently and reliably transferred. Because of selective staining procedures for peptides and increased retention of stained peptides on the membrane, even peptides with molecular masses less than 2 kDa such as bacitracin and granuliberin R are transferred with satisfactory results. For comparison, post-staining electroblotting is about 16-fold more sensitive than the conventional electroblotting for visualization of insulin on the membrane. Therefore, the peptide blots become practicable and more accessible to further applications, e.g., blot overlay detection or immunoblotting analysis. In addition, the efficiency of peptide transfer is favorable for N-terminal sequence analysis. With this method, peptide blotting can be normalized for further analysis such as blot overlay assay, immunoblotting, and N-terminal sequencing for identification of peptide in crude or partially purified samples.

Genetic relatedness of orbiviruses by RNA-RNA blot hybridization

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

Bodkin, D.K.

1985-01-01

RNA-RNA blot hybridization was developed in order to identify type-specific genes among double-stranded (ds) RNA viruses, to assess the genetic relatedness of dsRNA viruses and to classify new strains. Viral dsRNA segments were electrophoresed through 10% polyacrylamide gels, transferred to membranes, and hybridized to (5'/sup 32/P)-pCp labeled genomic RNA from a related strain. Hybridization was performed at 52/sup 0/C, 50% formamide, 5X SSC. Under these conditions heterologous RNA species must share greater than or equal to 74% sequence homology in order to form stable dsRNA hybrids. Cognate genes of nine members of the Palyam serogroup of orbiviruses were identified andmore » their sequence relatedness to the prototype. Palyam virus, was determined. Reciprocal blot hybridizations were performed using radiolabeled genomic RNA of all members of the Palyam serogroup. Unique and variant genes were identified by lack of cross-homology or by weak homology between segments. Since genes 2 and 6 exhibited the highest degree of sequence variability, response to the vertebrate immune system may be a major cause of sequence divergence among members of a single serogroup. Changuinola serogroup isolates were compared by dot-blot hybridization, while Colorado tick fever (CTF) serogroup isolates were compared by the RNA-RNA blot hybridization procedure described for reovirus and Palyam serogroup isolates. Preliminary blot hybridization data were also obtained on the relatedness of members of different Orbivirus serogroups.« less

Skin blotting: a noninvasive technique for evaluating physiological skin status.

PubMed

Minematsu, Takeo; Horii, Motoko; Oe, Makoto; Sugama, Junko; Mugita, Yuko; Huang, Lijuan; Nakagami, Gojiro; Sanada, Hiromi

2014-06-01

The skin performs important structural and physiological functions, and skin assessment represents an important step in identifying skin problems. Although noninvasive techniques for assessing skin status exist, no such techniques for monitoring its physiological status are available. This study aimed to develop a novel skin-assessment technique known as skin blotting, based on the leakage of secreted proteins from inside the skin following overhydration in mice. The applicability of this technique was further investigated in a clinical setting. Skin blotting involves 2 steps: collecting proteins by attaching a damp nitrocellulose membrane to the surface of the skin, and immunostaining the collected proteins. The authors implanted fluorescein-conjugated dextran (F-DEX)-containing agarose gels into mice and detected the tissue distribution of F-DEX under different blotting conditions. They also analyzed the correlations between inflammatory cytokine secretion and leakage following ultraviolet irradiation in mice and in relation to body mass index in humans. The F-DEX in mice was distributed in the deeper and shallower layers of skin and leaked through the transfollicular and transepidermal routes, respectively. Ultraviolet irradiation induced tumor necrosis factor secretion in the epidermis in mice, which was detected by skin blotting, whereas follicular tumor necrosis factor was associated with body mass index in obese human subjects. These results support the applicability of skin blotting for skin assessment. Skin blotting represents a noninvasive technique for assessing skin physiology and has potential as a predictive and diagnostic tool for skin disorders.

A brief review of other notable protein blotting methods.

PubMed

Kurien, Biji T; Scofield, R Hal

2009-01-01

A plethora of methods have been used for transferring proteins from the gel to the membrane. These include centrifuge blotting, electroblotting of proteins to Teflon tape and membranes for N- and C-terminal sequence analysis, multiple tissue blotting, a two-step transfer of low and high molecular weight proteins, blotting of Coomassie Brilliant Blue (CBB)-stained proteins from polyacrylamide gels to transparencies, acid electroblotting onto activated glass, membrane-array method for the detection of human intestinal bacteria in fecal samples, protein microarray using a new black cellulose nitrate support, electrotransfer using square wave alternating voltage for enhanced protein recovery, polyethylene glycol-mediated significant enhancement of the immunoblotting transfer, parallel protein chemical processing before and during western blot and the molecular scanner concept, electronic western blot of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-identified polypeptides from parallel processed gel-separated proteins, semidry electroblotting of peptides and proteins from acid-urea polyacrylamide gels, transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes, and the display of K(+) channel proteins on a solid nitrocellulose support for assaying toxin binding. The quantification of proteins bound to PVDF membranes by elution of CBB, clarification of immunoblots on PVDF for transmission densitometry, gold coating of nonconductive membranes before MALDI tandem mass spectrometric analysis to prevent charging effect for analysis of peptides from PVDF membranes, and a simple method for coating native polysaccharides onto nitrocellulose are some of the methods involving either the manipulation of membranes with transferred proteins or just a passive transfer of antigens to membranes. All these methods are briefly reviewed in this chapter.

Analysis of immunostaining and western blotting of endothelin 1 and its receptors in mitral stenosis

PubMed Central

Leão, Sydney Correia; Dashwood, Michael R.; de Andrade, Mateus Santana; Santos, Nicolas Nascimento; Teles, Olivia Regina Lins Leal; de Souza, Williasmin Batista; Rodrigues, Tania Maria de Andrade

2015-01-01

Introduction Rheumatic Fever represents a serious public health problem in developing countries, with thousands of new cases each year. It is an autoimmune disease, which occurs in response to infection by streptococcus A. Objective The aim of this study was to evaluate the immunolabeling and protein expression for endothelin-1 and 3 (ET-1, ET-3) and its receptors (ETA, ETB) in rheumatic mitral valves. Methods Immunohistochemistry was used to identify ET-1/ET-3 and ETA/ETB receptors in rheumatic and control mitral valves. Quantitative analysis of immunostaining for ET-1/ET-3 and ETA/ETB receptors was performed. In addition, western blot analysis was carried out to assess protein levels in tissue samples. Results ET-1 and ETA receptor immunostaining predominated in stenotic valves, mainly associated with fibrotic regions, inflammatory areas and neovascularization. Quantitative analysis showed that the average area with positive expression of ET-1 was 18.21±14.96%. For ETA and ETB, the mean expressed areas were respectively 15.06±13.13% and 9.20±11.09%. ET-3 did not have a significant expression. The correlation between the expression of both endothelin receptors were strongly positive (R=0.74, P=0.02), but the correlation between ET-1 and its receptor were negative for both ETA (R=-0.37, P=0.25), and ETB (R=-0.14, P=0.39). This data was supported by western blot analysis. Conclusion The strong correlation between ET-1 and its receptors suggests that both play a role in the pathophysiology of rheumatic mitral valve stenosis and may potentially act as biomarkers of this disease. PMID:26107453

Failure analysis of blots for diesel engine intercooler

NASA Astrophysics Data System (ADS)

Ren, Ping; Li, Zongquan; Wu, Jiangfei; Guo, Yibin; Li, Wanyou

2017-05-01

In diesel generating sets, it will lead to the abominable working condition if the fault couldn’t be recovered when the bolt of intercooler cracks. This paper aims at the fault of the blots of diesel generator intercooler and completes the analysis of the static strength and fatigue strength. Static intensity is checked considering blot preload and thermal stress. In order to obtain the thermal stress of the blot, thermodynamic of intercooler is calculated according to the measured temperature. Based on the measured vibration response and the finite element model, using dynamic load identification technique, equivalent excitation force of unit was solved. In order to obtain the force of bolt, the excitation force is loaded into the finite element model. By considering the thermal stress and preload as the average stress while the mechanical stress as the wave stress, fatigue strength analysis has been accomplished. Procedure of diagnosis is proposed in this paper. Finally, according to the result of intensity verification the fatigue failure is validation. Thereby, further studies are necessary to verification the result of the intensity analysis and put forward some improvement suggestion.

A Proteomics Approach to the Protein Normalization Problem: Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting.

PubMed

Wiśniewski, Jacek R; Mann, Matthias

2016-07-01

Proteomics and other protein-based analysis methods such as Western blotting all face the challenge of discriminating changes in the levels of proteins of interest from inadvertent changes in the amount loaded for analysis. Mass-spectrometry-based proteomics can now estimate the relative and absolute amounts of thousands of proteins across diverse biological systems. We reasoned that this new technology could prove useful for selection of very stably expressed proteins that could serve as better loading controls than those traditionally employed. Large-scale proteomic analyses of SDS lysates of cultured cells and tissues revealed deglycase DJ-1 as the protein with the lowest variability in abundance among different cell types in human, mouse, and amphibian cells. The protein constitutes 0.069 ± 0.017% of total cellular protein and occurs at a specific concentration of 34.6 ± 8.7 pmol/mg of total protein. Since DJ-1 is ubiquitous and therefore easily detectable with several peptides, it can be helpful in normalization of proteomic data sets. In addition, DJ-1 appears to be an advantageous loading control for Western blot that is superior to those used commonly used, allowing comparisons between tissues and cells originating from evolutionarily distant vertebrate species. Notably, this is not possible by the detection and quantitation of housekeeping proteins, which are often used in the Western blot technique. The approach introduced here can be applied to select the most appropriate loading controls for MS-based proteomics or Western blotting in any biological system.

An alternative method for processing northern blots after capillary transfer.

PubMed

Nilsen, Timothy W

2015-03-02

Different laboratories use different methods for the prehybridization, hybridization, and washing steps of the northern blotting procedure. In this protocol, a northern blot is pretreated with Church and Gilbert hybridization buffer to block nonspecific probe-binding sites. The immobilized RNA is then hybridized to a DNA probe specific for the RNA of interest. Finally, the membrane is washed and subjected to autoradiography or phosphorimaging. The solutions and conditions described here may be ideal for those who prefer to use fewer ingredients in their solutions. This protocol is designed to achieve the same goals as other northern blotting approaches. It minimizes background (nonspecific adherence of probe to membrane and nonspecific hybridization) and maximizes specific hybridization to RNAs immobilized on a membrane. © 2015 Cold Spring Harbor Laboratory Press.

Measurement of hygromycin B phosphotransferase activity in crude mammalian cell extracts by a simple dot-blot assay.

PubMed

Sørensen, M S; Duch, M; Paludan, K; Jørgensen, P; Pedersen, F S

1992-03-15

Hygromycin B (Hy) resistance, encoded by the prokaryotic gene hph, is commonly used as a dominant selectable marker for gene transfer experiments in mammalian cells. We describe a simple, quantitative dot-blot assay for measuring the activity in crude mammalian cell extracts of Hy phosphotransferase, the product of the hph gene. The assay shows no cross interference with substrates for neomycin phosphotransferase II, the product of the commonly used marker gene neo; hph and neo may thus be useful as a set of two non-interfering selectable marker and reporter genes for gene transfer experiments in mammalian cells.

Prolonged incubation and stacked film exposure improve sensitivity in western blotting.

PubMed

Luo, Haitao; Rankin, Gary O; Straley, Shannon; Chen, Yi Charlie

2011-01-01

Western blotting is a basic technique for protein detection. For proteins of less abundance or antibodies of poorer quality, an increased sensitivity is often desired. Although it is commonly known that higher concentrations of antibodies and prolonged film exposure times will help improve sensitivity in western blots, both measures come with their own risks, and it is often unclear to which extent these measures should be applied. We conducted time-course studies to investigate protein-antibody interactions and primary antibody-secondary antibody interactions in western blotting. We also propose a protocol of stacked film exposure and have tested it in standard curves and cancer cell samples. Our study found that protein-primary antibody interactions and primary antibody-secondary antibody interactions could take a longer time than commonly used "one hour" or "overnight", and in some cases longer than 48h, to reach its maximum binding. We also show that the modified protocol of stacked film exposure works well for both standard curves and biological samples, reaching a maximum sensitivity in western blots without blurring target signals or increasing backgrounds. In addition to regular optimization of antibody concentrations and film exposure time, a prolonged incubation with antibodies and stacked film exposure will also help improve sensitivity and reduce background in western blotting. Copyright © 2011 Elsevier Inc. All rights reserved.

A fragile X male with a broad smear on southern blot analysis representing 100-500 CGG repeats and no methylation at the EagI site of the FMR-1 gene

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

Lachiewicz, A.M.; Spiridigliozzi, G.A.; McConkie-Rosell, A.

1996-08-09

Fragile X DNA studies were carried out on all obligate carriers of a large fragile X family with 10 mentally retarded individuals. One 64-year-old carrier man with an altered FMR-1 allele was not described as being mentally retarded or as having any limitations in function. He was married, raised 8 children, and worked as an auto mechanic. On examination, he had macrocephaly and mild macroorchidism but few of the other typical physical findings of males with fragile X syndrome. His Full Scale IQ is 73, and his Vineland Adaptive Behavior Composite is 73. On the Woodcock-Johnson Psycho-Educational Battery-Revised, he achievedmore » standard scores of 64 in Reading, 55 in Math, and 83 in Knowledge. His DNA findings showed a broad smear on Southern blot analysis of 100-500 CGG repeats and no methylation at the EagI site upstream of the FMR-1 protein coding region. His FMR-1 protein production is 12% of normal. His daughters all have large premutations, with somatic instability in the size of the CGG repeat lengths. They all have evidence of academic underachievement and 2 have physical characteristics frequently described in individuals with fragile X. 21 refs., 3 figs.« less

SDS-Polyacrylamide Electrophoresis and Western Blotting Applied to the Study of Asthma.

PubMed

García-Solaesa, Virginia; Abad, Sara Ciria

2016-01-01

Western blotting is used to analyze proteins after being separated by electrophoresis and subsequently electro-transferred to a membrane. Once immobilized, a specific protein can be identified through its reaction with a labeled antibody or antigen. It is a methodology commonly used in biomedical research such as asthma studies, to assess the pathways of inflammatory mediators involved in the disease.Here, we describe an example of western blotting to determine the factors involved in asthma. In this chapter, the methodology of western blotting is reviewed, paying attention on potential problems and giving interesting recommendations.

Quantitative Analysis of HIV-1 Preintegration Complexes

PubMed Central

Engelman, Alan; Oztop, Ilker; Vandegraaff, Nick; Raghavendra, Nidhanapati K.

2009-01-01

Retroviral replication proceeds through the formation of a provirus, an integrated DNA copy of the viral RNA genome. The linear cDNA product of reverse transcription is the integration substrate and two different integrase activities, 3′ processing and DNA strand transfer, are required for provirus formation. Integrase nicks the cDNA ends adjacent to phylogenetically-conserved CA dinucleotides during 3′ processing. After nuclear entry and locating a suitable chromatin acceptor site, integrase joins the recessed 3′-OHs to the 5′-phosphates of a double-stranded staggered cut in the DNA target. Integrase functions in the context of a large nucleoprotein complex, called the preintegration complex (PIC), and PICs are analyzed to determine levels of integrase 3′ processing and DNA strand transfer activities that occur during acute virus infection. Denatured cDNA end regions are monitored by indirect end-labeling to measure the extent of 3′ processing. Native PICs can efficiently integrate their viral cDNA into exogenously added target DNA in vitro, and Southern blotting or nested PCR assays are used to quantify the resultant DNA strand transfer activity. This study details HIV-1 infection, PIC extraction, partial purification, and quantitative analyses of integrase 3′ processing and DNA strand transfer activities. PMID:19233280

A Laboratory Exercise Illustrating the Sensitivity and Specificity of Western Blot Analysis

ERIC Educational Resources Information Center

Chang, Ming-Mei; Lovett, Janice

2011-01-01

Western blot analysis, commonly known as "Western blotting," is a standard tool in every laboratory where proteins are analyzed. It involves the separation of polypeptides in polyacrylamide gels followed by the electrophoretic transfer of the separated polypeptides onto a nitrocellulose or polyvinylidene fluoride membrane. A replica of the…

Detection of Y chromosome sequences in a 45,X/46,XXq - patient by Southern blot analysis of PCR-amplified DNA and fluorescent in situ hybridization (FISH)

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

Kocova, M.; Siegel, S.F.; Wenger, S.L.

1995-02-13

In some cases of gonadal dysgenesis, cytogenetic analysis seems to be discordant with the phenotype of the patients. We have applied techniques such as Southern blot analysis and fluorescent in situ hybridization (FISH) to resolve the phenotype/genotype discrepancy in a patient with ambiguous genitalia in whom the peripheral blood karotype was 45,X. Gonadectomy at age 7 months showed the gonadal tissue to be prepubertal testis on the left side and a streak gonad on the right. The karyotype obtained from the left gonad was 45,X/46,XXq- and that from the right gonad was 45,X. Three different techniques, PCR amplification, FISH, andmore » chromosome painting for X and Y chromosomes, confirmed the presence of Y chromosome sequences. Five different tissues were evaluated. The highest percentage of Y chromosome positive cells were detected in the left gonad, followed by the peripheral blood lymphocytes, skin fibroblasts, and buccal mucosa. No Y chromosomal material could be identified in the right gonad. Since the Xq- chromosome is present in the left gonad (testis), it is likely that the Xq- contains Y chromosomal material. Sophisticated analysis in this patient showed that she has at least 2 cell lines, one of which contains Y chromosomal material. These techniques elucidated the molecular basis of the genital ambiguity for this patient. When Y chromosome sequences are present in patients with Ullrich-Turner syndrome or gonadal dysgenesis, the risk for gonadal malignancy is significantly increased. Hence, molecular diagnostic methods to ascertain for the presence of Y chromosome sequences may expedite the evaluation of patients with the ambiguous genitalia. 21 refs., 4 figs., 2 tabs.« less

Use of a sensitive EnVision +-based detection system for Western blotting: avoidance of streptavidin binding to endogenous biotin and biotin-containing proteins in kidney and other tissues.

PubMed

Banks, Rosamonde E; Craven, Rachel A; Harnden, Patricia A; Selby, Peter J

2003-04-01

Western blotting remains a central technique in confirming identities of proteins, their quantitation and analysis of various isoforms. The biotin-avidin/streptavidin system is often used as an amplification step to increase sensitivity but in some tissues such as kidney, "nonspecific" interactions may be a problem due to high levels of endogenous biotin-containing proteins. The EnVision system, developed for immunohistochemical applications, relies on binding of a polymeric conjugate consisting of up to 100 peroxidase molecules and 20 secondary antibody molecules linked directly to an activated dextran backbone, to the primary antibody. This study demonstrates that it is also a viable and sensitive alternative detection system in Western blotting applications.

Effects of Reusing Gel Electrophoresis and Electrotransfer Buffers on Western Blotting.

PubMed

Heda, Ghanshyam D; Omotola, Oluwabukola B; Heda, Rajiv P; Avery, Jamie

2016-09-01

SDS-PAGE and Western blotting are 2 of the most commonly used biochemical methods for protein analysis. Proteins are electrophoretically separated based on their MWs by SDS-PAGE and then electrotransferred to a solid membrane surface for subsequent protein-specific analysis by immunoblotting, a procedure commonly known as Western blotting. Both of these procedures use a salt-based buffer, with the latter procedure consisting of methanol as an additive known for its toxicity. Previous reports present a contradictory view in favor or against reusing electrotransfer buffer, also known as Towbin's transfer buffer (TTB), with an aim to reduce the toxic waste. In this report, we present a detailed analysis of not only reusing TTB but also gel electrophoresis buffer (EB) on proteins of low to high MW range. Our results suggest that EB can be reused for at least 5 times without compromising the electrophoretic separation of mixture of proteins in an MW standard, BSA, and crude cell lysates. Additionally, reuse of EB did not affect the quality of subsequent Western blots. Successive reuse of TTB, on the other hand, diminished the signal of proteins of different MWs in a protein standard and a high MW membrane protein cystic fibrosis transmembrane-conductance regulator (CFTR) in Western blotting.

Treponema pallidum western blot: comparison with the FTA-ABS test as a confirmatory test for syphilis.

PubMed

Backhouse, J L; Nesteroff, S I

2001-01-01

We developed a Treponema pallidum Western blot and compared the results with Treponema pallidum particle agglutination (TPPA) and fluorescent treponemal antibody absorption (FTA-ABS) tests. The Western blot was deemed reactive if the serum reacted with at least three major antigenic bands (TpN47, TpN44.5, TpN17, TpN15). The sensitivities of the Western blot, TPPA and FTA-ABS, were all 100% and the specificities of the Western blot, TPPA and FTA-ABS were 100%, 100% and 94.5% respectively. In 52 problem sera, reactive in only one treponemal test, the agreement between the Western blot and TPPA (61.5%) was significantly better than between Western blot and FTA-ABS (38.5%). The individual sensitivities and specificities of TpN47, TpN44.5, TpN17, TpN15 were 100%, 100%, 96%, 100% and 20%, 96%, 100%, 100% respectively. We conclude that the Western blot is a useful additional confirmatory test or alternative to the FTA-ABS and that a more sensitive and specific criterion for the Western blot would be reactivity with TpN15 and two of the three other major antigens.

HIV‑2 antibody detection after indeterminate or negative HIV‑1 Western blot in Cuba, 2005-2008.

PubMed

Díaz, Dervel F; Ortiz, Eva; Martín, Dayamí; Nibot, Carmen; Rizo, Adis; Silva, Eladio

2012-01-01

INTRODUCTION Differentiating between HIV-1 and HIV-2 infection is the first step to understanding HIV transmission, epidemiology and pathogenesis in geographical areas where both viruses circulate. In Cuba, positive results in mixed HIV-1/2 screening assays are confirmed by HIV-1 Western blot. Indeterminate results constitute the main limitation of this test and HIV-2 infection is among their possible causes; hence the importance of second-stage screening and confirmatory tests for HIV-2 infection. OBJECTIVE Investigate the contribution of HIV-2 antibodies to negative or indeterminate HIV-1 Western blot results in serum samples from 2005 through 2008 in Cuba. METHODS HIV-2 reactivity was studied using the ELISA DAVIH-VIH-2 diagnostic kit (Cuba) in 1723 serum samples with negative or indeterminate results for HIV-1 Western blot from January 2005 through December 2008. Duplicate sera reactive by ELISA were confirmed by HIV-2 Western blot, results interpreted according to WHO criteria. The epidemiological interview established by Cuba's National Program for Prevention and Control Sexually-Transmitted Diseases and HIV/AIDS was applied to HIV-2 Western blot-positive patients. RESULTS Among all sera studied, HIV-2 ELISA identified 12 reactive serum samples (0.70%) and 1711 non-reactive (99.30%). Western blot analysis of the 12 ELISA-reactive samples confirmed two positive samples (16.67%), 4 negative (33.33%) and 6 indeterminate (50%). Positive samples reacted against the p16, p26, gp36, p53, p56, p68 and gp105 proteins. All 12 ELISA-reactive samples belonged to the HIV-1 Western blot indeterminate group. The two HIV-2-positive samples showed well defined reactivity to gp160, p53, p55 and p34 of HIV-1. HIV-1 seroconversion was observed in all 10 remaining samples during serological followup. CONCLUSIONS Two new HIV-2 seropositive cases were diagnosed using DAVIH-VIH-2 and HIV-2 Western blot in indeterminate HIV-1 Western blot samples. Results support the recommendation

An evaluation of new and established methods to determine T‐DNA copy number and homozygosity in transgenic plants.

PubMed Central

Głowacka, Katarzyna; Kromdijk, Johannes; Leonelli, Lauriebeth; Niyogi, Krishna K.; Clemente, Tom E.

2016-01-01

Abstract Stable transformation of plants is a powerful tool for hypothesis testing. A rapid and reliable evaluation method of the transgenic allele for copy number and homozygosity is vital in analysing these transformations. Here the suitability of Southern blot analysis, thermal asymmetric interlaced (TAIL‐)PCR, quantitative (q)PCR and digital droplet (dd)PCR to estimate T‐DNA copy number, locus complexity and homozygosity were compared in transgenic tobacco. Southern blot analysis and ddPCR on three generations of transgenic offspring with contrasting zygosity and copy number were entirely consistent, whereas TAIL‐PCR often underestimated copy number. qPCR deviated considerably from the Southern blot results and had lower precision and higher variability than ddPCR. Comparison of segregation analyses and ddPCR of T1 progeny from 26 T0 plants showed that at least 19% of the lines carried multiple T‐DNA insertions per locus, which can lead to unstable transgene expression. Segregation analyses failed to detect these multiple copies, presumably because of their close linkage. This shows the importance of routine T‐DNA copy number estimation. Based on our results, ddPCR is the most suitable method, because it is as reliable as Southern blot analysis yet much faster. A protocol for this application of ddPCR to large plant genomes is provided. PMID:26670088

Correlation of Leukocyte Telomere Length Measurement Methods in Patients with Dyskeratosis Congenita and in Their Unaffected Relatives.

PubMed

Khincha, Payal P; Dagnall, Casey L; Hicks, Belynda; Jones, Kristine; Aviv, Abraham; Kimura, Masayuki; Katki, Hormuzd; Aubert, Geraldine; Giri, Neelam; Alter, Blanche P; Savage, Sharon A; Gadalla, Shahinaz M

2017-08-13

Several methods have been employed to measure telomere length (TL) in human studies. It has been difficult to directly compare the results from these studies because of differences in the laboratory techniques and output parameters. We compared TL measurements (TLMs) by the three most commonly used methods, quantitative polymerase chain reaction (qPCR), flow cytometry with fluorescence in situ hybridization (flow FISH) and Southern blot, in a cohort of patients with the telomere biology disorder dyskeratosis congenita (DC) and in their unaffected relatives (controls). We observed a strong correlation between the Southern blot average TL and the flow FISH total lymphocyte TL in both the DC patients and their unaffected relatives ( R ² of 0.68 and 0.73, respectively). The correlation between the qPCR average TL and that of the Southern blot method was modest ( R ² of 0.54 in DC patients and of 0.43 in unaffected relatives). Similar results were noted when comparing the qPCR average TL and the flow FISH total lymphocyte TL ( R ² of 0.49 in DC patients and of 0.42 in unaffected relatives). In conclusion, the strengths of the correlations between the three widely used TL assays (qPCR, flow FISH, and Southern blot) were significantly different. Careful consideration is warranted when selecting the method of TL measurement for research and for clinical studies.

Lectin-Array Blotting.

PubMed

Pazos, Raquel; Echevarria, Juan; Hernandez, Alvaro; Reichardt, Niels-Christian

2017-09-01

Aberrant protein glycosylation is a hallmark of cancer, infectious diseases, and autoimmune or neurodegenerative disorders. Unlocking the potential of glycans as disease markers will require rapid and unbiased glycoproteomics methods for glycan biomarker discovery. The present method is a facile and rapid protocol for qualitative analysis of protein glycosylation in complex biological mixtures. While traditional lectin arrays only provide an average signal for the glycans in the mixture, which is usually dominated by the most abundant proteins, our method provides individual lectin binding profiles for all proteins separated in the gel electrophoresis step. Proteins do not have to be excised from the gel for subsequent analysis via the lectin array but are transferred by contact diffusion from the gel to a glass slide presenting multiple copies of printed lectin arrays. Fluorescently marked glycoproteins are trapped by the printed lectins via specific carbohydrate-lectin interactions and after a washing step their binding profile with up to 20 lectin probes is analyzed with a fluorescent scanner. The method produces the equivalent of 20 lectin blots in a single experiment, giving detailed insight into the binding epitopes present in the fractionated proteins. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

Line scanning system for direct digital chemiluminescence imaging of DNA sequencing blots

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

Karger, A.E.; Weiss, R.; Gesteland, R.F.

A cryogenically cooled charge-coupled device (CCD) camera equipped with an area CCD array is used in a line scanning system for low-light-level imaging of chemiluminescent DNA sequencing blots. Operating the CCD camera in time-delayed integration (TDI) mode results in continuous data acquisition independent of the length of the CCD array. Scanning is possible with a resolution of 1.4 line pairs/mm at the 50% level of the modulation transfer function. High-sensitivity, low-light-level scanning of chemiluminescent direct-transfer electrophoresis (DTE) DNA sequencing blots is shown. The detection of DNA fragments on the blot involves DNA-DNA hybridization with oligonucleotide-alkaline phosphatase conjugate and 1,2-dioxetane-based chemiluminescence.more » The width of the scan allows the recording of up to four sequencing reactions (16 lanes) on one scan. The scan speed of 52 cm/h used for the sequencing blots corresponds to a data acquisition rate of 384 pixels/s. The chemiluminescence detection limit on the scanned images is 3.9 [times] 10[sup [minus]18] mol of plasmid DNA. A conditional median filter is described to remove spikes caused by cosmic ray events from the CCD images. 39 refs., 9 refs.« less

PARP-1 serves as a novel molecular marker for hepatocellular carcinoma in a Southern Chinese Zhuang population.

PubMed

Li, Jiatong; Dou, Dongwei; Li, Ping; Luo, Wenqi; Lv, Wenxin; Zhang, Chengdong; Song, Xiaowei; Yang, Yuan; Zhang, Yuening; Xu, Yanzhen; Xiao, Feifan; Wei, Yan; Qin, Jian; Li, Hongtao; Yang, Xiaoli

2017-07-01

PARP-1 (poly(ADP-ribose) polymerase-1) plays an important role in tumorigenesis. Since its effects on different populations are varied, this study investigated the impact of PARP-1 on primary hepatocellular carcinoma in a Southern Chinese Zhuang population. We assessed the global PARP-1 messenger RNA expression in patients with hepatocellular carcinoma using The Cancer Genome Atlas dataset. Increased PARP-1 expression, related to alpha-fetoprotein level, was observed. The area under the receiver operating characteristic curve value was 0.833. Kaplan-Meier survival curves indicated that higher PARP-1 expression was not correlated with poorer overall survival and recurrence-free survival. In a Zhuang population, PARP-1 messenger RNA and protein levels were increased in the hepatocellular carcinoma tissue and its adjacent liver tissues as assessed by quantitative polymerase chain reaction, immunohistochemistry, and western blotting. Higher PARP-1 level was associated with a higher tumor stage (p < 0.05), without correlation with age, gender, smoking, drinking, tumor size, serum alpha-fetoprotein level, hepatitis B virus infection, metastasis, and invasion (p > 0.05). Further analysis suggested that H2AX, a PARP-1 protein interaction partner, was coordinated with PARP-1 in hepatocellular carcinoma tumorigenesis. Overall, some new characteristics of PARP-1 expression were noted in the Zhuang population. PARP-1 is a novel promising diagnostic marker for hepatocellular carcinoma in the Southern Chinese Zhuang population.

Lipid A binding proteins in macrophages detected by ligand blotting

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

Hampton, R.Y.; Golenbock, D.T.; Raetz, C.R.H.

1987-05-01

Endotoxin (LPS) stimulates a variety of eukaryotic cells. These actions are involved in the pathogenesis of Gram-negative septicemia. The site of action of the LPS toxic moiety, lipid A (LA), is unclear. Their laboratory has previously identified a bioactive LA precursor lipid IV/sub A/, which can be enzymatically labeled with /sup 32/P/sub i/ (10/sup 9/ dpm/nmole) and purified (99%). They now show that this ligand binds to specific proteins immobilized on nitrocellulose (NC) from LPS-sensitive RAW 264.7 cultured macrophages. NC blots were incubated with (/sup 32/P)-IV/sub A/ in a buffer containing BSA, NaCl, polyethylene glycol, and azide. Binding was assessedmore » using autoradiography or scintillation counting. Dot blot binding of the radioligand was inhibited by excess cold IV/sub A/, LA, or ReLPS but not by phosphatidylcholine, cardiolipin, phosphatidylinositol, or phosphatidic acid. Binding was trypsin-sensitive and dependent on protein concentration. Particulate macrophage proteins were subjected to SDS-PAGE and then electroblotted onto NC. Several discrete binding proteins were observed. Identical treatment of fetal bovine serum or molecular weight standards revealed no detectable binding. By avoiding high nonspecific binding of intact membranes, this ligand blotting assay may be useful in elucidating the molecular actions of LPS.« less

Quantitative assessment of the flow pattern in the southern Arava Valley (Israel) by environmental tracers and a mixing cell model

NASA Astrophysics Data System (ADS)

Adar, E. M.; Rosenthal, E.; Issar, A. S.; Batelaan, O.

1992-08-01

This paper demonstrates the implementation of a novel mathematical model to quantify subsurface inflows from various sources into the arid alluvial basin of the southern Arava Valley divided between Israel and Jordan. The model is based on spatial distribution of environmental tracers and is aimed for use on basins with complex hydrogeological structure and/or with scarce physical hydrologic information. However, a sufficient qualified number of wells and springs are required to allow water sampling for chemical and isotopic analyses. Environmental tracers are used in a multivariable cluster analysis to define potential sources of recharge, and also to delimit homogeneous mixing compartments within the modeled aquifer. Six mixing cells were identified based on 13 constituents. A quantitative assessment of 11 significant subsurface inflows was obtained. Results revealed that the total recharge into the southern Arava basin is around 12.52 × 10 6m3year-1. The major source of inflow into the alluvial aquifer is from the Nubian sandstone aquifer which comprises 65-75% of the total recharge. Only 19-24% of the recharge, but the most important source of fresh water, originates over the eastern Jordanian mountains and alluvial fans.

Avoiding pitfalls of internal controls: validation of reference genes for analysis by qRT-PCR and Western blot throughout rat retinal development.

PubMed

Rocha-Martins, Maurício; Njaine, Brian; Silveira, Mariana S

2012-01-01

Housekeeping genes have been commonly used as reference to normalize gene expression and protein content data because of its presumed constitutive expression. In this paper, we challenge the consensual idea that housekeeping genes are reliable controls for expression studies in the retina through the investigation of a panel of reference genes potentially suitable for analysis of different stages of retinal development. We applied statistical tools on combinations of retinal developmental stages to assess the most stable internal controls for quantitative RT-PCR (qRT-PCR). The stability of expression of seven putative reference genes (Actb, B2m, Gapdh, Hprt1, Mapk1, Ppia and Rn18s) was analyzed using geNorm, BestKeeper and Normfinder software. In addition, several housekeeping genes were tested as loading controls for Western blot in the same sample panel, using Image J. Overall, for qRT-PCR the combination of Gapdh and Mapk1 showed the highest stability for most experimental sets. Actb was downregulated in more mature stages, while Rn18s and Hprt1 showed the highest variability. We normalized the expression of cyclin D1 using various reference genes and demonstrated that spurious results may result from blind selection of internal controls. For Western blot significant variation could be seen among four putative internal controls (β-actin, cyclophilin b, α-tubulin and lamin A/C), while MAPK1 was stably expressed. Putative housekeeping genes exhibit significant variation in both mRNA and protein content during retinal development. Our results showed that distinct combinations of internal controls fit for each experimental set in the case of qRT-PCR and that MAPK1 is a reliable loading control for Western blot. The results indicate that biased study outcomes may follow the use of reference genes without prior validation for qRT-PCR and Western blot.

BLOTS AND ALL: A HISTORY OF THE RORSCHACH INK BLOT TEST IN BRITAIN.

PubMed

Hubbard, Katherine; Hegarty, Peter

2016-01-01

Despite the easily recognizable nature of the Rorschach ink blot test very little is known about the history of the test in Britain. We attend to the oft-ignored history of the Rorschach test in Britain and compare it to its history in the US. Prior to the Second World War, Rorschach testing in Britain had attracted advocates and critiques. Afterward, the British Rorschach Forum, a network with a high proportion of women, developed around the Tavistock Institute in London and The Rorschach Newsletter. In 1968, the International Rorschach Congress was held in London but soon after the group became less exclusive, and fell into decline. A comparative account of the Rorschach in Britain demonstrates how different national institutions invested in the 'projective hypothesis' according to the influence of psychoanalysis, the adoption of a nationalized health system, and the social positioning of 'others' throughout the twentieth century. In comparing and contrasting the history of the Rorschach in Britain and the US, we decentralize and particularize the history of North American Psychology. © 2016 Wiley Periodicals, Inc.

Evaluation of a new eastern blotting technique for the analysis of ginsenoside Re in American ginseng berry pulp extracts.

PubMed

Morinaga, Osamu; Uto, Takuhiro; Yuan, Chun-Su; Tanaka, Hiroyuki; Shoyama, Yukihiro

2010-06-01

A new eastern blotting technique has been established for ginsenoside Re (G-Re) contained in American ginseng berry pulp extracts. G-Re in American ginseng berry pulp was extracted using 100% methanol, 100% ethanol, 50% aqueous methanol, and 50% aqueous ethanol. The combined crude extracts were applied onto a polyethersulfone membrane and developed using the methanol-water-acetic acid solvent system (45:55:1 v/v). Separated components were immunostained using anti-G-Re monoclonal antibody. G-Re was first specifically detected and then quantitatively analyzed using NIH Imaging software. We also confirmed that the most suitable solvent was 50% aqueous methanol for extracting G-Re from American ginseng berry pulp. (c) 2009 Elsevier B.V. All rights reserved.

Protein analysis through Western blot of cells excised individually from human brain and muscle tissue

PubMed Central

Koob, A.O.; Bruns, L.; Prassler, C.; Masliah, E.; Klopstock, T.; Bender, A.

2016-01-01

Comparing protein levels from single cells in tissue has not been achieved through Western blot. Laser capture microdissection allows for the ability to excise single cells from sectioned tissue and compile an aggregate of cells in lysis buffer. In this study we analyzed proteins from cells excised individually from brain and muscle tissue through Western blot. After we excised individual neurons from the substantia nigra of the brain, the accumulated surface area of the individual cells was 120,000, 24,000, 360,000, 480,000, 600,000 μm2. We used an optimized Western blot protocol to probe for tyrosine hydroxylase in this cell pool. We also took 360,000 μm2 of astrocytes (1700 cells) and analyzed the specificity of the method. In muscle we were able to analyze the proteins of the five complexes of the electron transport chain through Western blot from 200 human cells. With this method, we demonstrate the ability to compare cell-specific protein levels in the brain and muscle and describe for the first time how to visualize proteins through Western blot from cells captured individually. PMID:22402104

Protein analysis through Western blot of cells excised individually from human brain and muscle tissue.

PubMed

Koob, A O; Bruns, L; Prassler, C; Masliah, E; Klopstock, T; Bender, A

2012-06-15

Comparing protein levels from single cells in tissue has not been achieved through Western blot. Laser capture microdissection allows for the ability to excise single cells from sectioned tissue and compile an aggregate of cells in lysis buffer. In this study we analyzed proteins from cells excised individually from brain and muscle tissue through Western blot. After we excised individual neurons from the substantia nigra of the brain, the accumulated surface area of the individual cells was 120,000, 24,000, 360,000, 480,000, 600,000 μm2. We used an optimized Western blot protocol to probe for tyrosine hydroxylase in this cell pool. We also took 360,000 μm2 of astrocytes (1700 cells) and analyzed the specificity of the method. In muscle we were able to analyze the proteins of the five complexes of the electron transport chain through Western blot from 200 human cells. With this method, we demonstrate the ability to compare cell-specific protein levels in the brain and muscle and describe for the first time how to visualize proteins through Western blot from cells captured individually. Copyright © 2012 Elsevier Inc. All rights reserved.

Blotting from PhastGel to Membranes by Ultrasound.

PubMed

Kost, Joseph; Azagury, Aharon

2015-01-01

Ultrasound based approach for enhanced protein blotting is proposed. Three minutes of ultrasound exposure (1 MHz, 2.5 W/cm(2)) was sufficient for a clear transfer of proteins from a polyacrylamide gel (PhastGel) to nitrocellulose or Nylon 66 Biotrans membrane. The proteins evaluated were prestained sodium dodecyl sulfate-polyacrylamide standards (18,500-106,000 Da) and 14C-labeled Rainbow protein molecular weight markers (14,300-200,000 Da).

Blotting from PhastGel to membranes by ultrasound.

PubMed

Kost, Joseph

2009-01-01

Ultrasound-based approach for enhanced protein blotting is proposed. Three minutes of ultrasound exposure (1 MHz, 2.5 W/cm(2)) was sufficient for a clear transfer of proteins from a polyacrylamide gel (PhastGel) to nitrocellulose or Nylon 66 Biotrans membrane. The proteins evaluated were prestained sodium dodecyl sulfate-polyacrylamide standards (18,500-106,000 Da) and (14)C-labeled Rainbow protein molecular weight markers (14,300-200,000 Da).

A quantitative and high-throughput assay of human papillomavirus DNA replication.

PubMed

Gagnon, David; Fradet-Turcotte, Amélie; Archambault, Jacques

2015-01-01

Replication of the human papillomavirus (HPV) double-stranded DNA genome is accomplished by the two viral proteins E1 and E2 in concert with host DNA replication factors. HPV DNA replication is an established model of eukaryotic DNA replication and a potential target for antiviral therapy. Assays to measure the transient replication of HPV DNA in transfected cells have been developed, which rely on a plasmid carrying the viral origin of DNA replication (ori) together with expression vectors for E1 and E2. Replication of the ori-plasmid is typically measured by Southern blotting or PCR analysis of newly replicated DNA (i.e., DpnI digested DNA) several days post-transfection. Although extremely valuable, these assays have been difficult to perform in a high-throughput and quantitative manner. Here, we describe a modified version of the transient DNA replication assay that circumvents these limitations by incorporating a firefly luciferase expression cassette in cis of the ori. Replication of this ori-plasmid by E1 and E2 results in increased levels of firefly luciferase activity that can be accurately quantified and normalized to those of Renilla luciferase expressed from a control plasmid, thus obviating the need for DNA extraction, digestion, and analysis. We provide a detailed protocol for performing the HPV type 31 DNA replication assay in a 96-well plate format suitable for small-molecule screening and EC50 determinations. The quantitative and high-throughput nature of the assay should greatly facilitate the study of HPV DNA replication and the identification of inhibitors thereof.

A quantitative image cytometry technique for time series or population analyses of signaling networks.

PubMed

Ozaki, Yu-ichi; Uda, Shinsuke; Saito, Takeshi H; Chung, Jaehoon; Kubota, Hiroyuki; Kuroda, Shinya

2010-04-01

Modeling of cellular functions on the basis of experimental observation is increasingly common in the field of cellular signaling. However, such modeling requires a large amount of quantitative data of signaling events with high spatio-temporal resolution. A novel technique which allows us to obtain such data is needed for systems biology of cellular signaling. We developed a fully automatable assay technique, termed quantitative image cytometry (QIC), which integrates a quantitative immunostaining technique and a high precision image-processing algorithm for cell identification. With the aid of an automated sample preparation system, this device can quantify protein expression, phosphorylation and localization with subcellular resolution at one-minute intervals. The signaling activities quantified by the assay system showed good correlation with, as well as comparable reproducibility to, western blot analysis. Taking advantage of the high spatio-temporal resolution, we investigated the signaling dynamics of the ERK pathway in PC12 cells. The QIC technique appears as a highly quantitative and versatile technique, which can be a convenient replacement for the most conventional techniques including western blot, flow cytometry and live cell imaging. Thus, the QIC technique can be a powerful tool for investigating the systems biology of cellular signaling.

Conditions that allow for effective transfer of membrane proteins onto nitrocellulose membrane in Western blots.

PubMed

Abeyrathne, Priyanka D; Lam, Joseph S

2007-04-01

A major hurdle in characterizing bacterial membrane proteins by Western blotting is the ineffectiveness of transferring these proteins from sodium dodecyl sulfate -- polyacrylamide gel electrophoresis (SDS-PAGE) gel onto nitrocellulose membrane, using standard Western blot buffers and electrophoretic conditions. In this study, we compared a number of modified Western blotting buffers and arrived at a composition designated as the SDS-PAGE-Urea Lysis buffer. The use of this buffer and specific conditions allowed the reproducible transfer of highly hydrophobic bacterial membrane proteins with 2-12 transmembrane-spanning segments as well as soluble proteins onto nitrocellulose membranes. This method should be broadly applicable for immunochemical studies of other membrane proteins.

Automated capillary Western dot blot method for the identity of a 15-valent pneumococcal conjugate vaccine.

PubMed

Hamm, Melissa; Ha, Sha; Rustandi, Richard R

2015-06-01

Simple Western is a new technology that allows for the separation, blotting, and detection of proteins similar to a traditional Western except in a capillary format. Traditionally, identity assays for biological products are performed using either an enzyme-linked immunosorbent assay (ELISA) or a manual dot blot Western. Both techniques are usually very tedious, labor-intensive, and complicated for multivalent vaccines, and they can be difficult to transfer to other laboratories. An advantage this capillary Western technique has over the traditional manual dot blot Western method is the speed and the automation of electrophoresis separation, blotting, and detection steps performed in 96 capillaries. This article describes details of the development of an automated identity assay for a 15-valent pneumococcal conjugate vaccine, PCV15-CRM197, using capillary Western technology. Copyright © 2015 Elsevier Inc. All rights reserved.

Human thyrotropin receptor subunits characterized by thyrotropin affinity purification and western blotting.

PubMed

Leedman, P J; Newman, J D; Harrison, L C

1989-07-01

We studied the subunit structure of the human TSH receptor in thyroid tissue from patients with Graves' disease and multinodular goiter by TSH affinity chromatography, immunoprecipitation with Graves' immunoglobulins (Igs), and a modified technique of Western blotting. Human TSH receptor-binding activity was purified about 1,270-fold by sequential affinity chromatography on wheat germ lectin-agarose and TSH-agarose. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of nonreduced affinity-purified receptors eluted in sodium dodecyl sulfate sample buffer revealed three noncovalently linked subunits of 70,000, 50,000, and 35,000 mol wt. When reduced, a major subunit of 25,000 mol wt was identified. When 3 mol/L NaCl was used to elute affinity-purified receptors only the 50,000 mol wt nonreduced subunit was detected. This subunit bound [125I]bovine TSH and was precipitated by Graves' Igs. Modifications to the conventional Western blotting technique enabled thyroglobulin components (approximately 220,000 mol wt), thyroid microsomal antigen (a doublet of approximately 110,000 mol wt), and putative TSH receptor subunits of 70,000 and 50,000 mol wt to be identified in thyroid particulate membranes by Graves' Igs. Blotting of affinity-purified receptors eluted in sodium dodecyl sulfate sample buffer revealed subunits of either 70,000 or 50,000 mol wt, with a minority of Graves' serum samples. We conclude that the nonreduced human TSH receptor is an oligomeric complex comprising three different subunits of 70,000, 50,000, and 35,000 mol wt. The reduced receptor exists as a single subunit of 25,000 mol wt, which may be disulfide linked to form the higher mol wt forms. The 70,000 and 50,000 mol wt subunits contain epitopes that bind Graves' Igs in modified Western blots, thus directly confirming that the human TSH receptor is a target for Graves' Igs.

Disseminated feline leishmaniosis due to Leishmania infantum in Southern France.

PubMed

Ozon, C; Marty, P; Pratlong, F; Breton, C; Blein, M; Lelièvre, A; Haas, P

1998-02-28

A fortuitously discovered case of feline leishmaniosis is reported. The parasites were found in the skin and the bone marrow of a domestic female cat that spontaneously died after a few weeks of evolution. Serological tests for FeLV, FIV and PIF virus detection gave negative results. By using Western blot serology, a characteristic pattern of leishmaniosis was obtained and by performing an isoenzyme electrophoresis, a Leishmania infantum MON-1 strain was identified. The same zymodeme is implicated in most of the canine and human leishmaniosis in Southern Europe. A study on the prevalence of asymptomatic feline leismaniosis is foreseen.

Quantitative estimate of commercial fish enhancement by seagrass habitat in southern Australia

NASA Astrophysics Data System (ADS)

Blandon, Abigayil; zu Ermgassen, Philine S. E.

2014-03-01

Seagrass provides many ecosystem services that are of considerable value to humans, including the provision of nursery habitat for commercial fish stock. Yet few studies have sought to quantify these benefits. As seagrass habitat continues to suffer a high rate of loss globally and with the growing emphasis on compensatory restoration, valuation of the ecosystem services associated with seagrass habitat is increasingly important. We undertook a meta-analysis of juvenile fish abundance at seagrass and control sites to derive a quantitative estimate of the enhancement of juvenile fish by seagrass habitats in southern Australia. Thirteen fish of commercial importance were identified as being recruitment enhanced in seagrass habitat, twelve of which were associated with sufficient life history data to allow for estimation of total biomass enhancement. We applied von Bertalanffy growth models and species-specific mortality rates to the determined values of juvenile enhancement to estimate the contribution of seagrass to commercial fish biomass. The identified species were enhanced in seagrass by 0.98 kg m-2 y-1, equivalent to ˜$A230,000 ha-1 y-1. These values represent the stock enhancement where all fish species are present, as opposed to realized catches. Having accounted for the time lag between fish recruiting to a seagrass site and entering the fishery and for a 3% annual discount rate, we find that seagrass restoration efforts costing $A10,000 ha-1 have a potential payback time of less than five years, and that restoration costing $A629,000 ha-1 can be justified on the basis of enhanced commercial fish recruitment where these twelve fish species are present.

β-actin as a loading control for plasma-based Western blot analysis of major depressive disorder patients.

PubMed

Zhang, Rufang; Yang, Deyu; Zhou, Chanjuan; Cheng, Ke; Liu, Zhao; Chen, Liang; Fang, Liang; Xie, Peng

2012-08-15

Western blot analysis is a commonly used technique for determining specific protein levels in clinical samples. For normalization of protein levels in Western blot, a suitable loading control is required. On account of its relatively high and constant expression, β-actin has been widely employed in Western blot of cell cultures and tissue extracts. However, β-actin's presence in human plasma and this protein's putative role as a plasma-based loading control for Western blot analysis remain unknown. In this study, an enzyme-linked immunosorbent assay was used to determine the concentration of β-actin in human plasma, which is 6.29±0.54 ng/ml. In addition, the linearity of β-actin immunostaining and loaded protein amount was evaluated by Western blot, and a fine linearity (R²=0.974±0.012) was observed. Furthermore, the expression of plasma β-actin in major depressive disorder subjects and healthy controls was compared. The data revealed no statistically significant difference between these two groups. Moreover, the total coefficient of variation for β-actin expression in the two groups was 9.2±1.2%. These findings demonstrate that β-actin is present in human plasma and may possibly be used as a suitable loading control for plasma-based Western blot analysis in major depressive disorder. Copyright © 2012 Elsevier Inc. All rights reserved.

Fluorescent Labeling of Proteins and Its Application to SDS-PAGE and Western Blotting.

PubMed

Alba, F Javier; Bartolomé, Salvador; Bermúdez, Antonio; Daban, Joan-Ramon

2015-01-01

This chapter describes very simple fluorescent methods developed in our laboratory allowing the rapid monitoring of total protein patterns on both sodium dodecyl sulfate (SDS) polyacrylamide gels and western blots. The noncovalent dye Nile red (9-diethylamino-5H-benzo[α]phenoxazine-5-one) is used for the sensitive staining of proteins in SDS gels. This method is compatible with the electroblotting of protein bands and with the staining of the resulting blot with the covalent dye MDPF (2-methoxy-2,4-diphenyl-3(2H)-furanone). These staining procedures are applied sequentially; there is no need to run a duplicate unstained gel for protein blotting. Furthermore, since only the adduct formed by the reaction of MDPF with proteins is fluorescent, there is no need to destain the membrane after protein labeling. In addition, MDPF staining is compatible with further immunodetection of specific bands with polyclonal antibodies. Finally, using the adequate conditions described below, MDPF staining does not preclude the N-terminal sequence analysis of proteins in selected bands.

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro.

PubMed

Jia, Zhaofeng; Liang, Yujie; Ma, Bin; Xu, Xiao; Xiong, Jianyi; Duan, Li; Wang, Daping

2017-05-17

The dedifferentiation of hyaline chondrocytes into fibroblastic chondrocytes often accompanies monolayer expansion of chondrocytes in vitro. The global DNA methylation level of chondrocytes is considered to be a suitable biomarker for the loss of the chondrocyte phenotype. However, results based on different experimental methods can be inconsistent. Therefore, it is important to establish a precise, simple, and rapid method to quantify global DNA methylation levels during chondrocyte dedifferentiation. Current genome-wide methylation analysis techniques largely rely on bisulfite genomic sequencing. Due to DNA degradation during bisulfite conversion, these methods typically require a large sample volume. Other methods used to quantify global DNA methylation levels include high-performance liquid chromatography (HPLC). However, HPLC requires complete digestion of genomic DNA. Additionally, the prohibitively high cost of HPLC instruments limits HPLC's wider application. In this study, genomic DNA (gDNA) was extracted from human chondrocytes cultured with varying number of passages. The gDNA methylation level was detected using a methylation-specific dot blot assay. In this dot blot approach, a gDNA mixture containing the methylated DNA to be detected was spotted directly onto an N + membrane as a dot inside a previously drawn circular template pattern. Compared with other gel electrophoresis-based blotting approaches and other complex blotting procedures, the dot blot method saves significant time. In addition, dot blots can detect overall DNA methylation level using a commercially available 5-mC antibody. We found that the DNA methylation level differed between the monolayer subcultures, and therefore could play a key role in chondrocyte dedifferentiation. The 5-mC dot blot is a reliable, simple, and rapid method to detect the general DNA methylation level to evaluate chondrocyte phenotype.

Semiquantitative Dot Blot with the GRA8 antigen to differentiate the stages of toxoplasmosis infection.

PubMed

Costa, Juan Gabriel; Vilariño, María Julia

2018-06-01

In this work we present a novel methodology to differentiate the phases of toxoplasmosis infection: the "semiquantitative Dot Blot". It is a simple technique that does not require expensive equipment, does not involve a long technique development, and can be used in a low-complexity laboratory. In this study, two recombinant sequences of Toxoplasma gondii GRA8 antigen were used, and specific IgG antibodies were detected in selected patient samples. This method makes it possible to obtain a score for each serum and define whether the patient is in the acute or chronic phase of the infection. The sensitivity and specificity results varied depending on the antigenic sequence used. With GRA8A, 62.1% and 72.7% were obtained, while with GRA8B, 82.8% and 72.1% were obtained, respectively. Although the sensitivity and specificity values were not close to 100%, they were similar to those reported with the same antigens in ELISA. Therefore, this quantitative technique would be a good alternative to ELISA. Copyright © 2018 Elsevier B.V. All rights reserved.

Quantum dot-based western blot for sensitive detection of pig serum antibody to actinobacillus pleuropneumoniae

NASA Astrophysics Data System (ADS)

Cişmileanu, Ana; Sima, Cornelia; Grigoriu, Constantin

2007-08-01

A quantum dot - immunoglobulin conjugate specific for pig IgG, was obtained by carbodiimide chemistry. We used a Western blot technique for detecting specific antibodies against Actinobacillus pleuropneumoniae (A. pp), which cause porcine pleuropneumonia. The antigen used in this technique was Apx haemolysin which is an important virulence factor of A. pp and it induces protective immunity in vaccined pigs. The detection on Western blot membrane was possible at 1/50 dilution of quantum dot conjugate at a dilution of pig serum till 1/6400. The results for pig serum demonstrated a higher sensitivity of QD-based Western blot technique for the presence of antibodies specific for Apx haemolysin in comparison with similar classical techniques (with coloured substrate for enzyme present in secondary antibody conjugate).

Label-Free Quantitative Proteomic Analysis of Chitosan Oligosaccharide-Treated Rice Infected with Southern Rice Black-Streaked Dwarf Virus.

PubMed

Yang, Anming; Yu, Lu; Chen, Zhuo; Zhang, Shanxue; Shi, Jing; Zhao, Xiaozhen; Yang, Yuanyou; Hu, Deyu; Song, Baoan

2017-05-18

Southern rice black-streaked dwarf virus (SRBSDV) has spread from thesouth of China to the north of Vietnam in the past few years and severelyinfluenced rice production. Its long incubation period and early symptoms are not evident; thus, controlling it is difficult. Chitosan oligosaccharide (COS) is a green plant immunomodulator. Early studies showed that preventing and controlling SRBSDV have a certain effect and reduce disease infection rate, but its underlying controlling and preventing mechanism is unclear. In this study, label-free proteomics was used to analyze differentially expressed proteins in rice after COS treatment. The results showed that COS can up-regulate the plant defense-related proteins and down-regulate the protein expression levels of SRBSDV. Meanwhile, quantitative real-time PCR test results showed that COS can improve defense gene expression in rice. Moreover, COS can enhance the defense enzymatic activities of peroxidase, superoxide dismutase and catalase through mitogen-activated protein kinase signaling cascade pathway, and enhance the rice disease resistance.

Evaluation of two commercial systems for automated processing, reading, and interpretation of Lyme borreliosis Western blots.

PubMed

Binnicker, M J; Jespersen, D J; Harring, J A; Rollins, L O; Bryant, S C; Beito, E M

2008-07-01

The diagnosis of Lyme borreliosis (LB) is commonly made by serologic testing with Western blot (WB) analysis serving as an important supplemental assay. Although specific, the interpretation of WBs for diagnosis of LB (i.e., Lyme WBs) is subjective, with considerable variability in results. In addition, the processing, reading, and interpretation of Lyme WBs are laborious and time-consuming procedures. With the need for rapid processing and more objective interpretation of Lyme WBs, we evaluated the performances of two automated interpretive systems, TrinBlot/BLOTrix (Trinity Biotech, Carlsbad, CA) and BeeBlot/ViraScan (Viramed Biotech AG, Munich, Germany), using 518 serum specimens submitted to our laboratory for Lyme WB analysis. The results of routine testing with visual interpretation were compared to those obtained by BLOTrix analysis of MarBlot immunoglobulin M (IgM) and IgG and by ViraScan analysis of ViraBlot and ViraStripe IgM and IgG assays. BLOTrix analysis demonstrated an agreement of 84.7% for IgM and 87.3% for IgG compared to visual reading and interpretation. ViraScan analysis of the ViraBlot assays demonstrated agreements of 85.7% for IgM and 94.2% for IgG, while ViraScan analysis of the ViraStripe IgM and IgG assays showed agreements of 87.1 and 93.1%, respectively. Testing by the automated systems yielded an average time savings of 64 min/run compared to processing, reading, and interpretation by our current procedure. Our findings demonstrated that automated processing and interpretive systems yield results comparable to those of visual interpretation, while reducing the subjectivity and time required for Lyme WB analysis.

Northern Blot Detection of Virus-Derived Small Interfering RNAs in Caenorhabditis elegans Using Nonradioactive Oligo Probes.

PubMed

Long, Tianyun; Lu, Rui

2017-01-01

Northern blot analysis has been widely used as a tool for detection and characterization of specific RNA molecules. When coupled with radioactive probe northern blot allows for robust detection and characterization of small RNA molecules of trace amount. Here, we describe the detection and size characterization of virus-derived small interfering RNAs (vsiRNAs) in C. elegans using nonradioactive DNA oligo probes in northern blotting. Our protocol allows for the detection and characterization of not only primary vsiRNAs but also secondary vsiRNAs, a class of single-stranded vsiRNAs that has distinct migration pattern, and can be easily adapted to the detection of vsiRNAs in other organisms.

Label-free DNA quantification via a 'pipette, aggregate and blot' (PAB) approach with magnetic silica particles on filter paper.

PubMed

Li, Jingyi; Liu, Qian; Alsamarri, Hussein; Lounsbury, Jenny A; Haversitick, Doris M; Landers, James P

2013-03-07

Reliable measurement of DNA concentration is essential for a broad range of applications in biology and molecular biology, and for many of these, quantifying the nucleic acid content is inextricably linked to obtaining optimal results. In its most simplistic form, quantitative analysis of nucleic acids can be accomplished by UV-Vis absorbance and, in more sophisticated format, by fluorimetry. A recently reported new concept, the 'pinwheel assay', involves a label-free approach for quantifying DNA through aggregation of paramagnetic beads in a rotating magnetic field. Here, we describe a simplified version of that assay adapted for execution using only a pipet and filter paper. The 'pipette, aggregate, and blot' (PAB) approach allows DNA to induce bead aggregation in a pipette tip through exposure to a magnetic field, followed by dispensing (blotting) onto filter paper. The filter paper immortalises the extent of aggregation, and digital images of the immortalized bead conformation, acquired with either a document scanner or a cell phone camera, allows for DNA quantification using a noncomplex algorithm. Human genomic DNA samples extracted from blood are quantified with the PAB approach and the results utilized to define the volume of sample used in a PCR reaction that is sensitive to input mass of template DNA. Integrating the PAB assay with paper-based DNA extraction and detection modalities has the potential to yield 'DNA quant-on-paper' devices that may be useful for point-of-care testing.

Southern Analysis of Genomic Alterations in Gamma-Ray-Induced Aprt- Hamster Cell Mutants

PubMed Central

Grosovsky, Andrew J.; Drobetsky, Elliot A.; deJong, Pieter J.; Glickman, Barry W.

1986-01-01

The role of genomic alterations in mutagenesis induced by ionizing radiation has been the subject of considerable speculation. By Southern blotting analysis we show here that 9 of 55 (approximately 1/6) gamma-ray-induced mutants at the adenine phosphoribosyl transferase (aprt) locus of Chinese hamster ovary (CHO) cells have a detectable genomic rearrangement. These fall into two classes: intragenic deletions and chromosomal rearrangements. In contrast, no major genomic alterations were detected among 67 spontaneous mutants, although two restriction site loss events were observed. Three gamma-ray-induced mutants were found to be intragenic deletions; all may have identical break-points. The remaining six gamma-ray-induced mutants demonstrating a genomic alteration appear to be the result of chromosomal rearrangements, possibly translocation or inversion events. None of the remaining gamma-ray-induced mutants showed any observable alteration in blotting pattern indicating a substantial role for point mutation in gamma-ray-induced mutagenesis at the aprt locus. PMID:3013724

A Study of Rubisco through Western Blotting and Tissue Printing Techniques

ERIC Educational Resources Information Center

Ma, Zhong; Cooper, Cynthia; Kim, Hyun-Joo; Janick-Buckner, Diane

2009-01-01

We describe a laboratory exercise developed for a cell biology course for second-year undergraduate biology majors. It was designed to introduce undergraduates to the basic molecular biology techniques of Western blotting and immunodetection coupled with the technique of tissue printing in detecting the presence, relative abundance, and…

Selection of a Clostridium perfringens type D epsilon toxin producer via dot-blot test.

PubMed

Gonçalves, Luciana A; Lobato, Zélia I P; Silva, Rodrigo O S; Salvarani, Felipe M; Pires, Prhiscylla S; Assis, Ronnie A; Lobato, Francisco C F

2009-11-01

Clostridium perfringens type D produces enterotoxemia, an enteric disease in ruminants, also known as pulpy kidney disease. Caused by epsilon toxin, enterotoxemia is a major exotoxin produced by this microorganism. Epsilon toxin is also the main component of vaccines against this enteric disorder. In this study, a standardized dot-blot was used to choose strains of C. perfringens type D that are producers of epsilon toxin. Clones producing epsilon toxin were chosen by limiting dilution; after three passages, lethal minimum dose titers were determined by soroneutralization test in mice. These clones produced epsilon toxin 240 times more concentrated than the original strain. The presence of the epsilon toxin gene (etx) was verified by polymerase chain reaction. All clones were positive, including those determined to be negative by dot-blot tests, suggesting that mechanisms in addition to the presence of the etx gene can influence toxin production. The dot-blot test was efficient for the selection of toxigenic colonies of C. perfringens type D and demonstrated that homogeneous populations selected from toxigenic cultures produce higher titers of epsilon toxin.

Comparison of recreational health risks associated with surfing and swimming in dry weather and post-storm conditions at Southern California beaches using quantitative microbial risk assessment (QMRA).

PubMed

Tseng, Linda Y; Jiang, Sunny C

2012-05-01

Southern California is an increasingly urbanized hotspot for surfing, thus it is of great interest to assess the human illness risks associated with this popular ocean recreational water sport from exposure to fecal bacteria contaminated coastal waters. Quantitative microbial risk assessments were applied to eight popular Southern California beaches using readily available enterococcus and fecal coliform data and dose-response models to compare health risks associated with surfing during dry weather and storm conditions. The results showed that the level of gastrointestinal illness risks from surfing post-storm events was elevated, with the probability of exceeding the US EPA health risk guideline up to 28% of the time. The surfing risk was also elevated in comparison with swimming at the same beach due to ingestion of greater volume of water. The study suggests that refinement of dose-response model, improving monitoring practice and better surfer behavior surveillance will improve the risk estimation. Copyright © 2012 Elsevier Ltd. All rights reserved.

Development of a nanogold slot blot inhibition assay for the detection of antibodies against bovine herpesvirus type 1.

PubMed

Japolla, Greice; Cunha-Junior, Jair Pereira; Pajuaba, Ana Claudia Arantes Marquez; Taketomi, Ernesto Akio; Bührer-Sékula, Samira; Bataus, Luiz Artur Mendes; de Souza, Guilherme Rocha Lino

2018-06-01

Bovine herpesvirus type 1 (BoHV-1) is recognized as an important pathogen causing respiratory, reproductive, and neurological disorders in cattle and is associated with economic losses to animal industry. Accurate diagnostic methods are needed for prevention of disease transmission. While the virus neutralization test is considered the gold standard method, it requires maintenance of the virus and cell cultures, which is time consuming and expensive. Serological techniques such as enzyme-linked immunosorbent assay (ELISA) are widely applied, as these are easy to perform and provide quick results. In the present study, a nanogold slot blot inhibition assay was developed for the serological diagnosis of BoHV-1 and compared with standard ELISA and horseradish peroxidase (HRP) slot blot assays. Of 42 serum samples tested by ELISA, 32 (76.2%) were positive and 10 (23.8%), were negative. The sensitivity and specificity of the nanogold slot blot inhibition assay was similar to that observed for ELISA and HRP slot blot assays, and a strong correlation was observed between the tests. Thus, the nanogold slot blot inhibition assay may serve as an efficient and rapid alternative to ELISA in settings, where plate-reading equipment is lacking.

Improved high-throughput quantification of luminescent microplate assays using a common Western-blot imaging system.

PubMed

Hawkins, Liam J; Storey, Kenneth B

2017-01-01

Common Western-blot imaging systems have previously been adapted to measure signals from luminescent microplate assays. This can be a cost saving measure as Western-blot imaging systems are common laboratory equipment and could substitute a dedicated luminometer if one is not otherwise available. One previously unrecognized limitation is that the signals captured by the cameras in these systems are not equal for all wells. Signals are dependent on the angle of incidence to the camera, and thus the location of the well on the microplate. Here we show that: •The position of a well on a microplate significantly affects the signal captured by a common Western-blot imaging system from a luminescent assay.•The effect of well position can easily be corrected for.•This method can be applied to commercially available luminescent assays, allowing for high-throughput quantification of a wide range of biological processes and biochemical reactions.

A rapid chemiluminescent slot blot immunoassay for the detection and quantification of Clostridium botulinum neurotoxin type E, in cultures.

PubMed

Cadieux, Brigitte; Blanchfield, Burke; Smith, James P; Austin, John W

2005-05-01

A simple, rapid, cost-effective in vitro slot blot immunoassay was developed for the detection and quantification of botulinum neurotoxin type E (BoNT/E) in cultures. Culture supernatants of 36 strains of clostridia, including 12 strains of Clostridium botulinum type E, 12 strains of other C. botulinum neurotoxin serotypes, and 12 strains of other clostridial species were tested. Samples containing BoNT/E were detected using affinity-purified polyclonal rabbit antisera prepared against BoNT/E with subsequent detection of secondary antibodies using chemiluminescence. All strains of C. botulinum type E tested positive, while all non C. botulinum type E strains tested negative. The sensitivity of the slot blot immunoassay for detection of BoNT/E was approximately four mouse lethal doses (MLD). The intensity of chemiluminescence was directly correlated with the concentration of BoNT/E up to 128 MLD, allowing quantification of BoNT/E between 4 and 128 MLD. The slot blot immunoassay was compared to the mouse bioassay for detection of BoNT/E using cultures derived from fish samples inoculated with C. botulinum type E, and cultures derived from naturally contaminated environmental samples. A total of 120 primary enrichment cultures derived from fish samples, of which 103 were inoculated with C. botulinum type E, and 17 were uninoculated controls, were assayed. Of the 103 primary enrichment cultures derived from inoculated fish samples, all were positive by mouse bioassay, while 94 were also positive by slot blot immunoassay, resulting in a 7.5% false-negative rate. All 17 primary enrichment cultures derived from the uninoculated fish samples were negative by both mouse bioassay and slot blot immunoassay. A total of twenty-six primary enrichment cultures derived from environmental samples were tested by mouse bioassay and slot blot immunoassay. Of 13 primary enrichment cultures positive by mouse bioassay, 12 were also positive by slot blot immunoassay, resulting in a 3

Ferulic acid enhances IgE binding to peanut allergens in western blots.

USDA-ARS?s Scientific Manuscript database

Phenolic compounds at high concentrations are known to form insoluble complexes with proteins. We hypothesized that this complex formation could interfere with Western blot and ELISA assays for peanut allergens. To verify this, three simple phenolic compounds (ferulic, caffeic, and chlorogenic acids...

COMPARISONS OF ELISA AND WESTERN BLOT ASSAYS FOR DETECTION OF CRYPTOSPORIDIUM ANTIBODY

EPA Science Inventory

A seroprevalence survey was conducted using ELISA and Western blot (WB) assays for antibody to three Cryptosporidium antigens on 380 blood donors in Jackson County, Oregon. The purpose was to determine if either assay could detect serological evidence of an outbreak which occurre...

Ferulic acid enhances IgE binding to peanut allergens in western blots.

USDA-ARS?s Scientific Manuscript database

Because phenolic compounds can precipitate or complex with proteins, we postulated that interactions of phenolics with IgE antibodies help enhance IgE binding to peanut allergens in Western blots. Three different phenolics, such as, ferulic, caffeic and chlorogenic acids were examined. Each was mixe...

Serological differentiation of murine typhus and epidemic typhus using cross-adsorption and Western blotting.

PubMed

La Scola, B; Rydkina, L; Ndihokubwayo, J B; Vene, S; Raoult, D

2000-07-01

Differentiation of murine typhus due to Rickettsia typhi and epidemic typhus due to Rickettsia prowazekii is critical epidemiologically but difficult serologically. Using serological, epidemiological, and clinical criteria, we selected sera from 264 patients with epidemic typhus and from 44 patients with murine typhus among the 29,188 tested sera in our bank. These sera cross-reacted extensively in indirect fluorescent antibody assays (IFAs) against R. typhi and R. prowazekii, as 42% of the sera from patients with epidemic typhus and 34% of the sera from patients with murine typhus exhibited immunoglobulin M (IgM) and/or IgG titers against the homologous antigen (R. prowazekii and R. typhi, respectively) that were more than one dilution higher than those against the heterologous antigen. Serum cross-adsorption studies and Western blotting were performed on sera from 12 selected patients, 5 with murine typhus, 5 with epidemic typhus, and 2 suffering from typhus of undetermined etiology. Differences in IFA titers against R. typhi and R. prowazekii allowed the identification of the etiological agent in 8 of 12 patients. Western blot studies enabled the identification of the etiological agent in six patients. When the results of IFA and Western blot studies were considered in combination, identification of the etiological agent was possible for 10 of 12 patients. Serum cross-adsorption studies enabled the differentiation of the etiological agent in all patients. Our study indicates that when used together, Western blotting and IFA are useful serological tools to differentiate between R. prowazekii and R. typhi exposures. While a cross-adsorption study is the definitive technique to differentiate between infections with these agents, it was necessary in only 2 of 12 cases (16.7%), and the high costs of such a study limit its use.

Serological Differentiation of Murine Typhus and Epidemic Typhus Using Cross-Adsorption and Western Blotting

PubMed Central

La Scola, Bernard; Rydkina, Lena; Ndihokubwayo, Jean-Bosco; Vene, Sirkka; Raoult, Didier

2000-01-01

Differentiation of murine typhus due to Rickettsia typhi and epidemic typhus due to Rickettsia prowazekii is critical epidemiologically but difficult serologically. Using serological, epidemiological, and clinical criteria, we selected sera from 264 patients with epidemic typhus and from 44 patients with murine typhus among the 29,188 tested sera in our bank. These sera cross-reacted extensively in indirect fluorescent antibody assays (IFAs) against R. typhi and R. prowazekii, as 42% of the sera from patients with epidemic typhus and 34% of the sera from patients with murine typhus exhibited immunoglobulin M (IgM) and/or IgG titers against the homologous antigen (R. prowazekii and R. typhi, respectively) that were more than one dilution higher than those against the heterologous antigen. Serum cross-adsorption studies and Western blotting were performed on sera from 12 selected patients, 5 with murine typhus, 5 with epidemic typhus, and 2 suffering from typhus of undetermined etiology. Differences in IFA titers against R. typhi and R. prowazekii allowed the identification of the etiological agent in 8 of 12 patients. Western blot studies enabled the identification of the etiological agent in six patients. When the results of IFA and Western blot studies were considered in combination, identification of the etiological agent was possible for 10 of 12 patients. Serum cross-adsorption studies enabled the differentiation of the etiological agent in all patients. Our study indicates that when used together, Western blotting and IFA are useful serological tools to differentiate between R. prowazekii and R. typhi exposures. While a cross-adsorption study is the definitive technique to differentiate between infections with these agents, it was necessary in only 2 of 12 cases (16.7%), and the high costs of such a study limit its use. PMID:10882661

Quantitative proteomic analysis for high-throughput screening of differential glycoproteins in hepatocellular carcinoma serum

PubMed Central

Gao, Hua-Jun; Chen, Ya-Jing; Zuo, Duo; Xiao, Ming-Ming; Li, Ying; Guo, Hua; Zhang, Ning; Chen, Rui-Bing

2015-01-01

Objective Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths. Novel serum biomarkers are required to increase the sensitivity and specificity of serum screening for early HCC diagnosis. This study employed a quantitative proteomic strategy to analyze the differential expression of serum glycoproteins between HCC and normal control serum samples. Methods Lectin affinity chromatography (LAC) was used to enrich glycoproteins from the serum samples. Quantitative mass spectrometric analysis combined with stable isotope dimethyl labeling and 2D liquid chromatography (LC) separations were performed to examine the differential levels of the detected proteins between HCC and control serum samples. Western blot was used to analyze the differential expression levels of the three serum proteins. Results A total of 2,280 protein groups were identified in the serum samples from HCC patients by using the 2D LC-MS/MS method. Up to 36 proteins were up-regulated in the HCC serum, whereas 19 proteins were down-regulated. Three differential glycoproteins, namely, fibrinogen gamma chain (FGG), FOS-like antigen 2 (FOSL2), and α-1,6-mannosylglycoprotein 6-β-N-acetylglucosaminyltransferase B (MGAT5B) were validated by Western blot. All these three proteins were up-regulated in the HCC serum samples. Conclusion A quantitative glycoproteomic method was established and proven useful to determine potential novel biomarkers for HCC. PMID:26487969

Wheat-specific gene, ribosomal protein l21, used as the endogenous reference gene for qualitative and real-time quantitative polymerase chain reaction detection of transgenes.

PubMed

Liu, Yi-Ke; Li, He-Ping; Huang, Tao; Cheng, Wei; Gao, Chun-Sheng; Zuo, Dong-Yun; Zhao, Zheng-Xi; Liao, Yu-Cai

2014-10-29

Wheat-specific ribosomal protein L21 (RPL21) is an endogenous reference gene suitable for genetically modified (GM) wheat identification. This taxon-specific RPL21 sequence displayed high homogeneity in different wheat varieties. Southern blots revealed 1 or 3 copies, and sequence analyses showed one amplicon in common wheat. Combined analyses with sequences from common wheat (AABBDD) and three diploid ancestral species, Triticum urartu (AA), Aegilops speltoides (BB), and Aegilops tauschii (DD), demonstrated the presence of this amplicon in the AA genome. Using conventional qualitative polymerase chain reaction (PCR), the limit of detection was 2 copies of wheat haploid genome per reaction. In the quantitative real-time PCR assay, limits of detection and quantification were about 2 and 8 haploid genome copies, respectively, the latter of which is 2.5-4-fold lower than other reported wheat endogenous reference genes. Construct-specific PCR assays were developed using RPL21 as an endogenous reference gene, and as little as 0.5% of GM wheat contents containing Arabidopsis NPR1 were properly quantified.

Proceedings of the 15th biennial southern silvicultural research conference

Treesearch

James M. Guldin

2013-01-01

Sixty-eight papers and seventeen posters address a range of issues affecting southern forests. Papers are grouped in 12 sessions that include pine silviculture session I, hardwood silviculture - intermediate treatment and stand development, longleaf pine; quantitative silviculture and economics, pine silviculture session II, hardwood regeneration, carbon and bioenergy...

Checking transfer efficiency and equal loading via qualitative optical way in western blotting.

PubMed

Gong, Jun-Hua; Gong, Jian-Ping; Zheng, Kai-Wen

2017-11-01

The ability to determine that successful transfer and equal loading occur prior to using primary antibodies is important. And total protein staining is commonly used to check transfer efficiency and normalization, which play a crucial role in western blotting. Ponceau S and coomassie blue are commonly used, but there are disadvantages reported in recent years. Therefore, we are interested in finding another method, which is cheap, easy and fast. As we know, protein binding region of PVDF membrane is still hydrophilic when carbinol volatilizes, however, the non-protein binding region of PVDF membrane became hydrophobic again. And this different wettability between non-protein binding region and protein binding region of Polyvinylidene difluoride membrane may be used to check transfer efficiency and equal loading in western blotting. Based on the principle above, we describe an optical approach where an experimenter can observe that the proteins have been transferred to the membrane without any staining within minutes. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Comparison of satellite-derived dynamical quantities for the stratosphere of the Southern Hemisphere

NASA Technical Reports Server (NTRS)

Miles, Thomas (Editor); Oneill, Alan (Editor)

1989-01-01

As part of the international Middle Atmosphere Program (MAP), a project was instituted to study the dynamics of the Middle Atmosphere in the Southern Hemisphere (MASH). A pre-MASH workshop was held with two aims: comparison of Southern Hemisphere dynamical quantities derived from various archives of satellite data; and assessing the impact of different base-level height information on such derived quantities. The dynamical quantities examined included geopotential height, zonal wind, potential vorticity, eddy heat and momentum fluxes, and Eliassen-Palm fluxes. It was found that while there was usually qualitative agreement between the different sets of fields, substantial quantitative differences were evident, particularly in high latitudes. The fidelity of the base-level analysis was found to be of prime importance in calculating derived quantities - especially the Eliassen-Palm flux divergence and potential vorticity. Improvements in base-level analyses are recommended. In particular, quality controls should be introduced to remove spurious localized features from analyses, and information from all Antarctic radiosondes should be utilized where possible. Caution in drawing quantitative inferences from satellite data for the middle atmosphere of the Southern Hemisphere is advised.

Quantitative assessment of hTERT mRNA expression in dysplastic nodules of HBV-related hepatocarcinogenesis.

PubMed

Oh, Bong-Kyeong; Kim, Young-Joo; Park, Young Nyun; Choi, Jinsub; Kim, Kyung Sik; Park, Chanil

2006-04-01

Telomerase reverse transcriptase (hTERT) is the rate-limiting determinant of telomerase, which is critical for carcinogenesis. Dysplastic nodules (DNs) appear to be preneoplastic lesions of hepatocellular carcinomas (HCCs). In this study, in order to characterize DNs, hTERT mRNA, hTERT gene dosage, and mRNA for c-myc, a transcriptional activator of hTERT were studied in human multi-step hepatocarcinogenesis. Fifty four hepatic nodules including 5 large regenerative nodules, 14 low-grade DNs, 7 high-grade DNs, 11 DNs with HCC foci and 17 HCCs, 23 livers with chronic hepatitis/cirrhosis, and 6 normal livers were examined. Transcript levels were measured by real-time quantitative RT-PCR and gene dosages by real-time PCR and Southern blotting. The hTERT mRNA levels increased with the progression of hepatocarcinogenesis, and a significant induction in the transition between low- and high-grade DNs was seen. Most high-grade DNs strongly expressed hTERT mRNA at levels similar to those of HCCs. Twenty-one percent of low-grade DNs had high levels of hTERT mRNA, up to those of high-grade DNs and there was no difference in the pathological features between low-grade DNs with and without increased hTERT mRNA levels. No correlation was found between hTERT mRNA levels, hTERT gene dosage, and c-myc mRNA levels. These results suggest that the induction of hTERT mRNA is an important early event and that its measurement by real-time quantitative RT-PCR is a useful tool to detect premalignant/malignant tendencies in hepatic nodules. However, hTERT gene dosage and c-myc expression are not the main mechanisms regulating hTERT expression in hepatocarcinogenesis.

Unraveling Molecular Differences of Gastric Cancer by Label-Free Quantitative Proteomics Analysis.

PubMed

Dai, Peng; Wang, Qin; Wang, Weihua; Jing, Ruirui; Wang, Wei; Wang, Fengqin; Azadzoi, Kazem M; Yang, Jing-Hua; Yan, Zhen

2016-01-21

Gastric cancer (GC) has significant morbidity and mortality worldwide and especially in China. Its molecular pathogenesis has not been thoroughly elaborated. The acknowledged biomarkers for diagnosis, prognosis, recurrence monitoring and treatment are lacking. Proteins from matched pairs of human GC and adjacent tissues were analyzed by a coupled label-free Mass Spectrometry (MS) approach, followed by functional annotation with software analysis. Nano-LC-MS/MS, quantitative real-time polymerase chain reaction (qRT-PCR), western blot and immunohistochemistry were used to validate dysregulated proteins. One hundred forty-six dysregulated proteins with more than twofold expressions were quantified, 22 of which were first reported to be relevant with GC. Most of them were involved in cancers and gastrointestinal disease. The expression of a panel of four upregulated nucleic acid binding proteins, heterogeneous nuclear ribonucleoprotein hnRNPA2B1, hnRNPD, hnRNPL and Y-box binding protein 1 (YBX-1) were validated by Nano-LC-MS/MS, qRT-PCR, western blot and immunohistochemistry assays in ten GC patients' tissues. They were located in the keynotes of a predicted interaction network and might play important roles in abnormal cell growth. The label-free quantitative proteomic approach provides a deeper understanding and novel insight into GC-related molecular changes and possible mechanisms. It also provides some potential biomarkers for clinical diagnosis.

Glycosaminoglycan blotting on nitrocellulose membranes treated with cetylpyridinium chloride after agarose-gel electrophoretic separation.

PubMed

Maccari, Francesca; Volpi, Nicola

2002-09-01

We describe a method for blotting and immobilizing several nonsulfated and sulfated complex polysaccharides on membranes made hydrophilic and positively charged by a cationic detergent after their separation by conventional agarose gel electrophoresis. Nitrocellulose membranes were derivatized with the cationic detergent cetylpyridinium chloride (CPC) and mixtures of glycosaminoglycans (GAGs) were capillary-blotted after their separation in agarose gel electrophoresis in barium acetate/1,2-diaminopropane. Single purified species of variously sulfated polysaccharides were transferred onto the derivatized membranes after electrophoresis with an efficiency of 100% and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining) permitting about 0.1 nug threshold of detection. Nonsulfated polyanions, hyaluronic acid, a fructose-containing polysaccharide with a chondroitin backbone purified from Escherichia coli U1-41, and its defructosylated product, were also electrophoretically separated and transferred onto membranes. The limit of detection for desulfated GAGs was about 0.1-0.5 nug after irreversible or reversible staining. GAG extracts from bovine, lung and aorta, and human aorta and urine were separated by agarose gel electrophoresis and blotted on CPC-treated nitrocellulose membranes. The polysaccharide composition of these extracts was determined. The membrane stained with toluidine blue (reversible staining) was destained and the same lanes used for immunological detection or other applications. Reversible staining was also applied to recover single species of polysaccharides after electrophoretic separation of mixtures of GAGs and their transfer onto membranes. Single bands were released from the membrane with an efficiency of 70-100% for further biochemical characterization.

Development of a Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS) Method for the Quantitation of Viral Envelope Glycoprotein in Ebola Virus-Like Particle Vaccine Preparations

DTIC Science & Technology

2016-09-05

46 was performed on an LTQ-Orbitrap Elite MS and the final quantitation was derived by 47 comparing the relative response of the 200 fmol AQUA...shown in Figure 3B, the final quantitation is derived by comparing the 527 relative response of the 200 fmol AQUA standards (SEE and IRSEE: Set 1) to...measure of eVLP quality, the western blot 553 and LC-HRMS quantitation results were compared to survival data in mice for each of these 554 eVLP vaccine

Comparative evaluation of western blotting in hepatic and pulmonary cystic echinococcosis.

PubMed

Akisu, C; Delibas, S B; Bicmen, C; Ozkoc, S; Aksoy, U; Turgay, N

2006-12-01

Many serological tests are widely used in the diagnosis of cystic echinococcosis (CE), caused by the larval stages of Echinococcus granulosus. The present study was carried for differentiation between hepatic and pulmonary cystic echinococcosis by Western Blotting (WB). A total of 121 sera from patients with hepatic CE (37), pulmonary CE (31) and controls (53; consisting of six healthy, seven Hymenolepis nana infection, 20 hepatic and 20 pulmonary diseases other than CE) were examined. In all of the CE patients, E. gronulosus infection was confirmed by surgical intervention. Sera were previously tested using IHA and ELISA to detect the E. gronulosus specific antibodies. Sera from hepatic cases of CE reacted with 16 polypeptides of 6-116 kDa and sera from pulmonary cases of CE reacted with 14 polypeptides of 4-130 kDa by Western Blotting. The WB test enabled the detection of antibodies in the hepatic CE samples for proteins of 24, 32 34, 44-46 and 52-54 kDa in molecular weight in 78.4%, 75.7%, 78.4% and 89.2% of the patients, respectively. In the pulmonary CE samples sera WB test enabled the detection of antibodies 24, 44-46, 100, 110, 116 and 120 124 kDa in molecular weight in 81.3%, 75.0%, 87.5%, 71.9%, 84.4% and 65.6% of the patients, respectively. We indicated that the antigenic components of high molecular weight can be good candidates for differentiation of hepatic CE from pulmonary CE.

Measuring the Effective Sampling Area of a Pheromone Trap for Monitoring Population Density of Southern Pine Beetle (Coleoptera: Scolytidae)

Treesearch

Peter Turchin; Francois J. Odendaal

1996-01-01

Multifunnel traps baited with frontalin and turpentine have been used to investigate dispersal of southern pine beetles, Dendroctonus front Zimmermann, and are routinely used in the southern United States to monitor population trends of this serious forest pest. However, there is no quantitative data on the effective sampling area of these traps that would allow us to...

Preparing Tomorrow's Administrators: A Quantitative Correlation Study of the Relationship between Emotional Intelligence and Effective Leadership Practices

ERIC Educational Resources Information Center

May-Vollmar, Kelly

2017-01-01

Purpose: The purpose of this quantitative correlation study was to identify whether there is a relationship between emotional intelligence and effective leadership practices, specifically with school administrators in Southern California K-12 public schools. Methods: This study was conducted using a quantitative descriptive design, correlation…

An Alu-based, MGB Eclipse real-time PCR method for quantitation of human DNA in forensic samples.

PubMed

Nicklas, Janice A; Buel, Eric

2005-09-01

The forensic community needs quick, reliable methods to quantitate human DNA in crime scene samples to replace the laborious and imprecise slot blot method. A real-time PCR based method has the possibility of allowing development of a faster and more quantitative assay. Alu sequences are primate-specific and are found in many copies in the human genome, making these sequences an excellent target or marker for human DNA. This paper describes the development of a real-time Alu sequence-based assay using MGB Eclipse primers and probes. The advantages of this assay are simplicity, speed, less hands-on-time and automated quantitation, as well as a large dynamic range (128 ng/microL to 0.5 pg/microL).

A Streamlined Western Blot Exercise: An Efficient and Greener Approach in the Laboratory Classroom

ERIC Educational Resources Information Center

Ness, Traci L.; Robinson, Rebekah L.; Mojadedi, Wais; Peavy, Lydia; Weiland, Mitch H.

2015-01-01

SDS-PAGE and western blotting are two commonly taught protein detection techniques in biochemistry and molecular biology laboratory classrooms. A pitfall associated with incorporating these techniques into the laboratory is the significant wait times that do not allow students to obtain timely results. The waiting associated with SDS-PAGE comes…

Optical properties of acute kidney injury measured by quantitative phase imaging

PubMed Central

Ban, Sungbea; Min, Eunjung; Baek, Songyee; Kwon, Hyug Moo; Popescu, Gabriel

2018-01-01

The diagnosis of acute kidney disease (AKI) has been examined mainly by histology, immunohistochemistry and western blot. Though these approaches are widely accepted in the field, it has an inherent limitation due to the lack of high-throughput and quantitative information. For a better understanding of prognosis in AKI, we present a new approach using quantitative phase imaging combined with a wide-field scanning platform. Through the phase-delay information from the tissue, we were able to predict a stage of AKI based on various optical properties such as light scattering coefficient and anisotropy. These optical parameters quantify the deterioration process of the AKI model of tissue. Our device would be a very useful tool when it is required to deliver fast feedback of tissue pathology or when diseases are related to mechanical properties such as fibrosis. PMID:29541494

Shallow Carbon Export from an Iron fertilised Plankton Bloom in the Southern Ocean

NASA Astrophysics Data System (ADS)

Sanders, R.; Pollard, R.; Morris, P.; Statham, P.; Moore, C. M. M.; Lucas, M.

2009-04-01

Some regions of the global ocean, notably the Southern Ocean, have high levels of macronutrients yet low levels of chlorophyll (the high nutrient, low chlorophyll or HNLC condition). Numerous artificial iron fertilization experiments conducted in the Southern Ocean have resulted in enhanced phytoplankton biomass and macronutrient drawdown. However the subsequent long-term biogeochemical consequences of such iron fertilization are unclear due in part to the limited size and duration of such experiments. An alternative way to assess the affect of iron over the Southern Ocean biological carbon pump is to observe the evolution of plankton production in regions of the Southern Ocean where shallow topography and Ocean currents interact to promote to release terrestrial iron into HNLC waters. During 2004-5 RRS Discovery conduced a complex programme of observations in such a region around the Crozet Islands in the SW Indian Ocean. The results of this programme, focussing on a quantitative estimate of carbon export per unit iron addition, will be presented.

Dot-Blot Immunoassay of Fasciola gigantica Infection using 27 kDa and Adult Worm Regurge Antigens in Egyptian Patients

PubMed Central

Kamel, Hanan H.; Saad, Ghada A.

2013-01-01

The purpose of the present study was to evaluate the potential role of the 27-Kilodalton (KDa) antigen versus Fasciola gigantica adult worm regurge antigens in a DOT-Blot assay and to assess this assay as a practical tool for diagnosis fascioliasis in Egyptian patients. Fasciola gigantica antigen of an approximate molecular mass 27-(KDa) was obtained from adult worms by a simple elution SDS-PAGE. A Dot-Blot was developed comparatively to adult worm regurge antigens for the detection of specific antibodies from patients infected with F. gigantica in Egypt. Control sera were obtained from patients with other parasitic infections and healthy volunteers to assess the test and compare between the antigens. The sensitivity, specificity, positive and negative predictive values of Dot-Blot using the adult worm regurge were 80%, 90%, 94.1%, and 69.2% respectively, while those using 27-KDa were 100% which confirms the diagnostic potential of this antigen. All patients infected with Fasciola were positive, with cross reactivity reported with Schistosoma mansoni serum samples. This 27-KDa Dot-Blot assay showed to be a promising test which can be used for serodiagnosis of fascioliasis in Egyptian patients especially, those presenting with hepatic disease. It is specific, sensitive and easy to perform method for the rapid diagnosis particularly when more complex laboratory tests are unavailable. PMID:23710084

Dot-blot immunoassay of Fasciola gigantica infection using 27 kDa and adult worm regurge antigens in Egyptian patients.

PubMed

Kamel, Hanan H; Saad, Ghada A; Sarhan, Rania M

2013-04-01

The purpose of the present study was to evaluate the potential role of the 27-Kilodalton (KDa) antigen versus Fasciola gigantica adult worm regurge antigens in a DOT-Blot assay and to assess this assay as a practical tool for diagnosis fascioliasis in Egyptian patients. Fasciola gigantica antigen of an approximate molecular mass 27-(KDa) was obtained from adult worms by a simple elution SDS-PAGE. A Dot-Blot was developed comparatively to adult worm regurge antigens for the detection of specific antibodies from patients infected with F. gigantica in Egypt. Control sera were obtained from patients with other parasitic infections and healthy volunteers to assess the test and compare between the antigens. The sensitivity, specificity, positive and negative predictive values of Dot-Blot using the adult worm regurge were 80%, 90%, 94.1%, and 69.2% respectively, while those using 27-KDa were 100% which confirms the diagnostic potential of this antigen. All patients infected with Fasciola were positive, with cross reactivity reported with Schistosoma mansoni serum samples. This 27-KDa Dot-Blot assay showed to be a promising test which can be used for serodiagnosis of fascioliasis in Egyptian patients especially, those presenting with hepatic disease. It is specific, sensitive and easy to perform method for the rapid diagnosis particularly when more complex laboratory tests are unavailable.

Identification of quantitative trait Loci for resistance to southern leaf blight and days to anthesis in a maize recombinant inbred line population.

PubMed

Balint-Kurti, P J; Krakowsky, M D; Jines, M P; Robertson, L A; Molnár, T L; Goodman, M M; Holl, J B

2006-10-01

ABSTRACT A recombinant inbred line population derived from a cross between the maize lines NC300 (resistant) and B104 (susceptible) was evaluated for resistance to southern leaf blight (SLB) disease caused by Cochliobolus heterostrophus race O and for days to anthesis in four environments (Clayton, NC, and Tifton, GA, in both 2004 and 2005). Entry mean and average genetic correlations between disease ratings in different environments were high (0.78 to 0.89 and 0.9, respectively) and the overall entry mean heritability for SLB resistance was 0.89. When weighted mean disease ratings were fitted to a model using multiple interval mapping, seven potential quantitative trait loci (QTL) were identified, the two strongest being on chromosomes 3 (bin 3.04) and 9 (bin 9.03-9.04). These QTL explained a combined 80% of the phenotypic variation for SLB resistance. Some time-point-specific SLB resistance QTL were also identified. There was no significant correlation between disease resistance and days to anthesis. Six putative QTL for time to anthesis were identified, none of which coincided with any SLB resistance QTL.

Immuno-Northern Blotting: Detection of RNA Modifications by Using Antibodies against Modified Nucleosides.

PubMed

Mishima, Eikan; Jinno, Daisuke; Akiyama, Yasutoshi; Itoh, Kunihiko; Nankumo, Shinnosuke; Shima, Hisato; Kikuchi, Koichi; Takeuchi, Yoichi; Elkordy, Alaa; Suzuki, Takehiro; Niizuma, Kuniyasu; Ito, Sadayoshi; Tomioka, Yoshihisa; Abe, Takaaki

2015-01-01

The biological roles of RNA modifications are still largely not understood. Thus, developing a method for detecting RNA modifications is important for further clarification. We developed a method for detecting RNA modifications called immuno-northern blotting (INB) analysis and herein introduce its various capabilities. This method involves the separation of RNAs using either polyacrylamide or agarose gel electrophoresis, followed by transfer onto a nylon membrane and subsequent immunoblotting using antibodies against modified nucleosides for the detection of specific modifications. We confirmed that INB with the antibodies for 1-methyladenosine (m1A), N6-methyladenosine (m6A), pseudouridine, and 5-methylcytidine (m5C) showed different modifications in a variety of RNAs from various species and organelles. INB with the anti-m5C antibody revealed that the antibody cross-reacted with another modification on DNA, suggesting the application of this method for characterization of the antibody for modified nucleosides. Additionally, using INB with the antibody for m1A, which is a highly specific modification in eukaryotic tRNA, we detected tRNA-derived fragments known as tiRNAs under the cellular stress response, suggesting the application for tracking target RNA containing specific modifications. INB with the anti-m6A antibody confirmed the demethylation of m6A by the specific demethylases fat mass and obesity-associated protein (FTO) and ALKBH5, suggesting its application for quantifying target modifications in separated RNAs. Furthermore, INB demonstrated that the knockdown of FTO and ALKBH5 increased the m6A modification in small RNAs as well as in mRNA. The INB method has high specificity, sensitivity, and quantitative capability, and it can be employed with conventional experimental apparatus. Therefore, this method would be useful for research on RNA modifications and metabolism.

An assessment of optical properties of dissolved organic material as quantitative source indicators in the Santa Ana River basin, Southern California

USGS Publications Warehouse

Bergamaschi, Brian A.; Kalve, Erica; Guenther, Larry; Mendez, Gregory O.; Belitz, Kenneth

2005-01-01

The ability to rapidly, reliably, and inexpensively characterize sources of dissolved organic material (DOM) in watersheds would allow water management agencies to more quickly identify problems in water sources, and to more efficiently allocate water resources by, for example, permitting real-time identification of high-quality water suitable for ground-water recharge, or poor-quality water in need of mitigation. This study examined the feasibility of using easily measurable intrinsic optical properties' absorbance and fluorescence spectra, as quantitative indicators of DOM sources and, thus, a predictor of water quality. The study focused on the Santa Ana River Basin, in southern California, USA, which comprises an area of dense urban development and an area of intense dairy production. Base flow in the Santa Ana Basin is primarily tertiary treated wastewater discharge. Available hydrologic data indicate that urban and agricultural runoff degrades water quality during storm events by introducing pathogens, nutrients, and other contaminants, including significant amounts of DOM. These conditions provide the basis for evaluating the use of DOM optical properties as a tracer of DOM from different sources. Sample spectra representing four principal DOM sources were identified among all samples collected in 1999 on the basis of basin hydrology, and the distribution of spectral variability within all the sample data. A linear mixing model provided quantitative estimates of relative endmember contribution to sample spectra for monthly, storm, and diurnal samples. The spectral properties of the four sources (endmembers), Pristine Water, Wastewater, Urban Water, and Dairy Water, accounted for 94 percent of the variability in optical properties observed in the study, suggesting that all important DOM sources were represented. The scale and distribution of the residual spectra, that not explained by the endmembers, suggested that the endmember spectra selected did not

Influence of the Inherited Glucose-6-phosphate Dehydrogenase Deficiency on the Appearance of Neonatal Hyperbilirubinemia in Southern Croatia.

PubMed

Cherepnalkovski, Anet Papazovska; Marusic, Eugenija; Piperkova, Katica; Lozic, Bernarda; Skelin, Ana; Gruev, Todor; Krzelj, Vjekoslav

2015-10-01

Neonatal hyperbilirubinemia is a common clinical manifestation of the inherited glucose-6-phosphate dehydrogenase (G6PD) deficiency. The aim of this study was to investigate the influence of the inherited G6PD deficiency on the appearance of neonatal hyperbilirubinemia in southern Croatia. The fluorescent spot test (FST) was used in a retrospective study to screen blood samples of 513 male children who had neonatal hyperbilirubinemia, of unknown cause, higher than 240 μmol/L. Fluorescence readings were performed at the beginning and at the fifth and tenth minute of incubation and were classified into three groups bright fluorescence (BF), weak fluorescence (WF) and no fluorescence (NF). Normal samples show bright fluorescence. All NF and WF samples at the fifth minute were quantitatively measured using the spectrophotometric method. Bright fluorescence was present in 461 patients (89.9%) at the fifth minute. The remaining 52 (10.1%) were quantitatively estimated using the spectrophotometric method. G6PD deficiency was observed in 38 patients (7.4%). Prevalence rate of G6PD deficiency among male newborns with hyperbilirubinemia in southern Croatia is significantly higher (p < 0.01) compared with the previously reported prevalence rate among male in general population of southern Croatia (0.75%). We recommend FST to be performed in hyperbilirubinemic newborns in southern Croatia.

Influence of trap modifications and environmental predictors on capture success of southern flying squirrels

USGS Publications Warehouse

Jacques, Christopher N.; Zweep, James S.; Scheihing, Mary E.; Rechkemmer, Will T.; Jenkins, Sean E.; Klaver, Robert W.; Dubay, Shelli A.

2017-01-01

Sherman traps are the most commonly used live traps in studies of small mammals and have been successfully used in the capture of arboreal species such as the southern flying squirrel (Glaucomys volans). However, southern flying squirrels spend proportionately less time foraging on the ground, which necessitates above-ground trapping methods and modifications of capture protocols. Further, quantitative estimates of the factors affecting capture success of flying squirrel populations have focused solely on effects of trapping methodologies. We developed and evaluated the efficacy of a portable Sherman trap design for capturing southern flying squirrels during 2015–2016 at the Alice L. Kibbe Field Station, Illinois, USA. Additionally, we used logistic regression to quantify potential effects of time-dependent (e.g., weather) and time-independent (e.g., habitat, extrinsic) factors on capture success of southern flying squirrels. We recorded 165 capture events (119 F, 44 M, 2 unknown) using our modified Sherman trap design. Probability of capture success decreased 0.10/1° C increase in daily maximum temperature and by 0.09/unit increase (km/hr) in wind speed. Conversely, probability of capture success increased by 1.2/1° C increase in daily minimum temperature. The probability of capturing flying squirrels was negatively associated with trap orientation. When tree-mounted traps are required, our modified trap design is a safe, efficient, and cost-effective method of capturing animals when moderate weather (temp and wind speed) conditions prevail. Further, we believe that strategic placement of traps (e.g., northeast side of tree) and quantitative information on site-specific (e.g., trap location) characteristics (e.g., topographical features, slope, aspect, climatologic factors) could increase southern flying squirrel capture success. © 2017 The Wildlife Society.

Fire and human history of a barren-forest mosaic in Southern Indiana

Treesearch

Richard P. Guyette; Daniel C. Dey; Michael C. Stambaugh

2002-01-01

The purpose of this paper is to provide quantitative fire history information from a historically unique region, the oak barrens of the Interior Low Plateau Ecoregion. We sampled 27 post oak (Quercus stellata Wangenh.) trees from the Boone Creek watershed in southern Indiana. The period of tree-ring record ranged in calendar years from 1654 to 1999...

Blotting Assisted by Heating and Solvent Extraction for DESI-MS Imaging

NASA Astrophysics Data System (ADS)

Cabral, Elaine C.; Mirabelli, Mario F.; Perez, Consuelo J.; Ifa, Demian R.

2013-06-01

Imprints of potato sprout ( Solanum tuberosum L.), gingko leaves (Gingko biloba L. ) and strawberries (Fragaria x ananassa Duch. ) were successfully imaged by desorption electrospray ionization mass spectrometry (DESI-MS) on TLC plates through blotting assisted by heating and/or solvent extraction. Ion images showing the distribution of significant compounds such as glycoalkaloid toxins in potato sprout, ginkgolic acids and flavonoids in ginkgo leaves, and sugars and anthocyanidin in strawberry were obtained. Practical implications of this work include analysis of a wide range of irregular or soft materials by different imprinting conditions without requiring the addition of matrices or use of specific kinds of surfaces.

Evaluation of Rice Resistance to Southern Rice Black-Streaked Dwarf Virus and Rice Ragged Stunt Virus through Combined Field Tests, Quantitative Real-Time PCR, and Proteome Analysis.

PubMed

Wang, Zhenchao; Yu, Lu; Jin, Linhong; Wang, Wenli; Zhao, Qi; Ran, Longlu; Li, Xiangyang; Chen, Zhuo; Guo, Rong; Wei, Yongtian; Yang, Zhongcheng; Liu, Enlong; Hu, Deyu; Song, Baoan

2017-02-22

Diseases caused by southern rice black-streaked dwarf virus (SRBSDV) and rice ragged stunt virus (RRSV) considerably decrease grain yield. Therefore, determining rice cultivars with high resistance to SRBSDV and RRSV is necessary. In this study, rice cultivars with high resistance to SRBSDV and RRSV were evaluated through field trials in Shidian and Mangshi county, Yunnan province, China. SYBR Green I-based quantitative real-time polymerase chain reaction (qRT-PCR) analysis was used to quantitatively detect virus gene expression levels in different rice varieties. The following parameters were applied to evaluate rice resistance: acre yield (A.Y.), incidence of infected plants (I.I.P.), virus load (V.L.), disease index (D.I.), and insect quantity (I.Q.) per 100 clusters. Zhongzheyou1 (Z1) and Liangyou2186 (L2186) were considered the most suitable varieties with integrated higher A.Y., lower I.I.P., V.L., D.I. and I.Q. In order to investigate the mechanism of rice resistance, comparative label-free shotgun liquid chromatography tandem-mass spectrometry (LC-MS/MS) proteomic approaches were applied to comprehensively describe the proteomics of rice varieties' SRBSDV tolerance. Systemic acquired resistance (SAR)-related proteins in Z1 and L2186 may result in the superior resistance of these varieties compared with Fengyouxiangzhan (FYXZ).

Developing Quantitative Reasoning: Will Taking Traditional Math Courses Suffice? An Empirical Study

ERIC Educational Resources Information Center

Agustin, Ma Zenia; Agustin, Marcus; Brunkow, Paul; Thomas, Susan

2012-01-01

Southern Illinois University Edwardsville (SIUE) is a postbaccalaureate comprehensive university in the Midwest. In 2005, SIUE embarked on a challenging journey of general education reform. A review of the current general education program revealed that it is possible for a student to graduate from SIUE without taking a quantitative course. Hence,…

Multi-proxy palaeoclimate reconstructions from peatlands in southern South America

NASA Astrophysics Data System (ADS)

Roland, Thomas; Hughes, Paul; Mauquoy, Dmitri; van Bellen, Simon; Daley, Tim; Loader, Neil; Street-Perrott, Alayne

2014-05-01

There is a relative paucity of palaeoclimatic archives in South America relative to many other regions of the world. This paucity must be addressed in order to validate climate models and improve our understanding of the global climate system. The southern westerlies represent an important component of climatic variability in the region and, in turn, their migration and changes in their intensity can play a key role in determining whether the Southern Ocean functions as a sink or source of atmospheric carbon dioxide. Increased ventilation of deep waters with elevated concentrations of dissolved inorganic carbon, driven by enhanced Ekman transport, leads to increased outgassing of carbon dioxide. However, as instrumental records are limited to the latter half of the twentieth century, little is known about the long-term variability of the southern Westerlies and their subsequent effects. The Peninsula Brunswick and Isla Grande de Tierra del Fuego are directly situated in the core path of the southern westerlies during the Austral summer and they are ideally suited for studies of past variability in westerly intensity and position. The region's abundant peatlands are capable of recording these long-term changes, as wind intensity and westerly position affects precipitation and temperature, two key drivers (i.e. P-E) of water-table dynamics in ombrotrophic peatlands. Currently, the peatlands of southern Patagonia represent a relatively unexploited resource in terms of palaeoclimate reconstruction. As a result, we have developed a new regional network of multi-proxy (testate amoebae, plant macrofossils, stable isotopes) archives, supported by high-resolution radiocarbon chronologies, to develop quantitative climate reconstructions for southern South America spanning the last ~2000 years using Sphagnum magellanicum-dominated peat deposits.

Detection of Diverse and High Molecular Weight Nesprin-1 and Nesprin-2 Isoforms Using Western Blotting.

PubMed

Carthew, James; Karakesisoglou, Iakowos

2016-01-01

Heavily utilized in cell and molecular biology, western blotting is considered a crucial technique for the detection and quantification of proteins within complex mixtures. In particular, the detection of members of the nesprin (nuclear envelope spectrin repeat protein) family has proven difficult to analyze due to their substantial isoform diversity, molecular weight variation, and the sheer size of both nesprin-1 and nesprin-2 giant protein variants (>800 kDa). Nesprin isoforms contain distinct domain signatures, perform differential cytoskeletal associations, occupy different subcellular compartments, and vary in their tissue expression profiles. This structural and functional variance highlights the need to distinguish between the full range of proteins within the nesprin protein family, allowing for greater understanding of their specific roles in cell biology and disease. Herein, we describe a western blotting protocol modified for the detection of low to high molecular weight (50-1000 kDa) nesprin proteins.

Quantitative assessment of direct and indirect landslide risk along transportation lines in southern India

NASA Astrophysics Data System (ADS)

Jaiswal, P.; van Westen, C. J.; Jetten, V.

2010-06-01

A quantitative approach for landslide risk assessment along transportation lines is presented and applied to a road and a railway alignment in the Nilgiri hills in southern India. The method allows estimating direct risk affecting the alignments, vehicles and people, and indirect risk resulting from the disruption of economic activities. The data required for the risk estimation were obtained from historical records. A total of 901 landslides were catalogued initiating from cut slopes along the railway and road alignment. The landslides were grouped into three magnitude classes based on the landslide type, volume, scar depth, run-out distance, etc and their probability of occurrence was obtained using frequency-volume distribution. Hazard, for a given return period, expressed as the number of landslides of a given magnitude class per kilometre of cut slopes, was obtained using Gumbel distribution and probability of landslide magnitude. In total 18 specific hazard scenarios were generated using the three magnitude classes and six return periods (1, 3, 5, 15, 25, and 50 years). The assessment of the vulnerability of the road and railway line was based on damage records whereas the vulnerability of different types of vehicles and people was subjectively assessed based on limited historic incidents. Direct specific loss for the alignments (railway line and road), vehicles (train, bus, lorry, car and motorbike) was expressed in monetary value (US), and direct specific loss of life of commuters was expressed in annual probability of death. Indirect specific loss (US) derived from the traffic interruption was evaluated considering alternative driving routes, and includes losses resulting from additional fuel consumption, additional travel cost, loss of income to the local business, and loss of revenue to the railway department. The results indicate that the total loss, including both direct and indirect loss, from 1 to 50 years return period, varies from US 90 840 to US

Intersection of Southern Parkway and Southern Heights, looking toward the ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Intersection of Southern Parkway and Southern Heights, looking toward the Beechmont Historic District, showing changes in landscaping, northeast - Southern Heights-Beechmont District Landscapes, Louisville, Jefferson County, KY

Frictional strength and heat flow of southern San Andreas Fault

NASA Astrophysics Data System (ADS)

Zhu, P. P.

2016-01-01

Frictional strength and heat flow of faults are two related subjects in geophysics and seismology. To date, the investigation on regional frictional strength and heat flow still stays at the stage of qualitative estimation. This paper is concentrated on the regional frictional strength and heat flow of the southern San Andreas Fault (SAF). Based on the in situ borehole measured stress data, using the method of 3D dynamic faulting analysis, we quantitatively determine the regional normal stress, shear stress, and friction coefficient at various seismogenic depths. These new data indicate that the southern SAF is a weak fault within the depth of 15 km. As depth increases, all the regional normal and shear stresses and friction coefficient increase. The former two increase faster than the latter. Regional shear stress increment per kilometer equals 5.75 ± 0.05 MPa/km for depth ≤15 km; regional normal stress increment per kilometer is equal to 25.3 ± 0.1 MPa/km for depth ≤15 km. As depth increases, regional friction coefficient increment per kilometer decreases rapidly from 0.08 to 0.01/km at depths less than ~3 km. As depth increases from ~3 to ~5 km, it is 0.01/km and then from ~5 to 15 km, and it is 0.002/km. Previously, frictional strength could be qualitatively determined by heat flow measurements. It is difficult to obtain the quantitative heat flow data for the SAF because the measured heat flow data exhibit large scatter. However, our quantitative results of frictional strength can be employed to investigate the heat flow in the southern SAF. We use a physical quantity P f to describe heat flow. It represents the dissipative friction heat power per unit area generated by the relative motion of two tectonic plates accommodated by off-fault deformation. P f is called "fault friction heat." On the basis of our determined frictional strength data, utilizing the method of 3D dynamic faulting analysis, we quantitatively determine the regional long-term fault

Quantifying dispersal of southern pine beetles with mark-recapture experiments and a diffusion model

Treesearch

P. Turchin; W.T. Thoeny

1993-01-01

Pest management decisions should take into consideration quantitative information on dispersal of insect pests, but such information is often lacking.The goal of this study was to measure intraforest dispersal in the southern pine beetle (SPB).We developed an analytical formula for interpreting data from mark-recapture studies of insect dispersal.The proposed...

A homozygous transthyretin variant associated with senile systemic amyloidosis: evidence for a late-onset disease of genetic etiology.

PubMed Central

Jacobson, D R; Gorevic, P D; Buxbaum, J N

1990-01-01

Senile systemic amyloidosis (SSA) is a late-onset disease characterized by deposition of amyloid fibrils containing transthyretin (TTR). Amino acid sequencing of protein isolated from the amyloid fibrils of a patient with SSA identified TTR containing a position - 122 isoleucine-for-valine substitution. This change led to the prediction of a genomic G-to-A transition, destroying an MaeIII restriction site. We confirmed the presence of the variant DNA fragment both by Southern blotting and by visualization of MaeIII digests of DNA amplified around codon 122, by using the polymerase chain reaction. The patient's DNA was entirely resistant to MaeIII cleavage; therefore, only the mutant sequence was present. DNA from none of either 24 controls or six other SSA patients contained the variant. Quantitative Southern blotting demonstrated that the patient's DNA contained two copies of the TTR gene per genome; the mutation was therefore homozygous rather than hemizygous. In the present case, the homozygous mutation TTR (122 Val----Ile) is associated with SSA, a finding which is consistent with autosomal recessive inheritance of this condition. Images Figure 2 Figure 4 Figure 5 Figure 6 Figure 7 PMID:2349941

In vitro detection of canine distemper virus nucleic acid with a virus-specific cDNA probe by dot-blot and in situ hybridization.

PubMed

Oglesbee, M; Jackwood, D; Perrine, K; Axthelm, M; Krakowka, S; Rice, J

1986-11-01

A cDNA library was prepared from canine distemper viral (CDV) messenger RNA (mRNA) derived from Vero cells lytically infected with the Onderstepoort strain (Ond) of CDV. A 300 base pair insert was identified which, by Northern blot analysis and Sanger sequence data, was shown to be specific to the nucleocapsid gene. The nucleocapsid (NC) clone was radiolabelled with 32P using nick translation and used to detect viral RNA in both dot-blot and in situ preparations of Vero cells lytically infected with Onderstepoort CDV (Ond-CDV) and immortalized mink lung cells persistently infected with racoon origin CDV (CCL64-RCDV). Dot-blot hybridization results paralleled immunofluorescent results in the lytically infected cells. In 18 persistently infected cell lines from the RCDV-CCL64 parental stock, 13 lines were positive and two were negative on both immunofluorescence and dot-blot hybridization analysis for CDV antigen and RNA, respectively. Viral nucleic acid was detected in these persistently infected cells, where as few as 1.9% of the members of a line were positive on immunofluorescence. A dot-blot autoradiographic signal was obtained in three lines which were negative for CDV antigen. CDV RNA was detected in both lytically and persistently infected cell lines by in situ hybridization, where decreasing probe length was important in increasing the sensitivity of this assay. Viral RNA was detected in over 90% of the lytically infected cells, where only 70% were positive for viral antigen by immunofluorescence.

Quantitative Structure-Cytotoxicity Relationship of Cinnamic Acid Phenetyl Esters.

PubMed

Uesawa, Yoshihiro; Sakagami, Hiroshi; Okudaira, Noriyuki; Toda, Kazuhiro; Takao, Koichi; Kagaya, Hajime; Sugita, Yoshiaki

2018-02-01

Many phenolic acid phenethyl esters possess diverse biological effects including antioxidant, cytoprotective, anti-inflammation and anti-tumor activities. However, most previous antitumor studies have not considered the cytotoxicity against normal cells. Ten cinnamic acid phenetyl esters were subjected to quantitative structure-activity relationship (QSAR) analysis, based on their cytotoxicity and tumor-specificity, in order to find their new biological activities. Cytotoxicity against four human oral squamous cell carcinoma cell lines and three oral normal mesenchymal cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Tumor specificity (TS) was evaluated by the ratio of the mean 50% cytotoxic concentration (CC 50 ) against normal oral cells to that against human oral squamous cell carcinoma cell lines. Potency-selectivity expression (PSE) value was calculated by dividing the TS value by CC 50 against tumor cells. Apoptosis markers were detected by western blot analysis. Physicochemical, structural and quantum-chemical parameters were calculated based on the conformations optimized by force-field minimization. Western blot analysis demonstrated that [ 9 ] stimulated the cleavage of caspase-3, suggesting the induction of apoptosis. QSAR analysis demonstrated that TS values were correlated with shape, size and ionization potential. Chemical modification of the lead compound may be a potential choice for designing a new type of anticancer drugs. Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

A gene encoding maize caffeoyl-CoA O-methyltransferase confers quantitative resistance to multiple pathogens.

PubMed

Yang, Qin; He, Yijian; Kabahuma, Mercy; Chaya, Timothy; Kelly, Amy; Borrego, Eli; Bian, Yang; El Kasmi, Farid; Yang, Li; Teixeira, Paulo; Kolkman, Judith; Nelson, Rebecca; Kolomiets, Michael; L Dangl, Jeffery; Wisser, Randall; Caplan, Jeffrey; Li, Xu; Lauter, Nick; Balint-Kurti, Peter

2017-09-01

Alleles that confer multiple disease resistance (MDR) are valuable in crop improvement, although the molecular mechanisms underlying their functions remain largely unknown. A quantitative trait locus, qMdr 9.02 , associated with resistance to three important foliar maize diseases-southern leaf blight, gray leaf spot and northern leaf blight-has been identified on maize chromosome 9. Through fine-mapping, association analysis, expression analysis, insertional mutagenesis and transgenic validation, we demonstrate that ZmCCoAOMT2, which encodes a caffeoyl-CoA O-methyltransferase associated with the phenylpropanoid pathway and lignin production, is the gene within qMdr 9.02 conferring quantitative resistance to both southern leaf blight and gray leaf spot. We suggest that resistance might be caused by allelic variation at the level of both gene expression and amino acid sequence, thus resulting in differences in levels of lignin and other metabolites of the phenylpropanoid pathway and regulation of programmed cell death.

Monoclonal Antibodies against Small Molecule Natural Products and Their Applications, Eastern Blotting and Knockout Extract

PubMed Central

Shoyama, Yukihiro

2011-01-01

To determine the hapten number in hapten-carrier protein conjugate matrix-assisted laser desorption/ionization (MALDI) tof mass spectrometry was applied. Highly specific anti-ginsenoside Rb1 and Rg1 monoclonal antibodies (MAbs) were prepared. Ginsenosides were developed on thin layer chromatography (TLC) plates which were covered by a polyvinylidene difluoride (PVDF) membrane resulting in blotting. The membrane was treated with NaIO4 solution to release the aldehyde group on the sugar moiety of the ginsenosides. By treatment of the membrane with a protein solution the ginsenoside-protein conjugation as a Schiff-base occurred, which can function to fix it to the PVDF membrane. A part of the ginsenoside aglycone was reacted with anti-ginsenoside Rb1 MAb, secondary MAb conjugated with enzyme and finally a substrate was added, resulting in a specific and highly sensitive staining that we named Eastern blotting. Furthermore, it makes one-step isolation of ginsenoside Rb1 possible using an immuno-affinity column conjugated with anti-ginsenoside Rb1 MAb. Furthermore, immunoaffinity concentration was carried out allowing high sensitivity analysis of lower concentrations of ginsenoside Rb1 so that several unknown bands could be structurally determined.

Western blot (immunoblot) assay of small, round-structured virus associated with an acute gastroenteritis outbreak in Tokyo.

PubMed

Hayashi, Y; Ando, T; Utagawa, E; Sekine, S; Okada, S; Yabuuchi, K; Miki, T; Ohashi, M

1989-08-01

Small, round-structured virus (SRSV) was detected in a stool specimen of a patient during an acute gastroenteritis outbreak in Tokyo and was tentatively named SRSV-9. SRSV-9 was purified by sucrose velocity gradient centrifugation after CsCl density gradient centrifugation. The buoyant density of SRSV-9 appeared to be 1.36 g/ml in CsCl. A Western blot (immunoblot) assay using the biotin-avidin system revealed that SRSV-9 was antigenically related to the Hawaii agent but distinct from the Norwalk agent and contained a single major structural protein with a molecular size of 63.0 +/- 0.6 kilodaltons. The prevalence of SRSV-9 infection in Tokyo was surveyed by the Western blot antibody assay by using a crude virus preparation as the antigen. Seroconversion was observed in 56.5% of the patients involved in the outbreaks from which SRSV was detected by electron microscopy.

Establishment and application of a modified membrane-blot assay for Rhizomucor miehei lipases aimed at improving their methanol tolerance and thermostability.

PubMed

He, Dong; Luo, Wen; Wang, Zhiyuan; Lv, Pengmei; Yuan, Zhenhong; Huang, Shaowei; Xv, Jingliang

2017-07-01

Directed evolution has been proved an effective way to improve the stability of proteins, but high throughput screening assays for directed evolution with simultaneous improvement of two or more properties are still rare. In this study, we aimed to establish a membrane-blot assay for use in the high-throughput screening of Rhizomucor miehei lipases (RMLs). With the assistance of the membrane-blot screening assay, a mutant E47K named G10 that showed improved thermal stability was detected in the first round of error-prone PCR. Using G10 as the parent, two variants G10-11 and G10-20 that showed improved thermal stability and methanol tolerance without loss of activity compared to the wild type RML were obtained. The T 50 60 -value of G10-11 and G10-20 increased by 12°C and 6.5°C, respectively. After incubation for 1h, the remaining residual activity of G10-11 and G10-20 was 63.45% and 74.33%, respectively, in 50% methanol, and 15.98% and 30.22%, respectively, in 80% methanol. Thus, we successfully developed a membrane-blot assay that could be used for the high-throughput screening of RMLs with improved thermostability and methanol tolerance. Based on our findings, we believe that our newly developed membrane-blot assay will have potential applications in directed evolution in the future. Copyright © 2017 Elsevier Inc. All rights reserved.

Models of Academic Governance and Institutional Power in Southern Baptist Related Liberal Arts Colleges and Universities

ERIC Educational Resources Information Center

Garrison, Michael Shane

2010-01-01

The purpose of this descriptive-quantitative study was to examine which models of academic governance are utilized by Southern Baptist related liberal arts colleges and universities. Special attention was given to the distribution of institutional power among seventeen campus leadership groups or power holders. Using J. Victor Baldridge's models…

A preliminary test of estimating forest site quality using species composition in a southern Appalachian watershed

Treesearch

W. Henry McNab; David L. Loftis

2013-01-01

Characteristic arborescent communities of mesophytic or xerophytic species have long been recognized as indicative of forest site quality in the Southern Appalachians, where soil moisture availability is the primary environmental variable affecting productivity. But, a workable quantitative system of site classification based on species composition is not available. We...

Southern limits of distribution of the intertidal gobies Chaenogobius annularis and C. gulosus support the existence of a biogeographic boundary in southern Japan (Teleostei, Perciformes, Gobiidae)

PubMed Central

Murase, Atsunobu; Miki, Ryohei; Motomura, Hiroyuki

2017-01-01

Abstract Understanding the distributional patterns of individual animal groups with respect to coastal topology and the local physical environment provides essential foundational frameworks for marine zoogeography. In the northwestern Pacific waters of Japan, the distributional pattern of some cool-temperate species of marine fishes suggests the existence of a biogeographic boundary corresponding to a long sandy shore on the eastern coast of Kyushu, southern Japan. The existence of this hypothetical biogeographic boundary was tested by mapping the southern distributional limit of two species of cool-temperate intertidal gobies, Chaenogobius annularis and C. gulosus, which are endemic to East Asia and common in rock pools within their range in the Japanese Archipelago. Distribution and abundance were assessed by survey of museum collections from south-east Kyushu (i.e., the entire coasts of Kagoshima and Miyazaki prefectures); and a quantitative survey of the abundance of these gobies in rock pools at various sites around the hypothesized boundary on the eastern coast of Kyushu, including the subtropical Tanega-shima Island. The museum collection survey showed different distribution patterns between the two species: C. annularis was distributed along the entire coasts of south-east Kyushu including subtropical islands, whereas C. gulosus was distributed along these coasts, including one site on a subtropical island, except for an area south of the hypothesized boundary on the eastern coast of Kyushu. The density and occurrence rates of C. annularis in rock pools decreased with latitude, it being absent from a subtropical island, and C. gulosus was not detected from sites south of the hypothesized boundary. The qualitative survey showed that the southernmost records of C. annularis and C. gulosus were the adjacent subtropical islands (Yaku-shima and Tanega-shima islands respectively), although the quantitative survey suggested that their normal range of distribution

A rop net and removable walkway used to quantitatively sample fishes over wetland surfaces in the dwarf mangrove of the Southern Everglades

USGS Publications Warehouse

Lorenz, J.J.; McIvor, C.C.; Powell, G.V.N.; Frederick, P.C.

1997-01-01

We describe a 9 m2 drop net and removable walkways designed to quantify densities of small fishes in wetland habitats with low to moderate vegetation density. The method permits the collection of small, quantitative, discrete samples in ecologically sensitive areas by combining rapid net deployment from fixed sites with the carefully contained use of the fish toxicant rotenone. This method requires very little contact with the substrate, causes minimal alteration to the habitat being sampled, samples small fishes in an unbiased manner, and allows for differential sampling of microhabitats within a wetland. When used in dwarf red mangrove (Rhizophora mangle) habitat in southern Everglades National Park and adjacent areas (September 1990 to March 1993), we achieved high recovery efficiencies (78–90%) for five common species <110 mm in length. We captured 20,193 individuals of 26 species. The most abundant fishes were sheepshead minnowCyprinodon variegatus, goldspotted killifishFloridichthys carpio, rainwater killifishLucania parva, sailfin mollyPoecilia latipinna, and the exotic Mayan cichlidCichlasoma urophthalmus. The 9 m2 drop net and associated removable walkways are versatile and can be used in a variety of wetland types, including both interior and coastal wetlands with either herbaceous or woody vegetation.

Mixed-methods quantitative-qualitative study of 29 nonagenarians and centenarians in rural Southern Italy: focus on positive psychological traits.

PubMed

Scelzo, Anna; Di Somma, Salvatore; Antonini, Paola; Montross, Lori P; Schork, Nicholas; Brenner, David; Jeste, Dilip V

2018-01-01

This was a study of positive psychological traits in a group of rural Italians aged 90 to 101 years, and their children or other family members. Mixed-methods quantitative (standardized rating scales) and qualitative (semi-structured interviews) study. Study participants' homes in nine villages in the Cilento region of southern Italy. Twenty-nine nonagenarians and centenarians and 51 family members aged 51-75 years, selected by their general practitioners as a part of a larger study called CIAO (Cilento Initiative on Aging Outcomes). We used published rating scales of mental and physical well-being, resilience, optimism, anxiety, depression, and perceived stress. Qualitative interviews gathered personal narratives of the oldest-old individuals, including migrations, traumatic events, and beliefs. Family members described their impressions about the personality traits of their older relative. Participants age ≥90 years had worse physical health but better mental well-being than their younger family members. Mental well-being correlated negatively with levels of depression and anxiety in both the groups. The main themes that emerged from qualitative interviews included positivity (resilience and optimism), working hard, and bond with family and religion, as described in previously published studies of the oldest old, but also a need for control and love of the land, which appeared to be unique features of this rural population. Exceptional longevity was characterized by a balance between acceptance of and grit to overcome adversities along with a positive attitude and close ties to family, religion, and land, providing purpose in life.

Evaluation of the Western blotting method for the diagnosis of congenital toxoplasmosis.

PubMed

Capobiango, Jaqueline Dario; Monica, Thaís Cabral; Ferreira, Fernanda Pinto; Mitsuka-Breganó, Regina; Navarro, Italmar Teodorico; Garcia, João Luis; Reiche, Edna Maria Vissoci

To evaluate the Western blotting method for the detection of IgG anti-Toxoplasma gondii (T. gondii) (IgG-WB) in the serum of children with suspected congenital toxoplasmosis. We accompanied 47 mothers with acquired toxoplasmosis in pregnancy and their children, between June of 2011 and June of 2014. The IgG-WB was done in house and the test was considered positive if the child had antibodies that recognized at least one band on IgG blots different from the mother's or with greater intensity than the corresponding maternal band, during the first three months of life. 15 children (15.1%) met the criteria for congenital toxoplasmosis and 32 (32.3%) had the diagnosis excluded. The symptoms were observed in 12 (80.0%) children and the most frequent were cerebral calcification in 9 (60.0%), chorioretinitis in 8 (53.3%), and hydrocephalus in 4 (26.6%). IgM antibodies anti-T. gondii detected by chemiluminescence (CL) were found in 6 (40.0%) children and the polymerase chain reaction (PCR) for detection of T. gondii DNA was positive in 5 of 7 performed (71.4%). The sensitivity of IgG-WB was of 60.0% [95% confidence interval (CI) 32.3-83.7%] and specificity 43.7% (95% CI 26.7-62.3%). The sensitivity of IgG-WB increased to 76.0 and 89.1% when associated to the research of IgM anti-T. gondii or PCR, respectively. The IgG-WB showed greater sensitivity than the detection of IgM anti-T. gondii; therefore, it can be used for the diagnosis of congenital toxoplasmosis in association with other congenital infection markers. Copyright © 2016 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.

Southern California Disasters II

NASA Technical Reports Server (NTRS)

Nicholson, Heather; Todoroff, Amber L.; LeBoeuf, Madeline A.

2015-01-01

The USDA Forest Service (USFS) has multiple programs in place which primarily utilize Landsat imagery to produce burn severity indices for aiding wildfire damage assessment and mitigation. These indices provide widely-used wildfire damage assessment tools to decision makers. When the Hyperspectral Infrared Imager (HyspIRI) is launched in 2022, the sensor's hyperspectral resolution will support new methods for assessing natural disaster impacts on ecosystems, including wildfire damage to forests. This project used simulated HyspIRI data to study three southern California fires: Aspen, French, and King. Burn severity indices were calculated from the data and the results were quantitatively compared to the comparable USFS products currently in use. The final results from this project illustrate how HyspIRI data may be used in the future to enhance assessment of fire-damaged areas and provide additional monitoring tools for decision support to the USFS and other land management agencies.

Reproducibility of telomere length assessment: an international collaborative study.

PubMed

Martin-Ruiz, Carmen M; Baird, Duncan; Roger, Laureline; Boukamp, Petra; Krunic, Damir; Cawthon, Richard; Dokter, Martin M; van der Harst, Pim; Bekaert, Sofie; de Meyer, Tim; Roos, Goran; Svenson, Ulrika; Codd, Veryan; Samani, Nilesh J; McGlynn, Liane; Shiels, Paul G; Pooley, Karen A; Dunning, Alison M; Cooper, Rachel; Wong, Andrew; Kingston, Andrew; von Zglinicki, Thomas

2015-10-01

Telomere length is a putative biomarker of ageing, morbidity and mortality. Its application is hampered by lack of widely applicable reference ranges and uncertainty regarding the present limits of measurement reproducibility within and between laboratories. We instigated an international collaborative study of telomere length assessment: 10 different laboratories, employing 3 different techniques [Southern blotting, single telomere length analysis (STELA) and real-time quantitative PCR (qPCR)] performed two rounds of fully blinded measurements on 10 human DNA samples per round to enable unbiased assessment of intra- and inter-batch variation between laboratories and techniques. Absolute results from different laboratories differed widely and could thus not be compared directly, but rankings of relative telomere lengths were highly correlated (correlation coefficients of 0.63-0.99). Intra-technique correlations were similar for Southern blotting and qPCR and were stronger than inter-technique ones. However, inter-laboratory coefficients of variation (CVs) averaged about 10% for Southern blotting and STELA and more than 20% for qPCR. This difference was compensated for by a higher dynamic range for the qPCR method as shown by equal variance after z-scoring. Technical variation per laboratory, measured as median of intra- and inter-batch CVs, ranged from 1.4% to 9.5%, with differences between laboratories only marginally significant (P = 0.06). Gel-based and PCR-based techniques were not different in accuracy. Intra- and inter-laboratory technical variation severely limits the usefulness of data pooling and excludes sharing of reference ranges between laboratories. We propose to establish a common set of physical telomere length standards to improve comparability of telomere length estimates between laboratories. © The Author 2014. Published by Oxford University Press on behalf of the International Epidemiological Association.

Proviral Features of Human T Cell Leukemia Virus Type 1 in Carriers with Indeterminate Western Blot Analysis Results

PubMed Central

Sekizuka, Tsuyoshi; Yamochi, Tadanori; Firouzi, Sanaz; Sato, Tomoo; Umeki, Kazumi; Sasaki, Daisuke; Hasegawa, Hiroo; Kubota, Ryuji; Sobata, Rieko; Matsumoto, Chieko; Kaneko, Noriaki; Momose, Haruka; Araki, Kumiko; Saito, Masumichi; Nosaka, Kisato; Utsunomiya, Atae; Koh, Ki-Ryang; Ogata, Masao; Uchimaru, Kaoru; Iwanaga, Masako; Sagara, Yasuko; Yamano, Yoshihisa; Okayama, Akihiko; Miura, Kiyonori; Satake, Masahiro; Saito, Shigeru; Itabashi, Kazuo; Yamaguchi, Kazunari; Kuroda, Makoto; Watanabe, Toshiki; Okuma, Kazu

2017-01-01

ABSTRACT Western blotting (WB) for human T cell leukemia virus type 1 (HTLV-1) is performed to confirm anti-HTLV-1 antibodies detected at the initial screening of blood donors and in pregnant women. However, the frequent occurrence of indeterminate results is a problem with this test. We therefore assessed the cause of indeterminate WB results by analyzing HTLV-1 provirus genomic sequences. A quantitative PCR assay measuring HTLV-1 provirus in WB-indeterminate samples revealed that the median proviral load was approximately 100-fold lower than that of WB-positive samples (0.01 versus 0.71 copy/100 cells). Phylogenic analysis of the complete HTLV-1 genomes of WB-indeterminate samples did not identify any specific phylogenetic groups. When we analyzed the nucleotide changes in 19 HTLV-1 isolates from WB-indeterminate samples, we identified 135 single nucleotide substitutions, composed of four types, G to A (29%), C to T (19%), T to C (19%), and A to G (16%). In the most frequent G-to-A substitution, 64% occurred at GG dinucleotides, indicating that APOBEC3G is responsible for mutagenesis in WB-indeterminate samples. Moreover, interestingly, five WB-indeterminate isolates had nonsense mutations in Pol and/or Tax, Env, p12, and p30. These findings suggest that WB-indeterminate carriers have low production of viral antigens because of a combination of a low proviral load and mutations in the provirus, which may interfere with host recognition of HTLV-1 antigens. PMID:28701419

Identification of recombinant human EPO variants in greyhound plasma and urine by ELISA, LC-MS/MS and western blotting: a comparative study.

PubMed

Timms, Mark; Steel, Rohan; Vine, John

2016-02-01

The recombinant human erythropoietins epoetin alfa (Eprex®), darbepoetin (Aranesp®) and methoxy polyethylene glycol-epoetin beta (Mircera®) were administered to greyhounds for 7, 10 and 14 days respectively. Blood and urine samples were collected and analysed for erythropoietin by ELISA, LC-MS/MS and western blotting. Limits of confirmation in plasma for western blotting and LC-MS/MS methods ranged from a low of 2.5mIU/mL, and closely matched the sensitivity of ELISA screening. Copyright © 2015 John Wiley & Sons, Ltd.

Validation of endothelin B receptor antibodies reveals two distinct receptor-related bands on Western blot.

PubMed

Barr, Travis P; Kornberg, Daniel; Montmayeur, Jean-Pierre; Long, Melinda; Reichheld, Stephen; Strichartz, Gary R

2015-01-01

Antibodies are important tools for the study of protein expression but are often used without full validation. In this study, we used Western blots to characterize antibodies targeted to the N or C terminal (NT or CT, respectively) and the second or third intracellular loop (IL2 or IL3, respectively) of the endothelin B receptor (ETB). The IL2-targeted antibody accurately detected endogenous ETB expression in rat brain and cultured rat astrocytes by labeling a 50-kDa band, the expected weight of full-length ETB. However, this antibody failed to detect transfected ETB in HEK293 cultures. In contrast, the NT-targeted antibody accurately detected endogenous ETB in rat astrocyte cultures and transfected ETB in HEK293 cultures by labeling a 37-kDa band but failed to detect endogenous ETB in rat brain. Bands detected by the CT- or IL3-targeted antibody were found to be unrelated to ETB. Our findings show that functional ETB can be detected at 50 or 37kDa on Western blot, with drastic differences in antibody affinity for these bands. The 37-kDa band likely reflects ETB processing, which appears to be dependent on cell type and/or culture condition. Copyright © 2014 Elsevier Inc. All rights reserved.

Immunodiagnosis of Echinococcus Infections: Confirmatory Testing and Species Differentiation by a New Commercial Western Blot

PubMed Central

Liance, Martine; Janin, Veronique; Bresson-Hadni, Solange; Vuitton, Dominique-Angele; Houin, Rene; Piarroux, Renaud

2000-01-01

The Echinococcus Western Blot IgG (LDBIO Diagnostics, Lyon, France), using a whole larval antigen from Echinococcus multilocularis, was evaluated for serodiagnosis and differentiation between two human parasitic infections of worldwide importance: cystic echinococcosis, due to Echinococcus granulosus, and alveolar echinococcosis, due to E. multilocularis. Fifty and 61 serum samples from patients with cystic and alveolar echinococcosis, respectively, were used for assessing diagnostic sensitivity. The sensitivity of the assay was compared with those of screening tests used for these applications. Sera used for assessing cross-reactivities were from 154 patients with other diseases, either parasitic or not. The assay allowed the detection of serum immunoglobulin G antibodies in 97% of Echinococcus-infected patients. It had a higher sensitivity than screening assays for the detection for each echinococcosis. The assay allowed us to correctly distinguish between E. granulosus- and E. multilocularis-infected patients in 76% of cases. It did not allow us to distinguish active from inactive forms of both echinococcoses. The occurrence of cross-reactivities with neurocysticercosis indicates the necessity for retesting sera with species-specific antigens, for rare patients with neurologic disorders. This study shows the usefulness of the commercially available Echinococcus Western Blot IgG for the serological confirmation of human echinococcosis. PMID:11015390

Southern Pine Beetle

Treesearch

Robert C. Thatcher; Patrick J. Barry

1982-01-01

The southern pine beetle (Dendroctonus frontalis Zimmermann) is one of pine's most destructive insect enemies in the Southern United States, Mexico, and Central America. Because populations build rapidly to outbreak proportions and large numbers of trees are killed, this insect generates considerable concern among managers of southern pine forests. The beetle...

A single-step simultaneous protein staining procedure for polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis.

PubMed

Pal, Jayanta K; Berwal, Sunil K; Soni, Rupali N

2012-01-01

A simple method for staining of proteins simultaneously on sodium dodecyl sulfate (SDS) polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis is described. A 5% solution of Alta, a commercially available cosmetic preparation, is added in the upper tank buffer during electrophoresis. On completion of electrophoresis, the gel is washed in distilled water and viewed on a white light plate and a transilluminator to photograph the protein profiles. The gel is processed for western blot transfer of proteins onto a nitrocellulose membrane, and upon completion, the protein profiles on the membrane are viewed and photographed as stated above. The membrane can then be processed for immunostaining as per the standard procedure. Thus, the staining procedure using Alta is simple, rapid (without any need of destaining), and cost-effective.

Anti-Peptide Monoclonal Antibodies Generated for Immuno-Multiple Reaction Monitoring-Mass Spectrometry Assays Have a High Probability of Supporting Western blot and ELISA*

PubMed Central

Schoenherr, Regine M.; Saul, Richard G.; Whiteaker, Jeffrey R.; Yan, Ping; Whiteley, Gordon R.; Paulovich, Amanda G.

2015-01-01

Immunoaffinity enrichment of peptides coupled to targeted, multiple reaction monitoring-mass spectrometry (immuno-MRM) has recently been developed for quantitative analysis of peptide and protein expression. As part of this technology, antibodies are generated to short, linear, tryptic peptides that are well-suited for detection by mass spectrometry. Despite its favorable analytical performance, a major obstacle to widespread adoption of immuno-MRM is a lack of validated affinity reagents because commercial antibody suppliers are reluctant to commit resources to producing anti-peptide antibodies for immuno-MRM while the market is much larger for conventional technologies, especially Western blotting and ELISA. Part of this reluctance has been the concern that affinity reagents generated to short, linear, tryptic peptide sequences may not perform well in traditional assays that detect full-length proteins. In this study, we test the feasibility and success rates of generating immuno-MRM monoclonal antibodies (mAbs) (targeting tryptic peptide antigens) that are also compatible with conventional, protein-based immuno-affinity technologies. We generated 40 novel, peptide immuno-MRM assays and determined that the cross-over success rates for using immuno-MRM monoclonals for Western blotting is 58% and for ELISA is 43%, which compare favorably to cross-over success rates amongst conventional immunoassay technologies. These success rates could most likely be increased if conventional and immuno-MRM antigen design strategies were combined, and we suggest a workflow for such a comprehensive approach. Additionally, the 40 novel immuno-MRM assays underwent fit-for-purpose analytical validation, and all mAbs and assays have been made available as a resource to the community via the Clinical Proteomic Tumor Analysis Consortium's (CPTAC) Antibody (http://antibodies.cancer.gov) and Assay Portals (http://assays.cancer.gov), respectively. This study also represents the first

Spatiotemporal dynamics of the Southern California Asian citrus psyllid (Diaphorina citri) invasion.

PubMed

Bayles, Brett R; Thomas, Shyam M; Simmons, Gregory S; Grafton-Cardwell, Elizabeth E; Daugherty, Mathew P

2017-01-01

Biological invasions are governed by spatial processes that tend to be distributed in non-random ways across landscapes. Characterizing the spatial and temporal heterogeneities of the introduction, establishment, and spread of non-native insect species is a key aspect of effectively managing their geographic expansion. The Asian citrus psyllid (Diaphorina citri), a vector of the bacterium associated with huanglongbing (HLB), poses a serious threat to commercial and residential citrus trees. In 2008, D. citri first began expanding northward from Mexico into parts of Southern California. Using georeferenced D. citri occurrence data from 2008-2014, we sought to better understand the extent of the geographic expansion of this invasive vector species. Our objectives were to: 1) describe the spatial and temporal distribution of D. citri in Southern California, 2) identify the locations of statistically significant D. citri hotspots, and 3) quantify the dynamics of anisotropic spread. We found clear evidence that the spatial and temporal distribution of D. citri in Southern California is non-random. Further, we identified the existence of statistically significant hotspots of D. citri occurrence and described the anisotropic dispersion across the Southern California landscape. For example, the dominant hotspot surrounding Los Angeles showed rapid and strongly asymmetric spread to the south and east. Our study demonstrates the feasibility of quantitative invasive insect risk assessment with the application of a spatial epidemiology framework.

Spatiotemporal dynamics of the Southern California Asian citrus psyllid (Diaphorina citri) invasion

PubMed Central

Thomas, Shyam M.; Simmons, Gregory S.; Grafton-Cardwell, Elizabeth E.; Daugherty, Mathew P.

2017-01-01

Biological invasions are governed by spatial processes that tend to be distributed in non-random ways across landscapes. Characterizing the spatial and temporal heterogeneities of the introduction, establishment, and spread of non-native insect species is a key aspect of effectively managing their geographic expansion. The Asian citrus psyllid (Diaphorina citri), a vector of the bacterium associated with huanglongbing (HLB), poses a serious threat to commercial and residential citrus trees. In 2008, D. citri first began expanding northward from Mexico into parts of Southern California. Using georeferenced D. citri occurrence data from 2008–2014, we sought to better understand the extent of the geographic expansion of this invasive vector species. Our objectives were to: 1) describe the spatial and temporal distribution of D. citri in Southern California, 2) identify the locations of statistically significant D. citri hotspots, and 3) quantify the dynamics of anisotropic spread. We found clear evidence that the spatial and temporal distribution of D. citri in Southern California is non-random. Further, we identified the existence of statistically significant hotspots of D. citri occurrence and described the anisotropic dispersion across the Southern California landscape. For example, the dominant hotspot surrounding Los Angeles showed rapid and strongly asymmetric spread to the south and east. Our study demonstrates the feasibility of quantitative invasive insect risk assessment with the application of a spatial epidemiology framework. PMID:28278188

Single-Cell Western Blotting after Whole-Cell Imaging to Assess Cancer Chemotherapeutic Response

PubMed Central

2015-01-01

Intratumor heterogeneity remains a major obstacle to effective cancer therapy and personalized medicine. Current understanding points to differential therapeutic response among subpopulations of tumor cells as a key challenge to successful treatment. To advance our understanding of how this heterogeneity is reflected in cell-to-cell variations in chemosensitivity and expression of drug-resistance proteins, we optimize and apply a new targeted proteomics modality, single-cell western blotting (scWestern), to a human glioblastoma cell line. To acquire both phenotypic and proteomic data on the same, single glioblastoma cells, we integrate high-content imaging prior to the scWestern assays. The scWestern technique supports thousands of concurrent single-cell western blots, with each assay comprised of chemical lysis of single cells seated in microwells, protein electrophoresis from those microwells into a supporting polyacrylamide (PA) gel layer, and in-gel antibody probing. We systematically optimize chemical lysis and subsequent polyacrylamide gel electrophoresis (PAGE) of the single-cell lysate. The scWestern slides are stored for months then reprobed, thus allowing archiving and later analysis as relevant to sparingly limited, longitudinal cell specimens. Imaging and scWestern analysis of single glioblastoma cells dosed with the chemotherapeutic daunomycin showed both apoptotic (cleaved caspase 8- and annexin V-positive) and living cells. Intriguingly, living glioblastoma subpopulations show up-regulation of a multidrug resistant protein, P-glycoprotein (P-gp), suggesting an active drug efflux pump as a potential mechanism of drug resistance. Accordingly, linking of phenotype with targeted protein analysis with single-cell resolution may advance our understanding of drug response in inherently heterogeneous cell populations, such as those anticipated in tumors. PMID:25226230

Current subsidence rates due to compaction of Holocene sediments in southern Louisiana

USGS Publications Warehouse

Meckel, T.A.; ten Brink, Uri S.; Williams, S.J.

2006-01-01

Relative contributions of geologic and anthropogenic processes to subsidence of southern Louisiana are vigorously debated. Of these, shallow sediment compaction is often considered dominant, although this has never been directly observed or effectively demonstrated. Quantitative understanding of subsidence is important for predicting relative sea level rise, storm surge flooding due to hurricanes, and for successful wetland restoration. Despite many shallow borings, few appropriate stratigraphic and geotechnical data are available for site-specific calculations. We overcome this by determining present compaction rates from Monte Carlo simulations of the incremental sedimentation and compaction of stratigraphies typical of the Holocene of southern Louisiana. This approach generates distributions of present compaction rates that are not expected to exceed 5 mm/yr, but may locally. Locations with present subsidence rates greater than the predicted maximum probable shallow compaction rates are likely influenced by additional processes.

Outbreak of larval Echinococcus multilocularis infection in Japanese monkey (Macaca fuscata) in a zoo, Hokkaido: western blotting patterns in the infected monkeys.

PubMed

Sato, Chiaki; Kawase, Shiro; Yano, Shoki; Nagano, Hideki; Fujimoto, Satoshi; Kobayashi, Nobuyuki; Miyahara, Kazuro; Yamada, Kazutaka; Sato, Motoyoshi; Kobayashi, Yoshiyasu

2005-01-01

A high prevalence of larval Echinococcus multilocularis (Em) infection was found in zoo primates in Hokkaido, Japan. In October 1997, a Japanese monkey (Macaca fuscata) died and histopathologically diagnosed as alveolar hydatidosis. Serum samples were collected from the remaining Japanese monkeys and examined for antibodies against Em by enzyme-linked immunosorbent assay and western blotting. Serological tests showed 12 more animals of the remaining 57 monkeys were possibly infected. Ultrasonography revealed that nine of these 12 animals had a cystic lesion in the liver. The band patterns of western blotting in the monkeys were very similar to those in human.

Proviral Features of Human T Cell Leukemia Virus Type 1 in Carriers with Indeterminate Western Blot Analysis Results.

PubMed

Kuramitsu, Madoka; Sekizuka, Tsuyoshi; Yamochi, Tadanori; Firouzi, Sanaz; Sato, Tomoo; Umeki, Kazumi; Sasaki, Daisuke; Hasegawa, Hiroo; Kubota, Ryuji; Sobata, Rieko; Matsumoto, Chieko; Kaneko, Noriaki; Momose, Haruka; Araki, Kumiko; Saito, Masumichi; Nosaka, Kisato; Utsunomiya, Atae; Koh, Ki-Ryang; Ogata, Masao; Uchimaru, Kaoru; Iwanaga, Masako; Sagara, Yasuko; Yamano, Yoshihisa; Okayama, Akihiko; Miura, Kiyonori; Satake, Masahiro; Saito, Shigeru; Itabashi, Kazuo; Yamaguchi, Kazunari; Kuroda, Makoto; Watanabe, Toshiki; Okuma, Kazu; Hamaguchi, Isao

2017-09-01

Western blotting (WB) for human T cell leukemia virus type 1 (HTLV-1) is performed to confirm anti-HTLV-1 antibodies detected at the initial screening of blood donors and in pregnant women. However, the frequent occurrence of indeterminate results is a problem with this test. We therefore assessed the cause of indeterminate WB results by analyzing HTLV-1 provirus genomic sequences. A quantitative PCR assay measuring HTLV-1 provirus in WB-indeterminate samples revealed that the median proviral load was approximately 100-fold lower than that of WB-positive samples (0.01 versus 0.71 copy/100 cells). Phylogenic analysis of the complete HTLV-1 genomes of WB-indeterminate samples did not identify any specific phylogenetic groups. When we analyzed the nucleotide changes in 19 HTLV-1 isolates from WB-indeterminate samples, we identified 135 single nucleotide substitutions, composed of four types, G to A (29%), C to T (19%), T to C (19%), and A to G (16%). In the most frequent G-to-A substitution, 64% occurred at GG dinucleotides, indicating that APOBEC3G is responsible for mutagenesis in WB-indeterminate samples. Moreover, interestingly, five WB-indeterminate isolates had nonsense mutations in Pol and/or Tax, Env, p12, and p30. These findings suggest that WB-indeterminate carriers have low production of viral antigens because of a combination of a low proviral load and mutations in the provirus, which may interfere with host recognition of HTLV-1 antigens. Copyright © 2017 American Society for Microbiology.

Integral Phylogenomic Approach over Ilex L. Species from Southern South America

PubMed Central

Cascales, Jimena; Bracco, Mariana; Garberoglio, Mariana J.; Poggio, Lidia; Gottlieb, Alexandra M.

2017-01-01

The use of molecular markers with inadequate variation levels has resulted in poorly resolved phylogenetic relationships within Ilex. Focusing on southern South American and Asian species, we aimed at contributing informative plastid markers. Also, we intended to gain insights into the nature of morphological and physiological characters used to identify species. We obtained the chloroplast genomes of I. paraguariensis and I. dumosa, and combined these with all the congeneric plastomes currently available to accomplish interspecific comparisons and multilocus analyses. We selected seven introns and nine IGSs as variable non-coding markers that were used in phylogenomic analyses. Eight extra IGSs were proposed as candidate markers. Southern South American species formed one lineage, except for I. paraguariensis, I. dumosa and I. argentina, which occupied intermediate positions among sampled taxa; Euroasiatic species formed two lineages. Some concordant relationships were retrieved from nuclear sequence data. We also conducted integral analyses, involving a supernetwork of molecular data, and a simultaneous analysis of quantitative and qualitative morphological and phytochemical characters, together with molecular data. The total evidence tree was used to study the evolution of non-molecular data, evidencing fifteen non-ambiguous synapomorphic character states and consolidating the relationships among southern South American species. More South American representatives should be incorporated to elucidate their origin. PMID:29165335

Southern Living and Southern Voices: Models of Regional Magazine Success and Failure.

ERIC Educational Resources Information Center

English, John W.

This paper examines the phenomenon of magazine success and failure as demonstrated by two regional magazines, "Southern Living" and "Southern Voices." The former, a combination of articles about food, travel, sports, and other positive aspects of southern life, was quickly accepted by its readers and advertisers and began…

PCR colorimetric dot-blot assay and clinical pretest probability for diagnosis of Pulmonary Tuberculosis in Smear-Negative patients

PubMed Central

Scherer, Luciene Cardoso; Sperhacke, Rosa Dea; Jarczewski, Carla; Cafrune, Patrícia I; Minghelli, Simone; Ribeiro, Marta Osório; Mello, Fernanda CQ; Ruffino-Netto, Antonio; Rossetti, Maria LR; Kritski, Afrânio L

2007-01-01

Background Smear-negative pulmonary tuberculosis (SNPTB) accounts for 30% of Pulmonary Tuberculosis (PTB) cases reported annually in developing nations. Polymerase chain reaction (PCR) may provide an alternative for the rapid detection of Mycobacterium tuberculosis (MTB); however little data are available regarding the clinical utility of PCR in SNPTB, in a setting with a high burden of TB/HIV co-infection. Methods To evaluate the performance of the PCR dot-blot in parallel with pretest probability (Clinical Suspicion) in patients suspected of having SNPTB, a prospective study of 213 individuals with clinical and radiological suspicion of SNPTB was carried out from May 2003 to May 2004, in a TB/HIV reference hospital. Respiratory specialists estimated the pretest probability of active disease into high, intermediate, low categories. Expectorated sputum was examined by direct microscopy (Ziehl-Neelsen staining), culture (Lowenstein Jensen) and PCR dot-blot. Gold standard was based on culture positivity combined with the clinical definition of PTB. Results In smear-negative and HIV subjects, active PTB was diagnosed in 28.4% (43/151) and 42.2% (19/45), respectively. In the high, intermediate and low pretest probability categories active PTB was diagnosed in 67.4% (31/46), 24% (6/25), 7.5% (6/80), respectively. PCR had sensitivity of 65% (CI 95%: 50%–78%) and specificity of 83% (CI 95%: 75%–89%). There was no difference in the sensitivity of PCR in relation to HIV status. PCR sensitivity and specificity among non-previously TB treated and those treated in the past were, respectively: 69%, 43%, 85% and 80%. The high pretest probability, when used as a diagnostic test, had sensitivity of 72% (CI 95%:57%–84%) and specificity of 86% (CI 95%:78%–92%). Using the PCR dot-blot in parallel with high pretest probability as a diagnostic test, sensitivity, specificity, positive and negative predictive values were: 90%, 71%, 75%, and 88%, respectively. Among non

Anti-RAINBOW dye-specific antibodies as universal tools for the visualization of prestained protein molecular weight markers in Western blot analysis.

PubMed

Schüchner, Stefan; Andorfer, Peter; Mudrak, Ingrid; Ogris, Egon

2016-08-17

Western blotting is one of the most widely used techniques in molecular biology and biochemistry. Prestained proteins are used as molecular weight standards in protein electrophoresis. In the chemiluminescent Western blot analysis, however, these colored protein markers are invisible leaving researchers with the unsatisfying situation that the signal for the protein of interest and the signal for the markers are not captured simultaneously and have to be merged in an error-prone step. To allow the simultaneous detection of marker proteins we generated monoclonal antibodies specific for the protein dyes. To elicit a dye rather than protein specific immune response we immunized mice sequentially with dye-carrier protein complexes, in which a new carrier protein was used for each subsequent immunization. Moreover, by sequentially immunizing with dye-carrier protein complexes, in which different but structurally related dyes were used, we could also generate an antibody, termed anti-RAINBOW, that cross-reacted even with structurally related dyes not used in the immunizations. Our novel antibodies represent convenient tools for the simultaneous Western blot detection of commercially available prestained marker proteins in combination with the detection of any specific protein of interest. These antibodies will render obsolete the anachronistic tradition of manually charting marker bands on film.

Anti-RAINBOW dye-specific antibodies as universal tools for the visualization of prestained protein molecular weight markers in Western blot analysis

PubMed Central

Schüchner, Stefan; Andorfer, Peter; Mudrak, Ingrid; Ogris, Egon

2016-01-01

Western blotting is one of the most widely used techniques in molecular biology and biochemistry. Prestained proteins are used as molecular weight standards in protein electrophoresis. In the chemiluminescent Western blot analysis, however, these colored protein markers are invisible leaving researchers with the unsatisfying situation that the signal for the protein of interest and the signal for the markers are not captured simultaneously and have to be merged in an error-prone step. To allow the simultaneous detection of marker proteins we generated monoclonal antibodies specific for the protein dyes. To elicit a dye rather than protein specific immune response we immunized mice sequentially with dye-carrier protein complexes, in which a new carrier protein was used for each subsequent immunization. Moreover, by sequentially immunizing with dye-carrier protein complexes, in which different but structurally related dyes were used, we could also generate an antibody, termed anti-RAINBOW, that cross-reacted even with structurally related dyes not used in the immunizations. Our novel antibodies represent convenient tools for the simultaneous Western blot detection of commercially available prestained marker proteins in combination with the detection of any specific protein of interest. These antibodies will render obsolete the anachronistic tradition of manually charting marker bands on film. PMID:27531616

Meteorological Controls on Biomass Burning During Santa Ana Events in Southern California

NASA Technical Reports Server (NTRS)

Veraverbeke, Sander; Capps, Scott; Hook, Simon J.; Randerson, James T.; Jin, Yufang; Hall, Alex

2013-01-01

Fires occurring during Santa Ana (SA) events in southern California are driven by extreme fire weather characterized by high temperatures, low humidities, and high wind speeds. We studied the controls on burned area and carbon emissions during two intensive SA burning periods in 2003 and 2007. We therefore used remote sensing data in parallel with fire weather simulations of the Weather and Regional Forecast model. Total carbon emissions were approximately 1800 gigagrams in 2003 and 900 gigagrams in 2007, based on a daily burned area and a fire emission model that accounted for spatial variability in fuel loads and combustion completeness. On a regional scale, relatively strong positive correlations were found between the daily Fosberg fire weather index and burned area/emissions (probability is less than 0.01). Our analysis provides a quantitative assessment of relationships between fire activity and weather during severe SA fires in southern California.

Development of EMab-51, a Sensitive and Specific Anti-Epidermal Growth Factor Receptor Monoclonal Antibody in Flow Cytometry, Western Blot, and Immunohistochemistry.

PubMed

Itai, Shunsuke; Kaneko, Mika K; Fujii, Yuki; Yamada, Shinji; Nakamura, Takuro; Yanaka, Miyuki; Saidoh, Noriko; Handa, Saori; Chang, Yao-Wen; Suzuki, Hiroyoshi; Harada, Hiroyuki; Kato, Yukinari

2017-10-01

The epidermal growth factor receptor (EGFR) is a member of the human epidermal growth factor receptor (HER) family of receptor tyrosine kinases and is involved in cell growth and differentiation. EGFR homodimers or heterodimers with other HER members, such as HER2 and HER3, activate downstream signaling cascades in many cancers. In this study, we developed novel anti-EGFR monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. First, we expressed the full-length or ectodomain of EGFR in LN229 glioblastoma cells and then immunized mice with LN229/EGFR or ectodomain of EGFR, and performed the first screening using enzyme-linked immunosorbent assays. Subsequently, we selected mAbs according to their efficacy in flow cytometry (second screening), Western blot (third screening), and immunohistochemical (fourth screening) analyses. Among 100 mAbs, only one clone EMab-51 (IgG 1 , kappa) reacted with EGFR in Western blot analysis. Finally, immunohistochemical analyses with EMab-51 showed sensitive and specific reactions against oral cancer cells, warranting the use of EMab-51 to detect EGFR in pathological analyses of EGFR-expressing cancers.

A Quantitative Study on Burnout for Teachers Who Work with Students Who Have Moderate to Severe Disabilities

ERIC Educational Resources Information Center

Dickerson, Elizabeth G.

2017-01-01

Purpose: The purpose of this quantitative research was to examine what relationships, if any, exist between the independent variable of burnout and dependent variables of job satisfaction for special education teachers who work with students who have moderate to severe disabilities ages 5 to 22 in a Southern California school district.…

An advanced dual labeled gold nanoparticles probe to detect Cryptosporidium parvum using rapid immuno-dot blot assay.

PubMed

Thiruppathiraja, Chinnasamy; Kamatchiammal, Senthilkumar; Adaikkappan, Periyakaruppan; Alagar, Muthukaruppan

2011-07-15

The zoonotic protozoan parasite Cryptosporidium parvum poses a significant risk to public health. Due to the low infectious dose of C. parvum, remarkably sensitive detection methods are required for water and food industries analysis. However PCR affirmed sensing method of the causative nucleic acid has numerous advantages, still criterion demands for simple techniques and expertise understanding to extinguish its routine use. In contrast, protein based immuno detecting techniques are simpler to perform by a commoner, but lack of sensitivity due to inadequate signal amplification. In this paper, we focused on the development of a mere sensitive immuno detection method by coupling anti-cyst antibody and alkaline phosphatase on gold nanoparticle for C. parvum is described. Outcome of the sensitivity in an immuno-dot blot assay detection is enhanced by 500 fold (using conventional method) and visually be able to detect up to 10 oocysts/mL with minimal processing period. Techniques reported in this paper substantiate the convenience of immuno-dot blot assay for the routine screening of C. parvum in water/environmental examines and most importantly, demonstrates the potential of a prototype development of simple and inexpensive diagnostic technique. Copyright © 2011 Elsevier B.V. All rights reserved.

Cy5 total protein normalization in Western blot analysis.

PubMed

Hagner-McWhirter, Åsa; Laurin, Ylva; Larsson, Anita; Bjerneld, Erik J; Rönn, Ola

2015-10-01

Western blotting is a widely used method for analyzing specific target proteins in complex protein samples. Housekeeping proteins are often used for normalization to correct for uneven sample loads, but these require careful validation since expression levels may vary with cell type and treatment. We present a new, more reliable method for normalization using Cy5-prelabeled total protein as a loading control. We used a prelabeling protocol based on Cy5 N-hydroxysuccinimide ester labeling that produces a linear signal response. We obtained a low coefficient of variation (CV) of 7% between the ratio of extracellular signal-regulated kinase (ERK1/2) target to Cy5 total protein control signals over the whole loading range from 2.5 to 20.0μg of Chinese hamster ovary cell lysate protein. Corresponding experiments using actin or tubulin as controls for normalization resulted in CVs of 13 and 18%, respectively. Glyceraldehyde-3-phosphate dehydrogenase did not produce a proportional signal and was not suitable for normalization in these cells. A comparison of ERK1/2 signals from labeled and unlabeled samples showed that Cy5 prelabeling did not affect antibody binding. By using total protein normalization we analyzed PP2A and Smad2/3 levels with high confidence. Copyright © 2015 Elsevier Inc. All rights reserved.

Glacial-Interglacial Variability of Nd isotopes in the South Atlantic and Southern Ocean

NASA Astrophysics Data System (ADS)

Knudson, K. P.; Goldstein, S. L.; Pena, L.; Seguí, M. J.; Kim, J.; Yehudai, M.; Fahey, T.

2017-12-01

Understanding the relationship between meridional overturning circulation and climate is key to understanding the processes and feedbacks underlying future climate changes. North Atlantic Deep Water (NADW) represents a major water mass that participates in global oceanic circulation and undergoes substantial reorganization with climate changes on millennial and orbital timescales. Nd isotopes are semi-quantitative water mass tracers that reflect the mixing of end-member water masses, and their values in the Southern Ocean offer the ability to characterize NADW variability over time. Here, we present paleo-circulation records of Nd isotopes measured on fish debris and Fe-Mn encrusted foraminifera from ODP Sites 1090 (42° 54.82'S, 3702 m), and 1094 (53° 10.81'S, 2807 m). Site 1090 is located in the Cape Basin, SE Atlantic, near the lower boundary between NADW and Circumpolar Deep Water (CDW), while 1094 is in the Circumpolar Current. They are ideal locations to monitor changes in the export of NADW to the Southern Ocean. These new results build on previous work (Pena and Goldstein, 2014) to document meridional overturning changes in the Southern Ocean.

Diatom species abundance and morphologically-based dissolution proxies in coastal Southern Ocean assemblages

NASA Astrophysics Data System (ADS)

Warnock, Jonathan P.; Scherer, Reed P.

2015-07-01

Taphonomic processes alter diatom assemblages in sediments, thus potentially negatively impacting paleoclimate records at various rates across space, time, and taxa. However, quantitative taphonomic data is rarely included in diatom-based paleoenvironmental reconstructions and no objective standard exists for comparing diatom dissolution in sediments recovered from marine depositional settings, including the Southern Ocean's opal belt. Furthermore, identifying changes to diatom dissolution through time can provide insight into the efficiency of both upper water column nutrient recycling and the biological pump. This is significant in that reactive metal proxies (e.g. Al, Ti) in the sediments only account for post-depositional dissolution, not the water column where the majority of dissolution occurs. In order to assess the range of variability of responses to dissolution in a typical Southern Ocean diatom community and provide a quantitative guideline for assessing taphonomic variability in diatoms recovered from core material, a sediment trap sample was subjected to controlled, serial dissolution. By evaluating dissolution-induced changes to diatom species' relative abundance, three preservational categories of diatoms have been identified: gracile, intermediate, and robust. The relative abundances of these categories can be used to establish a preservation grade for diatom assemblages. However, changes to the relative abundances of diatom species in sediment samples may reflect taphonomic or ecological factors. In order to address this complication, relative abundance changes have been tied to dissolution-induced morphological change to the areolae of Fragilariopsis curta, a significant sea-ice indicator in Southern Ocean sediments. This correlation allows differentiation between gracile species loss to dissolution versus ecological factors or sediment winnowing. These results mirror a similar morphological dissolution index from a parallel study utilizing

Application of a Dot Blot Hybridization Platform to Assess Streptococcus uberis Population Structure in Dairy Herds

PubMed Central

Albuquerque, Pedro; Ribeiro, Niza; Almeida, Alexandre; Panschin, Irena; Porfirio, Afonso; Vales, Marta; Diniz, Francisca; Madeira, Helena; Tavares, Fernando

2017-01-01

Streptococcus uberis is considered one of the most important pathogens associated with bovine mastitis. While traditionally acknowledged as an environmental pathogen, S. uberis has been shown to adopt a contagious epidemiological pattern in several dairy herds. Since different control strategies are employed depending on the mode of transmission, in-depth studies of S. uberis populations are essential to determine the best practices to control this pathogen. In this work, we optimized and validated a dot blot platform, combined with automatic image analysis, to rapidly assess the population structure of infective S. uberis, and evaluated its efficiency when compared to multilocus sequence analysis (MLSA) genotyping. Two dairy herds with prevalent S. uberis infections were followed in a 6 month period, in order to collect and characterize isolates from cows with persistent infections. These herds, located in Portugal (Barcelos and Maia regions), had similar management practices, with the herd from Barcelos being smaller and having a better milking parlor management, since infected cow segregation was immediate. A total of 54 S. uberis isolates were obtained from 24 different cows from the two herds. To overcome operator-dependent analysis of the dot blots and increase the technique's consistency and reliability, the hybridization signals were converted into probability values, with average probabilities higher than 0.5 being considered positive results. These data allowed to confirm the isolates' identity as S. uberis using taxa-specific markers and to determine the presence of virulence- and antibiotic resistance-related genes. In addition, MLSA allowed to disclose the most prevalent S. uberis clonal lineages in both herds. Seven different clusters were identified, with Barcelos showing a high clonal diversity and Maia a dominant lineage infecting most cows, suggesting distinct epidemiological patterns, with S. uberis displaying an environmental or contagious

Leadership Style of Head Teachers of Basic Special Schools as Correlates of Retention of Special Needs Educators in Southern Ghana

ERIC Educational Resources Information Center

Kumedzro, Felix Kwame; Otube, Nelly; Wamunyi, Chomba; Runo, Mary

2016-01-01

The study aimed at establishing relationship between leadership style of head teachers and retention of special education teachers in Southern Ghana. The study was purely quantitative and utilized descriptive correlation design which allowed the researcher to establish the strength and direction of the relationship between the independent variable…

Monitoring Across Borders: 2010 Joint Meeting of the Forest Inventory and Analysis (FIA) Symposium and the Southern Mensurationists

Treesearch

Will McWilliams; Francis A. Roesch

2012-01-01

These proceedings represent the range of topics covered during the 2010 Joint Meeting of the Forest Inventory and Analysis (FIA) Symposium and the Southern Mensurationists, October 5-7, 2010 in Knoxville, TN. The meeting was a gathering of forest scientists with a quantitative leaning and, as such, the papers discuss the aspects of the observation, estimation, modeling...

An assessment of a Western blot method for the investigation of canine cutaneous adverse food reactions.

PubMed

Maina, Elisa; Matricoti, Irina; Noli, Chiara

2018-06-01

Adverse food reaction (AFR) is diagnosed with a two month elimination diet (ED), followed by challenge with the original food. To evaluate reactivity of selected EDs and performance of a Western blot serological test for the diagnosis of AFR. Twenty five food reactive (FR) and 13 non food reactive (NFR) privately owned dogs. Sera were tested for antibodies against hydrolyzed poultry feather (RCA), hydrolyzed soy (PHA), hydrolyzed fish (FUH), limited antigen horse and potato (THP), fresh horse meat and the offending food for each FR dog as documented by provocative challenge. Fourteen sera were negative and two positive to all foods. Sera from five of 13 NFR and three of 25 FR dogs were reactive to hydrolyzed foods. The RCA diet was recognized by four of 38, FUH by six of 38 and PHA by one of 28 samples. THP was recognized by 14 of 33 and fresh horse by one of ten dogs that had never eaten horse meat. The test correctly identified one of 15 dogs allergic to FUH. Twenty of 25 FR sera were negative for the dogs' respective offending foods (20% sensitivity), whereas four of 13 NFR sera were positive to the dogs' usual diets (69% specificity). Western blot analysis cannot be considered as a valid tool for the diagnosis of AFR; it may serve as an aid in selecting an ED. © 2018 ESVD and ACVD.

Vegetation history and palaeoclimate of the last glacial period at Lago Grande di Monticchio, Southern Italy

NASA Astrophysics Data System (ADS)

Watts, W. A.; Allen, J. R. M.; Huntley, B.

A high-resolution palynological study of a 51 m core from Lago Grande di Monticchio, southern Italy, has provided a palaeonvironmental record for the last glacill. A annual lamination based chronology, supported by radiometric and tephrochronological dates, provides an absolute timescale for this record that spans 76,300 years. Correlations are established between the pollen stratigraphy, the GRIP ice core δ18O record and foraminiferal assemblages from Atlantic core V23-81. Both Dansgaard-Oeschger and Heinrich events are reflected by changes in the pollen stratigraphy. Revised dates are estimated for Heinrich events H1-H6. A quantitative palaeoclimate reconstruction based upon the pollen data provides evidence of the climate changes in southern Italy associated with these and other fluctuations during the last glacial.

A collaborative approach to advance student research at the University of Southern California.

PubMed

Stephens, Heather; Jensen, Bridger; Carpiaux, Weston; Sedghizadeh, Parish; Chai, Yang

2012-05-01

The continued advancement of oral health and science relies upon the cultivation of a student's interest in research. The Student Research Group at the Ostrow School of Dentistry of the University of Southern California is working to increase student involvement in research and develop future academic leaders. This study aims to, through student surveys, quantitatively evaluate students' involvement in research, students' interest in participating in research and to identify specific barriers students feel challenge their ability to participate in research.

Detection and quantitation of HER-2/neu gene amplification in human breast cancer archival material using fluorescence in situ hybridization.

PubMed

Pauletti, G; Godolphin, W; Press, M F; Slamon, D J

1996-07-04

Amplification and overexpression of the HER-2/neu gene occurs in 25-30% of human breast cancers. This genetic alteration is associated with a poor clinical prognosis in women with either node negative or node positive breast cancers. The initial studies testing this association were somewhat controversial and this controversy was due in large part to significant heterogeneity in both the methods and/or reagents used in testing archival material for the presence of the alteration. These methods included a number of solid matrix blotting techniques for DNA, RNA and protein as well as immunohistochemistry. Fluorescence in situ hybridization (FISH) represents the newest methodologic approach for testing for this genetic alteration. In this study, FISH is compared to Southern, Northern and Western blot analyses as well as immunohistochemistry in a large cohort of archival human breast cancer specimens. FISH was found to be superior to all other methodologies tested in assessing formalin fixed, paraffin embedded material for HER-2/neu amplification. The results from this study also confirm that overexpression of HER-2/neu rarely occurs in the absence of gene amplification in breast cancer (approximately 3% of cases). This method of analysis is rapid, reproducible and extremely reliable in detecting presence of HER-2/neu gene amplification and should have clinical utility.

76 FR 35508 - Alabama Southern Railroad, L.L.C.-Temporary Trackage Rights Exemption-Norfolk Southern Railway...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-17

... DEPARTMENT OF TRANSPORTATION Surface Transportation Board [Docket No. FD 35510] Alabama Southern Railroad, L.L.C.--Temporary Trackage Rights Exemption--Norfolk Southern Railway Company Norfolk Southern... grant nonexclusive overhead temporary trackage rights to Alabama Southern Railroad, L.L.C. (ABS) over a...

Southern Pine Beetle Information System (SPBIS)

Treesearch

Valli Peacher

2011-01-01

The southern pine beetle (SPB) is the most destructive forest insect in the South. The SPB attacks all species of southern pine, but loblolly and shortleaf are most susceptible. The Southern Pine Beetle Information System (SPBIS) is the computerized database used by the national forests in the Southern Region for tracking individual southern pine beetle infestations....

Southern Pine Beetle Competitors

Treesearch

Fred M. Stephen

2011-01-01

When southern pine beetles mass attack a living pine tree, if colonization is successful the tree dies and its phloem becomes immediately available to a complex of other bark beetles and long-horned beetles, all of which, in order to reproduce, compete for the new resource. In southern pines the phloem-inhabiting guild is composed of the southern pine beetle...

Contribution of a Comparative Western Blot Method to Early Postnatal Diagnosis of Congenital Syphilis.

PubMed

Marangoni, Antonella; Foschi, Claudio; Capretti, Maria Grazia; Nardini, Paola; Compri, Monica; Corvaglia, Luigi Tommaso; Faldella, Giacomo; Cevenini, Roberto

2016-05-01

Serology has a pivotal role in the diagnosis of congenital syphilis (CS), but problems arise because of the passive transfer of IgG antibodies across the placenta. The aim of this study was to assess the diagnostic value of a comparative Western blot (WB) method finalized to match the IgG immunological profiles of mothers and their own babies at birth in order to differentiate between passively transmitted maternal antibodies and antibodies synthesized by the infants against Treponema pallidum Thirty infants born to mothers with unknown or inadequate treatment for syphilis were entered in a retrospective study, conducted at St. Orsola-Malpighi Hospital, Bologna, Italy. All of the infants underwent clinical, instrumental, and laboratory examinations, including IgM WB testing. For the retrospective study, an IgG WB assay was performed by blotting T. pallidum antigens onto nitrocellulose sheets and incubating the strips with serum specimens from mother-child pairs. CS was diagnosed in 11 out of the 30 enrolled infants; 9/11 cases received the definitive diagnosis within the first week of life, whereas the remaining two were diagnosed later because of increasing serological test titers. The use of the comparative IgG WB testing performed with serum samples from mother-child pairs allowed a correct CS diagnosis in 10/11 cases. The CS diagnosis was improved by a strategy combining comparative IgG WB results with IgM WB results, leading to a sensitivity of 100%. The comparative IgG WB test is thus a welcome addition to the conventional laboratory methods used for CS diagnosis, allowing identification and adequate treatment of infected infants and avoiding unnecessary therapy of uninfected newborns. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

The effects of climate change on the demand for municipal water for residential landscaping in Southern Nevada

NASA Astrophysics Data System (ADS)

Tchigriaeva, E.; Lott, C.; Rollins, K.

2013-12-01

We analyze urban residential water demand for Southern Nevada as a part of the Nevada Infrastructure for Climate Change Science, Education, and Outreach project. The Nevada Climate Change project is a statewide interdisciplinary program which has launched joint research, education, and outreach on the effects of regional climate change on ecosystem services in Nevada with a particular focus on water resources. We estimate a random effect multiple regression model of urban residential water demand in order to better understand how residential water use is impacted by weather conditions and landscape characteristics and ultimately to inform predictions of urban water demand. The project develops a methodology of unification for several datasets from various sources including the Las Vegas Valley Water District (LVVWD), the Southern Nevada Water Authority (SNWA), Clark County Assessor, and the National Climatic Data Center (NCDC) resulting in a sample of 3,671,983 observations for 62,237 households with uninterrupted water use history for Las Vegas urban residents for the period from February 2007 to December 2011. The presented results (i) are significantly robust and in accordance with the economics theories, (ii) support basic empirical knowledge of weather and surface influence on water outdoor consumption, (iii) suggest quantitative measurements for predicting future water use due to climate/temperature changes as well as landscape redesign practices, and (iv) provide quantitative evaluation of the effectiveness of the existing water conservation programs by the Southern Nevada Water Authority (SNWA). The further study of conservation programs and analysis of interactions between surfaces and weather using the developed approach looks promising.

Analysis of Chromatin Organisation

ERIC Educational Resources Information Center

Szeberenyi, Jozsef

2011-01-01

Terms to be familiar with before you start to solve the test: chromatin, nucleases, sucrose density gradient centrifugation, melting point, gel electrophoresis, ethidium bromide, autoradiography, Southern blotting, Northern blotting, Sanger sequencing, restriction endonucleases, exonucleases, linker DNA, chloroform extraction, nucleosomes,…

77 FR 17122 - Indiana Southern Railroad, LLC-Temporary Trackage Rights Exemption-Norfolk Southern Railway Company

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-23

... Railroad, LLC--Temporary Trackage Rights Exemption--Norfolk Southern Railway Company Norfolk Southern Railway Company (NSR), pursuant to a written trackage rights agreement (Agreement), has agreed to grant overhead temporary trackage rights to Indiana Southern Railroad, LLC (ISRR) over NSR's line of railroad...

Fertilizing Southern Hardwoods

Treesearch

W. M. Broadfoot; A. F. Ike

1967-01-01

If present trends continue, fertilizing may soon be economically feasible in southern hardwood stands. Demands for the wood are rising, and the acreage alloted for growing it is steadily shrinking. To supply anticipated requests for information, the U. S. Forest Service has established tree nutrition studies at the Southern Hardwoods Laboratory in Stoneville,...

Development of a Sequence-Characterized Amplified Region Marker-Targeted Quantitative PCR Assay for Strain-Specific Detection of Oenococcus oeni during Wine Malolactic Fermentation▿

PubMed Central

Solieri, Lisa; Giudici, Paolo

2010-01-01

Control over malolactic fermentation (MLF) is a difficult goal in winemaking and needs rapid methods to monitor Oenococcus oeni malolactic starters (MLS) in a stressful environment such as wine. In this study, we describe a novel quantitative PCR (QPCR) assay enabling the detection of an O. oeni strain during MLF without culturing. O. oeni strain LB221 was used as a model to develop a strain-specific sequence-characterized amplified region (SCAR) marker derived from a discriminatory OPA20-based randomly amplified polymorphic DNA (RAPD) band. The 5′ and 3′ flanking regions and the copy number of the SCAR marker were characterized using inverse PCR and Southern blotting, respectively. Primer pairs targeting the SCAR sequence enabled strain-specific detection without cross amplification of other O. oeni strains or wine species of lactic acid bacteria (LAB), acetic acid bacteria (AAB), and yeasts. The SCAR-QPCR assay was linear over a range of cell concentrations (7 log units) and detected as few as 2.2 × 102 CFU per ml of red wine with good quantification effectiveness, as shown by the correlation of QPCR and plate counting results. Therefore, the cultivation-independent monitoring of a single O. oeni strain in wine based on a SCAR marker represents a rapid and effective strain-specific approach. This strategy can be adopted to develop easy and rapid detection techniques for monitoring the implantation of inoculated O. oeni MLS on the indigenous LAB population, reducing the risk of unsuccessful MLF. PMID:20935116

The antimicrobial resistance patterns and associated determinants in Streptococcus suis isolated from humans in southern Vietnam, 1997-2008

PubMed Central

2011-01-01

Background Streptococcus suis is an emerging zoonotic pathogen and is the leading cause of bacterial meningitis in adults in Vietnam. Systematic data on the antimicrobial susceptibility profiles of S. suis strains isolated from human cases are lacking. We studied antimicrobial resistance and associated resistance determinants in S. suis isolated from patients with meningitis in southern Vietnam. Methods S. suis strains isolated between 1997 and 2008 were investigated for their susceptibility to six antimicrobial agents. Strains were screened for the presence and expression of tetracycline and erythromycin resistance determinants and the association of tet(M) genes with Tn916- like transposons. The localization of tetracycline resistance gene tet(L) was determined by pulse field gel electrophoresis and Southern blotting. Results We observed a significant increase in resistance to tetracycline and chloramphenicol, which was concurrent with an increase in multi-drug resistance. In tetracycline resistance strains, we identified tet(M), tet(O), tet(W) and tet(L) and confirmed their expression. All tet(M) genes were associated with a Tn916-like transposon. The co-expression of tet(L) and other tetracycline resistance gene(s) encoding for ribosomal protection protein(s) was only detected in strains with a minimum inhibitory concentration (MIC) of tetracycline of ≥ 64 mg/L Conclusions We demonstrated that multi-drug resistance in S. suis causing disease in humans in southern Vietnam has increased over the 11-year period studied. We report the presence and expression of tet(L) in S. suis strains and our data suggest that co-expression of multiple genes encoding distinct mechanism is required for an MIC ≥ 64 mg/L to tetracycline. PMID:21208459

The antimicrobial resistance patterns and associated determinants in Streptococcus suis isolated from humans in southern Vietnam, 1997-2008.

PubMed

Hoa, Ngo T; Chieu, Tran T B; Nghia, Ho D T; Mai, Nguyen T H; Anh, Pham H; Wolbers, Marcel; Baker, Stephen; Campbell, James I; Chau, Nguyen V V; Hien, Tran T; Farrar, Jeremy; Schultsz, Constance

2011-01-06

Streptococcus suis is an emerging zoonotic pathogen and is the leading cause of bacterial meningitis in adults in Vietnam. Systematic data on the antimicrobial susceptibility profiles of S. suis strains isolated from human cases are lacking. We studied antimicrobial resistance and associated resistance determinants in S. suis isolated from patients with meningitis in southern Vietnam. S. suis strains isolated between 1997 and 2008 were investigated for their susceptibility to six antimicrobial agents. Strains were screened for the presence and expression of tetracycline and erythromycin resistance determinants and the association of tet(M) genes with Tn916- like transposons. The localization of tetracycline resistance gene tet(L) was determined by pulse field gel electrophoresis and Southern blotting. We observed a significant increase in resistance to tetracycline and chloramphenicol, which was concurrent with an increase in multi-drug resistance. In tetracycline resistance strains, we identified tet(M), tet(O), tet(W) and tet(L) and confirmed their expression. All tet(M) genes were associated with a Tn916-like transposon. The co-expression of tet(L) and other tetracycline resistance gene(s) encoding for ribosomal protection protein(s) was only detected in strains with a minimum inhibitory concentration (MIC) of tetracycline of ≥ 64 mg/L. We demonstrated that multi-drug resistance in S. suis causing disease in humans in southern Vietnam has increased over the 11-year period studied. We report the presence and expression of tet(L) in S. suis strains and our data suggest that co-expression of multiple genes encoding distinct mechanism is required for an MIC ≥ 64 mg/L to tetracycline.

Validation of quantitative and qualitative methods for detecting allergenic ingredients in processed foods in Japan.

PubMed

Sakai, Shinobu; Adachi, Reiko; Akiyama, Hiroshi; Teshima, Reiko

2013-06-19

A labeling system for food allergenic ingredients was established in Japan in April 2002. To monitor the labeling, the Japanese government announced official methods for detecting allergens in processed foods in November 2002. The official methods consist of quantitative screening tests using enzyme-linked immunosorbent assays (ELISAs) and qualitative confirmation tests using Western blotting or polymerase chain reactions (PCR). In addition, the Japanese government designated 10 μg protein/g food (the corresponding allergenic ingredient soluble protein weight/food weight), determined by ELISA, as the labeling threshold. To standardize the official methods, the criteria for the validation protocol were described in the official guidelines. This paper, which was presented at the Advances in Food Allergen Detection Symposium, ACS National Meeting and Expo, San Diego, CA, Spring 2012, describes the validation protocol outlined in the official Japanese guidelines, the results of interlaboratory studies for the quantitative detection method (ELISA for crustacean proteins) and the qualitative detection method (PCR for shrimp and crab DNAs), and the reliability of the detection methods.

Southern Pine Bark Beetle Guild

Treesearch

T. Evan Nebeker

2011-01-01

Dendroctonus frontalis (southern pine beetle), D. terebrans (black turpentine beetle), Ips avulsus (small southern pine engraver or four-spined engraver), I. grandicollis (five-spined engraver), and I. calligraphus (six-spined engraver) comprise the southern pine bark beetle guild. Often they are found sharing the same hosts in the Southeastern United States. They...

Southern cascades bioregion

Treesearch

Carl N. Skinner; Alan H. Taylor

2006-01-01

The Cascade Range extends from British Columbia, Canada, south to northern California where it meets the Sierra Nevada. The Southern Cascades bioregion in California is bounded on the west by the Sacramento Valley and the Klamath Mountains, and on the east by the Modoc Plateau and Great Basin. The bioregion encompasses the Southern Cascades section of Miles and Goudey...

Prediction of the optimum hybridization conditions of dot-blot-SNP analysis using estimated melting temperature of oligonucleotide probes.

PubMed

Shiokai, Sachiko; Kitashiba, Hiroyasu; Nishio, Takeshi

2010-08-01

Although the dot-blot-SNP technique is a simple cost-saving technique suitable for genotyping of many plant individuals, optimization of hybridization and washing conditions for each SNP marker requires much time and labor. For prediction of the optimum hybridization conditions for each probe, we compared T (m) values estimated from nucleotide sequences using the DINAMelt web server, measured T (m) values, and hybridization conditions yielding allele-specific signals. The estimated T (m) values were comparable to the measured T (m) values with small differences of less than 3 degrees C for most of the probes. There were differences of approximately 14 degrees C between the specific signal detection conditions and estimated T (m) values. Change of one level of SSC concentrations of 0.1, 0.2, 0.5, and 1.0x SSC corresponded to a difference of approximately 5 degrees C in optimum signal detection temperature. Increasing the sensitivity of signal detection by shortening the exposure time to X-ray film changed the optimum hybridization condition for specific signal detection. Addition of competitive oligonucleotides to the hybridization mixture increased the suitable hybridization conditions by 1.8. Based on these results, optimum hybridization conditions for newly produced dot-blot-SNP markers will become predictable.

Southern rim of Pacific Ocean basin: southern Andes to southern Alps

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

Dalziel, I.W.D.; Garrett, S.W.; Grunow, A.M.

1986-07-01

Between the southern Andes of Tierra del Fuego and the southern Alps of New Zealand lies the least accessible and geologically least explored part of the Pacific Ocean basin. A joint United Kingdom-United States project was initiated in 1983 to elucidate the geologic history and structure of the Pacific margin of Antarctica from the Antarctic Peninsula to Pine Island Bay at approximately lone. 105/sup 0/W. The first season (1983-1984) of this West Antarctic Tectonics Project was spent in the Ellsworth-Whitmore crustal block, and the second (1984-1985) in the Thurston Island crustal block. The project involves structural and general field geology,more » petrology, geochemistry, paleomagnetism, and airborne geophysics (magnetics and radar ice echo sounding). A final geologic season will be spent in the Pensacola Mountains of the Transantarctic Range in 1987-1988.« less

Emissions of Trace Gases and Particles from Savanna Fires in Southern Africa

NASA Technical Reports Server (NTRS)

Sinha, Parikhit; Hobbs, Peter V.; Yokelson, Robert J.; Bertschi, Isaac T.; Blake, Donald R.; Simpson, Isobel J.; Gao, Song; Kirchstetter, Thomas W.; Novakov, Tica

2003-01-01

Airborne measurements made on initial smoke from 10 savanna fires in southern Africa provide quantitative data on emissions of 50 gaseous and particulate species, including carbon dioxide, carbon monoxide, sulfur dioxide, nitrogen oxides, methane, ammonia, dimethyl sulfide, nonmethane organic compounds, halocarbons, gaseous organic acids, aerosol ionic components, carbonaceous aerosols, and condensation nuclei (CN). Measurements of several of the gaseous species by gas chromatography and Fourier transform infrared spectroscopy are compared. Emission ratios and emission factors are given for eight species that have not been reported previously for biomass burning of savanna in southern Africa (namely, dimethyl sulfide, methyl nitrate, five hydrocarbons, and particles with diameters from 0.1 to 3 microns). The emission factor that we measured for ammonia is lower by a factor of 4, and the emission factors for formaldehyde, hydrogen cyanide, and CN are greater by factors of about 3, 20, and 3 - 15, respectively, than previously reported values. The new emission factors are used to estimate annual emissions of these species from savanna fires in Africa and worldwide.

Profiling EGFR activity in head and neck squamous cell carcinoma by using a novel layered membrane Western blot technology.

PubMed

Patel, Vyomesh; Ramesh, Arun; Traicoff, June L; Baibakov, Galina; Emmert-Buck, Michael R; Gutkind, J Silvio; Knezevic, Vladimir

2005-05-01

Given the role of epidermal growth factor receptor (EGFR) in head and neck squamous cell carcinomas (HNSCC), several rational approaches have now been utilized to abrogate tyrosine kinase activity and its disengagement from downstream signal transducers. Monitoring the activity of these molecules could potentially be useful to determine not only drug efficacy but also to identify HNSCC patients most likely to benefit from this type of therapy. In this study we have used a novel high throughput multi-layered Western blotting (MLWestern) method that allows the detection of multiple proteins from a single experiment in order to characterize key components in the EGFR signaling pathway in HNSCC cells. Total and activated forms of EGFR and the downstream effectors, Erk and Akt were readily detected in HNSCC cells, where in the control cells (HaCaT) these proteins could only be detected in EGF stimulated cells. Results from conventional Western blot and MLWestern were comparable. Clustering analysis of protein expression revealed similarities in cellular response between some of the cell lines indicative of similarities in their biological response. The data indicate that MLWestern can be potentially applied to identify molecular targets that could be used for rational therapeutic intervention strategies.

Overview of the Southern San Andreas Fault Model

USGS Publications Warehouse

Weldon, Ray J.; Biasi, Glenn P.; Wills, Chris J.; Dawson, Timothy E.

2008-01-01

This appendix summarizes the data and methodology used to generate the source model for the southern San Andreas fault. It is organized into three sections, 1) a section by section review of the geological data in the format of past Working Groups, 2) an overview of the rupture model, and 3) a manuscript by Biasi and Weldon (in review Bulletin of the Seismological Society of America) that describes the correlation methodology that was used to help develop the ?geologic insight? model. The goal of the Biasi and Weldon methodology is to quantify the insight that went into developing all A faults; as such it is in concept consistent with all other A faults but applied in a more quantitative way. The most rapidly slipping fault and the only known source of M~8 earthquakes in southern California is the San Andreas fault. As such it plays a special role in the seismic hazard of California, and has received special attention in the current Working Group. The underlying philosophy of the current Working Group is to model the recurrence behavior of large, rapidly slipping faults like the San Andreas from observed data on the size, distribution and timing of past earthquakes with as few assumptions about underlying recurrence behavior as possible. In addition, we wish to carry the uncertainties in the data and the range of reasonable extrapolations from the data to the final model. To accomplish this for the Southern San Andreas fault we have developed an objective method to combine all of the observations of size, timing, and distribution of past earthquakes into a comprehensive set of earthquake scenarios that each represent a possible history of earthquakes for the past ~1400 years. The scenarios are then ranked according to their overall consistency with the data and then the frequencies of all of the ruptures permitted by the current Working Group?s segmentation model are calculated. We also present 30-yr conditional probabilities by segment and compare to previous

New Observationally-Based Metrics for the Analysis of Coupled Climate Model and Earth System Model Simulations of the Southern Ocean

NASA Astrophysics Data System (ADS)

Russell, J. L.

2014-12-01

The exchange of heat and carbon dioxide between the atmosphere and ocean are major controls on Earth's climate under conditions of anthropogenic forcing. The Southern Ocean south of 30°S, occupying just over ¼ of the surface ocean area, accounts for a disproportionate share of the vertical exchange of properties between the deep and surface waters of the ocean and between the surface ocean and the atmosphere; thus this region can be disproportionately influential on the climate system. Despite the crucial role of the Southern Ocean in the climate system, understanding of the particular mechanisms involved remains inadequate, and the model studies underlying many of these results are highly controversial. As part of the overall goal of working toward reducing uncertainties in climate projections, we present an analysis using new data/model metrics based on a unified framework of theory, quantitative datasets, and numerical modeling. These new metrics quantify the mechanisms, processes, and tendencies relevant to the role of the Southern Ocean in climate.

Novel approach to quantitative polymerase chain reaction using real-time detection: application to the detection of gene amplification in breast cancer.

PubMed

Bièche, I; Olivi, M; Champème, M H; Vidaud, D; Lidereau, R; Vidaud, M

1998-11-23

Gene amplification is a common event in the progression of human cancers, and amplified oncogenes have been shown to have diagnostic, prognostic and therapeutic relevance. A kinetic quantitative polymerase-chain-reaction (PCR) method, based on fluorescent TaqMan methodology and a new instrument (ABI Prism 7700 Sequence Detection System) capable of measuring fluorescence in real-time, was used to quantify gene amplification in tumor DNA. Reactions are characterized by the point during cycling when PCR amplification is still in the exponential phase, rather than the amount of PCR product accumulated after a fixed number of cycles. None of the reaction components is limited during the exponential phase, meaning that values are highly reproducible in reactions starting with the same copy number. This greatly improves the precision of DNA quantification. Moreover, real-time PCR does not require post-PCR sample handling, thereby preventing potential PCR-product carry-over contamination; it possesses a wide dynamic range of quantification and results in much faster and higher sample throughput. The real-time PCR method, was used to develop and validate a simple and rapid assay for the detection and quantification of the 3 most frequently amplified genes (myc, ccndl and erbB2) in breast tumors. Extra copies of myc, ccndl and erbB2 were observed in 10, 23 and 15%, respectively, of 108 breast-tumor DNA; the largest observed numbers of gene copies were 4.6, 18.6 and 15.1, respectively. These results correlated well with those of Southern blotting. The use of this new semi-automated technique will make molecular analysis of human cancers simpler and more reliable, and should find broad applications in clinical and research settings.

Description and quantitative analysis of the dentition of the southern thorny skate Amblyraja doellojuradoi.

PubMed

Delpiani, G; Spath, M C; Deli Antoni, M; Delpiani, M

2017-06-01

A description of the tooth morphology of 234 jaws from the southern thorny skate Amblyraja doellojuradoi in the south-west Atlantic Ocean is given. Seven rows of teeth were selected and length and width of each tooth in these rows were measured. It was found that functional series corresponds to the third teeth and the average width and length of these teeth were compared among jaws, maturity stages, sexes and rows. Generalized linear models were used to determine the subset of measures that most contribute to explain the variability between groups. It was observed that males have longer teeth than females, but the teeth of females are wider. These differences are attributed to reproductive behaviour, in which males bite females to hold them during copulation. This study provides a description of the teeth of A. doellojuradoi, supplying a valuable tool for identification of species. In addition, the establishment of the main variations observed in the dentition, improves the understanding of the species' biology. © 2017 The Fisheries Society of the British Isles.

Contribution of dot-blot assay to the diagnosis and management of myositis: a three-year practice at a university hospital centre.

PubMed

Martel, Clothilde; Vignaud, Guillaume; Liozon, Eric; Magy, Laurent; Gallouedec, Gael; Ly, Kim; Bezanahary, Holly; Cypierre, Anne; Lapébie, François-Xavier; Palat, Sylvain; Gondran, Guillaume; Jauberteau, Marie-Odile; Fauchais, Anne-Laure

2016-01-01

Idiopathic inflammatory myopathies (IIM) are heterogeneous autoimmune diseases with wide clinical spectrum that may lead to delayed diagnosis. The aim of this study was to examine the impact of IIM-specific dot-blot assay on diagnostic process of patients presenting with muscular or systemic symptoms evocating of IIM. We collected all the prescriptions of an IIM specific dot-blot assay (8 autoantigens including Jo-1, PL-7, PL-12, SRP, Mi-2, Ku, PM/Scl and Scl-70) over a 38-month period. 316 myositis dot-blot assays (MSD) were performed in 274 patients (156 women, mean age 53±10.6 years) referring for muscular and/or systemic symptoms suggesting IIM. The timing of dot prescription through the diagnostic process was highly variable: without (35%), concomitantly (16%) or after electromyographic studies (35%). Fifty-nine patients (22%) had IIM according to Bohan and Peter's criteria. Among them, 29 (49%) had positive dot (8 Jo-1, 6 PM-Scl, 5 PL-12, 5 SRP, 2 Mi-2, 2 PL-7 and 1 Ku). Various other diagnoses were performed including 35 autoimmune disease or granulomatosis (12%), 19 inflammatory rheumatic disease (7%), 16 non inflammatory muscular disorders (6%), 10 drug-induced myalgia (4%), 11 infectious myositis (4%). Except 11 borderline SRP results and one transient PM-Scl, MSD was positive only in one case of IIM. Dot allowed clinicians to correct diagnosis in 4 cases and improved the diagnosis of IIM subtypes in 4 cases. This study reflects the interest of myositis dot in the rapid diagnosis process of patients with non-specific muscular symptoms leading to various diagnoses including IIM.

Characterization of Nora Virus Structural Proteins via Western Blot Analysis.

PubMed

Ericson, Brad L; Carlson, Darby J; Carlson, Kimberly A

2016-01-01

Nora virus is a single stranded RNA picorna-like virus with four open reading frames (ORFs). The coding potentials of the ORFs are not fully characterized, but ORF3 and ORF4 are believed to encode the capsid proteins (VP3, VP4a, VP4b, and VP4c) comprising the virion. To determine the polypeptide composition of Nora virus virions, polypeptides from purified virus were compared to polypeptides detected in Nora virus infected Drosophila melanogaster. Nora virus was purified from infected flies and used to challenge mice for the production of antisera. ORF3, ORF4a, ORF4b, and ORF4c were individually cloned and expressed in E. coli; resultant recombinant proteins purified and were used to make monospecific antisera. Antisera were evaluated via Western blot against whole virus particles and Nora virus infected fly lysates. Viral purification yielded two particle types with densities of ~1.31 g/mL (empty particles) and ~1.33 g/mL (complete virions). Comparison of purified virus polypeptide composition to Nora virus infected D. melanogaster lysate showed the number of proteins in infected cell lysates is less than purified virus. Our results suggest the virion is composed of 6 polypeptides, VP3, VP4a, two forms of VP4b, and two forms of VP4c. This polypeptide composition is similar to other small RNA insect viruses.

Characterization of Nora Virus Structural Proteins via Western Blot Analysis

PubMed Central

Ericson, Brad L.; Carlson, Darby J.

2016-01-01

Nora virus is a single stranded RNA picorna-like virus with four open reading frames (ORFs). The coding potentials of the ORFs are not fully characterized, but ORF3 and ORF4 are believed to encode the capsid proteins (VP3, VP4a, VP4b, and VP4c) comprising the virion. To determine the polypeptide composition of Nora virus virions, polypeptides from purified virus were compared to polypeptides detected in Nora virus infected Drosophila melanogaster. Nora virus was purified from infected flies and used to challenge mice for the production of antisera. ORF3, ORF4a, ORF4b, and ORF4c were individually cloned and expressed in E. coli; resultant recombinant proteins purified and were used to make monospecific antisera. Antisera were evaluated via Western blot against whole virus particles and Nora virus infected fly lysates. Viral purification yielded two particle types with densities of ~1.31 g/mL (empty particles) and ~1.33 g/mL (complete virions). Comparison of purified virus polypeptide composition to Nora virus infected D. melanogaster lysate showed the number of proteins in infected cell lysates is less than purified virus. Our results suggest the virion is composed of 6 polypeptides, VP3, VP4a, two forms of VP4b, and two forms of VP4c. This polypeptide composition is similar to other small RNA insect viruses. PMID:27298753

Muscadine Grape Skin Extract Induces an Unfolded Protein Response-Mediated Autophagy in Prostate Cancer Cells: A TMT-Based Quantitative Proteomic Analysis

PubMed Central

Burton, Liza J.; Rivera, Mariela; Hawsawi, Ohuod; Zou, Jin; Hudson, Tamaro; Wang, Guangdi; Zhang, Qiang; Cubano, Luis; Boukli, Nawal; Odero-Marah, Valerie

2016-01-01

Muscadine grape skin extract (MSKE) is derived from muscadine grape (Vitis rotundifolia), a common red grape used to produce red wine. Endoplasmic reticulum (ER) stress activates the unfolded protein response (UPR) that serves as a survival mechanism to relieve ER stress and restore ER homeostasis. However, when persistent, ER stress can alter the cytoprotective functions of the UPR to promote autophagy and cell death. Although MSKE has been documented to induce apoptosis, it has not been linked to ER stress/UPR/autophagy. We hypothesized that MSKE may induce a severe ER stress response-mediated autophagy leading to apoptosis. As a model, we treated C4-2 prostate cancer cells with MSKE and performed a quantitative Tandem Mass Tag Isobaric Labeling proteomic analysis. ER stress response, autophagy and apoptosis were analyzed by western blot, acridine orange and TUNEL/Annexin V staining, respectively. Quantitative proteomics analysis indicated that ER stress response proteins, such as GRP78 were greatly elevated following treatment with MSKE. The up-regulation of pro-apoptotic markers PARP, caspase-12, cleaved caspase-3, -7, BAX and down-regulation of anti-apoptotic marker BCL2 was confirmed by Western blot analysis and apoptosis was visualized by increased TUNEL/Annexin V staining upon MSKE treatment. Moreover, increased acridine orange, and LC3B staining was detected in MSKE-treated cells, suggesting an ER stress/autophagy response. Finally, MSKE-mediated autophagy and apoptosis was antagonized by co-treatment with chloroquine, an autophagy inhibitor. Our results indicate that MSKE can elicit an UPR that can eventually lead to apoptosis in prostate cancer cells. PMID:27755556

Muscadine Grape Skin Extract Induces an Unfolded Protein Response-Mediated Autophagy in Prostate Cancer Cells: A TMT-Based Quantitative Proteomic Analysis.

PubMed

Burton, Liza J; Rivera, Mariela; Hawsawi, Ohuod; Zou, Jin; Hudson, Tamaro; Wang, Guangdi; Zhang, Qiang; Cubano, Luis; Boukli, Nawal; Odero-Marah, Valerie

2016-01-01

Muscadine grape skin extract (MSKE) is derived from muscadine grape (Vitis rotundifolia), a common red grape used to produce red wine. Endoplasmic reticulum (ER) stress activates the unfolded protein response (UPR) that serves as a survival mechanism to relieve ER stress and restore ER homeostasis. However, when persistent, ER stress can alter the cytoprotective functions of the UPR to promote autophagy and cell death. Although MSKE has been documented to induce apoptosis, it has not been linked to ER stress/UPR/autophagy. We hypothesized that MSKE may induce a severe ER stress response-mediated autophagy leading to apoptosis. As a model, we treated C4-2 prostate cancer cells with MSKE and performed a quantitative Tandem Mass Tag Isobaric Labeling proteomic analysis. ER stress response, autophagy and apoptosis were analyzed by western blot, acridine orange and TUNEL/Annexin V staining, respectively. Quantitative proteomics analysis indicated that ER stress response proteins, such as GRP78 were greatly elevated following treatment with MSKE. The up-regulation of pro-apoptotic markers PARP, caspase-12, cleaved caspase-3, -7, BAX and down-regulation of anti-apoptotic marker BCL2 was confirmed by Western blot analysis and apoptosis was visualized by increased TUNEL/Annexin V staining upon MSKE treatment. Moreover, increased acridine orange, and LC3B staining was detected in MSKE-treated cells, suggesting an ER stress/autophagy response. Finally, MSKE-mediated autophagy and apoptosis was antagonized by co-treatment with chloroquine, an autophagy inhibitor. Our results indicate that MSKE can elicit an UPR that can eventually lead to apoptosis in prostate cancer cells.

Isolation and characterization of cDNA clones for human erythrocyte beta-spectrin.

PubMed Central

Prchal, J T; Morley, B J; Yoon, S H; Coetzer, T L; Palek, J; Conboy, J G; Kan, Y W

1987-01-01

Spectrin is an important structural component of the membrane skeleton that underlies and supports the erythrocyte plasma membrane. It is composed of nonidentical alpha (Mr 240,000) and beta (Mr 220,000) subunits, each of which contains multiple homologous 106-amino acid segments. We report here the isolation and characterization of a human erythroid-specific beta-spectrin cDNA clone that encodes parts of the beta-9 through beta-12 repeat segments. This cDNA was used as a hybridization probe to assign the beta-spectrin gene to human chromosome 14 and to begin molecular analysis of the gene and its mRNA transcripts. RNA transfer blot analysis showed that the reticulocyte beta-spectrin mRNA is 7.8 kilobases in length. Southern blot analysis of genomic DNA revealed the presence of restriction fragment length polymorphisms (RFLPs) within the beta-spectrin gene locus. The isolation of human spectrin cDNA probes and the identification of closely linked RFLPs will facilitate analysis of mutant spectrin genes causing congenital hemolytic anemias associated with quantitative and qualitative spectrin abnormalities. Images PMID:3478706

Rosette Assay: Highly Customizable Dot-Blot for SH2 Domain Screening.

PubMed

Ng, Khong Y; Machida, Kazuya

2017-01-01

With a growing number of high-throughput studies, structural analyses, and availability of protein-protein interaction databases, it is now possible to apply web-based prediction tools to SH2 domain-interactions. However, in silico prediction is not always reliable and requires experimental validation. Rosette assay is a dot blot-based reverse-phase assay developed for the assessment of binding between SH2 domains and their ligands. It is conveniently customizable, allowing for low- to high-throughput analysis of interactions between various numbers of SH2 domains and their ligands, e.g., short peptides, purified proteins, and cell lysates. The binding assay is performed in a 96-well plate (MBA or MWA apparatus) in which a sample spotted membrane is incubated with up to 96 labeled SH2 domains. Bound domains are detected and quantified using a chemiluminescence or near-infrared fluorescence (IR) imaging system. In this chapter, we describe a practical protocol for rosette assay to assess interactions between synthesized tyrosine phosphorylated peptides and a library of GST-tagged SH2 domains. Since the methodology is not confined to assessment of SH2-pTyr interactions, rosette assay can be broadly utilized for ligand and drug screening using different protein interaction domains or antibodies.

Comparison of Multispot EIA with Western blot for confirmatory serodiagnosis of HIV.

PubMed

Torian, Lucia V; Forgione, Lisa A; Punsalang, Amado E; Pirillo, Robert E; Oleszko, William R

2011-12-01

Recent improvements in the sensitivity of immunoassays (IA) used for HIV screening, coupled with increasing recognition of the importance of rapid point-of-care testing, have led to proposals to adjust the algorithm for serodiagnosis of HIV so that screening and confirmation can be performed using a dual or triple IA sequence that does not require Western blotting for confirmation. One IA that has been proposed as a second or confirmatory test is the Bio-Rad Multispot(®) Rapid HIV-1/HIV-2 Test. This test would have the added advantage of differentiating between HIV-1 and HIV-2 antibodies. To compare the sensitivity and type-specificity of an algorithm combining a 3rd generation enzyme immunoassay (EIA) followed by a confirmatory Multispot with the conventional algorithm that combines a 3rd generation EIA (Bio-Rad GS HIV-1/HIV-2 Plus O EIA) followed by confirmatory Western blot (Bio-Rad GS HIV-1 WB). 8760 serum specimens submitted for HIV testing to the New York City Public Health Laboratory between May 22, 2007, and April 30, 2010, tested repeatedly positive on 3rd generation HIV-1-2+O EIA screening and received parallel confirmatory testing by WB and Multispot (MS). 8678/8760 (99.1%) specimens tested WB-positive; 82 (0.9%) tested WB-negative or indeterminate (IND). 8690/8760 specimens (99.2%) tested MS-positive, of which 14 (17.1%) had been classified as negative or IND by WB. Among the HIV-1 WB-positive specimens, MS classified 26 (0.29%) as HIV-2. Among the HIV-1 WB negative and IND, MS detected 12 HIV-2. MS detected an additional 14 HIV-1 infections among WB negative or IND specimens, differentiated 26 HIV-1 WB positives as HIV-2, and detected 12 additional HIV-2 infections among WB negative/IND. A dual 3rd generation EIA algorithm incorporating MS had equivalent HIV-1 sensitivity to the 3rd generation EIA-WB algorithm and had the added advantage of detecting 12 HIV-2 specimens that were not HIV-1 WB cross-reactors. In this series an algorithm using EIA

Capillary nano-immunoassays: advancing quantitative proteomics analysis, biomarker assessment, and molecular diagnostics.

PubMed

Chen, Jin-Qiu; Wakefield, Lalage M; Goldstein, David J

2015-06-06

There is an emerging demand for the use of molecular profiling to facilitate biomarker identification and development, and to stratify patients for more efficient treatment decisions with reduced adverse effects. In the past decade, great strides have been made to advance genomic, transcriptomic and proteomic approaches to address these demands. While there has been much progress with these large scale approaches, profiling at the protein level still faces challenges due to limitations in clinical sample size, poor reproducibility, unreliable quantitation, and lack of assay robustness. A novel automated capillary nano-immunoassay (CNIA) technology has been developed. This technology offers precise and accurate measurement of proteins and their post-translational modifications using either charge-based or size-based separation formats. The system not only uses ultralow nanogram levels of protein but also allows multi-analyte analysis using a parallel single-analyte format for increased sensitivity and specificity. The high sensitivity and excellent reproducibility of this technology make it particularly powerful for analysis of clinical samples. Furthermore, the system can distinguish and detect specific protein post-translational modifications that conventional Western blot and other immunoassays cannot easily capture. This review will summarize and evaluate the latest progress to optimize the CNIA system for comprehensive, quantitative protein and signaling event characterization. It will also discuss how the technology has been successfully applied in both discovery research and clinical studies, for signaling pathway dissection, proteomic biomarker assessment, targeted treatment evaluation and quantitative proteomic analysis. Lastly, a comparison of this novel system with other conventional immuno-assay platforms is performed.

The role of diatom resting spores in pelagic-benthic coupling in the Southern Ocean

NASA Astrophysics Data System (ADS)

Rembauville, Mathieu; Blain, Stéphane; Manno, Clara; Tarling, Geraint; Thompson, Anu; Wolff, George; Salter, Ian

2018-05-01

Natural iron fertilization downstream of Southern Ocean island plateaus supports large phytoplankton blooms and promotes carbon export from the mixed layer. In addition to sequestering atmospheric CO2, the biological carbon pump also supplies organic matter (OM) to deep-ocean ecosystems. Although the total flux of OM arriving at the seafloor sets the energy input to the system, the chemical nature of OM is also of significance. However, a quantitative framework linking ecological flux vectors to OM composition is currently lacking. In the present study we report the lipid composition of export fluxes collected by five moored sediment traps deployed in contrasting productivity regimes of Southern Ocean island systems (Kerguelen, Crozet and South Georgia) and compile them with quantitative data on diatom and faecal pellet fluxes. At the three naturally iron-fertilized sites, the relative contribution of labile lipids (mono- and polyunsaturated fatty acids, unsaturated fatty alcohols) is 2-4 times higher than at low productivity sites. There is a strong attenuation of labile components as a function of depth, irrespective of productivity. The three island systems also display regional characteristics in lipid export. An enrichment of zooplankton dietary sterols, such as C27Δ5, at South Georgia is consistent with high zooplankton and krill biomass in the region and the importance of faecal pellets to particulate organic carbon (POC) flux. There is a strong association of diatom resting spore fluxes that dominate productive flux regimes with energy-rich unsaturated fatty acids. At the Kerguelen Plateau we provide a statistical framework to link seasonal variation in ecological flux vectors and lipid composition over a complete annual cycle. Our analyses demonstrate that ecological processes in the upper ocean, e.g. resting spore formation and grazing, not only impact the magnitude and stoichiometry of the Southern Ocean biological pump, but also regulate the

Southern pulpwood production, 1982

Treesearch

James F. Rosson

1982-01-01

The annual report of southern pulpwood production is based on a canvass of all pulpmills that fall within the 12 states of the Southern and Southeastern Forest Experiment Station regions. Neighboring experiment stations exchange information on mills located beyond the region but utilizing wood from the region.

Quantitative and temporal proteome analysis of butyrate-treated colorectal cancer cells.

PubMed

Tan, Hwee Tong; Tan, Sandra; Lin, Qingsong; Lim, Teck Kwang; Hew, Choy Leong; Chung, Maxey C M

2008-06-01

Colorectal cancer is one of the most common cancers in developed countries, and its incidence is negatively associated with high dietary fiber intake. Butyrate, a short-chain fatty acid fermentation by-product of fiber induces cell maturation with the promotion of growth arrest, differentiation, and/or apoptosis of cancer cells. The stimulation of cell maturation by butyrate in colonic cancer cells follows a temporal progression from the early phase of growth arrest to the activation of apoptotic cascades. Previously we performed two-dimensional DIGE to identify differentially expressed proteins induced by 24-h butyrate treatment of HCT-116 colorectal cancer cells. Herein we used quantitative proteomics approaches using iTRAQ (isobaric tags for relative and absolute quantitation), a stable isotope labeling methodology that enables multiplexing of four samples, for a temporal study of HCT-116 cells treated with butyrate. In addition, cleavable ICAT, which selectively tags cysteine-containing proteins, was also used, and the results complemented those obtained from the iTRAQ strategy. Selected protein targets were validated by real time PCR and Western blotting. A model is proposed to illustrate our findings from this temporal analysis of the butyrate-responsive proteome that uncovered several integrated cellular processes and pathways involved in growth arrest, apoptosis, and metastasis. These signature clusters of butyrate-regulated pathways are potential targets for novel chemopreventive and therapeutic drugs for treatment of colorectal cancer.

The Recent Rise of Southern Magazines.

ERIC Educational Resources Information Center

Hynds, Ernest C.

During the past decade states in the southern United States have produced an increasing percentage of the nation's "city" magazines. Three magazines illustrate what the southern metropolitan magazines are doing to serve their readers and their communities. "Southern Living" provides information about its readers' interests as…

Southern pulpwood production, 1993

Treesearch

Michael Howell; Andrew J. Hartsell

1993-01-01

In 1993, Southern pulpwood production declined 2 percent to 66.3 million cords. Roundwood production decreased 2 percent to 46.3 million cords, and wood residue production fell 4 percent to 19.9 million cords. Pulping capacity of the 102 Southern pulpmills was 132,327 tons per day.

ATLAS: An advanced PCR-method for routine visualization of telomere length in Saccharomyces cerevisiae.

PubMed

Zubko, Elena I; Shackleton, Jennifer L; Zubko, Mikhajlo K

2016-12-01

Measuring telomere length is essential in telomere biology. Southern blot hybridization is the predominant method for measuring telomere length in the genetic model Saccharomyces cerevisiae. We have further developed and refined a telomere PCR approach, which was rarely used previously (mainly in specific telomeric projects), into a robust method allowing direct visualisation of telomere length differences in routine experiments with S. cerevisiae, and showing a strong correlation of results with data obtained by Southern blot hybridization. In this expanded method denoted as ATLAS (A-dvanced T-elomere L-ength A-nalysis in S. cerevisiae), we have introduced: 1) set of new primers annealing with high specificity to telomeric regions on five different chromosomes; 2) new approach for designing reverse telomere primers that is based on the ligation of an adaptor of a fixed size to telomeric ends. ATLAS can be used at the scale of individual assays and high-throughput approaches. This simple, time/cost-effective and reproducible methodology will complement Southern blot hybridization and facilitate further progress in telomere research. Copyright © 2016 Elsevier B.V. All rights reserved.

Interaction of murine intestinal mast cell proteinase with inhibitors (serpins) in blood; analysis by SDS-PAGE and western blotting.

PubMed Central

Irvine, J; Newlands, G F; Huntley, J F; Miller, H R

1990-01-01

The interaction of mouse intestinal mast cell proteinase (IMCP) with serine proteinase inhibitors (serpins) in blood was analysed: (i) by examining the capacity of the inhibitors in blood to block the binding of the irreversible serine esterase inhibitor [3H]diisopropyl fluorophosphate (DFP); (ii) by Western blotting. The binding of [3H]DFP to IMCP was blocked very rapidly by inhibitors in mouse serum and, by Western blotting, this inhibition was associated with the appearance of a 73,000 MW proteinase/inhibitor complex together with a series of higher (greater than 100,000) MW complexes. IMCP was not dissociated from these complexes when electrophoresed under reducing conditions, although prior heat treatment of mouse serum (60 for 30-160 min) abolished the formation of all proteinase/inhibitor complexes. Similarly, the activity of a 48,000 MW inhibitor of chymotrypsin was abolished by heat treatment. A titration experiment established that between 0.5 and 5 mg IMCP were inhibited per ml of serum. The properties and MW of the IMCP inhibitor complexes are typical of serpins and suggest that IMCP secreted during intestinal immunological reactions would be rapidly and irreversibly inactivated by plasma-derived inhibitors. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:2312150

Southern pulpwood production, 1990

Treesearch

John S. Vissage; Patrick E. Miller

1992-01-01

In 1990, Southern pulpwood production exceeded 65 million cords, roundwood production increased 9 percent to 45.6 million cords, and wood residue production increased 2 percent to 19.4 million cords. The pulping capacity of the 103 Southern pulpmills was 129,290 tons per day. One pulpmill was under construction.

Comparison of a serum indirect fluorescent antibody test with two Western blot tests for the diagnosis of equine protozoal myeloencephalitis.

PubMed

Duarte, Paulo C; Daft, Barbara M; Conrad, Patricia A; Packham, Andrea E; Gardner, Ian A

2003-01-01

A serum indirect fluorescent antibody test (IFAT) was compared with a Western blot (WB) and a modified Western blot (mWB) for diagnosis of equine protozoal myeloencephalitis (EPM). Using receiver-operating characteristic (ROC) analysis, the area under the curve of the IFAT was greater than the areaunder the curves of the WB and the mWB (P = 0.025 and P = 0.044, respectively). There was no statistically significant difference between the areas under the curves of the WBs (P > 0.05). On the basis of an arbitrarily chosen cut-off titer for a positive test result of 1:80 for the IFAT and interpreting weak positive WB results as positive test results, the sensitivities and 95% confidence intervals (CI) of all 3 tests were identical and equal to 88.9% (51.8-99.7%). The specificities and 95% CIs of the IFAT, WB, and mWB test were 100% (91-100%), 87.2% (72.6-95.7%), and 69.2% (52.4-83%), respectively. The overall accuracy of the IFAT was shown to be better than that of the WBs and, therefore, the test has potential for use in the diagnosis of EPM caused by Sarcocystis neurona.

Southern Pine Beetle II

Treesearch

R. N. Coulson; Kier Klepzig

2011-01-01

The knowledge base for the southern pine beetle, Dendroctonus frontalis Zimmermann (Coleoptera: Curculionidae) has increased dramatically since the last comprehensive and interpretative summary (Thatcher and others 1980). This insect continues to be a significant pest affecting the forest environment of the Southern US and adjoining states and it is also the subject of...

Southern pulpwood production, 1974

Treesearch

Daniel F. Bertelson

1975-01-01

Southern pulpwood production in 1974 reached a record high of 49,102,144 cords, a 4-percent increase over the previous year. Daily pulping capacity at the 114 southern pulpmills rose 3 per- cent to 94,982 tons; the increase resulted from expansion of existing facilities and the opening of three new mills.

Rapid and quantitative detection of zoonotic influenza A virus infection utilizing coumarin-derived dendrimer-based fluorescent immunochromatographic strip test (FICT).

PubMed

Yeo, Seon-Ju; Huong, Dinh Thi; Hong, Nguyen Ngoc; Li, Chun-Ying; Choi, Kyunghan; Yu, Kyoungsik; Choi, Du-Young; Chong, Chom-Kyu; Choi, Hak Soo; Mallik, Shyam Kumar; Kim, Hak Sung; Sung, Haan Woo; Park, Hyun

2014-01-01

Great efforts have been made to develop robust signal-generating fluorescence materials which will help in improving the rapid diagnostic test (RDT) in terms of sensitivity and quantification. In this study, we developed coumarin-derived dendrimer-based fluorescent immunochromatographic strip test (FICT) assay with enhanced sensitivity as a quantitative diagnostic tool in typical RDT environments. The accuracy of the proposed FICT was compared with that of dot blot immunoassay techniques and conventional RDTs. Through conjugation of coumarin-derived dendrimers with latex beads, fluorescent emission covering broad output spectral ranges was obtained which provided a distinct advantage of easy discrimination of the fluorescent emission of the latex beads with a simple insertion of a long-pass optical filter away from the excitation wavelength. The newly developed FICT assay was able to detect 100 ng/10 μL of influenza A nucleoprotein (NP) antigen within 5 minutes, which corresponded to 2.5-fold higher sensitivity than that of the dot blot immunoassay or conventional RDTs. Moreover, the FICT assay was confirmed to detect at least four avian influenza A subtypes (H5N3, H7N1, H7N7, and H9N2). On applying the FICT to the clinical swab samples infected with respiratory viruses, our FICT assay was confirmed to differentiate influenza H1N1 infection from other respiratory viral diseases. These data demonstrate that the proposed FICT assay is able to detect zoonotic influenza A viruses with a high sensitivity, and it enables the quantitation of the infection intensity by providing the numerical diagnostic values; thus demonstrating enhanced detectability of influenza A viruses.

Distribution of a macaque immunosuppressive type D retrovirus in neural, lymphoid, and salivary tissues

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

Lackner, A.A.; Rodriguez, M.H.; Bush, C.E.

1988-06-01

Simian acquired immune deficiency syndrome (SAIDS) in rhesus macaques (Macaca mulatta) at the California Primate Research Center is caused by a type D retrovirus designated SAIDS retrovirus serotype 1 (SRV-1). This syndrome is characterized by profound immunosuppression and death associated with opportunistic infections. Neurologic signs and lesions have not been described as part of this syndrome. The distribution of SRV-1 in the salivary glands, lymph nodes, spleens, thymuses, and brains of eight virus-infected rhesus macaques was examined by immunohistochemistry. Electron microscopy, in situ RNA hybridization, and Southern blot hybridization were also performed on selected tissues to detect viral particles, RNA,more » and DNA, respectively. In seven of eight SRV-1-infected animals, the transmembrane envelope glycoprotein (gp20) of SRV-1 was present in three or more tissues, but never in the brain. In the remaining animal, no viral antigen was detected in any tissue. In this same group of animals, viral nucleic acid was detected in the lymph nodes of six of six animals by Southern blot hybridization, in the salivary glands of two of five animals by both Southern blot and in situ hybridizations, and, surprisingly, in the brains of three of three animals by Southern blot and of three of five animals by in situ hybridization, including the one animal in which viral gp20 was undetectable. None of these animals had neurologic signs or lesions. The detection of viral nucleic acid in the absence of viral antigen in the brain suggests latent SRV-1 infection of the central nervous system.« less

The Southern Annual Forest Inventory System

Treesearch

Gregory A. Reams; Paul C. van Deusen

1999-01-01

The Southern Annual Forest Inventory System (SAFIS) is in various stages of implementation in 7 of the 13 southern states serviced by the Southern Research Station. The SAFIS design is an interpenetrating design where the n units (1/6 acre plots) are divided into k = 5 panels, each panel containing m = n...

Southern pulpwood production, 1989

Treesearch

Cecil C. Hutchins

1991-01-01

This annual publication issued in alternate years by the Southeastern and Southern Forest Experiment stations, is based on 100-percent canvass of all pulpmills in the 12 Southern States from Virginia to Texas. The movement of raw material between regions is obtained by exchanging information with other regional Experiment Stations. Roundwood volumes are recorded in...

Meteor showers of the southern hemisphere

NASA Astrophysics Data System (ADS)

Molau, Sirko; Kerr, Steve

2014-04-01

We present the results of an exhaustive meteor shower search in the southern hemisphere. The underlying data set is a subset of the IMO Video Meteor Database comprising 50,000 single station meteors obtained by three Australian cameras between 2001 and 2012. The detection technique was similar to previous single station analysis. In the data set we find 4 major and 6 minor northern hemisphere meteor showers, and 12 segments of the Antihelion source (including the Northern and Southern Taurids and six streams from the MDC working list). We present details for 14 southern hemisphere showers plus the Centaurid and Puppid-Velid complex, with the η Aquariids and the Southern δ Aquariids being the strongest southern showers. Two of the showers (θ^2 Sagittariids and τ Cetids) were previously unknown and have received preliminary designations by the MDC. Overall we find that the fraction of southern meteor showers south of -30deg declination (roughly 25%) is clearly smaller than the fraction of northern meteor showers north of +30deg declination (more than 50%) obtained in our previous analysis.

Southern Ocean carbon-wind stress feedback

NASA Astrophysics Data System (ADS)

Bronselaer, Ben; Zanna, Laure; Munday, David R.; Lowe, Jason

2018-02-01

The Southern Ocean is the largest sink of anthropogenic carbon in the present-day climate. Here, Southern Ocean pCO2 and its dependence on wind forcing are investigated using an equilibrium mixed layer carbon budget. This budget is used to derive an expression for Southern Ocean pCO2 sensitivity to wind stress. Southern Ocean pCO2 is found to vary as the square root of area-mean wind stress, arising from the dominance of vertical mixing over other processes such as lateral Ekman transport. The expression for pCO2 is validated using idealised coarse-resolution ocean numerical experiments. Additionally, we show that increased (decreased) stratification through surface warming reduces (increases) the sensitivity of the Southern Ocean pCO2 to wind stress. The scaling is then used to estimate the wind-stress induced changes of atmospheric pCO_2 in CMIP5 models using only a handful of parameters. The scaling is further used to model the anthropogenic carbon sink, showing a long-term reversal of the Southern Ocean sink for large wind stress strength.

Identification of a chicken (Gallus gallus) endogenous reference gene (Actb) and its application in meat adulteration.

PubMed

Xiang, Wenjin; Shang, Ying; Wang, Qin; Xu, Yuancong; Zhu, Pengyu; Huang, Kunlun; Xu, Wentao

2017-11-01

The genes commonly used to determine meat species are mainly mitochondrial, but the copy numbers of such genes are high, meaning they cannot be accurately quantified. In this paper, for the first time, the chromosomal gene Actb was selected as an endogenous reference gene for chicken species. It was assayed in four different chicken varieties and 16 other species using both qualitative and quantitative PCR. No amplification of the Actb gene was found in species other than chicken and no allelic variations were detected in chicken. Southern blot and digital-PCR confirmed the Actb gene was present as a single copy in the chicken genome. The quantitative detection limit was 10pg of DNA, which is equivalent to eight copies. All experiments indicated that the Actb gene is a useful endogenous reference gene for chicken, and provides a convenient and accurate approach for detection of chicken in feed and food. Copyright © 2017 Elsevier Ltd. All rights reserved.

Southern pulpwood production, 1991

Treesearch

Michael Howell

1993-01-01

In 1991, Southern pulpwood production climbed 1 percent, to 65.08 million cords. Roundwood production increased 2 percent to 46.6 million cords, and wood residue production fell 5 percent to 18.4 million cords. One new pulpmill began operating in the region. Pulping capacity of the 104 southern pulpmill was 133.331 tons per day.

Southern pulpwood production, 1984

Treesearch

Dennis M. May

1986-01-01

This publication presents the findings of 100-percent canvass of all wood-using pulpmills drawing roundwood or wood residues from the 12 southern states (fig. 1). Canvass data are compiled annually and analyzed on an alternating basis by personnel of the Forest Inventory and Analysis Units of the Southern and southeastern Forest Experiment Stations. All production...

The archaeological evidence for the appearance of pastoralism and farming in southern Africa

PubMed Central

2018-01-01

This paper is a response to the growing reference to archaeological evidence by linguists and geneticists interested in the spread of early farmers and pastoralists in southern Africa. It presents two databases. The first contains the archaeological evidence for pastoralism and farming in southern Africa, for the period 550 BC to AD 1050. This is the first time that the seven different types of archaeological evidence that have traditionally been used to identify both spread events are presented together at this scale. This was stimulated by our interest in investigating the antiquity of an early ‘Iron Age package’ relative to the spread of single archaeological traits. The analysis shows that the package appears approximately 700 years after sites containing pottery, cattle and sheep, without agriculture, appear in the drier parts of the sub-continent. It post-dates the appearance of earlier sites with pottery associated with farmers, metal-working and cultivation in the eastern half of the sub-continent. While poor preservation undoubtedly explains the absence of some parts of the package, the analysis suggests that other explanations should be considered. The second database is a quantitative, spatial study of archaeological publications on southern African farming and pastoralism for the period 1950 to 2016, covering the same geographical area and archaeological timeframe. This is presented as a proxy for research-intensive areas in attempt to show the gaps in archaeological fieldwork and knowledge. PMID:29902271

Geologic Controls of Hydrocarbon Occurrence in the Southern Appalachian Basin in Eastern Tennessee, Southwestern Virginia, Eastern Kentucky, and Southern West Virginia

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

Robert D. Hatcher

2003-05-31

This report summarizes the first-year accomplishments of a three-year program to investigate the geologic controls of hydrocarbon occurrence in the southern Appalachian basin in eastern Tennessee, southwestern Virginia, eastern Kentucky, and southern West Virginia. The project: (1) employs the petroleum system approach to understand the geologic controls of hydrocarbons; (2) attempts to characterize the T-P parameters driving petroleum evolution; (3) attempts to obtain more quantitative definitions of reservoir architecture and identify new traps; (4) is working with USGS and industry partners to develop new play concepts and geophysical log standards for subsurface correlation; and (5) is geochemically characterizing the hydrocarbonsmore » (cooperatively with USGS). First-year results include: (1) meeting specific milestones (determination of thrust movement vectors, fracture analysis, and communicating results at professional meetings and through publication). All milestones were met. Movement vectors for Valley and Ridge thrusts were confirmed to be west-directed and derived from pushing by the Blue Ridge thrust sheet, and fan about the Tennessee salient. Fracture systems developed during Paleozoic, Mesozoic, and Cenozoic to Holocene compressional and extensional tectonic events, and are more intense near faults. Presentations of first-year results were made at the Tennessee Oil and Gas Association meeting (invited) in June, 2003, at a workshop in August 2003 on geophysical logs in Ordovician rocks, and at the Eastern Section AAPG meeting in September 2003. Papers on thrust tectonics and a major prospect discovered during the first year are in press in an AAPG Memoir and published in the July 28, 2003, issue of the Oil and Gas Journal. (2) collaboration with industry and USGS partners. Several Middle Ordovician black shale samples were sent to USGS for organic carbon analysis. Mississippian and Middle Ordovician rock samples were collected by John Repetski

Holocene local forest history at two sites in Småland, southern Sweden - insights from quantitative reconstructions using the Landscape Reconstruction Algorithm

NASA Astrophysics Data System (ADS)

Cui, Qiaoyu; Gaillard, Marie-José; Lemdahl, Geoffrey; Olsson, Fredrik; Sugita, Shinya

2010-05-01

Quantitative reconstruction of past vegetation using fossil pollen was long very problematic. It is well known that pollen percentages and pollen accumulation rates do not represent vegetation abundance properly because pollen values are influenced by many factors of which inter-taxonomic differences in pollen productivity and vegetation structure are the most important ones. It is also recognized that pollen assemblages from large sites (lakes or bogs) record the characteristics of the regional vegetation, while pollen assemblages from small sites record local features. Based on the theoretical understanding of the factors and mechanisms that affect pollen representation of vegetation, Sugita (2007a and b) proposed the Landscape Reconstruction Algorithm (LRA) to estimate vegetation abundance in percentage cover for well defined spatial scales. The LRA includes two models, REVEALS and LOVE. REVEALS estimates regional vegetation abundance at a spatial scale of 100 km x 100 km. LOVE estimates local vegetation abundance at the spatial scale of the relevant source area of pollen (RSAP sensu Sugita 1993) of the pollen site. REVEALS estimates are needed to apply LOVE in order to calculate the RSAP and the vegetation cover within the RSAP. The two models were validated theoretically and empirically. Two small bogs in southern Sweden were studied for pollen, plant macrofossil, charcoal, and coleoptera in order to reconstruct the local Holocene forest and fire history (e.g. Greisman and Gaillard 2009; Olsson et al. 2009). We applied the LOVE model in order to 1) compare the LOVE estimates with pollen percentages for a better understanding of the local forest history; 2) obtain more precise information on the local vegetation to explain between-sites differences in fire history. We used pollen records from two large lakes in Småland to obtain REVEALS estimates for twelve continuous 500-yrs time windows. Following the strategy of the Swedish VR LANDCLIM project (see Gaillard

Southern Annular Mode drives multicentury wildfire activity in southern South America.

PubMed

Holz, Andrés; Paritsis, Juan; Mundo, Ignacio A; Veblen, Thomas T; Kitzberger, Thomas; Williamson, Grant J; Aráoz, Ezequiel; Bustos-Schindler, Carlos; González, Mauro E; Grau, H Ricardo; Quezada, Juan M

2017-09-05

The Southern Annular Mode (SAM) is the main driver of climate variability at mid to high latitudes in the Southern Hemisphere, affecting wildfire activity, which in turn pollutes the air and contributes to human health problems and mortality, and potentially provides strong feedback to the climate system through emissions and land cover changes. Here we report the largest Southern Hemisphere network of annually resolved tree ring fire histories, consisting of 1,767 fire-scarred trees from 97 sites (from 22 °S to 54 °S) in southern South America (SAS), to quantify the coupling of SAM and regional wildfire variability using recently created multicentury proxy indices of SAM for the years 1531-2010 AD. We show that at interannual time scales, as well as at multidecadal time scales across 37-54 °S, latitudinal gradient elevated wildfire activity is synchronous with positive phases of the SAM over the years 1665-1995. Positive phases of the SAM are associated primarily with warm conditions in these biomass-rich forests, in which widespread fire activity depends on fuel desiccation. Climate modeling studies indicate that greenhouse gases will force SAM into its positive phase even if stratospheric ozone returns to normal levels, so that climate conditions conducive to widespread fire activity in SAS will continue throughout the 21st century.

Southern Annular Mode drives multicentury wildfire activity in southern South America

PubMed Central

Paritsis, Juan; Mundo, Ignacio A.; Veblen, Thomas T.; Kitzberger, Thomas; Williamson, Grant J.; Aráoz, Ezequiel; Bustos-Schindler, Carlos; González, Mauro E.; Grau, H. Ricardo; Quezada, Juan M.

2017-01-01

The Southern Annular Mode (SAM) is the main driver of climate variability at mid to high latitudes in the Southern Hemisphere, affecting wildfire activity, which in turn pollutes the air and contributes to human health problems and mortality, and potentially provides strong feedback to the climate system through emissions and land cover changes. Here we report the largest Southern Hemisphere network of annually resolved tree ring fire histories, consisting of 1,767 fire-scarred trees from 97 sites (from 22 °S to 54 °S) in southern South America (SAS), to quantify the coupling of SAM and regional wildfire variability using recently created multicentury proxy indices of SAM for the years 1531–2010 AD. We show that at interannual time scales, as well as at multidecadal time scales across 37–54 °S, latitudinal gradient elevated wildfire activity is synchronous with positive phases of the SAM over the years 1665–1995. Positive phases of the SAM are associated primarily with warm conditions in these biomass-rich forests, in which widespread fire activity depends on fuel desiccation. Climate modeling studies indicate that greenhouse gases will force SAM into its positive phase even if stratospheric ozone returns to normal levels, so that climate conditions conducive to widespread fire activity in SAS will continue throughout the 21st century. PMID:28827329

Southern Identity in "Southern Living" Magazine

ERIC Educational Resources Information Center

Lauder, Tracy

2012-01-01

A fantasy-theme analysis of the editors' letters in "Southern Living" magazine shows an editorial vision of valuing the past and showcasing unique regional qualities. In addition, a content analysis of the visual representation of race in the magazine's formative years and recent past validates that inhabitants of the region were portrayed…

Southern pulpwood production, 1973

Treesearch

Thomas R. Bellamy

1973-01-01

Southern pulpwood production in 1973 increased 6 percent to over 47.1 million cords--over 66 percent of the Nation's total. Plant byproducts used for pulping made the largest volume gain in history and now account for 27 percent of the total pulpwood production Southern pulpwood was processed at 126 mills, of which 111 are in the South. Pulping capacity in the 12...

Southern pulpwood production, 1992

Treesearch

Patrick E. Miller

1994-01-01

In 1992, southern pulpwood production increased 4 percent to 67.9 million cords. Roundwood production increased 1 percent to 47.3 million cords, and wood residue production increased 12 percent to 20.7 million cords. One pulpmill changed processes, reducing the number to 103. The pulping capacity of the 103 southern pulpmills was 133,400 tons per day. No new...

Southern pulpwood production, 1970

Treesearch

Roy C. Beltz

1971-01-01

Southern pulpwood production climbed to 42,152,410 cords in 1970, exceeding last year’s record by 3 percent. This increase is smaller than those of recent years. Pulping capacity also rose 3 percent to 83,316 tons per day, and four mills are currently under construction. Of the 119 mills processing southern pulpwood, 107 were located within the region.

The Southern Nevada Agency Partnership Science and Research Synthesis: Science to support land management in Southern Nevada

Treesearch

Jeanne C. Chambers; Matthew L. Brooks; Burton K. Pendleton; Carol B. Raish

2013-01-01

This synthesis provides information related to the Southern Nevada Agency Partnership (SNAP) Science and Research Strategy Goal 1 - to restore, sustain and enhance southern Nevada’s ecosystems - and Goal 2 - to provide for responsible use of southern Nevada’s lands in a manner that preserves heritage resources and promotes an understanding of human interaction with the...

Application of Organosilane Monolayer Template to Quantitative Evaluation of Cancer Cell Adhesive Ability

NASA Astrophysics Data System (ADS)

Tanii, Takashi; Sasaki, Kosuke; Ichisawa, Kota; Demura, Takanori; Beppu, Yuichi; Vu, Hoan Anh; Thanh Chi, Hoan; Yamamoto, Hideaki; Sato, Yuko

2011-06-01

The adhesive ability of two human pancreatic cancer cell lines was evaluated using organosilane monolayer templates (OMTs). Using the OMT, the spreading area of adhered cells can be limited, and this enables us to focus on the initial attachment process of adhesion. Moreover, it becomes possible to arrange the cells in an array and to quantitatively evaluate the number of attached cells. The adhesive ability of the cancer cells cultured on the OMT was controlled by adding (-)-epigallocatechin-3-gallate (EGCG), which blocks a receptor that mediates cell adhesion and is overexpressed in cancer cells. Measurement of the relative ability of the cancer cells to attach to the OMT revealed that the ability for attachment decreased with increasing EGCG concentration. The results agreed well with the western blot analysis, indicating that the OMT can potentially be employed to evaluate the adhesive ability of various cancer cells.

El Niño–Southern Oscillation diversity and Southern Africa teleconnections during Austral Summer

USGS Publications Warehouse

Hoell, Andrew; Funk, Christopher C.; Magadzire, Tamuka; Zinke, Jens; Husak, Gregory J.

2014-01-01

A wide range of sea surface temperature (SST) expressions have been observed during the El Niño–Southern Oscillation events of 1950–2010, which have occurred simultaneously with different global atmospheric circulations. This study examines the atmospheric circulation and precipitation during December–March 1950–2010 over the African Continent south of 15∘S, a region hereafter known as Southern Africa, associated with eight tropical Pacific SST expressions characteristic of El Niño and La Niña events. The self-organizing map method along with a statistical distinguishability test was used to isolate the SST expressions of El Niño and La Niña. The seasonal precipitation forcing over Southern Africa associated with the eight SST expressions was investigated in terms of the horizontal winds, moisture budget and vertical motion. El Niño events, with warm SST across the east and central Pacific Ocean and warmer than average SST over the Indian Ocean, are associated with precipitation reductions over Southern Africa. The regional precipitation reductions are forced primarily by large-scale mid-tropospheric subsidence associated with anticyclonic circulation in the upper troposphere. El Niño events with cooler than average SST over the Indian Ocean are associated with precipitation increases over Southern Africa associated with lower tropospheric cyclonic circulation and mid-tropospheric ascent. La Niña events, with cool SST anomalies over the central Pacific and warm SST over the west Pacific and Indian Ocean, are associated with precipitation increases over Southern Africa. The regional precipitation increases are forced primarily by lower tropospheric cyclonic circulation, resulting in mid-tropospheric ascent and an increased flux of moisture into the region.

Telomere erosion varies during in vitro aging of normal human fibroblasts from young and adult donors.

PubMed

Figueroa, R; Lindenmaier, H; Hergenhahn, M; Nielsen, K V; Boukamp, P

2000-06-01

The life span of normal fibroblasts in vitro (Hayflick limit) depends on donor age, and telomere shortening has been proposed as a potential mechanism. By quantitative fluorescence in situ hybridization and Southern blot analysis, we show progressive telomere loss to about 5 kb mean telomere restriction fragment length in fibroblasts from two adult donors within 40 population doublings, whereas in fibroblasts from two infant donors, telomere erosion is reduced, leaving a mean telomere restriction fragment length of approximately 7 kb at senescence (after approximately 60 population doublings). Aging of fibroblasts from both infant and adult donors was not accompanied by chromosomal abnormalities but was correlated with increased telomere repeat-binding factor 2 expression at both the protein and transcriptional level.

Clinical application of a rapid method using agarose gel electrophoresis and Western blotting to evaluate von Willebrand factor protease activity.

PubMed

Kirzek, D M; Rick, M E

2001-03-01

A method for evaluating the activity of the von Willebrand factor (vWF) protease is described, and a clinical application is illustrated. The procedure utilizes gel electrophoresis, Western blotting, and luminographic detection methods to evaluate the distribution of vWF multimers before and after incubation of clinical samples under conditions that favor proteolysis by this enzyme. Physiologically, the high-molecular-weight multimers of vWF are cleaved by the vWF protease under conditions of high shear stress in parts of the arterial circulation; cleavage of vWF multimers is also observed after exposure of vWF to denaturing agents in vitro and thus can serve as a laboratory test for the activity of the protease. vWF protease activity is decreased or absent in patients with thrombotic thrombocytopenic purpura due to an inhibiting autoantibody, and this leads to high levels of noncleaved vWF and to life-threatening thrombosis, thrombocytopenia and anemia. The assay evaluates the activity of the protease by assessing the cleavage of vWF multimers after patient plasmas are incubated in vitro under denaturing conditions. With the use of these electrophoresis and Western blotting techniques, patient plasmas can be rapidly assessed for the activity of the vWF protease which may aid in the treatment strategy for these patients.

Reverse line blot hybridisation screening of Pseudallescheria/Scedosporium species in patients with cystic fibrosis.

PubMed

Lu, Q; van den Ende, A H G Gerrits; de Hoog, G S; Li, R; Accoceberry, I; Durand-Joly, I; Bouchara, J-P; Hernandez, F; Delhaes, L

2011-10-01

The PCR-RLB (reverse line blot hybridisation) was applied as a molecular technique for the detection of members of Pseudallescheria and Scedosporium from sputum of patients with cystic fibrosis (CF). Fifty-nine sputum samples were collected from 52 CF patients, which were analysed by culture and PCR-RLB. Conventional and semi-selective culture yielded five positive samples, but the PCR-RLB hybridisation assay permitted the detection of members of Pseudallescheria/Scedosporium in 32 out of 52 patients (61.5%). In total, PCR-RLB yielded 47 positives. Pseudallescheria apiosperma was detected in 20 samples, while Pseudallescheria boydii and Pseudallescheria aurantiacum were detected in 17 and eight samples, respectively. Six samples gave a positive reaction with two distinct species-specific probes and one sample with three probes. In conclusion, the PCR-RLB assay described in this study allows the detection of Scedosporium spp. in CF sputum samples and the identification of Pseudallescheria apiosperma, P. boydii, S. aurantiacum, Scedosporium prolificans and Pseudallescheria minutispora. © 2011 Blackwell Verlag GmbH.

Analyzing the Factors That Can Limit the Acceptability to Introduce New Specializations in Higher Education Institutions: A Case Study of Higher Education Institutions of Southern Punjab, Pakistan

ERIC Educational Resources Information Center

Abbas, Wasim; Ahmed, Munir; Khalid, Rizwan; Yasmeen, Tehreem

2017-01-01

Purpose: The purpose of this paper is to provide a comprehensive view of barriers that restrict the adoption of new specializations and courses in the higher education institutions of Southern Punjab, Pakistan. Design/methodology/approach: Quantitative methods were used to get the responses from the respondents of different institution of target…

The Southern Hemisphere VLBI experiment

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

Preston, R.A.; Meier, D.L.; Louie, A.P.

1989-07-01

Six radio telescopes were operated as the first Southern Hemisphere VLBI array in April and May 1982. Observations were made at 2.3 and 8.4 GHz. This array provided VLBI modeling and hybrid imaging of celestial radio sources in the Southern Hemisphere, high-accuracy VLBI geodesy between Southern Hemisphere sites, and subarcsecond radio astrometry of celestial sources south of declination -45 deg. The goals and implementation of the array are discussed, the methods of modeling and hybrid image production are explained, and the VLBI structure of the sources that were observed is summarized. 36 refs.

Hepatitis E virus co-infection in HIV-infected patients in Foggia and Naples in southern Italy.

PubMed

Scotto, Gaetano; Grisorio, Benvenuto; Filippini, Pietro; Ferrara, Sergio; Massa, Salvatore; Bulla, Fabio; Martini, Salvatore; Filippini, Alberico; Tartaglia, Alessandra; Lo Muzio, Lorenzo; Fazio, Vincenzina

2015-01-01

Hepatitis E virus (HEV) infection represents an emerging infection in developed countries and is thought to be a zoonotic infection. It has recently been described as a new causative agent of acute and chronic hepatitis in immunosuppressed subjects, including HIV-infected patients. The aim of this study was to assess the sero-virological prevalence of HEV in HIV patients and in the general population as control group. A prospective and observational cohort study was carried out in two hospitals in southern Italy. The seroprevalence of HEV was determined in a cohort of 959 subjects, 509 (53%) of whom were HIV-positive patients and 450 were from the general population. Serum samples were tested for anti-HEV antibodies; repeatedly positive results were confirmed by a Western blot assay. In positive patients HEV RNA and genotypes were also determined. A total of 46 (4.8%) of the 959 serum samples examined were reactive to anti-HEV Ig and confirmed by Western blotting. The prevalence of HEV antibodies (IgG and/or IgM) was 2.7% in the control group and 6.7% in HIV-infected patients. Anti-HEV IgM was found in 6/46 (13.0%) of the anti-HEV Ig-positive serum samples, in 5/34 HIV patients and in 1/12 of the general population. No HIV-infected patient presented chronic hepatitis with HEV infection alone. This study indicates a higher circulation of HEV in HIV-infected patients, whereas a low prevalence of HEV antibodies in the general Italian population was shown. Chronic hepatitis with HEV alone was absent, while it was present in subjects with HIV-HEV, co-infected with hepatitis B virus (HBV) and/or hepatitis C virus (HCV).

Male specific genes from dioecious white campion identified by fluorescent differential display.

PubMed

Scutt, Charles P; Jenkins, Tom; Furuya, Masaki; Gilmartin, Philip M

2002-05-01

Fluorescent differential display (FDD) has been used to screen for cDNAs that are differentially up-regulated in male flowers of the dioecious plant Silene latifolia in which an X/Y chromosome system of sex determination operates. To adapt FDD to the cloning of large numbers of differential cDNAs, a novel method of confirming the differential expression of these has been devised. FDD gels were Southern electro-blotted and probed with mixtures of individual cDNA clones derived from different FDD product ligation reactions. These Southern blots were then stripped and re-probed with further mixtures of individual cloned FDD products to identify the maximum number of recombinant clones carrying the true differential amplification products. Of 135 differential bands identified by FDD, 56 differential amplification products were confirmed; these represent 23 unique differentially expressed genes as determined by virtual Northern analysis and two genes expressed at or below the level of detection by virtual Northern analysis. These two low expressed genes show bands of hybridization on genomic Southern blots that are specific to male plants, indicating that they are derived from, or closely related to, Y chromosome genes.

Silvicultural Considerations in Managing Southern Pine Stands in the Context of Southern Pine Beetle

Treesearch

James M. Guldin

2011-01-01

Roughly 30 percent of the 200 million acres of forest land in the South supports stands dominated by southern pines. These are among the most productive forests in the nation. Adapted to disturbance, southern pines are relatively easy to manage with even-aged methods such as clearcutting and planting, or the seed tree and shelterwood methods with natural regeneration....

Triton Southern Hemisphere

NASA Image and Video Library

1998-06-08

This polar projection from NASA Voyager 2 of Triton southern hemisphere provides a view of the southern polar cap and bright equatorial fringe. The margin of the cap is scalloped and ranges in latitude from +10 degrees to -30 degrees. The bright fringe is closely associated with the cap's margin; from it, diffuse bright rays extend north-northeast for hundreds of kilometers. The bright fringe probably consists of very fresh nitrogen frost or snow, and the rays consist of bright-fringe materials that were redistributed by north-moving Coriolis-deflected winds. http://photojournal.jpl.nasa.gov/catalog/PIA00423

Genetic basis of climatic adaptation in scots pine by bayesian quantitative trait locus analysis.

PubMed Central

Hurme, P; Sillanpää, M J; Arjas, E; Repo, T; Savolainen, O

2000-01-01

We examined the genetic basis of large adaptive differences in timing of bud set and frost hardiness between natural populations of Scots pine. As a mapping population, we considered an "open-pollinated backcross" progeny by collecting seeds of a single F(1) tree (cross between trees from southern and northern Finland) growing in southern Finland. Due to the special features of the design (no marker information available on grandparents or the father), we applied a Bayesian quantitative trait locus (QTL) mapping method developed previously for outcrossed offspring. We found four potential QTL for timing of bud set and seven for frost hardiness. Bayesian analyses detected more QTL than ANOVA for frost hardiness, but the opposite was true for bud set. These QTL included alleles with rather large effects, and additionally smaller QTL were supported. The largest QTL for bud set date accounted for about a fourth of the mean difference between populations. Thus, natural selection during adaptation has resulted in selection of at least some alleles of rather large effect. PMID:11063704

Southern Ocean Iron Experiment (SOFex)

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

Coale, Kenneth H.

The Southern Ocean Iron Experiment (SOFeX) was an experiment decades in the planning. It's implementation was among the most complex ship operations that SIO has been involved in. The SOFeX field expedition was successful in creating and tracking two experimentally enriched areas of the Southern Ocean, one characterized by low silicic acid, one characterized by high silicic acid. Both experimental sites were replete with abundant nitrate. About 100 scientists were involved overall. The major findings of this study were significant in several ways: (1) The productivity of the southern ocean is limited by iron availability. (2) Carbon uptake and fluxmore » is therefore controlled by iron availability (3) In spite of low silicic acid, iron promotes non-silicious phytoplankton growth and the uptake of carbon dioxide. (4) The transport of fixed carbon from the surface layers proceeds with a C:N ratio that would indicate differential remineralization of nitrogen at shallow depths. (5) These finding have major implications for modeling of carbon export based on nitrate utilization. (6) The general results of the experiment indicate that, beyond other southern ocean enrichment experiments, iron inputs have a much wider impact of productivity and carbon cycling than previously demonstrated. Scientific presentations: Coale, K., Johnson, K, Buesseler, K., 2002. The SOFeX Group. Eos. Trans. AGU 83(47) OS11A-0199. Coale, K., Johnson, K. Buesseler, K., 2002. SOFeX: Southern Ocean Iron Experiments. Overview and Experimental Design. Eos. Trans. AGU 83 (47) OS22D-01. Buesseler, K.,et al. 2002. Does Iron Fertilization Enhance Carbon Sequestration? Particle flux results from the Southern Ocean Iron Experiment. Eos. Trans. AGU 83 (47), OS22D-09. Johnson, K. et al. 2002. Open Ocean Iron Fertilization Experiments From IronEx-I through SOFeX: What We Know and What We Still Need to Understand. Eos. Trans. AGU 83 (47), OS22D-12. Coale, K. H., 2003. Carbon and Nutrient Cycling During the

[Western Blot diagnostic yield for simultaneous antibody-detection in patients with human cysticercosis, hydatidosis, and human fascioliasis].

PubMed

Davelois, Kelly; Escalante, Hermes; Jara, César

2016-01-01

. To determine the diagnostic yield using western blotting to simultaneously detect antibodies in patients with human cysticercosis, hydatidosis, and human fascioliasis. Materials and methods . Cross-sectional study of diagnostic yield assessment. Excretory/secretory antigens were obtained from Taenia solium larvae, Echinococcus granulosus cysts, and the adult flukes of Fasciola hepática, which were then separated using the polyacrylamide gel electrophoresis technique, transferred, and attached to a nitrocellulose membrane to be probed with sera from the patient infected with the three parasites. The sensitivity of the technique was assessed using 300 individual serum samples, 60 pools of two parasites, and 20 pools of three parasites with 75 sera from patients with other parasites, 10 from patients with other diseases, and 15 from patients without parasites. Results . The technique revealed 13 glycoproteins (GP): GP 35, 31, 24, 23, 18, 17, 14, and 13 kDa for cysticercosis; GP 8, 16, and 21 kDa for hydatidosis; and GP 17 and 23 kDa for fascioliasis. The test detected the presence of antibodies with a sensitivity of 96% (95% confidence interval [CI] = 94.62-98.54%) in the detection of one or the thirteen bands, a specificity of 100% (95% CI = 99.50-100.00%); individually, there was a sensitivity for cysticercosis of 97% (95% CI = 93.16-100.00%), for hydatidosis of 94% (95% CI = 88.85-99.15%) and for fascioliasis of 96% (95% CI = 91.66-100.00%). Conclusions . Western blotting is effective in the simultaneous detection of antibodies in patients with human cysticercosis, hydatidosis, and fascioliasis, and it can be used as a diagnostic test to either rule out or confirm the presence of antibodies in endemic areas.

Macroeconomics, (Adult) Education, and Poverty Eradication in Southern Africa

NASA Astrophysics Data System (ADS)

Nhamo, Senia; Nhamo, Godwell

2006-05-01

The Millennium Summit held in New York in September 2000 outlined the Millennium Development Goals (MDGs). The first of these involves the eradication of extreme poverty and hunger, setting two targets: halving by 2015 the percentage of the world's populace in 1990 with income less than US-1 a day (i.e., cutting this percentage from 27.9 to 14%); and halving the share of people who suffer from hunger. As for education, the MDGs seek to ensure that all children can complete primary schooling by 2015. Drawing on examples from selected southern African countries, the present study examines the need to strengthen economic support for (adult) education as an instrument of poverty eradication. It argues that human capital is one of the fundamental determinants of economic growth, and that this economic resource is essentially determined in both qualitative and quantitative regards by education.

Guidelines for regenerating southern pine beetle spots

Treesearch

J.C.G. Goelz; B.L. Strom; J.P. Barnett; M.A. Sword Sayer

2012-01-01

Southern pine forests are of exceptional commercial and ecological importance to the United States, and the southern pine beetle is their most serious insect pest. The southern pine beetle generally kills overstory pines, causing spots of tree mortality that are unpredictable in time and space and frequently disruptive to management activities and goals. The canopy...

Southern Coal Corporation Clean Water Settlement

EPA Pesticide Factsheets

Southern Coal Corporation is a coal mining and processing company headquartered in Roanoke, VA. Southern Coal Corporation and the following 26 affiliated entities are located in Alabama, Kentucky, Tennessee, Virginia and West Virginia

Studies in Site Evaluation for Southern Hardwoods

Treesearch

Thomas C. Nelson; William R. Beaufait

1956-01-01

SOUTHERN HARDWOODS, which supply more than half of the nation's hardwood lumber, are a complex and varied mixture of nearly 40 commercial species. On a good share of the 112 million acres of southern hardwood types the hardwoods are mixed with or have replaced southern pines. Faced with this profusion of species, the silviculturist's first problem is to know...

Quantitative characterization of nanoparticle agglomeration within biological media

NASA Astrophysics Data System (ADS)

Hondow, Nicole; Brydson, Rik; Wang, Peiyi; Holton, Mark D.; Brown, M. Rowan; Rees, Paul; Summers, Huw D.; Brown, Andy

2012-07-01

Quantitative analysis of nanoparticle dispersion state within biological media is essential to understanding cellular uptake and the roles of diffusion, sedimentation, and endocytosis in determining nanoparticle dose. The dispersion of polymer-coated CdTe/ZnS quantum dots in water and cell growth medium with and without fetal bovine serum was analyzed by transmission electron microscopy (TEM) and dynamic light scattering (DLS) techniques. Characterization by TEM of samples prepared by plunge freezing the blotted solutions into liquid ethane was sensitive to the dispersion state of the quantum dots and enabled measurement of agglomerate size distributions even in the presence of serum proteins where DLS failed. In addition, TEM showed a reduced packing fraction of quantum dots per agglomerate when dispersed in biological media and serum compared to just water, highlighting the effect of interactions between the media, serum proteins, and the quantum dots. The identification of a heterogeneous distribution of quantum dots and quantum dot agglomerates in cell growth medium and serum by TEM will enable correlation with the previously reported optical metrology of in vitro cellular uptake of this quantum dot dispersion. In this paper, we present a comparative study of TEM and DLS and show that plunge-freeze TEM provides a robust assessment of nanoparticle agglomeration state.

Dimensional stabilization of southern pines

Treesearch

E.T. Choong; H.M. Barnes

1969-01-01

The effectiveness of five dimensional stabilizing agents and three impregnation methods on southern pine was determined. Four southern pine species were studies in order to determine the effect of wood factors. The best dimensional stability was obtained when the wood was preswollen and the chemical was impregnated by a diffusion process. In general, polyethylene...

Southern Pinwheel

NASA Image and Video Library

2010-06-28

This image from NASA Wide-field Infrared Survey Explorer shows the nearby galaxy M83. This is a spiral galaxy approximately 15 million light-years away in the constellation Hydra, sometimes referred to as the southern Pinwheel galaxy.

Using in situ hybridization and PFGE Southern hybridization to detect translocation breakpoints in a BOR/TRPS patient cell line

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

Gu, J.Z.; Sapru, M.; Smith, D.

Branchio-oto-renal syndrome (BOR) is an autosomal dominant disorder characterized by ear malformations, cervical fistulae, hearing loss and renal abnormalities. We have integrated the Genethon YAC contig maps with additional markers in the chromosome 8q region genetically linked by a unique patient cell line. This cell line is from a patient who has both the branchio-oto-renal syndrome and tricho-rhino-phalangeal syndrome (TRPS). High resolution cytogenetics demonstrated a direct insertion of materials from 8q13.3q21.13 to 8q24.11. TRPS has been previously linked to deletions involving 8q24.11-q24.13. The rearrangement in this patient suggests that TRPS results from loss of gene function due to insertion atmore » the 8q24.11 breakpoint and the possible location for the BOR gene is at either of the two breakpoints of 8q13.3 and 8q21.13. We have constructed cosmid contigs in 8q24.11. In situ hybridization with cosmids mapped to these locations as probes has helped to narrow down the breakpoints. Combinations of cosmids on either side or overlapping the 8q24.11 breakpoint show split signals on one chromosome 8q arm due to insertion of the materials from the proximal region. Cosmids mapped to the TRPS deletion region have been used to hybridize to pulsed field gel genomic blots of DNA from the patient cell line and detected rearranged genomic fragments. Both in situ hybridization and genomic PFGE Southern blot will be used to precisely locate the breakpoints.« less

Hydrogeology and potential for ground-water development, carbonate-rock aquifers in southern Nevada and southeastern California

USGS Publications Warehouse

Burbey, T.J.

1997-01-01

Seventeen hydrographic areas in southern Nevada were assessed for the ground-water development potential of the underlying carbonate-rock aquifers on the basis of geologic and hydrologic information developed as part of the Nevada Carbonate Aquifers Study and information compiled from previous investigations. All selected areas lie within a miogeoclinal belt where thick accumulations of carbonate rock followed by major episodes of compression and extension have greatly modified the region. Most of the selected hydrographic areas lie within the less extended terranes; however, several areas, or parts of areas, lie within severely extended terranes where carbonate rocks have been greatly thinned, or where deformed blocks of carbonate rock are discontinuous and isolated from surrounding carbonate rock aquifers. Three principal criteria were used to assess the development potential of each selected hydrographic area. These quantitative criteria are: (1) depth to water, (2) depth to and thickness of carbonate rocks, and (3) water quality. Other site-specific factors, such as accessibility and potential effects of ground-water development, are also discussed. Results suggest that sites with high potential for development may be scarce in southern Nevada. Many areas described as favorable on the basis of the three quantitative criteria were deemed unfavorable on the basis of possible short- and long-term effects associated with development and on the amount of available data used to make the assessment. The most favorable sites may be in more severely extended terranes, where development of isolated blocks (of carbonate-rock aquifer material) would be less likely to affect neighboring areas.

A research design for the quantification of the neuropeptides substance p and calcitonin gene-related Peptide in rat skin using Western blot analysis.

PubMed

Lapin, Guilherme Abbud Franco; Hochman, Bernardo; Nishioka, Michele Akemi; Maximino, Jessica Ruivo; Chadi, Gerson; Ferreira, Lydia Masako

2015-06-01

To describe and standardize a protocol that overcomes the technical limitations of Western blot (WB) analysis in the quantification of the neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) following nociceptive stimuli in rat skin. Male Wistar rats (Rattus norvegicus albinus) weighing 250 to 350 g were used in this study. Elements of WB analysis were adapted by using specific manipulation of samples, repeated cycles of freezing and thawing, more thorough maceration, and a more potent homogenizer; increasing lytic reagents; promoting greater inhibition of protease activity; and using polyvinylidene fluoride membranes as transfer means for skin-specific protein. Other changes were also made to adapt the WB analysis to a rat model. University research center. Western blot analysis adapted to a rat model. This research design has proven effective in collecting and preparing skin samples to quantify SP and CGRP using WB analysis in rat skin. This study described a research design that uses WB analysis as a reproducible, technically accessible, and cost-effective method for the quantification of SP and CGRP in rat skin that overcomes technical biases.

Western blot analysis of sera from dogs with suspected food allergy.

PubMed

Favrot, Claude; Linek, Monika; Fontaine, Jacques; Beco, Luc; Rostaher, Ana; Fischer, Nina; Couturier, Nicolas; Jacquenet, Sandrine; Bihain, Bernard E

2017-04-01

Food allergy is often suspected in dogs with clinical signs of atopic dermatitis. This diagnosis is confirmed with an elimination diet and a subsequent challenge with regular food. Laboratory tests for the diagnosis of food allergy in dogs are unreliable and/or technically difficult. Cyno-DIAL ® is a Western blot method that might assist with the selection of an appropriate elimination diet. To evaluate the performance of Cyno-DIAL ® for the selection of an elimination diet and diagnosis of food allergy. Thirty eight dogs with atopic dermatitis completed an elimination diet. Combining the results of the diet trials and the challenges, 14 dogs were classified as food allergic (FA), 22 as nonfood-allergic and two as ambiguous cases. Amongst all dogs and amongst dogs with a clinical diagnosis of FA, 3% and 7% (respectively) were positive to Royal Canin Anallergenic ® , Vet-Concept Kanguru ® or Vet-Concept Dog Sana ® ; 8% and 7% to Hill's d/d Duck and Rice ® ; 8% and 21% to Hill's z/d Ultra Allergen Free ® ; 53% and 64% to Eukanuba Dermatosis FP ® ; and 32% and 43% to a home-cooked diet of horse meat, potatoes and zucchini. The specificity and sensitivity of Cyno-DIAL ® for diagnosing food allergy were 73% and 71%, respectively. Although Cyno-DIAL ® was considered potentially useful for identifying appropriate foods for elimination diet trials, it cannot be recommended for the diagnosis of food allergy. The Cyno-DIAL ® test performed better than some previously evaluated ELISA-based tests. © 2017 ESVD and ACVD.

Southern Pine Beetle Field Survey

Treesearch

Saul D. Petty

2011-01-01

Southern pine beetle (SPB) is one of the most formidable insect pests impacting southern forests. Federal, State, and private forest managers have always dealt with this pest in some capacity. One of the primary requirements for controlling SPB is locating infestations on the ground. Once the infestation has been located, data is collected and used in management...

Fusiform Rust of Southern Pines

Treesearch

W. R. Phelps; F. L. Czabator

1978-01-01

Fusiform rust, caused by the fungus Cronartium fusiforme Hedg. & Hunt ex Cumm., is distributed in the Southern United States from Maryland to Florida and west to Texas and southern Arkansas. Infections by the fungus, which develops at or near the point of infection, result in tapered, spindle-shaped swells, called galls, on branches and stems of pines. (see photo...

The southern stratospheric gravity wave hot spot: individual waves and their momentum fluxes measured by COSMIC GPS-RO

NASA Astrophysics Data System (ADS)

Hindley, N. P.; Wright, C. J.; Smith, N. D.; Mitchell, N. J.

2015-07-01

Nearly all general circulation models significantly fail to reproduce the observed behaviour of the southern wintertime polar vortex. It has been suggested that these biases result from an underestimation of gravity wave drag on the atmosphere at latitudes near 60° S, especially around the "hot spot" of intense gravity wave fluxes above the mountainous Southern Andes and Antarctic peninsula. Here, we use Global Positioning System radio occultation (GPS-RO) data from the COSMIC satellite constellation to determine the properties of gravity waves in the hot spot and beyond. We show considerable southward propagation to latitudes near 60° S of waves apparently generated over the southern Andes. We propose that this propagation may account for much of the wave drag missing from the models. Furthermore, there is a long leeward region of increased gravity wave energy that sweeps eastwards from the mountains over the Southern Ocean. Despite its striking nature, the source of this region has historically proved difficult to determine. Our observations suggest that this region includes both waves generated locally and orographic waves advected downwind from the hot spot. We describe and use a new wavelet-based analysis technique for the quantitative identification of individual waves from COSMIC temperature profiles. This analysis reveals different geographical regimes of wave amplitude and short-timescale variability in the wave field over the Southern Ocean. Finally, we use the increased numbers of closely spaced pairs of profiles from the deployment phase of the COSMIC constellation in 2006 to make estimates of gravity wave horizontal wavelengths. We show that, given sufficient observations, GPS-RO can produce physically reasonable estimates of stratospheric gravity wave momentum flux in the hot spot that are consistent with measurements made by other techniques. We discuss our results in the context of previous satellite and modelling studies and explain how they

Quantitative genetics

USDA-ARS?s Scientific Manuscript database

The majority of economically important traits targeted for cotton improvement are quantitatively inherited. In this chapter, the current state of cotton quantitative genetics is described and separated into four components. These components include: 1) traditional quantitative inheritance analysis, ...

The Southern Nevada Agency Partnership Science and Research Synthesis: Science to support land management in Southern Nevada - Executive Summary

Treesearch

Jeanne C. Chambers; Matthew L. Brooks; Burton K. Pendleton; Carol B. Raish

2013-01-01

This synthesis provides information related to the Southern Nevada Agency Partnership (SNAP) Science and Research Strategy Goal 1 - to restore, sustain and enhance southern Nevada’s ecosystems - and Goal 2 - to provide for responsible use of southern Nevada’s lands in a manner that preserves heritage resources and promotes an understanding of human interaction with the...

Southern Ocean warming due to human influence

NASA Astrophysics Data System (ADS)

Fyfe, John C.

2006-10-01

I show that the latest series of climate models reproduce the observed mid-depth Southern Ocean warming since the 1950s if they include time-varying changes in anthropogenic greenhouse gases, sulphate aerosols and volcanic aerosols in the Earth's atmosphere. The remarkable agreement between observations and state-of-the art climate models suggests significant human influence on Southern Ocean temperatures. I also show that climate models that do not include volcanic aerosols produce mid-depth Southern Ocean warming that is nearly double that produced by climate models that do include volcanic aerosols. This implies that the full effect of human-induced warming of the Southern Ocean may yet to be realized.

A New Method for Quantitative Immunoblotting of Endogenous α-Synuclein

PubMed Central

Newman, Andrew J.; Selkoe, Dennis; Dettmer, Ulf

2013-01-01

β-Sheet-rich aggregates of α-synuclein (αSyn) are the hallmark neuropathology of Parkinson’s disease and related synucleinopathies, whereas the principal native structure of αSyn in healthy cells - unfolded monomer or α-helically folded oligomer - is under debate. Our recent crosslinking analysis of αSyn in intact cells showed that a large portion of endogenous αSyn can be trapped as oligomers, most notably as apparent tetramers. One challenge in such studies is accurately quantifying αSyn Western blot signals among samples, as crosslinked αSyn trends toward increased immunoreactivity. Here, we analyzed this phenomenon in detail and found that treatment with the reducible amine-reactive crosslinker DSP strongly increased αSyn immunoreactivity even after cleavage with the reducing agent β-mercaptoethanol. The effect was observed with all αSyn antibodies tested and in all sample types from human brain homogenates to untransfected neuroblastoma cells, permitting easy detection of endogenous αSyn in the latter, which had long been considered impossible. Coomassie staining of blots before and after several hours of washing revealed complete retention of αSyn after DSP/β-mercaptoethanol treatment, in contrast to a marked loss of αSyn without this treatment. The treatment also enhanced immunodetection of the homologs β- and γ-synuclein and of histones, another group of small, lysine-rich proteins. We conclude that by neutralizing positive charges and increasing protein hydrophobicity, amine crosslinker treatment promotes adhesion of αSyn to blotting membranes. These data help explain the recent report of fixing αSyn blots with paraformaldehyde after transfer, which we find produces similar but weaker effects. DSP/β-mercaptoethanol treatment of Western blots should be particularly useful to quantify low-abundance αSyn forms such as extracellular and post-translationally modified αSyn and splice variants. PMID:24278419

Southern California Daily Energy Report

EIA Publications

2016-01-01

EIA has updated its Southern California Daily Energy Report to provide additional information on key energy market indicators for the winter season. The dashboard includes information that EIA regularly compiles about energy operations and the management of natural gas and electricity systems in Southern California in the aftermath of a leak at the Aliso Canyon natural gas storage facility outside of Los Angeles

The Biogeochemical Role of Antarctic Krill and Baleen Whales in Southern Ocean Nutrient Cycling.

NASA Astrophysics Data System (ADS)

Ratnarajah, L.

2015-12-01

Iron limits primary productivity in large areas of the Southern Ocean. It has been suggested that baleen whales form a crucial part of biogeochemical cycling processes through the consumption of nutrient-rich krill and subsequent defecation, but evidence on their contribution is scarce. We analysed the concentration of iron in Antarctic krill and baleen whale faeces and muscle. Iron concentrations in Antarctic krill were over 1 million times higher, and whale faecal matter were almost 10 million times higher than typical Southern Ocean High Nutrient Low Chlorophyll seawater concentrations. This suggests that Antarctic krill act as a reservoir of in in Southern Ocean surface waters, and that baleen whales play an important role in converting this fixed iron into a liquid form in their faeces. We developed an exploratory model to examine potential contribution of blue, fin and humpback whales to the Southern Ocean iron cycle to explore the effect of the recovery of great whales to historical levels. Our results suggest that pre-exploitation populations of blue whales and, to a lesser extent fin and humpback whales, could have contributed to the more effective recycling of iron in surface waters, resulting in enhanced phytoplankton production. This enhanced primary productivity is estimated to be: 8.3 x 10-5 to 15 g C m-2 yr-1 (blue whales), 7 x 10-5 to 9 g C m-2 yr-1 (fin whales), and 10-5 to 1.7 g C m-2 yr-1 (humpback whales). To put these into perspective, current estimates of primary production in the Southern Ocean from remotely sensed ocean colour are in the order of 57 g C m-2 yr-1 (south of 50°). The high degree of uncertainty around the magnitude of these increases in primary productivity is mainly due to our limited quantitative understanding of key biogeochemical processes including iron content in krill, krill consumption rates by whales, persistence of iron in the photic zone, bioavailability of retained iron, and carbon-to-iron ratio of phytoplankton

Smoke from Fires in Southern Mexico

NASA Image and Video Library

2002-05-08

On May 2, 2002, numerous fires in southern Mexico sent smoke drifting northward over the Gulf of Mexico. These views from NASA Terra satellite illustrate the smoke extent over parts of the Gulf and the southern Mexican states of Tabasco, Campeche and Ch

Improved Protein Arrays for Quantitative Systems Analysis of the Dynamics of Signaling Pathway Interactions

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

Yang, Chin-Rang

Astronauts and workers in nuclear plants who repeatedly exposed to low doses of ionizing radiation (IR, <10 cGy) are likely to incur specific changes in signal transduction and gene expression in various tissues of their body. Remarkable advances in high throughput genomics and proteomics technologies enable researchers to broaden their focus from examining single gene/protein kinetics to better understanding global gene/protein expression profiling and biological pathway analyses, namely Systems Biology. An ultimate goal of systems biology is to develop dynamic mathematical models of interacting biological systems capable of simulating living systems in a computer. This Glue Grant is to complementmore » Dr. Boothman’s existing DOE grant (No. DE-FG02-06ER64186) entitled “The IGF1/IGF-1R-MAPK-Secretory Clusterin (sCLU) Pathway: Mediator of a Low Dose IR-Inducible Bystander Effect” to develop sensitive and quantitative proteomic technology that suitable for low dose radiobiology researches. An improved version of quantitative protein array platform utilizing linear Quantum dot signaling for systematically measuring protein levels and phosphorylation states for systems biology modeling is presented. The signals are amplified by a confocal laser Quantum dot scanner resulting in ~1000-fold more sensitivity than traditional Western blots and show the good linearity that is impossible for the signals of HRP-amplification. Therefore this improved protein array technology is suitable to detect weak responses of low dose radiation. Software is developed to facilitate the quantitative readout of signaling network activities. Kinetics of EGFRvIII mutant signaling was analyzed to quantify cross-talks between EGFR and other signaling pathways.« less

Development of Prevotella intermedia-specific PCR primers based on the nucleotide sequences of a DNA probe Pig27.

PubMed

Kim, Min Jung; Hwang, Kyung Hwan; Lee, Young-Seok; Park, Jae-Yoon; Kook, Joong-Ki

2011-03-01

The aim of this study was to develop Prevotella intermedia-specific PCR primers based on the P. intermedia-specific DNA probe. The P. intermedia-specific DNA probe was screened by inverted dot blot hybridization and confirmed by Southern blot hybridization. The nucleotide sequences of the species-specific DNA probes were determined using a chain termination method. Southern blot analysis showed that the DNA probe, Pig27, detected only the genomic DNA of P. intermedia strains. PCR showed that the PCR primers, Pin-F1/Pin-R1, had species-specificity for P. intermedia. The detection limits of the PCR primer sets were 0.4pg of the purified genomic DNA of P. intermedia ATCC 49046. These results suggest that the PCR primers, Pin-F1/Pin-R1, could be useful in the detection of P. intermedia as well as in the development of a PCR kit in epidemiological studies related to periodontal diseases. Crown Copyright © 2010. Published by Elsevier B.V. All rights reserved.

Tenacious Southern Progressives: Confounding Mencken's Myth of Mediocrity.

ERIC Educational Resources Information Center

Allen, Louise; Reynolds, Katherine

After H.L. Mencken in a 1920 essay labeled the American South "the Sahara of the Bozart," the journalist Gerald Johnson debated with him the merits of southern intellectual life primarily as indicated in southern literature. There were noteworthy southern artists, journalists, social anthropologists, and educators, ranging from the…

Estimating the number of double-strand breaks formed during meiosis from partial observation.

PubMed

Toyoizumi, Hiroshi; Tsubouchi, Hideo

2012-12-01

Analyzing the basic mechanism of DNA double-strand breaks (DSB) formation during meiosis is important for understanding sexual reproduction and genetic diversity. The location and amount of meiotic DSBs can be examined by using a common molecular biological technique called Southern blotting, but only a subset of the total DSBs can be observed; only DSB fragments still carrying the region recognized by a Southern blot probe are detected. With the assumption that DSB formation follows a nonhomogeneous Poisson process, we propose two estimators of the total number of DSBs on a chromosome: (1) an estimator based on the Nelson-Aalen estimator, and (2) an estimator based on a record value process. Further, we compared their asymptotic accuracy.

Development and application of a quantitative multiplexed small GTPase activity assay using targeted proteomics.

PubMed

Zhang, Cheng-Cheng; Li, Ru; Jiang, Honghui; Lin, Shujun; Rogalski, Jason C; Liu, Kate; Kast, Juergen

2015-02-06

Small GTPases are a family of key signaling molecules that are ubiquitously expressed in various types of cells. Their activity is often analyzed by western blot, which is limited by its multiplexing capability, the quality of isoform-specific antibodies, and the accuracy of quantification. To overcome these issues, a quantitative multiplexed small GTPase activity assay has been developed. Using four different binding domains, this assay allows the binding of up to 12 active small GTPase isoforms simultaneously in a single experiment. To accurately quantify the closely related small GTPase isoforms, a targeted proteomic approach, i.e., selected/multiple reaction monitoring, was developed, and its functionality and reproducibility were validated. This assay was successfully applied to human platelets and revealed time-resolved coactivation of multiple small GTPase isoforms in response to agonists and differential activation of these isoforms in response to inhibitor treatment. This widely applicable approach can be used for signaling pathway studies and inhibitor screening in many cellular systems.

Quantifying export production in the Southern Ocean: Implications for the Baxs proxy

NASA Astrophysics Data System (ADS)

Hernandez-Sanchez, Maria T.; Mills, Rachel A.; Planquette, HéLèNe; Pancost, Richard D.; Hepburn, Laura; Salter, Ian; Fitzgeorge-Balfour, Tania

2011-12-01

The water column and sedimentary Baxs distribution around the Crozet Plateau is used to decipher the controls and timing of barite formation and to evaluate how export production signals are recorded in sediments underlying a region of natural Fe fertilization within the Fe limited Southern Ocean. Export production estimated from preserved, vertical sedimentary Baxs accumulation rates are compared with published export fluxes assessed from an integrated study of the biological carbon pump to determine the validity of Baxs as a quantitative proxy under different Fe supply conditions typical of the Southern Ocean. Detailed assessment of the geochemical partitioning of Ba in sediments and the lithogenic end-member allows appropriate correction of the bulk Ba content and determination of the Baxs content of sediments and suspended particles. The upper water column distribution of Baxs is extremely heterogeneous spatially and temporally. Organic carbon/Baxs ratios in deep traps from the Fe fertilized region are similar to other oceanic settings allowing quantification of the inferred carbon export based on established algorithms. There appears to be some decoupling of POC and Ba export in the Fe limited region south of the Plateau. The export production across the Crozet Plateau inferred from the Baxs sedimentary proxy indicates that the Fe fertilized area to the north of the Plateau experiences enhanced export relative to equivalent Southern Ocean settings throughout the Holocene and that this influence may also have impacted the site to the south for significant periods. This interpretation is corroborated by alternative productivity proxies (opal accumulation, 231Paxs/230Thxs). Baxs can be used to quantify export production in complex settings such as naturally Fe-fertilized (volcanoclastic) areas, providing appropriate lithogenic correction is undertaken, and sediment focusing is corrected for along with evaluation of barite preservation.

Granulites from Northwest Indian Shield: Their differences and similarities with Southern Indian granulite terrain

NASA Technical Reports Server (NTRS)

Sharma, R. S.

1988-01-01

Granulite facies suite in the NW Indian Shield is exposed at Sand Mata, Udaipur district, Rajasthan, as an oval-shaped massif within amphibolite facies rocks of the Banded Gneissic Complex (3.5 to 2.6 b.y. old) - a possible analogue of the Peninsular gneiss of Dharwar craton. On the basis of quantitative P-T estimates, combined with the textural evidence for the crystallization sequence of the Al-silicate polymorphs (kyanite to sillimanite to kyanite) in the pelitic granulite, the deduced P-T path for the Sand Mata granulites is the reverse of that characterizing the Plate tectonic collision zone. It, however, agrees with the P-T path inferred in the case of the southern Indian granulitic rocks.

iTRAQ-based quantitative proteomics analysis of molecular mechanisms associated with Bombyx mori (Lepidoptera) larval midgut response to BmNPV in susceptible and near-isogenic strains.

PubMed

Yu, Haizhong; Wang, Xueyang; Xu, Jiaping; Ma, Yan; Zhang, Shangzhi; Yu, Dong; Fei, Dongqiong; Muhammad, Azharuddin

2017-08-08

Bombyx mori nucleopolyhedrovirus (BmNPV) has been identified as a major pathogen responsible for severe economic loss. Most silkworm strains are susceptible to BmNPV, with only a few highly resistant strains thus far identified. Here we investigated the molecular basis of silkworm resistance to BmNPV using susceptible (the recurrent parent P50) and resistant (near-isogenic line BC9) strains and a combination of iTRAQ-based quantitative proteomics, reverse-transcription quantitative PCR and Western blotting. By comparing the proteomes of infected and non-infected P50 and BC9 silkworms, we identified 793 differentially expressed proteins (DEPs). By gene ontology and KEGG enrichment analyses, we found that these DEPs are preferentially involved in metabolism, catalytic activity, amino sugar and nucleotide sugar metabolism and carbon metabolism. 114 (14.38%) DEPs were associated with the cytoskeleton, immune response, apoptosis, ubiquitination, translation, ion transport, endocytosis and endopeptidase activity. After removing the genetic background and individual immune stress response proteins, we identified 84 DEPs were found that are potentially involved in resistance to BmNPV. Further studies showed that a serine protease was down-regulated in P50 and up-regulated in BC9 after BmNPV infection. Taken together, these results provide insights into the molecular mechanism of silkworm response to BmNPV. Bombyx mori nucleopolyhedrovirus (BmNPV) is highly pathogenic, causing serious losses in sericulture every year. However, the molecular mechanisms of BmNPV infection and host defence remain unclear. Here we combined quantitative proteomic, bioinformatics, RT-qPCR and Western blotting analyses and found that BmNPV invasion causes complex protein alterations in the larval midgut, and that these changes are related to cytoskeleton, immune response, apoptosis, ubiquitination, translation, ion transport, endocytosis and endopeptidase activity. Five important differentially

Taeniasis-cysticercosis in Southern Ecuador: assessment of infection status using multiple laboratory diagnostic tools.

PubMed

Rodriguez-Hidalgo, R; Benitez-Ortiz, W; Praet, N; Saa, L R; Vercruysse, J; Brandt, J; Dorny, P

2006-11-01

Taenia solium-taeniasis and cysticercosis were studied in the human and porcine populations of a rural community in the Southern Ecuadorian Andes. From the 1059 inhabitants, 800 serum samples and 958 stool samples could be collected. In addition, 646 from the estimated 1148 pigs were tongue inspected. Circulating antigen was detected by enzyme linked immunosorbent assay (Ag-ELISA) in 2.25% of the human population, whereas intestinal taeniasis was detected in 1.46% by the formalin-ether technique. Following treatment and recovery of tapeworm fragments these were all identified as T. solium. Porcine cysticercosis was diagnosed in 3.56% of the pigs by tongue inspection. In addition, enzyme linked immunoelectrotransfer blot (EITB) was performed on a subset group of 100 humans to confirm the results of the Ag-ELISA. One hundred serum samples from pigs were also analysed by EITB. It appeared that 43 and 74% of humans and pigs had antibodies against T. solium cysticerci, respectively. It is concluded that contrary to the high exposure of the human population to T. solium that is suggested by EITB, the number of active cysticercosis cases, diagnosed by Ag-ELISA, was low, which may indicate endemic stability. The further use of complementary diagnostic methods for a better understanding of the epidemiology of T. solium is suggested.

Analysis of differentially expressed genes in two immunologically distinct strains of Eimeria maxima using suppression subtractive hybridization and dot-blot hybridization

PubMed Central

2014-01-01

Background It is well known that different Eimeria maxima strains exhibit significant antigenic variation. However, the genetic basis of these phenotypes remains unclear. Methods Total RNA and mRNA were isolated from unsporulated oocysts of E. maxima strains SH and NT, which were found to have significant differences in immunogenicity in our previous research. Two subtractive cDNA libraries were constructed using suppression subtractive hybridization (SSH) and specific genes were further analyzed by dot-blot hybridization and qRT-PCR analysis. Results A total of 561 clones were selected from both cDNA libraries and the length of the inserted fragments was 0.25–1.0 kb. Dot-blot hybridization revealed a total of 86 differentially expressed clones (63 from strain SH and 23 from strain NT). Nucleotide sequencing analysis of these clones revealed ten specific contigs (six from strain SH and four from strain NT). Further analysis found that six contigs from strain SH and three from strain NT shared significant identities with previously reported proteins, and one contig was presumed to be novel. The specific differentially expressed genes were finally verified by RT-PCR and qRT-PCR analyses. Conclusions The data presented here suggest that specific genes identified between the two strains may be important molecules in the immunogenicity of E. maxima that may present potential new drug targets or vaccine candidates for coccidiosis. PMID:24894832

A Comparison of Antibacterial Activity of Selected Thyme (Thymus) Species by Means of the Dot Blot Test with Direct Bioautographic Detection.

PubMed

Orłowska, Marta; Kowalska, Teresa; Sajewicz, Mieczysław; Jesionek, Wioleta; Choma, Irena M; Majer-Dziedzic, Barbara; Szymczak, Grażyna; Waksmundzka-Hajnos, Monika

2015-01-01

Bioautography carried out with the aid of thin-layer chromatographic adsorbents can be used to assess antibacterial activity in samples of different origin. It can either be used as a simple and cost-effective detection method applied to a developed chromatogram, or to the dot blot test performed on a chromatographic plate, where total antibacterial activity of a sample is scrutinized. It was an aim of this study to compare antibacterial activity of 18 thyme (Thymus) specimens and species (originating from the same gardening plot and harvested in the same period of time) by means of a dot blot test with direct bioautography. A two-step extraction of herbal material was applied, and at step two the polar fraction of secondary metabolites was obtained under the earlier optimized extraction conditions [methanol-water (27+73, v/v), 130°C]. This fraction was then tested for its antibacterial activity against Bacillus subtilis bacteria. It was established that all investigated extracts exhibited antibacterial activity, yet distinct differences were perceived in the size of the bacterial growth inhibition zones among the compared thyme species. Based on the results obtained, T. citriodorus "golden dwarf" (sample No. 5) and T. marschallianus (sample No. 6) were selected as promising targets for further investigations and possible inclusion in a herbal pharmacopeia, which is an essential scientific novelty of this study.

Reduced C9orf72 protein levels in frontal cortex of amyotrophic lateral sclerosis and frontotemporal degeneration brain with the C9ORF72 hexanucleotide repeat expansion.

PubMed

Waite, Adrian J; Bäumer, Dirk; East, Simon; Neal, James; Morris, Huw R; Ansorge, Olaf; Blake, Derek J

2014-07-01

An intronic G(4)C(2) hexanucleotide repeat expansion in C9ORF72 is a major cause of amyotrophic lateral sclerosis and frontotemporal lobar degeneration. Several mechanisms including RNA toxicity, repeat-associated non-AUG translation mediated dipeptide protein aggregates, and haploinsufficiency of C9orf72 have been implicated in the molecular pathogenesis of this disorder. The aims of this study were to compare the use of two different Southern blot probes for detection of repeat expansions in an amyotrophic lateral sclerosis and frontotemporal lobar degeneration pathological cohort and to determine the levels of C9orf72 transcript variants and protein isoforms in patients versus control subjects. Our Southern blot studies identified smaller repeat expansions (250-1800 bp) that were only detectable with the flanking probe highlighting the potential for divergent results using different Southern blotting protocols that could complicate genotype-phenotype correlation studies. Further, we characterize a new C9orf72 antibody and show for the first time decreased C9orf72 protein levels in the frontal cortex from patients with a pathological hexanucleotide repeat expansion. These data suggest that a reduction in C9orf72 protein may be a consequence of the disease. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

Confirmation of the immunoreactivity of monoclonal anti-human C-terminal EGFR antibodies in bronze Corydoras Corydoras aeneus (Callichthyidae Teleostei) by Western Blot method.

PubMed

Mytych, Jennifer; Satora, Leszek; Kozioł, Katarzyna

2018-02-01

Bronze corydoras (Corydoras aeneus) uses the distal part of the intestine as accessory respiratory organ. Our previous study showed the presence of epidermal growth factor receptor (EGFR) cytoplasmic domain in the digestive tract of the bronze corydoras. In this study, using Western Blot method, we validated the results presented in the previous research. In detail, results of Western Blot analysis on digestive and respiratory part of bronze corydoras intestine homogenates confirmed the immunoreactivity of anti-cytoplasmic domain (C-terminal) human EGFR antibodies with protein band of approximately 180kDa (EGFR molecular weight). This indicates a high homology of EGFR domain between these species and the possibility of such antibody use in bronze corydoras. Statistically significantly higher EGFR expression was observed in the respiratory part of intestine when compared to the digestive part. This implies higher proliferation activity and angiogenesis of epithelium in this part of intestine, creating conditions for air respiration. Therefore, Corydoras aeneus may be considered as a model organism for the molecular studies of the mechanisms of epithelial proliferation initiation and inhibition depending on hypoxia and normoxia. Copyright © 2017. Published by Elsevier GmbH.

2001-2002 Southern California Regional Travel Survey | Transportation

Science.gov Websites

Secure Data Center | NREL 01-2002 Southern California Regional Travel Survey 2001-2002 Southern California Regional Travel Survey The 2001-2002 Southern California Regional Travel Survey collected data on conduct the survey following the 2000 decennial census. Methodology Survey data collection occurred in

Qualitative and quantitative determination of enterobacterial common antigen (ECA) with monoclonal antibodies: expression of ECA by two Actinobacillus species.

PubMed Central

Böttger, E C; Jürs, M; Barrett, T; Wachsmuth, K; Metzger, S; Bitter-Suermann, D

1987-01-01

The presence and quantity of the enterobacterial common antigen (ECA) in several species belonging to the family Enterobacteriaceae as well as to other gram-negative families were determined by a solid-phase enzyme-linked immunosorbent assay system and Western blotting by using mouse monoclonal antibodies specific for ECA. Except for Erwinia chrysanthemi, previously known to be an exception, all species known or presumed to belong to Enterobacteriaceae produced ECA (89 of 90 species). Most species not belonging to Enterobacteriaceae did not produce ECA (25 of 28 species), with one already known (Plesiomonas shigelloides) and two hitherto unknown (Actinobacillus equuli and Actinobacillus suis) exceptions. Interestingly, all strains of P. shigelloides produced ECA, regardless of the presence of the Shigella sonnei cross-reacting O antigen. Quantitation of the amount of ECA in members of the family Enterobacteriaceae revealed a remarkable heterogeneity among genera and species as well as within one species. We conclude that the rapid, sensitive, and reliable determination of ECA is a useful aid in taxonomic classification and may help to characterize the relatedness of the family Enterobacteriaceae to other families. However, a quantitative analysis of ECA appears to be without value for these purposes. Images PMID:3818929

Comparison of survival patterns of northern and southern genotypes of the North American tick Ixodes scapularis (Acari: Ixodidae) under northern and southern conditions

USGS Publications Warehouse

Ginsberg, Howard S.; Rulison, Eric L.; Azevedo, Alexandra; Pang, Genevieve C.; Kuczaj, Isis M.; Tsao, Jean I.; LeBrun, Roger A.

2014-01-01

BackgroundSeveral investigators have reported genetic differences between northern and southern populations of Ixodes scapularis in North America, as well as differences in patterns of disease transmission. Ecological and behavioral correlates of these genetic differences, which might have implications for disease transmission, have not been reported. We compared survival of northern with that of southern genotypes under both northern and southern environmental conditions in laboratory trials.MethodsSubadult I. scapularis from laboratory colonies that originated from adults collected from deer from several sites in the northeastern, north central, and southern U.S. were exposed to controlled conditions in environmental chambers. Northern and southern genotypes were exposed to light:dark and temperature conditions of northern and southern sites with controlled relative humidities, and mortality through time was recorded.ResultsTicks from different geographical locations differed in survival patterns, with larvae from Wisconsin surviving longer than larvae from Massachusetts, South Carolina or Georgia, when held under the same conditions. In another experiment, larvae from Florida survived longer than larvae from Michigan. Therefore, survival patterns of regional genotypes did not follow a simple north–south gradient. The most consistent result was that larvae from all locations generally survived longer under northern conditions than under southern conditions.ConclusionsOur results suggest that conditions in southern North America are less hospitable than in the north to populations of I. scapularis. Southern conditions might have resulted in ecological or behavioral adaptations that contribute to the relative rarity of I. scapularis borne diseases, such as Lyme borreliosis, in the southern compared to the northern United States.

Southern deepwater swamps

Treesearch

William H. Conner

1998-01-01

The authors define, classify, and analyze the economic significance of southern deepwater swamps. They discuss the physical environment, vegetational communities, animal communities, management issues, and research needs for this complex resource.

ZyFISH: A Simple, Rapid and Reliable Zygosity Assay for Transgenic Mice

PubMed Central

McHugh, Donal; O’Connor, Tracy; Bremer, Juliane; Aguzzi, Adriano

2012-01-01

Microinjection of DNA constructs into fertilized mouse oocytes typically results in random transgene integration at a single genomic locus. The resulting transgenic founders can be used to establish hemizygous transgenic mouse lines. However, practical and experimental reasons often require that such lines be bred to homozygosity. Transgene zygosity can be determined by progeny testing assays which are expensive and time-consuming, by quantitative Southern blotting which is labor-intensive, or by quantitative PCR (qPCR) which requires transgene-specific design. Here, we describe a zygosity assessment procedure based on fluorescent in situ hybridization (zyFISH). The zyFISH protocol entails the detection of transgenic loci by FISH and the concomitant assignment of homozygosity using a concise and unbiased scoring system. The method requires small volumes of blood, is scalable to at least 40 determinations per assay, and produces results entirely consistent with the progeny testing assay. This combination of reliability, simplicity and cost-effectiveness makes zyFISH a method of choice for transgenic mouse zygosity determinations. PMID:22666404

Straight studs are produced from southern pine cordwood

Treesearch

Peter Koch

1967-01-01

A Process for converting southern pine veneer cores into 8-foot 2 by 4's of SPIB Stud grade and better has been developed at the Alexandria, Louisiana, Utilization Laboratory of the Southern Forest Experiment Station. The research leading to this development suggests that a similiar process would be practical for converting 8-foot southern pine cordwood into studs...

Southern Forest Resource Assessment - Technical Report

Treesearch

David N. Wear; John G. Greis

2002-01-01

The southern forest resource assessment provides a comprehensive analysis of the history, status, and likely future of forests in the Southern United States. Twenty-three chapters address questions regarding social/economic systems, terrestrial ecosystems, water and aquatic ecosystems, forest health, and timber management; 2 additional chapters provide a background on...

The Distribution of Southern Black Population.

ERIC Educational Resources Information Center

Tucker, Jack; Lee, Everett S.

Between 1960 and 1970, the black population of the southern United States increased only four percent by comparison with a national increase in black population of twenty percent. Considering the many factors involved in population redistribution, this report compares southern and overall national trends in black population and analyzes the…

DIET OF THE SOUTHERN TOAD FROM THE SOUTHERN EVERGLADES

EPA Science Inventory

We examined the diet of a February-May sample of the southern toad (Bufo Terrestris) from the Everglades National Park. Above the familial level, 13 taxa were consumed, but ants (Hymenoptera) and beetles (Coleoptera) were consumed most by, and in the greatest number of s...

Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae.

PubMed

Wang, Hye-young; Kim, Hyunjung; Kim, Yeun; Bang, Hyeeun; Kim, Jong-Pill; Hwang, Joo Hwan; Cho, Sang-Nae; Kim, Tae Ue; Lee, Hyeyoung

2015-10-01

Drug resistance in Mycobacterium leprae is a significant problem in countries where leprosy is endemic. A sensitive, specific, and high-throughput reverse blot hybridization assay (REBA) for the detection of genotypic resistance to rifampicin (RIF) was designed and evaluated. It has been shown that resistance to RIF in M. leprae involves mutations in the rpoB gene encoding the -subunit of the RNA polymerase. The PCR-REBA simultaneously detects both 6 wild-type regions and 5 different mutations (507 AGC, 513 GTG, 516 TAT, 531 ATG, and 531 TTC) including the most prevalent mutations at positions 507 and 531. Thirty-one clinical isolates provided by Korea Institute of Hansen-s Disease were analyzed by PCR-REBA with RIF resistance of rpoB gene. As a result, missense mutations at codons 507 AGC and 531 ATG with 2-nucleotide substitutions were found in one sample, and a missense mutation at codon 516 TAT and ΔWT6 (deletion of 530-534) was found in another sample. These cases were confirmed by DNA sequence analysis. This rapid, simple, and highly sensitive assay provides a practical alternative to sequencing for genotypic evaluation of RIF resistance in M. leprae.

Identification of Yeast V-ATPase Mutants by Western Blots Analysis of Whole Cell Lysates

NASA Astrophysics Data System (ADS)

Parra-Belky, Karlett

2002-11-01

A biochemistry laboratory was designed for an undergraduate course to help students better understand the link between molecular engineering and biochemistry. Students identified unknown yeast strains with high specificity using SDS-PAGE and Western blot analysis of whole cell lysates. This problem-solving exercise is a common application of biochemistry in biotechnology research. Three different strains were used: a wild-type and two mutants for the proton pump vacuolar ATPase (V-ATPase). V-ATPases are multisubunit enzymes and the mutants used were deletion mutants; each lacked one structural gene of the complex. After three, three-hour labs, mutant strains were easily identified by the students and distinguished from wild-type cells analyzing the pattern of SDS-PAGE distribution of proteins. Identifying different subunits of one multimeric protein allowed for discussion of the structure and function of this metabolic enzyme, which captured the interest of the students. The experiment can be adapted to other multimeric protein complexes and shows improvement of the described methodology over previous reports, perhaps because the problem and its solution are representative of the type of techniques currently used in research labs.

Palaeoclimatic conditions in the Late Triassic-Early Jurassic of southern Africa: A geochemical assessment of the Elliot Formation

NASA Astrophysics Data System (ADS)

Sciscio, Lara; Bordy, Emese M.

2016-07-01

The Triassic-Jurassic boundary marks a global faunal turnover event that is generally considered as the third largest of five major biological crises in the Phanerozoic geological record of Earth. Determining the controlling factors of this event and their relative contributions to the biotic turnover associated with it is on-going globally. The Upper Triassic and Lower Jurassic rock record of southern Africa presents a unique opportunity for better constraining how and why the biosphere was affected at this time not only because the succession is richly fossiliferous, but also because it contains important palaeoenvironmental clues. Using mainly sedimentary geochemical proxies (i.e., major, trace and rare earth elements), our study is the first quantitative assessment of the palaeoclimatic conditions during the deposition of the Elliot Formation, a continental red bed succession that straddles the Triassic-Jurassic boundary in southern Africa. Employing clay mineralogy as well as the indices of chemical alteration and compositional variability, our results confirm earlier qualitative sedimentological studies and indicate that the deposition of the Upper Triassic and Lower Jurassic Elliot Formation occurred under increasingly dry environmental conditions that inhibited chemical weathering in this southern part of Pangea. Moreover, the study questions the universal validity of those studies that suggest a sudden increase in humidity for the Lower Jurassic record and supports predictions of long-term global warming after continental flood basalt emplacement.

Forest resources of southern New England

Treesearch

Robert T. Brooks; David B. Kittredge; Carol L. Alerich; Carol L. Alerich

1993-01-01

An analytical report of the third forest inventory of the three southern New England states of Connecticut, Massachusetts, and Rhode Island. Included is a discussion of forest area, number of trees, timber volume, tree biomass, timber value, forest wildlife habitat, ownership, management opportunities, and the future of forest resources in southern New England.

Zonally asymmetric response of the Southern Ocean mixed-layer depth to the Southern Annular Mode

NASA Astrophysics Data System (ADS)

Sallée, J. B.; Speer, K. G.; Rintoul, S. R.

2010-04-01

Interactions between the atmosphere and ocean are mediated by the mixed layer at the ocean surface. The depth of this layer is determined by wind forcing and heating from the atmosphere. Variations in mixed-layer depth affect the rate of exchange between the atmosphere and deeper ocean, the capacity of the ocean to store heat and carbon and the availability of light and nutrients to support the growth of phytoplankton. However, the response of the Southern Ocean mixed layer to changes in the atmosphere is not well known. Here we analyse temperature and salinity data from Argo profiling floats to show that the Southern Annular Mode (SAM), the dominant mode of atmospheric variability in the Southern Hemisphere, leads to large-scale anomalies in mixed-layer depth that are zonally asymmetric. From a simple heat budget of the mixed layer we conclude that meridional winds associated with departures of the SAM from zonal symmetry cause anomalies in heat flux that can, in turn, explain the observed changes of mixed-layer depth and sea surface temperature. Our results suggest that changes in the SAM, including recent and projected trends attributed to human activity, drive variations in Southern Ocean mixed-layer depth, with consequences for air-sea exchange, ocean sequestration of heat and carbon, and biological productivity.

Detection of Rickettsia in Rhipicephalus sanguineus Ticks and Ctenocephalides felis Fleas from Southeastern Tunisia by Reverse Line Blot Assay

PubMed Central

Khrouf, Fatma; M'Ghirbi, Youmna; Znazen, Abir; Ben Jemaa, Mounir; Hammami, Adnene

2014-01-01

Ticks (n = 663) and fleas (n = 470) collected from domestic animals from southeastern Tunisia were screened for Rickettsia infection using reverse line blot assay. Evidence of spotted fever group Rickettsia was obtained. We detected Rickettsia felis in fleas, Rickettsia massiliae Bar 29 and the Rickettsia conorii Israeli spotted fever strain in ticks, and Rickettsia conorii subsp. conorii and Rickettsia spp. in both arthropods. The sensitivity of the adopted technique allowed the identification of a new association between fleas and R. conorii subsp. conorii species. The presence of these vector-borne Rickettsia infections should be considered when diagnosing this disease in humans in Tunisia. PMID:24226919

Predators of the Southern Pine Beetle

Treesearch

John D. Reeve

2011-01-01

This chapter of the Southern Pine Beetle II reviews the overall influence of predators on southern pine beetle (SPB) population dynamics, as well as recent research on specific predators such as the clerid beetle Thanasimus dubius. Several lines of evidence suggest that predators and other natural enemies generate significant SPB mortality that contributes to outbreak...

Southern Sand Dunes

NASA Image and Video Library

2003-01-15

At first glance, this NASA Mars Odyssey image showing impact craters and linear ridges and troughs is typical of the southern highlands. However, upon closer examination migrating sand dunes are observed within the troughs.

Unprecedented Fires in Southern Africa

NASA Technical Reports Server (NTRS)

2002-01-01

The fires that raged across southern Africa this August and September produced a thick 'river of smoke' over the region. NASA-supported studies currently underway on the event will contribute to improved air pollution policies in the region and a better understanding of its impact on climate change. This year the southern African fire season peaked in early September. The region is subject to some of the highest levels of biomass burning in the world. The heaviest burning was in western Zambia, southern Angola, northern Namibia, and northern Botswana. Some of the blazes had fire fronts 20 miles long that lasted for days. In this animation, multiple fires are burning across the southern part of the African continent in September 2000. The fires, indicated in red, were observed by the Advanced Very High Resolution Radiometer (AVHRR) instrument on board the NOAA-14 satellite. The fires generated large amounts of heat-absorbing aerosols (the dark haze), which were observed with the Earth Probe Total Ozone Mapping Spectrometer (TOMS) instrument. These observations were collected as part of a NASA-supported field campaign called SAFARI 2000 (Southern African Regional Science Initiative). The recent six-week 'dry-season' portion of this experiment was planned to coincide with the annual fires. SAFARI 2000 planners tracked the changing location of fires with daily satellite maps provided by researchers at NASA's Goddard Space Flight Center. 'Every year African biomass burning greatly exceeds the scale of the fires seen this year in the western United States,' says Robert Swap of the University of Virginia, one of the campaign organizers. 'But the southern African fire season we just observed may turn out to be an extreme one even by African standards. It was amazing how quickly this region went up in flames.' The thick haze layer from these fires was heavier than campaign participants had seen in previous field studies in the Amazon Basin and during the Kuwati oil fires

Uncaria rhynchophylla Ameliorates Parkinson's Disease by Inhibiting HSP90 Expression: Insights from Quantitative Proteomics.

PubMed

Lan, Yu-Long; Zhou, Jun-Jun; Liu, Jing; Huo, Xiao-Kui; Wang, Ya-Li; Liang, Jia-Hao; Zhao, Jian-Chao; Sun, Cheng-Peng; Yu, Zhen-Long; Fang, Lin-Lin; Tian, Xiang-Ge; Feng, Lei; Ning, Jing; Zhang, Bao-Jing; Wang, Chao; Zhao, Xin-Yu; Ma, Xiao-Chi

2018-06-21

Uncaria rhynchophylla, known as "Gou-teng", is a traditional Chinese medicine (TCM) used to extinguish wind, clear heat, arrest convulsions, and pacify the liver. Although U. rhynchophylla has a long history of being often used to treat central nervous system (CNS) diseases, its efficacy and potential mechanism are still uncertain. This study investigated neuroprotective effect and the underlying mechanism of U. rhynchophylla extract (URE) in MPP+-induced SH-SY5Y cells and MPTP-induced mice. MPP+-induced SH-SY5Y cells and MPTP-induced mice were used to established Parkinson's disease (PD) models. Quantitative proteomics and bioinformatics were used to uncover proteomics changes of URE. Western blotting was used to validate main differentially expressed proteins and test HSP90 client proteins (apoptosis-related, autophagy-related, MAPKs, PI3K, and AKT proteins). Flow cytometry and JC-1 staining assay were further used to confirm the effect of URE on MPP+-induced apoptosis in SH-SY5Y cells. Gait analysis was used to detect the behavioral changes in MPTP-induced mice. The levels of dopamine (DA) and their metabolites were examined in striatum (STR) by HPLC-EC. The positive expression of tyrosine hydroxylase (TH) was detected by immunohischemical staining and Western blotting. URE dose-dependently increased the cell viability in MPP+-induced SH-SY5Y cells. Quantitative proteomics and bioinformatics results confirmed that HSP90 was an important differentially expressed protein of URE. URE inhibited the expression of HSP90, which further reversed MPP+-induced cell apoptosis and autophagy by increasing the expressions of Bcl-2, Cyclin D1, p-ERK, p-PI3K p85, PI3K p110α, p-AKT, and LC3-I and decreasing cleaved caspase 3, Bax, p-JNK, p-p38, and LC3-II. URE also markedly decreased the apoptotic ratio and elevated mitochondrial transmembrane potential (DΨm). Furthermore, URE treatment ameliorated behavioral impairments, increased the contents of DA and its metabolites and

Novel diagnostic procedure for determining metastasis to sentinel lymph nodes in breast cancer using a semi-dry dot-blot method.

PubMed

Otsubo, Ryota; Oikawa, Masahiro; Hirakawa, Hiroshi; Shibata, Kenichiro; Abe, Kuniko; Hayashi, Tomayoshi; Kinoshita, Naoe; Shigematsu, Kazuto; Hatachi, Toshiko; Yano, Hiroshi; Matsumoto, Megumi; Takagi, Katsunori; Tsuchiya, Tomoshi; Tomoshige, Koichi; Nakashima, Masahiro; Taniguchi, Hideki; Omagari, Takeyuki; Itoyanagi, Noriaki; Nagayasu, Takeshi

2014-02-15

We developed an easy, quick and cost-effective detection method for lymph node metastasis called the semi-dry dot-blot (SDB) method, which visualizes the presence of cancer cells with washing of sectioned lymph nodes by anti-pancytokeratin antibody, modifying dot-blot technology. We evaluated the validity and efficacy of the SDB method for the diagnosis of lymph node metastasis in a clinical setting (Trial 1). To evaluate the validity of the SDB method in clinical specimens, 180 dissected lymph nodes from 29 cases, including breast, gastric and colorectal cancer, were examined. Each lymph node was sliced at the maximum diameter and the sensitivity, specificity and accuracy of the SDB method were determined and compared with the final pathology report. Metastasis was detected in 32 lymph nodes (17.8%), and the sensitivity, specificity and accuracy of the SDB method were 100, 98.0 and 98.3%, respectively (Trial 2). To evaluate the efficacy of the SDB method in sentinel lymph node (SLN) biopsy, 174 SLNs from 100 cases of clinically node-negative breast cancer were analyzed. Each SLN was longitudinally sliced at 2-mm intervals and the sensitivity, specificity, accuracy and time required for the SDB method were determined and compared with the intraoperative pathology report. Metastasis was detected in 15 SLNs (8.6%), and the sensitivity, specificity, accuracy and mean required time of the SDB method were 93.3, 96.9, 96.6 and 43.3 min, respectively. The SDB method is a novel and reliable modality for the intraoperative diagnosis of SLN metastasis. © 2013 UICC.

A Secondary Antibody-Detecting Molecular Weight Marker with Mouse and Rabbit IgG Fc Linear Epitopes for Western Blot Analysis.

PubMed

Lin, Wen-Wei; Chen, I-Ju; Cheng, Ta-Chun; Tung, Yi-Ching; Chu, Pei-Yu; Chuang, Chih-Hung; Hsieh, Yuan-Chin; Huang, Chien-Chiao; Wang, Yeng-Tseng; Kao, Chien-Han; Roffler, Steve R; Cheng, Tian-Lu

2016-01-01

Molecular weight markers that can tolerate denaturing conditions and be auto-detected by secondary antibodies offer great efficacy and convenience for Western Blotting. Here, we describe M&R LE protein markers which contain linear epitopes derived from the heavy chain constant regions of mouse and rabbit immunoglobulin G (IgG Fc LE). These markers can be directly recognized and stained by a wide range of anti-mouse and anti-rabbit secondary antibodies. We selected three mouse (M1, M2 and M3) linear IgG1 and three rabbit (R1, R2 and R3) linear IgG heavy chain epitope candidates based on their respective crystal structures. Western blot analysis indicated that M2 and R2 linear epitopes are effectively recognized by anti-mouse and anti-rabbit secondary antibodies, respectively. We fused the M2 and R2 epitopes (M&R LE) and incorporated the polypeptide in a range of 15-120 kDa auto-detecting markers (M&R LE protein marker). The M&R LE protein marker can be auto-detected by anti-mouse and anti-rabbit IgG secondary antibodies in standard immunoblots. Linear regression analysis of the M&R LE protein marker plotted as gel mobility versus the log of the marker molecular weights revealed good linearity with a correlation coefficient R2 value of 0.9965, indicating that the M&R LE protein marker displays high accuracy for determining protein molecular weights. This accurate, regular and auto-detected M&R LE protein marker may provide a simple, efficient and economical tool for protein analysis.

The Southern Ocean biogeochemical divide.

PubMed

Marinov, I; Gnanadesikan, A; Toggweiler, J R; Sarmiento, J L

2006-06-22

Modelling studies have demonstrated that the nutrient and carbon cycles in the Southern Ocean play a central role in setting the air-sea balance of CO(2) and global biological production. Box model studies first pointed out that an increase in nutrient utilization in the high latitudes results in a strong decrease in the atmospheric carbon dioxide partial pressure (pCO2). This early research led to two important ideas: high latitude regions are more important in determining atmospheric pCO2 than low latitudes, despite their much smaller area, and nutrient utilization and atmospheric pCO2 are tightly linked. Subsequent general circulation model simulations show that the Southern Ocean is the most important high latitude region in controlling pre-industrial atmospheric CO(2) because it serves as a lid to a larger volume of the deep ocean. Other studies point out the crucial role of the Southern Ocean in the uptake and storage of anthropogenic carbon dioxide and in controlling global biological production. Here we probe the system to determine whether certain regions of the Southern Ocean are more critical than others for air-sea CO(2) balance and the biological export production, by increasing surface nutrient drawdown in an ocean general circulation model. We demonstrate that atmospheric CO(2) and global biological export production are controlled by different regions of the Southern Ocean. The air-sea balance of carbon dioxide is controlled mainly by the biological pump and circulation in the Antarctic deep-water formation region, whereas global export production is controlled mainly by the biological pump and circulation in the Subantarctic intermediate and mode water formation region. The existence of this biogeochemical divide separating the Antarctic from the Subantarctic suggests that it may be possible for climate change or human intervention to modify one of these without greatly altering the other.

Southern Pine Beetle Behavior and Semiochemistry

Treesearch

Brian T. Sullivan

2011-01-01

The southern pine beetle (SPB) feeds both as adults and larvae within the inner bark of pine trees, which invariably die as a result of colonization. Populations of the SPB erupt periodically and produce catastrophic losses of pines, while at other times the beetles persist almost undetectably in the environment. The southern pine beetle has evolved behaviors that...

Ecological Impacts of Southern Pine Beetle

Treesearch

Maria D. Tchakerian; Robert N. Coulson

2011-01-01

The southern pine beetle (SPB) is the most important biotic disturbance in southern pine forests and causes extensive changes to the forest environment. In this chapter we provide an overview of the ecological impacts of the SPB on forest conditions (the state of the forest) and on forest resources (uses and values associated with the forest). We define ecological...

Metabolic Mapping: Quantitative Enzyme Cytochemistry and Histochemistry to Determine the Activity of Dehydrogenases in Cells and Tissues.

PubMed

Molenaar, Remco J; Khurshed, Mohammed; Hira, Vashendriya V V; Van Noorden, Cornelis J F

2018-05-26

Altered cellular metabolism is a hallmark of many diseases, including cancer, cardiovascular diseases and infection. The metabolic motor units of cells are enzymes and their activity is heavily regulated at many levels, including the transcriptional, mRNA stability, translational, post-translational and functional level. This complex regulation means that conventional quantitative or imaging assays, such as quantitative mRNA experiments, Western Blots and immunohistochemistry, yield incomplete information regarding the ultimate activity of enzymes, their function and/or their subcellular localization. Quantitative enzyme cytochemistry and histochemistry (i.e., metabolic mapping) show in-depth information on in situ enzymatic activity and its kinetics, function and subcellular localization in an almost true-to-nature situation. We describe a protocol to detect the activity of dehydrogenases, which are enzymes that perform redox reactions to reduce cofactors such as NAD(P) + and FAD. Cells and tissue sections are incubated in a medium that is specific for the enzymatic activity of one dehydrogenase. Subsequently, the dehydrogenase that is the subject of investigation performs its enzymatic activity in its subcellular site. In a chemical reaction with the reaction medium, this ultimately generates blue-colored formazan at the site of the dehydrogenase's activity. The formazan's absorbance is therefore a direct measure of the dehydrogenase's activity and can be quantified using monochromatic light microscopy and image analysis. The quantitative aspect of this protocol enables researchers to draw statistical conclusions from these assays. Besides observational studies, this technique can be used for inhibition studies of specific enzymes. In this context, studies benefit from the true-to-nature advantages of metabolic mapping, giving in situ results that may be physiologically more relevant than in vitro enzyme inhibition studies. In all, metabolic mapping is an

The Southern California Twin Register at the University of Southern California: III

PubMed Central

Baker, Laura A.; Tuvblad, Catherine; Wang, Pan; Gomez, Karina; Bezdjian, Serena; Niv, Sharon; Raine, Adrian

2013-01-01

The Southern California Twin Register at the University of Southern California (USC) was initiated in 1984 and continues to provide an important resource for studies investigating genetic and environmental influences on human behavior. This article provides an update on the current register and its potential for future twin studies using recruitment through school district databases and voter records. An overview is also provided for an ongoing longitudinal twin study investigating the development of externalizing psychopathology from childhood to young adulthood, the USC Study of Risk Factors for Antisocial Behavior. Characteristics of the twins and their families are presented, including recruitment and participation rates, as well as attrition analyses and a summary of key findings to date. PMID:23394193

The influence of annual precipitation, topography, and vegetative cover on soil moisture and summer drought in southern California.

PubMed

Miller, P C; Poole, D K

1983-02-01

The influence of annual precipitation and vegetation cover on soil moisture and on the length of the summer drought was estimated quantitatively using 9 years of soil moisture data collected at Echo Valley in southern California. The measurements support the conclusions that in the semi-arid mediterranean climate a soil drought will occur regardless of vegetation cover and annual precipitation, but the length of the drought is greatly dependent on soil depth and rockiness. Evergreen species which can survive this drought tend to accentuate the drought, especially in deep soil levels, by developing a canopy with a large transpiring surface.

Smoke in Southern Africa

NASA Technical Reports Server (NTRS)

2002-01-01

This SeaWiFS true-color image acquired over Southern Africa on Sept. 4, 2000, shows a thick shroud of smoke and haze blanketing much of the southern half of the continent. The smoke in this scene is being generated by a tremendous number of fires burning over a large area across the countries of Angola, Zambia, Mozambique, Zimbabwe, Botswana, and the Northern Province of South Africa. In this image, the smoke (grey pixels) is easily distinguished from clouds (bright white pixels). Refer to the Images and Data section for a larger scale view of the fires in Southern Africa. Data from both the SeaWiFS and Terra satellites are being used by an international team of scientists participating in the SAFARI field experiment. The objective of SAFARI is to measure the effects of windblown smoke and dust on air quality and the Earth's radiant energy budget. This image was produced using SeaWiFS channels 6, 5, and 1 (centered at 670 nm, 555 nm , and 412 nm, respectively). The data were acquired and provided by the Satellite Applications Center in Pretoria, South Africa. Image courtesy Gene Feldman, SeaWiFS Project and Orbital Sciences

Northerners versus southerners: Italian anthropology and psychology faced with the "southern question".

PubMed

Cimino, Guido; Foschi, Renato

2014-11-01

Following the Unification of Italy (1861), when confronted with the underdevelopment problems of the south that had given rise to the so-called "southern question," some Italian anthropologists and psychologists began to study the populations of the south from the psycho-anthropological point of view. These scientists, at times subject to preconceived ideas toward the southerners, conveyed observations and descriptions of the southern character traits that, in general, were considered different, in a negative sense, with respect to those of the northern peoples. To explain such diversity in the "psychological" characteristics between the north and south of the country (presumed cause also of the south's backwardness), various hypotheses were advanced related to the kind of heredity theory adopted, which could be of, more or less, an "innatist" or "transformist" or "environmentalist" kind. The distinction proposed in this article between at least 2 different "hereditarian" theories formulated by the Italian scientists, and the confrontation of these theories with the hypotheses expressed by the "southernist" sociologists, contrary to the idea of "racial varieties" present in the Italian population, allows one to understand in what way and in what sense, at the threshold of the 20th century, there arose the ideology of "Nordicism" and the roots of racism were planted.

Geomorphical and Geochronological Constrains of the Last Glacial Period in Southern Patagonia, Southern South America

NASA Astrophysics Data System (ADS)

García, J.; Hall, B. L.; Kaplan, M. R.; Vega, R. M.; Binnie, S. A.; Hein, A.; Gómez, G. N.; Ferrada, J. J.

2013-12-01

Despite the outer limits of the former Patagonian ice sheet (PIS, ~38-55S) having been extensively mapped, it remains unknown if the Patagonian glaciers fluctuated synchronously or asynchronously during the last glacial period. Previous work has revealed asynchronous spatiotemporal ice dynamics along the eastern and western ice-margins at the end of the last glaciation but it is not well understood if the northern and southern parts of the PIS reached concurrent maximum glaciation during the last glacial cycle. The Patagonian Andes is the only landmass involving the southern westerly wind belt latitudinal range, which is thought to have played a key role in past glacial and climate changes. Therefore, reconstructing southern Andes glacier history constitutes a key element for understanding the cause of glaciations in Patagonia and the role of the westerlies in climate change. Here, we discuss paleoglaciological and paleoclimatological implications of new 10Be and 14C data obtained from moraines and strategically selected mires in two contiguous glacially molded basins of south Patagonia (48-55S): Torres del Paine (51S) and Última Esperanza (52S). In this region, we focused our 10Be cosmogenic-dating efforts in the previously undated outer moraines deposited (supposedly) during the last glacial cycle. In order to crosscheck cosmogenic data we collected boulders embedded in moraines and cobbles from the main glaciofluvial plains grading from the outermost moraines. Geomorphic and cosmogenic dating affords evidence for glacial maximum conditions occurring between 40-50 ka (ka = thousand of years before present) in southern Patagonia, which is different from other chronologies within southern South America. We obtained 14C basal ages from sites located within moraine depressions and on former paleolake shorelines and thus these may provide key data on deglaciation and debated regional paleolake history.

Eddies in the Southern Ocean

NASA Image and Video Library

2015-04-08

The cloud cover over the Southern Ocean occasionally parts as it did on January 1, 2015 just west of the Drake Passage where the VIIRS instrument on the Suomi NPP satellite glimpsed the above collection of ocean-color delineated eddies which have diameters ranging from a couple of kilometers to a couple of hundred kilometers. Recent studies indicate that eddy activity has been increasing in the Southern Ocean with possible implications for climate change. Credit: NASA's OceanColor/Suomi NPP/VIIRS

Developing a chironomid training set for western South America (South-Central Chile): potential for quantitative temperature reconstructions

NASA Astrophysics Data System (ADS)

Araneda, A.; Larocque-Tobler, I.; Torrejon, F.; Grosjean, M.; Jana-Pinninghoff, P.; Ortega, C.; Urrutia, R.

2012-12-01

Quantitative climate reconstructions of the last two millennia are a fundamental issue in order to compare the current trends in climate observed nowadays. At global scale most of the climate reconstructions have been developed for the Northern Hemisphere, while for the Southern Hemisphere quantitative reconstructions are very rare and very limited geographically. The recognition of such disparity has generated among other research initiatives the LOTRED-SA Long-Term climate Reconstruction and Dynamics of (southern) South America, a collaborative, high-resolution multi-proxy approach within the framework of the IGBP-PAGES program. In this context our work presents the results of a 50-lakes training set in Central-Southern Chile developed with the aim to generate a basis for quantitative chironomid-inferred temperature reconstructions for this part of the continent. Chironomids (Insecta: Diptera) are aquatic insects that develop a great proportion of their life cycle as larvae in aquatic ecosystems. Several studies, developed mainly in the Northern Hemisphere, have proven their usefulness in reconstructing past climate due to the larvae's relationship to temperature. The training set developed here includes lakes located between 34 and 48 S, covering a broad temperature (as latitudinal) gradient. The surface (0-1 cm) sediment of each lake was sampled and chironomids, organic matter and nutrient were analyzed. Water analyses included the measurement of 10 variables (AirT, WBT, WST, N-tot, P-tot, Fe, Na, pH among others). In order to identify the most important variables explaining the highest variance in the chironomid assemblages, ordinations analyses were performed. A preliminary DCA analysis indicated, according to the length of gradients smaller than 3 STD, that a linear model was more appropriate for further analysis. Hence a RDA analysis was applied to the environmental and species data, indicating that the most important variables to determine chironomid

A methodology for assessing annual risk of southern pine beetle outbreaks across the southern region using pheromone traps

Treesearch

Ronald F Billings; William W. Upton

2010-01-01

An operational system to forecast infestation trends (increasing, static, declining) and relative population levels (high, moderate, low) of the southern pine beetle (SPB), Dendroctonus frontalis, has been implemented in the Southern and Eastern United States. Numbers of dispersing SPB and those of a major predator (the clerid beetle, ...

Quaternary paleoecology of aquatic Diptera in tropical and Southern Hemisphere regions, with special reference to the Chironomidae

NASA Astrophysics Data System (ADS)

Verschuren, Dirk; Eggermont, Hilde

2006-08-01

Chironomid paleoecology in north-temperate regions has made tremendous progress over the past decade, but studies in tropical and Southern Hemisphere regions remain relatively scarce. Reasons for this imbalance are (1) incomplete taxonomic knowledge of chironomid faunas outside Europe and North America, (2) a scarcity of ecological data on local species and genera that might confer bio-indicator value to them, and (3) logistic difficulties hampering the lake surveying necessary to develop paleoenvironmental calibration data sets. Thus far, most chironomid paleoecology in tropical and Southern Hemisphere regions has relied on an indicator-species approach, combining autecological information on local species of which larval morphology is known with the traditional ecological typology of chironomid genera transferred from the Holarctic region. This paper reviews work accomplished to date in tropical and temperate South America, Australia, Africa, and New Zealand, including studies on various families of non-chironomid Diptera with diagnostic fossils. Research has focused mostly on late-Glacial and Holocene climate reconstruction, less on tracing past human disturbance of aquatic ecosystems and their drainage basins. Quantitative chironomid-based paleoenvironmental reconstruction has so far been done only in Australia and Africa. These studies compensated for the lack of traditional surface-sediment calibration data sets, nowadays often the main source of quantitative information on species ecological optima and tolerances, by maximally exploiting archival species-distribution data based on live collections of adult and/or larval midges. This stimulated efforts to achieve trustworthy species-level identification of fossil chironomid remains, and, as a result, the taxonomic resolution of paleoecological studies in Australia and Africa is higher on average than that achieved in European and North American studies.

Progressive approach to eruption at Campi Flegrei caldera in southern Italy

NASA Astrophysics Data System (ADS)

Kilburn, Christopher R. J.; de Natale, Giuseppe; Carlino, Stefano

2017-05-01

Unrest at large calderas rarely ends in eruption, encouraging vulnerable communities to perceive emergency warnings of volcanic activity as false alarms. A classic example is the Campi Flegrei caldera in southern Italy, where three episodes of major uplift since 1950 have raised its central district by about 3 m without an eruption. Individual episodes have conventionally been treated as independent events, so that only data from an ongoing episode are considered pertinent to evaluating eruptive potential. An implicit assumption is that the crust relaxes accumulated stress after each episode. Here we apply a new model of elastic-brittle failure to test the alternative view that successive episodes promote a long-term accumulation of stress in the crust. The results provide the first quantitative evidence that Campi Flegrei is evolving towards conditions more favourable to eruption and identify field tests for predictions on how the caldera will behave during future unrest.

Progressive approach to eruption at Campi Flegrei caldera in southern Italy.

PubMed

Kilburn, Christopher R J; De Natale, Giuseppe; Carlino, Stefano

2017-05-15

Unrest at large calderas rarely ends in eruption, encouraging vulnerable communities to perceive emergency warnings of volcanic activity as false alarms. A classic example is the Campi Flegrei caldera in southern Italy, where three episodes of major uplift since 1950 have raised its central district by about 3 m without an eruption. Individual episodes have conventionally been treated as independent events, so that only data from an ongoing episode are considered pertinent to evaluating eruptive potential. An implicit assumption is that the crust relaxes accumulated stress after each episode. Here we apply a new model of elastic-brittle failure to test the alternative view that successive episodes promote a long-term accumulation of stress in the crust. The results provide the first quantitative evidence that Campi Flegrei is evolving towards conditions more favourable to eruption and identify field tests for predictions on how the caldera will behave during future unrest.

The Southern Forest Futures Project: technical report

Treesearch

David N. Wear; John G. Greis

2013-01-01

Please visit the Southern Forest Futures Project website for more information.The Southern Forest Futures Project provides a science-based “futuring” analysis of the forests of the 13 States of the Southeastern United States. With findings...

Steam-bending properties of southern pine

Treesearch

Truett J. Lemoine; Peter Koch

1971-01-01

Southern pine wood can be successfully steam-bent if the bending jig incorporates a flexible metal bending strap together with a mechanism to apply a uniform end compression load during the bending operation. With clear, 1/2- and 1-inch-thick southern pine at 17-percent moisture content, highest bending rating where obtained with fast-grown, vertical-grain, low-density...

Pteropods in Southern Ocean ecosystems

NASA Astrophysics Data System (ADS)

Hunt, B. P. V.; Pakhomov, E. A.; Hosie, G. W.; Siegel, V.; Ward, P.; Bernard, K.

2008-09-01

To date, little research has been carried out on pelagic gastropod molluscs (pteropods) in Southern Ocean ecosystems. However, recent predictions are that, due to acidification resulting from a business as usual approach to CO 2 emissions (IS92a), Southern Ocean surface waters may begin to become uninhabitable for aragonite shelled thecosome pteropods by 2050. To gain insight into the potential impact that this would have on Southern Ocean ecosystems, we have here synthesized available data on pteropod distributions and densities, assessed current knowledge of pteropod ecology, and highlighted knowledge gaps and directions for future research on this zooplankton group. Six species of pteropod are typical of the Southern Ocean south of the Sub-Tropical Convergence, including the four Thecosomes Limacina helicina antarctica, Limacina retroversa australis, Clio pyramidata, and Clio piatkowskii, and two Gymnosomes Clione limacina antarctica and Spongiobranchaea australis. Limacina retroversa australis dominated pteropod densities north of the Polar Front (PF), averaging 60 ind m -3 (max = 800 ind m -3) and 11% of total zooplankton at the Prince Edward Islands. South of the PF L. helicina antarctica predominated, averaging 165 ind m -3 (max = 2681 ind m -3) and up to >35% of total zooplankton at South Georgia, and up to 1397 ind m -3 and 63% of total zooplankton in the Ross Sea. Combined pteropods contributed <5% to total zooplankton in the Lazarev Sea, but 15% (max = 93%) to macrozooplankton in the East Antarctic. In addition to regional density distributions we have synthesized data on vertical distributions, seasonal cycles, and inter-annual density variation. Trophically, gymnosome are specialist predators on thecosomes, while thecosomes are considered predominantly herbivorous, capturing food with a mucous web. The ingestion rates of L. retroversa australis are in the upper range for sub-Antarctic mesozooplankton (31.2-4196.9 ng pig ind -1 d -1), while those of L

Surface uplift and atmospheric flow deflection in the Late Cenozoic southern Sierra Nevada

NASA Astrophysics Data System (ADS)

Mix, H.; Caves, J. K.; Winnick, M.; Ritch, A. J.; Reilly, S.; Chamberlain, C. P.

2016-12-01

Given the intimate links between topography, tectonics, climate and biodiversity, considerable effort has been devoted to developing robust elevation histories of orogens. In particular, quantitative geochemical reconstructions using stable oxygen and hydrogen isotopes have been applied to many of the world's mountain belts. Yet after decades of study, determining the Cenozoic surface uplift history of the Sierra Nevada remains a challenge. While geological and geophysical evidence suggests the southern Sierra underwent 1-2 km of Late Cenozoic surface uplift, stable isotope paleoaltimetry studies to date have been restricted to the Basin and Range interior. Recent advances in atmospheric modeling have suggested that such stable isotope records from leeward sites can be affected by the complicating role that sufficiently elevated topography such as the southern (High) Sierra plays in diverting atmospheric circulation. In order to examine the potential role of these terrain blocking effects, we produced stable isotope records from three Late Cenozoic sedimentary basins in the Eastern Sierra and Basin and Range: 1) Authigenic clay minerals in the Mio-Pliocene Verdi Basin (VB), 2) Fluvial and lacustrine carbonates from the Plio-Pleistocene Coso Basin (CB), and 3) Miocene to Holocene pedogenic, fluvial and lacustrine carbonates of Fish Lake Valley (FLV). Whereas both the VB (near present-day Reno) and CB (southern Owens Valley) receive input of water directly from the Sierra crest, FLV is a region of proposed reconvergence of moisture in the Basin and Range. The oxygen isotope records in both CB and FLV increase during the Neogene by approximately 2 ‰, while the hydrogen isotope record of the VB decreases by <10 ‰. These results are consistent with a modestly-elevated Paleogene Sierra of 2 km over which air masses traversed and underwent orographic rainout and Rayleigh distillation. A Neogene pulse of uplift in the southern Sierra could have driven modern flow

Excitation of the Earth's Chandler wobble by southern oscillation/El Nino, 1900-1979

NASA Technical Reports Server (NTRS)

Chao, B. F.

1985-01-01

The southern oscillation/El Nino (ENSO) is the single most prominent interannual signal in global atmospheric/oceanic fluctuations. The following question is addressed: how important is the angular momentum carried by ENSO in exciting the Earth's Chandler wobble? The question is attacked through a statistical analysis of the coherence spectra (correlation as a function of frequency) between two data sets spanning 1900 to 1979-the southern oscillation index (SOI) time series and the excitation function psi (with x-component psi sub x and y-component psi sub y) of the Chandler wobble derived from the homogeneous ILS (International Latitude Service) polar motion data. The coherence power and phase in the Chandler frequency band (approx. 0.79 to 0.89 cpy) are studied. It is found that, during 1900 to 1979 the coherence between SOI and psi sub x is significant well over the 95% confidence threshold whereas that between SOI and psi sub y is practically nil. Quantitatively, the coherence study shows that ENSO provides some 20% of the observed Chandler wobble excitation power. Since earlier investigations have shown that the total atmospheric/oceanic variation can account for the Chandler wobble excitation at about 20% level, the implication is that ENSO maybe an important (interannual) part of the atmospheric/oceanic variation that is responsible for the Chandler wobble excitation during 1900 to 1979.

Quantitative nephelometry

MedlinePlus

... this page: //medlineplus.gov/ency/article/003545.htm Quantitative nephelometry test To use the sharing features on this page, please enable JavaScript. Quantitative nephelometry is a lab test to quickly and ...

Ethics Instruction in Community College Leadership Programs: Southern Perspectives

ERIC Educational Resources Information Center

Ware, Nikisha Green

2011-01-01

The purpose of this study was to discover which southern universities have graduate preparatory programs in community college leadership and how, if at all, ethics is addressed in their curricula and in instruction. Surveys were mailed to 38 southern universities located in the Southern Regional Education Board member states. Of the 21 responses…

Sexty Southerners: Sexting Content and Behaviors among Selected Southern Undergraduates

ERIC Educational Resources Information Center

Hudson, Heather K.; Marshall, S. Alexandra

2016-01-01

Sexting is defined as sending/posting/sharing sexually explicit messages or nude/semi-nude images via electronic communication. Purpose: The purpose of this study was to assess and determine relationships of sexting behavioral intentions, sexting behaviors, and sexting content among selected Southern undergraduate students. Methods: Survey…

Stem Profile for Southern Equations for Southern Tree Species

Treesearch

Alexander Clark; Ray A. Souter; Bryce E. Schlaegel

1991-01-01

Form-class segmented-profile equations for 58 southern tree species and species groups are presented.The profile equations are based on taper data for 13,469 trees sampled in natural stands in many locations across the South.The profile equations predict diameter at any given height, height to give diameter, and volume between two heights.Equation coefficients for use...

Recombinant expression of rt-PA gene (encoding Reteplase) in gametophytes of the seaweed Laminaria japonica (Laminariales, Phaeophyta).

PubMed

Zhang, YiChen; Jiang, Peng; Gao, JiangTao; Liao, JianMin; Sun, ShiJing; Shen, ZiLong; Qin, Song

2008-12-01

The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte. The expression of foreign genes in sporophte has been proved. In this research, the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator (Reteplase), which is a thrombolytic agent for acute myocardial infarction (AMI). Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin (PPT), and proliferated in an established bubble column photo-bioreactor. According to the results from quantitative ELISA, Southern blotting, and fibrin agarose plate assay (FAPA) for bioactivity, it was showed that the rt-PA gene had been integrated into the genome of gametophytes of L. japonica, and the expression product showed the expected bioactivity, implying the proper post-transcript modification in haploid gametophyte.

Stand density management of southern bottomland hardwoods

Treesearch

J.C.G. Goelz; J.S. Meadows

1997-01-01

We present tools to guide stand density management of southern bottomiand hardwoods and we provide guidance in their implementation. We present stocking guides for southern bottomland hardwoods and variants for associated forest types, water tupelo (Nyssa aquarica L.) and sweetgum (Liquidumbar styraciflua L.). The A-line...

50 CFR 223.211 - Southern DPS of spotted seal.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 50 Wildlife and Fisheries 9 2011-10-01 2011-10-01 false Southern DPS of spotted seal. 223.211... Applicable to Threatened Marine and Anadromous Species § 223.211 Southern DPS of spotted seal. The... species shall apply to the Southern Distinct Population Segment of the spotted seal listed in § 223.102(a...

Silviculture and management strategies applicable to southern upland hardwoods

Treesearch

Ray R. Hicks; Deborah K. Kennard; H. Michael Rauscher; Daniel L. Schmoldt; Patricia A. Flebbe

2001-01-01

The southern upland hardwoods include extensive areas in the southern Appalachians, Cumberland Plateau and Ozark/Ouachita regions. The majority of commercial hardwoods in the south occur in the region often referred to as the "Southern Appalachian Region". For purposes of this discussion, this region includes the hilly or mountainous area west and north of...

Clinical usefulness of Western blotting and ELISA avidity for the diagnosis of human toxocariasis.

PubMed

Rudzińska, M; Kowalewska, B; Sikorska, K

2017-01-01

The serodiagnosis of human toxocariasis is difficult. Specific IgGs detected routinely with ELISA based on Toxocara excretory-secretory (TES) antigens often persist for years at an elevated level, which does not allow either the differentiation between an active and persistent infection or monitoring of the effect of treatment. Additionally, false-positive results may occur in co-infections with other helminths due to cross-reactions. We evaluated the usefulness of an IgG avidity index (AI) and a Western blotting (WB) IgG in the diagnosis of patients suspected of Toxocara infection. We studied 138 subjects who were submitted to serological testing two or more times. Confirmation of an infection by WB was achieved in 73.2% of patients. A high AI was obtained in 89.1% of patients, and low AI and borderline AI were found in only 10.9%. Low and borderline values of AI remained at similar levels in subsequent studies over 2-3 years. The results showed the necessity of obligatory verification of all ELISA IgG positive and questionable results by WB. The index of IgG avidity may be helpful in excluding recent infection, but its usefulness in detecting an active phase of invasion requires further research. © 2016 John Wiley & Sons Ltd.

Simulating the impacts of southern pine beetle and fire on the dynamics of xerophytic pine landscapes in the southern Appalachians

Treesearch

J.D. Waldron; C.W. Lafon; R.N. Coulson; D.M. Cairns; M.D. Tchakerian; A. Birt; K.D. Klepzig

2007-01-01

Question: Can fire be used to maintain Yellow pine (Pinus subgenus Diploxylon) stands disturbed by periodic outbreaks of southern pine beetle?Location: Southern Appalachian Mountains, USA.Methods: We used LANDIS to model vegetation disturbance and succession...

Comparison of the performance in detection of HPV infections between the high-risk HPV genotyping real time PCR and the PCR-reverse dot blot assays.

PubMed

Zhang, Lahong; Dai, Yibei; Chen, Jiahuan; Hong, Liquan; Liu, Yuhua; Ke, Qiang; Chen, Yiwen; Cai, Chengsong; Liu, Xia; Chen, Zhaojun

2018-01-01

A new multiplex real-time PCR assay, the high-risk HPV genotyping real time PCR assay (HR HPV RT-PCR), has been developed to detect 15 high-risk HPV types with respective viral loads. In this report, a total of 684 cervical specimens from women diagnosed with vaginitis were assessed by the HR HPV RT-PCR and the PCR reaction and reverse dot blot (PCR-RDB) assays, using a PCR-sequencing method as a reference standard. A total coincidence of 97.7% between the HR HPV RT PCR and the PCR-RDB assays was determined with a Kappa value of 0.953. The HR HPV RT PCR assay had sensitivity, specificity, and concordance rates (accuracy) of 99.7%, 99.7%, and 99.7%, respectively, as confirmed by PCR-sequencing, while the PCR-RDB assay had respective rates of 98.8%, 97.1%, and 98.0%. The overall rate of HPV infection, determined by PCR-sequencing, in women diagnosed with vaginitis was 49.85%, including 36.26% of single infection and 13.6% of multiple infections. The most common infections among the 15 high-risk HPV types in women diagnosed with vaginitis were HPV-52, HPV-16, and HPV-58, with a total detection rate of 10.23%, 7.75%, and 5.85%, respectively. We conclude that the HR HPV RT PCR assay exhibits better clinical performance than the PCR-RDB assay, and is an ideal alternative method for HPV genotyping. In addition, the HR HPV RT PCR assay provides HPV DNA viral loads, and could serve as a quantitative marker in the diagnosis and treatment of single and multiple HPV infections. © 2017 Wiley Periodicals, Inc.

Southern pine beetle infestations in relation to forest stand conditions, previous thinning, and prescribed burning: evaluation of the Southern Pine Beetle Prevention Program

Treesearch

John T. Nowak; James R. Meeker; David R. Coyle; Chris A. Steiner; Cavell Brownie

2015-01-01

Since 2003, the Southern Pine Beetle Prevention Program (SPBPP) (a joint effort of the USDA Forest Service and Southern Group of State Foresters) has encouraged and provided cost-share assistance for silvicultural treatments to reduce stand/forest susceptibility to the southern pine beetle (SPB)(Dendroctonus frontalis Zimmermann) in the southeastern United States....

Human Hrs, a tyrosine kinase substrate in growth factor-stimulated cells: cDNA cloning and mapping of the gene to chromosome 17.

PubMed

Lu, L; Komada, M; Kitamura, N

1998-06-15

Hrs is a 115kDa zinc finger protein which is rapidly tyrosine phosphorylated in cells stimulated with various growth factors. We previously purified the protein from a mouse cell line and cloned its cDNA. In the present study, we cloned a human Hrs cDNA from a human placenta cDNA library by cross-hybridization, using the mouse cDNA as a probe, and determined its nucleotide sequence. The human Hrs cDNA encoded a 777-amino-acid protein whose sequence was 93% identical to that of mouse Hrs. Northern blot analysis showed that the Hrs mRNA was about 3.0kb long and was expressed in all the human adult and fetal tissues tested. In addition, we showed by genomic Southern blot analysis that the human Hrs gene was a single-copy gene with a size of about 20kb. Furthermore, the human Hrs gene was mapped to chromosome 17 by Southern blotting of genomic DNAs from human/rodent somatic cell hybrids. Copyright 1998 Elsevier Science B.V. All rights reserved.

Compositions and methods for detecting gene rearrangements and translocations

DOEpatents

Rowley, Janet D.; Diaz, Manuel O.

2000-01-01

Disclosed is a series of nucleic acid probes for use in diagnosing and monitoring certain types of leukemia using, e.g., Southern and Northern blot analyses and fluorescence in situ hybridization (FISH). These probes detect rearrangements, such as translocations involving chromosome band 11q23 with other chromosomes bands, including 4q21, 6q27, 9p22, 19p13.3, in both dividing leukemic cells and interphase nuclei. The breakpoints in all such translocations are clustered within an 8.3 kb BamHI genomic region of the MLL gene. A novel 0.7 kb BamH1 cDNA fragment derived from this gene detects rearrangements on Southern blot analysis with a single BamHI restriction digest in all patients with the common 11q23 translocations and in patients with other 11q23 anomalies. Northern blot analyses are presented demonstrating that the MLL gene has multiple transcripts and that transcript size differentiates leukemic cells from normal cells. Also disclosed are MLL fusion proteins, MLL protein domains and anti-MLL antibodies.

Diagnostic potential of Western blot analysis of sera from dogs with leishmaniasis in endemic areas and significance of the pattern.

PubMed

Aisa, M J; Castillejo, S; Gallego, M; Fisa, R; Riera, M C; de Colmenares, M; Torras, S; Roura, X; Sentis, J; Portus, M

1998-02-01

Serum samples collected from 237 dogs in Catalonia (northeastern Spain) were screened by Western blot analysis to detect the presence of antibodies specific to different Leishmania infantum polypeptide fractions. Leishmaniasis was confirmed in 72 of these dogs by direct examination and/or culture. Another 165 animals from the Priorat region were studied periodically for 2-8 years between 1987 and 1995, giving a total of 565 determinations. A control group of 93 dogs from nonendemic areas was also studied. Sera from dogs with leishmaniasis recognized antigens with molecular weights ranging from 12 to 85 kD. The most sensitive antigens were those of 70, 65, 46, 30, 28, 14, and 12 kD, which were recognized by 75%, 75%, 78%, 75%, 81%, 79%, and 75%, respectively, of the sera from dogs with positive parasitologic examination results. Antigens of 70 and 65 kD were also recognized by two dogs from nonendemic areas. Antigens of 14 and 12 kD were the first to be recognized by sera of asymptomatic dogs with titers less than the cut-off value of the dot-ELISA that increased during the longitudinal study, and the presence of antibodies specific for these fractions was observed for up to six years before seroconversion observed by dot-ELISA. These antibodies were also the first to disappear in dogs in which the disease was self-limited. The study corroborates the high sensitivity and specificity of Western blots in the diagnosis of canine leishmaniasis when the bands of low molecular weight (less than 46 kD) are considered, and indicates that fractions of 14 and 12 kD are useful in detecting early forms of the disease.

A Secondary Antibody-Detecting Molecular Weight Marker with Mouse and Rabbit IgG Fc Linear Epitopes for Western Blot Analysis

PubMed Central

Cheng, Ta-Chun; Tung, Yi-Ching; Chu, Pei-Yu; Chuang, Chih-Hung; Hsieh, Yuan-Chin; Huang, Chien-Chiao; Wang, Yeng-Tseng; Kao, Chien-Han; Roffler, Steve R.; Cheng, Tian-Lu

2016-01-01

Molecular weight markers that can tolerate denaturing conditions and be auto-detected by secondary antibodies offer great efficacy and convenience for Western Blotting. Here, we describe M&R LE protein markers which contain linear epitopes derived from the heavy chain constant regions of mouse and rabbit immunoglobulin G (IgG Fc LE). These markers can be directly recognized and stained by a wide range of anti-mouse and anti-rabbit secondary antibodies. We selected three mouse (M1, M2 and M3) linear IgG1 and three rabbit (R1, R2 and R3) linear IgG heavy chain epitope candidates based on their respective crystal structures. Western blot analysis indicated that M2 and R2 linear epitopes are effectively recognized by anti-mouse and anti-rabbit secondary antibodies, respectively. We fused the M2 and R2 epitopes (M&R LE) and incorporated the polypeptide in a range of 15–120 kDa auto-detecting markers (M&R LE protein marker). The M&R LE protein marker can be auto-detected by anti-mouse and anti-rabbit IgG secondary antibodies in standard immunoblots. Linear regression analysis of the M&R LE protein marker plotted as gel mobility versus the log of the marker molecular weights revealed good linearity with a correlation coefficient R2 value of 0.9965, indicating that the M&R LE protein marker displays high accuracy for determining protein molecular weights. This accurate, regular and auto-detected M&R LE protein marker may provide a simple, efficient and economical tool for protein analysis. PMID:27494183

Blue-stain Fungi Associated with Roots of Southern Pine Trees Attacked by the Southern Pine Beetle, Dendroctonus frontalis

Treesearch

William J. Otrosina; Nolan J. Hess; Stanley J. Zarnoch; Thelma J. Perry; John P. Jones

1997-01-01

Forty paired plots were established from eastern Texas to Alabama to study root-infecting, blue-stain fungi in southern pine stands undergoing southern pine beetle (SPB) attack. Woody roots were sampled in plots undergoing recent or current attack by the SPB. Comparisons were made between occurrence of Lcptogrqhiumspp. and related fungi and data on various...

2014 Southern Nevada Household Travel Survey | Transportation Secure Data

Science.gov Websites

Center | NREL 14 Southern Nevada Household Travel Survey 2014 Southern Nevada Household Travel Survey The 2014 Southern Nevada Household Travel Survey collected information from residents in the Las conduct the survey. Methodology The survey was conducted in two phases-from March to May 2014 and from

The effect of exchange rates on southern pine exports

Treesearch

H.W. Wisdom; James E. Granskog

2003-01-01

Changes in exchange rates affect southern pine exports by changing the cost of southern wood in foreign markets. A strong dollar discourages exports; a weak dollar encourages exports. A simple economic export market model is developed to determine whether changes in the exchange rates in foreign markets of southern pine products have, in fact, let to significant...

Southern hardwood forestry group going strong after 50 years

Treesearch

Brian Roy Lockhart; Steve Meadows; Jeff Portwood

2005-01-01

On November 15,200 1, the Southern Hardwood Forestry Group (referred to as the Group) met at the U.S. Forest Service Southern Research Station's Southern Hardwoods Laboratory in Stoneville, hlississippi to celebrate the Group's 50th anniversary. About 130 members and guests attended to celebrate the 50th anniversary of the Group and to honor its charter...

Southern Appalachian Man and the Biosphere - SAMAB.ORG

Science.gov Websites

Southern Appalachian Man and the Biosphere Home Who We Are SAMAB Initiatives Archives Welcome to SAMAB This slideshow requires JavaScript. The Southern Appalachian Man and the Biosphere (SAMAB

Are neoclassical canons valid for southern Chinese faces?

PubMed

Jayaratne, Yasas S N; Deutsch, Curtis K; McGrath, Colman P J; Zwahlen, Roger A

2012-01-01

Proportions derived from neoclassical canons, initially described by Renaissance sculptors and painters, are still being employed as aesthetic guidelines during the clinical assessment of the facial morphology. 1. to determine the applicability of neoclassical canons for Southern Chinese faces and 2. to explore gender differences in relation to the applicability of the neoclassical canons and their variants. 3-D photographs acquired from 103 young adults (51 males and 52 females) without facial dysmorphology were used to test applicability of four neoclassical canons. Standard anthropometric measurements that determine the facial canons were made on these 3-D images. The validity of the canons as well as their different variants were quantified. The neoclassical cannons seldom applied to these individuals, and facial three-section and orbital canons did not apply at all. The orbitonasal canon was most frequently applicable, with a frequency of 19%. Significant sexual dimorphism was found relative to the prevalence of the variants of facial three-section and orbitonasal canons. The neoclassical canons did not appear to apply to our sample when rigorous quantitative measurements were employed. Thus, they should not be used as esthetic goals for craniofacial surgical interventions.

Are Neoclassical Canons Valid for Southern Chinese Faces?

PubMed Central

Jayaratne, Yasas S. N.; Deutsch, Curtis K.; McGrath, Colman P. J.; Zwahlen, Roger A.

2012-01-01

Background Proportions derived from neoclassical canons, initially described by Renaissance sculptors and painters, are still being employed as aesthetic guidelines during the clinical assessment of the facial morphology. Objective 1. to determine the applicability of neoclassical canons for Southern Chinese faces and 2. to explore gender differences in relation to the applicability of the neoclassical canons and their variants. Methodology 3-D photographs acquired from 103 young adults (51 males and 52 females) without facial dysmorphology were used to test applicability of four neoclassical canons. Standard anthropometric measurements that determine the facial canons were made on these 3-D images. The validity of the canons as well as their different variants were quantified. Principal Findings The neoclassical cannons seldom applied to these individuals, and facial three-section and orbital canons did not apply at all. The orbitonasal canon was most frequently applicable, with a frequency of 19%. Significant sexual dimorphism was found relative to the prevalence of the variants of facial three-section and orbitonasal canons. Conclusion The neoclassical canons did not appear to apply to our sample when rigorous quantitative measurements were employed. Thus, they should not be used as esthetic goals for craniofacial surgical interventions. PMID:23285105

Channeled Southern Highlands

NASA Image and Video Library

2018-04-30

This enhanced color image from NASA's Mars Reconnaissance Orbiter (MRO) shows the heavily channeled and ancient southern highlands of Mars. The elongated and jagged features are windblown dunes, perhaps hardened and eroded. For more information see https://photojournal.jpl.nasa.gov/catalog/PIA22436

1991 Southern California origin-destination survey : summary findings

DOT National Transportation Integrated Search

1993-02-01

This report presents the findings from the 1991 Southern California Origin-Destination Survey. The survey was coordinated and managed by the Southern California Association of Governments (SCAG) for the following five agencies representing their resp...

Taking the (southern) waters: science, slavery, and nationalism at the Virginia springs.

PubMed

LaFauci, Lauren E

2011-04-01

'Taking the (southern) waters' argues that, in the pre-Civil War period, the space of Virginia's mineral water resorts and the philosophy of southern hydropathic medicine enabled--indeed, fostered--white southerners' constructions of a 'nationalist,' pro-slavery ideology. In the first half of the paper, the author explains how white southern health-seekers came to view the springs region as a medicinal resource peculiarly designed for the healing of southern diseases and for the restoration of white southern constitutions; in the second half, she shows how physical and social aspects of the resorts, such as architectural choices and political events, supported and encouraged pro-slavery ideologies. Taken together, these medical-social analyses reveal how elite white southerners in the antebellum period came to associate the health of their peculiarly 'southern' bodies with the future health of an independent southern nation, one that elided black bodily presence at the same time that its social structures and scientific apparatuses relied upon enslaved black labor.

Insect and Disease Pests of Southern Hardwoods

Treesearch

L. P. Abrahamson; F. I. McCracken

1971-01-01

Insects and diseases seldom kill southern hardwood trees in managed stands, but they cause major economic losses by lowering wood quality and reducing tree growth. In discussing the most important insects and diseases of southern hardwoods, let us consider first those that attack natural hardwood stands and then those associated with plantation culture.

Southern Forest Resource Assessment - Summary of Findings

Treesearch

David N. Wear; John G. Greis

2002-01-01

The Southern Forest Resource Assessment (SFRA) was initiated in spring 1999 to address broad questions concerning the status, trends, and likely future of southern forests. A descriptive assessment such as SFRA can be used to highlight the major dynamics and uncertainties at play within a region's forested ecosystems, thereby focusing public discourse. Because...

Proceedings of the 2001 Southern Mensurationists' Conference

Treesearch

Paul F. Doruska; Don C. Bragg

2000-01-01

The 2001 Southern Mensurationists' Conference was held in scenic Chattanooga, Tennessee at the historical Chattanooga Choo Choo Holiday Inn. This conference was the latest in the series of annual gatherings of southern biometricians, and attracted speakers and participants from Texas to Virginia. A variety of papers were presented and the quaint atmosphere, as...

Distribution and phenology of ixodid ticks in southern Zambia.

PubMed

Speybroeck, N; Madder, M; Van Den Bossche, P; Mtambo, J; Berkvens, N; Chaka, G; Mulumba, M; Brandt, J; Tirry, L; Berkvens, D

2002-12-01

Distribution data for epidemiologically important ticks (Acari: Ixodidae) in the Southern Province of Zambia, one of the main cattle areas of the country, are presented. Boophilus microplus (Canestrini) was not recorded in southern Zambia, whereas Boophilus decoloratus (Koch) is present throughout the area. New distribution patterns for less economically important ixodid ticks are also discussed. Southern Zambia is a transition zone because it is the most northern area in Africa where mixed Rhipicephalus appendiculatus Neumann and Rhipicephalus zambeziensis Walker, Norval & Corwin populations were reported. Although a second generation of adult R. appendiculatus/R. zamnbeziensis was encountered, simulations indicated that this phenomenon is very rare in southern Zambia, mainly because of the colder temperatures during the early dry season and lower rainfall. These simulations were supported by a development trial under experimental conditions. Tick body size measurements showed that southern Zambian ticks are larger than eastern Zambian R. appendiculatus. It is hypothesized that body size is related to diapausing intensity in this species. The epidemiological consequences are that a different approach to control Theileria parva (Theiler) (Piroplasmida: Theileriidae) and other tick-borne diseases is needed in southern Zambia, compared to the one adopted in eastern Zambia.

A numerical investigation of the Southern Gyre using ROMS

NASA Astrophysics Data System (ADS)

Gamoyo, Majambo; Reason, Chris J. C.; Collins, Charine

2017-05-01

A numerical model (The Regional Ocean Modelling System-ROMS), configured over the western Indian Ocean and driven by monthly climatology winds and heat fluxes, is applied to examine the Southern Gyre in the Somali Current system during the Southwest Monsoon. Despite the Southern Gyre playing a role in transporting water masses and other properties northwards across the equator, it has not been much studied. The model results indicate that the Southern Gyre appears in early June in the upper ocean as a result of instability in the northward flowing Somali Current. The arrival of downwelling Rossby wave energy at the East African coast intensifies the recirculation of the Southern Gyre and causes its northward movement. The Southern Gyre is characterized as a shallow feature which deepens from 100 m in June to 300 m in July-August. The average spatial scale of the gyre is about 400 km with subsequent development of positive vorticity bursts which are identified as potential contributors to the decay of the Southern Gyre. Cool and fresh waters observed in the gyre resulted from advection via the South Equatorial Current and then through the Somali Current (SC).

Investigation of Pokemon-regulated proteins in hepatocellular carcinoma using mass spectrometry-based multiplex quantitative proteomics.

PubMed

Bi, Xin; Jin, Yibao; Gao, Xiang; Liu, Feng; Gao, Dan; Jiang, Yuyang; Liu, Hongxia

2013-01-01

Pokemon is a transcription regulator involved in embryonic development, cellular differentiation and oncogenesis. It is aberrantly overexpressed in multiple human cancers including Hepatocellular carcinoma (HCC) and is considered as a promising biomarker for HCC. In this work, the isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics strategy was used to investigate the proteomic profile associated with Pokemon in human HCC cell line QGY7703 and human hepatocyte line HL7702. Samples were labeled with four-plex iTRAQ reagents followed by two-dimensional liquid chromatography coupled with tandem mass spectrometry analysis. A total of 24 differentially expressed proteins were selected as significant. Nine proteins were potentially up-regulated by Pokemon while 15 proteins were potentially down-regulated and many proteins were previously identified as potential biomarkers for HCC. Gene ontology (GO) term enrichment revealed that the listed proteins were mainly involved in DNA metabolism and biosynthesis process. The changes of glucose-6-phosphate 1-dehydrogenase (G6PD, up-regulated) and ribonucleoside-diphosphate reductase large sub-unit (RIM1, down-regulated) were validated by Western blotting analysis and denoted as Pokemon's function of oncogenesis. We also found that Pokemon potentially repressed the expression of highly clustered proteins (MCM3, MCM5, MCM6, MCM7) which played key roles in promoting DNA replication. Altogether, our results may help better understand the role of Pokemon in HCC and promote the clinical applications.

A multiplexed quantitative proteomics approach for investigating protein expression in the developing central nervous system.

PubMed

Orme, Rowan P; Gates, Monte A; Fricker-Gates, Rosemary A

2010-08-15

Cell transplantation using stem cell-derived neurons is commonly viewed as a candidate therapy for neurodegenerative diseases. However, methods for differentiating stem cells into homogenous populations of neurons suitable for transplant remain elusive. This suggests that there are as yet unknown signalling factors working in vivo to specify neuronal cell fate during development. These factors could be manipulated to better differentiate stem cells into neural populations useful for therapeutic transplantation. Here a quantitative proteomics approach is described for investigating cell signalling in the developing central nervous system (CNS), using the embryonic ventral mesencephalon as a model. Briefly, total protein was extracted from embryonic ventral midbrain tissue before, during and after the birth of dopaminergic neurons, and digested using trypsin. Two-dimensional liquid chromatography, coupled with tandem mass spectrometry, was then used to identify proteins from the tryptic peptides. Isobaric tagging for relative and absolute quantification (iTRAQ) reagents were used to label the tryptic peptides and facilitate relative quantitative analysis. The success of the experiment was confirmed by the identification of proteins known to be expressed in the developing ventral midbrain, as well as by Western blotting, and immunolabelling of embryonic tissue sections. This method of protein discovery improves upon previous attempts to identify novel signalling factors through microarray analysis. Importantly, the methods described here could be applied to virtually any aspect of development. (c) 2010 Elsevier B.V. All rights reserved.

Physical oceanography and tracer chemistry of the southern ocean

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

Not Available

This report considers technical and scientific developments and research questions in studies of the Southern Ocean since its predecessor, /open quotes/Southern Ocean Dynamics--A Strategy for Scientific Exploration 1973-1983/close quotes/ was published. The summary lists key research questions in Southern Ocean oceanography. Chapter 1 describes how Southern Ocean research has evolved to provide the basis for timely research toward more directed objectives. Chapter 2 recommends four research programs, encompassing many of the specific recommendations that follow. Appendix A provides the scientific background and Reference/Bibliography list for this report for: on air-sea-ice interaction; the Antarctic Circumpolar Current; water mass conversion; chemical tracermore » oceanography; and numerical modeling of the Southern Ocean. Appendix B describes the satellite-based observation systems expected to be active during the next decade. Appendix C is a list of relevant reports published during 1981-1987. 146 refs.« less

Isolation of High-Molecular-Weight DNA from Mammalian Tissues Using Proteinase K and Phenol.

PubMed

Green, Michael R; Sambrook, Joseph

2017-03-01

This procedure is the method of choice for purification of genomic DNA from mammalian tissues when large amounts of DNA are required, for example, for Southern blotting. © 2017 Cold Spring Harbor Laboratory Press.

Progressive approach to eruption at Campi Flegrei caldera in southern Italy

PubMed Central

Kilburn, Christopher R.J.; De Natale, Giuseppe; Carlino, Stefano

2017-01-01

Unrest at large calderas rarely ends in eruption, encouraging vulnerable communities to perceive emergency warnings of volcanic activity as false alarms. A classic example is the Campi Flegrei caldera in southern Italy, where three episodes of major uplift since 1950 have raised its central district by about 3 m without an eruption. Individual episodes have conventionally been treated as independent events, so that only data from an ongoing episode are considered pertinent to evaluating eruptive potential. An implicit assumption is that the crust relaxes accumulated stress after each episode. Here we apply a new model of elastic-brittle failure to test the alternative view that successive episodes promote a long-term accumulation of stress in the crust. The results provide the first quantitative evidence that Campi Flegrei is evolving towards conditions more favourable to eruption and identify field tests for predictions on how the caldera will behave during future unrest. PMID:28504261

Quantitative Proteomic Analysis of Duck Ovarian Follicles Infected with Duck Tembusu Virus by Label-Free LC-MS

PubMed Central

Han, Kaikai; Zhao, Dongmin; Liu, Yuzhuo; Liu, Qingtao; Huang, Xinmei; Yang, Jing; An, Fengjiao; Li, Yin

2016-01-01

Duck Tembusu virus (DTMUV) is a newly emerging pathogenic flavivirus that has caused massive economic losses to the duck industry in China. DTMUV infection mainly results in significant decreases in egg production in egg-laying ducks within 1–2 weeks post infection. However, information on the comparative protein expression of host tissues in response to DTMUV infection is limited. In the present study, the cellular protein response to DTMUV infection in duck ovarian follicles was analyzed using nano-flow high-performance liquid chromatography-electrospray tandem mass spectrometry. Quantitative proteomic analysis revealed 131 differentially expressed proteins, among which 53 were up regulated and 78 were down regulated. The identified proteins were involved in the regulation of essential processes such as cellular structure and integrity, RNA processing, protein biosynthesis and modification, vesicle transport, signal transduction, and mitochondrial pathway. Some selected proteins that were found to be regulated in DTMUV-infected tissues were screened by quantitative real-time PCR to examine their regulation at the transcriptional level, western blot analysis was used to validate the changes of some selected proteins on translational level. To our knowledge, this study is the first to analyze the proteomic changes in duck ovarian follicles following DTMUV infection. The protein-related information obtained in this study may be useful to understand the host response to DTMUV infection and the inherent mechanism of DTMUV replication and pathogenicity. PMID:27066001

Interannual variability of the frontal activity in the Southern Hemisphere: relationship with atmospheric circulation and precipitation over southern South America

NASA Astrophysics Data System (ADS)

Blázquez, Josefina; Solman, Silvina A.

2017-04-01

The interannual variability of the frontal activity over the western Southern Hemisphere and its linkage with the variability of the atmospheric circulation and precipitation over southern South America is studied. The analysis is focused on the austral winter and spring seasons. The frontal activity is represented by an index defined as the product between the horizontal gradient of temperature and the relative vorticity at 850 hPa (FI) and is computed from the ERA Interim and NCEP2 reanalysis. For the two seasons the main mode of variability of FI, as depicted by the first Empirical Orthogonal Function, presents centres of action located in the southern part of the western Southern Hemisphere. This pattern is present in the two reanalysis datasets. The correlation coefficients between the principal component of the leading mode of FI and the two main modes of the 500 hPa geopotential height indicate that both the ENSO-mode and the SAM modulate the leading pattern of FI in winter while during the spring season the ENSO-mode controls the FI variability. The variability of the FI has a robust influence on the interannual variability of precipitation over southern South America and adjacent oceans. Over the continent, it was found that the pattern of precipitation anomalies associated with the variability of the FI depicts significant signals over southeastern South America (SESA), centre and south of Chile for winter and over SESA and southeastern Brazil for spring and agrees with the pattern of the leading mode of precipitation variability over southern South America.

A combined RT-PCR and dot-blot hybridization method reveals the coexistence of SJNNV and RGNNV betanodavirus genotypes in wild meagre (Argyrosomus regius).

PubMed

Lopez-Jimena, B; Cherif, N; Garcia-Rosado, E; Infante, C; Cano, I; Castro, D; Hammami, S; Borrego, J J; Alonso, M C

2010-10-01

To detect the possible coexistence of striped jack nervous necrosis virus (SJNNV) and red-spotted grouper nervous necrosis virus (RGNNV) genotypes in a single fish, a methodology based on the combination of PCR amplification and blot hybridization has been developed and applied in this study. The degenerate primers designed for the PCR procedure target the T4 region within the capsid gene, resulting in the amplification of both genotypes. The subsequent hybridization of these amplification products with two different specific digoxigenin-labelled probes resulted in the identification of both genotypes separately. The application of the RT-PCR protocol to analyse blood samples from asymptomatic wild meagre (Argyrosomus regius) specimens has shown a 46.87% of viral nervous necrosis virus carriers. The combination of RT-PCR and blot hybridization increases the detection rate up to 90.62%, and, in addition, it has shown the coexistence of both genotypes in 18 out of the 32 specimens analysed (56.25%). This study reports the coexistence of betanodaviruses belonging to two different genotypes (SJNNV and RGNNV) in wild fish specimens. This is the first report demonstrating the presence of SJNNV and RGNNV genotypes in the same specimen. This study also demonstrates a carrier state in this fish species for the first time. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.

Absenteeism due to mental disorders in health professionals at a hospital in southern Brazil.

PubMed

Santana, Leni de Lima; Sarquis, Leila Maria Mansano; Brey, Christiane; Miranda, Fernanda Moura D'Almeida; Felli, Vanda Elisa Andres

2016-03-01

Objective To describe the health profile of mental and behavioral disorders in health professionals at a teaching hospital in southern Brazil. Methods This was a quantitative, retrospective cross-sectional epidemiological study whose data were collected through institutional documents used to feed the Health Monitoring System for Nursing Professionals and involved all absenteeism occurred in 2011. Results We found 55 records of absenteeism due to mental and behavioral disorders, a total of 317 days absent. Nursing technicians were the most absentee, with 29.09% of the records. The intensive care unit represented the sector with the highest number of days absent, 81%, and depressive episodes were the most frequent, representing 52.72% of mental disorders. Conclusion The results showed that mental disorders in health professionals are a cause for concern and urgently need intervention.

Cone Analysis of Southern Pines - A Guidebook

Treesearch

D.L. Bramlett; E.W. Belcher; G.L. DeBarr; G.D. Hertel; Robert P. Karrfalt; C.W. Lantz; T. Miller; K.D. Ware; H.O. Yates

1977-01-01

Southern pine tree improvement programs require an ample supply of improved seeds, but productron from southern pine seed orchards has often been disappointing. If high productron is to be malntained yields must be monitored and causes of seed losses must be identified. Techniques for determining seed efficiency were first used for red pine, Pinus resinosa...

Southern Women's Literature: A Process of Discovery.

ERIC Educational Resources Information Center

Meese, Elizabeth A.

Teachers and students worked together at the University of Alabama to evolve a literature course that dealt with the Southern regional character and culture as manifested in the writings of women. The course began with an orientation to the region, with emphasis on the images and stereotypes of Southern women. Literary material included in the…

A new source of cytoplasmic male sterility in pearl millet: RFLP analysis of mitochondrial DNA.

PubMed

Sujata, V; Sivaramakrishnan, S; Rai, K N; Seetha, K

1994-06-01

A new source of cytoplasmic male sterility (cms) in pearl millet (Pennisetum glaucum (L.) R.Br.) derived from a half-sib progeny of the Early Gene Pool (EGP 261) and used in a male-sterile line, ICMA 90111, was compared with other known cms sources for RFLP of mitochondrial (mt) DNA. Southern blot hybridization of mtDNA from ICMA 90111 digested with several restriction enzymes and probed with homologous mtDNA clones from pearl millet and heterologous gene clones from maize and wheat revealed the RFLP patterns of ICMA 90111 distinct from others studied so far. The dendrogram of male-sterile lines constructed from the Southern blot hybridization patterns indicated that ICMA 90111 represents a separate group. Our results suggest that this source of cms is unique in several respects.

Southern California Edison PDF

EPA Pesticide Factsheets

This Southern California Edison report describes an evaluation of SCE’s Retail Appliance Recycling Program (Retail ARP) trial that was initiated in late October 2010 and completed in September of 2011.

Recent Precipitation Trends Over the Southern Ocean in Relation to Oceanic Freshening Near Antarctica

NASA Astrophysics Data System (ADS)

Cullather, R. I.; Jacobs, S. S.; Giulivi, C. F.; Leonard, K. C.; Stammerjohn, S. E.

2008-12-01

Quantitative assessments of large-scale precipitation over the world's oceanic regions are problematic, particularly for significant regions of the data-sparse Southern Hemisphere. Available data sets are based on the assimilation of land-based measurements, satellite radiance values, numerical weather forecast models, or some combination of the three. In this study we examine several products that cover most or all of the satellite era 1979-2007 over the Southern Ocean and surrounding mid-latitudes to 45°S. These include CMAP, the NCEP Reanalysis II, ERA-40, GPCP version 2, and the Japanese Re-analysis. Averaged fields from these data show large discrepancies in the mean spatial depiction and the annual cycle. Comparisons with unique in situ snowfall measurements and satellite-derived accumulation on sea ice are presented. The available record of oceanographic measurements in the Ross Sea indicates that salinity below 200 m in the Ross Sea has decreased by 0.03 per decade since 1958, with the highest (lowest) values in 1967 (2000). The fields examined here suggest that precipitation is likely not directly influencing the oceanic freshening observed in the Ross Sea, or in other coastal seas adjacent to Antarctica. The salinity anomaly is consistent with increasing attrition of continental ice, but places a heavy demand on the melt rate. Potential contributions to oceanic freshening from changes in sea ice extent, transport, and thickness are discussed.

The reinvigoration of the Southern Ocean carbon sink.

PubMed

Landschützer, Peter; Gruber, Nicolas; Haumann, F Alexander; Rödenbeck, Christian; Bakker, Dorothee C E; van Heuven, Steven; Hoppema, Mario; Metzl, Nicolas; Sweeney, Colm; Takahashi, Taro; Tilbrook, Bronte; Wanninkhof, Rik

2015-09-11

Several studies have suggested that the carbon sink in the Southern Ocean-the ocean's strongest region for the uptake of anthropogenic CO2 -has weakened in recent decades. We demonstrated, on the basis of multidecadal analyses of surface ocean CO2 observations, that this weakening trend stopped around 2002, and by 2012, the Southern Ocean had regained its expected strength based on the growth of atmospheric CO2. All three Southern Ocean sectors have contributed to this reinvigoration of the carbon sink, yet differences in the processes between sectors exist, related to a tendency toward a zonally more asymmetric atmospheric circulation. The large decadal variations in the Southern Ocean carbon sink suggest a rather dynamic ocean carbon cycle that varies more in time than previously recognized. Copyright © 2015, American Association for the Advancement of Science.

Emerging zoonoses in the southern United States: toxocariasis, bovine tuberculosis and southern tick-associated rash illness.

PubMed

Clinton, Rachel M; Carabin, Hélène; Little, Susan E

2010-09-01

The majority of emerging diseases in humans have been linked to zoonotic pathogens originating in domestic animals or wildlife. This is a public health concern because zoonotic infections affect several aspects of the society. The complex interactions among pathogen, host and environment also pose challenges in estimating the true burden of those infections. However, the recent development of new molecular diagnostic tools has allowed for better diagnosis of zoonotic infections. This review focuses on 3 emerging zoonoses, namely toxocariasis, bovine tuberculosis and southern tick-associated rash illness, and demonstrates that these infections may be more prevalent in the southern United States than previously recognized. This review places special emphasis on the recent epidemiologic trends, intra/interspecies transmission and clinical features of each of these zoonoses. In addition, treatment and prevention for each zoonotic pathogen are discussed. Clinicians working in the southern United States should be aware of the presence of those zoonotic infections.

An overview of the Southern Nevada Agency Partnership science and research synthesis: Chapter 1 in The Southern Nevada Agency Partnership science and research synthesis: science to support land management in southern Nevada

USGS Publications Warehouse

Chambers, Jeanne C.; Brooks, Matthew L.; Turner, Kent; Raish, Carol B.; Ostoja, Steven M.

2013-01-01

Maintaining and restoring the diverse ecosystems and resources that occur in southern Nevada in the face of rapid socio-economic and ecological change presents numerous challenged to Federal land managers. Rapid population growth since the 1980s, the land uses associated with that growth, and the interactions of those uses with the generally dry and highly variable climate result in numerous stresses to ecosystems, species, and cultural resource. In addition, climate models predict that the rate of temperature increase and, thus, changes in ecological processes, will be highest for ecosystems like the Mojave Desert. The Southern Nevada Agency Partnership (SNAP; http:www.SNAP.gov) was established in 1999 to address common issues pertaining to public lands in southern Nevada. Partners include the Bureau of Land Management, National Park Service, U.S. Fish and Wildlife Service, and USDA Forest Service and they work with each other, the local community, and other partners. SNAP agencies manage more than seven million acres of public lands in southern Nevada (95% of the land area). Federal land includes two national recreation areas, two national conservation area, four national wildlife refuges, 18 congressionally designated wilderness areas, five wilderness study areas, and 22 areas of critical environmental concern. The partnership's activities are mainly centered in Southern Nevada's Clark County (fig. 1.1), but lands managed by SNAP partner agencies also include portions of Lake Mead National Recreation Area in Mohave County, Arizona, U.S. Fish and Wildlife Service, and USDA Forest Service-managed lands in Lincoln and Nye Counties, Nevada, and all lands and activities managed by the Southern Nevada District Office of the Bureau of Land Management. These lands encompass nine distinct ecosystem types (fig. 1.2), support multiple species of management concern an 17 listed species, and are rich in cultural and historic resource. This introductory executive summary

The epidemiology of tick-borne haemoparasites as determined by the reverse line blot hybridization assay in an intensively studied cohort of calves in western Kenya

PubMed Central

Njiiri, Nyawira E.; Bronsvoort, B. Mark deC.; Collins, Nicola E.; Steyn, Helena C.; Troskie, Milana; Vorster, Ilse; Thumbi, S.M.; Sibeko, Kgomotso P.; Jennings, Amy; van Wyk, Ilana Conradie; Mbole-Kariuki, Mary; Kiara, Henry; Poole, E. Jane; Hanotte, Olivier; Coetzer, Koos; Oosthuizen, Marinda C.; Woolhouse, Mark; Toye, Philip

2015-01-01

The development of sensitive surveillance technologies using PCR-based detection of microbial DNA, such as the reverse line blot assay, can facilitate the gathering of epidemiological information on tick-borne diseases, which continue to hamper the productivity of livestock in many parts of Africa and elsewhere. We have employed a reverse line blot assay to detect the prevalence of tick-borne parasites in an intensively studied cohort of indigenous calves in western Kenya. The calves were recruited close to birth and monitored for the presence of infectious disease for up to 51 weeks. The final visit samples from 453 calves which survived for the study period were analyzed by RLB. The results indicated high prevalences of Theileria mutans (71.6%), T. velifera (62.8%), Anaplasma sp. Omatjenne (42.7%), A. bovis (39.9%), Theileria sp. (sable) (32.7%), T. parva (12.9%) and T. taurotragi (8.5%), with minor occurrences of eight other haemoparasites. The unexpectedly low prevalence of the pathogenic species Ehrlichia ruminantium was confirmed by a species-specific PCR targeting the pCS20 gene region. Coinfection analyses of the seven most prevalent haemoparasites indicated that they were present as coinfections in over 90% of the cases. The analyses revealed significant associations between several of the Theileria parasites, in particular T. velifera with Theileria sp. sable and T. mutans, and T. parva with T. taurotragi. There was very little coinfection of the two most common Anaplasma species, although they were commonly detected as coinfections with the Theileria parasites. The comparison of reverse line blot and serological results for four haemoparasites (T. parva, T. mutans, A. marginale and B. bigemina) indicated that, except for the mostly benign T. mutans, indigenous cattle seem capable of clearing infections of the three other, pathogenic parasites to below detectable levels. Although the study site was located across four agroecological zones, there was

Alluvial Scrub Vegetation in Coastal Southern California

Treesearch

Ted L. Hanes; Richard D. Friesen; Kathy Keane

1989-01-01

Certain floodplain systems in southern California sustain a unique scrub vegetation rather than riparian woodlands due to a lack of perennial water. Alluvial scrub occurs on outwash fans and riverine deposits along the coastal side of major mountains of southern California. This vegetation type is adapted to severe floods and erosion, nutrient-poor substrates, and the...

Mechanical Control of Southern Pine Beetle Infestations

Treesearch

Ronald F. Billings

2011-01-01

Periodic outbreaks of the southern pine beetle (SPB) may affect thousands of acres of commercial pine forests in the Southeastern United States, Mexico, and Central America. Accordingly, this species is the target of more aggressive and effective suppression programs than any other bark beetle pest in the world. The strategy for controlling the southern pine beetle...

Syntectonic remagnetization in the southern Methow block: Resolving large displacements in the southern Canadian Cordillera

USGS Publications Warehouse

Enkin, R.J.; Mahoney, J.B.; Baker, J.; Kiessling, M.; Haugerud, R.A.

2002-01-01

The Upper Cretaceous Ventura Member of the Goat Wall unit in the southern Methow block of southern British Columbia and northern Washington State holds a syntectonic magnetization. Eight new sites from Manning Park in British Columbia give a mean direction of D = 27.5??, I = 60.1??, k = 304.7, ??95 = 3.2?? after optimal partial tilt correction. Of five groups of bedded sites from farther south in the basin reported by Bazard et al. [1990], four have a syntectonic remanence with a direction similar to what we observe. The exception is one group which has optimal concentration of remanence directions on >100% untilting and an abherent direction which must be rejected. Combining the accepted sites, the optimal differential syntilting direction is D = 11.8??, I = 61.5??, k = 39.3, ??95 = 3.4?? (N = 47), giving a mean pole of 79.8??N, 359.2??E, K = 19.5, and A95 = 4.8??. The age of the remagnetization is constrained to be between 88 and 80 Ma. Compared to cratonic North America, this result indicates that the southern Methow block was displaced from the south by 1800 ?? 500 km, meaning it lay south of the Sierra Nevada subduction zone but well north of other paleomagnetically constrained Cretaceous rock units from the Insular superterrane, including correlative strata of the Mount Tatlow area in the northern Methow block. Among several possibilities to reconcile this discrepancy, the most plausible has the whole Methow block translated coherently but with the southern Methow block strata remagnetized during transit.

75 FR 52715 - Southern Arizona Resource Advisory Committee

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-27

... DEPARTMENT OF AGRICULTURE Forest Service Southern Arizona Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Southern Arizona Resource Advisory... held at the National Advanced Fire and Resource Institute (NAFRI) at 3265 E. Universal Way, Tucson...

75 FR 60066 - Southern Arizona Resource Advisory Committee

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-29

... DEPARTMENT OF AGRICULTURE Forest Service Southern Arizona Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Southern Arizona Resource Advisory... at the Tucson Interagency Fire Center, 2646 E. Commerce Center Place, Tucson, AZ 85706. Send written...

DIET OF THE SOUTHERN TOAD (BUFO TERRESTRIS) FROM THE SOUTHERN EVERGLADES

EPA Science Inventory

We examined the diet of a February-May sample of the southern toad (Bufo terrestris) from the Everglades National Park. Above the familial level, 13 taxa were consumed, but ants (Hymenoptera) and beetles (Coleoptera) were consumed most by, and in the greatest number of sto...

Southern Ocean Deep-Convection as a Driver of Centennial-to-Millennial-Scale Climate Variability at Southern High Latitudes

NASA Astrophysics Data System (ADS)

Pedro, J. B.; Martin, T.; Steig, E. J.; Jochum, M.; Park, W.; Rasmussen, S.

2015-12-01

Antarctic Isotope Maxima (AIM) are centennial-to-millennial scale warming events observed in Antarctic ice core records from the last glacial period and deglaciation. Mounting evidence links AIM events to parallel variations in atmospheric CO2, Southern Ocean (SO) sea surface temperatures and Antarctic Bottom Water production. According to the prevailing view, AIM events are forced from the North Atlantic by melt-water discharge from ice sheets suppressing the production of North Atlantic Deep Water and associated northward heat transport in the Atlantic. However observations and model studies increasingly suggest that melt-water fluxes have the wrong timing to be invoked as such a trigger. Here, drawing on results form the Kiel Climate Model, we present an alternative hypothesis in which AIM events are forced via internal oscillations in SO deep-convection. The quasi-periodic timescale of deep-convection events is set by heat (buoyancy) accumulation at SO intermediate depths and stochastic variability in sea ice conditions and freshening at the surface. Massive heat release from the SO convective zone drives Antarctic and large-scale southern hemisphere warming via a two-stage process involving changes in the location of Southern Ocean fronts, in the strength and intensity of the Westerlies and in meridional ocean and atmospheric heat flux anomalies. The potential for AIM events to be driven by internal Southern Ocean processes and the identification of time-lags internal to the southern high latitudes challenges conventional views on the North Atlantic as the pacemaker of millennial-scale climate variability.

The genetic prehistory of southern Africa.

PubMed

Pickrell, Joseph K; Patterson, Nick; Barbieri, Chiara; Berthold, Falko; Gerlach, Linda; Güldemann, Tom; Kure, Blesswell; Mpoloka, Sununguko Wata; Nakagawa, Hirosi; Naumann, Christfried; Lipson, Mark; Loh, Po-Ru; Lachance, Joseph; Mountain, Joanna; Bustamante, Carlos D; Berger, Bonnie; Tishkoff, Sarah A; Henn, Brenna M; Stoneking, Mark; Reich, David; Pakendorf, Brigitte

2012-01-01

Southern and eastern African populations that speak non-Bantu languages with click consonants are known to harbour some of the most ancient genetic lineages in humans, but their relationships are poorly understood. Here, we report data from 23 populations analysed at over half a million single-nucleotide polymorphisms, using a genome-wide array designed for studying human history. The southern African Khoisan fall into two genetic groups, loosely corresponding to the northwestern and southeastern Kalahari, which we show separated within the last 30,000 years. We find that all individuals derive at least a few percent of their genomes from admixture with non-Khoisan populations that began ∼1,200 years ago. In addition, the East African Hadza and Sandawe derive a fraction of their ancestry from admixture with a population related to the Khoisan, supporting the hypothesis of an ancient link between southern and eastern Africa.

Connecting qualitative observation and quantitative measurement for enhancing quantitative literacy in plant anatomy course

NASA Astrophysics Data System (ADS)

Nuraeni, E.; Rahmat, A.

2018-05-01

Forming of cognitive schemes of plant anatomy concepts is performed by processing of qualitative and quantitative data obtained from microscopic observations. To enhancing student’s quantitative literacy, strategy of plant anatomy course was modified by adding the task to analyze quantitative data produced by quantitative measurement of plant anatomy guided by material course. Participant in this study was 24 biology students and 35 biology education students. Quantitative Literacy test, complex thinking in plant anatomy test and questioner used to evaluate the course. Quantitative literacy capability data was collected by quantitative literacy test with the rubric from the Association of American Colleges and Universities, Complex thinking in plant anatomy by test according to Marzano and questioner. Quantitative literacy data are categorized according to modified Rhodes and Finley categories. The results showed that quantitative literacy of biology education students is better than biology students.

A quantitative study on the mass elevation effect of the Rocky Mountains and its significance for treeline distribution

NASA Astrophysics Data System (ADS)

WANG, J.; Zhang, B.

2016-12-01

The Rocky Mountains are the highest and most extensive of all in the North America. To quantify mass elevation effect (MEE) of the Rocky Mountains, we applied meteorological station records, NCAR/NCEP free air temperature and DEM data to calculate temperature difference (ΔT) between the inner and outer Rocky Mountains, defined as the magnitude of MEE. Results show that the mean ΔT for all adopted stations was 1.8 °, with high ΔT occurring in the Southern Rocky Mountains in the Colorado State and in the basins of Southern Wyoming. The MEE of the Rocky Mountains can be modeled with three factors of mountain base elevation (MBE), latitude and hygric continentality as independent variables. The model has a high explanatory power of 68.9%, and the three factors contribute 45.65%, 36.05% and 18.03%, respectively. Especially, MBE contributed the most to MEE of both the whole and the Southern Rocky Mountains, i.e., 45.65%, and 55.21%, respectively. Moreover, we investigated the significance of MEE for treeline distribution. The treeline is always higher in the inner than in the outer mountains, with a difference of 600 m to 1300 m. This difference corresponds well to air temperature difference between the inner and outer mountain ranges. This study developed a quantitative model for the MEE of the Rocky Mountains and improves our understanding of the intra-mountain ecology and especially the high treelines in the Rocky Mountains.

Identification of Tengfu Jiangya Tablet Target Biomarkers with Quantitative Proteomic Technique

PubMed Central

Xu, Jingwen; Zhang, Shijun; Jiang, Haiqiang; Wang, Nan; Lin, Haiqing

2017-01-01

Tengfu Jiangya Tablet (TJT) is a well accepted antihypertension drug in China and its major active components were Uncaria total alkaloids and Semen Raphani soluble alkaloid. To further explore treatment effects mechanism of TJT on essential hypertension, a serum proteomic study was performed. Potential biomarkers were quantified in serum of hypertension individuals before and after taking TJT with isobaric tags for relative and absolute quantitation (iTRAQ) coupled two-dimensional liquid chromatography followed electrospray ionization-tandem mass spectrometry (2D LC-MS/MS) proteomics technique. Among 391 identified proteins with high confidence, 70 proteins were differentially expressed (fold variation criteria, >1.2 or <0.83) between two groups (39 upregulated and 31 downregulated). Combining with Gene Ontology annotation, KEGG pathway analysis, and literature retrieval, 5 proteins were chosen as key target biomarkers during TJT therapeutic process. And the alteration profiles of these 5 proteins were verified by ELISA and Western Blot. Proteins Kininogen 1 and Keratin 1 are members of Kallikrein system, while Myeloperoxidase, Serum Amyloid protein A, and Retinol binding protein 4 had been reported closely related to vascular endothelial injury. Our study discovered 5 target biomarkers of the compound Chinese medicine TJT. Secondly, this research initially revealed the antihypertension therapeutic mechanism of this drug from a brand-new aspect. PMID:28408942

Quantitative expression and localization of cysteine and aspartic proteases in human abdominal aortic aneurysms

PubMed Central

Lohoefer, Fabian; Reeps, Christian; Lipp, Christina; Rudelius, Martina; Haertl, Felix; Matevossian, Edouard; Zernecke, Alma; Eckstein, Hans-Henning; Pelisek, Jaroslav

2014-01-01

Cysteine and aspartic proteases possess high elastolytic activity and might contribute to the degradation of the abdominal aortic aneurysm (AAA) wall. The aim of this study was to analyze, in detail, the proteases (cathepsins B, D, K, L and S, and inhibitor cystatin C) found in human AAA and healthy aortic tissue samples. The vessel walls from AAA patients (n=36) and nonaneurysmal aortae (n=10) were retrieved using conventional surgical repair and autopsy methods. Serum samples from the same AAA patients and 10 healthy volunteers were also collected. Quantitative expression analyses were performed at the mRNA level using real-time reverse transcriptase-PCR (RT–PCR). Furthermore, analyses at the protein level included western blot and immunoprecipitation analyses. Cellular sources of cysteine/aspartic proteases and cystatin C were identified by immunohistochemistry (IHC). All cysteine/aspartic proteases and cystatin C were detected in the AAA and control samples. Using quantitative RT–PCR, a significant increase in expression was observed for cathepsins B (P=0.021) and L (P=0.018), compared with the controls. Cathepsin B and cystatin C were also detected in the serum of AAA patients. Using IHC, smooth muscle cells (SMCs) and macrophages were positive for all of the tested cathepsins, as well as cystatin C; in addition, the lymphocytes were mainly positive for cathepsin B, followed by cathepsins D and S. All cysteine/aspartic proteases analyzed in our study were detected in the AAA and healthy aorta. The highest expression was found in macrophages and SMCs. Consequently, cysteine/aspartic proteases might play a substantial role in AAA. PMID:24833013

Southern Pine Beetle Handbook: Southern Pine Beetles Can Kill Your Ornamental Pine

Treesearch

Robert C. Thatcher; Jack E. Coster; Thomas L. Payne

1974-01-01

Southern pine beetles are compulsive eaters. Each year in the South from Texas to Virginia the voracious insects conduct a movable feast across thousands of acres of pine forests. Most trees die soon after the beetles sink their teeth into them.

Holocene Geologic Slip Rate for the Banning Strand of the Southern San Andreas Fault near San Gorgonio Pass, Southern California

NASA Astrophysics Data System (ADS)

Gold, P. O.; Behr, W. M.; Rood, D. H.; Kendrick, K. J.; Rockwell, T. K.; Sharp, W. D.

2014-12-01

We present the first Holocene geologic slip rate for the Banning strand of the southern San Andreas Fault in southern California. The southern San Andreas Fault splays into the sub-parallel Banning and Mission Creek strands in the northwestern Coachella Valley, and although it has long been surmised that the Banning strand eventually accommodates the majority of displacement and transfers it into San Gorgonio Pass, until now it has been uncertain how slip is actually partitioned between these two fault strands. Our new slip rate measurement, critically located at the northwestern end of the Banning strand, overlaps within errors with the published rate for the southern San Andreas Fault measured at Biskra Palms Oasis. This indicates that the majority of southern San Andreas Fault displacement transfers from the southeastern Mission Creek strand northwest to the Banning strand and into San Gorgonio Pass. Our result corroborates the UCERF3 hazard model, and is consistent with most previous interpretations of how slip is partitioned between the Banning and Mission Creek fault strands. To measure this slip rate, we used B4 airborne LiDAR to identify the apex of an alluvial fan offset laterally 30 ± 5 m from its source. We calculated the depositional age of the fan using 10Be in-situ cosmogenic exposure dating of 5 cobbles and a depth profile. We calculated a most probable fan age of 4.0 +2.0/-1.6 ka (1σ) by combining the inheritance-corrected cobble ages assuming Gaussian uncertainty. However, the probability density function yielded a multi-peaked distribution, which we attribute to variable 10Be inheritance in the cobbles, so we favor the depth profile age of 2.2-3.6 ka. Combined, these measurements yield a late Holocene slip rate for the Banning strand of the southern San Andreas Fault of 11.1 +3.1/-3.3 mm/yr. This slip rate does not preclude possibility that some slip transfers north along the Mission Creek strand and the Garnet Hill fault, but it does confirm

Prevalence of Vibrio cholerae with heat-stable enterotoxin (NAG-ST) and cholera toxin genes; restriction fragment length polymorphisms of NAG-ST genes among V. cholerae O serogroups from a major shrimp production area in Thailand.

PubMed

Dalsgaard, A; Serichantalergs, O; Shimada, T; Sethabutr, O; Echeverria, P

1995-09-01

A total of 148 Vibrio cholerae isolates from a major shrimp production area in Southern Thailand were examined by colony hybridisation for genes encoding heat-stable enterotoxin (NAG-ST) and cholera toxin (CT). Only non-O1 V. cholerae strains were found to harbour NAG-ST (14 of 146) whereas no strains hybridised with the CT probe. NAG-ST-positive V. cholerae non-O1 strains were isolated from shrimp farms situated close to urban areas. Five different O serogroups were found among NAG-ST positive non-O1 strains. Southern blot and restriction endonuclease analysis of NAG-ST-positive strains revealed a high degree of genetic divergence. A total of seven classes of enterotoxin gene patterns were found with HindIII and EcoRI restriction endonucleases. Enterotoxin gene patterns correlated with O-antigen expression in 84% of isolates tested. In combination with other molecular techniques Southern blot analysis with an NAG-ST oligonucleotide probe could be useful for studying the molecular epidemiology of V. cholerae non-O1 strains.

Matrix-assisted laser desorption-ionization mass spectrometry peptide mass fingerprinting for proteome analysis: identification efficiency after on-blot or in-gel digestion with and without desalting procedures.

PubMed

Lamer, S; Jungblut, P R

2001-03-10

In theory, peptide mass fingerprinting by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower microl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration.

Southern Forest Resource Assessment and Linkages to the National RPA

Treesearch

Fredrick Cubbage; Jacek Siry; Steverson Moffat; David N. Wear; Robert Abt

1998-01-01

We developed a Southern Forest Resource Assessment Consortium (SOFAC) in 1994, which is designed to enhance our capabilities to analyze and model the southern forest and timber resources. Southern growth and yield analyses prepared for the RPA via SOFAC indicate that substantial increases in timber productivity can occur given current technology. A survey about NIPF...

Establishing a Southern Swedish Malignant Melanoma OMICS and biobank clinical capability

PubMed Central

2013-01-01

melanoma tumors, as well as reference blood standards that is used by the team members in the respective laboratories. These pilot data show biobank access and feasibility of performing quantitative proteomics in MM biobank repositories collected in southern Sweden. The scientific outcomes further strengthen the build of healthcare benefit in the complex challenges of malignant melanoma pathophysiology that is addressed by the novel personalized medicines entering the market. PMID:23445834

Establishing a Southern Swedish Malignant Melanoma OMICS and biobank clinical capability.

PubMed

Welinder, Charlotte; Jönsson, Göran; Ingvar, Christian; Lundgren, Lotta; Olsson, Håkan; Breslin, Thomas; Végvári, Akos; Laurell, Thomas; Rezeli, Melinda; Jansson, Bo; Baldetorp, Bo; Marko-Varga, György

2013-02-27

standards that is used by the team members in the respective laboratories. These pilot data show biobank access and feasibility of performing quantitative proteomics in MM biobank repositories collected in southern Sweden. The scientific outcomes further strengthen the build of healthcare benefit in the complex challenges of malignant melanoma pathophysiology that is addressed by the novel personalized medicines entering the market.

The beginnings of the Southern Child/Pediatric Neurology Society.

PubMed

Dyken, Paul Richard; Bodensteiner, John B

2015-04-01

The founding and early development of the Southern Pediatric Neurology Society was in many ways parallel to that of the Child Neurology Society. The organization started out as the Southern Child Neurology Society but the name was changed at the time of incorporation so as to avoid confusion of identity and purpose with the larger Child Neurology Society. Although there are archives of early days and the later development of the Southern Pediatric Neurology Society, the details have never been set down in a narrative explaining the events that led to the development of the organization. In this paper, we try to produce a written record of the history of the founding and early development of the Southern Pediatric Neurology Society. © The Author(s) 2014.

Southern Ocean Climate and Sea Ice Anomalies Associated with the Southern Oscillation.

NASA Astrophysics Data System (ADS)

Kwok, R.; Comiso, J. C.

2002-03-01

The anomalies in the climate and sea ice cover of the Southern Ocean and their relationships with the Southern Oscillation (SO) are investigated using a 17-yr dataset from 1982 to 1998. The polar climate anomalies are correlated with the Southern Oscillation index (SOI) and the composites of these anomalies are examined under the positive (SOI > 0), neutral (0 > SOI > 1), and negative (SOI < 1) phases of SOI. The climate dataset consists of sea level pressure, wind, surface air temperature, and sea surface temperature fields, while the sea ice dataset describes its extent, concentration, motion, and surface temperature. The analysis depicts, for the first time, the spatial variability in the relationship of the above variables with the SOI. The strongest correlation between the SOI and the polar climate anomalies are found in the Bellingshausen, Amundsen, and Ross Seas. The composite fields reveal anomalies that are organized in distinct large-scale spatial patterns with opposing polarities at the two extremes of SOI, and suggest oscillations that are closely linked to the SO. Within these sectors, positive (negative) phases of the SOI are generally associated with lower (higher) sea level pressure, cooler (warmer) surface air temperature, and cooler (warmer) sea surface temperature in these sectors. Associations between these climate anomalies and the behavior of the Antarctic sea ice cover are evident. Recent anomalies in the sea ice cover that are clearly associated with the SOI include the following: the record decrease in the sea ice extent in the Bellingshausen Sea from mid-1988 to early 1991; the relationship between Ross Sea SST and the ENSO signal, and reduced sea ice concentration in the Ross Sea; and the shortening of the ice season in the eastern Ross Sea, Amundsen Sea, far western Weddell Sea and lengthening of the ice season in the western Ross Sea, Bellinghausen Sea, and central Weddell Sea gyre during the period 1988-94. Four ENSO episodes over the

The Moving Edge: Perspectives on the Southern Wildland-Urban Interface

Treesearch

Martha C. Monroe; Alison W. Bowers; L. Annie Hermansen

2003-01-01

To better understand the wildland-urban interface across the 13 Southern States and to identify issues to be covered in the USDA Forest Service report, "Human Influences on Forest Ecosystems: The Southern Wildland-Urban Interface Assessment," 12 focus groups were conducted in 6 of the Southern States in May and June 2000. The groups were guided through a...

Resource-Sharing and Self-Reliance in Southern Africa.

ERIC Educational Resources Information Center

Lungu, Charles B. M.

1987-01-01

Discusses the need for a regional information network in southern Africa to provide policy makers with access to needed information and reduce dependence on foreign advisers. The progress of the Southern Africa Documentation and Information System is described, and future trends and possible difficulties are identified. (CLB)

Quantitative research.

PubMed

Watson, Roger

2015-04-01

This article describes the basic tenets of quantitative research. The concepts of dependent and independent variables are addressed and the concept of measurement and its associated issues, such as error, reliability and validity, are explored. Experiments and surveys – the principal research designs in quantitative research – are described and key features explained. The importance of the double-blind randomised controlled trial is emphasised, alongside the importance of longitudinal surveys, as opposed to cross-sectional surveys. Essential features of data storage are covered, with an emphasis on safe, anonymous storage. Finally, the article explores the analysis of quantitative data, considering what may be analysed and the main uses of statistics in analysis.

Southern pulpwood production, 1981

Treesearch

Cecil C. Hutchins

1982-01-01

Southern pulpwood production has remained almost constant for the past 3 years. In 1981, production totaled 54.3 million cords with softwood roundwood accounting for about 50 percent, hardwood roundwood 18 percent, and mill byproducts 32 percent.

New Zealand Southern Alps

NASA Image and Video Library

2001-06-20

This anaglyph from the MISR instrument aboard NASA Terra spacecraft shows the rugged Southern Alps extending some 650 kilometers along the western side of New Zealand South Island. 3D glasses are necessary to view this image.

Transport of Biomass Burning Emissions from Southern Africa

NASA Technical Reports Server (NTRS)

Sinha, Parikhit; Jaegle,Lyatt; Hobbs, Peter V.; Liang, Qing

2004-01-01

The transport of biomass burning emissions from southern Africa to the neighboring Atlantic and Indian Oceans during the dry season (May-October) of 2000 is characterized using ground, ozonesonde, and aircraft measurements of carbon monoxide (CO) and ozone (O3) in and around southern Africa, together with the GEOS-CHEM global model of tropospheric chemistry. The model shows a positive bias of approximately 20% for CO and a negative bias of approximately 10-25% for O3 at oceanic sites downwind of fire emissions. Near areas of active fire emissions the model shows a negative bias of approximately 60% and approximately 30% for CO and O3, respectively, likely due to the coarse spatial (2 deg. x 2.5 deg.) and temporal (monthly) resolution of the model compared to that of active fires. On average, from 1994 to 2000, approximately 60 Tg of carbon monoxide (CO) from biomass burning in southern Africa was transported eastward to the Indian Ocean across the latitude band 0 deg. -60 S during the 6 months of the dry season. Over the same time period, approximately 40 Tg of CO from southern African biomass burning was transported westward to the Atlantic Ocean over the latitudes 0 deg. -20 S during the 6-month dry season, but most of that amount was transported back eastward over higher latitudes to the south (21 deg. -60 S). Eastward transport of biomass burning emissions from southern Africa enhances CO concentrations by approximately 4- 13 ppbv per month over the southern subtropical Indian Ocean during the dry season, with peak enhancements in September. Carbon monoxide from southern African and South American biomass burning is seen in the model simulations as far away as Australia, contributing approximately 8 ppbv and approximately 12-15 ppbv CO, respectively, and thus explaining the approximately 20- 25 ppbv observed enhancement of CO over Melbourne in mid-September 2000.

The Southern Forest Futures Project: summary report

Treesearch

David N. Wear; John G. Greis

2012-01-01

The Southern Forest Futures Project provides a science-based “futuring” analysis of the forests of the 13 States of the Southeastern United States. With findings organized in a set of scenarios and using a combination of computer models and science synthesis, the authors of the Southern Forest Futures Project examine a variety of possible futures that could shape...

Southern Hemisphere rainfall variability over the past 200 years

NASA Astrophysics Data System (ADS)

Gergis, Joëlle; Henley, Benjamin J.

2017-04-01

This study presents an analysis of three palaeoclimate rainfall reconstructions from the Southern Hemisphere regions of south-eastern Australia (SEA), southern South Africa (SAF) and southern South America (SSA). We provide a first comparison of rainfall variations in these three regions over the past two centuries, with a focus on identifying synchronous wet and dry periods. Despite the uncertainties associated with the spatial and temporal limitations of the rainfall reconstructions, we find evidence of dynamically-forced climate influences. An investigation of the twentieth century relationship between regional rainfall and the large-scale climate circulation features of the Pacific, Indian and Southern Ocean regions revealed that Indo-Pacific variations of the El Niño-Southern Oscillation (ENSO) and the Indian Ocean dipole dominate rainfall variability in SEA and SAF, while the higher latitude Southern Annular Mode (SAM) exerts a greater influence in SSA. An assessment of the stability of the regional rainfall-climate circulation modes over the past two centuries revealed a number of non-stationarities, the most notable of which occurs during the early nineteenth century around 1820. This corresponds to a time when the influence of ENSO on SEA, SAF and SSA rainfall weakens and there is a strengthening of the influence of SAM. We conclude by advocating the use of long-term palaeoclimate data to estimate decadal rainfall variability for future water resource management.

Quantitative Story Telling: Initial steps towards bridging perspectives and tools for a robust nexus assessment

NASA Astrophysics Data System (ADS)

Cabello, Violeta

2017-04-01

This communication will present the advancement of an innovative analytical framework for the analysis of Water-Energy-Food-Climate Nexus termed Quantitative Story Telling (QST). The methodology is currently under development within the H2020 project MAGIC - Moving Towards Adaptive Governance in Complexity: Informing Nexus Security (www.magic-nexus.eu). The key innovation of QST is that it bridges qualitative and quantitative analytical tools into an iterative research process in which each step is built and validated in interaction with stakeholders. The qualitative analysis focusses on the identification of the narratives behind the development of relevant WEFC-Nexus policies and innovations. The quantitative engine is the Multi-Scale Analysis of Societal and Ecosystem Metabolism (MuSIASEM), a resource accounting toolkit capable of integrating multiple analytical dimensions at different scales through relational analysis. Although QST may not be labelled a data-driven but a story-driven approach, I will argue that improving models per se may not lead to an improved understanding of WEF-Nexus problems unless we are capable of generating more robust narratives to frame them. The communication will cover an introduction to MAGIC project, the basic concepts of QST and a case study focussed on agricultural production in a semi-arid region in Southern Spain. Data requirements for this case study and the limitations to find, access or estimate them will be presented alongside a reflection on the relation between analytical scales and data availability.

75 FR 77677 - Southern Nuclear Operating Company, Inc.

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-13

... NUCLEAR REGULATORY COMMISSION [Docket No. 50-366; NRC-2010-0345] Southern Nuclear Operating Company, Inc. Edwin I Hatch Nuclear Plant, Unit No. 2; Exemption 1.0 Background The Southern Nuclear...-5 which authorizes operation of the Edwin I. Hatch Nuclear Plant, Unit No. 2 (HNP-2). The license...

Southern pulpwood production, 1961

Treesearch

Vernon L. Robinson; Agnes C. Nichols

1962-01-01

Southern pulpwood production reached 24,230,728 cords in 1961--60 percent of the Nations total. Significant increases were noted in the consumption of hardwood and residues. But pine roundwood remained virtually unchanged for the third consecutive year.

Optimal Level of Expenditure to Control the Southern Pine Beetle

Treesearch

Joseph E. de Steiguer; Roy L. Hedden; John M. Pye

1987-01-01

Optimal level of expenditure to control damage to commercial timber stands by the southern pine beetle was determined by models that simulated and analyzed beetle attacks during a typical season for 11 Southern States. At a real discount rate of 4 percent, maximized net benefits for the Southern region are estimated at about $50 million; at 10 percent, more than $30...

Application of a reverse dot blot DNA-DNA hydridization method to quantify host-feeding tendencies of two sibling species in the Anopheles gambiae complex.

PubMed

Fritz, M L; Miller, J R; Bayoh, M N; Vulule, J M; Landgraf, J R; Walker, E D

2013-12-01

A DNA-DNA hybridization method, reverse dot blot analysis (RDBA), was used to identify Anopheles gambiae s.s. and Anopheles arabiensis (Diptera: Culicidae) hosts. Of 299 blood-fed and semi-gravid An. gambiae s.l. collected from Kisian, Kenya, 244 individuals were identifiable to species; of these, 69.5% were An. arabiensis and 29.5% were An. gambiae s.s. Host identifications with RDBA were comparable with those of conventional polymerase chain reaction (PCR) followed by direct sequencing of amplicons of the vertebrate mitochondrial cytochrome b gene. Of the 174 amplicon-producing samples used to compare these two methods, 147 were identifiable by direct sequencing and 139 of these were identifiable by RDBA. Anopheles arabiensis bloodmeals were mostly (94.6%) bovine in origin, whereas An. gambiae s.s. fed upon humans more than 91.8% of the time. Tests by RDBA detected that two of 112 An. arabiensis contained blood from more than one host species, whereas PCR and direct sequencing did not. Recent use of insecticide-treated bednets in Kisian is likely to have caused the shift in the dominant vector species from An. gambiae s.s. to An. arabiensis. Reverse dot blot analysis provides an opportunity to study changes in host-feeding by members of the An. gambiae complex in response to the broadening distribution of vector control measures targeting host-selection behaviours. © 2013 The Royal Entomological Society.

Paleolakes in the Gobi region of southern Mongolia

NASA Astrophysics Data System (ADS)

Lehmkuhl, Frank; Grunert, Jörg; Hülle, Daniela; Batkhishig, Ochirbat; Stauch, Georg

2018-01-01

Numerous lakes and remnants of paleolakes exist in western and southern Mongolia. For six basins in the area, detailed geomorphological maps were compiled, based on extensive field studies and remote sensing datasets. Several phases of high and low lake levels were reconstructed and dated by radiocarbon and optically stimulated luminescence. During the marine isotope stage (MIS) 6 lakes in southern and western Mongolia mostly disappeared. In contrast, large paleolakes existed during the last interglacial (MIS 5e) and lasted probably until the beginning of the last glacial. These huge lakes were caused by a strong East Asian summer monsoon, which reached southern and even western Mongolia. During the MIS 3 the monsoon was considerably weaker and most of the lakes were relatively small or even disappeared. Higher lake levels of this period were only recorded at the Orog Nuur. However, at this time the lake was fed by glacial melt water from the Khangai Mountains. The MIS 2 was again a very dry period. The previously supposed phase of synchronous high lake levels and glaciations in southern and western Mongolia is not supported by the data presented here. During the Holocene, lakes in the western and southern part of the study area evolved differently. Early Holocene high lake levels were reconstructed for the western lakes, while most of the southern lakes had highest lake levels in the mid-Holocene. These differences can be attributed to different moisture bearing atmospheric systems. In the late Holocene lake levels were generally low and in the last 50 years most lakes completely disappeared due to a strong human usage of the water resources.

Accretion of southern Alaska

USGS Publications Warehouse

Hillhouse, J.W.

1987-01-01

Paleomagnetic data from southern Alaska indicate that the Wrangellia and Peninsular terranes collided with central Alaska probably by 65 Ma ago and certainly no later than 55 Ma ago. The accretion of these terranes to the mainland was followed by the arrival of the Ghost Rocks volcanic assemblage at the southern margin of Kodiak Island. Poleward movement of these terranes can be explained by rapid motion of the Kula oceanic plate, mainly from 85 to 43 Ma ago, according to recent reconstructions derived from the hot-spot reference frame. After accretion, much of southwestern Alaska underwent a counterclockwise rotation of about 50 ?? as indicated by paleomagnetic poles from volcanic rocks of Late Cretaceous and Early Tertiary age. Compression between North America and Asia during opening of the North Atlantic (68-44 Ma ago) may account for the rotation. ?? 1987.

DNA methylation and soy phytoestrogens: quantitative study in DU-145 and PC-3 human prostate cancer cell lines.

PubMed

Adjakly, Mawussi; Bosviel, Rémy; Rabiau, Nadège; Boiteux, Jean-Paul; Bignon, Yves-Jean; Guy, Laurent; Bernard-Gallon, Dominique

2011-12-01

DNA hypermethylation is an epigenetic mechanism which induces silencing of tumor-suppressor genes in prostate cancer. Many studies have reported that specific components of food plants like soy phytoestrogens may have protective effects against prostate carcinogenesis or progression. Genistein and daidzein, the major phytoestrogens, have been reported to have the ability to reverse DNA hypermethylation in cancer cell lines. The aim of this study was to investigate the potential demethylating effects of these two soy compounds on BRCA1, GSTP1, EPHB2 and BRCA2 promoter genes. Prostate cell lines DU-145 and PC-3 were treated with genistein 40 µM, daidzein 110 µM, budesonide (methylating agent) 2 µM and 5-azacytidine (demethylating agent) 2 µM. In these two human prostate cancer cell lines we performed methylation quantification by using Methyl Profiler DNA methylation analysis. This technique is based on a methylation-specific digestion followed by quantitative PCR. We analyzed the corresponding protein expression by western blotting. Soy phytoestrogens induced a demethylation of all promoter regions studied except for BRCA2, which is not methylated in control cell lines. An increase in their protein expression was also demonstrated by western blot analysis and corroborated the potential demethylating effect of soy phytoestrogens. This study showed that the soy phytoestrogens, genistein and daidzein, induce a decrease of methylation of BRCA1, GSTP1 and EPHB2 promoters. Therefore, soy phytoestrogens may have a protective effect on prostate cancer. However, more studies are needed in order to understand the mechanism by which genistein and daidzein have an inhibiting action on DNA methylation.

40 CFR 81.158 - Southern Wisconsin Intrastate Air Quality Control Region.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 17 2010-07-01 2010-07-01 false Southern Wisconsin Intrastate Air... Air Quality Control Regions § 81.158 Southern Wisconsin Intrastate Air Quality Control Region. The Southern Wisconsin Intrastate Air Quality Control Region consists of the territorial area encompassed by...

40 CFR 81.158 - Southern Wisconsin Intrastate Air Quality Control Region.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 17 2011-07-01 2011-07-01 false Southern Wisconsin Intrastate Air... Air Quality Control Regions § 81.158 Southern Wisconsin Intrastate Air Quality Control Region. The Southern Wisconsin Intrastate Air Quality Control Region consists of the territorial area encompassed by...

40 CFR 81.158 - Southern Wisconsin Intrastate Air Quality Control Region.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 18 2013-07-01 2013-07-01 false Southern Wisconsin Intrastate Air... Air Quality Control Regions § 81.158 Southern Wisconsin Intrastate Air Quality Control Region. The Southern Wisconsin Intrastate Air Quality Control Region consists of the territorial area encompassed by...

40 CFR 81.158 - Southern Wisconsin Intrastate Air Quality Control Region.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 18 2014-07-01 2014-07-01 false Southern Wisconsin Intrastate Air... Air Quality Control Regions § 81.158 Southern Wisconsin Intrastate Air Quality Control Region. The Southern Wisconsin Intrastate Air Quality Control Region consists of the territorial area encompassed by...

40 CFR 81.158 - Southern Wisconsin Intrastate Air Quality Control Region.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 18 2012-07-01 2012-07-01 false Southern Wisconsin Intrastate Air... Air Quality Control Regions § 81.158 Southern Wisconsin Intrastate Air Quality Control Region. The Southern Wisconsin Intrastate Air Quality Control Region consists of the territorial area encompassed by...

The Southern Cone: A critical element in North American geology

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

Dalziel, I.W.D.

1993-02-01

The Pacific and Atlantic-Gulf of Mexico continental margins converge towards southern Mexico, delimiting the Southern Cone of North American. The margins are controlled by late Precambrian to early Paleozoic rift systems. The Neoproterozoic rifts along the Pacific margin truncate the 1.3--1.0 Ga Grenville-Llano front and still older structural boundaries within the craton, such as the Snowbird line. The Atlantic margin originated by separation from another continent within the Grenville orogen near the time of the Precambrian-Cambrian boundary. The Gulf of Mexico margin was initiated with rifting at that time, but appears to truncate the Ordovician Taconian orogen in Georgia. Themore » continental margins of the Southern Cone may prove critical in understanding the origin of North America as a discrete continent. A possible continuation of the Grenville-Llano front has now been identified along the Pacific margin of the East Antarctic craton; the opposite side of the Grenville orogen may be present in South America and East Antarctic; a southern continuation of the Taconic Appalachians may have been identified in southern South American and Antarctica (L. Dalla Salda et al., Geology, 1992 a;b: I. Dalziel, Geology, 1991, and GSA Today, 1992; P. Hoffman, Science, 1991; E. Moores, Geology, 1991). Thus the geology of the Southern Cone of North America provides opportunities for critical testing of these globally important hypotheses, notably through geochronometry, isotope geochemistry, stratigraphy, and paleobiogeography. Conversely, East Antarctica, southern Africa, and the proto-Andean margin of South America may offer exciting opportunities to further understanding of pre-Pangea geology across southern North America.« less

Spatio-temporal patterns of Ulva prolifera blooms and the corresponding influence on chlorophyll-a concentration in the Southern Yellow Sea, China.

PubMed

Sun, Xiao; Wu, Mengquan; Xing, Qianguo; Song, Xiaodong; Zhao, Deheng; Han, Qianqian; Zhang, Guangzong

2018-06-04

The world's largest macroalgal blooms (MABs) caused by the Ulva prolifera outbreaks have occurred every summer since 2007 in the Southern Yellow Sea, China. Accumulating evidence showed that MABs may deteriorate the regional marine environment and influence the growth of some primary producers such as phytoplankton. In this study, we investigated the spatio-temporal patterns of U. prolifera green tides and chlorophyll-a concentration in the Southern Yellow Sea in 2015 using satellite images obtained from HJ-1 CCD, MODIS, and GOCI. The correlation between the distributions of U. prolifera abundance and chlorophyll-a concentration was analyzed quantitatively by setting up a series of 5 × 5 km experimental grids, and we also discussed the possible mechanisms about the influence of U. prolifera blooms on the other floating microalgae. The results showed that the development of U. prolifera blooms in the Southern Yellow Sea in 2015 could be featured as "appearance - development - outbreak - decline - disappearance", while the concentration of chlorophyll-a showed "increase - sharp decline - slow recovery - stabilization" from April to August. We also found that the concentration of chlorophyll-a had the following relationships with U. prolifera temporally: (1) the concentration of chlorophyll-a increased with the growth of U. prolifera from April to mid-May; (2) the chlorophyll-a concentration decreased sharply with the dramatically increased coverage of U. prolifera in June; and (3) the chlorophyll-a concentration slowly recovered and finally stabilized as U. prolifera decreased in July. Generally, there was a negative correlation between the occurrence of U. prolifera and chlorophyll-a concentration in the Southern Yellow Sea, China. Our results showed that the outbreak of U. prolifera does have a certain impact on the growth and reproduction of planktonic microalgae, and it suggests that U. prolifera blooms have potentially altered the ecological balance in

The Southern Manifesto as Education Policy in Mississippi

ERIC Educational Resources Information Center

Luckett, Robert, Jr.

2016-01-01

In 1956, southern Congressmen signed the Southern Manifesto, rejecting the Supreme Court's "Brown v. Board of Education" ruling. This moment, in the general American consciousness, marked the rise of White massive resistance to Black advancement, a racist foray doomed to be swept aside by civil rights forces and a determined federal…

Southern regional report

Treesearch

Bryce J Stokes

1997-01-01

It appears as if many in wood procurement, forest management, operations, manufacturing, and sales in the Southern U.S. are simultaneously biting the bullet and showing signs of guarded optimism for the future. On the one hand, during last year, purchasing costs were high, selling prices were low, quotas ruled, and machine purchases sagged. However, there is continued...

Stable chloroplast transformation of immature scutella and inflorescences in wheat (Triticum aestivum L.).

PubMed

Cui, Cuiju; Song, Fei; Tan, Yi; Zhou, Xuan; Zhao, Wen; Ma, Fengyun; Liu, Yunyi; Hussain, Javeed; Wang, Yuesheng; Yang, Guangxiao; He, Guangyuan

2011-04-01

Chloroplast transformation in wheat was achieved by bombardment of scutella from immature embryos and immature inflorescences, respectively. A wheat chloroplast site-specific expression vector, pBAGNRK, was constructed by placing an expression cassette containing neomycin phosphotransferase II (nptII) and green fluorescent protein (gfp) as selection and reporter genes, respectively, in the intergenic spacer between atpB and rbcL of wheat chloroplast genome. Integration of gfp gene in the plastome was identified by polymerase chain reaction (PCR) analysis and Southern blotting using gfp gene as a probe. Expression of GFP protein was examined by western blot. Three positive transformants were obtained and the Southern blot of partial fragment of atpB and rbcL (targeting site) probes verified that one of them was homoplasmic. Stable expression of GFP fluorescence was confirmed by confocal microscopy in the leaf tissues from T(1) progeny seedlings. PCR analysis of gfp gene also confirmed the inheritance of transgene in the T(1) progeny. These results strengthen the feasibility of wheat chloroplast transformation and also give a novel method for the introduction of important agronomic traits in wheat through chloroplast transformation.

Characterization of cDNA encoding molt-inhibiting hormone of the crab, Cancer pagurus; expression of MIH in non-X-organ tissues.

PubMed

Lu, W; Wainwright, G; Olohan, L A; Webster, S G; Rees, H H; Turner, P C

2001-10-31

Synthesis of ecdysteroids (molting hormones) by crustacean Y-organs is regulated by a neuropeptide, molt-inhibiting hormone (MIH), produced in eyestalk neural ganglia. We report here the molecular cloning of a cDNA encoding MIH of the edible crab, Cancer pagurus. Full-length MIH cDNA was obtained by using reverse transcription-polymerase chain reaction (RT-PCR) with degenerate oligonucleotides based upon the amino acid sequence of MIH, in conjunction with 5'- and 3'-RACE. Full-length clones of MIH cDNA were obtained that encoded a 35 amino acid putative signal peptide and the mature 78 amino acid peptide. Of various tissues examined by Northern blot analysis, the X-organ was the sole major site of expression of the MIH gene. However, a nested-PCR approach using non-degenerate MIH-specific primers indicated the presence of MIH transcripts in other tissues. Southern blot analysis indicated a simple gene arrangement with at least two copies of the MIH gene in the genome of C. pagurus. Additional Southern blotting experiments detected MIH-hybridizing bands in another Cancer species, Cancer antennarius and another crab species, Carcinus maenas.

Detection of the HTLV-I gene on cytologic smear slides.

PubMed

Kashima, Kenji; Nagahama, Junji; Sato, Keiji; Tanamachi, Hiroyuki; Gamachi, Ayako; Daa, Tsutomu; Nakayama, Iwao; Yokoyama, Shigeo

2002-01-01

To apply the polymerase chain reaction (PCR) for detection of the HTLV-I gene from cytologic smear slides. Samples were from seven cases of serum anti-ATL antibody (ATLA)-positive T-cell lymphoma and three from ATLA-negative T-cell lymphoma. Six of the seven ATLA-positive cases were confirmed to be ATLL by Southern blotting. From the seventh case a fresh sample for blotting could not obtained. DNA was extracted from the cytologic smear slides of all 10 cases; they had been stained with Papanicolaou or May-Giemsa stain, digested with proteinase K and precipitated with phenol and ethanol. The target sequence in the pX region of the HTLV-I gene was amplified by PCR. All seven ATLA-positive cases, including one that had not yet been confirmed by Southern blotting, showed a single band, as predicted, while the three ATLA-negative cases showed no band. If cytologic smear slides are available but a fresh sample is not, the PCR method should provide evidence that the virus is present since in our study sufficient DNA templates were successfully extracted from the stained cytologic smear slides for detection of the virus.

The Angola Low: relationship with southern African rainfall and ENSO

NASA Astrophysics Data System (ADS)

Crétat, Julien; Pohl, Benjamin; Dieppois, Bastien; Berthou, Ségolène; Pergaud, Julien

2018-05-01

The main states of the Angola Low (AL) are identified using clustering analysis applied to daily anomalous patterns of 700-hPa wind vorticity over Angola and adjacent countries from November to March for the 1980/81-2014/15 period. At the daily timescale, we examine the extent to which the main states of the AL modulate daily rainfall over southern Africa. At the interannual timescale, we assess both the relationship between the occurrence of these AL states and El Niño southern oscillation (ENSO) and the role of the AL in explaining ENSO's failure in driving southern African rainfall at times. Three reanalyses are considered to account for uncertainties induced by the scarcity of data available for assimilation over southern Africa. Three preferential states of the Angola Low are identified: AL state close to its seasonal climatology with slight zonal displacements, anomalously weak AL state and anomalously strong AL state with meridional displacements. These different states all significantly modulate daily southern African rainfall. Near-climatological AL state promotes wet rainfall anomalies over eastern subtropical southern Africa and dry rainfall anomalies over its western part. A slight westward shift in the near-climatological position of the AL leads to reversed zonal gradient in rainfall. The remaining regimes significantly modulate the meridional gradient in southern African rainfall. Anomalously weak and anomalously northward AL states promote wet rainfall anomalies over tropical southern Africa and dry rainfall anomalies over subtropical southern Africa. The reverse prevails for anomalously southward AL. At the interannual timescale, ENSO significantly modulates the seasonal occurrence of most AL states in the three reanalyses. Anomalously weak and southward AL states are more strongly correlated with regional rainfall than ENSO in all reanalyses, suggesting that accounting for AL variability may improve seasonal forecasts. Case study analysis of the

Book review: Southern Forested Wetlands: Ecology and Management

Treesearch

Carl C. Trettin

2000-01-01

The southern region has the largest proportion of wetlands in the conterminous US. The majority of that wetland resource is forested by diverse vegetation communities reflecting differences in soil, hydrology, geomorphology, climatic conditions and past management. Wetland resources in the southern US are very important to the economy providing both commodity and non-...

Industrial Physics---Southern California Style

NASA Astrophysics Data System (ADS)

Leslie, Stuart

2013-03-01

Only in Southern California did space-age style really come into its own as a unique expression of Cold War scientific culture. The corporate campuses of General Atomic in San Diego and North American Aviation in Los Angeles perfectly expressed the exhilarating spirit of Southern California's aerospace era, scaling up the residential version of California modernism to industrial proportion. Architects William Pereira and A.C. Martin Jr., in collaboration with their scientific counterparts, fashioned military-industrial `dream factories' for industrial physics that embodied the secret side of the space-age zeitgeist, one the public could only glimpse of in photographs, advertisements, and carefully staged open houses. These laboratories served up archetypes of the California dream for a select audience of scientists, engineers, and military officers, live-action commercials for a lifestyle intended to lure the best and brightest to Southern California. Paradoxically, they hid in plain sight, in the midst of aerospace suburbs, an open secret, at once visible and opaque, the public face of an otherwise invisible empire. Now, at the end of the aerospace era, these places have become an endangered species, difficult to repurpose, on valuable if sometimes highly polluted land. Yet they offer an important reminder of a more confident time when many physicists set their sights on the stars.

Simultaneous Detection of Bovine Theileria and Babesia Species by Reverse Line Blot Hybridization

PubMed Central

Gubbels, J. M.; de Vos, A. P.; van der Weide, M.; Viseras, J.; Schouls, L. M.; de Vries, E.; Jongejan, F.

1999-01-01

A reverse line blot (RLB) assay was developed for the identification of cattle carrying different species of Theileria and Babesia simultaneously. We included Theileria annulata, T. parva, T. mutans, T. taurotragi, and T. velifera in the assay, as well as parasites belonging to the T. sergenti-T. buffeli-T. orientalis group. The Babesia species included were Babesia bovis, B. bigemina, and B. divergens. The assay employs one set of primers for specific amplification of the rRNA gene V4 hypervariable regions of all Theileria and Babesia species. PCR products obtained from blood samples were hybridized to a membrane onto which nine species-specific oligonucleotides were covalently linked. Cross-reactions were not observed between any of the tested species. No DNA sequences from Bos taurus or other hemoparasites (Trypanosoma species, Cowdria ruminantium, Anaplasma marginale, and Ehrlichia species) were amplified. The sensitivity of the assay was determined at 0.000001% parasitemia, enabling detection of the carrier state of most parasites. Mixed DNAs from five different parasites were correctly identified. Moreover, blood samples from cattle experimentally infected with two different parasites reacted only with the corresponding species-specific oligonucleotides. Finally, RLB was used to screen blood samples collected from carrier cattle in two regions of Spain. T. annulata, T. orientalis, and B. bigemina were identified in these samples. In conclusion, the RLB is a versatile technique for simultaneous detection of all bovine tick-borne protozoan parasites. We recommend its use for integrated epidemiological monitoring of tick-borne disease, since RLB can also be used for screening ticks and can easily be expanded to include additional hemoparasite species. PMID:10325324

Epidemiology of Rheumatoid Arthritis in Southern Albania.

PubMed

Koko, Vjollca; Ndrepepa, Ana; Skenderaj, Skender

2015-06-01

The aim of this study was to assess the prevalence, incidence and the burden of rheumatoid arthritis (RA) in the Southern Albania. This is an epidemiologic observational study with cross-sectional analyses of all patients with RA who lived in Southern Albania during the 1995-2011 years. The RA prevalence, incidence and disability-adjusted life years (DALY) were assessed. During the 1995-2011 years, 194 patients (154 females and 40 males) with RA living in Southern Albania were identified. The prevalence of RA in 2011 was 0.25% in general population and 0.34% in adult (>14 years) population. The incidence of RA in 2011 was 0.012% (12 new cases per 100000 inhabitants) and 0.016% (16 new cases per 100000 adults). The prevalence increased (from 0.036% in 1996 to 0.25% in 2011) and the incidence did not change over the study period. The mortality was 3.2% (n=7 deaths). The DALY due to RA was 823 years per 100000 inhabitants during 1995-2011 years. RA in Southern Albania has a prevalence of 0.25 % and an annual incidence of 0.012% in the general population in 2011. RA was responsible for a considerable burden on the health of population during the 1995-2011 years.

Trends in Southern Ocean Eddy Kinetic Energy

NASA Astrophysics Data System (ADS)

Chambers, Don

2016-04-01

A recent study by Hogg et al. (JGR, 2015) has demonstrated a 20-year trend in eddy kinetic energy (EKE) computed from satellite altimetry data. However, this estimate is based on an averaging over large spatial areas. In this study, we use the same methods to examine regional EKE trends throughout the Southern Ocean, from 1993-2015. We do find significant positive trends in several areas of the Southern Ocean, mainly in regions with high mean EKE associated with interactions between jets and bathymetry. At the same time, however, there are also regions with significant negative trends. Overall, EKE in the majority of the Southern Ocean has not changed. These results suggest that the estimates of Hogg et al. may have been biased by these regional extremes, and that more work is needed to quantify climatic changes in EKE.

Trends in Southern Ocean Eddy Kinetic Energy

NASA Astrophysics Data System (ADS)

Chambers, D. P.

2016-02-01

A recent study by Hogg et al. (JGR, 2015) has demonstrated a 20-year trend in eddy kinetic energy (EKE) computed from satellite altimetry data. However, this estimate is based on an averaging over large spatial areas. In this study, we use the same methods to examine regional EKE trends throughout the Southern Ocean, from 1993-2015. We do find significant positive trends in several areas of the Southern Ocean, mainly in regions with high mean EKE associated with interactions between jets and bathymetry. At the same time, however, there are also regions with significant negative trends. Overall, EKE in the majority of the Southern Ocean has not changed. These results suggest that the estimates of Hogg et al. may have been biased by these regional extremes, and that more work is needed to quantify climatic changes in EKE.

Atmospheric Chemistry Over Southern Africa

NASA Technical Reports Server (NTRS)

Gatebe, Charles K.; Levy, Robert C.; Thompson, Anne M.

2011-01-01

During the southern African dry season, regional haze from mixed industrial pollution, biomass burning aerosol and gases from domestic and grassland fires, and biogenic sources from plants and soils is worsened by a semi-permanent atmosphere gyre over the subcontinent. These factors were a driver of several major international field campaigns in the 1990s and early 2000s, and attracted many scientists to the region. Some researchers were interested in understanding fundamental processes governing chemistry of the atmosphere and interaction with climate change. Others found favorable conditions for evaluating satellite-derived measurements of atmospheric properties and a changing land surface. With that background in mind a workshop on atmospheric chemistry was held in South Africa. Sponsored by the International Commission for Atmospheric Chemistry and Global Pollution (ICACGP; http://www.icacgp.org/), the workshop received generous support from the South African power utility, Eskom, and the Climatology Research Group of the University of the Witwatersrand, Johannesburg, South Africa. The purpose of the workshop was to review some earlier findings as well as more recent findings on southern African climate vulnerability, chemical changes due to urbanization, land-use modification, and how these factors interact. Originally proposed by John Burrows, president of ICACGP, the workshop was the first ICACGP regional workshop to study the interaction of air pollution with global chemical and climate change. Organized locally by the University of the Witwatersrand, the workshop attracted more than 60 delegates from South Africa, Mozambique, Botswana, Zimbabwe, France, Germany, Canada, and the United States. More than 30 presentations were given, exploring both retrospective and prospective aspects of the science. In several talks, attention was focused on southern African chemistry, atmospheric pollution monitoring, and climate processes as they were studied in the field

The earliest unequivocally modern humans in southern China.

PubMed

Liu, Wu; Martinón-Torres, María; Cai, Yan-jun; Xing, Song; Tong, Hao-wen; Pei, Shu-wen; Sier, Mark Jan; Wu, Xiao-hong; Edwards, R Lawrence; Cheng, Hai; Li, Yi-yuan; Yang, Xiong-xin; de Castro, José María Bermúdez; Wu, Xiu-jie

2015-10-29

The hominin record from southern Asia for the early Late Pleistocene epoch is scarce. Well-dated and well-preserved fossils older than ∼45,000 years that can be unequivocally attributed to Homo sapiens are lacking. Here we present evidence from the newly excavated Fuyan Cave in Daoxian (southern China). This site has provided 47 human teeth dated to more than 80,000 years old, and with an inferred maximum age of 120,000 years. The morphological and metric assessment of this sample supports its unequivocal assignment to H. sapiens. The Daoxian sample is more derived than any other anatomically modern humans, resembling middle-to-late Late Pleistocene specimens and even contemporary humans. Our study shows that fully modern morphologies were present in southern China 30,000-70,000 years earlier than in the Levant and Europe. Our data fill a chronological and geographical gap that is relevant for understanding when H. sapiens first appeared in southern Asia. The Daoxian teeth also support the hypothesis that during the same period, southern China was inhabited by more derived populations than central and northern China. This evidence is important for the study of dispersal routes of modern humans. Finally, our results are relevant to exploring the reasons for the relatively late entry of H. sapiens into Europe. Some studies have investigated how the competition with H. sapiens may have caused Neanderthals' extinction (see ref. 8 and references therein). Notably, although fully modern humans were already present in southern China at least as early as ∼80,000 years ago, there is no evidence that they entered Europe before ∼45,000 years ago. This could indicate that H. neanderthalensis was indeed an additional ecological barrier for modern humans, who could only enter Europe when the demise of Neanderthals had already started.

North-Seeking Magnetotactic Gammaproteobacteria in the Southern Hemisphere

PubMed Central

Leão, Pedro; Teixeira, Lia C. R. S.; Cypriano, Jefferson; Farina, Marcos; Abreu, Fernanda; Bazylinski, Dennis A.

2016-01-01

ABSTRACT Magnetotactic bacteria (MTB) comprise a phylogenetically diverse group of prokaryotes capable of orienting and navigating along magnetic field lines. Under oxic conditions, MTB in natural environments in the Northern Hemisphere generally display north-seeking (NS) polarity, swimming parallel to the Earth's magnetic field lines, while those in the Southern Hemisphere generally swim antiparallel to magnetic field lines (south-seeking [SS] polarity). Here, we report a population of an uncultured, monotrichously flagellated, and vibrioid MTB collected from a brackish lagoon in Brazil in the Southern Hemisphere that consistently exhibits NS polarity. Cells of this organism were mainly located below the oxic-anoxic interface (OAI), suggesting it is capable of some type of anaerobic metabolism. Magnetosome crystalline habit and composition were consistent with elongated prismatic magnetite (Fe3O4) particles. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that this organism belongs to a distinct clade of the Gammaproteobacteria class. The presence of NS MTB in the Southern Hemisphere and the previously reported finding of SS MTB in the Northern Hemisphere reinforce the idea that magnetotaxis is more complex than we currently understand and may be modulated by factors other than O2 concentration and redox gradients in sediments and water columns. IMPORTANCE Magnetotaxis is a navigational mechanism used by magnetotactic bacteria to move along geomagnetic field lines and find an optimal position in chemically stratified sediments. For that, magnetotactic bacteria swim parallel to the geomagnetic field lines under oxic conditions in the Northern Hemisphere, whereas those in the Southern Hemisphere swim antiparallel to magnetic field lines. A population of uncultured vibrioid magnetotactic bacteria was discovered in a brackish lagoon in the Southern Hemisphere that consistently swim northward, i.e., the opposite of the overwhelming majority of other

Large-Scale Weather Disturbances in Mars’ Southern Extratropics

NASA Astrophysics Data System (ADS)

Hollingsworth, Jeffery L.; Kahre, Melinda A.

2015-11-01

Between late autumn and early spring, Mars’ middle and high latitudes within its atmosphere support strong mean thermal gradients between the tropics and poles. Observations from both the Mars Global Surveyor (MGS) and Mars Reconnaissance Orbiter (MRO) indicate that this strong baroclinicity supports intense, large-scale eastward traveling weather systems (i.e., transient synoptic-period waves). These extratropical weather disturbances are key components of the global circulation. Such wave-like disturbances act as agents in the transport of heat and momentum, and generalized scalar/tracer quantities (e.g., atmospheric dust, water-vapor and ice clouds). The character of large-scale, traveling extratropical synoptic-period disturbances in Mars' southern hemisphere during late winter through early spring is investigated using a moderately high-resolution Mars global climate model (Mars GCM). This Mars GCM imposes interactively lifted and radiatively active dust based on a threshold value of the surface stress. The model exhibits a reasonable "dust cycle" (i.e., globally averaged, a dustier atmosphere during southern spring and summer occurs). Compared to their northern-hemisphere counterparts, southern synoptic-period weather disturbances and accompanying frontal waves have smaller meridional and zonal scales, and are far less intense. Influences of the zonally asymmetric (i.e., east-west varying) topography on southern large-scale weather are examined. Simulations that adapt Mars’ full topography compared to simulations that utilize synthetic topographies emulating key large-scale features of the southern middle latitudes indicate that Mars’ transient barotropic/baroclinic eddies are highly influenced by the great impact basins of this hemisphere (e.g., Argyre and Hellas). The occurrence of a southern storm zone in late winter and early spring appears to be anchored to the western hemisphere via orographic influences from the Tharsis highlands, and the Argyre

Qualitative, semi-quantitative, and quantitative simulation of the osmoregulation system in yeast

PubMed Central

Pang, Wei; Coghill, George M.

2015-01-01

In this paper we demonstrate how Morven, a computational framework which can perform qualitative, semi-quantitative, and quantitative simulation of dynamical systems using the same model formalism, is applied to study the osmotic stress response pathway in yeast. First the Morven framework itself is briefly introduced in terms of the model formalism employed and output format. We then built a qualitative model for the biophysical process of the osmoregulation in yeast, and a global qualitative-level picture was obtained through qualitative simulation of this model. Furthermore, we constructed a Morven model based on existing quantitative model of the osmoregulation system. This model was then simulated qualitatively, semi-quantitatively, and quantitatively. The obtained simulation results are presented with an analysis. Finally the future development of the Morven framework for modelling the dynamic biological systems is discussed. PMID:25864377

Rigour in quantitative research.

PubMed

Claydon, Leica Sarah

2015-07-22

This article which forms part of the research series addresses scientific rigour in quantitative research. It explores the basis and use of quantitative research and the nature of scientific rigour. It examines how the reader may determine whether quantitative research results are accurate, the questions that should be asked to determine accuracy and the checklists that may be used in this process. Quantitative research has advantages in nursing, since it can provide numerical data to help answer questions encountered in everyday practice.

Southern Pine Seed Sources

Treesearch

Ronald C. Schmidtling

2001-01-01

The selection of an appropriate seed source is critical for successful southern pine plantations. Guidelines for selection of seed sources are presented for loblolly (Pinus taeda L.), slash (P. elliottii Engelm.), longleaf (P. palustris Mill.), Virginia (P. virginiana Mill.), shortleaf (P. echinata...

Dioscin Inhibits HSC-T6 Cell Migration via Adjusting SDC-4 Expression: Insights from iTRAQ-Based Quantitative Proteomics.

PubMed

Yin, Lianhong; Qi, Yan; Xu, Youwei; Xu, Lina; Han, Xu; Tao, Xufeng; Song, Shasha; Peng, Jinyong

2017-01-01

Hepatic stellate cells (HSCs) migration, an important bioprocess, contributes to the development of liver fibrosis. Our previous studies have found the potent activity of dioscin against liver fibrosis by inhibiting HSCs proliferation, triggering the senescence and inducing apoptosis of activated HSCs, but the molecular mechanisms associated with cell migration were not clarified. In this work, iTRAQ (isobaric tags for relative and absolution quantitation)-based quantitative proteomics study was carried out, and a total of 1566 differentially expressed proteins with fold change ≥2.0 and p < 0.05 were identified in HSC-T6 cells treated by dioscin (5.0 μg/mL). Based on Gene Ontology classification, String and KEGG pathway assays, the effects of dioscin to inhibit cell migration via regulating SDC-4 were carried out. The results of wound-healing, cell migration and western blotting assays indicated that dioscin significantly inhibit HSC-T6 cell migration through SDC-4-dependent signal pathway by affecting the expression levels of Fn, PKCα, Src, FAK, and ERK1/2. Specific SDC-4 knockdown by shRNA also blocked HSC-T6 cell migration, and dioscin slightly enhanced the inhibiting effect. Taken together, the present work showed that SDC-4 played a crucial role on HSC-T6 cell adhesion and migration of dioscin against liver fibrosis, which may be one potent therapeutic target for fibrotic diseases.

Silviculture and management strategies applicable to southern hardwoods

Treesearch

Ray R. Hicks; William H. Conner; Robert C. Kellison; David Van Lear

2004-01-01

Southern hardwood forests stretch from the Virginias to Florida and from the mid-Atlantic to Missouri. They can generally be grouped into upland forests and bottomland forests. The upland hardwood forests of the southern region are usually associated with the mountainous topography of the Appalachians and Ozarks. Bottomland hardwoods are found along the floodplains of...

A new tree classification system for southern hardwoods

Treesearch

James S. Meadows; Daniel A. Jr. Skojac

2008-01-01

A new tree classification system for southern hardwoods is described. The new system is based on the Putnam tree classification system, originally developed by Putnam et al., 1960, Management ond inventory of southern hardwoods, Agriculture Handbook 181, US For. Sew., Washington, DC, which consists of four tree classes: (1) preferred growing stock, (2) reserve growing...

Southern California Earthquake Center Geologic Vertical Motion Database

NASA Astrophysics Data System (ADS)

Niemi, Nathan A.; Oskin, Michael; Rockwell, Thomas K.

2008-07-01

The Southern California Earthquake Center Geologic Vertical Motion Database (VMDB) integrates disparate sources of geologic uplift and subsidence data at 104- to 106-year time scales into a single resource for investigations of crustal deformation in southern California. Over 1800 vertical deformation rate data points in southern California and northern Baja California populate the database. Four mature data sets are now represented: marine terraces, incised river terraces, thermochronologic ages, and stratigraphic surfaces. An innovative architecture and interface of the VMDB exposes distinct data sets and reference frames, permitting user exploration of this complex data set and allowing user control over the assumptions applied to convert geologic and geochronologic information into absolute uplift rates. Online exploration and download tools are available through all common web browsers, allowing the distribution of vertical motion results as HTML tables, tab-delimited GIS-compatible text files, or via a map interface through the Google Maps™ web service. The VMDB represents a mature product for research of fault activity and elastic deformation of southern California.

Climate and southern Africa's water-energy-food nexus

NASA Astrophysics Data System (ADS)

Conway, Declan; van Garderen, Emma Archer; Deryng, Delphine; Dorling, Steve; Krueger, Tobias; Landman, Willem; Lankford, Bruce; Lebek, Karen; Osborn, Tim; Ringler, Claudia; Thurlow, James; Zhu, Tingju; Dalin, Carole

2015-09-01

In southern Africa, the connections between climate and the water-energy-food nexus are strong. Physical and socioeconomic exposure to climate is high in many areas and in crucial economic sectors. Spatial interdependence is also high, driven, for example, by the regional extent of many climate anomalies and river basins and aquifers that span national boundaries. There is now strong evidence of the effects of individual climate anomalies, but associations between national rainfall and gross domestic product and crop production remain relatively weak. The majority of climate models project decreases in annual precipitation for southern Africa, typically by as much as 20% by the 2080s. Impact models suggest these changes would propagate into reduced water availability and crop yields. Recognition of spatial and sectoral interdependencies should inform policies, institutions and investments for enhancing water, energy and food security. Three key political and economic instruments could be strengthened for this purpose: the Southern African Development Community, the Southern African Power Pool and trade of agricultural products amounting to significant transfers of embedded water.

Natural History of the Southern Pine Beetle

Treesearch

Fred P. Hain; Adrian J. Duehl; Micah J. Gardner; Thomas L. Payne

2011-01-01

The southern pine beetle (SPB) is a tree killer of southern yellow pines. All life stages—eggs, larvae, pupae, and adults—infest the inner bark or phloem tissue of the host tree. Adult beetles overcome the tree’s defenses through a mass-attack phenomenon. They are attracted to the tree by a pheromone system consisting of volatiles produced by the beetles and the host....

New Method for Simultaneous Species-Specific Identification of Equine Strongyles (Nematoda, Strongylida) by Reverse Line Blot Hybridization▿

PubMed Central

Traversa, Donato; Iorio, Raffaella; Klei, Thomas R.; Kharchenko, Vitaliy A.; Gawor, Jakub; Otranto, Domenico; Sparagano, Olivier A. E.

2007-01-01

The ability of a reverse line blot (RLB) assay to identify 13 common species of equine small strongyles (cyathostomins) and to discriminate them from three Strongylus spp. (large strongyles) was demonstrated. The assay relied on the specific hybridization of PCR-amplified intergenic spacer DNA fragments of the nuclear ribosomal DNA to membrane-bound species-specific probes. All cyathostomins examined were unequivocally identified and simultaneously discriminated from each other and from three large strongyles (Strongylus edentatus, Strongylus equinus, and Strongylus vulgaris). This assay will enable the accurate and rapid identification of equine cyathostomins irrespective of their life cycle stage, opening important avenues for a better understanding of their biology and epidemiology and of the pathogenesis of cyathostomin-associated disease. In particular, this RLB method promises to be a powerful diagnostic tool to determine the roles of individual species in the pathogenesis of mixed infections and to elucidate some aspects of cyathostominosis. Also, it could represent a basic step toward the development of a rapid and simple molecular test for the early detection of drug-resistant genotypes of horse strongyle species. PMID:17626168

Practical Molecular Biology for Students: An Integrated Approach to Teaching Basic Techniques.

ERIC Educational Resources Information Center

Hames, B. David; And Others

1990-01-01

An activity that introduces students to the correct handling of bacterial recombinants, antibiotic sensitivity testing, insertional inactivation, plasmid DNA isolation, restriction endonuclease digestion, agarose gel electrophoresis, Southern blotting, hybridization, and autoradiography is presented. A list of needed materials, procedures, safety…

The forest-land owners of southern New England

Treesearch

Neal P. Kingsley

1976-01-01

A statistical-analytical report of a mail canvass of the owners of privately owned commercial forest land in the three Southern New England States-Connecticut, Massachusetts, and Rhode Island. The study was conducted in conjunction with the second forest survey of Southern New England by the USDA Forest Service. Trends in forest-land ownership and the attitudes and...

New cooperative seismograph networks established in southern California

USGS Publications Warehouse

Hill, D.P.

1974-01-01

Southern California has more active faults located close to large, urban population centers than any other region in the United States. Reduction of risk to life and property posed by potential earthquakes along these active faults is a primary motivation for a cooperative earthquake research program between the U.S Geological Survey and major universities in Southern California. 

Critical Quantitative Inquiry in Context

ERIC Educational Resources Information Center

Stage, Frances K.; Wells, Ryan S.

2014-01-01

This chapter briefly traces the development of the concept of critical quantitative inquiry, provides an expanded conceptualization of the tasks of critical quantitative research, offers theoretical explanation and justification for critical research using quantitative methods, and previews the work of quantitative criticalists presented in this…

Improved and high throughput quantitative measurements of weak GFP expression in transgenic plant materials.

PubMed

Wu, Jing-Jing; Liu, Yu-Wen; Sun, Meng-Xiang

2011-07-01

Green fluorescent proteins (GFPs) are widely used in tracing transgene expression and have been known as convenient and efficient markers for plant transformation. However, sometimes researchers are still puzzled by the weak fluorescence since it makes the observation of GFP signals and confirmation of transgenic plants difficult. In this investigation, we explored the possibility of enhancing the weak signals by changing the pH environment of detection and took microplate reader as a more effective instrument compared to traditional fluorescent microscope to detect the weak signals. It was found that the fluorescence intensity of enhanced GFP (EGFP) in transgenic plants can be increased 2-6 folds by altering the environmental pH, and the concentration of EGFP at a large scale (ranged from 20 ng/ml to 20 μg/ml) can be detected and quantified. It can exclude the influence of degradation fragment and hence facilitate later analysis; these advantages were further verified by comparing with western blotting and confocal microscopy. It was reliable and effective for the qualitative and quantitative analysis of transgenic plants and was more suitable for the detection of very weak fluorescent signals.

Quantitative interpretation of atmospheric carbon records over the last glacial termination

NASA Astrophysics Data System (ADS)

KöHler, Peter; Fischer, Hubertus; Munhoven, Guy; Zeebe, Richard E.

2005-12-01

The glacial/interglacial rise in atmospheric pCO2 is one of the best known changes in paleoclimate research, yet the cause for it is still unknown. Forcing the coupled ocean-atmosphere-biosphere box model of the global carbon cycle BICYCLE with proxy data over the last glacial termination, we are able to quantitatively reproduce transient variations in pCO2 and its isotopic signatures (δ13C, Δ14C) observed in natural climate archives. The sensitivity of the Box model of the Isotopic Carbon cYCLE (BICYCLE) to high or low latitudinal changes is comparable to other multibox models or more complex ocean carbon cycle models, respectively. The processes considered here ranked by their contribution to the glacial/interglacial rise in pCO2 in decreasing order are: the rise in Southern Ocean vertical mixing rates (>30 ppmv), decreases in alkalinity and carbon inventories (>30 ppmv), the reduction of the biological pump (˜20 ppmv), the rise in ocean temperatures (15-20 ppmv), the resumption of ocean circulation (15-20 ppmv), and coral reef growth (<5 ppmv). The regrowth of the terrestrial biosphere, sea level rise and the increase in gas exchange through reduced sea ice cover operate in the opposite direction, decreasing pCO2 during Termination I by ˜30 ppmv. According to our model the sequence of events during Termination I might have been the following: a reduction of aeolian iron fertilization in the Southern Ocean together with a breakdown in Southern Ocean stratification, the latter caused by rapid sea ice retreat, trigger the onset of the pCO2 increase. After these events the reduced North Atlantic Deep Water (NADW) formation during the Heinrich 1 event and the subsequent resumption of ocean circulation at the beginning of the Bølling-Allerød warm interval are the main processes determining the atmospheric carbon records in the subsequent time period of Termination I. We further deduce that a complete shutdown of the NADW formation during the Younger Dryas was

Southern Hemisphere Neutron Map

NASA Image and Video Library

2002-03-01

NASA Mars Odyssey spacecraft produced this high-energy neutron detector map of neutrons in Mars southern hemisphere. The blue region around the south pole indicates a high content of hydrogen in the upper 2 to 3 meters 7 to 10 feet of the surface.

Qualitative, semi-quantitative, and quantitative simulation of the osmoregulation system in yeast.

PubMed

Pang, Wei; Coghill, George M

2015-05-01

In this paper we demonstrate how Morven, a computational framework which can perform qualitative, semi-quantitative, and quantitative simulation of dynamical systems using the same model formalism, is applied to study the osmotic stress response pathway in yeast. First the Morven framework itself is briefly introduced in terms of the model formalism employed and output format. We then built a qualitative model for the biophysical process of the osmoregulation in yeast, and a global qualitative-level picture was obtained through qualitative simulation of this model. Furthermore, we constructed a Morven model based on existing quantitative model of the osmoregulation system. This model was then simulated qualitatively, semi-quantitatively, and quantitatively. The obtained simulation results are presented with an analysis. Finally the future development of the Morven framework for modelling the dynamic biological systems is discussed. Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.

Quantitative dispersion microscopy

PubMed Central

Fu, Dan; Choi, Wonshik; Sung, Yongjin; Yaqoob, Zahid; Dasari, Ramachandra R.; Feld, Michael

2010-01-01

Refractive index dispersion is an intrinsic optical property and a useful source of contrast in biological imaging studies. In this report, we present the first dispersion phase imaging of living eukaryotic cells. We have developed quantitative dispersion microscopy based on the principle of quantitative phase microscopy. The dual-wavelength quantitative phase microscope makes phase measurements at 310 nm and 400 nm wavelengths to quantify dispersion (refractive index increment ratio) of live cells. The measured dispersion of living HeLa cells is found to be around 1.088, which agrees well with that measured directly for protein solutions using total internal reflection. This technique, together with the dry mass and morphology measurements provided by quantitative phase microscopy, could prove to be a useful tool for distinguishing different types of biomaterials and studying spatial inhomogeneities of biological samples. PMID:21113234

[Vaccines: producers in countries of the Southern hemisphere].

PubMed

Bertrand, J J

2007-08-01

Vaccine producers in southern hemisphere countries now contribute significantly to global output. In 2006 southern hemisphere countries accounted for more than 10% of the total worldwide production with a progression approximately 70% greater than all producers combined in the two-year period between 2004 and 2006. Though difficult to measure, production in volume is higher due to lower prices practiced in most of these countries. For many years before the 1980s, production was scattered among numerous limited-scale companies. Most were founded at the initiative of governments striving to cover the needs of the population for essential vaccines. A number of institutions and private structures such as Institut Pasteur Production, Connaught Laboratories, and Institut Merieux have also set up production facilities. Today's producers can be divided into two categories, i.e., local producers that produce mainly monovalent vaccines and worldwide producers with strong R&D investment programs. Local producers are located mainly in large southern hemisphere countries such as China, India, Brazil, and Indonesia as well as in eastern countries. For the most dynamic companies, international development is focused on southern hemisphere countries excluding North America and Europe. With the support international organization such as WHO, UNICEF and GAVI, alliances are now being formed and networks are being organized in an effort to ensure reliable supplies of high quality vaccines at affordable prices in developing countries. The contribution of these producers will increase for the greater benefit of the people living in the southern hemisphere.

77 FR 49776 - Southern New Mexico Resource Advisory Committee

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-17

... DEPARTMENT OF AGRICULTURE Forest Service Southern New Mexico Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Southern New Mexico Resource Advisory Committee will meet in Socorro, New Mexico. The purpose of the committee is to improve collaborative...

76 FR 46722 - Southern New Mexico Resource Advisory Committee

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-03

... DEPARTMENT OF AGRICULTURE Forest Service Southern New Mexico Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Southern New Mexico Resource Advisory Committee will meet in Socorro, New Mexico. The committee is authorized under the Secure Rural Schools and...

76 FR 16603 - Southern New Mexico Resource Advisory Committee

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-24

... DEPARTMENT OF AGRICULTURE Forest Service Southern New Mexico Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Southern New Mexico Resource Advisory Committee (RAC) will meet in Socorro, New Mexico. The committee is meeting as authorized under the Secure...

75 FR 69916 - Southern New Mexico Resource Advisory Committee

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-16

... DEPARTMENT OF AGRICULTURE Forest Service Southern New Mexico Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Southern New Mexico Resource Advisory Committee (RAC) will meet in Socorro, New Mexico. The committee is meeting as authorized under the Secure...

75 FR 39655 - Southern New Mexico Resource Advisory Committee

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-12

... DEPARTMENT OF AGRICULTURE Forest Service Southern New Mexico Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Southern New Mexico Resource Advisory Committee (RAC) will meet in Socorro, New Mexico. The committee is meeting as authorized under the Secure...

75 FR 30772 - Southern New Mexico Resource Advisory Committee

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-02

... DEPARTMENT OF AGRICULTURE Forest Service Southern New Mexico Resource Advisory Committee AGENCY: Forest Service, USDA. ACTION: Notice of meeting. SUMMARY: The Southern New Mexico Resource Advisory Committee will meet in Socorro, New Mexico. The committee is meeting as authorized under the Secure Rural...

Aggregation behavior of the southern chinch bug (Hemiptera: Blissidae)

USDA-ARS?s Scientific Manuscript database

The southern chinch bug, Blissus insularis Barber, forms dense, multigenerational aggregations in St. Augustinegrass lawns leading to grass death from sap feeding. We conducted several bioassays to better understand the signals responsible for the formation and maintenance of southern chinch bug ag...

The Spatial Concentration of Southern Whites and Argument-Based Lethal Violence

ERIC Educational Resources Information Center

Lee, Matthew R.; Shihadeh, Edward S.

2009-01-01

This analysis examines how the spatial concentration of Southern whites is associated with white argument-based lethal violence. Using a well-known measure of spatial segregation (V, the adjusted P* index) among Southern-born whites in U.S. counties in 2000, the results reveal that the spatial concentration of Southern-born whites is only…

Potential Contribution of Anammox to Nitrogen Loss from Paddy Soils in Southern China

PubMed Central

Yang, Xiao-Ru; Li, Hu; Nie, San-An; Su, Jian-Qiang; Weng, Bo-Sen; Zhu, Gui-Bing; Yao, Huai-Ying; Gilbert, Jack A.

2014-01-01

The anaerobic oxidation of ammonium (anammox) process has been observed in diverse terrestrial ecosystems, while the contribution of anammox to N2 production in paddy soils is not well documented. In this study, the anammox activity and the abundance and diversity of anammox bacteria were investigated to assess the anammox potential of 12 typical paddy soils collected in southern China. Anammox bacteria related to “Candidatus Brocadia” and “Candidatus Kuenenia” and two novel unidentified clusters were detected, with “Candidatus Brocadia” comprising 50% of the anammox population. The prevalence of the anammox was confirmed by the quantitative PCR results based on hydrazine synthase (hzsB) genes, which showed that the abundance ranged from 1.16 × 104 to 9.65 × 104 copies per gram of dry weight. The anammox rates measured by the isotope-pairing technique ranged from 0.27 to 5.25 nmol N per gram of soil per hour in these paddy soils, which contributed 0.6 to 15% to soil N2 production. It is estimated that a total loss of 2.50 × 106 Mg N per year is linked to anammox in the paddy fields in southern China, which implied that ca. 10% of the applied ammonia fertilizers is lost via the anammox process. Anammox activity was significantly correlated with the abundance of hzsB genes, soil nitrate concentration, and C/N ratio. Additionally, ammonia concentration and pH were found to be significantly correlated with the anammox bacterial structure. PMID:25416768

Archean sedimentation and tectonics in southern Africa

NASA Technical Reports Server (NTRS)

Kidd, W. S. F.

1984-01-01

Sequences in the Barberton Mountain Land greenstone belt (southern Africa) were examined to determine the nature of the sedimentary rocks, their tectonic implications, and their bearing on the present large-scale structural condition of the belt. Also assessed was whether there was evidence for a significant component of shallow-water-deposited sedimentary rocks in the parent materials of the Limpopo belt. The nature of a largehigh strain zone on the southern margin of the central Limpopo belt was examined.

Early German Plans for a Southern Observatory

NASA Astrophysics Data System (ADS)

Wolfschmidt, Gudrun

As early as the 18th and 19th centuries, French and English observers were active in South Africa. Around the beginning of the 20th century the Heidelberg astronomer Max Wolf (1863-1932) proposed a southern observatory. In 1907 Hermann Carl Vogel (1841-1907), director of the Astrophysical Observatory Potsdam, suggested a southern station in Spain. His ideas for building an observatory in Windhuk for photographing the sky and measuring the solar constant were taken over by the Göttingen astronomers. In 1910 Karl Schwarzschild (1873-1916), after having visited the observatories in America, pointed out the usefulness of an observatory in South West Africa, where it would have better weather than in Germany and also give access to the southern sky. Seeing tests were begun in 1910 by Potsdam astronomers, but WW I stopped the plans. In 1928 Erwin Finlay-Freundlich (1885-1964), inspired by the Hamburg astronomer Walter Baade (1893-1960), worked out a detailed plan for a southern observatory with a reflecting telescope, spectrographs and an astrograph with an objective prism. Paul Guthnick (1879-1947), director of the Berlin observatory, in cooperation with APO Potsdam and Hamburg, made a site survey to Africa in 1929 and found the conditions in Windhuk to be ideal. Observations were started in the 1930s by Berlin and Breslau astronomers, but were stopped by WW II. In the 1950s, astronomers from Hamburg and The Netherlands renewed the discussion in the framework of European cooperation, and this led to the founding of ESO in 1963, as is well described by Blaauw (1991). Blaauw, Adriaan: ESO's Early History. The European Southern Observatory from Concept to Reality. Garching bei München: ESO 1991.

Coatomer subunit beta 2 (COPB2), identified by label-free quantitative proteomics, regulates cell proliferation and apoptosis in human prostate carcinoma cells.

PubMed

Mi, Yuanyuan; Sun, Chuanyu; Wei, Bingbing; Sun, Feiyu; Guo, Yijun; Hu, Qingfeng; Ding, Weihong; Zhu, Lijie; Xia, Guowei

2018-01-01

Label-free quantitative proteomics has broad applications in the identification of differentially expressed proteins. Here, we applied this method to identify differentially expressed proteins (such as coatomer subunit beta 2 [COPB2]) and evaluated the functions and molecular mechanisms of these proteins in prostate cancer (PCA) cell proliferation. Proteins extracted from surgically resected PCA tissues and adjacent tissues of 3 patients were analyzed by label-free quantitative proteomics. The target protein was confirmed by bioinformatics and GEO dataset analyses. To investigate the role of the target protein in PCA, we used lentivirus-mediated small-interfering RNA (siRNA) to knockdown protein expression in the prostate carcinoma cell line, CWR22RV1 cells and assessed gene and protein expression by reverse transcription quantitative polymerase chain reaction and western blotting. CCK8 and colony formation assays were conducted to evaluate cell proliferation. Cell cycle distributions and apoptosis were assayed by flow cytometry. We selected the differentiation-related protein COPB2 as our target protein based on the results of label-free quantitative proteomics. High expression of COPB2 was found in PCA tissue and was related to poor overall survival based on a public dataset. Cell proliferation was significantly inhibited in COPB2-knockdown CWR22RV1 cells, as demonstrated by CCK8 and colony formation assays. Additionally, the apoptosis rate and percentage of cells in the G 1 phase were increased in COPB2-knockdown cells compared with those in control cells. CDK2, CDK4, and cyclin D1 were downregulated, whereas p21 Waf1/Cip1 and p27 Kip1 were upregulated, affecting the cell cycle signaling pathway. COPB2 significantly promoted CWR22RV1 cell proliferation through the cell cycle signaling pathway. Thus, silencing of COPB2 may have therapeutic applications in PCA. Copyright © 2017 Elsevier Inc. All rights reserved.

Rapid detection of rifampin-resistant clinical isolates of Mycobacterium tuberculosis by reverse dot blot hybridization.

PubMed

Guo, Qian; Yu, Yan; Zhu, Yan Ling; Zhao, Xiu Qin; Liu, Zhi Guang; Zhang, Yuan Yuan; Li, Gui Lian; Wei, Jian Hao; Wu, Yi Mou; Wan, Kang Lin

2015-01-01

A PCR-reverse dot blot hybridization (RDBH) assay was developed for rapid detection of rpoB gene mutations in 'hot mutation region' of Mycobacterium tuberculosis (M. tuberculosis). 12 oligonucleotide probes based on the wild-type and mutant genotype rpoB sequences of M. tuberculosis were designed to screen the most frequent wild-type and mutant genotypes for diagnosing RIF resistance. 300 M. tuberculosis clinical isolates were detected by RDBH, conventional drug-susceptibility testing (DST) and DNA sequencing to evaluate the RDBH assay. The sensitivity and specificity of the RDBH assay were 91.2% (165/181) and 98.3% (117/119), respectively, as compared to DST. When compared with DNA sequencing, the accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the RDBH assay were 97.7% (293/300), 98.2% (164/167), and 97.0% (129/133), respectively. Furthermore, the results indicated that the most common mutations were in codons 531 (48.6%), 526 (25.4%), 516 (8.8%), and 511 (6.6%), and the combinative mutation rate was 15 (8.3%). One and two strains of insertion and deletion were found among all strains, respectively. Our findings demonstrate that the RDBH assay is a rapid, simple and sensitive method for diagnosing RIF-resistant tuberculosis. Copyright © 2015 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

3. Oil House, Southern Pacific Railroad Carlin, northeast elevation, view ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

3. Oil House, Southern Pacific Railroad Carlin, northeast elevation, view to southwest (135mm lens). - Southern Pacific Railroad, Carlin Shops, Oil House, Foot of Sixth Street, Carlin, Elko County, NV

78 FR 14400 - Southern USA Resources, Inc., Order of Suspension of Trading

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-05

... SECURITIES AND EXCHANGE COMMISSION [File No. 500-1] Southern USA Resources, Inc., Order of... lack of current and accurate information concerning the securities of Southern USA Resources, Inc. (``Southern USA'') because of questions regarding the accuracy of publicly-disseminated information concerning...

Multilingualism in Southern Africa.

ERIC Educational Resources Information Center

Peirce, Bonny Norton; Ridge, Stanley G. M.

1997-01-01

Reviews recent research in multilingualism in Southern Africa, focusing on the role of languages in education, sociolinguistics, and language policy. Much of the research is on South Africa. Topics discussed include language of instruction in schools, teacher education, higher education, adult literacy, language contact, gender and linguistic…

Informal optics education and outreach programs in Southern California

NASA Astrophysics Data System (ADS)

Silberman, Donn M.; Cartland, Robert; Monacelli, Brian; Whitmore, Desire

2010-08-01

Over the past decade, Southern California has seen informal optics education and outreach programs grow substantially, mainly due to efforts from members of the Optical Society of Southern California (OSSC) and more recently the Optical Society of America, UC Irvine, Student Chapter. Also, the Optics Institute of Southern California (OISC) has served as a focal point for many of these programs, as an independent organization working closely with society members and other partners. This paper provides an update of these programs, including a new OSSC website that provides a new platform for significantly expanding the member participation efforts.

The southern forest resource assessment: What we learned

Treesearch

David N. Wear; John G. Greis

2004-01-01

The Southern Forest Resource Assessment was initiated in the spring of 1999 to address broad questions concerning the status, trends, and possible future of southern forests. The overall objective of the assessment was to develop a thorough and objective description of forest conditions and trends in the South, and to present it in a way that would help the public...

Application of the LEPS technique for Quantitative Precipitation Forecasting (QPF) in Southern Italy: a preliminary study

NASA Astrophysics Data System (ADS)

Federico, S.; Avolio, E.; Bellecci, C.; Colacino, M.; Walko, R. L.

2006-03-01

This paper reports preliminary results for a Limited area model Ensemble Prediction System (LEPS), based on RAMS (Regional Atmospheric Modelling System), for eight case studies of moderate-intense precipitation over Calabria, the southernmost tip of the Italian peninsula. LEPS aims to transfer the benefits of a probabilistic forecast from global to regional scales in countries where local orographic forcing is a key factor to force convection. To accomplish this task and to limit computational time in an operational implementation of LEPS, we perform a cluster analysis of ECMWF-EPS runs. Starting from the 51 members that form the ECMWF-EPS we generate five clusters. For each cluster a representative member is selected and used to provide initial and dynamic boundary conditions to RAMS, whose integrations generate LEPS. RAMS runs have 12-km horizontal resolution. To analyze the impact of enhanced horizontal resolution on quantitative precipitation forecasts, LEPS forecasts are compared to a full Brute Force (BF) ensemble. This ensemble is based on RAMS, has 36 km horizontal resolution and is generated by 51 members, nested in each ECMWF-EPS member. LEPS and BF results are compared subjectively and by objective scores. Subjective analysis is based on precipitation and probability maps of case studies whereas objective analysis is made by deterministic and probabilistic scores. Scores and maps are calculated by comparing ensemble precipitation forecasts against reports from the Calabria regional raingauge network. Results show that LEPS provided better rainfall predictions than BF for all case studies selected. This strongly suggests the importance of the enhanced horizontal resolution, compared to ensemble population, for Calabria for these cases. To further explore the impact of local physiographic features on QPF (Quantitative Precipitation Forecasting), LEPS results are also compared with a 6-km horizontal resolution deterministic forecast. Due to local and

HIV/AIDS in the Southern USA: a disproportionate epidemic.

PubMed

Reif, Susan S; Whetten, Kathryn; Wilson, Elena R; McAllaster, Carolyn; Pence, Brian W; Legrand, Sara; Gong, Wenfeng

2014-01-01

This research synthesis examined HIV/AIDS surveillance and health care financing data and reviewed relevant research literature to describe HIV epidemiology, outcomes, funding, and contributing factors to the HIV epidemic in the Southern USA with particular focus on a group of Southern states with similar demographic and disease characteristics and comparable HIV epidemics (Alabama, Georgia, Florida, Los Angeles, Mississippi, North Carolina, South Carolina, Tennessee, and Texas). These states are hereafter referred to as "targeted Southern states." Eight of the 10 states with the highest HIV diagnosis rates in 2011 were in the Southern USA; six were targeted states. Forty-nine percent of HIV diagnoses were in the South in 2011, which contains only 37% of the US population. The targeted states region had the highest HIV diagnosis rate than any other US region in 2011. The South was also found to have the highest HIV-related mortality and morbidity rates in the USA. The high levels of poverty, HIV-related stigma, and STDs found in the South, particularly in the targeted Southern states, likely contribute to greater HIV incidence and mortality. The disproportionate impact of HIV in the South, particularly among targeted states, demonstrates a critical need to improve HIV prevention and care and address factors that contribute to HIV disease in this region.

Detection of Traces of Ovalbumin and Casein in White and Red Wines by Quantitative Western Blotting.

PubMed

Meyer, Philippe; Zanetti, Simone

2015-09-01

Fining of wine with agents containing cow's milk or hen's egg white is a common and traditional procedure. In light of increasing food allergies all over the world, the presence of fining residues has been subject of intense debate. Switzerland does not make exception, and since 2009 the Federal Department of Home Affairs has modified its food regulations stating that the labels must show if traces of fining agents are present. Nevertheless, the application of this regulation is not based on an official analytical method. In this study we show that immunoblotting is an efficient technique to detect and quantify ovalbumin and casein residues in bottled wine. We showed that final filtration is an essential step to remove finings in red wine, and that overfining of white wine may result in fining residues in finished products. Finally, for the first time in Switzerland, 22 samples were taken by food safety inspectors and officially analyzed for the regional food control authority of the Canton of Vaud. These samples were allergen free, but a larger study is currently planned in collaboration with other regional authorities of Switzerland to complete these results and make a complete picture of the Swiss wine production. © 2015 Institute of Food Technologists®

Astrophysics in Southern Africa

NASA Astrophysics Data System (ADS)

Whitelock, Patricia

2008-03-01

The government of South Africa has identified astronomy as a field in which their country has a strategic advantage and is consequently investing very significantly in astronomical infrastructure. South Africa now operates a 10-m class optical telescope, the Southern African Large Telescope (SALT), and is one of two countries short listed to host the Square Kilometre Array (SKA), an ambitious international project to construct a radio telescope with a sensitivity one hundred times that of any existing telescope. The challenge now is to produce an indigenous community of users for these facilities, particularly from among the black population which was severely disadvantaged under the apartheid regime. In this paper I briefly describe the observing facilities in Southern Africa before going on to discuss the various collaborations that are allowing us to use astronomy as a tool for development, and at the same time to train a new generation of astronomers who will be well grounded in the science and linked to their colleagues internationally.

Epidemiology of Rheumatoid Arthritis in Southern Albania

PubMed Central

Koko, Vjollca; Ndrepepa, Ana; Skenderaj, Skender

2015-01-01

Objectives: The aim of this study was to assess the prevalence, incidence and the burden of rheumatoid arthritis (RA) in the Southern Albania. Material and Methods: This is an epidemiologic observational study with cross-sectional analyses of all patients with RA who lived in Southern Albania during the 1995-2011 years. The RA prevalence, incidence and disability-adjusted life years (DALY) were assessed. Results: During the 1995-2011 years, 194 patients (154 females and 40 males) with RA living in Southern Albania were identified. The prevalence of RA in 2011 was 0.25% in general population and 0.34% in adult (>14 years) population. The incidence of RA in 2011 was 0.012% (12 new cases per 100000 inhabitants) and 0.016% (16 new cases per 100000 adults). The prevalence increased (from 0.036% in 1996 to 0.25% in 2011) and the incidence did not change over the study period. The mortality was 3.2% (n=7 deaths). The DALY due to RA was 823 years per 100000 inhabitants during 1995-2011 years. Conclusion: RA in Southern Albania has a prevalence of 0.25 % and an annual incidence of 0.012% in the general population in 2011. RA was responsible for a considerable burden on the health of population during the 1995-2011 years. PMID:26236163

1. Oil House, Southern Pacific Railroad Carlin Shops, southwest facade, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

1. Oil House, Southern Pacific Railroad Carlin Shops, southwest facade, view to northeast (135mm lens). - Southern Pacific Railroad, Carlin Shops, Oil House, Foot of Sixth Street, Carlin, Elko County, NV

4. Oil House, Southern Pacific Railroad Carlin Shops, southeast elevation, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

4. Oil House, Southern Pacific Railroad Carlin Shops, southeast elevation, view to northwest (90mm lens). - Southern Pacific Railroad, Carlin Shops, Oil House, Foot of Sixth Street, Carlin, Elko County, NV

2. Oil House, Southern Pacific Railroad Carlin Shops, northwest elevation, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

2. Oil House, Southern Pacific Railroad Carlin Shops, northwest elevation, view to southeast (135mm lens). - Southern Pacific Railroad, Carlin Shops, Oil House, Foot of Sixth Street, Carlin, Elko County, NV

OVERALL VIEW OF SOUTHERN DUCTILE'S PATTERN REPAIR SHOP, SHOWING A ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

OVERALL VIEW OF SOUTHERN DUCTILE'S PATTERN REPAIR SHOP, SHOWING A SPANISH-MADE FORADIA BORING MACHINE IN THE FOREGROUND. - Southern Ductile Casting Company, Mold Making, 2217 Carolina Avenue, Bessemer, Jefferson County, AL