bluuebunny/crossref_metadata_2025_split huggingface dataset:
Microsatellite markers of water buffalo, Bubalus bubalis - development, characterisation and linkage disequilibrium studies
Authors: Nagarajan Muniyandi , Kumar Niraj , Nishanth Gopala , Haribaskar Ramachandran , Paranthaman Karthikeyani , Gupta Jalaj , Mishra Manish , Vaidhegi R , Kumar Shantanu , Ranjan Amresh K , Kumar Satish 
Published: October 2009

Abstract

Background
Microsatellite markers are highly polymorphic and widely used in genome mapping and population genetic studies in livestock species. River buffalo, Bubalus bubalis is an economically important livestock species, though only a limited number of microsatellite markers have been reported thus far in this species.

Results
In the present study, using two different approaches 571 microsatellite markers have been characterized for water buffalo. Of the 571 microsatellite markers, 498 were polymorphic with average heterozygosity of 0.51 on a panel of 24 unrelated buffalo. Fisher exact test was used to detect LD between the marker pairs. Among the 137550 pairs of marker combination, 14.58% pairs showed significant LD (P < 0.05). Further to check the suitability of these microsatellite markers to map these on a radiation hybrid map of buffalo genome, the markers were tested on Chinese hamster genomic DNA for amplification. Only seven of these markers showed amplification in Chinese hamster, and thus 564, of these can be added to the radiation hybrid map of this species.

Conclusion
The high conservation of cattle microsatellite loci in water buffalo promises the usefulness of the cattle microsatellites markers on buffalo. The polymorphic markers characterised in this study will contribute to genetic linkage and radiation hybrid mapping of water buffalo and population genetic studies.
Dual effect of a single nucleotide polymorphism in the first intron of the porcine Secreted phosphoprotein 1 gene: allele-specific binding of C/EBP beta and activation of aberrant splicing
Authors: Muráni Eduard , Ponsuksili Siriluck , Seyfert Hans-Martin , Shi Xuanming , Wimmers Klaus 
Published: October 2009

Abstract

Background
Secreted phosphoprotein 1 (SPP1 or Osteopontin, OPN) is a multifunctional matricellular glycoprotein involved in development and regeneration of skeletal muscle. Previously, we have demonstrated that porcine SPP1 shows breed-related differential mRNA expression during myogenesis. With the aim to identify putative contributing cis-regulatory DNA variation we resequenced the 5' upstream region of the gene in the respective breeds Pietrain and Duroc. We found two single nucleotide polymorphisms (SNP; [GenBank:M84121]: g.1804C>T and g.3836A>G). We focused our investigation on the SNP g.3836A>G, because in silico analysis and knowledge about the regulation of SPP1 suggested an effect of this SNP on a CCAAT/enhancer binding protein beta (C/EBPβ) responsive transcriptional enhancer.

Results
Using electrophoretic mobility shift assay we demonstrated that, similar to human SPP1, the 3' terminal end of the first intron of porcine SPP1 harbors a C/EBPβ binding site and showed that this binding site is negatively affected by the mutant G allele. Genotyping of 48 fetuses per breed revealed that the G allele segregated exclusively in Duroc fetuses with a frequency of 57 percent. Using real-time quantitative PCR we showed that, consistent with its negative effect on a transcriptional enhancer element, the G allele tends to decrease mRNA abundance of SPP1 in the fetal musculus longissimus dorsi (~1.3 fold; P ≥ 0.1).
Moreover, we showed that the SNP g.3836A>G leads to ubiquitous aberrant splicing of the first intron by generating a de novo and activating a cryptic splice acceptor site. Aberrantly spliced transcripts comprise about half of the SPP1 messages expressed by the G allele. Both aberrant splice variants differ from the native transcript by insertions in the leader sequences which do not change the reading frame of SPP1.

Conclusion
At the 3' terminal end of the first intron of the porcine SPP1 we identified a unique, dually functional SNP g.3836A>G. This SNP affects the function of the SPP1 gene at the DNA level by affecting a C/EBPβ binding site and at the RNA level by activating aberrant splicing of the first intron, and thus represents an interesting DNA-marker to study phenotypic effects of SPP1 DNA-variation.
Impact of 4.0% chlorhexidine cleansing of the umbilical cord on mortality and omphalitis among newborns of Sylhet, Bangladesh: design of a community-based cluster randomized trial
Authors: Mullany Luke C , El Arifeen Shams , Winch Peter J , Shah Rasheduzzaman , Mannan Ishtiaq , Rahman Syed M , Rahman Mohammad R , Darmstadt Gary L , Ahmed Saifuddin , Santosham Mathuram , Black Robert E , Baqui Abdullah H 
Published: October 2009

Abstract

Background
The World Health Organization recommends dry cord care for newborns but this recommendation may not be optimal in low resource settings where most births take place in an unclean environment and infections account for up to half of neonatal deaths. A previous trial in Nepal indicated that umbilical cord cleansing with 4.0% chlorhexidine could substantially reduce mortality and omphalitis risk, but policy changes await additional community-based data.

Methods
The Projahnmo Chlorhexidine study was a three-year, cluster-randomized, community-based trial to assess the impact of three cord care regimens on neonatal mortality and omphalitis. Women were recruited mid-pregnancy, received a basic package of maternal and neonatal health promotion messages, and were followed to pregnancy outcome. Newborns were visited at home by local village-based workers whose areas were randomized to either 1) single- or 2) 7-day cord cleansing with 4.0% chlorhexidine, or 3) promotion of dry cord care as recommended by WHO. All mothers received basic messages regarding hand-washing, clean cord cutting, and avoidance of harmful home-base applications to the cord. Death within 28 days and omphalitis were the primary outcomes; these were monitored directly through home visits by community health workers on days 1, 3, 6, 9, 15, and 28 after birth.

Discussion
Due to report in early 2010, the Projahnmo Chlorhexidine Study examines the impact of multiple or single chlorhexidine cleansing of the cord on neonatal mortality and omphalitis among newborns of rural Sylhet District, Bangladesh. The results of this trial will be interpreted in conjunction with a similarly designed trial previously conducted in Nepal, and will have implications for policy guidelines for optimal cord care of newborns in low resource settings in Asia.

Trial Registration
ClinicalTrials.gov (NCT00434408)
Novel immortalized human fetal liver cell line, cBAL111, has the potential to differentiate into functional hepatocytes
Authors: Deurholt Tanja , van Til Niek P , Chhatta Aniska A , ten Bloemendaal Lysbeth , Schwartlander Ruth , Payne Catherine , Plevris John N , Sauer Igor M , Chamuleau Robert AFM , Elferink Ronald PJ Oude , Seppen Jurgen , Hoekstra Ruurdtje 
Published: October 2009

Abstract

Background
A clonal cell line that combines both stable hepatic function and proliferation capacity is desirable for in vitro applications that depend on hepatic function, such as pharmacological or toxicological assays and bioartificial liver systems. Here we describe the generation and characterization of a clonal human cell line for in vitro hepatocyte applications.

Results
Cell clones derived from human fetal liver cells were immortalized by over-expression of telomerase reverse transcriptase. The resulting cell line, cBAL111, displayed hepatic functionality similar to the parental cells prior to immortalization, and did not grow in soft agar. Cell line cBAL111 expressed markers of immature hepatocytes, like glutathione S transferase and cytokeratin 19, as well as progenitor cell marker CD146 and was negative for lidocaine elimination. On the other hand, the cBAL111 cells produced urea, albumin and cytokeratin 18 and eliminated galactose. In contrast to hepatic cell lines NKNT-3 and HepG2, all hepatic functions were expressed in cBAL111, although there was considerable variation in their levels compared with primary mature hepatocytes. When transplanted in the spleen of immunodeficient mice, cBAL111 engrafted into the liver and partly differentiated into hepatocytes showing expression of human albumin and carbamoylphosphate synthetase without signs of cell fusion.

Conclusion
This novel liver cell line has the potential to differentiate into mature hepatocytes to be used for in vitro hepatocyte applications.
Human APOBEC1 cytidine deaminase edits HBV DNA
Authors: Gonzalez Minerva Cervantes , Suspène Rodolphe , Henry Michel , Guétard Denise , Wain-Hobson Simon , Vartanian Jean-Pierre 
Published: October 2009

Abstract
Retroviruses, hepadnaviruses, and some other retroelements are vulnerable to editing by single stranded DNA cytidine deaminases. Of the eleven human genes encoding such enzymes, eight have demonstrable enzymatic activity. Six of seven human APOBEC3 are able to hyperedit HBV DNA, frequently on both strands. Although human APOBEC1 (hA1) is not generally expressed in normal liver, hA1 can edit single stranded DNA in a variety of experimental assays. The possibility of ectopic expression of hA1 in vivo cannot be ruled out and interestingly, transgenic mice with A1 expressed under a liver specific promoter develop hepatocellular carcinoma. The impact of hA1 on HBV in tissue culture is varied with reports noting either reduced DNA synthesis or not, with cytidine deamination taking a low profile. We sought to examine the hA1 editing activity on replicating HBV. Using highly sensitive 3DPCR it was possible to show that hA1 edits the HBV minus DNA strand as efficiently as hA3G, considered the reference deaminase for HIV and HBV. The dinucleotide specificity of editing was unique among human cytidine deaminases providing a hallmark of use in a posteriori analyses of in vivo edited genomes. Analysis of sequences derived from the serum of two chronic carriers, indicated that hA1 explained only a small fraction of edited HBV genomes. By contrast, several human APOBEC3 deaminases were active including hA3G.
Aberrant STYK1 expression in ovarian cancer tissues and cell lines
Authors: Jackson Kesmic A , Oprea Gabriela , Handy Jeffrey , Kimbro K Sean 
Published: October 2009

AbstractBackgroundOverexpression ofSTYK1, a putative serine/threonine and tyrosine receptor protein kinase has been shown to confer tumorigenicity and metastatic potential to normal cells injected into nude mice. Mutation of a tyrosine residue in the catalytic STYK1 domain attenuates the tumorigenic potential of tumor cellsin vivo, collectively, suggesting an oncogenic role for STYK1.MethodsTo investigate the role of STYK1 expression in ovarian cancer, a panel of normal, benign, and ovarian cancer tissues was evaluated for STYK1 immunoreactivity using STYK1 antibodies. In addition, mRNA levels were measured by reverse transcription PCR and real-time PCR of estrogen receptors, GPR30 and STYK1 following treatment of ovarian cell lines with estrogen or G1, a GPR30 agonist, as well as western analysis.ResultsOur data showed higher expression of STYK1 in cancer tissues versus normal or benign. Only normal or benign, and one cancer tissue were STYK1-negative. Moreover, benign and ovarian cancer cell lines expressedSTYK1as determined by RT-PCR. Estradiol treatment of these cells resulted in up- and down-regulation ofSTYK1despite estrogen receptor status; whereas G-1, a GPR30-specific agonist, increased STYK1 mRNA levels higher than that of estradiol.ConclusionWe conclude thatSTYK1is expressed in ovarian cancer and is regulated by estrogen through a GPR30 hormone-signaling pathway, to the exclusion of estrogen receptor-alpha.
A Population‐Based and Longitudinal Study of Sexual Behavior and Multidrug‐Resistant HIV Among Patients in Clinical Care
Authors: Kozal Michael J , Amico K Rivet , Chiarella Jennifer , Cornman Deborah , Fisher William , Fisher Jeffrey , Friedland Gerald 
Published: October 2009

BackgroundPopulation‐based and longitudinal information regarding sexual risk behavior among patients with multidrug resistant (MDR) HIV and their sexual partners is of great public health and clinical importance.ObjectiveTo characterize the HIV sexual risk behaviors of patients with and without drug‐resistant HIV in the clinical care setting over time.Measurements393 HIV‐positive patients completed questionnaires of self‐reported sexual risk behaviors at approximate 6‐month intervals extending over 24 months. HIV viral load and genotypic drug resistance obtained during the same time points were matched to the behavioral data. Multidrug resistance was defined as having resistance to 2 or 3 antiretroviral (ARV) drug classes.ResultsIn serial cross‐sectional analyses, 393 patients (44% female and 79% heterosexual) contributed 919 matched behavioral and virologic results over the 24 months of data collection. Of these, 250 patients (64%) reported having sex during at least 1 survey period resulting in greater than 10,000 sexual events with more than 1000 partners. Unprotected sexual behavior was reported by 45% of sexually active patients, resulting in 34% of all sex events that exposed 29% of all partners. Of these patients with unprotected sexual events, 31% had HIV drug resistance 11.6% with resistance to 2 classes of ARVs (2‐class), and 1.8% with 3‐class ARV resistance at the time of a sexual risk event. Close to 1000 or 28% of all unprotected sexual events involved resistant strains (11% of these with resistance to 2 classes and 0.2% with 3‐class resistance, exposing 20% of unprotected sexual partners to resistant HIV (8% to 2‐class and 0.6% to 3‐class resistance).In longitudinal analysis among the 78 patients who reported a cumulative total of 12 months of sexual history and had resistance testing, 38% reported engaging in unprotected sexual behavior. There was substantial and complex variation in the distribution of unprotected sexual events and in the detection of resistance over time.ConclusionIn this study of HIV sexual risk and resistance over time among HIV‐infected patients in clinical care, a substantial proportion engaged in unprotected sex and had drug‐resistant HIV, frequently exposing partners to 1‐ or 2‐class resistant HIV strains. However, relatively few exposures involved 3‐class resistance. The dynamics of sexual risk behavior and HIV drug resistance are complex and vary over time and urgently require both general and targeted interventions to reduce transmission of resistant HIV.
One year mortality of patients treated with an emergency department based early goal directed therapy protocol for severe sepsis and septic shock: a before and after study
Authors: Puskarich Michael A , Marchick Michael R , Kline Jeffrey A , Steuerwald Michael T , Jones Alan E 
Published: October 2009

Abstract
Introduction
Early structured resuscitation of severe sepsis has been suggested to improve short term mortality; however, no previous study has examined the long-term effect of this therapy. We sought to determine one year outcomes associated with implementation of early goal directed therapy (EGDT) in the emergency department (ED) care of sepsis.

Methods
We performed a longitudinal analysis of a prospective before and after study conducted at a large urban ED. Adult patients were enrolled if they had suspected infection, 2 or more systemic inflammatory response criteria, and either systolic blood pressure (SBP) <90 mmHg after a fluid bolus or lactate >4 mM. Exclusion criteria were: age <18 years, no aggressive care desired, or need for immediate surgery. Clinical and outcomes data were prospectively collected on consecutive eligible patients for 1 year before and 2 years after implementing EGDT. Patients in the pre-implementation phase received non-protocolized care at attending physician discretion. The primary outcome was mortality at one year.

Results
285 subjects, 79 in the pre- and 206 in the post-implementation phases, were enrolled. Compared to pre-implementation, post-implementation subjects had a significantly lower ED SBP (72 vs. 85 mm Hg, P < 0.001) and higher sequential organ failure assessment score (7 vs. 5, P = 0.0004). The primary outcome of 1 year mortality was observed in 39/79 (49%) pre-implementation subjects and 77/206 (37%) post-implementation subjects (difference 12%; P = 0.04).

Conclusions
Implementation of EGDT for the treatment of ED patients with severe sepsis and septic shock was associated with significantly lower mortality at one year.
Monocyte heterogeneity underlying phenotypic changes in monocytes according to SIV disease stage
Authors: Kim Woong-Ki , Sun Yue , Do Hien , Autissier Patrick , Halpern Elkan F , Piatak Michael , Lifson Jeffrey D , Burdo Tricia H , McGrath Michael S , Williams Kenneth 
Published: October 2009

Abstract
Expansion of two monocyte subpopulations expressing CD16 and their correlation with viral load are observed in this cross-sectional study of SIV-infected macaques.
Infection by HIV is associated with the expansion of monocytes expressing CD16 antigens, but the significance of this in HIV pathogenesis is largely unknown. In rhesus macaques, at least three subpopulations of blood monocytes were identified based on their expression of CD14 and CD16: CD14highCD16−, CD14highCD16low, and CD14lowCD16high. The phenotypes and functions of these subpopulations, including CD16+ monocytes, were investigated in normal, uninfected rhesus macaques and macaques that were infected with SIV or chimeric SHIV. To assess whether these different monocyte subpopulations expand or contract in AIDS pathogenesis, we conducted a cross-sectional study of 54 SIV- or SHIV-infected macaques and 48 uninfected controls. The absolute numbers of monocyte populations were examined in acutely infected animals, chronically infected animals with no detectable plasma virus RNA, chronically infected animals with detectable plasma virus RNA, and animals that died with AIDS. The absolute numbers of CD14highCD16low and CD14lowCD16high monocytes were elevated significantly in acutely infected animals and chronically infected animals with detectable plasma virus RNA compared with uninfected controls. Moreover, a significant, positive correlation was evident between the number of CD14highCD16low or CD14lowCD16high monocytes and plasma viral load in the infected cohort. These data show the dynamic changes of blood monocytes, most notably, CD14highCD16low monocytes during lentiviral infection, which are specific to disease stage.
Antigen-induced Erk1/2 activation regulates Ets-1-mediated sensitization of CD8+ T cells for IL-12 responses
Authors: Li Qingsheng , Eppolito Cheryl , Odunsi Kunle , Shrikant Protul A 
Published: October 2009

Abstract
The molecular basis of collaboration between instructions that program CD8+ T cells for long-term responses.
The presence of IL-12 during antigen stimulation instructs naive CD8+ T cells for long-term effector responses, but their mechanisms of collaboration are not understood completely. Herein, we report that CD8+ T cells (OT-I T cells) stimulated with antigen for a longer duration show enhanced sensitization to IL-12 as a result of Erk1/2-dependent, increased Ets-1 phosphorylation and subsequent increases in IL-12Rβ2 expression. Correspondingly, naive OT-I T cells stimulated by antigen for a longer duration in the presence of IL-12, irrespective of frequency of APCs, show robust effector maturation and mount long-term antigen-recall responses upon adoptive transfer. These results identify the role of antigen strength-dependent Erk1/2 activation for Ets-1-mediated collaboration with IL-12 in CD8+ T cells.
Immunopathogenesis of polymicrobial otitis media
Authors: Bakaletz Lauren O 
Published: October 2009

Abstract
The synergistic relationship between URT viruses and bacteria in OM pathogenesis is not fully understood, but overall is predicated on viral impairment of airway defenses.
OM, or inflammation of the middle ear, is a highly prevalent infection in children worldwide. OM is a multifactorial disease with multiple risk factors, including preceding or concurrent viral URT infection. Hence, OM is also a polymicrobial disease. The mechanisms by which viruses predispose to bacterial OM are replete; however, all are predicated on the general principle of compromise of primary host airway defenses. Thus, despite an as-yet incomplete understanding of the molecular mechanisms involved in bacterial superinfection of a virus-compromised respiratory tract, the URT viruses are known to induce histopathology of airway mucosal epithelium, up-regulate expression of eukaryotic receptors used for bacterial adherence, alter the biochemical and rheological properties of airway mucus, and affect innate and acquired host immune functions, among others. Although discussed here in the context of OM, during preceding or concurrent viral infection of the human respiratory tract, viral impairment of airway defenses and the resulting predisposition to subsequent bacterial coinfection are also known to be operational in the mid and lower airway as well.
Signaling through TLR7 enhances the immunosuppressive activity of murine CD4+CD25+ T regulatory cells
Authors: Forward Nicholas A , Furlong Suzanne J , Yang Yongjun , Lin Tong-Jun , Hoskin David W 
Published: October 2009

Abstract
In vitro and in vivo stimulation of the TLR7 signaling pathway of T regulatory cells causes increased IL-2 responsiveness and enhanced suppressor function.
Although signaling through certain TLRs is known to modulate the function of T lymphocytes, the effect of TLR7 stimulation on CD4+CD25+ Treg cell activity has not yet been elucidated. In this study, we show that mouse CD4+CD25+ Treg cells express TLR7 mRNA and protein. We therefore used the TLR7 agonists imiquimod, gardiquimod, and single-stranded poly(U) to show that TLR7 stimulation enhanced the ability of murine Treg cells to suppress anti-CD3/anti-CD28 mAb-coated bead-stimulated proliferation of syngeneic CD4+CD25– Tresp cells. In contrast, imiquimod failed to enhance the suppressor function of Treg cells from mice deficient in the MyD88 adaptor protein involved in TLR7 and other TLR signal transduction. Imiquimod increased murine Treg cell-mediated suppression of Tresp cell proliferation induced by anti-TCRβ mAb in the presence of syngeneic BMDCs, and Treg cells from gardiquimod-treated mice exhibited enhanced in vitro suppressor function. Moreover, levels of Tresp cell-secreted IL-2 and IFN-γ were reduced further in the presence of Treg cells plus imiquimod in comparison with Treg cells alone. In addition, imiquimod treatment increased CD25 expression by Treg cells and caused exogenous IL-2 to enhance Treg cell suppressor function. Furthermore, combined treatment with imiquimod and IL-2 increased Foxp3 expression by Treg cells. Collectively, these findings suggest that TLR7 signaling enhanced the suppressor function of Treg cells by sensitizing Treg cells to IL-2-induced activation. We speculate that TLR7-stimulated enhancement of Treg cell suppressor function may modulate host T cell responses against ssRNA viruses.