bluuebunny/crossref_metadata_2025 huggingface dataset:
The character of anti-HCV T cell responses differs between spontaneous and treatment-induced viral clearance s
Authors: Barrett L , Hirsch G , Gallant M , Howley C , Peltekian K , Grant M
Published: May 2017

Introduction: Hepatitis C virus (HCV) is a common infection with approximately 170 million individuals infected worldwide. The majority of exposed individuals develop chronic infection, with viral clearance being the exception to the rule. Even with antiviral therapy, only 50-75% of treated individuals become HCV RNA negative. The role of the adaptive immune response, and particularly T cells, has been well established in other viral infections but its function in HCV infection is unclear, making it difficult to develop either prophylactic or therapeutic vaccine strategies.

Objectives: Compare the breadth, specificity, and magnitude of anti-HCV T cell responses in individuals who are chronically HCV-infected with those who clear HCV spontaneously or after antiviral treatment.

Methods: We assessed T cell responses in 3 groups of HCV exposed individuals: 18 people who were chronically HCV infected (anti-HCV antibody and HCV RNA positive), 5 who were exposed to HCV but cleared the virus without treatment (antibody positive, RNA negative), and 17 who responded to anti-HCV treatment (antibody positive but RNA negative after treatment). CD4+ T cell responses to HCV and recall antigens were measured by standard 3H-thymidine proliferation assay, and CD8+ T cell responses were evaluated by incubating peripheral blood mononuclear cells (PBMC) overnight with 15-mer peptides spanning the entire HCV-1a genome. The number of HCV-specific interferon-gamma (IFN-g)-producing cells per million PBMC was estimated by ELISPOT.

Results: Anti-HCV CD4+ T cell responses were uncommon, and of low magnitude in all groups. Responses were only to peptides in the core region. The overall breadth of the anti-HCV CD8+ response was greatest in treated responders (average 43/44 peptide pools tested positive) and least in those who failed treatment (average 11/44). All clearers had responses to the NS5 region. The magnitude of the CD8+ response tended to be highest in the treated responders, but there was no significant difference between groups.

Conclusions: The character of the anti-HCV T cell response differs not just between chronic infection and clearance, but also between spontaneous versus treatment-associated HCV clearance. This suggests that it is likely a combination of T cell breadth, magnitude and specificity that results in viral clearance. Based on these results, development of therapeutic and prophylactic vaccine strategies may need to target different T cell responses.
Through the looking-glass: Objectivity, interpretation, and the construction of social kinds as emerging issues in research ethicss
Authors: Berkhout S G , Tyndall M W , Anderson S
Published: May 2017

Background: Access to the means of knowledge production is severely limited for many individuals who are socially, politically, and economically disadvantaged. Of particular concern, largely overlooked in bioethics discourse, is the manner by which surveillance, identification, and classification contribute to the creation of particular populations based on harmful social identities, and the subsequent stigmatization of the individuals subject to analysis.

Methods: Drawing from a range of philosophical and sociological literature, including Hans-Georg Gadamer, Charles Taylor, Bruno Latour, and Deborah Lupton, as well as public health discourse, this paper gives an analysis of a challenging metaphysical and ethical problem related to research practices and public health interventions in vulnerable communities. The themes of objectivism and stigmatization are illuminated by focusing on research related to HIV infection among survival sex workers in Vancouver’s downtown eastside.

Results: Reductionist models of human behaviour presuppose a clear distinction between description and evaluation, contributing to an understanding of social reality as structured by objective, base level data, and misconstruing modes of social relations as individual action. In the context of HIV risk, an ethical problem emerges: describing individual actions as “data,” or “fact,” serves to construct and reinforce transgressive social identities. Given that stigmatization is itself part of the context of adverse health outcomes and HIV infection, practices relying on methods that presume representational authority and fail to critically interpret human action are ethically problematic. Resolving difficulties within the methodological underpinnings of public health and HIV behavioural research requires an understanding of many social science methods as hermeneutical, while enabling the contribution of disparate standpoints to the production of scientific knowledge.

Conclusions: The implications of presumed objectivity and representational authority demand a reconsideration of how research in vulnerable communities might be conducted, such that it produces results that are scientifically rigorous and socially responsible.
Rapid electrical pacing of cardiomyocytes alters the behavior of cardiac fibroblasts: Implications for atrial fibrillations
Authors: Burstein Brett , Qi Xiao-Yan , Calderone Angelino , Nattel Stanley
Published: May 2017

Background: Atrial fibrillation (AF) produces atrial tachycardia-related changes in atrial electrophysiology, providing a possible explanation for the progressive nature of the arrhythmia. Structural remodeling results from ventricular dysfunction due to a rapid response to AF, but whether AF itself can cause atrial fibrosis remains unclear. This study investigated the hypothesis that rapid atrial cardiomyocyte activation produces factors that influence atrial fibroblast proliferation and secretory functions.

Methods: Cultured canine atrial fibroblasts were treated with medium from rapidly-paced atrial cardiomyocytes (group 5Hz), non-paced cardiomyocytes (group NP) and cardiomyocyte pacing medium alone (group Ø), and analyzed by [3H]thymidine incorporation, Western-blot and real-time RT-PCR.

Results: Rapidly-paced cardiomyocyte-conditioned medium had a potent anti-proliferative effect, reducing [3H]thymidine uptake compared to non-paced cardiomyocyte-conditioned medium and medium alone (2,143±1,490** vs. 15,870±3,343 and 16,440±4,193 respectively, **P < 0.01 vs. both NP and Ø). Conditioned medium from rapidly-paced cardiomyocytes increased αSMA protein reflecting an activated myofibroblast phenotype (1.62±0.20** vs. 1.08±0.26 and 1.01±0.24), collagen-1 (2.41±0.22* vs. 1.49±0.33 and 1.10±0.19, *P < 0.05 both NP and Ø) and fibronectin-1 (3.05±0.28* vs. 1.80±0.45 and 1.20±0.19) mRNA expression compared to control cells. Treatment of the conditioned medium with the AT1 receptor blocker valsartan, partially attenuated the pacing-induced αSMA increase but had no effect on fibroblast proliferation. Atrial tissue from 1-week atrial-tachypaced dogs with AV block/ventricular pacing to control ventricular rate similarly showed upregulation of collagen-1 (2.6±0.5 vs. 0.7±0.3, P < 0.01) and fibronectin-1 (4.9±0.6 vs. 1.1±0.4, P < 0.001) mRNA versus unpaced shams. Conclusion: Rapidly-paced cardiomyocytes release substances that profoundly alter cardiac fibroblast function and induce an activated myofibroblast phenotype that is reflected by altered ECM-gene expression in vivo. These findings are consistent with recent observations that AF per se causes ECM remodeling and may have important consequences for understanding and preventing the mechanisms underlying AF progression.
The metalloproteinase ADAM10 is required for retinal ganglion cell axon guidance in the developing visual systems
Authors: Chen Y. Y. , Hehr C. L. , Atkinson-Leadbeater K. , Hocking J. C. , McFarlane S.
Published: May 2017

Background: The growth cone interprets cues in its environment in order to reach its target. We want to identify molecules that regulate growth cone behaviour in the developing embryo. We investigated the role of A disintegrin and metalloproteinase 10 (ADAM10) in axon guidance in the developing visual system of African frog, Xenopus laevis.

Methods: We first examined the expression patterns of adam10 mRNA by in situ hybridization. We then exposed the developing optic tract to an ADAM10 inhibitor, GI254023X, in vivo. Lastly, we inhibited ADAM10 function in diencephalic neuroepithelial cells (through which retinal ganglion cell (RGC) axons extend) or RGCs by electroporating or transfecting an ADAM10 dominant negative (dn-adam10).

Results: We show that adam10 mRNA is expressed in the dorsal neuroepithelium over the time RGC axons extend towards their target, the optic tectum. Second, pharmacological inhibition of ADAM10 in an in vivo exposed brain preparation causes the failure of RGC axons to recognize their target at low concentrations (0.5, 1 μM), and the failure of the axons to make a caudal turn in the mid-diencephalon at higher concentration (5 μM). Thus, ADAM10 function is required for RGC axon guidance at two key guidance decisions. Finally, molecular inhibition of ADAM10 function by electroporating dn-adam10 in the brain neuroepithelium causes defects in RGC axon target recognition (57%) and/or defects in caudal turn (12%), as seen with the pharmacological inhibitor. In contrast, molecular inhibition of ADAM10 within the RGC axons has no effect. Conclusions: These data argue strongly that ADAM10 acts cell non-autonomously within the neuroepithelium to regulate the guidance of RGC axons. This study shows for the first time that a metalloproteinase acts in a cell non-autonomous fashion to direct vertebrate axon growth. It will provide important insights into candidate molecules that could be used to reform nerve connections if destroyed because of injury or disease.

References

Hattori M, Osterfield M, Flanagan JG. Regulated cleavage of a contact-mediated axon repellent. Science 2000; 289(5483):1360-5.

Janes PW, Saha N, Barton WA, Kolev MV, Wimmer-Kleikamp SH, Nievergall E, Blobel CP, Himanen JP, Lackmann M, Nikolov DB. Adam meets Eph: an ADAM substrate recognition module acts as a molecular switch for ephrin cleavage in trans. Cell 2005; 123(2):291-304.

Pan D, Rubin GM. Kuzbanian controls proteolytic processing of Notch and mediates lateral inhibition during Drosophila and vertebrate neurogenesis. Cell 1997; 90(2):271-80.
Innate inflammatory and phagocytic responses to Plasmodium falciparum: linked processes or molecularly discrete pathways?s
Authors: Erdman Laura , Cosio Gabriela , Patel Samir N. , Grinstein Sergio , Kain Kevin C.
Published: May 2017

Background: Effective innate immune responses are important for control of malaria blood-stage infection and in preventing progression to severe malaria in non-immune individuals. Key innate defenses include a tightly regulated inflammatory response and host clearance of parasitized erythrocytes (PE). Pattern recognition receptors on macrophages mediate these processes: parasite glycosylphosphatidylinositol (GPI) activates TLR2 to induce inflammation – an excess of which is associated with severe malaria – while scavenger receptor CD36 mediates non-opsonic uptake of PEs. Both pathways are potential therapeutic targets, but it is unclear whether they are interdependent. Findings in other systems implicate CD36 in inflammation and TLR2 in phagocytosis, and recent evidence indicates that CD36 and TLR2 can directly cooperate.

Methods: We investigated whether the innate inflammatory and phagocytic responses of macrophages to P. falciparum are separable: does CD36-mediated PE internalization have inflammatory outcomes, and does TLR2 regulate PE uptake? CD36-mediated endocytosis failed to induce TNFα production. As a more representative model of innate PE uptake, α-CD36 EBABs (Erythrocyte-Biotin-Avidin-Biotinylated antibody) were generated; macrophages internalized EBABs in a CD36-specific manner via a signaling pathway similar to that of PE uptake.

Results: Compared to controls, neither PE nor EBAB internalization induced TNFα release, indicating that the inflammatory consequences of CD36 engagement are ligand dependent. Regarding TLR2 regulation of PE uptake, wild type and Tlr2-/- macrophages showed no differences in EBAB or PE uptake. Pre-treatment of macrophages with P. falciparum GPI enhanced EBAB internalization, but this effect was CD36-independent and generalizable to other TLR ligands.

Conclusions: These results suggest that innate inflammatory and phagocytic responses of macrophages to malaria are discrete. Thus, therapeutic augmentation of CD36-mediated PE uptake should not exacerbate inflammation, nor should inhibition of the TLR2 pathway compromise CD36-mediated PE clearance. The role of TLR-enhanced internalization in malaria will be further examined.
Differential expression of type I interferon genes in plasmacytoid dendritic cells from HIV-infected patientss
Authors: Hyrcza Martin D. , Der Sandy S. , Ostrowski Mario
Published: May 2017

Background: Plasmacytoid dendritic cells (pDCs) are the most potent producers of type I interferons (IFN). Human genome contains thirteen IFN alpha genes and one IFN beta gene. Research in mice suggests that different IFNs are induced by different stimuli, but whether this is true in human cells is unknown. Patients with HIV-1 infection show chronic interferon response in peripheral T cells, which caused us to analyze the induction of the IFN alpha genes in their dendritic cells.

Methods: Uninfected, acutely infected and long-term non-progressive donors were leukopheresed, following which pDCs were isolated by negative depletion. The dendritic cells were then treated for with one of the following: influenza virus, sendai virus, HIV virus, CpG DNA, imiquimod, or media alone, and the cells’ RNA was analysed by real time qPCR for changes in the RNA levels of four IFN alpha genes: α1, α2, α7, α8, as well as IFN beta.

Results: Final results were not available at the time of abstract deposition.
Genetic characterization of two autosomal recessive disorders, Majewski-like and cerebral atrophy syndromes
Authors: Lahiry P , Robinson J F , Siu V , Puffenberger E G , Strauss K A , Hegele R A , Rupar C A
Published: May 2017

Introduction: We recently identified two lethal recessive disorders segregating within the same Old Order Amish pedigree. The first disorder, Majewski-like syndrome (MLS), has features that overlap with both Majewski short rib-polydactyly syndrome and hydrolethalus syndrome. MLS is a lethal multi-system disorder that affects the development of the brain, adrenal glands, pituitary gland and bone. The second disorder, cerebral atrophy syndrome (CAS), is characterized by progressive and global loss of brain tissue. Affected children present early in life with microcephaly, seizures, and psychomotor retardation, and possess distinctive MRI findings.

The objective of this study was to identify the genetic bases of these disorders to provide prompt and reliable diagnosis for families.

Methods: Assuming recessive inheritance and mutation homogeneity (autozygosity), we used Affymetrix 10,000-single nucleotide polymorphism (10K-SNP) to genotype all affected individuals and identify candidate regions. SNP data were analyzed using Agilent GT autozygosity mapping software. LOD scores were used to identify candidate regions, and genes within these regions were prioritized for sequencing.

Results and Conclusion: Because the Ontario Anabaptist population is relatively small, genetically isolated, and historically young, we were able to robustly map candidate regions using relatively low marker density and only a few affected individuals. Our preliminary data is consistent with the clinical observation that MLS and CAS segregate independently, as recessive conditions, within the pedigree. Thus far, we have sequenced 12 genes within the MLS locus, all of which were normal. For CAS, autozygosity mapping yielded two loci with comparable linkage scores, one of which contained no observable mutations. Once the causative mutations have been identified for MLS and CAS, we intend to study their population frequencies and also to pursue in vitro studies of gene and protein functions.
Response of human osteoblast-like cells to fluid flow shear: A potential role for the microtubule network and primary ciliums
Authors: Myers Kenneth A. , Rattner Jerome B. , Shrive Nigel G. , Hart David A.
Published: May 2017

Introduction: A limited understanding of the cellular mechanisms governing bone mechanotransduction has inhibited the development of clinical treatments for a variety of bone disorders, including osteoporosis, osteoarthritis and microgravity-associated bone atrophy. The cytoskeleton is thought to play a role in cellular mechanotransduction, however the exact mechanism in bone cells has not yet been clearly elucidated. Studies involving cytoskeletal inhibitors have not generally considered secondary effects on cellular organelles such as the primary cilia. These cellular projections could account for the disparity between shear stresses predicted to occur in vivo and the minimum threshold of membrane deformation required to elicit a cellular response in vitro.

Methods: MG-63 (human osteoblast-like) cells were cultured in vitro. Cultures were exposed to intermittent cyclic fluid flow shear stress (1 Pa amplitude), for 8 or 12 hrs. Some cultures were loaded in the presence of nocodazole (a microtubule inhibitor) or cytochalasin D (an actin filament inhibitor). The cellular response was analyzed through RT-PCR assessment of messenger RNA levels for specific molecules related to matrix metabolism. The effects of drug treatments on cytoskeletal disorganization and the primary cilia were assessed with immunocytochemistry and electron microscopy.

Results: In untreated cultures, shear stress was associated with significant increases in mRNA levels for collagen I and matrix metalloproteinases 1 and 3, for both time points assessed. These increases were maintained in cultures loaded in the presence of cytochalasin D, but were almost completely abrogated in nocodazole-treated cultures. Cytoskeletal inhibitors exerted some dose-dependent effects on length and structure of primary cilia in MG-63 cells.

Conclusions: The microtubule network appears to be necessary for some shear-induced responses of osteoblast-like cells. MG-63 cells possess primary cilia, organelles that could amplify fluid flow shear, accounting for some apparent contradictions between studies related to osteoblast mechanosensitivity. Since these structures are composed of microtubules, the observation that microtubule disruptors inhibit the shear response of osteoblast-like cells suggests the primary cilium may have a role in osteoblast mechanotransduction. The effects of cytoskeletal inhibitors on cilium structure may explain the conflicting results of earlier mechanotransduction studies.
Examination of the dynamics of global DNA methylation pattern establishment during spermatogenesiss
Authors: Niles Kirsten , La Salle Sophie , Oakes Christopher , Trasler Jacquetta
Published: May 2017

Background: DNA methylation is an epigenetic modification involved in gene expression, genome stability, and genomic imprinting. In the male, methylation patterns are initially erased in primordial germ cells (PGCs) as they enter the gonadal ridge; methylation patterns are then acquired on CpG dinucleotides during gametogenesis. Correct pattern establishment is essential for normal spermatogenesis. To date, the characterization and timing of methylation pattern acquisition in PGCs has been described using a limited number of specific gene loci. This study aimed to describe DNA methylation pattern establishment dynamics during male gametogenesis through global methylation profiling techniques in a mouse model.

Methods: Using a chromosome based approach, primers were designed for 24 regions spanning chromosome 9; intergenic, non-repeat, non-CpG island sequences were chosen for study based on previous evidence that these types of sequences are targets for testis-specific methylation events. The percent methylation was determined in each region by quantitative analysis of DNA methylation using real-time PCR (qAMP). The germ cell-specific pattern was determined by comparing methylation between spermatozoa and liver. To examine methylation in developing germ cells, spermatogonia from 2 day- and 6 day-old Oct4-GFP (green fluorescent protein) mice were isolated using fluorescence activated cell sorting.

Results: As compared to liver, four loci were hypomethylated and five loci were hypermethylated in spermatozoa, supporting previous results indicating a unique methylation pattern in male germ cells. Only one region was hypomethylated and no regions were hypermethylated in day 6 spermatogonia as compared to mature spermatozoa, signifying that the bulk of DNA methylation is established prior to type A spermatogonia. The methylation in day 2 spermatogonia, germ cells that are just commencing mitosis, revealed differences of 15-20% compared to day 6 spermatogonia at five regions indicating that the most crucial phase of DNA methylation acquisition occurs prenatally.

Conclusion: Together, these studies provide further evidence that germ cell methylation patterns differ from those in somatic tissues and suggest that much of methylation at intergenic sites is acquired during prenatal germ cell development. (Supported by CIHR)
Stepping up regulatory mechanisms of GLUT4 traffic in L6 skeletal muscle cellss
Authors: Randhawa V. K. , Talior-Volodarsky I. , Klip A.
Published: May 2017

Insulin increases glucose uptake into muscle and fat by enhancing GLUT4 glucose transporter externalization; a process requiring input from Akt and actin. Downstream of phosphatidylinositol-3-kinase, insulin signaling bifurcates into Akt and actin activating arms. Akt-mediated phosphorylation of the Rab-GAP AS160 is required for gain in surface GLUT4 by insulin. However, little is known of the mechanism(s) by which AS160 and/or actin dynamics modulate GLUT4 traffic in muscle. We recently showed that GLUT4 arrival and/or fusion can be regulated by insulin signaling molecules and phospholipids. Using ‘rounded up’ L6 myoblasts stably expressing GLUT4myc, we find that transient expression of a non-phosphorylatable mutant of AS160 (AS160-4P) abrogates the surface fusion of GLUT4myc and partially reduces its sub-membranous accumulation. In contrast, tetanus toxin-mediated cleavage of VAMP2 inhibits GLUT4myc fusion but not arrival to the plasma membrane. Conversely, disrupting actin dynamics with Latrunculin B or silencing expression of a cytoskeletal protein a-actinin4 precludes the insulin-induced cortical build-up of GLUT4myc. These data suggest that AS160 and actin dynamics impinge on distinct stages of insulin-regulated GLUT4 traffic: AS160 may contribute to peripheral retention and is essential for GLUT4myc vesicle docking/fusion. It will be interesting to note which Rabs facilitate these AS160-dependent events. Actin dynamics instead may allow GLUT4 vesicle movement to the cell surface and/or its retention, presumably via cortical anchoring mechanisms involving a-actinin4, whilst VAMP2 has a major role in GLUT4 vesicle fusion. Indeed, defects in AS160 phosphorylation and actin dynamics are associated with insulin resistant states. Thus, discerning which steps of GLUT4 traffic are modulated by these inputs may help elucidate strategies to bypass insulin resistance.
Development of a thermally responsive peptide for sustained deliver of solyble TNF receptor II to attenuate inflammatory events associated with radiculopathys
Authors: Shamji Mohammed F. , Ghodsizadeh Odelia , Friedman Allan H. , Richardson William J. , Chilkoti Ashutosh , Setton Lori A.
Published: May 2017

Background: Tumor necrosis factor alpha (TNFα) is a cytokine that may mediate inflammatory histopathology of the dorsal root ganglion following lumbar disc herniation.1 Soluble TNF receptor II (sTNFRII) competitively binds TNFa with clinical value for painful radiculopathy.2 Bioactive peptides expressed with elastin-like polypeptides (ELP) fusion partners gain a thermally responsive domain, by which they can undergo hydrophobic collapse and separate from solution to aggregate at physiological temperatures.3 Protein release from such a depot may locally sustain drug presence, an effect demonstrated for non-fusion ELP after intra-articular injection.4

Methods: We expressed sTNFRII fused to ELP to demonstrate potential bidomain functionality.

Protein Expression. A gene encoding ELP-(VPGVG)60 was subcloned adjacent to the sTNFRII and transformed into E.coli for expression.5 Protein Safety. Endotoxin content of purified fusion protein was evaluated using a limulus amebocyte lysate endpoint assay and compared to non-fusion ELP using a two-tailed Student’s t-test. Thermal Responsiveness. Dynamic light scattering evaluated the inverse thermal phase transition behaviour of ELP-sTNFRII, and absorbance spectrophotometry quantified the in vitro depot release at 37°C. Fusion Domain Function. Anti-TNFα bioactivity was assessed by the in vitro inhibition of TNFα-induced glutamate production by microglia. Single-factor ANOVA analyzed treatment differences for ELP-sTNFRII, commercial sTNFRII (positive control), and non-fusion ELP (negative control).

A 44 kDa recombinant fusion protein was expressed from E. coli and purified by inverse transition cycling.

Results: Measured endotoxin content for ELP-sTNFRII was comparable to ELP alone (p < 0.01), well below FDA levels for biomedical implants. The fusion protein underwent a thermally-induced phase transition and formed observable aggregates of ~240 nm upon heating to physiological temperatures (Tt = 32°C). Slow release was observed from this depot with a time constant of 21 ± 3 hours. The fusion protein demonstrated anti-TNFα activity in vitro by attenuating TNFα-induced microglial glutamate production, albeit requiring a greater concentration than the free antagonist to achieve the same effect.(p < 0.01).

Conclusion: Fusion of a sTNFRII protein to an ELP can serve to generate a thermally-induced drug depot that may sustain anti-cytokine activity of agents delivered locally to a nerve region. Further directions may involve studying in vivo biodistribution after perineural delivery of ELP and in vivo disease modifying activity of this agent.
Inflammatory cytokines and S-100b protein in patients with hepatitis C infection and cryoglobulinemias
Authors: Falasca Katia , Mancino Paola , Ucciferri Claudio , Dalessandro Margherita , Zingariello Pompea , Lattanzio Franca M. , Petrarca Claudia , Martinotti Stefano , Pizzigallo Eligio , Conti Pio , Vecchiet Jacopo
Published: May 2017

Objective: To investigate a predictive role for the protein S-100b and serum circulating levels of Th1/Th2 cytokines in patients with chronic hepatitis C virus (HCV) infection with and without mixed cryoglobulinemia (MC).

Methods: Sixty chronically HCV-infected patients were divided into two groups: 30 with and 30 without MC. Patients with MC presented detectable mixed cryoglobulins and clinical weakness, purpura and arthralgias. HCV-RNA and genotype, serum levels of cryoglobulins, principal hepatic indexes and levels of IL-6, IL-18 and S-100b protein were evaluated. Twenty uninfected healthy subjects were a control group to evaluate serum levels of S-100b protein.

Results: IL-6 and IL-18 serum levels were higher in the MC+ group than the MC- group (8.7 ± 4.5 pg/mL versus 4.6 ± 2.3 pg/mL P < 0.0001 and 743.5 ± 128.2 pg/mL versus 578.5 ± 296.5 pg/mL P < 0.001 respectively). S-100b serum levels were higher in HCV+ with MC (0.23 ± 0.07 microg/L) respect to HCV+ patients without MC (0.17 ± 0.05 microg/L, P < 0.0001) and were statistically higher than in the control group (0.08 ± 0.03 microg/L, P < 0.0001 and P < 0.0001, respectively). A positive correlation was shown between serum levels of S-100b protein and levels of cryoglobulins in the group of HCV+ patients MC+ (r=0.72 and P < 0.0001).

Conclusion: HCV patients with MC have a worse inflammatory condition than those without MC. Moreover, S-100b protein seems to be a sensitive marker of endothelial and tissue damage in chronic HCV hepatitis with cryoglobulinemia.
Measurement of cortisol in human Hair as a biomarker of systemic exposure
Authors: Sauvé Brittany , Koren Gideon , Walsh Grace , Tokmakejian Sonya , Van Uum Stan HM
Published: May 2017

Purpose: Current methods for measuring long-term endogenous production of cortisol can be challenging due to the need to take multiple urine, saliva or serum samples. Hair grows approximately 1 centimeter per month, and hair analysis accurately reflects exposure to drug abuse and environmental toxins. Here we describe a new assay for measurement of cortisol in hair, and determined a reference range for non-obese subjects.

Methods: For measurement of cortisol in hair we modified an immunoassay originally developed for measuring cortisol in saliva. We compared hair samples obtained from various parts of the head, and assessed the effect of hair dying. We analyzed hair samples from non-obese subjects, in whom we also obtained urine, saliva and blood samples for cortisol measurements.

Results: The mean extraction recovery for hair cortisol standards of 100 ng/ml, 50 ng/ml and 2 ng/ml (n=6) was 87.9%, 88.9% and 87.4%, respectively. Hair cortisol levels were not affected by hair color or by dying hair samples after they were obtained. Cortisol levels were decreased in hair that was artificially colored before taking the sample. The coefficient of variation was high for cortisol levels in hair from different sections of the head (30.5 %), but was smaller when comparing between hair samples obtained from the vertex posterior (15.6%). The reference range for cortisol in hair was 17.7-153.2 pg/mg of hair (median 46.1 pg/mg). Hair cortisol levels correlated significantly with cortisol in 24-hour urine (r=0.33; P=0.041).

Conclusion: The correlation of hair cortisol with 24-hour urine cortisol supports its relevance as biomarker for long-term exposure.
Serum IgA1 from IgA nephropathy patients induces apoptosis in podocytes through direct and indirect pathways
Authors: Wang Cheng , Peng Hui , Tang Hua , Liu Xun , Chen Zhujiang , Lou Tanqi , Yu Xueqing
Published: May 2017

Purpose: To investigate apoptosis of podocytes induced by IgA1 isolated from IgA nephropathy (IgAN) patients through direct and indirect pathways.

Methods: Jacalin affinity chromatography and Sephacryl S-200 molecular sieve chromatography were used to isolate IgA1 from blood of IgAN patients made as aggregated IgA1 (aIgA1). Podocytes were incubated with aIgA1 or special treated medium from mesangial cells after co-incubation with aIgA1 from IgAN patients. Apoptosis of podocytes was assessed by TUNEL staining and flow cytometry. Real-time PCR was used to detect the mRNA expression of Bcl-2, Bax, Fas and Fas-L.

Results: AIgA1 from IgAN patients induced more apoptosis of podocytes by both time and concentration-dependent patterns than control (30.5±5.4% vs 20.5±4.5, respectively, P < 0.05). The percentage of apoptotic podocytes exposed to treated medium was higher than control (28.5±5.9 % vs 20.5±4.5%, respectively, P < 0.05). The level of normalized Fas mRNA expression in podocytes exposed to aIgA1 was 2.4-fold higher than control (P < 0.05), while the level in podocytes exposed with treated medium was 1.89-fold higher than control (P < 0.05), and the level of normalized Bcl-2 mRNA expression in this group was 72% lower than control (P < 0.05)

Conclusion: IgA1 from IgAN patients may induce apoptosis of podocytes through direct and indirect pathways. IgA1 may accelerate progression of IgAN by inducing apoptosis of podocytes.
Evaluation of in vivo and in vitro Biological Activity of a Vibrio Cholerae 01 Hemolysin
Authors: Arellano Galindo Jose , Rodriquez Angeles Maria Guadalupe , Guadarrama Norma Valazquez , Esteban Enrique Santos , Cerezo Silvia Giono
Published: May 2017

Purpose: To evaluate the hemolysin effect by ileal loop model produced by Vibrio cholerae O1 strains, compared with the cellular lysis or cytotoxic activity (CA) observed in cell culture.

Method: We studied nine V. cholerae O1 strains, obtained during the Mexican outbreak of cholera (1990-1993), which had CA in Vero and CHO cells. Hemolysin was monitored with the hemolysis test. Titers of CA were calculated by CD50, and the association between CA and cholera toxin (CT) production was discarded by means of neutralization tests using an anti-CT polyclonal antibody. The CT production was measured with ELISA test. The LAL assay was performed in order to study relationships between the CA and bacterial lipopolysaccharide. Strains with CA were evaluated in rabbit and rat ileal loop models; hemorrhagic fluid was also measured. Tissues from ileal loop were included in paraffin to detect intestinal epithelial damage.

Results: The hemolysin CA was not neutralized with the anti-CT polyclonal antibody. However, the associated factor of CA was heat labile. CA in cell cultures was not related to the bacterial lipopolysaccharide. The ileal loop test exhibited the presence of hemorrhagic tissue with inflammation.

Conclusion: The V. cholerae O1 strains isolated were able to secrete hemolysin which, in turn, caused CA in cell cultures and produced the hemorrhagic and inflammatory effects observed in the ileal loop of rabbit and rat models.
Impact of β-blocker treatment and nutritional status on glycemic response during exercise in patients with type 2 diabetes
Authors: Ferland Annie , Brassard Patrice , Croteau Sara , Lemieux Simone , Bergeron Jean , Lacroix Stephanie , Fournier Lison , Poirier Paul
Published: May 2017

Purpose: Most individuals with type 2 diabetes are affected by hypertension and thus have higher risk of cardiac complications. In addition to behavioural modifications, such as healthy food choices and regular physical activity, β-blocker treatment may be considered to reduce morbidity and mortality, especially after a cardiovascular event. However, this medication is generally associated with a deleterious impact on glucose metabolism. The objective of the study was to assess the impact of β-blocker treatment on glucose response during exercise in patients with type 2 diabetes, free of cardiovascular complications.

Methods: Ten sedentary men, treated with diet and/or hypoglycemic agents have performed four exercise sessions at 60% of their ￼O2peak, in the fasted state or 2 hours following a standardized breakfast, with and without β-blockers (atenolol 100 mg id for five consecutive days). Blood samples were drawn during the resting period, at 15-min intervals during the exercise session and during the recovery period.

Results: A reduction of blood glucose levels was observed following the exercise session in the postprandial state (48% and 44% reduction with and without β-blockers respectively; P < 0.001). One hour of exercise performed in the fasted state had a minimal impact on glucose and insulin levels, whether with or without β-blockers. β-blocker treatment was not associated with increased baseline blood glucose or insulin levels in the fasted or the postprandial situation.

Conclusion: Dietary status has a more important impact on plasma glucose and insulin modulation than short-term use of β-blockers.
Magnesium loss in magnesium deficient subjects with and without physical exercise during prolonged hypokinesia
Authors: Tsiamis Costas B. , Kakuris Kostas K. , Deogenov Viktor A. , Yerullis Kosmas B.
Published: May 2017

Objective: To show the effect of hypokinesia (HK; diminished movement) on magnesium (Mg2+) loss in Mg2+ deficient subjects and the effect of physical exercise and on Mg2+ deficiency with and without physical exercise: Mg2+ balance, serum Mg2+ concentration and Mg2+ loss were measured.

Methods: Studies were conducted on 30 healthy male volunteers during a pre-experimental period of 30 days and an experimental period of 364 days. They were divided equally into three-groups: unrestricted active control subjects (UACS), continuous hypokinetic subjects (CHKS) and periodic hypokinetic subjects (PHKS). The UACS group ran average distances of 9.3 ± 1.2 km.day-l; the CHKS group walked average distances of 0.9 ± 0.2 km.day-l; and the PHKS group walked and ran average distances of 0.9 ± 0.2 km and 9.3 ± 1.2 km.day-l for 5-and 2-days per week, respectively.

Results: Mg2+ deficiency, serum Mg2+ level, fecal and urine Mg2+ loss increased (P < 0.05), in the PHKS and CHKS groups compared with their pre-experimental values and the values in the UACS group. However, serum Mg2+ concentration, urine and fecal Mg2+ loss and Mg2+ deficiency increased more (P < 0.05) in the PHKS group than in the CHKS group.

Conclusions: Mg2+ deficiency is more evident with than without physical exercise and Mg2+ loss is exacerbated more with higher than lower Mg2+ deficiency. This indicates that Mg2+ deficiency with and without physical exercise and Mg2+ loss with higher and lower Mg2+ deficiency is due to inability of the body to use Mg2+ and more so when physically healthy subjects are submitted to prolonged periodic than continuous hypokinesia.
Epileptic EEG discharges and short non-convulsive crisis: Influence on cognitive and psychobehavioural functions in youths
Authors: Domizio Sergio , Conte Emanuela , Puglielli Cristina , Domizio Roberto , Verrotti Alberto , Pollice Rocco , Casacchia Massimo , Conti Chiara M. , Roncone Rita , Sabatino Giovanni , Doyle Robert , Barbacane Renato C. , Conti Pio , Sabatino Giuseppe
Published: May 2017

Purpose: Learning and behavioural difficulties often occur in benign childhood epilepsy. In recent years, several electroencephalogram (EEG) characteristics have been related to the occurrence of learning and behavioral problems. We determined if the cognitive characteristics of epileptic children depend exclusively on illness factors, or if epileptic electroencephalogram discharges during the crisis contribute to these changes.

Methods: We studied a randomly selected group of 150 youths with short non-convulsive crises, who completed cognitive testing and electroencephalographic studies. The inclusion criteria were: undefined crisis, variations in cognitive function and/or frequent epileptiform discharges on the electroencephalogram.

Results: Previous research indicates that the type of epilepsy and the patient’s educational level can influence cognitive functioning. The electroencephalographic epileptic discharges during the crisis has been found to influence cognitive transitory functions such as vigilance or swiftness of mental functions. The type of epilepsy is correlated statistically with impairment of learning ability tests: reading (F, 5.487, P = 0.005) and mathematics (F, 3.007, P ≤ 0.05). In addition, 40% of the epileptic patients had behavioural disordered versus 16% for the control group (P < 0.02).
Multidisciplinarity, interdisciplinarity, and transdisciplinarity in health research, services, education and policy: 3. Discipline, inter-discipline distance, and selection of discipline
Authors: Choi Bernard C. K. , P. Anita W.
Published: May 2017

Background/Purpose. Multiple disciplinary efforts are increasingly encouraged in health research, services, education and policy. This paper is the third in a series. The first discussed the definitions, objectives, and evidence of effectiveness of multiple disciplinary teamwork. The second examined the promoters, barriers, and ways to enhance such teamwork. This paper addresses the questions of discipline, inter-discipline distance, and where to look for multiple disciplinary collaboration.

Methods. This paper proposes a conceptual framework of the knowledge universe, based on a review of a number of key papers on the Global Brain. These key papers were identified during a literature review on multiple disciplinary teamwork, using Google and MEDLINE (1982-2007) searches.

Results. A discipline is held together by a shared epistemology. In general, disciplines that are more disparate from one another epistemologically are more likely to achieve new insight for a complex problem. The proposed conceptual framework of the knowledge universe consists of several knowledge subsystems, each containing a number of disciplines. The inter-discipline distance can guide us to select appropriate disciplines for a multiple disciplinary team.

Conclusion. If multiple disciplinarity is called for, the proposed view of the knowledge universe as a series of knowledge subsystems and disciplines, and the place of health sciences in the knowledge universe, will help researchers, practitioners, and policy makers to identify disciplines for multiple disciplinary efforts.
Human research review committee requirements in medical journals
Authors: Freeman Scott R. , Lundahl Kristy , Schilling Lisa M. , Jensen J. Daniel , Dellavalle Robert P.
Published: May 2017

Purpose: Independent ethical review committees safeguard participants in human research. The purpose of this study was to describe the current ethical guidelines for human research requirements in the Instructions to Authors of the English language medical journals previously studied in 1995.

Methods: The instructions to authors of English language medicine journals from the Abridged Index Medicus were searched for any policies regarding guidance on the ethical treatment of human subjects in research.

Results: More medical journals require independent ethics committee approval of human research now [84/101 (83%)] than 10 years ago [48/102 (47%) (P < 0.001)], and most journals continue to require that this disclosure appear in the manuscript [71/84 (85%) vs. then 37/48 (77%) (P=0.29)]. Fewer medical journal instructions to authors provide no ethical guidelines for human research now [8/101 (8%)] than 10 years ago [25/102 (24%) (P < 0.001)]. No journal required submission of the study approval letter or of the approved protocol.

Conclusions: Although medicine journals increasingly require disclosure statements of independent ethics committee approval for human research, they fail to verify such approval beyond taking authors for their word.
Microarray expression profiling of human dental pulp from single subject
Authors: Tete Stefano , Mastrangelo Filiberto , Scioletti Anna Paola , Tranasi Michelangelo , Raicu Florina , Paolantonio Michele , Stuppia Liborio , Vinci Raffaele , Gherlone Enrico , Ciampoli Cristian , Sberna Maria Teresa , Conti Pio
Published: May 2017

Introduction: Microarray is a recently developed simultaneous analysis of expression patterns of thousand of genes. The aim of this research was to evaluate the expression profile of human healthy dental pulp in order to find the presence of genes activated and encoding for proteins involved in the physiological process of human dental pulp. We report data obtained by analyzing expression profiles of human tooth pulp from single subjects, using an approach based on the amplification of the total RNA.

Methods: Experiments were performed on a high-density array able to analyse about 21,000 oligonucleotide sequences of about 70 bases in duplicate, using an approach based on the amplification of the total RNA from the pulp of a single tooth. Obtained data were analyzed using the S.A.M. system (Significance Analysis of Microarray) and genes were merged according to their molecular functions and biological process by the Onto-Express software.

Results: The microarray analysis revealed 362 genes with specific pulp expression. Genes showing significant high expression were classified in genes involved in tooth development, protoncogenes, genes of collagen, DNAse, Metallopeptidases and Growth factors.

Conclusion: We report a microarray analysis, carried out by extraction of total RNA from specimens of healthy human dental pulp tissue. This approach represents a powerful tool in the study of human normal and pathological pulp, allowing minimization of the genetic variability due to the pooling of samples from different individuals.
Chronic clofibrate administration prevents salineinduced endothelial dysfunction and oxidative stress in young Sprague-Dawley rats
Authors: Sankaralingam Sowndramalingam , Desai Kaushik M , Wilson Thomas W
Published: May 2017

Purpose: High salt intake causes hypertension and endothelial dysfunction in young Sprague-Dawley rats. Clofibrate (clof) prevents this salt induced hypertension. We asked whether clof can prevent salt-induced endothelial dysfunction, and if so, its mechanism. We also questioned whether high salt intake can induce endothelial dysfunction without hypertension in older animals.

Methods: Young (Y, 5 weeks) and old (O, 53 weeks) male Sprague-Dawley rats were given either vehicle (Con, 20 mM Na2CO3) or 0.9% NaCl (Sal) to drink for three weeks. Some young rats received clof (80 mg/d) in their drinking fluid. After three weeks, we measured mean arterial pressure (MAP), endothelial function, by comparing hypotensive responses to acetylcholine (ACh, endothelium dependent) and sodium nitroprusside (SNP, endothelium independent), plasma total nitrite+nitrate levels (PNOx), by the Griess reaction, and aortic superoxide production by lucigenin chemiluminescence.

Results: Carotid artery MAP did not change in O. Sal-Y developed hypertension: 133±3 vs. 114±2 mmHg, P < 0.001, which was prevented by clof: 105±2 mmHg. ACh induced a similar dose dependent hypotensive response in Con-O and Sal-O that was inhibited by L-NAME (100mg/kg i.v.). Responses to ACh were blunted in Sal-Y but not in Con-Y. Further, L-NAME inhibited ACh responses only in Con-Y. The response to SNP was similar in all animals. Importantly, the ACh-induced hypotensive response was potentiated in clof+Sal-Y, an effect which was attenuated by blocking calcium-activated potassium channels (KCa) with a combination of apamin (50 ug/kg i.v.) + charybdotoxin (50 ug/kg i.v.), but not by L-NAME. PNOx was reduced in Sal-Y compared to Con-Y (2.09±0.26 vs. 4.8±0.35 µM, P < 0.001), but not in Sal-O. Aortic superoxide production was higher (P < 0.001) in Sal-Y (2388±40 milliunits/mg/min) than Sal-O (1107±159 milliunits/mg/min), but was reduced by clof (1378±64 milliunits/mg/min; P < 0.001).

Conclusions: High salt intake increases oxidative stress in young animals, leading to impaired nitric oxide activity and endothelial dysfunction. Clofibrate prevents endothelial dysfunction partly through reduced O2?- formation but mainly via selective activation of KCa channels. Older animals are resistant to both salt induced hypertension and oxidative stress.
cAMP and cGMP in nasal mucus: relationships to taste and smell dysfunction, gender and age
Authors: Henkin R I , Velicu I
Published: May 2017

Purpose: To evaluate the presence and concentration of cAMP and cGMP in human nasal mucus in normal volunteers, to relate these findings to age and gender, and to compare normal levels with those in patients with taste and smell dysfunction.

Methods: Nasal mucus was collected over one to four days in 66 normal subjects and 203 patients with smell loss (hyposmia). Samples were centrifuged at 20,000 rpm, the supernatant removed and analyzed for cAMP and cGMP by using a 96 plate technique with a specific spectrophotometric colorimetric ELISA assay.

Results: Both cAMP and cGMP were present in human nasal mucus with both cAMP and cGMP significantly higher in normal women than in normal men [men vs. women; cAMP, 0.23±0.002 vs. 0.34±0.05 (P < 0.05); cGMP, 0.28±0.03 vs. 0.63±0.12 (P < 0.01)]. Both cAMP and cGMP changed with age; both moieties increased in a U shaped, parabolic pattern reaching a peak at age 41-50 with cAMP diminishing thereafter and then increasing to its highest level over age 70. Both cAMP and cGMP were lower in patients with taste and smell dysfunction than in normal subjects [normals vs. patients; cAMP, 0.31±0.05 vs. 0.15±0.02 (P < 0.01); cGMP, 0.56±0.07 vs. 0.025±0.02 (P < 0.001)] suggesting a relationship to olfactory pathology.

Conclusions: This is the first definitive study to demonstrate the presence of these cyclic nucleotides in nasal mucus and the first to reveal decreased levels in patients with impaired taste and smell function. Since olfactory receptor sensitivity decreases with age increased nasal mucus cAMP over age 70 may appear incongruous but suggests one role of cAMP in olfactory function may relate to feedback mechanism(s) whereby its increase over age 70 yr reflects a physiological attempt to enhance diminishing lfactory function through growth and development of olfactory receptor activity.