Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon

<p>Abstract</p> <p>Background</p> <p>Alpha interferon in combination with ribavirin is the standard therapy for hepatitis C virus infection. Unfortunately, a significant number of patients fail to eradicate their infection with this regimen. The mechanisms of IFN-resistance are unclear. The aim of this study was to determine the contribution of host cell factors to the mechanisms of interferon resistance using replicon cell lines.</p> <p>Results</p> <p>HCV replicons with high and low activation of the IFN-promoter were cultured for a prolonged period of time in the presence of interferon-alpha (IFN-alpha2b). Stable replicon cell lines with resistant phenotype were isolated and characterized by their ability to continue viral replication in the presence of IFN-alpha. Interferon resistant cell colonies developed only in replicons having lower activation of the IFN promoter and no resistant colonies arose from replicons that exhibit higher activation of the IFN promoter. Individual cell clones were isolated and nine IFN resistant cell lines were established. HCV RNA and protein levels in these cells were not altered by IFN- alpha2b. Reduced signaling and IFN-resistant phenotype was found in all Huh-7 cell lines even after eliminating HCV, suggesting that cellular factors are involved. Resistant phenotype in the replicons is not due to lack of interferon receptor expression. All the cell lines show defect in the JAK-STAT signaling and phosphorylation of STAT 1 and STAT 2 proteins were strongly inhibited due to reduced expression of Tyk2 and Jak-1 protein.</p> <p>Conclusion</p> <p>This in vitro study provides evidence that altered expression of the Jak-Stat signaling proteins can cause IFN resistance using HCV replicon cell clones.</p

Measured energy content of frequently purchased restaurant meals : multi-country cross sectional study

Funding: This work was supported in part by the US Department of Agriculture under agreement no. 58-1950-4-003 with Tufts University and a Tufts University Provost award to SBR. The study had additional funding in Brazil from FAPESP grants 2013/18520-0 and 2013/14489-1 to VS; in China from the National Science Foundation of China grant No 91431102 to JRS and International Partnership Program of Chinese Academy of Sciences grant No GJHZ1660 to JRS; in Finland from internal funding by the University of Eastern Finland to JP; in Ghana from the University of Georgia Global Research Collaborative Grant Program to AKA. The views expressed are those of the authors. The sponsors had no role in the design, undertaking, or reporting of the study.Peer reviewedPublisher PD

Benzodiazepine use among adults residing in the urban settlements of Karachi, Pakistan: A cross sectional study

<p>Abstract</p> <p>Background</p> <p>There are hardly any studies carried out in Pakistan on the usage of benzodiazepines at the level of community. This research was aimed to determine the frequency of benzodiazepine use, along with its associations with socio-demographic and clinical characteristics among community dwelling adults, residing in two urban settlements of Karachi, Pakistan.</p> <p>Methods</p> <p>We performed a cross sectional study from August 2008 to December 2009, in 2 areas of Karachi, namely Garden and Sultanabad. We followed the systematic sampling strategy to randomly select the households, with an adult of either sex and of age 18 years or more. Data collection was carried out through interview, using a pre-tested questionnaire, with items on socio-demographic position, medical history and benzodiazepine use. Student's t-test and χ²test was employed to determine the associations between socio-demographic and clinical characteristics, and their relationship with benzodiazepine use was determined using applied logistic regression.</p> <p>Results</p> <p>The overall percentage of benzodiazepine consumption was estimated to be 14%. There were significantly more benzodiazepine users in the peri-urban Sultanabad community to the urban community of Garden (p-value = 0.001). The mean age (± SD) for users was 51.3 (± 15.6) years compared to 37.1 (± 14.4) years among non-users. Bromazepam was the most widely used benzodiazepine (29%); followed by diazepam, with a median duration on primary use being 144 weeks (IQR = 48-240). The adjusted logistic regression model revealed that increasing age, location, female sex, unemployment and psychiatric consultation were associated with increased likelihood of benzodiazepine use.</p> <p>Conclusion</p> <p>We believe the unregulated over-the-counter sales of benzodiazepines and social conditions might be playing a role in this high consumption of benzodiazepines in the community.</p

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P &lt; 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely

Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon-4

<p><b>Copyright information:</b></p><p>Taken from "Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon"</p><p>http://www.virologyj.com/content/4/1/89</p><p>Virology Journal 2007;4():89-89.</p><p>Published online 18 Sep 2007</p><p>PMCID:PMC2075494.</p><p></p>stant Huh-7 cell lines. Cells were transfected with IRES-GFP plasmid and treated with 1000 IU/ml interferon alpha. After 24 hours, GFP expression was recorded. Cured cells prepared from resistant clones unable to activate interferon signaling and no inhibition of HCV IRES was seen

Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon-3

<p><b>Copyright information:</b></p><p>Taken from "Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon"</p><p>http://www.virologyj.com/content/4/1/89</p><p>Virology Journal 2007;4():89-89.</p><p>Published online 18 Sep 2007</p><p>PMCID:PMC2075494.</p><p></p>n of IFN promoter in the replicon cell lines was determined in the presence or absence of IFN-α2b (1000 IU/ml) after 24 hours. All nine interferon resistant cell lines have lower activation of IFN-promoter as compared to interferon sensitive cell lines

Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon-8

<p><b>Copyright information:</b></p><p>Taken from "Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon"</p><p>http://www.virologyj.com/content/4/1/89</p><p>Virology Journal 2007;4():89-89.</p><p>Published online 18 Sep 2007</p><p>PMCID:PMC2075494.</p><p></p>eplicon cell lines (9/13, 5/15 and KR) with high ISRE promoter activation were treated with or without interferon alpha 2b (1000 IU/ml) for 4 weeks in a growth medium containing G-418 (1 mg/ml). The ability of replicon cells to develop G-418 resistant cell colonies was examined. Cell colonies were developed only in low ISRE replicons. Three individual colonies from each dish were picked up and nine stable cell lines were generated

Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon-7

<p><b>Copyright information:</b></p><p>Taken from "Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon"</p><p>http://www.virologyj.com/content/4/1/89</p><p>Virology Journal 2007;4():89-89.</p><p>Published online 18 Sep 2007</p><p>PMCID:PMC2075494.</p><p></p>sferase downstream of the HCV IRES. The second cistern in the same RNA contains EMCV-IRES sequences for efficient translation of HCV non-structural proteins and this chimeric RNA terminates with the HCV 3'UTR. This sub-genomic clone has adaptive mutation S1179I that allows high level replication of RNA in Huh-7 cells and cell colonies develop that are resistant to neomycin. (B) pISRE-Luc reporter plasmid construct containing firefly luciferase reporter gene used to measure IFN-promoter activation in replicon cell lines. This construct has four copies of ISRE sequences positioned up stream of Herpes simplex virus thymidine kinase promoter TATA box, which drives the expression of firefly luciferase. (C) Represents monocistronic reporter constructs that express green fluorescent protein translated by HCV IRES by using a T7 inducible system

Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon-5

<p><b>Copyright information:</b></p><p>Taken from "Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon"</p><p>http://www.virologyj.com/content/4/1/89</p><p>Virology Journal 2007;4():89-89.</p><p>Published online 18 Sep 2007</p><p>PMCID:PMC2075494.</p><p></p>7°C for one hour. Non-specific binding was measured in presence of 100-fold excess of unlabeled IFN-α2b for each dilution. Cells were washed twice in PBS. Radioactivity (cpm) of each cell pellet was measured. The amount of interferon specifically bound was determined by subtracting non-specific binding from total binding. Specific binding of I-IFN-α2b to three groups of resistant cell lines and one sensitive cell line are shown (A-D)

Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon-1

<p><b>Copyright information:</b></p><p>Taken from "Reduced expression of Jak-1 and Tyk-2 proteins leads to interferon resistance in Hepatitis C virus replicon"</p><p>http://www.virologyj.com/content/4/1/89</p><p>Virology Journal 2007;4():89-89.</p><p>Published online 18 Sep 2007</p><p>PMCID:PMC2075494.</p><p></p>eplicon cell lines (9/13, 5/15 and KR) with high ISRE promoter activation were treated with or without interferon alpha 2b (1000 IU/ml) for 4 weeks in a growth medium containing G-418 (1 mg/ml). The ability of replicon cells to develop G-418 resistant cell colonies was examined. Cell colonies were developed only in low ISRE replicons. Three individual colonies from each dish were picked up and nine stable cell lines were generated