In Vitro Characterization of GSK2485852, a Novel Hepatitis C Virus Polymerase Inhibitor

GSK2485852 (referred to here as GSK5852) is a hepatitis C virus (HCV) NS5B polymerase inhibitor with 50% effective concentrations (EC₅₀s) in the low nanomolar range in the genotype 1 and 2 subgenomic replicon system as well as the infectious HCV cell culture system. We have characterized the antiviral activity of GSK5852 using chimeric replicon systems with NS5B genes from additional genotypes as well as NS5B sequences from clinical isolates of patients infected with HCV of genotypes 1a and 1b. The inhibitory activity of GSK5852 remained unchanged in these intergenotypic and intragenotypic replicon systems. GSK5852 furthermore displays an excellent resistance profile and shows a <5-fold potency loss across the clinically important NS5B resistance mutations P495L, M423T, C316Y, and Y448H. Testing of a diverse mutant panel also revealed a lack of cross-resistance against known resistance mutations in other viral proteins. Data from both the newer 454 sequencing method and traditional population sequencing showed a pattern of mutations arising in the NS5B RNA-dependent RNA polymerase in replicon cells exposed to GSK5852. GSK5852 was more potent than HCV-796, an earlier inhibitor in this class, and showed greater reductions in HCV RNA during long-term treatment of replicons. GSK5852 is similar to HCV-796 in its activity against multiple genotypes, but its superior resistance profile suggests that it could be an attractive component of an all-oral regimen for treating HCV.     

Calibration of estimated biting forces in domestic canids: comparison of post-mortem and in vivo measurements

Estimates of biting forces are widely used in paleontological and comparative studies of feeding mechanics and performance, and are usually derived from lever models based on measurements made on the skull that are relevant to the mechanics of the masticatory system. Owing to assumptions and unmeasurable errors in their estimation, such values are used comparatively rather than as absolute estimates. The purpose of this paper was to provide calibration of post-mortem calculated bite force estimates by comparing them to in vivo forces derived from a sample of 20 domestic dogs ( Canis familiaris ) during muscle stimulation under general anaesthesia. Two lever models previously described in the literature were used to estimate post-mortem values, and regression analysis was also performed to derive best-fit equations against a number of morphometric measurements on the skull. The ranges of observed forces in vivo were 147–946 N at the canine, and 524–3417 N at the second molar. The lever models substantially underestimated these forces, giving mean values between 39% and 61% of the observed means. Predictability was considerably improved by removing the linear bias and deviation of the regression slope from unity with an adjustment equation. Best-fit statistical models developed on these animals performed considerably better (calculated means within 0.54% of observed means) and included easily measureable variables such as bodyweight, dimensions of the temporalis fossa and out-lever from the jaw joint to the biting tooth. These data should lead to more accurate absolute, rather than relative, estimates of biting forces for other extant and fossil canids, and other carnivorans by extrapolation.     

Blood safety & prevalence of transfussion transmissible viral infections among donors at the Red Cross Blood Bank in Gondar University Hospital

BACKGROUND: Although millions of lives are saved by blood transfusion, unsafe transfusion is also putting millions of people at risk of Transfusion Transmissible Infections (TTI). In Ethiopia the safety and magnitude of blood transfusion and TTIs is not well investigated OBJECTIVE: to investigate the magnitude of TTIs and assess TTI related risk factors among blood donors. METHODS: Cross-sectional study among blood donors at the Red Cross Society Blood Bank in Gondar University Hospital, North West Ethiopia. RESULTS: The study was conducted among 600 adult blood donors between April and July 2004. Their mean age was 28 +/- 10.4 years (range: 16-64) and 66% of them were urban dwellers. The donors included farmers (30.7%), daily laborers (22.7%) and students (20%). All the donors were first time donors and 75% of the donations were replacement donations by remunerated donors or family members. The prevalence of HIV, Hepatitis B surface antigen (HBSAg) and Hepatitis C virus (HCV) infections were 4.5% (95%CI: 3.0-6.6), 8.2% (95%CI: 6.2-10.7) and 5.8% (95%CI: 4.2-8.1), respectively. The overall discard rate of donated blood because of these viral markers was 16.3%. In univariate analysis, HCV infection was significantly associated with presence of HIV (OR: 5.36, 95%CI: 2-14.3). Experiences of traditional surgical incisions or phlebotomy were present in 93.8% of donors. While 6.5% of donors admitted a history of multiple sexual partners, none of them admitted a history of any sexually transmitted infections (STI). CONCLUSION: The prevalence of TTIs among blood donors is very high. The majorities of blood donors are replacement or paid donors with one or more of the risk factors for TTIs implying that blood transfusion is unsafe. These findings call for the urgent implementation of the national strategy for safe blood transfusion in Ethiopia. As the utilization of advanced technology and skilled-personnel-based screening of blood are not in the immediate horizon, establishing stringent selection criteria of donors and setting clinical indications for transfusion would be indispensable and cost-effective interventions to minimize the risk of TTIs to blood recipients in Ethiopia.     

Impact of common epidermal growth factor receptor and HER2 variants on receptor activity and inhibition by lapatinib

The goal of this study was to characterize the effects of non-small cell lung carcinoma (NSCLC)-associated mutations in epidermal growth factor receptor (EGFR/ErbB1) and HER2 (ErbB2) on interactions with the dual tyrosine kinase inhibitor lapatinib. Biochemical studies show that commonly observed variants of EGFR [G719C, G719S, L858R, L861Q, and Delta746-750 (del15)] are enzyme activating, increasing the tyrosine kinase V(max) and increasing the K(m)((app)) for ATP. The point mutations G719C and L861Q had minor effects on lapatinib K(i)s, whereas EGFR mutations L858R and del15 had a higher K(i) for lapatinib than wild-type EGFR. Structural analysis of wild-type EGFR-lapatinib complexes and modeling of the EGFR mutants were consistent with these data, suggesting that loss of structural flexibility and possible stabilization of the active-like conformation could interfere with lapatinib binding, particularly to the EGFR deletion mutants. Furthermore, EGFR deletion mutants were relatively resistant to lapatinib-mediated inhibition of receptor autophosphorylation in recombinant cells expressing the variants, whereas EGFR point mutations had a modest or no effect. Of note, EGFR T790M, a receptor variant found in patients with gefitinib-resistant NSCLC, was also resistant to lapatinib-mediated inhibition of receptor autophosphorylation. Two HER2 insertional variants found in NSCLC were less sensitive to lapatinib inhibition than two HER2 point mutants. The effects of lapatinib on the proliferation of human NSCLC tumor cell lines expressing wild-type or variant EGFR and HER2 cannot be explained solely on the basis of the biochemical activity or receptor autophosphorylation in recombinant cells. These data suggest that cell line genetic heterogeneity and/or multiple determinants modulate the role played by EGFR/HER2 in regulating cell proliferation.     

Diarrhoea-associated parasites in HIV-1 seropositive and sero-negative patients in a teaching hospital, Northwest Ethiopia

OBJECTIVE: to determine the prevalence and type of intestinal parasites in HIV infected and uninfected patients with diarrhea. DESIGN: A cross-sectional study was conducted at Gondar University hospital, Northwest Ethiopia, between March 2003 and October 2004. PATIENTS AND METHODS: A total 312 consecutive diarrheic patients were included in the study. Stool specimens were collected and examined for intestinal parasites following direct, formol-ether concentration and modified acid fast staining methods. RESULTS: Among the patients, 63.8% were found to be HIV seropositive. The prevalence of intestinal parasites in HIV seropositive and seronegative diarrheic patients was 30.6% and 33.6%, respectively. The most prevalent parasites were Strongyoides stercoralis (9.0%) and Entamoeba histolytica (8.3%) followed by Ascaris lumbricoides (5.4%) and Cryptosporidium species (5. 1%). CONCLUSION AND RECOMMENDATION: The prevalence of intestinal parasites in diarrheic patients was very high. Institution of appropriate intervention measures are needed to reduce morbidity in such patients.     

Constant Transmission Properties of Variant Creutzfeldt-Jakob Disease in 5 Countries

Variant Creutzfeldt-Jakob disease (vCJD) has been reported in 12 countries. We hypothesized that a common strain of agent is responsible for all vCJD cases, regardless of geographic origin. To test this hypothesis, we inoculated strain-typing panels of wild-type mice with brain material from human vCJD case-patients from France, the Netherlands, Italy, and the United States. Mice were assessed for clinical disease, neuropathologic changes, and glycoform profile; results were compared with those for 2 reference vCJD cases from the United Kingdom. Transmission to mice occurred from each sample tested, and data were similar between non-UK and UK cases, with the exception of the ranking of mean clinical incubation times of mouse lines. These findings support the hypothesis that a single strain of infectious agent is responsible for all vCJD infections. However, differences in incubation times require further subpassage in mice to establish any true differences in strain properties between cases.     

Systemic hormonal contraception initiation after abortion: A systematic review and meta-analysis

BackgroundImmediate contraceptive initiation, including start of a method before abortion completion, is a convenient option for women seeking abortion care.ObjectivesTo evaluate the effect of systemic hormonal contraception initiation on medical abortion effectiveness and the safety of hormonal contraceptive methods following abortion.Data sourcesPubMed, Popline, Cochrane Library, and Clinicaltrials.gov.Study eligibility criteriaStudies that assessed medical abortion effectiveness after systemic hormonal contraception initiation and the safety of hormonal contraception initiation after abortion.ParticipantsPregnant persons undergoing or who had recently undergone an abortion.InterventionsInitiation of systemic hormonal contraception post abortion or on the day of the first pill of the medical abortion.Study appraisal and synthesis methodsWe assessed study quality using the US Preventive Services Task Force evidence grading system. We created narrative summaries and calculated pooled relative risks when appropriate.ResultsWe identified 16 studies for inclusion, 7 randomized controlled trials, and 9 cohorts. Nine studies assessed medical abortion effectiveness with hormonal contraception initiation and generally found no decreased risk of abortion success or increased risk of additional treatment. One fair-quality study reported a small increase in ongoing pregnancy rate with immediate depot medroxyprogesterone (DMPA) compared with delayed DMPA initiation (3.6% vs 0.9%, risk difference 2.7%, 90% confidence interval 0.4–5.6). We identified no bleeding-related safety concerns following hormonal contraception initiation after medical or surgical abortion. Pooled results were too imprecise to draw firm conclusions.LimitationsIncluded studies were poor or fair quality and primarily in high-income or upper-middle-income settings.ConclusionsAbortion effectiveness did not differ between immediate vs delayed initiation of most systemic hormonal contraceptive methods after a first trimester medical abortion. However, immediate DMPA initiation did show increased ongoing pregnancy. Bleeding effects with hormonal contraception initiation postabortion appeared minimal.ImplicationsInitiating a hormonal contraceptive method after an abortion and as early as the same day as the first pill of the medical abortion is an option if contraception is desired. The slight increase in ongoing pregnancy with immediate DMPA initiation highlights the importance of information provision during contraceptive counseling.     

Levels and Trophic Transfer of Selected Pesticides in the Lake Ziway Ecosystem

The levels of 30 selected pesticides and trophic biomagnification of DDT were investigated in biota samples of the Lake Ziway in the Rift valley region, Ethiopia. Carbon source and trophic position were calculated by using ¹³C and ¹⁵N stable isotopes, individually, and trophic magnification factors (TMFs) were inferred. Only DDT and its metabolites were quantified in all samples analyzed. The most prominent metabolite was p,p?-DDE with mean concentration ranging from the 0.82–33.69 ng g^?1 lipid weight. Moreover, the ratio of DDT/DDD?+?DDE in all the biota samples was less than 1 signifying historical DDT application. Regression of log [ΣDDT] vs TL (trophic level) among all biota species showed a significant correlation, indicating that DDTs are biomagnifying along with the food web of Lake Ziway with an estimated TMF of 2.75. The concentrations of DDTs and other organochlorine pesticides found in biota from Lake Ziway were, in general, lower than studies found in previous studies carried out in the same lake.