Assessment of treatment options for patients with hepatitis C virus recombinant form 2k/1b.

AIM: Hepatitis C virus (HCV) intergenotype recombinant form (RF) 2k/1b has been actively circulating in HCV-infected patients, and the prevalence of this RF virus in the Republic of Georgia is one of the highest reported worldwide. The aim of this study was to define the optimal treatment regimen for patients with RF_2k/1b. METHODS: We analyzed the data of 2735 patients who started treatment at the Medical Center Mrcheveli within Georgia's hepatitis C elimination program from May 2015 through December 2019. The patients were treated with sofosbuvir (SOF)-based regimens. For identification of RF_2k/1b variants, refinement of standard (INNO-LiPA) genotyping results for all patient samples assigned the unspecific HCV genotypes (GT) 2a/2c was carried out by sequencing of core and non-structural protein 5B genes. RESULTS: Overall, 444 patients, representing 66% of GT2 and 16% of the total samples, were RF_2k/1b. Treatment of patients with RF_2k/1b with SOF/ledipasvir and SOF/velpatasvir was highly effective and viral cure rates did not differ among genotypes treated with the same regimen: RF_2k/1b, 99% (343/346); GT1, 99% (876/885); GT2, 96% (156/162); and GT3, 99% (545/552). A separate comparison analysis of sustained virologic response rate, treated with SOF plus ribavirin, showed significantly higher sustained virologic response (96%) in patients with confirmed GT2 (by sequencing) compared to unspecified GT2 (by INNO-LiPA) (79%) (P < 0.05). CONCLUSION: Sofosbuvir-based regimens are highly effective for treatment of RF 2k/1b patients, and with availability of new pan-genotypic direct-acting antivirals, genotyping to identify RF 2k/1b patients might not be necessary.

Identification of hepatitis C virus 2k/1b intergenotypic recombinants in Georgia.

BACKGROUND AND AIMS: This study aimed to evaluate the prevalence of the hepatitis C virus intergenotype recombinant strain RF1_2k/1b in Georgia, confirm viral recombination by full genome sequencing, and determine a genetic relationship with previously described recombinant hepatitis C viruses. METHODS: We retrospectively analysed data from 1421 Georgian patients with chronic hepatitis C. Genotyping was performed with the INNO-LiPA VERSANT HCV Genotype 2.0 Assay. RESULTS: Virus isolates were assigned to nonspecific hepatitis C genotypes 2a/2c (n = 387) as performed by sequencing of core and NS5B genes. Subsequently, sequencing results classified the core region as genotype 2k and the NS5B region as genotype 1b for 72% (n = 280) of genotype 2 patients, corresponding to 19.7% of hepatitis C patients in Georgia. Eight samples were randomly selected for full genome sequencing which was successful in 7 of 8 samples. Analysis of the generated consensus sequences confirmed that all 7 viruses were 2k/1b recombinants, with the recombination breakpoint located within 73-77 amino acids before the NS2-NS3 junction, similar to the previously described RF1_2k/1b virus. Phylogenetic analysis revealed clustering of the Georgian 2k/1b viruses and RF1_2k/1b, suggesting that they are genetically related. CONCLUSIONS: The 19.7% prevalence of RF1_2k/1b in Georgia patients is far higher than has generally been reported to date worldwide. Identification of recombinants in low income countries with a high prevalence of HCV infection might be reasonable for choosing the most cost-effective treatment regimens.

Emergence of Hepatitis C Virus Genotype Recombinant Forms 2k/1b in Georgia.

BACKGROUND: Hepatitis C virus (HCV) evolution is thought to proceed by mutations within the six major genotypes. Studies of HCV recombinant genotypes in different parts of the world have recently been initiated. Only a few cases of recombination have been identified worldwide, predominantly in Eastern Europe and Asia. In 2011 we detected the recombinant form (RF) of a HCV genotype RF_2k/1b in Georgia. Therefore, we reviewed HCV genotyping data of 491 patients with chronic hepatitis C virus infections of our center in Tbilisi over a period of two years. METHODS: Initially all genotyping analyses were performed with the VERSANT HCV genotype assay (Siemens, LiPA). In a second analysis, parts of the core and the NS5B region were sequenced for all HCV genotypes 2a/2c. RESULTS: Approximately 2/3 of genotype 2 cases were identified as the recombinant form HCV-RF 2k/1b. Overall, this type represented 19% of all HCV patients who underwent genotyping. CONCLUSIONS: We can conclude that almost 20% of HCV infected Georgian patients are infected with HCVRF_2k/ 1b.

Iohexol plasma clearance measurement in older adults with chronic kidney disease-sampling time matters.

BACKGROUND: Accurate and precise measurement of GFR is important for patients with chronic kidney disease (CKD). Sampling time of exogenous filtration markers may have great impact on measured GFR (mGFR) results, but there is still uncertainty about optimal timing of plasma clearance measurement in patients with advanced CKD, for whom 24-h measurement is recommended. This satellite project of the Berlin Initiative Study evaluates whether 24-h iohexol plasma clearance reveals a clinically relevant difference compared with 5-h measurement in older adults. METHODS: In 104 participants with a mean age of 79 years and diagnosed CKD, we performed standard GFR measurement over 5 h (mGFR300) using iohexol plasma concentrations at 120, 180, 240 and 300 min after injection. With an additional sample at 1440 min, we assessed 24-h GFR measurement (mGFR1440). Study design was cross-sectional. Calculation of mGFR was conducted with a one compartment model using the Brochner-Mortensen equation to calculate the fast component. mGFR values were compared with estimated GFR values (MDRD, CKD-EPI, BIS1, Revised Lund-Malmö and Cockcroft-Gault). RESULTS: In all 104 subjects, mGFR1440 was lower than mGFR300 (23 ± 8 versus 29 ± 9 mL/min/1.73 m(2), mean ± SD; P < 0.001). mGFR1440 was highly correlated with mGFR300 (r = 0.9). The mean absolute difference mGFR300 - mGFR1440 was 5.9 mL/min/1.73 m(2) corresponding to a mean percentage difference of 29%. In individuals with eGFRCKD-EPI ≤ 30 mL/min/1.73 m(2), percentage difference of mGFR300 and mGFR1440 was even higher (35%). To predict mGFR1440 from mGFR300, we developed the correction formula: mGFR1440 = -2.175 + 0.871 × mGFR300 (1-fold standard error of estimate: ±2.3 mL/min/1.73 m(2)). The GFR estimating equation with the best accuracy and precision compared with mGFR300 and mGFR1440 was the Revised Lund Malmö. CONCLUSIONS: In elderly CKD patients, measurement of iohexol clearance up to 5 h leads to a clinically relevant overestimation of GFR compared with 24-h measurement. In clinical care, this effect should be bore in mind especially for patients with considerably reduced GFR levels. A new correction formula has been developed to predict mGFR1440 from mGFR300. For accurate GFR estimates in elderly CKD patients, we recommend the Revised Lund Malmö equation.

Laser microdissection of paraffin embedded tissue as a tool to estimate the sialylation status of selected cell populations.

In vertebrates, sialic acids occur at the terminal end of glycans mediating numerous biological processes like cell differentiation or tumor metastasis. Consequently, the cellular sialylation status under healthy and pathological conditions is of high interest. Existing analytical strategies to determine sialylation patterns are mostly applied to tissue samples consisting of a mixture of different cell types. Alterations in the sialylation status in a distinct area of tissues or in a specific cell population may, therefore, be easily overlooked. Likewise, estimated variations in sialylation in tissue homogenates might be simply the result of a changed cell composition. To overcome these limitations, we employed laser microdissection to isolate defined cell types or functional subunits and cell populations of paraffin embedded specimens which represent the most abundant supply of human tissue associated with clinical records. For qualitative and quantitative estimation of the sialylation status, sialic acids were released, fluorescently labeled, and analyzed by an online high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) system. As a proof of principle, this strategy was successfully applied to characterize the sialylation of the apical region of epididymal epithelial cells. Furthermore, it was possible to detect an impaired sialylation during kidney maturation in a transgenic mouse model, which was restricted to glomeruli, whereas no differences in sialylation were observed when whole kidney homogenates were used. Thus, starting from paraffin embedded tissue samples, the outlined approach offers a sensitive method to detect and quantify sialic acids on defined cell populations, which may be useful to explore novel sialic acid dependent roles during physiological and pathological processes.

Author Correction: Beta Trace Protein does not outperform Creatinine and Cystatin C in estimating Glomerular Filtration Rate in Older Adults.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.

Clinical effectiveness of balloon dacryocystoplasty in circumscribed obstructions of the nasolacrimal duct.

PURPOSE: Balloon dacryocystoplasty (DCP) is known to have limited clinical success rates. In a previous publication, we tried to define subgroups that could benefit from interventional treatment of tear duct stenoses. The purpose of this study was to evaluate the clinical effectiveness of DCP performed with limited indication confined to patients with circumscribed stenosis or distal occlusion of the nasolacrimal duct (NLD). PATIENTS AND METHODS: Twenty-nine patients with severe epiphora due to dacryocystographically proven postsaccal obstruction of the lacrimal draining system were treated by means of DCP and were available for a telephone interview after a median follow-up period of 40 months (5-75 months). A standardized questionnaire covered the individual history of epiphora before and after interventional treatment. All patients had circumscribed stenoses of the lower lacrimal sac or the NLD or presented with short-distance occlusions of the distal NLD. Patients with canalicular, high saccal or diffuse lesions as well as cases with active dacryocystitis, suspicion of dacryocystolithiasis or posttraumatic stenosis were excluded from DCP. Failures or recurrences with no major improvement compared with the initial status were taken as a study endpoint. RESULTS: We dilated 21 partial and 8 complete obstructions and post-DCP control dacryocystograms showed a widening of the ductal lumen or improvement of flow. In 25 out of 29 patients, regression of clinical symptoms occurred during the first week after treatment, 4 cases remained unchanged. Ten out of 25 patients with initial improvement reported recurrence of severe epiphora after a median period of 5 months. Five patients of all treated patients (n = 29) received additional operative dacryocystorhinostomy (DCR) after failure of DCP or due to severe recurrences. One patient received DCR during the 1st week after DCP (n = 1). Four out of 25 patients with initial improvement underwent DCR. Overall, 15 of the 29 treated patients had durable improvement of epiphora by DCP alone. CONCLUSION: Even patients with circumscribed obstructions of the NLD and exclusion of factors potentially associated with poor outcome of tear duct stenosis after DCP balloon dilatation showed a limited clinical success and high recurrence rate. The main argument to continue DCP as first or second line treatment in selected patients with duct obstructions is its lack of invasiveness. Even patients with increased risk of general anesthesia can be treated and approximately half of the operations may be avoided.

Beta Trace Protein does not outperform Creatinine and Cystatin C in estimating Glomerular Filtration Rate in Older Adults.

Despite intense research the optimal endogenous biomarker for glomerular filtration rate (GFR) estimation has not been identified yet. We analyzed if ß-trace protein (BTP) improved GFR estimation in elderly. 566 participants aged 70+ from the population-based Berlin Initiative Study were included in a cross-sectional validation study. BTP, standardized creatinine and cystatin C were measured in participants with iohexol clearance measurement as gold standard method for measured GFR (mGFR). In a double logarithmic linear model prediction of mGFR by BTP was assessed. Analyses with BTP only and combined with creatinine and cystatin C were performed. Additionally, performance of GFR estimating equations was compared to mGFR. We found that the combination of all three biomarkers showed the best prediction of mGFR (r2 = 0.83), whereat the combination of creatinine and cystatin C provided only minimally diverging results (r2 = 0.82). Single usage of BTP showed worst prediction (r2 = 0.67) within models with only one biomarker. Subgroup analyses (arterial hypertension, diabetes, body mass index ≤23 and >30) demonstrated a slight additional benefit of including BTP into the prediction model for diabetic, hypertensive and lean patients. Among BTP-containing GFR equations the Inker BTP-based equation showed superior performance. Especially the use of cystatin C renders the addition of BTP unnecessary.

Cystatin C standardization decreases assay variation and improves assessment of glomerular filtration rate.

BACKGROUND: Cystatin C is increasingly used in glomerular filtration rate (GFR) estimation equations. The dependence of cystatin C results upon the analytical method has been a major source of controversy. METHODS: Cystatin C was measured with non-standardized turbidimetric Roche Generation 1 and standardized nephelometric Siemens assays in 3666 and additionally with standardized Roche Generation 2 and Siemens in 567 blood samples of the Berlin Initiative Study. Cystatin C-based GFR was assessed with CKD-EPIcys (Chronic Kidney Disease Epidemiology) and CAPA (Caucasian, Asian, Pediatric, Adult) equations and the impact of the assays on GFR estimation was determined. Equation performance compared to measured GFR was evaluated. RESULTS: Concordance of Roche Gen2 and Siemens was high with median difference of 0.003 ± 0.13 mg/L (limits of agreement: -0.12 to 0.12) and Passing Bablok correlation was essentially perfect. Roche Gen1 assay showed worse concordance with Siemens: median difference was 0.08 ± 0.13 mg/L (limits of agreement: -0.18 to 0.34) and correlation was inferior. Mean difference (± SD) of estimated GFRCKD-EPIcys was 0 ± 4 mL/min/1.73 m(2) for Gen2 and Siemens compared to -5 ± 8 with Gen1. Performance of GFR estimating equations was not influenced by the choice of Siemens or Gen2 assays. CONCLUSIONS: Standardization of Roche Gen2 assay improved accuracy of cystatin C measurement compared to Siemens. It suggests only negligible method bias and results in equal performance of both assays when estimating GFR indicating that successful calibration has led to major progress in cystatin C analysis.

Association between cerebral ischemia and cytotoxin-associated gene-A-bearing strains of Helicobacter pylori.

BACKGROUND AND PURPOSE: Studies on Helicobacter pylori infection and risk of ischemic stroke yielded variable results. Infection with more virulent H. pylori strains, such as cytotoxin-associated gene-A (CagA)-bearing strains, may be of particular relevance for ischemic diseases. We investigated whether H. pylori and CagA seropositivity are independent risk factors for cerebral ischemia or its etiologic subtypes. METHODS: We determined IgG antibodies against H. pylori and CagA protein (enzyme immunoassays) in 190 patients with acute cerebral ischemia and in 229 age- and sex-matched control subjects selected randomly from the general population. RESULTS: CagA seropositivity was more common in patients (114/190; 60.0%) than in control subjects (99/229; 43.2%; odds ratio, 1.97; 95% CI, 1.33 to 2.91; P<0.001). This result remained significant after adjustment for age, sex, vascular risk factors and diseases, and childhood and adult social status (odds ratio, 1.84; 95% CI, 1.13 to 3.00; P=0.015). Subgroup analyses yielded similar results in all etiologic stroke subtypes. In contrast, H. pylori seropositivity in general was not associated with increased risk of stroke or its etiologic subtypes. CONCLUSIONS: Our results support the hypothesis of an association between infection with CagA-positive H. pylori strains and acute cerebral ischemia.

Comparison of different salivary and fecal antibodies for the diagnosis of celiac disease.

To investigate the detectability and expressiveness of salivary and fecal anti-gliadin (AGA), anti-endomysium (EMA) and anti-tissue-transglutaminase (ATA) antibodies, 127 salivary and 160 fecal samples of healthy volunteers and salivary and fecal samples of 17 patients with histologically proven and 9 patients with suggested celiac disease were investigated in this study. With all salivary parameters and fecal IgA AGA, IgM AGA, IgA EMA and IgG EMA, healthy volunteers and patients showed partially overlapping results. The most promising results in our study with higher concentrations in patients with celiac disease were obtained by fecal scIgA AGA and a combined determination of fecal IgA AGA, IgG AGA and IgM AGA. Further investigations should be performed with fecal IgA EMA and scIgA ATA based on human recombinant tissue-transglutaminase. One patient with histologically proven celiac disease had normal serological but high fecal scIgA AGA and scIgA ATA values. This patient emphasizes the importance of fecal antibody determination for the diagnosis of celiac disease, at least in patients with suggested celiac disease and negative serum antibodies.

[Outbreak of measles in Kielce--June 2001]. / Ognisko odry w Kielcach--czerwiec 2001.

Kielce area has had a very low incidence of measles. The outbreak of measles was reported in June 2001 and included a cumulative number of 12 cases, of whom 11 have been lab confirmed. Nine cases belonged to Gypsy community and were less than 16 years of age. Remaining 3 cases were medical care workers of age 29-31. None of the children involved in the outbreak have been previously vaccinated against measles.

Occurrence of renal and pulmonary syndrome in a region of northeast Germany where Tula hantavirus circulates.

Hantavirus species Tula (TULV) is carried by European common voles (Microtus spp.). Its pathogenic potential for humans is unknown. In a rural region of northeast Germany, a 43-year-old man became ill with fever, renal syndrome, and pneumonia. Typing of late acute- and convalescent-phase sera by focus reduction neutralization assay revealed the presence of neutralizing antibodies against TULV. Moreover, we detected TULV genetic material in Microtus arvalis animals that were trapped at places only a few kilometers from the home village of the patient. Phylogenetic analysis of completely sequenced genomic S segments from three virus strains grouped them within a third genetic lineage of the TULV species. This is the first case of hemorrhagic fever with renal syndrome and pulmonary involvement which can be associated with TULV infection.