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BACKGROUND AND AIMS: Rubus ser. Glandulosi provides a unique model of geographical parthenogenesis on a homoploid (2n = 4x) level. We aim to characterize evolutionary and phylogeographical patterns in this taxon and shed light on the geographical differentiation of apomicts and sexuals. Ultimately, we aim to evaluate the importance of phylogeography in the formation of geographical parthenogenesis. METHODS: Rubus ser. Glandulosi was sampled across its Eurasian range together with other co-occurring Rubus taxa (587 individuals in total). Double-digest restriction site-associated DNA sequencing (ddRADseq) and modelling of suitable climate were used for evolutionary inferences. KEY RESULTS: Six ancestral species were identified that contributed to the contemporary gene pool of R. ser. Glandulosi. Sexuals were introgressed from Rubus dolichocarpus and Rubus moschus in West Asia and from Rubus ulmifolius agg., Rubus canescens and Rubus incanescens in Europe, whereas apomicts were characterized by alleles of Rubus subsect. Rubus. Gene flow between sexuals and apomicts was also detected, as was occasional hybridization with other taxa. CONCLUSIONS: We hypothesize that sexuals survived the last glacial period in several large southern refugia, whereas apomicts were mostly restricted to southern France, whence they quickly recolonized Central and Western Europe. The secondary contact of sexuals and apomicts was probably the principal factor that established geographical parthenogenesis in R. ser. Glandulosi. Sexual populations are not impoverished in genetic diversity along their borderline with apomicts, and maladaptive population genetic processes probably did not shape the geographical patterns.

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Asexual organisms often differ in their geographic distributions from their sexual relatives. This phenomenon, termed geographic parthenogenesis, has long been known, but the underlying factors behind its diverse patterns have been under dispute. Particularly problematic is an association between asexuality and polyploidy in most taxa. Here, we present a new system of geographic parthenogenesis on the tetraploid level, promising new insights into this complex topic. We used flow cytometric seed screen and microsatellite genotyping to characterise the patterns of distribution of sexuals and apomicts and genotypic distributions in Rubus ser. Glandulosi across its range. Ecological modelling and local-scale vegetation and soil analyses were used to test for niche differentiation between the reproductive groups. Apomicts were detected only in North-western Europe, sexuals in the rest of the range in Europe and West Asia, with a sharp borderline stretched across Central Europe. Despite that, we found no significant differences in ecological niches. Genotypic richness distributions suggested independence of the reproductive groups and a secondary contact. We argue that unless a niche differentiation (resulting from polyploidy and/or hybridity) evolves, the main factors behind the patterns of geographic parthenogenesis in plants are phylogeographic history and neutral microevolutionary processes, such as clonal turnover.

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In a group of 211 patients with chronic hepatitis C treated with direct-acting antivirals, four experienced therapy failure. Two patients, one originally treated with dasabuvir/ombitasvir/paritaprevir/ritonavir and the other with glecaprevir/pibrentasvir, received a triple combination of sofosbuvir, velpatasvir and voxilaprevir for 12 weeks. Following the retreatment, both patients were permanently virus-free.

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BACKGROUND: Vertical hepatitis C virus (HCV) transmission and persistence of anti-HCV antibodies were retrospectively investigated since 1999 in a group of 244 children whose mothers had a history of hepatitis C. MATERIAL AND METHODS: Initial examinations performed in most children at 6 months of age included the determination of anti-HCV antibodies, HCV nucleic acid (HCV RNA), and anti-HIV antibodies, with all children being negative for HIV. Further examinations with investigation of anti-HCV and HCV RNA were performed at half-year intervals until the disappearance of anti-HCV antibodies. Vertical HCV transmission was defined by HCV RNA positivity in at least 2 venous blood samples or at least two positive anti-HCV results in a child over 3 years of age. RESULTS: Vertical HCV transmission was detected in 11 out of 244 children (4.5%). Only 2 children spontaneously cleared HCV; positive anti-HCV antibodies were last detected when they were 8 years old. Chronic hepatitis C developed in 9 children, four of whom were infected with genotype 1b, 3 children with genotype 3a, one with genotype 1a, and the last one with genotypes 1a and 4. Antiviral treatment including conventional or pegylated interferon, or ribavirin, was administered to 3 children, with sustained elimination of the virus in 2 children. Although the proportion of children with positive anti-HCV antibodies declined gradually, anti-HCV positivity was reported in 6 uninfected children at 18 months of age but in none of them at the age of 2 years. CONCLUSIONS: Vertical transmission of HCV was found in 11 out of 244 children; chronic hepatitis C was detected in 9 children; uninfected children cleared anti-HCV antibodies by 2 years of age.

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OBJECTIVE: To determine the prevalence of autoimmune parameters in patients with chronic hepatitis B and C (HBV, HCV) treated with conventional or pegylated interferon alpha (IFN) and monitor the development of autoimmune diseases in connection with this treatment. PATIENTS AND METHODS: In the years 1992-2014, autoimmune parameters were evaluated in 324 patients (271 with HCV, 53 with HBV) treated with IFN at the Department of Infectious Diseases in Ostrava. Prior to, during and after completion of IFN treatment, antinuclear antibodies (ANA), antimitochondrial antibodies (AMA), smooth muscle antibodies (SMA), anti-liver/kidney microsomal antibodies (anti-LKM-1), anti-double-stranded DNA antibodies (anti-ds-DNA), antibodies against granulocytes (ANCA), anti-deoxyribonucleoprotein antibodies (anti-DNP), anti-nucleosomes antibodies, rheumatoid factor (RF) and circulating immune complexes (CIC) were determined and clinical manifestations of autoimmune diseases were evaluated. RESULTS: At least one abnormal parameter was present in 267 of 324 patients: ANA in 140, AMA in 13, SMA in 100, RF in 118, ANCA in 11, anti-ds-DNA in 2 and anti-LKM-1 in 1 patient. Increases in CIC were observed in 150 of 227 patients, anti-DNP positivity in 39 of 239 and anti-nucleosomes were positive in none of 43 patients. At least one abnormal parameter was detected in 85 % of patients with HCV and in 89 % of patients with hepatitis B, in 81 % of patients under 40 years of age and in 84 % of older indivi-duals, 90 % of patients with cirrhosis and 80 % without cirrhosis, in 74 % of patients with treatment shorter than 30 weeks and in 87 % of patients with treatment lasting over 50 weeks. Autoimmune diseases - autoimmune hepatitis, autoimmune myositis, myopathy and diabetes - developed in 4 patients while only 3 individuals had ANA, SMA or anti-DNP positivity. CONCLUSIONS: Positivity of ANA and SMA or increased RF and CIC are often found in patients with HBV and HCV treated with IFN, but their presence does not correlate with the development of autoimmune diseases.

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OBJECTIVE: To determine the incidence of thyroid dysfunction in patients with chronic hepatitis B and C (HBV, HCV) who were treated with interferon (IFN) alpha. PATIENTS AND METHODS: In the years 1992-2013, parameters of the thyroid gland were evaluated in 304 patients (256 with HCV, 48 with HBV) who were treated with conventional or pegylated IFN at the Department of Infectious Diseases in Ostrava. Prior to, during and after completion of antiviral treatment, levels of thyroid stimulating hormone (TSH), thyroxine (T4), triiodothyronine (T3), including their free fractions fT4 and fT3, and anti-thyroid antibodies (anti-thyroglobulin, anti-microsomal fraction) were determined and clinical manifestations of thyroid dysfunction were evaluated. RESULTS: TSH changes were detected in 75 patients (25 %), of whom 68 had HCV and 7 HBV. Hypothyroidism was detected in 39 patients (34 with HCV), of whom 25 required substitute therapy which was subsequently terminated in 5 patients. Hyperthyroidism with transient suppressive therapy with carbimazole developed in 4 HCV patients. In 32 patients, TSH changes were assessed as subclinical hypothyroidism. Abnormal T3 values were found in 188 (62 %) and T4 in 49 (16 %) patients; these changes practically did not correlate with TSH changes. Autoantibodies were detected in 54 (18 %) patients of whom 30 were also found to have changes in TSH. CONCLUSIONS: In a group of 304 patients treated with IFN alpha for chronic hepatitis, thyroid disease with changes in TSH were observed in a quarter of patients; hypothyroidism clearly prevailed. Thyroid diseases developed in half of the patients with the presence of antithyroid antantibodies.

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In this work a simple technique employing oxalic and nitric acid to cow's milk samples prior to analysis by inductively coupled plasma orthogonal acceleration time-of-flight mass spectrometry (ICP-oa-TOF-MS) was introduced. After the precipitation of calcium and proteins via oxalic and nitric acid, respectively, the resulting liquid phase was aspirated with a concentric glass nebulizer for ICP-TOF-MS determination of trace elements. Precipitation of proteins is essential for better separation of solid and liquid phase of modified samples. Separation of calcium as a precipitated non-soluble oxalate enables the elimination of spectral interferences originating from different calcium containing species like (40)Ca(35)Cl(+), (40)Ca(37)Cl(+), (43)Ca(16)O(+), (40)Ca(18)O(+), (44)Ca(16)O(+), (43)Ca(16)O(1)H(+) onto the determination of As, Se, Co and Ni whose assay is more difficult when using conventional quadrupole instruments. High detection capability is further an advantage as the approach enables the analysis without dilution. The methodology may serve, in addition, for a fast and sensitive determination of some other elements. After that, direct, reliable and simultaneous determination of 16 elements (Li, Be, B, V, Cr, Mn, Ni, Co, Ga, As, Se, Mo, Sn, Sb, Cs, Tl) at trace and ultra-trace levels in milk can be performed under optimum instrumental conditions and by using Rh as an internal standard. Accuracy and precision was assessed by measuring NCS ZC73015 milk powder control standard, yielding results in agreement with certified values and RSD <10%. The accuracy was also checked by comparison of the results of the proposed method with those found by a method based on a microwave-assisted digestion of real samples.

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