Latest estimates indicate that world prevalence of anti-HCV reactivity
spans widely from 0.1% in developed country to 13,6% in north Africa,
while in Europe prevalence ranges from 0.1% to 4.9% in ...some eastern
European countries. Epidemiological studies show, around 170 million
people in world to be chronically infected with hepatitis C virus
(HCV). Preliminary testing of general population and according to the
estimates, Croatia has classified within countries with HCV prevalence
of 1%-2%, and incidence rate of 0.005%. Chronic HCV infection
represents increasingly significant public health problem, especially
when we take in consideration blood donors as a possible source of
infection. Modern transfusion medicine has an assignment to treat
patients with safe and quality blood preparations while minimizing risk
of infection with blood transmitted diseases. In Croatia all blood
donations are mandatory tested for HIV, HCV, HBV and syphilis. Although
posttransfusion hepatitis C (PTHC) is only one of the possible
infections transmitted by blood, there is no reason to be more concern
than in other countries of Western Europe, where risk of HCV
transmission through blood and blood preparations averages from
1:400.000 to 1:800.000. Despite blood testing PTHC is still possible
complication of transfusion treatments in Croatia and worldwide.
Reasons for that are absence of symptoms and clinical signs of
hepatitis C in incubation period of infected blood donors, absence of
ideal test who would detect all cases of infected blood
The World Health Organization recognized hepatitis C (HCV) as a global
health problem estimated that in 1999, over 170 million people were
chronically infected with HCV. HCV is leading cause of ...end-stage liver
disease and hepatocellular carcinoma. The discovery of hepatitis C
virus (HCV) in 1989 using molecular biology methods has led to the
rapid evolution of the field of HCV diagnostics. Diagnostic tests for
HCV can be divided into serological assays that detect antibody to HCV
and molecular assays that detect, quantify and characterize HCV-RNA
genome within infected patient. Qualitative molecular nucleic acid
tests are used for confirmation of HCV infection and for screening
blood donation. Quantitative HCV-RNA tests provide prognostic
information for monitoring the response to antiviral therapy. HCV is
heterogeneous virus with six distinct genotypes and numerous subtypes.
HCV genotype tests are important clinically because they predict most
accurately the chance of antiviral response and are routinely used for
selecting treatment regimens regarding the duration of interferon
therapy and ribavirin dosage.
SAŽETAK
Svjetska zdravstvena organizacija (WHO) prepoznala je hepatitis C kao svjetski problem. Na osnovi procjena iz godine 1999., 170 milijuna ljudi bilo je kronično inficirano hepatitis C virusom ...(HCV). HCV vodeći je čimbenik neizlječivih oboljenja jetre, uključujući karcinom jetre. Za otkriće HCV-a godine 1989., zaslužni su molekularno-biološki postupci na kojima se zasniva suvremena dijagnostika hepatitisa C. Dijagnostički testovi dijele se na serološke testove kojima se dokazuju
anti-HCV protutijela, te na molekularne testove kojima se u inficirane osobe dokazuje, određuje i obilježuje HCV-RNA genom. Kvalitativni molekularni testovi primjenjuju se za potvrđivanje infekcije, te u kontroli darivane krvi. Praćenjem broja virusnih kopija, kvantitativni HCV-RNA testovi daju podatke o odgovoru na antivirusnu terapiju. HCV jest heterogeni virus koji se na osnovi genomske promjenljivosti svrstava u 6 osnovnih genotipova, te u više podtipova. HCV genotipizacija bitna je za kliničara, budući da se prema pojedinome genotipu može procijeniti odgovor na antivirusnu terapiju, a određivanjem HCV genotipa odabire se optimalni terapijski postupak vezan uz duljinu razdoblja terapije i dozu ribavirina.
SAŽETAK
Najnovije procjene upućuju na to da se svjetska prevalencija anti-HCV reaktivnosti kreće u široku rasponu, i to od 0,1% u razvijenim zemljama do 13,6% u sjevernoj Africi. U Europi ...prevalencija se kreće od 0,1% do najviše 4,9% u pojedinim zemljama istočne Europe. Epidemiološka istraživanja predviđaju da je približno 170 milijuna ljudi u svijetu kronično inficirano hepatitis C virusom (HCV). Na osnovi preliminarnih testiranja opće populacije i prema procjenama, Hrvatska je svrstana u zemlje s nižom HCV prevalencijom (1% – 2%), te incidencijom od 0,005%. Kronična infekcija
HCV-om sve je važniji javnozdravstveni problem, poglavito kada se radi o dobrovoljnim davateljima krvi kao mogućim izvorima zaraze. Zadaća je suvremene transfuziologije liječiti bolesnika sigurnim i kvalitetnim krvnim pripravcima, a postojeći ostatni rizik za infekciju krvlju prenosivim bolestima, svesti na najmanju mjeru. U Hrvatskoj sva se darivanja krvi obvezno testiraju na HIV, HCV, HBV i sifilis. Poslijetransfuzijski hepatitis C (PTHC) jest krvlju prenosiva bolest, te je unatoč obveznu anti-HCV testiranju krvi, još uvijek moguća komplikacija transfuzijskoga liječenja u nas i u svijetu. U zemljama zapadne Europe rizik za prijenos HCV-a
krvlju i krvnim pripravcima kreće se u prosjeku od 1:400.000 do 1:800.000. Razloga je tomu više: od nepostojanja simptoma i kliničkih znakova hepatitisa C u davatelja koji se nalazi u fazi inkubacije, nepostojanja idealnoga testa kojim bi se otkrili mnogobrojni podtipovi HCV virusa, te – iako u malomu postotku, postojanje seronegativnih trajnih nositelja virusa koji su doživotni kliconoše.
Diversity of mitochondrial DNA (mtDNA) lineages of the Island of Cres was determined by high-resolution phylogenetic analysis on a sample of 119 adult unrelated individuals from eight settlements. ...The composition of mtDNA pool of this Island population is in contrast with other Croatian and European populations. The analysis revealed the highest frequency of haplogroup U (29.4%) with the predominance of one single lineage of subhaplogroup U2e (20.2%). Haplogroup H is the second most prevalent one with only 27.7%. Other very interesting features of contemporary Island population are extremely low frequency of haplogroup J (only 0.84%), and much higher frequency of haplogroup W (12.6%) comparing to other Croatian and European populations. Especially interesting finding is a strikingly higher frequency of haplogroup N1a (9.24%) presented with African/south Asian branch almost absent in Europeans, while its European sister-branch, proved to be highly prevalent among Neolithic farmers, is present in contemporary Europeans with only 0.2%. Haplotype analysis revealed that only five mtDNA lineages account for almost 50% of maternal genetic heritage of this island and they present supposed founder lineages. All presented findings confirm that genetic drift, especially founder effect, has played significant role in shaping genetic composition of the isolated population of the Island of Cres. Due to presented data contemporary population of Cres Island can be considered as genetic "outlier" among Croatian populations.
The role of angiogenesis in the pathogenesis of renal cell carcinoma is well recognized, however, the influence of tumor cells in this activity has not yet been fully clarified. The aim of this study ...was to analyze the expression of hypoxia inducible factor-1alpha (HIF-1alpha), a regulatory factor of angiogenic switch, in comparison to vascular endothelial growth factor A and C (VEGF-A and VEGF-C), recognized to be involved in blood and lymph vessel neoangiogenesis, with potential association in the prognosis of patients with renal cell carcinoma.
Ninety-four patients with diagnosis of clear cell renal cell carcinomas (CCRCC), all clinicopathological characteristics and overall survival were unrolled in this study. Immunohistochemicaly VEGF-A, VEGF-C, HIF-1alpha and Ki67 were detected on tumor cells and the staining was performed on tissue microarrays (TMA). The staining was evaluated as a percentage of cytoplasmic or nuclear positive tumor cells.
Variable expression of all three proteins was confirmed. Both angiogenic factors demonstrated perimembranous or diffuse cytoplasmic staining, with diffuse pattern positively associated (p < 0.001). Nuclear HIF-1alpha expression (nHIF-1alpha) showed inverse correlation with diffuse cytoplasmic VEGF-A (p = 0.002) and VEGF-C (p = 0.053), while cytoplasmic HIF-1alpha expression (cHIF-1alpha) showed positive correlation with diffuse staining of both angiogenic factors (p < 0.001; p < 0.001, respectively). In comparison to clinicopathological characteristics, a higher nuclear grade (p = 0.006; p < 0.001, respectively), larger tumor size (p = 0.009; p = 0.015, respectively), higher stage (p = 0.023; p = 0.027, respectively) and shorter survival (p = 0.018; p = 0.024, respectively) were associated with overexpression of cHIF-1alpha and diffuse cytoplasmic VEGF-A expression. In contrary, overexpression of nHIF-1alpha was associated with better diagnostic parameters i.e. lower nuclear grade (p = 0.006), smaller tumor size (p = 0.057), and longer survival (p = 0.005).
Overexpression of VEGF-A and cHIF-1alpha in tumor cells highlights a more aggressive subtype of CCRCC that might have some clinical implications. The significance of nHIF-1alpha expression associated with better differentiated tumors should be further elucidated.
The aim of this study is to investigate the differences of clinical and laboratory parameters between patients with JAK2-V617F positive myeloproliferative neoplasms (MPNs) and JAK2 wild type MPNs. ...DNA was isolated from peripheral blood granulocytes of 106 patients treated at Rijeka University Hospital Center: 41 with polycythemia vera (PV), 43 with essential thrombocythemia (ET), 9 with primary myelofibrosis (PMF) and 13 with myeloproliferative neoplasm--unclassifiable (MPN-u). The JAK2-V617F mutation was detected using allele specific PCR. Laboratory and clinical parameters were obtained from patient's medical records. The JAK2-V617F mutation was detected in 69% (73/106) patients with MPNs. The results revealed significantly different prevalence of JAK2-V617F mutation, between MPNs entities: 88% in PV 58% in ET, 56% in PMF and 54% in MPNs-unclassified disorders. The JAK2-V617F mutation significantly correlated with higher leukocyte count and alkaline phosphatase co re in ET group and with higher platelets count, leukocyte alkaline phosphatase score and serum lactate dehydrogenase in PV group. Vascular events were associated with elevated platelets count in whole MPNs group, with higher platelets and leukocyte count in ET and with splenomegaly in PVpatients. Clinical and laboratory data revealed significant contribution ofJAK2-V617F mutation to the development of clinical phenotype in patients with distinct subgroups of MPNs.
The study searched for epidermal growth factor receptor (EGFR) gene amplification in hyperplastic glottis lesions. After classical pathohistological findings of hematoxylin-eosin (HE) slides and ...quantitative immunohistochemical (IHC) analysis, fluorescent in situ hybridization (FISH) was used on tissue microarrays of laryngeal hyperplastic tissue ranging from normal mucosa to abnormal and atypical hyperplastic lesions. FISH analysis of two atypical hyperplastic lesions discovered the amplification of EGFR gene while it was not found in simple and abnormal hyperplastic lesions. The results may indicate that EGFR gene amplifications could possibly correlate with the histopathologic picture. Tissue samples burdened with specific oncogen signatures like EGFR gene amplification could be detected in precancerous lesion. This might improve follow-up and treatment protocols of glottic lesions which are an everyday problem for ENT practitioners. Further research is mandatory to confirm our findings.
The DRB region of the human major histocompatibility complex displays length polymorphism: Five major haplotypes differing in the number and type of genes they contain have been identified, each at ...appreciable frequency. In an attempt to determine whether this haplotype polymorphism, like the allelic polymorphism, predates the divergence of humans from great apes, we have worked out the organization of the DRB region of the chimpanzee Hugo using a combination of chromosome walking, pulsed-field gel electrophoresis, and sequencing. Hugo is a DRB homozygote whose single DRB haplotype is some 440 kilobases (kb) long and contains five genes. At least one and possibly two of these are pseudogenes, while three are presumably active genes. The genes are designated DRB*A0201, DRB2*0101, DRB3*0201, DRB6*0105, and DRB5*0301, and are arranged in this order on the chromosome. The DRB2 and DRB3 genes are separated by approximately 250 kb of sequence that does not seem to contain any additional DRB genes. The DRB*A0201 gene is related to the DRB1 gene of the human DR2 haplotype; the DRB2*0101 and DRB3*0201 genes are related to the DRB2 and DRB3 genes of the human DR3 haplotype, respectively; the DRB6*0105 and DRB5*0301 genes are related to the DRBVI and DRB5 genes of the human DR2 haplotype, respectively. Thus the Hugo haplotype appears to correspond to the entire human DR2 haplotype, into which a region representing a portion of the human DR3 haplotype has been inserted. Since other chimpanzees have their DRB regions organized in different ways, we conclude that, first, the chimpanzee DRB region, like the human DRB region, displays length polymorphism; second, some chimpanzee DRB haplotypes are longer than the longest known human DRB haplotypes; third, in some chimpanzee haplotypes at least, the DRB genes occur in combinations different from those of the human haplotypes; fourth, and most importantly, certain DRB gene combinations have been conserved in the evolution of chimpanzees and humans from their common ancestors. These data thus provide evidence that not only allelic but also haplotype polymorphism can be passed on from one species to another in a given evolutionary lineage.
To isolate and genotype Borrelia burgdorferi genospecies in serum samples of Croatian patients with erythema migrans.
DNA isolates from sera of patients with erythema migrans were analyzed by nested ...polymerase chain reaction (PCR), amplifying a segment of flagellin gene with primers encompassing the conserved region of the gene. To screen PCR products for heterogeneity, we performed single-stranded conformation polymorphism (SSCP) analysis. The samples showing differences in SSCP patterns were sequenced, and the sequence compared in the GeneBank for sequence homology with known Borrelia burgdorferi genospecies. We also constructed phylogenetic tree of all known borrelial sequences.
The nested PCR method using specially designed flagellin gene primers, achieved the sensitivity of 10 genome copies (0.01 pg of purified Borrelia burgdorferi DNA from culture) by dilution analysis. The assay specificity was confirmed by amplification of a part of the flagellin gene from different bacterial species. The primer pairs successfully amplified only Borrelia burgdorferi flagellin gene. The genome of Borrelia burgdorferi sensu lato was detected in the sera of all 10 tested patients with erythema migrans. Sequence data and phylogenetic analysis confirmed that all amplified samples belonged to Borrelia afzelii genospecies.
Phylogenetic tree analysis placed the borrelial isolates together with Borrelia afzelii sequences into a single group. This finding was additionally supported by sequence homology analysis, which produced a homology score of 99%. In patients with erythema migrans who come from the northwest Croatia, an endemic area for Lyme borreliosis, Borrelia afzelii was the cause of skin manifestations of Lyme borreliosis.
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