[Clinical progression of the first wave of novel coronavirus infection in Ostrava].

OBJECTIVES: The first case of coronavirus infection in the Moravian-Silesian Region was diagnosed on March 12, 2020. The study aimed to describe the first wave and clinical manifestation of the coronavirus epidemic at the Department of Infectious Diseases in Ostrava. MATERIAL AND METHODS: The sample comprised a total of 195 patients requiring hospitalization at the Department of Infectious Diseases in Ostrava between March 1 and August 31, 2020. The virus was diagnosed using polymerase chain reaction from nasopharyngeal swabs in 192 patients and from the bronchoalveolar lavage in one patient. In the other two patients, serological tests were applied using virus neutralization assays and ELISA specific antibodies. RESULTS: The sample included 100 men and 95 women. The mean age was 69.5 years. The most common diagnosis was pneumonia, observed in 123 patients (63 %). Respiratory symptoms without pneumonia were seen in 37 patients (19 %); loss of smell and taste in 14 patients (7 %); cardiovascular complications in 9 patients (5 %); acute psychosis in 2 patients (1 %); and diarrhea and vomiting in 25 patients (13 %). The remaining 23 patients (12 %) did not present any symptoms. The mean length of hospital stay was 11 days. The most frequent comorbidity was cardiovascular disease (70 %). A total of 39 patients died (20 %); their mean age was 77 years. Mechanical ventilation was started in 16 patients, of whom 7 died (43 %). The treatment was mostly symptomatic. Hydroxychloroquine was administered to 21 patients (11 %), favipiravir to 26 patients (13 %) and remdesivir to 13 patients (6 %). CONCLUSION: Respiratory complications predominated in the majority of patients with coronavirus infection. In most cases, the treatment was symptomatic. One-fifth of the patients, mostly elderly ones, died.

[Two case reports of chronic hepatitis C retreatment].

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.

[Use of PCR for diagnosing cat-scratch disease].

OBJECTIVE: The diagnosis of Bartonella henselae by polymerase chain reaction (PCR) in lymph nodes removed in 10 patients with serologically confirmed evidence cat-scratch disease. MATERIAL AND METHODS: The 2015-2018 group consisted of 10 patients with serologically confirmed cat-scratch disease, all of them having positive IgG antibodies and 6 patients also positive IgM antibodies against B. henselae. The group included 4 men and 6 women, 7 children and 3 adults, aged 5-52 years. Eleven lymph nodes obtained from the 10 patients were formalin-fixed paraffin-embedded. Variants of granulomatous inflammation were found in 9 patients; a 13-year-old boy had Hodgkin's lymphoma. DNA isolation was performed with cobas® DNA Sample Preparation Kit (Roche). DNA of Bartonella spp. was detected by real-time PCR with BactoReal® Kit Bartonella spp. (Ingenetix) detecting the gltA gene specific for the genus Bartonella. RESULTS: Four of the 10 patients tested positive or borderline positive for Bartonella when their histological material was analyzed by PCR. One patient with 2 lymph nodes examined showed a positive result for only 1 lymph node. One adult male had a positive result; three children showed borderline positive results. Of those, two patients had suppurative granulomatous and the other 2 patients had necrotizing suppurative granulomatous inflammation as histological findings. All 4 patients had positive IgM antibodies against B. henselae. The boy with lymphoma had a negative PCR result. CONCLUSION: Serological tests combined with histological examination of lymph nodes and PCR may improve the diagnosis of cat- scratch disease.

Lymph node syndrome associated with cat scratch disease in children and adults.

A retrospective evaluation of a group of patients with cat-scratch disease which is focused on epidemiological context, clinical picture, serological diagnosis and treatment in children and adults. Seventy-three patients with catch-scratch disease (CSD) were diagnosed at the Clinic of infectious medicine in 2008-2017, the group consisted of 37 children and 36 adults. Injury, or at least contact with cat, localization of lymph node syndrome and other clinical signs were investigated. Serological diagnosis was based on detection of IgM and IgG antibodies against Bartonella henselae. Duration of lymph node syndrome and antibodies against B. henselae were observed. Findings of granulomatous inflammation in extirpated lymph node supported suspicion of CSD. 66 of 73 patients had contact with cat, only 22 patients reported injury caused by cat. Cervical lymph nodes were affected the most often, in 32 patients. Extirpation of lymph node was performed in 19 patients. Positive IgM antibodies were detected in 47 patients. Between the group of children and adults important differences in any followed parameters were not observed. In the group of 73 patients with cat-scratch disease cervical lymph nodes were affected the most often. Positive IgM antibodies were detected in 47 patients, lymph node extirpation was performed in 19 patients. Considerable differences in course of disease in children and adults were not found out. Keywords: cat-scratch disease, lymph node syndrome, serology, extirpation.

Characterization of Hepatitis C Virus IRES Quasispecies - From the Individual to the Pool.

Hepatitis C virus (HCV) is a single-stranded positive-sense RNA virus from the genus Hepacivirus. The viral genomic +RNA is 9.6 kb long and contains highly structured 5' and 3' untranslated regions (UTRs) and codes for a single large polyprotein, which is co- and post-translationally processed by viral and cellular proteases into at least 11 different polypeptides. Most of the 5' UTR and an initial part of the polyprotein gene are occupied by an internal ribosome entry site (IRES), which mediates cap-independent translation of the viral proteins and allows the virus to overcome cellular antiviral defense based on the overall reduction of the cap-dependent translation initiation. We reconsidered published results concerning a search for possible correlation between patient response to interferon-based antiviral therapy and accumulation of nucleotide changes within the HCV IRES. However, we were unable to identify any such correlation. Rather than searching for individual mutations, we suggest to focus on determination of individual and collective activities of the HCV IRESs found in patient specimens. We developed a combined, fast, and undemanding approach based on high-throughput cloning of the HCV IRES species to a bicistronic plasmid followed by determination of the HCV IRES activity by flow cytometry. This approach can be adjusted for measurement of the individual HCV IRES activity and for estimation of the aggregate ability of the whole HCV population present in the specimen to synthesize viral proteins. To detect nucleotide variations in the individual IRESs, we used denaturing gradient gel electrophoresis (DGGE) analysis that greatly improved identification and classification of HCV IRES variants in the sample. We suggest that determination of the collective activity of the majority of HCV IRES variants present in one patient specimen in a given time represents possible functional relations among variant sequences within the complex population of viral quasispecies better than bare information about their nucleotide sequences. A similar approach might be used for monitoring of sequence variations in quasispecies populations of other RNA viruses in all cases when changes in primary sequence represent changes in measurable and easily quantifiable phenotypes.

[Vertical hepatitis C transmission - authors' experiences].

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.

[Guideline for the diagnosis and treatment of acute bacterial meningitis].

Acute bacterial meningitis is a severe infectious disease of the central nervous system. Its incidence decreases but lethality and sequelae remain high. The early initiation of appropriate treatment is a factor strongly determining the patient´s prognosis. The authors submit the Czech national guideline for diagnosis and treatment of community-acquired acute bacterial meningitis which has to provide clear and simple recommendations for clinicans involved in the care of meningitis in adults and children. The national guideline was based on the European guideline published in 2016 and adapted for the situation in the Czech Republic. It was acknowledged (approved? ratified?) by the Society for Epidemiology and Microbiology and the Society for Medical Microbiology of the Czech Medical Association.

[Diagnosis and therapy of hepatitis B virus infection: Czech national guidelines].

The new recommendations reflect the increase in knowledge that has been reported since the release of previous Czech guidelines in September 2014. The basis for these guidelines were the European Association for the Study of the Liver guidelines from April 2017. According to qualified estimates, there are 240 million people with chronic hepatitis B (HBV) infection worldwide. The Czech Republic is among the countries with a low prevalence of HBV infection. According to the latest seroprevalence study, 0.56 % of the Czech citizens were chronically infected with HBV in 2001. A similar study conducted in only two regions of the Czech Republic in 2013 showed a prevalence of only 0.064 %. HBV infection can lead to serious life-threatening liver damage - fulminant hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC). The main goals of treatment are to prolong the length of life and improve its quality by preventing the progression of chronic hepatitis to cirrhosis, cirrhosis decompensation and development of HCC. The goals may be achieved if HBV replication is suppressed in a sustained manner. Additional goals are prevention of vertical transmission from mother to newborn, inhibition of HBV reactivation and therapy of HBV-related extrahepatic manifestations. Generally, there are two different strategies of chronic hepatitis B therapy available - treatment with nucleoside or nucleotide inhibitors (NIs) or with pegylated interferon alfa. Currently, the vast majority of Czech and European patients are treated with NIs. The NIs that have been approved for HBV treatment in the European Union include lamivudine, adefovir dipivoxil, entecavir (ETV), telbivudin (TBV), tenofovir disoproxil fumarate (TDF) and tenofovir alafenamide (TAF). TAF and TBV have not yet been marketed in the Czech Republic. The main advantages of treatment with potent NIs with a high barrier to resistance (ETV, TDF, TAF) are their predictable high long-term antiviral efficacy leading to undetectable HBV DNA levels in the vast majority of compliant patients as well as their favorable safety profiles. These drugs can be used in any HBV infected patient and represent the only treatment option for patients with decompensated liver cirrhosis, liver transplants, extrahepatic HBV-related manifestations, severe acute hepatitis B or chronic HBV reactivation.

[Investigation of autoimmunity markers during interferon alpha therapy of chronic hepatitis B and C - twenty years of experience]. / Sledování autoimunitnich fenoménu pri lécbe chronické hepatitidy B a C interferonem alfa - dvacetileté zkusenosti.

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.

[Furuncular myiasis - 2 case reports]. / Furunkulózní myiaza- 2 kazuistiky.

Furuncular myiasis caused by the larvae of the human botfly, Dermatobia hominis, is a common disease in tropical areas of Latin America and may therefore occur in travelers returning from that region. Outside the endemic areas, the diagnosis is often delayed because the disease is mistaken for a bacterial skin infection. Reported are two cases of furuncular myiasis in patients returning from Central and South America.

[Thyroid dysfunction during interferon alpha therapy for chronic hepatitis B and C - twenty years of experience].

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.

Anti-HBs antibody persistence following primary vaccination with an investigational AS02(v)-adjuvanted hepatitis B vaccine in patients with renal insufficiency.

BACKGROUND: Three doses of the investigational AS02(v)-adjuvanted hepatitis B virus (HBV) vaccine HB-AS02 have been shown to induce more rapid seroprotection and higher anti-HBs antibody concentrations in patients with renal insufficiency than four doses of FENDrix™ (HB-AS04), an adjuvanted HBV vaccine licensed in Europe for use in this population. This study evaluated persistence of immune response up to 36 months after primary vaccination. METHODS: In this open, international, Phase III follow-up study, 151 pre-dialysis, peritoneal dialysis and hemodialysis patients ≥15 years of age received HB-AS02 at 0, 1, 6 months and 149 received HB-AS04 at 0, 1, 2, 6 months. Of these, 99 and 80 returned at Month 36, 76 and 62 of whom were eligible for inclusion in the Long-Term According-To-Protocol (LT-ATP) cohort for descriptive analysis of antibody persistence (mean age: 65.6 years). RESULTS: At Month 36, 89.5% of subjects in the HB-AS02 group and 72.6% of those in the HB-AS04 group had anti-HBs antibody concentrations ≥10 mIU/ml. Anti-HBs antibody concentrations were ≥100 mIU/ml in 82.9% and 35.5% of subjects, respectively. Anti-HBs geometric mean antibody concentrations were higher in the HB-AS02 group over the 36 months of follow-up. An exploratory "time to boost" analysis confirmed that subjects who received HB-AS02 were 2.54 times more likely than those who received HB-AS04 to have anti-HBs antibody concentrations ≥10 mIU/ml at Month 36 (p=0.013 [95% CI: 1.22, 5.31]). CONCLUSION: HB-AS02 candidate vaccine induces high and persistent anti-HBs antibody levels in pre-dialysis, peritoneal dialysis and hemodialysis patients, potentially reducing the need for booster doses in this population.

[Bronchobiliary fistulae as a complication of hepatic cystic echinococcosis]. / Bronchobiliární pístel jako komplikace echinokokové cysty jater.

Cystic hydatid disease or cystic echinococcosis (CE) rarely occurs in the Czech Republic. In 2005 - 2009, eleven cases were recorded, mostly among immigrants from the Balkans. Presented here is a case report of a 38-year-old patient with hepatic CE complicated by bronchobiliary fistulae. Ten days before surgical removal of the hydatid cysts, treatment with mebendazole was started. During surgery the affected part of the lungs was resected and the liver cysts were drained using transthoratic access. The follow-up was complicated by leakage of bile into the pleural cavity. The leakage was associated with continued communication between the liver cyst and the pleural cavity which did not close spontaneously after removal of the drain. Endoscopic nasobiliary drainage decreased pressure in the bile duct and within 14 days, it led to the spontaneous closure of the communication between the liver cyst and the pleural cavity. Seven months after the operation, the patient was in a very good clinical condition.

[Low response rate to a vaccination against hepatitis B in patients with end-stage liver disease]. / Malá úcinnost vakcinace proti virové hepatitide B u pacientu s pokrocilým jaterním onemocnením.

Hepatitis B vaccination was started in 41 patients with end-stage liver disease who were liver transplant candidates. Patients received three 20 microg or, starting from 1999, 40 microg doses of recombinant vaccine at 0, 2 and 4 weeks. Blood samples were obtained 4 weeks after vaccination or each revaccination; patients without protective hepatitis B surface antibodies (anti-HBs) were once or repeatedly revaccinated. Thirty-eight patients received at least 3 doses of vaccine. Protective anti-HBs level (≥ 10 IU/l) was detected in 17 of 34 patients (50 %) after the third up to eight dose. No case of chronic HBsAg carrier status was detected. Immunisation against hepatitis B in persons with liver cirrhosis is associated with a poor response and new vaccines should be considered for these patients.

[Haemophilus influenzae meningitis in a child with complete immunisation]. / Hemofilová meningitida u rádne ockovaného dítete.

Invasive Haemophilus influenzae b (Hib) diseases are rarely observed in children with vaccination against Hib. Immunisation of infants against Hib was introduced in the Czech Republic in 2001. The first case of meningitis with isolation of Hib in child immunised with Hib vaccine since starting routine vaccination is described. Clinical course of meningitis was mild and was identical with previously manifestation of Hib meningitis.