Diagnosis and treatment of Bartonella endocarditis.

The Bartonella genus comprises more than 20 species of Gram-negative rods which are difficult to culture. These are facultative intracellular bacteria. Humans are reservoir hosts for B. quintana and B. bacilliformis or accidental hosts for other species. Bartonella is a cause of zoonosis. Bartonella infection can be completely asymptomatic or can be linked to various conditions. Our experience with Bartonella endocarditis from 2012-2017 is presented. The most effective diagnostic method for Bartonella endocarditis is PCR detection of DNA of the pathogen from excised valve tissue. The European Society of Cardiology (ESC) in the guidelines from 2015 recommends the combination doxycycline gentamycin for the treatment of Bartonella endocarditis.

[The use of IgG antibody avidity assays in the diagnosis of cytomegalovirus infection]. / Vyuzití testu avidity IgG protilátek v diagnostice cytomegalovirové infekce.

Human cytomegalovirus (CMV) is the most common cause of congenital infection. Primary CMV infection can lead to severe disease and complications in patients immunocompromised as a result of disease or therapy. IgG antibody avidity assays make it possible to differentiate between primary infection and reactivation of latent infection or reinfection. The study objective was to determine CMV IgG avidity by enzyme-linked immunosorbent assay (ELISA) with denaturation of IgG antibody binding to the antigen and by chemiluminiscent microparticle immunoassay (CMIA) on an Abbott Architect analyzer. Both methods yielded comparable CMV IgG avidity results. In some cases, the Abbott test was superior in reflecting IgG antibody maturation during primary infection to microplate ELISA using antigen-antibody complex dissociation by a denaturing agent.

The effect of rat strain, diet composition and feeding period on the development of a nutritional model of non-alcoholic fatty liver disease in rats.

Non-alcoholic fatty liver disease (NAFLD) is an important cause of liver-related morbidity and mortality. The aim of this work was to establish and characterize a nutritional model of NAFLD in rats. Wistar or Sprague-Dawley male rats were fed ad libitum a standard diet (ST-1, 10 % kcal fat), a medium-fat gelled diet (MFGD, 35 % kcal fat) and a high-fat gelled diet (HFGD, 71 % kcal fat) for 3 or 6 weeks. We examined the serum biochemistry, the hepatic malondialdehyde, reduced glutathione (GSH) and cytokine concentration, the respiration of liver mitochondria, the expression of uncoupling protein-2 (UCP-2) mRNA in the liver and histopathological samples. Feeding with MFGD and HFGD in Wistar rats or HFGD in Sprague-Dawley rats induced small-droplet or mixed steatosis without focal inflammation or necrosis. Compared to the standard diet, there were no significant differences in serum biochemical parameters, except lower concentrations of triacylglycerols in HFGD and MFGD groups. Liver GSH was decreased in rats fed HFGD for 3 weeks in comparison with ST-1. Higher hepatic malondialdehyde was found in both strains of rats fed HFGD for 6 weeks and in Sprague-Dawley groups using MFGD or HFGD for 3 weeks vs. the standard diet. Expression of UCP-2 mRNA was increased in Wistar rats fed MFGD and HFGD for 6 weeks and in Sprague-Dawley rats using HFGD for 6 weeks compared to ST-1. The present study showed that male Wistar and Sprague-Dawley rats fed by HFGD developed comparable simple steatosis without signs of progression to non-alcoholic steatohepatitis under our experimental conditions.

[Difficulties in laboratory diagnosis of cerebral toxoplasmosis]. / Uskalí laboratorní diagnostiky mozkové toxoplazmózy.

Toxoplasmosis is the most wide-spread parasitic disease in the Czech Republic. According to the results of serological studies, about 25-50% of its population come in contact with this protozoan. A serious form of the disease may develop in severely immunocompromised patients. In these patients, problems with diagnosing toxoplasmosis may occur, especially in the case of its rare but serious cerebral form. The aim of the case report is to present potential difficulties in the diagnosis of cerebral toxoplasmosis.

[Amniotic fluid interleukin 6 levels in preterm premature rupture of membranes]. / Interleukin 6 v plodové vode pacientek s predcasným odtokem plodové vody.

OBJECTIVE: The purpose of this study was to determinate the changes of amniotic fluid interleukin 6 (IL-6) concentrations in patients with preterm premature rupture of the membranes (PPROM), and in the presence of microbial invasion of the amniotic cavity (MIAC) and histological chorioamnionitis (HCA). The aim was to examine amniotic fluid IL-6 in relation to MIAC and HCA. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynecology Medical Faculty Charles University Hradec Králové. METHODS: We studied 37 women between 24 and 36 weeks of gestation with PPROM. Samples of amniotic fluid were collected by transabdominal amniocentesis. Polymerase chain reaction for the genital mycoplasmas and culture for aerobic and anaerobic bacteria were performed. Twenty-eight of 37 patients placentas were collected and assessed for presence or absence HCA. IL-6 concentration in amniotic fluid were determined using a sensitive and specific diagnostic kit Human IL-6 Quantikine ELISA manufactured R&D Systems, USA. RESULTS: There was significant difference in the median amniotic fluid IL-6 concentration between patients with preterm rupture of the membranes with and without MIAC and HCA (patients with MIAC and HCA: median 915 pg/ml, range 651-1854 pg/ml vs. patients without MIAC and HCA: median 780 pg/ml, range 184-1059 pg/ml; p=0.047). There was no significant difference in the median amniotic fluid IL-6 concentration between patients with preterm rupture of the membranes with and without MIAC (patients with MIAC: median 915 pg/ml, range 195-1854 pg/ml vs. patients without MIAC: median 792 pg/ml, range 184-1993 pg/ml; p=0.53). There was no significant difference in the median amniotic fluid IL-6 concentration between patients with preterm rupture of the membranes with and without HCA (patients with HCA: median 829 pg/ml, range 195-1992 pg/ml vs. patients without HCA: median 768 pg/ml, range 184-1890 pg/ml; p = 0.31). CONCLUSION: Amniotic fluid IL-6 concentrations patients with PPROM with presence HCA and MIAC were significantly higher than IL-6 concentration patients without HCA and MIAC.

Primer R108 performs best in the RAPD strain typing of three Aspergillus species frequently isolated from patients.

We evaluated the suitability of various primers for the RAPD (random amplified polymorphic DNA) accurate species identification and strain typing of Aspergillus clinical isolates. Five primers described previously were tested for their discriminatory power in three Aspergillus species (A. fumigatus, A. niger agg. and A. flavus - 23 clinical isolates and 2 reference strains). Clustering of RAPD fingerprints corresponded well with the identification based on morphological features. All isolates were resolved as different strains using the primer R108 and the RAPD protocol optimized for a Robocycler thermal cycler. RAPD with the primer R108 thus can be considered to be a valuable, simple and powerful tool for identification and strain delineation of Aspergillus spp.

[Mumps--a reemerging infection? The current incidence of mumps in the East Bohemian region in the Czech Republic]. / Príusnice--znovu se vracející infekce? Soucasná situace ve výskytu príusnic ve východoceském regionu.

A review of mumps outbreaks among both non-vaccinated and vaccinated children and young adults in the East Bohemian region in 2003-2005 is presented. A significant increase in mumps cases was observed over this period. The clinical diagnosis was confirmed serologically by ELISA detection of IgM antibodies and/or IgG seroconversion and increased levels of IgG antibodies. A reverse transcriptase nested PCR was introduced for direct detection of mumps virus RNA from clinical specimens (nasopharyngeal secretion, saliva, CSF and serum). The isolated RNA will be stored for further analysis and mumps virus genotyping attempts, helpful in tracing the virus circulation in the East Bohemia region. Possible causes of the recent significant increase in mumps cases among the vaccinated population in the Czech Republic are discussed.

Time-course of hormonal induction of mitochondrial glycerophosphate dehydrogenase biogenesis in rat liver.

Thyroid hormones are important regulators of mitochondrial metabolism. Due to their complex mechanism of action, the timescale of different responses varies from minutes to days. In this work, we studied selective T3 induction of the inner mitochondrial membrane enzyme-glycerophosphate dehydrogenase (mGPDH) in liver of euthyroid rats. We correlated the kinetics of the T3 level in blood, the mRNA level in liver, the activity and amount of mGPDH in liver mitochondria after a single dose of T3. The T3 level reached maximum after 1 h (80 nmol/l) and subsequently rapidly decreased. mGPDH mRNA increased also relatively fast, reaching a maximum after 12 h and fell to the control level after 72 h. An increase of mGPDH activity could be already found after 6 h and reached a maximum after 24 h in accordance with the increase in mGPDH content (2.4-fold vs. 2.7-fold induction). After 72 h, the mGPDH activity showed a significant 30% decrease. When the rats received three subsequent doses of T3, the increase of mGPDH activity was 2-fold higher than after a single T3 dose. The results demonstrate that mGPDH displays rapid induction as well as decay upon disappearance of a hormonal stimulus, indicating a rather short half-life of this inner mitochondrial membrane enzyme.

Glycerophosphate-dependent hydrogen peroxide production by rat liver mitochondria.

We studied the extent to which hormonally-induced mitochondrial glycerophosphate dehydrogenase (mGPDH) activity contributes to the supply of reducing equivalents to the mitochondrial respiratory chain in the rat liver. The activity of glycerophosphate oxidase was compared with those of NADH oxidase and/or succinate oxidase. It was found that triiodothyronine-activated mGPDH represents almost the same capacity for the saturation of the respiratory chain as Complex II. Furthermore, the increase of mGPDH activity induced by triiodothyronine correlated with an increase of capacity for glycerophosphate-dependent hydrogen peroxide production. As a result of hormonal treatment, a 3-fold increase in glycerophosphate-dependent hydrogen peroxide production by liver mitochondria was detected by polarographic and luminometric measurements.