Fatal intoxication by intravenous injection of castor bean (Ricinus communis L.) extract-a case study.

A case report of a 25-year-old man who committed suicide by intravenous injection himself of an aqueous home-made castor bean extract is presented. The patient was hospitalized and treated symptomatically and was released at its own request fourth day after intoxication. The next day, the patient's condition deteriorated, and he died 6 days after intoxication even though he was given medical care. Case history, autopsy, and toxicological investigation of ante- and post-mortem collected materials are described. Blood and urine collected from the patient ante-mortem and other several biological materials (namely blood from the upper and lower limb, blood from the right and left ventricle, pericardial fluid, vitreous humour, liver, kidney, and spleen) were collected post-mortem during autopsy. Liquid-liquid extraction procedure followed by high-performance liquid chromatography tandem mass spectrometry analysis for identification and determination of ricinine as a biomarker of ricin/castor seed intoxication was developed and validated. The method was applied on analysis of collected ante- and post-mortem biological materials. The post-mortem contents of ricinine in organs (namely the liver, kidney, and spleen) are firstly reported. The obtained results indicated approximately uniform distribution of ricinine (concentration level about 1 ng mL-1) in the body after death. In addition, the GC-MS method was also applied for the analysis of extract of castor seed and the patient's urine, to demonstrate alternative possibility for identification of ricinine for clinical and forensic purposes.

Fluid content in the pleural cavity protects internal structures against heat.

PURPOSE: In this paper we present a case report of two people who were found dead in a holiday cottage after a fire. The cause of death in these cases was clarified by the presence of blood in the left pleural cavity in both bodies. METHODS: An inspection of the bodies at the scene of the fire and subsequent examination by a medical examiner suggested the deaths were accidental. RESULTS: An autopsy showed that the bodies were those of a man and a woman with severe carbonization of the skin, soft tissues, brain, and some organs of the thoracic and abdominal cavities. There were also isolated stab injuries to the chest, as well as injuries to skeletal structures and intrathoracic organs. The accumulated blood in the pleural cavity had protected the internal organs against thermal destruction and preserved the traumatic findings. CONCLUSION: To the best of our knowledge this is the first description of the protective effect of hemothorax against thermal damage of the intrathoracic organs.

Prevalence of human leukocyte antigen HLA-B*57:01 in HIV-infected subjects in the Czech Republic.

The HLA-B*57:01 allele is associated with a hypersensitivity reaction to abacavir, and its prevalence varies in different populations. The aim of the study was to investigate HLA-B*57:01 prevalence in the Czech HIV-infected population. HLA-B*57:01 prevalence in our cohort was 5.33%, which is similar to the situation in other Central European countries.

[Our experience with maraviroc treatment in HIV positive patients]. / Nase zkusenosti s maravirocem u HIV pozitivních pacientu.

UNLABELLED: BACKGROUND; Although antiretroviral therapy has changed the clinical course of HIV infection, AIDS remains an incurable disease. Virus entry inhibitors, including maraviroc as the only registered representative of the class, represent a newly emerged group of anti-retrovirals with novel mechanism of action. The primary endpoint is to evaluate the clinical efficacy parameter of maraviroc by measuring viral load at the end of the 4 week treatment period. The secondary endpoint is to evaluate the effectiveness of the drug by monitoring the changes of the viral load values and CD4+ cell counts during the period of 125 weeks. Drug safety was also assessed. METHODS AND RESULTS: Data of 23 subjects were collected, 21 patients were from the Czech Republic and 2 patients from France. Decrease in viral load in the 4th, 24th and 48th week was more than two orders of magnitude (-2.136; -2.448; -2.452 log10 copies/ml). The CD4+ cell count increased (71.71, 143.00, 196.43 cells/mm3). Drug safety was assessed by monitoring the frequency of adverse effects. The data obtained were compared with the III. phase of clinical trials. CONCLUSIONS: Our experience with maraviroc was positive. Maraviroc proved to be an effective antiretroviral agent for a combination therapy of HIV infection.

[Endocrine complications in HIV/AIDS patients]. / Endokrinni komplikace HIV/AIDS infikovaných pacientu.

A number of endocrine abnormalities develop in patients with HIV infection, although many are likely responses to infection, stress, and malnutrition. Others are due to infiltration of endocrine glands by tumor or infection. This article reviews adrenal, testicular, resp.gonadal, pancreatic, calcium and thyroid abnormalities in HIV infected patients.

Isolation and identification of the Amanita muscaria and Amanita pantherina toxins in human urine.

OBJECTIVES: Ibotenic acid, muscimol and muscarine were recognized as responsible for psychotropic effects of A. muscaria and A. pantherina. Demand for their specific and sensitive identification and quantitation in biological material lead to effort to develop reliable analytical method, but satisfactory solution is still lacking. Presented article describes liquid chromatography-mass spectrometry method suitable for isolation and identification of principal toxins of A. muscaria and A. pantherina in urine. METHODS AND RESULTS: Dedicated liquid chromatography-mass spectrometry method is reported. Technique consists of an extraction of analytes on Strata X-CW and Discovery SCX SPE cartridges and separation is achieved using a Gemini C18 column (150 mm x 2.0 mm, 5 micron) and 8 mM heptafluorobutyric acid as mobile phase. Detection at m/z 159 for ibotenic acid, m/z 115 for muscimol and m/z 174 for muscarine was used. Retention times and LODs are 2.6 min and 50 ng.ml-1 for ibotenic acid, 4.6 min and 40 ng.ml-1 for muscimol and 14.2 min and 3 ng.ml-1 for muscarine. CONCLUSION: A sensitive and specific liquid chromatography-mass spectrometry assay was developed for analysis of principal toxins of A. muscaria and A. pantherina in urine. Method was found to be sufficiently sensitive and specific for analysis of urine of intoxicated patients.

A study of HIV-1 genetic diversity in the Czech Republic: 1986-2007.

BACKGROUND: The global HIV/AIDS epidemic consists of a number of regional epidemics caused by different HIV-1 subtypes prevailing in different regions. OBJECTIVES: To study changes in genetic diversity of HIV-1 strains isolated in the Czech Republic (CR) over a more than twenty-year period (1986-2007). STUDY DESIGN: HIV-1 strains isolated in CR from 1986 to 2007 were subtyped by pol gene sequencing followed by phylogenetic analysis. The role of HIV-1 subtyping in molecular epidemiology was considered. RESULTS: Awide range of HIV-1 subtypes were found, with subtype B, into which 76.6% of 534 HIV-1 isolates were classified, being predominant during the whole study period. An increasing number of non-B subtypes A1, C, D, F1, G and some recombinant forms (CRF 01_AE, CRF 02_AG and CRF 06_cpx) were identified after 1990. CONCLUSIONS: The absolute predominance of subtype B among HIV-1 strains in the Czech Republic ended in 1991 when different non-B subtypes had been introduced into the country. The East-West migration is responsible for the introduction of HIV-1 subtypes prevalent in Eastern European and some Asian countries. Genetic analysis of HIV-1 isolates from a given region can be helpful in tracing the course of the HIV/AIDS epidemic.

[Proposals for guidelines for vaccination of adults infected with HIV-1 in the Czech Republic]. / Návrh doporuceného postupu ockování u dospelých osob infikovaných virem HIV-1.

HIV positive patients are in a higher risk of many infections, including the preventable ones. The vaccination is thus a very important part of health-care offered to those patients. Effectiveness of vaccination correlates strongly with the actual immunological status. Vaccination is safe, including some live vaccines, in persons with CD4+ T cells > 500/ml. In patients with CD4+ T cells > 200/ml the efficacy of vaccination is uncertain and live vaccines are strictly contraindicated. A good knowledge of all aspects of vaccination and HIV-infection is necessary, hence we recommend that vaccination of all HIV-positive persons should be realized exclusively by experts in specialized AIDS centers. In the following text we present the proposals for guidelines for vaccination of adult patients infected with HIV-1.

Fatal Ingestion of Chlumsky Disinfectant Solution.

A 32-year-old pregnant woman in the 25th week of pregnancy underwent oral glucose tolerance screening at the diabetologist's. Later that day, she was found dead in her apartment possibly poisoned with Chlumsky disinfectant solution (solutio phenoli camphorata). An autopsy revealed chemical burns in the digestive system. The lungs and the brain showed signs of severe edema. The blood of the woman and fetus was analyzed using gas chromatography with mass spectrometry and revealed phenol, its metabolites (phenyl glucuronide and phenyl sulfate) and camphor. No ethanol was found in the blood samples. Both phenol and camphor are contained in Chlumsky disinfectant solution, which is used for disinfecting surgical equipment in healthcare facilities. Further investigation revealed that the deceased woman had been accidentally administered a disinfectant instead of a glucose solution by the nurse, which resulted in acute intoxication followed by the death of the pregnant woman and the fetus.

[Guidelines for antiretroviral therapy of human immunodeficiency virus (HIV)-infected subjects in the Czech Republic]. / Komplexní postup antiretrovirové lécby osob infikovaných virem lidské imunodeficience (HIV) v Ceské republice.

HIV infection remains an incurable disease because of the impossibility to eradicate the HIV from the organism. However, the combination antiretroviral therapy (cART) is able to efficiently limit HIV replication and slow down progression of immunodeficiency and thus prolong and improve the quality of HIV+ patients? lives. In HIV(+) pregnant women, the antiretroviral therapy substantially reduces the risk of vertical transmission of the infection. According to present knowledge, the cART is indicated mainly in symptomatic patients with stage B or C diseases and for vertical transmission and postexposure prophylaxis; less clear is the indication of cART for treating acute HIV infection and in asymptomatic patients. Various guidelines for the use of antiretroviral agents issued worldwide, e.g. in the USA, Europe or by the WHO, are not completely identical. The authors present a draft of recommendations for the use of antiretroviral agents in the Czech Republic based on the above-mentioned guidelines as well as on their own experience with taking care of HIV/AIDS patients.

[Prophylaxis of the vertical transmission of the human immunodeficiency virus (HIV) in the Czech Republic]. / Profylaxe vertikálního prenosu viru lidské imunodeficience (HIV) v Ceské republice.

GOAL: To assess the efficacy of prophylactic measures intended to reduce vertical transmission of HIV infection and to evaluate the predictive value of demographic, immunological and virological factors for determination of the risk of HIV transmission to fetus and neonate. METHODS: 56 pregnancies of 53 HIV-positive women were included in this retrospective study. The women have been in the care of the AIDS Centre of the Dpt. of Infectious Diseases, University Hospital Na Bulovce over the past 15 years. Cellular immunity tests and HIV RNA viral load in all subjects were regularly determined. In line with our present knowledge and the patients compliance, we introduced prophylactic measures, which included the administration of antiretroviral agents, delivery by Caesarean section and breast-feeding avoidance. 58 children were born from these pregnancies and repeated blood tests were performed to detect the presence or absence of HIV RNA. RESULTS: 58 infants were born from 56 pregnancies-and 3 of them (5.17%) were infected with HIV. In two mothers, we diagnosed HIV positivity few hours before delivery; another woman was diagnosed early, however, the failure of treatment was due to her insufficient compliance. In addition, premature amniorrhoea was present in two women. Moreover, one of them presented with untreated syphilis. The other 55 children stayed uninfected. In six of them, however, prophylactic measures were not fully followed, mainly because the patients disregarded them. CONCLUSIONS: The study fully confirms the high efficacy of prophylactic measures, which substantially reduce the risk of vertical transmission of HIV infection. Routine blood tests are necessary in all pregnant women by law. The failure of prophylaxis is in most cases due to inadequate compliance with the treatment. However, we cannot rule out the possible resistance to antiretroviral agents.

Unusual binding mode of an HIV-1 protease inhibitor explains its potency against multi-drug-resistant virus strains.

Protease inhibitors (PIs) are an important class of drugs for the treatment of HIV infection. However, in the course of treatment, resistant viral variants with reduced sensitivity to PIs often emerge and become a major obstacle to successful control of viral load. On the basis of a compound equipotently inhibiting HIV-1 and 2 proteases (PR), we have designed a pseudopeptide inhibitor, QF34, that efficiently inhibits a wide variety of PR variants. In order to analyze the potency of the inhibitor, we constructed PR species harboring the typical (signature) mutations that confer resistance to commercially available PIs. Kinetic analyses showed that these mutated PRs were inhibited up to 1,000-fold less efficiently by the clinically approved PIs. In contrast, all PR species were effectively inhibited by QF34. In a clinical study, we have monitored 30 HIV-positive patients in the Czech Republic undergoing highly active antiretroviral therapy, and have identified highly PI resistant variants. Kinetic analyses revealed that QF34 retained its subnanomolar potency against multi-drug resistant PR variants. X-ray crystallographic analysis and molecular modeling experiments explained the wide specificity of QF34: this inhibitor binds to the PR in an unusual manner, thus avoiding contact sites that are mutated upon resistance development, and the unusual binding mode and consequently the binding energy is therefore preserved in the complex with a resistant variant. These results suggest a promising route for the design of second-generation PIs that are active against a variety of resistant PR variants.

Response of HIV positive patients to the long-term salvage therapy by lopinavir/ritonavir.

BACKGROUND: The cohort of 19 patients on LPV/r salvage regimen was followed for the period of up to 37.5 months. Patient's virologic response was evaluated with regard to the various baseline characteristics. RESULTS: A 73.7% of patients (14 out of 19) achieved viral suppression during the first three months of treatment, either complete (47.4%) or partial (26.3%). This effect was only transient in five cases (virologic rebound emerged after 9 months of treatment on average) and in nine cases the treatment was successful in the long-term analysis (HIV RNA plasma level still undetectable at 31st month of the therapy on average with maximum of 36 months). We analyzed the link between the virologic response and possible predictive factors of treatment efficiency, such as lopinavir mutation score, various individual mutations, previous PI exposure, etc. We also describe changes in the PR sequence associated with poor response to the salvage therapy to LPV/r. CONCLUSIONS: The results of LPV/r salvage therapy were encouraging. About 47% of patients from our study achieved stable suppression of viral replication for 31 months on average. LPV/r proved to be potent inhibitor despite unfavourable prognosis.

[Cells of the immune system in the pathogenesis of HIV infections.].

For more than twenty years comprehensive research has been focusing on the pathogenesis of HIV infections. In this multifactorial complex a crucial role is played, in addition to virological aspects, by the mechanisms of the host's immune system. The paper focuses primarily on the role of T lymphocytes and of antigen - presenting cells - macrophages and dendritic cells - in HIV infections. It also discusses the control of HIV infections using neutralizing antibodies and HIV-1 specific CD8+ cytotoxic T lymphocytes and CD4+ T lymphocytes.

[Prevalence of GBV-C/HGV (HGV) in HIV-infected patients and potential influence of co-infection on the course of the disease.].

AIM OF THE STUDY: Assessment of prevalence of GBV-C/HGV infection in HIV-infected patients and evaluating of a possible influence of GBV-C/HGV on the course of HIV infection by assessment of immunological and virological markers of progression of HIV infection. MATERIALS AND METHODS: We have investigated serum samples of 273 HIV- infected patients from AIDS Center of the University Hospital Na Bulovce Prague during 2002-2003. Our target was to assess the presence of markers of GBV-C/HGV infection by semiquantitative HGV RNA evaluation and anti-E2 antibodies by ELISA testing. 271 of serum samples were tested for HGV RNA and 269 samples were tested for anti-E2 antibodies. HIV viral load and CD4 count were tested concurrently. We used Spearmans test to rule out the dependency of CD4 count and HIV viral load on HGV infection. RESULTS: 89 (33,3 %) of patients were positive in HGV RNA test and 101 (38,5 %) of patients were positive in anti E2-ELISA. No statistically significant effect of GBV-C/HGV infection was observed on CD4 count and HIV viral load in our cohort of patients. CONCLUSIONS: The effect of GBV-C/HGV infection on predictive laboratory markers of HIV infection was not confirmed in our study. Further investigations regarding this subject seem to be necessary.

[The issue of HIV/AIDS--present overview and perspectives]. / Problematika HIV / AIDS - ohlédnutí a perspektivy.

Many outstanding discoveries accompany the existence of HIV infections. International teams of scientists cooperate and in their work they use a great many methods from haematology and immunology to methods of molecular biology that pass into methods of molecular virology. These, in turn, open new prospects for methods of molecular epidemiology. The development of sensitive and specific antibody tests made possible the testing of blood intended for transfusions and helped map the epidemilogical situation, describe the clinical course of the disease and, in particular, diagnose the asymptomaic stage of infection. Gradually, the jigsaw puzzle of the pathogenesis of HIV infections was completed. The tremendous advances in antiretrovirus treament meant a turning point in the mortality and morbidity of HIV infection. Clinical trials of therapeutic vaccines are in progress. Yet despite all the advances and prevention efforts the HIV/AIDS pandemic continues.

Determination of muscarine in human urine by electrospray liquid chromatographic-mass spectrometric.

A liquid chromatography-mass spectrometry based method for determination of muscarine in human urine was developed and validated. The method involved a solid phase extraction of muscarine from urine using Strata X-CW column. Separation of muscarine was achieved within 16.0 min on a reversed phase Gemini C18 analytical column (150 mm × 2.0mm i.d., 5 µm) with a mobile phase consisted of aqueous 8 mmol/L heptafluorobutyric acid and acetonitrile in a gradient mode. Mass spectrometric detection was performed at m/z 174 and m/z 216 for muscarine and acetylmuscarine (internal standard), respectively. The linearity was satisfactory with a coefficient of determination (R(2)) 0.9993 at concentration range from 0.3 ng/mL to 2.0 µg/mL, LOD and LOQ for muscarine was 0.09 ng/mL and 0.3 ng/mL, respectively. The found out recoveries of muscarine were 96% or 95% for concentration 0.3 ng/mL and 0.2 µg/mL or 2.0 µg/mL, respectively. The precision in the intra-assay-study varied from 0.48% to 1.39% and in the inter-assay-study from 2.39% to 5.49%. The accuracy ranged from -3.3% to -6%. The validation results demonstrated that the method fulfilled satisfactory requirements for precision and accuracy across the calibration curve. The applicability of the method has been demonstrated by analyzing clinical urine samples. The method offers the fast objective identification of intoxication by muscarine and can become a routine screening alternative to more difficult microscopic examination of spores in the gastric content in clinical practice.

Effect of ethanol and hydrogen peroxide on poly(3-hydroxybutyrate) biosynthetic pathway in Cupriavidus necator H16.

Exposition of Cupriavidus necator to ethanol or hydrogen peroxide at the beginning of the stationary phase increases poly(3-hydroxybutyrate) (PHB) yields about 30%. Hydrogen peroxide enhances activity of pentose phosphate pathway that probably consequently increases intracellular ratio NADPH/NADP(+). This effect leads to stimulation of the flux of acetyl-CoA into PHB biosynthetic pathway and to an increase of enzymatic activities of ß-ketothiolase and acetoacetyl-CoA reductase while activity of PHB synthase remains uninfluenced. During ethanol metabolisation, in which alcohol dehydrogenase is involved, acetyl-CoA and reduced coenzymes NAD(P)H are formed. These metabolites could again slightly inhibit TCA cycle while flux of acetyl-CoA into PHB biosynthetic pathway is likely to be supported. As a consequence of TCA cycle inhibition also less free CoA is formed. Similarly with hydrogen peroxide, activities of ß-ketothiolase and acetoacetyl-CoA reductase are increased which results in over-production of PHB. Molecular weight of PHB produced under stress conditions was significantly higher as compared to control cultivation. Particular molecular weight values were dependent on stress factor concentrations. This could indicate some interconnection among activities of ß-ketothiolase, acetoacetyl-CoA reductase and PHB molecular weight control in vivo.