Changes in antioxidant,inflammatory and metabolic markers during 1 week cultivation of human skin explants

Human skin explant (HSE) seems to be a useful model for dermatological/cosmetic testing. HSE prepared from donor superfluous skin from plastic surgery operations is cheap and easily obtainable compared to reconstructed models. The HSE use, however, may be limited by the degeneration processes during cultivation. The aim was to monitor changes in metabolic activity and selected apoptotic, inflammatory and antioxidant parameters during 7 day cultivation. The significant changes were found in the superoxide dismutase‐2 level from day 5, glutathione S‐reductase level from day 6, metabolic activity and fibulin‐5 level from day 4, cyclooxygenase‐2, interleukin‐6 and interleukin‐10 from day 1 to 2. Other selected markers (lipid peroxidation products and glutathione level, glutathione S‐transferase, catalase, superoxide dismutase and glutathione S‐reductase activity, glutathione peroxidase and glutathione S‐reductase levels) were not modified significantly due to high inter‐individual variability of skin donors. The HSE microstructure as well as cytokeratin‐10 and proliferation marker Ki67 expression was also only minimally affected during cultivation. Collectively, the results demonstrate that HSE represents a good model for short‐term studies focused on the physical and chemical agent toxicity, protective potential of compounds or metabolic biotransformation. However, reduced metabolic activity, increased inflammation and the high inter‐individual variability and sensitivity of donors have to be taken into consideration.     

Minimal residual disease detectable by quantitative assessment of WT1 gene before allogeneic stem cell transplantation in patients in first remission of acute myeloid leukemia has an impact on their future prognosis

Overall 42 patients (pts) transplanted in hematological CR1 were retrospectively analyzed. Median follow‐up was 15 months (range 2–77). The expression of WT1 gene was measured according to the European Leukaemia Net recommendations. At the time of allogeneic stem cell transplantation (allo‐SCT) 29 pts were WT1‐negative and 13 pts were WT1‐positive. In the univariate analysis, significantly better results were observed in the group of WT1 neg in terms of progression‐free survival (in three yr 77% vs. 27%, p = 0.001). In multivariate analysis, the only significant feature in terms of better OS was WT1 negativity (p = 0.029). Our results show that minimal residual disease status measured by quantitative assessment of WT1 gene in acute myeloid leukemia pts in CR1 significantly affects their future prognosis after allo‐SCT.     

Reversible arrhythmias observed in patients treated with recombinant alpha2 interferon

Summary We have treated 25 patients, 7 with breast cancer and 18 with non-Hodgkin's lymphoma, with recombinant alpha₂ interferon. In 5 patients we observed cardiac arrhythmias that were unexpected and required treatment. No deaths have occured that we can attribute to interferon, though 1 patient had to be resuscitated. Age, prior cardiac disease, prior treatment with doxorubicin, and interferon dose appear to be predisposing factors for this toxicity.Supported by the Schering Corporation and Harper-Grace Hospitals     

Subtyping Cryptosporidium ubiquitum,a Zoonotic Pathogen Emerging in Humans

Cryptosporidium ubiquitum is an emerging zoonotic pathogen. In the past, it was not possible to identify an association between cases of human and animal infection. We conducted a genomic survey of the species, developed a subtyping tool targeting the 60-kDa glycoprotein (gp60) gene, and identified 6 subtype families (XIIa–XIIf) of C. ubiquitum. Host adaptation was apparent at the gp60 locus; subtype XIIa was found in ruminants worldwide, subtype families XIIb–XIId were found in rodents in the United States, and XIIe and XIIf were found in rodents in the Slovak Republic. Humans in the United States were infected with isolates of subtypes XIIb–XIId, whereas those in other areas were infected primarily with subtype XIIa isolates. In addition, subtype families XIIb and XIId were detected in drinking source water in the United States. Contact with C. ubiquitum–infected sheep and drinking water contaminated by infected wildlife could be sources of human infections.Key words: Cryptosporidiosis, Cryptosporidium, zoonoses, epidemiology, molecular typing,whole genome sequencing, genomics, ruminants, rodents, horse, raccoon, sifaka, parasites, humansCryptosporidium infection is a leading cause of diarrhea in humans (1). Five Cryptosporidium species—C. hominis, C. parvum, C. meleagridis, C. felis, and C. canis—are responsible for most cases of cryptosporidiosis in humans. Among them, C. hominis and C. parvum are the most common etiologic agents, and the latter is responsible for most zoonotic infections (2). In recent years, C. ubiquitum, previously known as the cervine genotype, has been emerging as another major zoonotic species that infects persons. It has been found in humans worldwide, primarily in industrialized nations (3–11). In the United Kingdom, more human cases of cryptosporidiosis have been attributed to C. ubiquitum than to C. canis (9).C. ubiquitum is of public health concern because of its wide geographic distribution and broad host range. Of all Cryptosporidium spp. identified by molecular diagnostic tools, it infects the greatest variety of host species (12). C. ubiquitum has been commonly detected in domestic and wild ruminants (sheep, goats, mouflon sheep, blesboks, nyalas, white-tailed deer, Père David’s deer, sika deer, ibexes, buffalos, and yaks), rodents (squirrels, chipmunks, woodchucks, beavers, porcupines, deer mice, house mice, and gerbils), carnivores (raccoons), and primates (lemurs and humans) (12–16). It has also been found in drinking source water, storm water runoff, stream sediment, and wastewater in various geographic locations (17–22).Thus far, showing an association between human and animal cases of C. ubiquitum infection has not been possible because of the lack of suitable genetic markers for subtyping. For C. parvum, C. hominis, and some genetically related species, the most commonly used marker for subtyping is the 60-kDa glycoprotein gene (gp60, also called gp40/15). Sequence analysis of the gp60 gene has been used in studies of the genetic diversity, host adaptation, infection sources, and transmission dynamics of these Cryptosporidium spp. (2). However, it has been suggested that a single locus, such as gp60, is not a reliable marker of C. parvum and C. hominis population structure because genetic recombination may occur (23).Because C. ubiquitum is genetically distant from C. hominis and C. parvum, its homologue of the gp60 gene has thus far not been identified (24). In this study, we identified the gp60 gene of C. ubiquitum by whole-genome sequencing and used it to develop a subtyping technique to characterize specimens from humans, various animals, and drinking source water.     

Operative treatment of infective endocarditis in children

A retrospective review of 11 children, aged 2 months to 15 years, demonstrates the feasibility of surgical treatment for infective endocarditis in childhood. Except for one case of perinatal infection, in all instances the infective endocarditis was a complication of a congenital heart defect. As medical treatment was not successful, surgery was indicated. Debridement of infected tissue and repair of the congenital heart defect was performed. There were no septic complications postoperatively although 8 patients were operated upon during the active stage of infection. One 2-month-old child did not survive excision of an infected tricuspid valve. The follow-up period of 8 years to 5 months (median 39 months) showed a good haemodynamic result (NYHA class I) in the remaining 10 patients. This included 4 patients with prosthetic valves.     

Unapparent microsporidial infection among immunocompetent humans in the Czech Republic

In the present population-based study, we determined the prevalences of the most common human-pathogenic microsporidia, Encephalitozoon spp. and Enterocytozoon bieneusi, in asymptomatic healthy people living in the Czech Republic. A total of 382 males and females (ages, 1 to 84 years) living in the Czech Republic, of whom 265 were Czech nationals and 117 were foreign students, were included in a study testing for the presence of microsporidia by use of coprology and molecular methods. Single-species infections with Enterocytozoon bieneusi or an Encephalitozoon sp. were detected for 9 and 136 individuals, respectively. Moreover, coinfections were detected for 14 individuals. Four genotypes of 3 human-pathogenic Encephalitozoon spp. and 7 E. bieneusi genotypes, including 3 novel genotypes, were detected. Some of these were reported in humans for the first time. The highest prevalence was recorded for individuals older than 50 years and for loose, unformed stool samples. These findings clearly show that exposure to microsporidia is common among immunocompetent people and that microsporidiosis is not linked to any clinical manifestation in healthy populations.