Multi-locus sequence typing and phylogenetics of Cryptococcus neoformans AD hybrids.

Hybrid AD strains of the human pathogenic Cryptococcus neoformans species complex have been reported from many parts of the world. However, their origin, diversity, and evolution are incompletely understood. In this study, we analyzed 102 AD hybrid strains representing 21 countries on five continents. For each strain, we obtained its mating type and its allelic sequences at each of the seven loci that have been used for genotyping haploid serotypes A and D strains of the species complex by the Cryptococcus research community. Our results showed that most AD hybrids exhibited loss of heterozygosity at one or more of the seven analyzed loci. Phylogenetic and population genetic analyses of the allelic sequences revealed multiple origins of the hybrids within each continent, dating back to one million years ago in Africa and up to the present in other continents. We found evidence for clonal reproduction and long-distance dispersal of these hybrids in nature. Comparisons with the global haploid serotypes A and D strains identified new alleles and new haploid multi-locus genotypes in AD hybrids, consistent with the presence of yet-to-be discovered genetic diversity in haploid populations of this species complex in nature. Together, our results indicate that AD hybrids can be effectively genotyped using the same multi-locus sequencing type approach as that established for serotypes A and D strains. Our comparisons of the AD hybrids among each other as well as with the global haploid serotypes A and D strains revealed novel genetic diversity as well as evidence for multiple origins and dynamic evolution of these hybrids in nature.

Association of CAST gene polymorphism with carcass value and meat quality in two synthetic lines of sheep.

The aim of study was to estimate genotype frequencies of the polymorphism in the intron 12 of CAST gene of growing lambs, and to evaluate associations between the CAST genotype and carcass and meat traits. A total number of 317 rams representing the synthetic BCP (163) and SCP (154) meaty lines were genotyped by the RFLP/MspI/NcoI/Hin6I. Three genotypes including "aa", "ae" and "ac" with most frequencies of 0.48, 0.27 and 0.19, were observed in whole population. Lambs with the "aa" CAST genotype showed the highest muscle percentage and the lowest fat percentage in the hind legs. On the other hand, the loins of rams with "ac" genotype had the highest intramuscular fat content, indicative of beneficial health-related properties and technological usefulness. However, differences between CAST genotype stated for the sensory properties, texture and thawing and cooking losses of the meat were insignificant. Breeding efforts aimed at the improvement of lamb meat quality should prioritize the selection of the "ac" genotype.

Adhesion of Malassezia pachydermatis of different growth type to canine epithelial cells.

A total of 100 Malassezia pachydermatis strains recovered from skin and mucosal membranes of dogs were evaluated for their adhesive properties. Two types of growth, related to colony morphology on Sabouraud agar, were observed (type I and II). The mean number of fungal cells attaching to canine buccal epithelial cells was found to be 17. The number of adhered cells was greater (statistically significant at the level of p < 0.01) in strains belonging to the type I.

Distributions of (137)Cs and (210)Pb in moss collected from Belarus and Slovakia.

In the present work, moss samples collected in Slovakia and Belarus were assayed with respect to gamma-emitting radionuclides. The results for (137)Cs and (210)Pb are discussed. Moss was used for the first time in Belarus, as a biological indicator of radioactive environmental pollution in consequence of the Chernobyl accident in 1986. In Belarus, the maximum activity of (137)Cs was observed in the Gomel region near Mazyr (6830 Bq/kg) and the minimum activity in the Vitebsyevsk Region near Luzhki-Yazno (5 Bq/kg). "Hot spots" were also observed near the towns Borisow and Yuratsishki. The results of measurements of (137)Cs in moss samples collected in 2000, 2006 and 2009 in the same localities of Slovakia are presented and compared with the results of air monitoring of (137)Cs carried out in Slovakia from 1977 until 2010. Measurements of the (210)Pb concentration in moss samples collected over the territory of Slovakia showed, that the median value exceed 2.3 times median value of (210)Pb obtained for Belarus moss. For that reason, the inhalation dose for man from (210)Pb and (137)Cs in Slovakia is more than twice as high as in Belarus, in spite of the initially very high (137)Cs exposure in the latter country.

Sirolimus and mycophenolate mofetil as GVHD prophylaxis in myeloablative, matched-related donor hematopoietic cell transplantation.

We investigated sirolimus and mycophenolate mofetil (MMF) as GVHD prophylaxis in patients with advanced hematological malignancies receiving myeloablative hematopoietic cell transplantation (HCT) from HLA-identical sibling donors. On the basis of pre-study stopping rules, the trial was closed to accrual after enrollment of 11 adult patients. In all, 7 of the 11 patients received BU-containing preparative regimens. Sirolimus was discontinued in three patients because of the toxicity-related events of severe sinusoidal obstructive syndrome, portal vein thrombosis, altered mental status and in one patient because of the risk of poor wound healing. In all, 6 of the 11 patients developed grade II-IV acute GVHD (AGVHD) a median of 15.5 days post HCT. Two of three patients with grade IV AGVHD had sirolimus discontinued by 9 days post HCT. All patients responded to AGVHD therapy without GVHD-related deaths. There were two non-relapse- and two relapse-related deaths. At a median follow-up of 38 months (2-47 months), 7 of 11 patients were alive without disease. MMF and sirolimus GVHD prophylaxis did not reduce the risk of AGVHD, however, there were no GVHD-related deaths. The severe toxicities in the patients receiving the BU-containing preparative regimens limited the continued use of sirolimus and MMF for the prevention of AGVHD.

Chemical composition and inherent properties of offal from calves maintained under two production systems.

The aim was to evaluate the influence of organ and calf type on the chemical composition, mineral concentration, fatty acid profile and inherent properties in offal. Organ type influenced (p<0.01) all evaluated characteristics, whereas calf type to a lesser extent affected mineral content. The highest contents of K, Zn and Fe were found in liver. The heart of veal calves contained the highest contents of Mg and Mn. In addition, the most favorable fatty acid profile was also found in hearts, by reason of the lowest SFA and highest PUFA percentage and h/H ratio. Significantly higher values of PUFA percentage, PUFA/SFA ratio, and content of CLA were found in organs of suckler beef, which is associated with the beneficial effects of grazing on the fatty acid profile. Results could help to increase the consumption of calves' offal and its utilization in meat processing.

Blood haematological profile and meat colour of calves slaughtered in summer and autumn season.

The purpose of this study was to compare the haematological profile and meat colour of calves slaughtered in summer and autumn. The material covered 42 calves chosen randomly i.e. 22 slaughtered in summer (June-August) and 20 in autumn (October-December). Haematological analyses included haematocrit (HCT), haemoglobin level (HGB), red blood cells (RBC), white blood cells (WBC) and platelets (PLT). The morphology of the erythrocytes and platelets, and differentiation of leucocytes were examined. The colour of meat was evaluated instrumentally by Minolta CIE L*a*b* and haematin pigment content was determined. The blood haemoglobin content in calves in the compared seasons was similar (11.3 g/L) and found within a normal range. Blood of calves from the autumn season showed higher HCT, RBC, and MCV values with concurrent lower MCH and MCHC values in comparison to the summer season. Blood of calves slaughtered in summer showed a higher content of WBC and a significantly higher percentage of lymphocytes as against blood of calves from the autumn. Meat of calves from the summer season was paler (higher L* value), and had a significantly (p < or = 0.05) higher proportion of yellowness (b*). A brighter colour of meat from calves presented for slaughter in the summer season was noted along with a lower content of haematin pigments. Significant correlations were found between haematological variables (HGB, HCT and RBC, particularly) and haematin pigment content and meat lightness (L*) and redness (a*).

Providencia alcalifaciens as the presumptive cause of diarrhoea in dog.

A case of diarrhoea in a four-month-old golden retriever is described. On the basis of anamnesis and bacteriological examination, the diagnosis of bacterial enteritis due to Providencia alcalifaciens was reached. Although there are contradictory opinions about the role of this organism as the enteric pathogen, it seems that Providencia alcalifaciens should also be taken into consideration in the routine bacteriological diagnostics of diarrhoea in dogs.

Caspofungin treatment in severely ill, immunocompromised patients: a case-documentation study of 118 patients.

BACKGROUND: Caspofungin has shown efficacy in empirical antifungal therapy in neutropenic patients, refractory invasive Aspergillus infections and invasive candidiasis. Here we report the currently largest series of patients treated with caspofungin outside clinical trials. METHODS: Centres in Germany that were known to treat patients with invasive fungal infections were asked to fill out detailed questionnaires for all patients treated with caspofungin. No effort was made to influence the decision to use caspofungin. RESULTS: A total of 118 patients were evaluable (median age 48 years, interquartile range 38-58), out of which 41 (35%) suffered from acute leukaemia, 31 (26%) had allogeneic stem cell transplants, 16 (14%) lymphoma or myeloma, 8 (7%) autologous stem cell transplants and 22 (19%) other causes for immunosuppression. One hundred and six patients were evaluable for efficacy out of which 68 (64%) patients achieved a complete or partial remission. A total of 81 out of 115 (70%) patients were alive 30 days after the end of caspofungin therapy. Response rates were similar in proven (20/32, 63%) and probable (27/46, 59%) infections, in neutropenic patients (41/55, 75%) and in patients who were (44/70, 63%) or were not (24/36, 67%) refractory to antifungal pre-treatment. The response rate in mechanically ventilated patients was 29% (7/24). Caspofungin was well tolerated, even in 14 patients, who were concomitantly treated with ciclosporin A, no drug-related elevations of bilirubin, alanine aminotransferase or creatinine were found. CONCLUSIONS: This open case study of severely ill patients with invasive fungal infections demonstrates both excellent efficacy and very low toxicity of caspofungin.

Sequential monitoring of chimerism and detection of minimal residual disease after allogeneic blood stem cell transplantation (BSCT) using multiplex PCR amplification of short tandem repeat-markers.

Sequential analysis of chimerism after allogeneic blood stem cell transplantation (BSCT) has been shown to be predictive for graft failure and relapse. We have explored the impact of a novel approach for the quantitative determination of chimerism using a commercial PCR assay with multiplex amplification of nine STR-loci and fluorescence detection. The feasibility was studied in 121 patients transplanted from related or unrelated donors. Follow-up investigation was performed in 88 patients. Twenty-eight of these patients had received a transplantation after dose-reduced conditioning therapy. Results were compared to data obtained by FISH analysis in a subgroup of patients receiving grafts from sex-mismatched donors. The analysis was possible in all patients, the median number of informative alleles was 4 (range 1-8) compared to 7 (range 1-9) in the related and unrelated situation, respectively. A good correlation was seen in 84 samples from 14 patients analyzed in parallel with STR-PCR and FISH. Decreasing values of donor chimerism were detected prior to or concomitantly with the occurrence of graft failure and relapse of disease in all patients investigated prospectively. Using FACS-sorted material, eg peripheral blood CD34+ cells, the assay permitted the detection of residual recipient cells with high sensitivity (down to one CD34+ Kasumi cell in 40,000 normal WBC). Evaluation of the inter-laboratory reproducibility revealed that in 20 samples analyzed in three different centers, the median coefficient of variation was 2.1% (range 0.7-9.6%). Taken together, the results support the use of the test as a valuable tool in the follow-up of patients undergoing allogeneic BSCT. In cases lacking PCR-detectable disease-specific gene products, this assay may represent an alternative to recently established real-time PCR methods.

Rapid quantification of mixed chimerism using multiplex amplification of short tandem repeat markers and fluorescence detection.

Monitoring the engraftment of donor cells after allogeneic blood stem cell transplantation (BSCT) may be important for the early diagnosis of graft failure or relapse of disease. Several techniques have been reported for this purpose. PCR-based assays analyzing polymorphic short tandem repeat (STR) markers are attractive because they are sensitive and can be performed rapidly. The intent of the present study was to test a novel approach for the quantification of mixed chimerism using a commercial multiplex STR assay with fluorescence-based detection for forensic purposes. The feasibility of this assay and the accuracy of quantitative results was tested using serial cell mixtures of unrelated individuals. Sample preparation was optimized to obtain information from minute amounts of starting material, eg from patients with aplasia or from sorted cell populations. Using the STR-PCR, discrimination between donor and recipient was possible in all patients analyzed (n = 25). Cell dilution experiments showed a linear correlation between the cell numbers added and the proportions found, with the limit of detection for a minor cell population being 5%. Comparison of values obtained with standard FISH analysis in patients transplanted from sex-mismatched donors showed an excellent correlation with the STR-PCR results. Taken together, this procedure allows the rapid, versatile and accurate quantification of mixed chimerism, even with minuscule numbers of cells.

A cerebrospinal fluid turnover in Parkinson's disease.

In 30 patients with Parkinson's disease (PD) CT scan, isotopic cisternography and neuropsychological examination were performed to test possible relationship between cerebrospinal fluid (CSF) turnover, intellectual impairment and brain atrophy. Pathological cisternography was found in one third of the patients and it was positively correlated with the Columbia Rating Scale. Cerebral atrophy on CT was found more frequently in patients with earlier onset of PD. Combined pathological pattern on CT scan and on cisternography was correlated with intellectual decline. We conclude from this study, that in PD intellectual deterioration and brain atrophy separately are not connected with abnormal CSF turnover.

[Disorders and difficulties in the psychosocial adaptation of epileptics]. / Zaburzenia i trudnosci w przystosowaniu psychospolecznym u chorych z padaczka.

Disturbances of and difficulties in psychosocial adaptation were analysed in 63 epileptics (with generalized convulsions and partial complex seizures) and in 31 subjects without diseases of the central nervous system (control group). The disturbances of adaptation were not more frequent in epileptics than in controls. On the other hand, it was observed that patients with partial complex seizures constituted a group non-homologous with respect to the degree of difficulties in functioning (disturbances and difficulties) which was probably due to their greater dependence on the conditions of realization of their psychosocial needs in relation to healthy subjects and patients with generalized seizures.

[Perseverance tendency in patients with generalized convulsive seizures and with partial seizures with complex symptoms]. / Tendencja do perseweracji u chorych z uogólnionymi napadami drgawkowymi oraz z napadami czesciowymi o symptomatologii zlozonej.

The intensity of perseveration was tested in 70 patients with generalized convulsive seizures and with partial seizures of complex symptomatology, and in 37 subjects without central nervous system diseases (controls). The results suggest that the tendency for perseveration is a feature occurring not only in epilepsy but also in healthy subjects (probably of constitutional character); the disease process responsible for epileptic seizures can intensity this feature in a degree depending on its duration.

Intelligence level of patients with the Duchenne type of progressive muscular dystrophy (pmd-d).

The I.Q. of 129 patients with PMD-D and 27 patients suffering from Werdnig-Hoffmann disease were estimated. Among the patients with PMD-D there was one group without any complicating factors and 3 other groups with additional factors that might influence the intelligence level. Comparing mean values and distribution of I.Q. for all these groups, one can conclude that, besides additional unfavourable pathological and environmental factors in all cases, PMD-D itself causes a small decrease of the I.Q. by about 1 SD. The frequent changes of the EEG record in these patients could reflect involvement of the CNS by the pathological process.