Flow cytometric analysis of CD55 and CD59 expression on blood cells in paroxysmal nocturnal haemoglobinuria

PNH is a rare clonal disorder of hematopoietic stem cells, therefore all blood cells lineages are involved. The main feature is an increased sensitivity of erythrocytes to complement-mediated cell lysis due to deficiency of membrane-bound GPI (glycosylphosphatidylinositol)-anchored proteins which normally function as inhibitors of reactive hemolysis. In the present study, we performed flow cytometric analysis using monoclonal antibodies against CD55 and CD59 for the detection of PNH-type clone in the blood of 50 patients (28 females and 22 males, age range 7-67 yrs). In one patient only we found a large population (95%) of granulocytes with decreased expression of both CD55 and CD59 molecules (type I PNH) and in two others with partial loss of CD55 expression (type II PNH). The expression was determined chiefly on granulocytes which in the control group showed reliable and high expression of CD55 and CD59.     

A Study of Glutathione <Emphasis Type="Italic">S</Emphasis>-transferase pi Expression in Central Nervous System of Subjects with Amyotrophic Lateral Sclerosis Using RNA Extraction from Formalin-Fixed,Paraffin-Embedded Material

The expression of glutathione S-transferase pi (GST pi), an enzyme responsible for inactivation of a large variety of toxic compounds was studied in spinal cord, motor and sensory brain cortex obtained from patients who died in the course of amyotrophic lateral sclerosis (ALS). The studies were performed on formalin-fixed, paraffin-embedded (FFPE) and freshly frozen tissues. The method of RNA isolation from FFPE was modified. A significant decrease of GST pi-mRNA expression was found in cervical spinal cord and motor brain cortex of ALS subjects comparing to analogue control tissues (P < 0.01), as well as in motor cortex of ALS subjects comparing to their sensory cortex (P < 0.05). In spinal cords the decrease in GST pi-mRNA expression was accompanied by a decrease of GST pi protein level. Results indicated lowered GST pi expression on both mRNA and protein levels in the regions of nervous system affected by ALS. The non-properly inactivated by GST toxic electrophiles and organic peroxides may thus contribute to motor neurons damage.     

Age effect of broiler breeders on fertility and sperm penetration of the perivitelline layer of the ovum

The objective of this study was to determine the age effect of a broiler breeder flock on duration of fertility and number of spermatozoa penetrating the perivitelline layer overlying the germinal disc (SP/mm(2) GDIPVL). Moreover, in the second half of the flock's reproductive life, the effect of using ejaculates of young roosters (CA2) in artificial insemination (AI) on the above parameters of fertility was estimated. The commercial flock of broiler breeder hens (n = 100) was inseminated six times from 31 to 62 weeks of age. Additional inseminations, with ejaculates of roosters aged 31 and 36 weeks (CA2), were performed at 56 and 62 weeks of age. AI was performed during two consecutive days (D0 and D1) with an insemination dose of 125 x 10(6) spermatozoa/0.06 ml containing pooled ejaculates. The following parameters were studied: the effective and maximum duration of fertility (De and Dm), percent of fertility on different days after AI (FD10, FD15 and FD20), indices of duration of sperm penetration (DSP, SP < or = 3/GDIPVL), SP/mm(2) GDIPVL in eggs laid on successive days after insemination of hens at different age, and correlations between some fertility indices. Both for De and Dm, the highest values were noted after AI of the layers at 36 weeks of age (14.8 +/- 0.49 and 17.4 +/- 0.46 days, respectively), which were about 2 days longer than at 56 weeks. All fertility indices decreased gradually with age, starting from AI at peak egg production (31-36 weeks of age), while the use of ejaculates from CA2 did not help to increase them significantly. Correlation coefficients between SP/mm(2) GDIPVL and the other fertility indices were positive and highest for eggs laid on D3. It is concluded that high De values can be obtained from broiler breeders in adequate environmental and technological conditions of AI. It is suggested that the age-related decrease in fertility is more pronounced in females, in which the efficiency of sperm storage tubules decreases. The present fertility indices indicate the possibility of lengthening AI intervals, especially at peak egg production.     

HMGA proteins and their genes as a potential neoplastic biomarkers

HMGA proteins and their genes are described in this article. HMGA proteins reveal ability to bind DNA in AT-rich regions, which are characteristic for gene promoter sequences. This interaction lead to gene silencing or their overexpression. In normal tissue HMGA proteins level is low or even undetectable. During embriogenesis their level is increasing. High HMGA proteins level is characteristic for tumor phenotype of spontaneous and experimental malignant neoplasms. High HMGA proteins expression correlate with bad prognostic factors and with metastases formation. HMGA genes expression can be used as a marker of tumor progression. Present studies connected with tumor gene therapy based on HMGA proteins sythesis inhibition by the use of viral vectors containing gene encoding these proteins in antisence orientation, as well as a new potential anticancer drugs acting as crosslinkers between DNA and HMGA proteins suggest their usefulness as a targets in cancer therapy.     

Influence of hormonal therapy on growth rate and bone age progression in patients with Turner syndrome

The efficacy of growth promoting hormonal therapy is assessed on the basis of growth rate as well as bone age progression until the patients reach their final height. The aim of our study was to investigate which hormonal therapy influences in most appropriate way height velocity and bone age progression in patients with Turner syndrome (TS) and to establish the optimal age to initiate treatment. Patients were divided into five groups according to the type of hormonal therapy: 1) 11 patients treated with growth hormone (GH); 2) 18 patients treated with GH and oxandrolone (Ox); 3) 7 patients treated with GH, Ox and estrogens (E); 4) 6 patients treated with OX and E; and the control group (Group 0) of 62 untreated patients. The patients height was expressed in hSDS calculated on the basis of growth chart for patients with TS (hSDST). Bone age (BA) was assessed according to Greulich-Pyle method. Results: The mean values of deltahSDST in the first and second year of therapy in individual groups were significantly different. The difference resulted from significantly higher value of deltahSDST in group treated with GH+Ox. Analysis of regression between deltaCA and deltaBA revealed regression coefficients alpha of equation deltaBA= alpha x deltaCA: in group 0: 0.817; group GH: 1.233; group GH+Ox: 0.861; group GH+Ox+E: 0.997; group Ox+E: 1.141. There was significant difference between regression coefficients in studied groups. It resulted from significantly higher value of alpha in group treated with GH than in a group 0 and treated with GH+Ox. Only group treated with GH+Ox showed a significant negative correlation between baseline CA and deltaBA during the therapy. We can conclude that all regimens of hormonal therapy improved height in our patients but the highest increase of height during the therapy and the smallest progression of the bone age in the same time were observed in patients treated with GH+Ox.     

Metabolism of phenolic compounds in <Emphasis Type="Italic">Vitis riparia</Emphasis> seeds during stratification and during germination under optimal and low temperature stress conditions

We studied the alterations in phenolic compounds in grape seeds during their stratification and germination under optimal conditions (+25 °C) and at low temperature (+10 °C). Biological materials in the study were seeds of Vitis riparia. Phenolic compounds were extracted from defatted seeds using 80 % methanol or 80 % acetone. The content of total phenolics was determined with the Folin-Ciocalteau reagent, while the content of tannins was determined by vanillin assay and the protein (BSA) precipitation method. The RP-HPLC method was used to determine phenolic compounds (phenolic acids, catechins) in the extracts. High amounts of tannins, catechins, gallic acid and lesser amounts of p-coumaric acid were found in the seeds. The content of total phenolics in acetone extracts was higher than that obtained using methanol. The amounts of phenolic acids and tannins found in V. riparia seeds after stratification were much lower. It may confirm a possible role of these compounds in dormancy of V. riparia seeds. After 72 h of low temperature treatment, inhibition of grape root growth and biochemical changes in seeds were detected. The chilling stimulated increased accumulation of some phenolic compounds (free gallic acid and catechins) in the seeds. These substances can protect plants against some abiotic stressors.     

Application of Image Analysis Techniques in Activated Sludge Wastewater Treatment Processes

Image analytical techniques have been extensively developed to evaluate complex microbial aggregates such as sludge flocs and biofilms. This review covers the latest contributions concerning the application of image analysis to the activated sludge systems with respect to the most frequently used morphological parameters and relations between them and traditional wastewater treatment parameters. Recent developments have indicated that image analysis can be successfully used for the quantification of flocs and filamentous bacteria in the operating wastewater treatment plants, which enables prediction of bulking events and pinpoint flocs formation.