Purification and properties of phenylalanyl aminopeptidase synthesised by Pseudomonas sp.

Intracellular aminopeptidase synthesized by a soil strain of Pseudomonas sp. was purified 323-fold using the following procedure: saturation with ammonium sulfate, separation by preparative electrophoresis, anion-exchange chromatography and gel filtration chromatography. Molecular weight of the enzyme determined according to the latter method was 57 kDa.

The use of chosen serological diagnostic methods in Lyme disease in horses. Part II. Western blot.

In this investigation the Western blot test was treated as a method verifying results of the IFA, commercial ELISA and standardized ELISA tests (described in Part I). The verifying investigations were performed on 82 serum samples, which in the commercial ELISA were positive in 36 cases, dubious in 31 cases and negative in 15 cases as well as on 5 serum samples obtained from horses infected with Leptospira spp., which in the ELISA commercial were dubious (total of 87 sera samples).

The use of chosen serological diagnostic methods in Lyme disease in horses. Part I. Indirect immunofluorescence and enzyme-linked immunosorbent assay (ELISA).

The investigations aimed to establish the reliability of the chosen serological tests designed for the diagnosis of Lyme borreliosis in horses. The investigations were carried out in five Horse Breeding Centres (OHK). Statistical analysis methods were used to determine sample size for particular centres: Krasne (Kr)--49, Łack (Ł)--21, Walewice (W)--111, BogusŁawice (B)--17, Kozienice (K)--61.

Preconditioning with millimolar concentrations of vitamin C or N-acetylcysteine protects L6 muscle cells insulin-stimulated viability and DNA synthesis under oxidative stress.

The effect of reactive oxygen/nitrogen species (ROS/RNS)(hydrogen peroxide -- H(2)O(2), superoxide anion radical O(2)*- and hydroxyl radical *OH -- the reaction products of hypoxanthine/xanthine oxidase system), nitric oxide (NO* from sodium nitroprusside -- SNP), and peroxynitrite (ONOO(-) from 3-morpholinosydnonimine -- SIN-1) on insulin mitogenic effect was studied in L6 muscle cells after one day pretreatment with/or without antioxidants. ROS/RNS inhibited insulin-induced mitogenicity (DNA synthesis). Insulin (0.

Rats with a glucocorticoid-induced catabolic state show symptoms of oxidative stress and spleen atrophy: the effects of age and recovery.

In this study we wanted to determine whether changes in antioxidant profile could follow the catabolic effects of glucocorticoids. We also wanted to compare resistance to glucocorticoid overload in young and old rats. To address these questions, whole body catabolism was induced by the administration of dexamethasone (Dex) at either 2 mg/kg bodyweight/day to young (6 weeks old) or 0.

Regulation of the activity of intracellular alanylaminopeptidase synthesized by Pseudomonas sp.

Activity of purified alanylaminopeptidase of Pseudomonas sp. measured in the presence of the alanine derivative of 2-naphthoic acid (NA-Ala) is inhibited by 1,10-phenanthroline, EDTA, bestatin and amastatin; this finding supports the conclusion that this enzyme is a metallo-aminopeptidase. A decrease of its activity in the presence of iodoacetamide and its activation by thiols points to the significant role of -SH groups in the regulation of its activity.

Microsatellite polymorphism in locus OMHC1 (MHC Class I) in Polish Heath Sheep and Polish Lowland Sheep (Zelazna variety).

Our study aimed at comparative analysis of microsatellite polymorphism in locus OMHC1 (MHC Class I) in Polish Heath Sheep and Polish Lowland Sheep (Zelazna variety). The study was conducted on 100 ewes of each breed. We identified 13 alleles of the gene in Polish Heath Sheep and 9 in Polish Lowland Sheep.

Apoptosis during ectromelia orthopoxvirus infection is DEVDase dependent: in vitro and in vivo studies.

Ectromelia virus (EV), which causes mousepox, is a member of the orthopoxviruses that are defined as being able to suppress apoptosis. Caspase-3 is one of the key effector proteases which regulates the apoptotic cascade and which is responsible for DNA fragmentation observed during apoptosis. It is well known that viruses, especially poxviruses, can inhibit caspase activity.

Diets containing leguminous seeds influence chromium content in the rat femur bone.

Leguminous seeds contain a lot of antinutritional factors (ANFs) such as protease inhibitors, lectins and condensed tannins which can affect bioavailability of nutrients. Detrimental effect of protease inhibitors can be decreased by sulphur amino acids addition. Moreover, most of the leguminous ANFs are thermolabile.

CCK1 central receptor antagonist prevented the intestinal stress symptoms in sheep.

The aim of this study was to determine the influence of mechanically induced duodenal distension (DD) and lorglumide (CCK1 receptor antagonist) premedication on electrical activity of various parts of gastrointestinal (GI) tract and the blood plasma cortisol level in sheep. The influence of lorglumide on the unfavourable effects of duodenal distension (performed with a balloon filled with water--40 and 80 ml; DD40 and DD80) was investigated in this study. These effects in sheep were as follows: the atony of forestomachs and abomasum and the transitory stimulation of myoelectrical activity of small intestine and distal parts of large intestine.

Estrogen receptors in canine mammary gland tumours.

The aim of the present study was to establish the relationship between estrogen receptor expression in neoplastic parenchymal cells and the type of canine mammary gland tumour. The research material included mammary gland tumours obtained from 66 dogs. Paraffin sections were stained with hematoxylin-eosin and immunohistochemically with monoclonal antibodies and LSAB/Peroxidase/Universal Kit.

A useful protocol for in situ RT-PCR on plant tissues.

This study describes an effective method of in situ RT-PCR (RT-ISPCR) that was developed to localize gene expression in plant tissues. This RT-PCR technique was performed on sectioned tissues of female buds of the cucumber GY3 inbred line. The CUS1 gene, encoding the MADS-box type (agamus-like) protein, the expression pattern of which was described earlier, was used as a marker gene for optimisation of steps in the in situ RT-PCR inside the cells.

Molecular mechanism of TGF-beta1-induced apoptosis in HC11 mouse mammary epithelial cells (MEC).

The involvement of p53, Bax, cytochrome C and CPP-32 (caspase-3) in the molecular mechanism ofTGF-beta1-induced apoptosis in HC11 mouse mammary epithelial cells (MEC) was examined. Laser scanning cytometry (LSC) was applied for the quantitative analysis of expression and distribution of examined apoptosis-related proteins in the cytoplasmic (Cf) and nuclear (Nf) area. Maximal pixel of fluorescence (MP) parameter corresponding to aggregation of molecules in the cell was also measured.

Consumption of food products by the elderly living in different environments of the Warsaw region, Poland.

The aim of the study was to compare qualitatively the intake of food groups by the elderly living in urban and rural areas, with regard to the place of living and sex. Fifty-sex subjects in Warsaw and 142 in villages (born in 1919-24) participated in the study. A 3-day record method was used to collect nutritional data.

Production, purification and characterization of intracellular alanylaminopeptidase of Pseudomonas sp.

The soil bacterium Pseudomonas sp. was found to synthesize an aminopeptidase that prefers Ala-beta-naphtylamide as substrate. The enzyme was purified 660-fold by ammonium sulfate fractionation, preparative electrophoresis, ion exchange chromatography on Protein-Pak Q 8 HR and molecular sieving chromatography on Zorbax SE-250.

Medicine use and supplementation practice among participants of SENECA Study.

Objective: To assess six-year changes in medicine use and supplementation practice of elderly Europeans.

Design: Longitudinal assessment including two rounds of the SENECA STUDY: 1993 (first follow-up) and 1999 (final follow-up). Random sample of elderly born between 1913-1918, 284 men and 337 women in 1999.

Energy and nutrients intake of elderly people living in the Warsaw region, Poland.

The dietary intake of energy, macronutrients, minerals and vitamins was studied in a group of elderly living in urban, suburban, and rural areas of Warsaw region, Poland. The study was conducted in spring 1999 and 298 persons, born between 1919-1924, randomly selected with criteria of age and place of living, were included. Dietary data were based on the 3-day food records.

A simple, PC-dedicated, implanted digital PIM-radiotelemetric system. Part 2: The multichannel system.

The paper, which is a continuation of the previous one, describes a six-channel, PC-dedicated implanted telemetry system, including the schematic drawings, and explanation of all the differences between a single channel and multichannel system approach. The scheme of an additional multichannel analog output, that makes a pure analog recording possible, even without the PC connection, is also presented. Specific methods for both parallel (Centronics) and serial (RS232C) port interfacing are demonstrated and the controlling software principles are discussed.

Degradation of cellulose by nitrogen-fixing strain of Bacillus polymyxa.

During an intensive screening programme, several strains of cellulolytic bacteria were isolated. One nitrogenase-positive strain able to degrade filter paper, Avicel cellulose, carboxymethyl cellulose and cellobiose was selected for further study. On the basis of biochemical characteristics and Mol % of G+C content, the selected strain was identified as Bacillus polymyxa.

Pathogenesis of mousepox in H-2(d) mice: evidence for MHC class I-restricted CD8(+) and MHC class II-restricted CD4(+) CTL antiviral activity in the lymph nodes, spleen and skin, but not in the conjunctivae.

Genetically sensitive mice (i.e. H-2(d) haplotype) infected with a natural mouse pathogen named ectromelia virus (EV) can develop a mousepox.

Longevity and aging of mice from lines divergently selected for body weight for over 90 generations.

We have analysed the differences between longevity and aging process in two lines of mice selected divergently for body weight. As a result of long-term selection, the mice from two lines varied significantly in body weight during their life span. The differences regarded maturation rate, the length of the life time, reproduction performance, and fertility.

Enrichment of spreadable fats with polyunsaturated fatty acids omega-3 using fish oil.

Polyunsaturated fatty acids of the omega-3 series, especially very long chain--eicosapenta- and docosahexaenoic acid (EPA, DHA)--exert a strongly desirable influence on health. However, their intake with the western-style diet is usually too low which favours development of many diseases (CVD, cancers, allergies, etc.).

c-DNA vaccination against parasitic infections: advantages and disadvantages.

Recently developed technology for DNA vaccination appears to offer the good prospect for the development of a multivalent vaccines that will effectively activate both the humoral and cell mediated mechanisms of the immune system. Currently, DNA vaccination against such important parasitic diseases like malaria, leishmaniosis, toxoplasmosis, cryptosporidiosis, schistosomosis, fasciolosis offers several new opportunities. However, the outcome of vaccination depends very much on vaccine formulations, dose and route of vaccine delivery, and the species and even strain of the vaccinated host.

Subcellular redistribution of BAX during apoptosis induced by anticancer drugs.

BAX is the 192-amino acid, 21-kDa protein which is ubiquitously distributed in normal tissues and is regarded as a tumor suppressor sensitizing malignant cells to anticancer drugs. In spite of many studies, the molecular mechanism of BAX action is still obscure. In the present study subcellular BAX translocations in human colon adenocarcinoma COLO 205 cells exposed to various anticancer drugs [camptothecin (CPT), etoposide (ETO), staurosporine (STP), 2-chloro-2'-deoxyadenosine (2CdA) and nimesulide (NIM)] was examined.

The role of cholinergic and peptidergic pathways in the regulation of pancreatic exocrine function during postnatal development in pigs.

The aim of this study was to investigate the role of the parasympathetic (cholinergic and peptidergic) nervous system in the regulation of exocrine pancreas function in piglets during their early postnatal development. The cholinergic and peptidergic regulatory pathways of exocrine pancreatic function were tested by the specific muscarinic receptor blocker 4-diphenylacetoxy-N-methylpiperidine-methiodide (4-DAMP) and bombesin, respectively. At the age of 2 weeks, piglets were surgically fitted with a chronic pancreatic duct catheter, a duodenal re-entrant cannula and a jugular vein catheter.