Prognostic value of connexin43 expression in patients with clinically localized prostate cancer.

Connexins (Cxs) are a family of transmembrane proteins that build cell-to-cell channels in gap junctions. Gap junctions composed of Cxs have an essential role in intercellular communication, adhesion and cell differentiation. Several studies investigated the role of connexin43 (Cx43) in different carcinomas; however, none investigated its prognostic role in prostate cancer. Cx43 expression and relationship with established prognostic features were assessed in a cohort of 102 patients treated with radical prostatectomy for clinically localized prostate adenocarcinoma. Cx43 expression in prostate cancer was significantly associated with established features indicative of worse prognosis, such as follow-up time (P < 0.001) and preoperative PSA (P < 0.007). Patients with lower Cx43 expressions in tumours have shorter follow-up time, which indicated shorter disease-free survival and higher preoperative PSA values. Furthermore, tumours with positive surgical margins (P < 0.001) showed significantly lower Cx43 expression compared with tumours without this feature. In univariate (P < 0.001) and multivariate (P = 0.014) analyses, decreased Cx43 expression was found to be a significant predictor of biochemical recurrence free-survival. Study results show the association of decreased Cx43 expression with prostate cancer progression. Moreover, Cx43 could serve as an additional prognostic marker and used together with traditional prognostic markers might help in further stratifying the risk of disease progression in patients with prostate cancer.

[The effect of primary BHV-1 infection on the dynamics of T and B lymphocytes in the peripheral blood and levels of specific serum antibodies in calves treated with glucan]. / Efekt primárnej BHV-1 infekcie na dynamiku T a B lymfocytov periférnej krvi a hladinu specifických sérových protilátok u teliat osetrených glukánom.

In the present study we investigated the dynamics of circulating T and B lymphocytes and serum specific antibodies in calves experimentally infected with IBR virus (I group) and in calves administered glucan (seven days before infection) and the infected (GI group). The percentages of T and B lymphocytes in the peripheral blood were determined from analyses by rosette methods; the titer of serum anti-IBR antibodies was determined by virus-neutralizing test on cell cultures. The dynamics of the percentage of circulating T lymphocytes showed a similar decreasing trend in both groups, with significant values on days 3 to 5 after infection (AI). In comparison with the calves of I group, the outset of T cell reduction was found to be less pronounced (within the first two days after infection), with a statistically significant difference on day 2 AI (P < 0.05), Fig. 1. A decrease in the percentage of T lymphocytes was related to an increase in the percentage of circulating B lymphocytes, with maximum on days 3 and 4 AI (P < 0.05), Fig. 1. All the calves before experiment beginning were free of serum anti-BHV 1 antibodies. They started responding to the experimental infection with IBR virus by production of serum antibodies between week 1 and 2 AI. The dynamics of serum anti-IBR antibodies showed an identical course in both experimental groups, with the more pronounced outset (P > 0.05) of immunological response in the calves of GI group (Fig. 2).(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of bovine herpesvirus 1 with various types of DNA probes.

The method of dot-blot hybridization on nitrocellulose filters by various types of DNA probes (ds recombinant plasmids, ss recombinant M13 phages and a 42bp synthetic oligonucleotide) was used for BHV-1 detection. The highest sensitivity was achieved with the 32P-pUR1 probe (1.8 kb random EcoRI-HindIII fragment inserted into pUC9) which detected the BHV-1 genome in 5 x 10(3) infected MDBK cells. Using the pUR1 probe, no cross hybridization was observed with other herpesviruses: BHV-2, 3, 4, and Aujeszky's disease virus. The 32P-pUR1 probe detected BHV-1 in nasal swabs of calves as early as on day 1 after experimental infection. The maximum intensity of BHV-1 detection occurred on day 1-3. The 32P-pUR1 probe also detected BHV-1 in field samples of nasal swabs from cows and calves.

[Delayed hypersensitivity and the primary antibody reaction after administration of infectious bovine rhinotracheitis inactivated oil vaccine to calves premedicated with glucan]. / Uroven oneskorenej precitlivelosti a primárnej protilátkovej odpovede po aplikácii inaktivovanej olejovej IBR vakcíny u teliat premedikovaných glukánom.

The level of delayed skin hypersensitivity (DSH) to DNFB and of the primary immune reaction was evaluated in the calves immunized with an inactivated oil IBR vaccine (V group) and in the calves premedicated with glucan (seven days before vaccine administration) and subsequently immunized with the mentioned vaccine (GV group). The DSH test did not reveal an alteration of cellular immunological reactivity in the calves immunized with an inactivated oil IBR vaccine; after the vaccine administration there was no significant difference in the DSH level from the value before vaccine administration nor in comparison with the value of control calves (K group), Tab. I. But in the immunized calves which were glucan-premedicated (from Pleurotus ostreatus; 10 mg/kg l.w.) a significantly higher DSH level was determined not only in comparison with the initial value (P < 0.05) before glucan administration but also with the value of control calves (P < 0.05), Tab. I. The categorization of calves according to the DSH level also points to the immunomodulating effect of glucan in this sense (Fig. 1); while the value of skin test ranged from 3.6 to 6.5 mm in the highest percentage of the calves of V and K groups on the dates before and after administration of the mentioned preparations, a marked increase (on the date after administration) in the number of calves (from 22 to 67%) with the value of skin test higher than 6.5 mm was observed in the calves of GV group. The vaccine administration in itself (V group) did not induce the production of measurable amounts of serum IBR antibodies till day 14 after immunization.(ABSTRACT TRUNCATED AT 250 WORDS)

[Detection of bovine herpesvirus-1 using the dot-blot hybridization method]. / Detekcia bovinného herpesvírusu-1 metódou dot-blot hybridizácie.

With the development of molecular biology in the 1980s methods of microorganism detection start to innovate. One of the main advantages of the molecular-genetic methods, namely hybridization of nucleic acids and PCR methods, is the detection of genome of microorganism without the need for cellular cultivation. To detect BHV-1 (etiological agens IBR-IPV) the dot-blot hybridization method on nitrocellulose filters was used together with different types of DNA probes (two-fiber recombinant plasmids, one-fiber recombinant phages M 13 and 40 bp synthetic oligonucleotide). Genome DNA BHV-1 was isolated from samples (virions, infested cells, nasal smears and secretions by phenol extraction). The highest sensitivity of detection was achieved with 32P-pUR-1 probe (1.8kb random EcoRI-Hind III fragment ligated into plasmid pUC 9) which detected genome BHV-1 in 5 x 10(3) infested MDBK cells. This probe did not respond with herpetic viruses BHV-2, BHV-3, BHV-4 and the virus of Aujeszky's disease. The quality of pUR-1 probe was further tested for IBR diagnostics in animals experimentally infested with the virus BHV-1 (intranasal infection). BHV-1 could be detected in nasal smears and secretions in experimentally infested calves as early as on the first day following infection, while the agens amount reached its peak on the days 2-3 and on the days 6-7 the occurrence of virus fell markedly. When digoxigenin-pUR-1, i.e. non-radioactively marked probe, the virus presence was confirmed only on the days 2-3, in the time of the highest occurrence of infection agens. To detect the virus through the dot-blot hybridization nasal secretions were confirmed as better compared with nasal smears. The technology of virus isolation on cell cultures confirmed also the occurrence of agens as soon as on the first day from infection, with maximum on the days 2-5, but much more reliably it detected the virus on the days observed from the day 3 and their peak was obtained on the day 6 from infection. Experiments, comparing classical methods of IBR diagnostics (detection of specific antibodies, the method of isolation on cellular cultures) with the dot-blot hybridization using the samples obtained from farms with natural occurrence of IBM, are under progress.

[Preventive and metaphylactic use of Alga and Algalev in diarrheal and respiratory syndromes]. / Preventívne a metafylaktické pouzitie preparátov Alga a Algalev pri diarhoickom, resp. respiracnom syndróme.

Two gammaglobulin preparations have been developed: Alga (10% injection solution of serum bovine gamma-globulin and albumin) and Algalev (with addition of levamisole 10 mg/ml, patented under PV 4069/87); their administration to calves in field conditions was evaluated. The Alga preparation at a rate of 1 ml/kg liveweight (two administrations) had good preventive effects in the diarrhoeic syndrome of new-born hypogammaglobulinaemic calves. In comparison with the untreated calves, these parameters were determined: lower incidence of diarrhoea (41% against 75%; Tab. I), its later onset (by 24 hours), shorter duration (by 50 hours; P < 0.01), lower intensity and easier therapeutical handling (Tab. II). The Algalev preparation (two administrations at a rate of 1 ml/kg liveweight) was suitable for metaphylactic use (combined with antibiotic application) in the respiration syndrome of calves. In comparison with the untreated calves, these parameters were determined: lower incidence of this disorder (42% against 78%), lower intensity of clinical symptoms (intensive symptoms in 20% of the calves against 72%) and importantly higher daily weight gains (0.64 kg against 0.26 kg; Fig. 3). The concentrations of total serum Ig (CS-Ig), serum proteins (SB) and serum albumin (S-Alb) were similar in the compared groups of calves in the whole period of observation (Fig. 1). Certain differences were observed in the dynamics of anti-PI 3 titre of serum antibodies as shown by investigation of specific serum antibodies (Fig. 2); the level of their production was in agreement with morbidity incidence and clinical symptom intensity in both groups of calves.

Placental villous edema: a possible cause of antenatal hypoxia.

This study was undertaken to determine the relationship between the placental villous edema and the characteristic sequelae of antenatal hypoxia, i.e. the need for resuscitation at birth and low pH values in umbilical arterial blood. Placental villous edema was recognized by finding of open spaces within the cytoplasm of intervillous cells and in the interstitium of the villi. The percentage of edematous villi was significantly higher in the group of newborns requiring resuscitation. The severity of the edema had a positive correlation with the need for resuscitation at birth and with the arterial blood pH values in the umbilical cord. Placenta praevia and maternal toxicosis were associated with high percentage of edematous villi. It is suggested that edema fluid interposed a barrier to gas exchange between mother and fetus. The capillaries were blocked by compression leading to reduction in blood flow through the villi. These abnormalities, if widespread, may reduce gas exchange. It is suggested that hypoxia could partly be prevented by preventing the development of placental villous edema.

Alterations in monoamine oxidase activity of the mouse brain and liver after mixed neutron-gamma irradiation.

Monoamine oxidase (MAO) plays an important role in the metabolism of neuro-transmitter biogenic amines. Its activity was determined in mouse brain and liver after exposure to different kinds of ionizing radiation and after pretreatment with a radioprotective agent. After a lethal dose of mixed neutron-gamma irradiation the MAO activity decreased in the brain and increased in the liver. In contrast, after a lethal dose of 60Co-gamma irradiation enzyme activity was considerably increased in the brain while in the liver it increased like after mixed neutron-gamma irradiation. AET (S2-aminoethyl-isothiuronium-Br X HBr), when administered in a radio-protective dose, inhibited MAO activity in the brain, while it increased in the liver. Even more marked changes of enzyme activity were observed in both brain and liver after AET pretreatment and mixed neutron-gamma irradiation. On the basis of the results it is suggested that different kinds of ionizing radiation lead to different types of lipid peroxidation in the lipid environment surrounding MAO, an event leading to altered enzyme activity. AET itself inhibited MAO in the brain and increased the activity in the liver but did not prevent the alterations caused by ionizing radiation in enzyme activity.

Intrauterine growth of infants with chorioamnionitis.

The relationship between chorioamnionitis and fetal growth was examined by analysing the data of 299 infants with chorioamnionitis and the data of 296 infants with normal placentas. Six parameters were used for estimation of fetal growth: birth weight, length, head circumference, ponderal index, the ratio of length to head circumference and gestational age. Chorioamnionitis was identified, when at least 10 neutrophils per microscopic high-power field were present in the plate of placenta. There was no significant difference between the two groups at full term infants. The comparison could not be made under 37 weeks of gestation. It was concluded that chorioamnionitis probably did not impair particularly the fetal growth.

Acute 60Co-gamma irradiation of rats decreases the inhibitory effect of succinate on the lipid peroxidation of liver mitochondria.

Succinate inhibits NADPH-dependent lipid peroxidation of liver mitochondria. This effect of succinate decreased 12 h after whole-body 60Co-gamma irradiation, depending on the dose of irradiation.

The effect of superoxide dismutase on the radiosensitivity of bacterial spores at various oxygen concentrations.

Superoxide dismutases are considered to be essential protective agent against radiation injury. Experiments were carried out to investigate the effect of extracellularly added SOD on the radiosensitivity of Bacillus megaterium spores. 120 micrograms/ml SOD had no effect on the radiosensitivity of Bacillus megaterium spores at different oxygen concentrations. Relative enzyme activity obtained at various oxygen concentrations indicating the lack of oxygen effect in the radiation-induced inactivation of SOD.

Combination of misonidazole and lipoic acid on the radiation sensitivity of Bacillus megaterium spores.

The radiation sensitizing action of a combination of misonidazole and lipoic acid was investigated in Bacillus megaterium spores at various oxygen concentrations. Lipoic acid and misonidazole were applied in two concentrations (0.1 and 1.0 mmol/l) and four combinations were prepared from them. No uniform correlation was found, neither to the combination of the compounds nor to the gas conditions. The combination of two radiation sensitizing compounds with in all probability different action mechanisms does not unequivocally enhance the radiation sensitizing effect under anoxic and hypoxic conditions.

Effect of acute 60Co-gamma-irradiation on the in vivo lipid peroxidation in experimental animals.

The effect of sublethal (6.0 Gy) and LD50/30 (9.0 Gy) doses of 60Co-gamma-irradiation on malondialdehyde (MDA) level was studied in rats. The findings suggest that in the organs investigated (brain, liver, spleen, kidney, testicle, stomach, small intestines) acute 60Co-gamma-irradiation increased the formation of MDA, the main product of lipid peroxidation, in a time-related manner to an extent characteristic of the organ investigated. Differences in the degree and temporal development of the changes allow some conclusions as to the radiosensitivity of individual organs. On this basis it can be assumed that the increase in MDA level caused by irradiation considerably contributes to the development of certain symptoms of radiation sickness.

Study of "prostacyclin-like" activity in the rat blood vessel: changes in prostacyclin production related to body weight.

The prostacyclin-producing capacity of various blood vessels was studied using platelet aggregation bioassay in rats of different ages. Results showed that --the PGI2 producing capacity of arteries was five to tenfold of the respective veins; --in the arteries the PGI2 production calculated for unit vascular mass declined from the centre to the periphery, despite the relative increase of endothelial surface area. This points to the importance of other vascular elements in the control of PGI2 production; --the PGI2 forming activity of some blood vessels increased with the age up to a limit (300-400 g body weight), decreasing thereafter. Based on these results we conclude that major part of PGI2 in systemic blood originates from the large arteries, whereas the low PGI2 production of the veins might play a role, among others, in the more frequent occurrence of venous thrombosis.