Redistribution of Cs 137 introduced into montmorillonite in association with organic matter coming from biomass composting.

The adsorption and later bioavailability of 137Cs from the system humic acid (HA)/humic acid like compounds (HALC) and montmorillonite was investigated. The setup of the experiments should approach as much as possible natural conditions when 137Cs is introduced into soil with HALC from decomposed biomass. The significant differences were found in the trials containing various HA/HALC and also pure montmorillonite. The 137Cs was more available when it reached soil in association with HALC originated from compost than when it was adsorbed on stable humic acids. Moreover, the long term interaction of 137Cs with HALC led to decrease of 137Cs adsorbed on montmorillonite and increase of its bioavailable fraction. UV-Vis spectrometry and infrared spectroscopy showed the clear difference between HA, fresh HALC and old HALC which could partially explain the different results.

Variation of ²¹°Po daily urinary excretion for male subjects at environmental level.

(210)Po was determined in 24-h urine of seven healthy males from Prague, Czech Republic, for ten consecutive days. The results show that for each volunteer, the urinary excretion of (210)Po changed only little from day to day in the studied time period. For two volunteers, the difference in the daily excreted (210)Po activity for two consecutive days was not significant, given the 95% confidence interval (two sigma) of the activity measurements. The same is valid for the excretion data of the other volunteers, except for some days where the differences were slightly higher. The range of daily urinary excretion of (210)Po of each volunteer in the studied time period was quite narrow. Among the volunteers, the maximum daily urinary excretion value of (210)Po was at most about a factor of 2.5 higher than the lowest excretion value. An attempt to explain the observed small inter-individual variability of (210)Po excretion in daily urine is made.

[Interaction between p53 and MDM2 in human lung cancer cells]. / Interakcia medzi p53 a MDM2 v nádorových bunkách karcinómu plúc.

BACKGROUND: The oncoprotein p53 protein induces cell growth arrest (apoptosis) in response to endo  or exogenous stimuli. Mutation of TP53 (gene encoding the p53 protein) is common in human malignancies and alters the conformation of p53. The result is a more stable protein which accumulates in nuclei of tumor cells with loss of function. Mutant p53 is stabilized, and it is possible to detect this form very clearly by immunohistochemistry (IHC). Expression of the MDM2 protein is used as a potential marker of p53 function. P53 levels in normal cells are highly determined by the MDM2 protein (murine double minute 2) -  mediated degradation of p53. MDM2 overexpression represents at least one mechanism by which p53 function can be abrogated during tumorigenesis. MATERIAL AND METHODS: Lung carcinoma samples were obtained from patients, who underwent radical resection (lobectomy or pulmonectomy and lymphadectomy). Pathological dia-gnosis was based on the WHO criteria. In our study, we investigated the expression of p53 and MDM2 protein that might improve IHC as a marker for p53 status. Proteins were IHC detected in 136 samples of primary lung carcinoma. Immunostaining results of p53 positive samples were compared to IHC expression of MDM2 positive and MDM2 negative samples. RESULTS: Strong brown nuclear staining was visible in p53 and MDM2 positive cells. The most p53 positive cases were samples of squamocellular carcinoma (55%), then samples of large cell carcinoma (53%) and 26% adenocarcinoma samples showed the p53 immunoreactivity. No one sample of different types was p53 positive. When we compared the p53 expression and grade of tumor, we found that the p53 expression increased with the grade of tumor. For statistical evaluation, the chi square test was used. The relationship between p53 expression and type of tumor, also the p53 expression and grade of tumor was statistically significant (p = 0.000425; p = 0.00157). Regarding p53 and MDM2 expression, only nine samples (7%) were simultaneously p53 and MDM2 positive. In 46 (34%) cases, elevation of p53 was combined with MDM2 negative expression. Other tumor samples were either negative for both proteins (71/ 52%), or p53 negative and MDM2 positive in 10 (7%) tumor samples. CONCLUSION: Absence of p53 staining in most studies indicates absence of p53 mutation, and on the contrary, positive expression of p53 is a sign of p53 mutations with loss of function. In our study, 34% of cases with extensively high level of p53 without increased level of MDM2 were identified. We suppose that these are tumors with inactivating mutations that stabilize p53. On the other hand, tumors with high level of stabilized wildtype p53 protein and simultaneously with increased MDM2 staining (9 samples/7%) represent group with functional p53. In this group of patients, we could expect better prognosis with regard to function of p53 protein.

[Importance of expression of DNA repair proteins in non-small-cell lung cancer]. / Význam stanovovania expresie DNA reparacných mechanizmov u nemalobunkového karcinómu plúc.

BACKGROUND: Proteins XRCC1 and ERCC1 are involved in DNA repair. XRCC1 plays a role in DNA base excision repair and ERCC1 in nucleotide excision repair pathway. Higher expression profile of both proteins in cancer cells may contribute to development of drug resistance. ERCC1 is involved in removal of platinum adducts and might be a potential predictive and prognostic marker in NSCLC (non-small-cell lung cancer) treated with a cisplatin-based regimen. The purpose of study was determination of XRCC1 and ERCC1 levels and their correlation with basic clini-copathological parameters in NSCLC. PATIENTS AND METHODS: In this study, 107 tumor samples diagnosed as NSCLC were immunohistochemically examined for expression of XRCC1 and ERCC1 proteins. Our results were compared to basic clinicopathological parameters: type of tumor, tumor grade and stage of disease. For statistical analysis, the chi-square test was used. RESULTS: In squamous cell carcinoma and large cell carcinoma samples, the XRCC1 protein level was twofold higher (60% of positive samples) than in adenocarcinoma samples (35.5% of positive samples). We have found statistical correlation between XRCC1 protein expression and type of tumor (p = 0.0306). On the other hand, the statistical importance between the protein level versus grade and stage was not found. In the case of the ERCC1 protein, we observed the highest protein level in adenocarcinoma (64.5%) and squamous cell carcinoma (62.5%) samples. Next, we determined a significant difference in content of XRCC1 versus ERCC1 (35.5% vs 64.5%) in adenocarcinoma samples. Statistical chi-square test did not reveal any correlation between ERCC1 status and clinicopathological parameters. CONCLUSION: According to our results, XRCC1 represents an important mechanism of DNA repair in squamous cell and large cell carcinomas. Besides that, expression of XRCC1 was in correlation with type of tumor. In patients with adenocarcinoma and squamous cell carcinoma, we could assume increased resistance to platinum-based therapy because of high expectation of ERCC1 protein expression. However, its levels did not correlate with monitored clinicopathological parameters. The ERCC1 protein will be possibly an independent prognostic factor in NSCLC. To prove a true survival benefit of patients with expression of ERCC1, prospective validation of ERCC1 before clinical implication is needed in the future.

Proceedings from the Ice Hockey Summit on concussion: a call to action.

The objective of this proceedings is to integrate the concussion in sport literature and sport science research on safety in ice hockey to develop an action plan to reduce the risk, incidence, severity, and consequences of concussion in ice hockey. A rationale paper outlining a collaborative action plan to address concussions in hockey was posted for review two months prior to the Ice Hockey Summit: Action on Concussion. Focused presentations devoted specifically to concussion in ice hockey were presented during the Summit and breakout sessions were used to develop strategies to reduce concussion in the sport. This proceedings and a detailed scientific review (a matrix of solutions) were written to disseminate the evidence-based information and resulting concussion reduction strategies. The manuscripts were reviewed by the authors, advisors and contributors to ensure that the opinions and recommendations reflect the current level of knowledge on concussion in hockey. Six components of a potential solution were articulated in the Rationale paper and became the topics for breakout groups that followed the professional, scientific lectures. Topics that formed the core of the action plan were: metrics and databases; recognizing, managing and return to play; hockey equipment and ice arenas; prevention and education; rules and regulations; and expedient communication of the outcomes. The attendees in breakout sessions identified action items for each section. The most highly ranked action items were brought to a vote in the open assembly, using an Audience Response System (ARS). The strategic planning process was conducted to assess: Where are we at?; Where must we get to?; and What strategies are necessary to make progress on the prioritized action items? Three prioritized action items for each component of the solution and the percentage of the votes received are listed in the body of this proceedings.

Histochemical detection of monoamine oxidases in rat female genital organs during preimplantation period of pregnancy.

OBJECTIVE: Localization of monoamine oxidases (MAO) in rat female gonads during preimplantation period of pregnancy was determined. MATERIAL AND METHODS: Pregnant females were killed on their first, third, and fifth days of pregnancy and animals were transcardially perfused with PBS and fixative solutions. Ovaries, oviducts and uteri were immediately removed and they served for the determination of MAO localization employing the method of enzymatic histochemistry. RESULTS: MAO-A activity in ovary was visible in corpora lutea and in interstitial gland cells while MAO-B was detected predominantly in blood vessels. Both MAO enzymes were seen in the smooth muscle fibers of the ovarian hilum. The presence of MAO enzymes was however not detected in follicles at any stage of their development. In oviduct and uterus, both MAO enzymes were visible in similar places, namely in smooth muscle fibers, mast cells and blood vessels, with no MAO presence seen in the epithelium. CONCLUSIONS: Potential physiological importance of MAO localization in different cells of female reproductive organs during early period of pregnancy is proposed (Fig. 6, Ref. 29).

Relationship between serotonin and norepinephrine levels and the preimplantation embryo development in rat.

OBJECTIVE: We have evaluated the impact of chronic administration of clorgyline, a potent monoamine oxidase A inhibitor and a former antidepressant, on the preimplantation embryo development in Wistar rats. MATERIAL AND METHODS: Females were injected intraperitoneally daily for 30 days with saline (control animals, C), or with a low dose of clorgyline (0.1 mg/kg/day, LDC) or with a high dose of clorgyline (1 mg/kg/day, HDC). Embryos were isolated on day 5 of pregnancy and their urine was collected. RESULTS: The number of embryos per female did not differ between experimental groups and control, but we have recorded a decreased number of embryos in HDC group compared to LDC (p < 0.05). We have found that LDC significantly reduced the presence of healthy embryos and increased the presence of the degenerated embryos (p < 0.001). The administration of the LDC resulted in the lowest cell number in blastocysts (p < 0.01). Concerning monoamines in urine, we have observed significantly increased serotonin levels in HDC group compared to control (p < 0.05) and LDC animals (p < 0.01). Norepinephrine levels in both experimental groups were significantly elevated compared to controls, too (p < 0.001 LDC vs C; p < 0.01 HDC vs C). CONCLUSIONS: We assume that the impaired embryo development recorded in the LDC group after clorgyline administration was a consequence of the higher levels of the norepinephrine. We speculate that lesser negative effect of HDC compared to LDC on the preimplantation embryo development could be the consequence of the lesser norepinephrine levels and/or elevated serotonin levels (Tab. 2, Fig. 2, Ref. 37). Full Text in free PDF www.bmj.sk.

Immunohistochemical evaluation of Pi class glutathione S-transferase expression in invasive breast carcinoma.

OBJECTIVES: The aim of our work was to determine the expression of Pi class glutathione S-transferase (GSTP1) in 43 samples of invasive breast carcinoma and compare results versus normal breast cells. BACKGROUND: Breast cancer is the commonest cancer in women. Despite advances in early detection and more efficacious adjuvant chemotherapy, a part of patients with early-stage have reccurent disease. In these cases the development of resistance to therapy is observed. The glutathione S-transferases (GSTs) are potentially involved in tumour chemoresistance. METHODS: Enzyme immunohistochemical method was chosen for the detection of GSTP1 and its expression was compared in breast cancer cells versus normal breast cells. RESULTS: We have found that majority (63%) of breast carcinomas shows GSTP1 positivity (nuclear and cytoplasmic immunoreactivity). It is expected that GSTP1 positive tumours would show a poorer prognosis than GSTP1 negative ones. CONCLUSION: Immunohistochemistry is a useful method for investigating the expression and cellular localization of GSTP1 within tumours. It may be applied to a routine clinical test and it can serve as a marker for resistance to chemotherapy (Tab. 1, Fig. 4, Ref. 20). Full Text in free PDF www.bmj.sk.

Antagonistic effect of low deprenyl dose on the preimplantation embryo development in rat.

To investigate the role of potent MAO-B inhibitor deprenyl in fertilized females, we have evaluated the effect of chronic treatment with deprenyl at a low dosage on preimplantation embryo development in Wistar rats. We have found that the number of isolated embryos per rat did not differ between experimental and control groups. But morphological analysis of embryos isolated from deprenyl-treated animals had revealed improved rates in the distribution pattern compared with controls. On the other hand, harmful impact of deprenyl administration on the mean cell number in blastocysts and on their cell proliferation has been recorded. To our knowledge this is the first paper describing antagonistic effect of deprenyl administration on the preimplantation embryo development in mammals. Potential mechanisms mediating biphasic deprenyl-induced impact on embryonic development are proposed (Tab. 3, Ref. 17). Full Text (Free, PDF) www.bmj.sk.

Effect of exercise on the caloric intake of breast cancer patients undergoing treatment.

The purpose of this study was to examine the effects of an exercise intervention on the total caloric intake (TCI) of breast cancer patients undergoing treatment. A secondary purpose was to determine whether or not a relationship existed between changes in TCI, body fat composition (%BF), and fatigue during the study, which lasted 6 months. Twenty females recently diagnosed with breast cancer, scheduled to undergo chemotherapy or radiation, were assigned randomly to an experimental (N = 10) or control group (N = 10). Outcome measures included TCI (3-day food diary), %BF (skinfolds), and fatigue (revised Piper Fatigue Scale). Each exercise session was conducted as follows: initial cardiovascular activity (6-12 min), followed by stretching (5-10 min), resistance training (15-30 min), and a cool-down (approximately 8 min). Significant changes in TCI were observed among groups (F1,18 = 8.582; P = 0.009), at treatments 2 and 3, and at the end of the study [experimental (1973 +/- 419), control (1488 +/- 418); experimental (1946 +/- 437), control (1436 +/- 429); experimental (2315 +/- 455), control (1474 +/- 294), respectively]. A significant negative correlation was found (Spearman rho(18) = -0.759; P < 0.001) between TCI and %BF and between TCI and fatigue levels (Spearman rho(18) = -0.541; P = 0.014) at the end of the study. In conclusion, the results of this study suggest that an exercise intervention administered to breast cancer patients undergoing medical treatment may assist in the mitigation of some treatment side effects, including decreased TCI, increased fatigue, and negative changes in body composition.

Effect of exercise on the caloric intake of breast cancer patients undergoing treatment

The purpose of this study was to examine the effects of an exercise intervention on the total caloric intake (TCI) of breast cancer patients undergoing treatment. A secondary purpose was to determine whether or not a relationship existed between changes in TCI, body fat composition ( percentBF), and fatigue during the study, which lasted 6 months. Twenty females recently diagnosed with breast cancer, scheduled to undergo chemotherapy or radiation, were assigned randomly to an experimental (N = 10) or control group (N = 10). Outcome measures included TCI (3-day food diary), percentBF (skinfolds), and fatigue (revised Piper Fatigue Scale). Each exercise session was conducted as follows: initial cardiovascular activity (6-12 min), followed by stretching (5-10 min), resistance training (15-30 min), and a cool-down (approximately 8 min). Significant changes in TCI were observed among groups (F1,18 = 8.582; P = 0.009), at treatments 2 and 3, and at the end of the study [experimental (1973 ± 419), control (1488 ± 418); experimental (1946 ± 437), control (1436 ± 429); experimental (2315 ± 455), control (1474 ± 294), respectively]. A significant negative correlation was found (Spearman rho(18) = -0.759; P < 0.001) between TCI and percentBF and between TCI and fatigue levels (Spearman rho(18) = -0.541; P = 0.014) at the end of the study. In conclusion, the results of this study suggest that an exercise intervention administered to breast cancer patients undergoing medical treatment may assist in the mitigation of some treatment side effects, including decreased TCI, increased fatigue, and negative changes in body composition.

Study of high deprenyl dose on the preimplantation embryo development and lymphocyte DNA in rat.

To investigate the role of potent MAO-B inhibitor deprenyl in fertilized females, we have evaluated the effect of chronic treatment with deprenyl at a high dosage on preimplantation embryo development and DNA damage in blood lymphocytes in Wistar rats. We have found that the number of embryos isolated from both uterus and oviduct per rat was significantly lower in the experimental group. Almost 14% of embryos in experimental animals were flushed from oviducts compared to 1.95% of those in the control rats. Morphological analysis of embryos isolated from deprenyl-treated animals had revealed impaired rates in the distribution pattern compared with controls. But deprenyl administration had no significant effect on the mean number of cells in morulae or even blastocysts. On the other hand, analysis of cell number distribution in blastocysts using the chi-square test indicated a significantly decreased cell proliferation in the experimental group. Despite the harmful impact of deprenyl on rat preimplantation embryo development, deprenyl administration significantly decreased the DNA damage in blood lymphocytes as was scored employing Comet Assay. Our description of the adverse effects of deprenyl administration on rat preimplantation embryo development compared to the protective effects on the lymphocyte DNA is very important because deprenyl is still widely used in human medicine as a treatment. Potential mechanisms mediating deprenyl-induced impaired preimplantation embryo development are proposed.

Multidrug resistance proteins in renal cell carcinoma.

A large number of renal cancer patients show poor or partial response to chemotherapy and the precise mechanism has not been understood yet. MDR is the principal mechanism by which many cancers develop resistance to chemotherapeutic drugs and is associated with the elevated expression of MDR proteins. These are divided into two groups: ABC transporters and non-ABC transporters. The aim of our study was to determine the expression of MDR1/Pgp, MRP1 and LRP in 47 samples of renal cell carcinomas using immunohistochemical assay. Our results were analysed in relation to nuclear grade and other clinical and pathological parameters to see the possible correlation between the expression of MDR proteins and factors mentioned above. The majority of renal carcinoma specimens showed positivity for MDR proteins. In this regard, 21 % of samples revealed positive results for MDR1, 62 % for MRP1 and 76.6 % for LRP protein. Furthermore, our study displayed significant differences between MDR1, LRP and nuclear grade. On the other hand, no association was found between MRP1 and nuclear grade, as well as between the expression of three MDR proteins and other clinically relevant parameters.

Expression of serotonin receptors in mouse oocytes and preimplantation embryos.

Serotonin receptors have been found in several reproductive organs as well as in the central nervous system. Serotonin-binding sites have been demonstrated in duck ovarian follicles and the testis, hamster ovaries, human granulosa cells and mouse placenta. Local production of serotonin by the rat ovary, oviduct, uterus and testis has also been reported. We analyzed the expression of three types of serotonin receptors: 5-HT1B, 5-HT2C and 5-HT1D by reverse transcription-polymerase chain reaction in mouse unfertilized oocytes and preimplantation embryos from zygotes to the blastocyst stage in vivo. Transcripts for 5-HT1B and 5-HT2C serotonin receptors were detected neither in unfertilized oocytes nor at any stages of in vivo developing preimplantation embryos. Serotonin 5-HT1D receptor mRNA was present in unfertilized oocytes, zygotes, 2-cell embryos, compacted morulae and in vivo produced expanded blatocysts. The expression of the mRNA 5-HT1D serotonin receptor was also detected in blastocysts cultured in vitro. When added to the culture medium, specific serotonin 5-HT1D agonist sumatriptan (1 microM) significantly inhibited the development of mouse embryos cultured in vitro. Demonstration of the expression of 5-HT1D serotonin receptor in mouse oocytes and preimplantation embryos supports the idea of a functional serotonin (5-HT1D) receptor in early mammalian development.

Accumulation of the proteolytic marker peptide ubiquitin in the trophoblast of mammalian blastocysts.

Ubiquitination is a universal protein degradation pathway in which the molecules of 8.5-kDa proteolytic peptide ubiquitin are covalently attached to the epsilon-amino group of the substrate's lysine residues. Little is known about the importance of this highly conserved mechanism for protein recycling in mammalian gametogenesis and fertilization. The data obtained by the students and faculty of the international training course Window to the Zygote 2000 demonstrate the accumulation of ubiquitin-cross-reactive structures in the trophoblast, but not in the inner cell mass of the expanding bovine and mouse blastocysts. This observation suggests that a major burst of ubiquitin-dependent proteolysis occurs in the trophoblast of mammalian peri-implantation embryos. This event may be important for the success of blastocyst hatching, differentiation of embryonic stem cells into soma and germ line, and/or implantation in both naturally conceived and reconstructed mammalian embryos.

The influence of insulin on the in vitro development of mouse and bovine embryos.

To further investigate the role of insulin during preimplantation embryo development, we compared the effects of insulin on the development of mouse and bovine preimplantation embryos and on cell proliferation during culture in vitro in simplex media. The influence of insulin on the development of mouse zygotes was determined during cultivation in mSOF medium, alone or supplemented with glucose. Similarly, the effects of insulin on the bovine preimplantation embryo development were studied in mSOF medium. The addition of insulin into mSOF medium enhanced significantly the number of cells per mouse blastocyst. Moreover, when mSOF medium was supplemented with insulin and 0.2 mmol x l(-1) glucose, the percentage of hatched blastocysts and the mean cell number of mouse blastocysts were significantly higher. Insulin had no significant effect on the development of bovine embryos, produced by in vitro fertilization of in vitro matured oocytes. Neither the rates of developing embryos nor the mean number of cells in blastocysts were different in comparison with control embryos. Our results suggest that the in vitro development of mouse embryos could be enhanced by the addition of insulin to the culture medium and is further improved by the addition of glucose. In contrast to this our results indicate that insulin has no detectable beneficial effect on the preimplantation development of bovine embryos in mSOF medium.

Preimplantation embryo development in ICR mice after streptozotocin treatment.

To investigate the significance of impaired insulin secretion on preimplantation embryo development, outbred ICR female mice received a single injection of streptozotocin 130 mg (low) and 160 mg (subdiabetic) kg(-1), 14-17 days before fertilization. Preimplantation embryos were collected on day 3 of pregnancy, four to eight-cell embryos were cultured in vitro 48 h (day 5) and their cell number was estimated. After spontaneous ovulation, the significantly different distribution pattern in comparison with the controls was detected only in preimplantation embryos isolated from subdiabetic (160 mg x kg(-1) streptozotocin) mice. Furthermore, the incidence of degenerated embryos was significantly increased after 48 h in vitro cultivation. The analysis of cell number distribution in embryos after cultivation in vitro indicated a significant delay in cell proliferation in both experimental groups (130 and 160 mg x kg(-1) streptozotocin) in comparison with control mice. After superovulation, the only significant difference was found in the distribution pattern of embryos isolated on day 3 of pregnancy from subdiabetic (160 mg x kg(-1) streptozotocin) mice. No significant differences were found after embryo cultivation in vitro. It could be concluded that, in outbred ICR mice, lower streptozotocin treatment (130 mg x kg(-1)) influenced only cell distribution of in vitro cultured embryos after spontaneous ovulation. In ICR mice, marked changes in preimplantation embryo development were detected only after subdiabetic (160 mg x kg(-1)) streptozotocin treatment. During in vitro cultivation delayed effects of impaired insulin secretion resulted in an increase of embryo degeneration at the time after the third mitotic cleavage. Our results indicate that the effects of impaired maternal insulin secretion on preimplantation embryo development in mice are marked and consistent after spontaneous ovulation. Superovulation apparently disguises subtle changes in preimplantation embryo development after low and subdiabetic streptozotocin treatment.

Effects of impaired maternal insulin secretion on preimplantation embryo development in ICR mice.

To investigate the significance of impaired insulin secretion on preimplantation embryo development, outbred ICR female mice received an injection of a single dose of streptozotocin 200 mg.kg-1 14-17 days before fertilization. Oocytes were collected 24-26 h after hCG injection. Morphological evaluation revealed a lower percentage of oocytes with second polar bodies from streptozotocin-treated females in comparison with controls. Furthermore, in this group the incidence of degenerated embryos significantly increased after 120 h in vitro cultivation. Insulin (5 U per 100 g b.w.) administered twice daily to streptozotocin-treated mice significantly improved the Embryonic development. Morphological analysis of oocyte maturation in streptozotocin-treated mice showed no significant differences in comparison with control mice. It could be concluded that marked changes in preimplantation embryo development were detected in outbred ICR mice after streptozotocin administration and this process was partly reversible by insulin treatment. Furthermore, it was shown that the process of fertilization was negatively influenced and that during in vitro cultivation the delayed effects of impaired insulin secretion resulted in an increase of embryo degeneration at the time following the third mitotic cleavage.

Legal aspects of forensic psychiatry in South Africa.

The current political changes in South Africa have focused attention on matters such as the availability of medical and psychiatric services for all. These services have been stretched to breaking point due to waves of crimes of violence which seem to go in tandem with the process of democratization. These questions are discussed and some possible inadequacies of the forensic sciences in the criminal justice system are considered.