Human brain thioltransferase: constitutive expression and localization by fluorescence in situ hybridization

Thioltransferase (glutaredoxin) is a member of the family of thiol-disulfide oxido-reductases that maintain the sulfhydryl homeostasis in cells by catalyzing thiol-disulfide interchange reactions. One of the major consequences of oxidative stress in brain is formation of protein-glutathione mixed disulfide (through oxidation of protein thiols) which can be reversed by thioltransferase during recovery of brain from oxidative stress. Here we have visualized the location of thioltransferase in brain regions from seven human tissues obtained at autopsy. Constitutively expressed thioltransferase activity was detectable in all human brains examined although inter-individual variations were seen. The enzyme activity was significantly higher in hippocampus and cerebellum as compared to other regions. Constitutive expression of thioltransferase mRNA was detectable by Northern blot analysis. Localization of thioltransferase mRNA by fluorescence in situ hybridization revealed its presence predominantly in neurons in the cerebral cortex, Purkinje and granule cell layers of the cerebellum, granule cell layer of the dentate gyrus and in the pyramidal neurons of CA1, CA2 and CA3 subfields of hippocampus. These discrete neuronal concentrations of thioltransferase would be consistent with an essential role in modulating recovery of protein thiols from mixed disulfides formed during oxidative stress.     

Reduction of atherosclerosis in cholesterol-fed rabbits and decrease of expressions of intracellular adhesion molecule-1 and vascular endothelial growth factor in foam cells by a water-soluble fraction of Polygonum multiflorum

Polygonum multiflorum stilbeneglycoside （PMS） is a water-soluble fraction of Polygonum multiflorum Thunb. , one of the most famous tonic traditional Chinese medicines, that has protective effects on the cardiovascular system. The purpose of the present study is to elucidate the effects of PMS on macrophage-derived foam cell functions and the reduction of severity of atherosclerosis in hypercholesterolemic New Zealand White （NZW） rabbits. NZW rabbits were fed for 12 weeks with a normal diet, a high cholesterol diet, or a high cholesterol diet associated with irrigation with different doses of PMS （25, 50, or 100 mg/kg）. Treatment of NZW rabbits fed with high cholesterol diet with 100 mg/kg PMS attenuated the increase in plasma cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, and plasma triglyceride. Treatment with 50 and 100 mg/kg PMS caused 43% and 60% decrease in atherosclerotic lesioned area ratio to total surface area, respectively. In U937 foam cells, PMS could decrease the high expression of intercellular adhesion molecule （ICAM）-1 protein and the vascular endothelial growth factor （VEGF） protein levels in the medium induced by oxidized lipoprotein when analyzed by flow cytometry. The results proved that PMS is a powerful agent against atherosclerosis and that PMS action could possibly be through the inhibition of the expression of ICAM-1 and VEGF in foam cells.     

Three-dimensional video-assisted thoracoscopic pericardiectomy

Video-assisted thoracic surgery has proven to be safe and effective for the diagnosis and management of pericardial disease. Three-dimensional (3-D) video imaging technology has been developed to allow the laparoscopic surgeon more precision and efficiency in advanced laparoscopic cases. This case report describes the marriage of 3-D video imaging and thoracoscopy that allowed performance of a technically difficult pericardiectomy without incident. Our aim is to describe the use of state-of-the-art 3-D video imaging to allow success in difficult cases.     

Immunotoxic effects of short-term atrazine exposure in young male C57BL/6 mice.

The herbicide atrazine (ATR) is a very widely used pesticide; yet the immunotoxicological potential of ATR has not been studied extensively. Our objective was to examine the effect of ATR on selected immune parameters in juvenile mice. ATR (up to 250 mg/kg) was administered by oral gavage for 14 days to one-month-old male C57BL/6 mice. One day, one week, and seven weeks after the last ATR dose, mice were sacrificed, and blood, spleens, and thymuses were collected and processed for cell counting and flow cytometry. Thymus and spleen weights were decreased by ATR, with the thymus being more sensitive than the spleen; this effect was still present at seven days, but not at seven weeks after the last ATR dose. Similarly, organ cellularity was persistently decreased in the thymus and in the spleen, with the splenic, but not thymic cellularity still being depressed at seven weeks post ATR. Peripheral blood leukocyte counts were not affected by ATR. There were also alterations in the cell phenotypes in that ATR exposure decreased all phenotypes in the thymus, with the number of CD4(+)/CD8(+) being affected the least. At the higher doses, the decreases in the thymic T-cell populations were still present one week after the last ATR dose. In the spleen, the CD8(+) were increased and MHC-II(+) and CD19(+) cells were decreased one day after the last ATR dose. Also, ATR treatment decreased the number of splenic na?ve T helper and T cytotoxic cells, whereas it increased the percentage of highly activated cytotoxic/memory T cells. Interestingly, the proportion of mature splenic dendritic cells (DC; CD11c(high)), was also decreased and it persisted for at least one week, suggesting that ATR inhibited DC maturation. In the circulation, ATR exposure decreased CD4(+) lymphocytes at one day, whereas at seven days after the last ATR dose, in addition to the decrease in CD4(+) lymphocytes, the MHC-II(+) cells were also decreased at the 250 mg/kg dose. Thus, ATR exposure appears to be detrimental to the immune system of juvenile mice by decreasing cellularity and affecting lymphocyte distribution, with certain effects persisting long after exposure has been terminated.     

Expression of integrin adhesion molecules in normal ovary and epithelial ovarian tumors

The metastatic potential of a solid tumor is dependent upon its ability to interact with the extracellular matrix. The integrin superfamily is a group of proteins that are fundamental in such interactions and play a major role in cell-cell and cell-matrix adhesion. Localization of the integrin proteins was performed in normal ovary, primary epithelial ovarian tumors and metastatic tumor cells in ascitic samples. Expression of α₁, α₃, α₆ and β₄ was observed on normal ovarian epithelium with variable expression of α₅. Loss of α₁ expression by malignant cells in the primary tumors was noted. β₄, a component of the laminin receptor which was strongly expressed by both normal ovary and solid tumor, was absent from the ascitic tumor cells in the majority of cases. There was an associated loss of α₆ expression, indicating a deficiency of hemidesmosomes in the ascitic tumor cells. This alteration of integrin expression by metastatic malignant epithelial ovarian tumor cells may therefore represent one important mechanism by which metastatic disease occurs.     

Enhanced thromboxane B2 release from challenged guinea pig lung after oxygen exposure

The effects of exposure to 100% oxygen (O2) on the release of thromboxane B2 (TXB2) and prostaglandins (PG) from guinea pig lungs during anaphylaxis were studied. Lungs from sensitized guinea pigs were isolated, perfused, and challenged. Serologic examination of the perfusate showed that larger amounts of TXB2, 6-keto-PGF1 alpha, PGE2, and PGF2 alpha were released from the lungs of animals previously exposed to 100% O2 for 72 h. Conversely, these lungs released smaller amounts of 13, 14-dihydro-15-keto-PGF2 alpha. Histamine release, measured spectrofluorometrically, was unchanged. 15-hydroxyprostaglandin dehydrogenase (PGDH) was inhibited by 83% after O2 exposure, as measured by an in vitro enzyme assay. These results suggested that the reported enhancement of systemic anaphylaxis in the guinea pig in vivo after exposure to oxidant gases may be due in part to inhibition of pulmonary PGDH resulting in increased half-lives of primary PG and TX released from the lung.     

THE EFFECT OF MULTIPLE EMBOLI OF THE CAPILLARIES AND ARTERIOLES OF ONE LUNG

1. Injection of a suspension of potato starch cells into the left branch of the pulmonary artery, in quantity sufficient ordinarily to give rise to markedly accelerated respirations, resulted in no change in respiratory rate. 2. A method for injecting substances into the pulmonary artery or its branches without interfering with the blood flow to the lungs has been described. 3. Injection of similar material into one lung when the other is excluded from the circulation either by ligation or by temporary clamping does give rise to rapid and shallow breathing (from a rate of 10 to 15 per minute to one of 60 or over) identical in character to that brought about by introducing emboli into both lungs. 4. A method for clamping and releasing the pulmonary artery or its branches in a dog breathing normally with closed thorax has been devised. This is described in detail in another paper. 5. After rapid breathing has been initiated by the effect of emboli lodged in the arterioles and capillaries of the right lung, reestablishing the circulation in the other lung by releasing the clamp on its artery may or may not restore the respiratory rate to its original, normal level. 6. This discrepancy in results has not been correlated with any difference in oxygen saturation of the arterial blood, or in carbon dioxide tension or pH of its plasma. 7. It is, however, believed to be related to the gross and microscopic anatomy of the lung of which the artery has been temporarily clamped. Photomicrographs are published, showing in one dog (No. 3), in which the respiratory rate returned to normal, a normal histological picture of the left lung, and in another dog (No. 4), in which the rate remained rapid after release of the clamp, a picture characterized by congestion and dilatation of arterioles and capillaries. 8. The fact that accelerated respirations result from emboli in the pulmonary capillaries and arterioles only after a certain quantity of material has been introduced, and the fact that emboli in one lung do not occasion accelerated respirations unless the circulation through the other lung is occluded or abnormal, leads us to the conclusion that the phenomenon is not an irritative stimulus due to foreign bodies, but is in some manner related to (a) diminution of the pulmonary vascular bed, (b) resistance to the blood flow through the lungs or (c) congestion or dilatation of the arterioles and capillaries of the lungs.     

Reproducibility and validity of a food frequency questionnaire in European and polynesian New Zealanders

The reproducibility and validity of a self‐administered 142‐item food‐frequency questionnaire (FFQ) was assessed in a population comprising 124 European and 52 Polynesian (17 Maori and 35 Pacific Island) New Zealanders aged 40–65 years. Reproducibility correlation coefficients, determined by administration of the same questionnaire on two occasions 3 years apart, were higher in European than Maori and Pacific Island. participants, ranging from 0.47 to 0.87 in Europeans (median 0.66) and from 0.41 to 0.79 in Maori and Pacific Island people (median 0.44). In general, there were no significant differences in mean nutrient intakes calculated from the two FFQs by Europeans or Maori and Pacific Island participants despite their cultural and language differences. When the FFQ was compared with a 3‐day food diary in a sub sample of 101 Europeans, 15 Maori and 22 Pacific Islanders, the validity was good for most nutrients, with overestimation of a few nutrients in each ethnic group. Correlation coefficients between the 3‐day food diary and FFQ ranged from 0.41 to 0.81 in Europeans (median 0.48) and from 0.36 to 0.56 in Maori and Pacific Island people (median 0.55). Ratios of energy intake to resting metabolic rate suggested that Maori and Pacific Island people were more likely to underestimate their habitual energy intake by the 3‐day diet diary method compared to Europeans, but that Europeans were more likely to underestimate total energy intake by the food frequency method and Pacific Island participants to overestimate it. Obese Europeans and Maori were more likely to under‐report dietary intakes by the 3‐day diary method. We conclude that our FFQ performed better in European than Maori and Pacific Island participants.