A melon pulp concentrate rich in superoxide dismutase reduces stress proteins along the gastrointestinal tract of pigs.

OBJECTIVE: A melon (Cucumis melo LC.) pulp concentrate (MPC) rich in superoxide dismutase (SOD) activity was tested for its ability to decrease stress protein expressions along the gastrointestinal tract in a swine model. METHODS: Pig sextuplets weaned at 21 d of age were selected from among six litters (n = 36). After a 2-d fasting period, the pigs were fed at similar levels of intake of the control, MPC1, and MPC2 diets, which provided 0, 12.5, and 50 IU of added SOD per kilogram of food, respectively. One triplet of pigs per litter was slaughtered at 7 d and the second triplet at 14 d after weaning. SOD, catalase, and digestive enzymes were determined enzymatically and stress protein expressions by western blotting. RESULTS: Plasma SOD increased with MPC dose at day 14 (P < 0.05). Mucosal weights in the proximal and mid small intestine were lower at day 14 (P < 0.05), cecum tissue weight was greater (P < 0.05), and sucrase-specific activity in mid and distal small intestine mucosa was lower (P = 0.05) in the MPC2 group than in the control group. MPC supplementation essentially decreased (P < 0.05 to P < 0.001) stress proteins in the stomach (all), the mid small intestine (heat-shock protein-27, neuronal nitric oxide synthase) and the colon (heat-shock protein-70, neuronal nitric oxide synthase). CONCLUSION: A SOD-rich MPC provided at the dose of 50 IU/kg of food for up to 12 d was effective in lowering the level of stress proteins along the gastrointestinal tract of pigs after weaning.

Consumption of fumonisin B1 for 9 days induces stress proteins along the gastrointestinal tract of pigs.

Fumonisin B(1) (FB1) is a mycotoxin which alters intestinal epithelial cell physiology and barrier properties, and accumulates in the colon. Data on effects of FB1 on stress proteins in the gastrointestinal tract (GIT) are lacking. Therefore, we hypothesized that repeated consumption of FB1 alters GIT tissue levels of stress proteins. This was tested using 36 weaned pigs fed a FB1 solution (n=18) or the vehicle (control; n=18) for 9 days. The pigs were then slaughtered, the organs were weighed and GIT tissues were collected for assessing GIT integrity, and for analysing stress proteins by Western blotting and densitometry (n=7 in each group). FB1 had little effects on growth rate but the liver was heavier (P<0.01) in FB1-fed pigs. alphaB crystallin and COX-1 concentrations were eight-fold and 12-fold higher in the colon of FB1-fed pigs than in the controls (P<0.0001). Concentrations of COX-1 and nNOS in the stomach, HSP 70 in the jejunum and HO-2 in the colon were also higher in FB1-fed pigs (P<0.05 to P<0.001). In conclusion, the FB1 extract drastically enhanced colonic levels of alphaB crystallin and COX-1, with milder increases in other stress proteins along the GIT of pigs. The data suggest that the colon is an important target for FB1-induced stress responses.

Comparative assessing for radiological, chemical, and physical exposures at the French uranium conversion plant: Is uranium the only stressor?

This study presents the pattern of exposure to uranium and other occupational pollutants known to be potentially carcinogenic, mutagenic or toxic and used at the main uranium conversion plant in France. For different uranium compounds specified according to their solubility and purity, and 16 other categories of pollutants: chemicals, fibres, vapours, dust, and heat a time- and plant-specific job exposure matrix (JEM) was created covering the period 1960-2006. For 73 jobs and for each pollutant the amount and frequency of exposure were assessed on a four-level scale by different time periods. The JEM shows 73% sensitivity and 83% specificity. Although exposure assessment was semi-quantitative, the JEM allows computing of individual cumulative exposure score for each pollutant across time. Despite the predominant natural uranium compounds exposure, the amount of exposure to other pollutants such as TCE and other chlorinated products, asbestos, and fibres, is important at the plant. Numerous correlations detected between uranium compounds exposure and exposure to chemicals warrants improving biological monitoring of exposed workers and accounting for associated exposures in epidemiological studies. Results of this study will be used for further investigation of association between exposure and mortality among uranium conversion workers cohort.

Up-regulation of heat shock protein HSP 20 in the hippocampus as an early response to hypoxia of the newborn.

Hypoxia is an important challenge for newborn mammals. Stress generated at the brain level under low oxygenation conditions results in up-regulation of heat shock proteins (HSPs) and other stress proteins. The aim of the present work was to determine the effect of hypoxia in the newborn on some newly described small molecular weight HSPs (HSP 20 and B8) in the hippocampus, cortex and cerebellum of newborn piglets. These effects will be compared with those of other closely related proteins such as alphaB crystallin, HSP 27, heme oxygenase (HO)-1, HO-2, cyclooxygenase (COX)-1 and COX-2. The piglets were submitted to hypoxia (5% O(2); 95% N(2)) over either 1 or 4 h, with recovery periods ranging from 0 to 68 h. Western blot analysis showed that HSP 20 was rapidly induced only in the hippocampus, long before hypoxia-inducible transcription factor HIF-1alpha, while HSP 27 was rapidly induced in the cortex and cerebellum. Vascular epithelial growth factor was increased simultaneously in the three regions. Moreover, an increase in the expression of, respectively, HO-1 and COX-2 was observed later, but at the same time, in the three regions tested. It appears that HSP 20 can be an early marker of hypoxia in the hippocampus. The other small HSPs or stress proteins display different temporal patterns of up-regulation (HSP 27 and HO-1, COX-2) or do not show changes in their expressions (alphaB crystallin, HSP B8, HO-2 and COX-1).

Overexpression of alphaB crystallin in the gastrointestinal tract of the newborn piglet after hypoxia.

Newborn animals are particularly sensitive to hypoxic stress. Oxygen is spared for sensitive tissues, including brain and heart. Scarce information is available concerning the molecular effects of hypoxia in the gastrointestinal tract (GIT). Moreover, stress protein expressions, including heat shock proteins (HSP), are still poorly documented in the GIT. Our objective was to determine the possible effect of hypoxia on HSP expression at birth. After western blotting, alphaB crystallin, HSP 27, and HSP 70 expressions were determined in newborn controls and piglets exposed to 1 or 4 h hypoxia (5% O2, 95% N2) allowed to recover from 1 to 68 h. Cytosol and nuclei were also separated and the extracts were tested for HSF1 and alphaB crystallin expressions. Surprisingly, alphaB crystallin was overexpressed in the stomach and colon in animals submitted to hypoxia, whereas HSP 27 and HSP 70 expression remained stable. Increases and return to basal levels in HSF1 and alphaB crystallin were simultaneously observed in the unique nuclear compartment. To our knowledge, the present study is the first to demonstrate the oxygen dependency of an HSP in the GIT, particularly in the colon in newborn piglets. The kinetics of alphaB crystallin overexpression after hypoxia parallels the activation of HSF1. This observation possibly indicates a correlation between this factor and alphaB crystallin after hypoxia. Taken together, the present results open the field of wide investigation about the specific response of this low-molecular-weight HSP and its possible involvement in pathological states in the GIT such as stomach and colon.

Effects of hypoxia on stress proteins in the piglet brain at birth.

Newborn piglets were submitted to normobaric hypoxia (5% O2, 95% N2) for either 1 or 4 h. The effects of hypoxia on the neonatal brain were characterized through a time-course analysis of levels of various proteins such as heat shock proteins (HSP27, 70, and 90), hypoxia inducible factor-1alpha (HIF-1alpha), neuronal nitric oxide synthase (nNOS), hemeoxygenase-2 (HO-2), and caspase-3. The expression of these proteins was determined at different stages of recovery up to 72 h in cerebellum, cortex, and hippocampus by Western blot analysis in hypoxic maintained animals that were made hypoxic at either 20 or 37 degrees C. In all regions of the brain, HIF-1alpha and HSP27 expression were strongly increased until 22 h of recovery. No significant changes were observed for HSP70, HSP90, and HO-2. A small elevation of expression of nNOS was observed at early stages in the cerebellum and the cortex with no change in the hippocampus. Expression of caspase 3 was strongly increased in the cortex 24 and 48 h after hypoxia but unchanged in the hippocampus. These results are presented in terms of the porcine model of nonischemic hypoxia and its delayed neuronal effects on the cerebral outcome. Because of their recently established biochemical and functional interactions, the expression of the main HSPs, HIF-1alpha, nNOS, and caspase-3 after hypoxia are delineated.

Expression of small heat shock proteins HspB2, HspB8, Hsp20 and cvHsp in different tissues of the perinatal developing pig.

Recently, we have described the developmental expression of the small heat shock proteins (sHsps) Hsp27/HspB1 and alphaB-crystallin/HspB5 in different tissues of pigs from almost full-term foetuses to three years old adults (P. Tallot, J. F. Grongnet, J. C. David, Biol. Neonate, 83, 281-288, 2003). The data described in this report extends this study to four other members of the sHsp family (Hsp20/HspB6, cvHsp/HspB7, MKBP/HspB2 and HspB8). We studied expression of these proteins in porcine lens, brain, heart, liver, kidney, lung, skeletal muscle, stomach, and colon, and found a ubiquitous expression of Hsp20 and HspB8 as earlier reported for Hsp27 and alphaB-crystallin. In contrast, cvHsp and HspB2 expression is essentially restricted to heart and muscle. During development, the sHsps tend to (temporarily) increase in stomach, liver, lung, kidney, hippocampus, and striatum, while expression in heart is more or less constant, and a large variation is found in sHsp expression patterns in skeletal muscle. In cerebellum and cortex a temporary decrease of Hsp20 and HspB8 is observed directly after birth. The major impact of this study is that each tissue seems to have a unique profile of sHsp expression, which varies during development and may reflect the need of a particular tissue to maintain at all stages an optimal chaperoning machinery to protect against physiological stress.

Reciprocal variations of nNOS and HSP90 are associated with fasting in gastrointestinal tract of the piglet.

The effects of fasting on neuronal NO synthase (nNOS), Heme oxygenase 2 (HO-2), and heat shock protein 90 (HSP90) was determined by immunoblotting in the stomach, duodenum, mid-jejunum, distal ileum, and proximal colon of 28-day-old piglets. nNOS expression was drastically reduced in all the gastrointestinal areas studied while HO-2 was not changed. Concomitant with the nNOS decrease, elevated expressions of HSP90 were observed in these different areas. These results are discussed in terms of the regulation relationship between NOS and HSP90 and the possible protective effect of the heat shock protein and the potential application in digestive pathologies.

Weaning affects the expression of heat shock proteins in different regions of the gastrointestinal tract of piglets.

Heat shock proteins (HSP) play a central role in the protection of cells, tissues or organs subjected to various types of stressors. Different nutrients have been recently shown to exert their protection through the induction of HSP. Because these nutrients alleviate alterations of the intestine after weaning in pigs, this study was designed to obtain basic information on the expression of HSP 27, heat shock cognate 70 (HSC 70), HSP 70 and HSP 90 along the gastrointestinal tract (GIT) of young pigs and to study the effect of weaning on this expression. Pigs were weaned at 28 or 21 d and slaughtered at various times postweaning. All HSP were expressed in the GIT segments studied before and after weaning. However, the expression of HSP 27 and HSP 70 was transiently increased in the stomach and duodenum between 6 and 12 h postweaning and between 24 and 48 h in the mid-jejunum, ileum and colon. Their expressions were transiently decreased in the ileum. Expression of HSP 90 increased in the stomach and jejunum but decreased in the duodenum, ileum and colon. Similar results were obtained at both ages of weaning. We conclude that the HSP studied are present all along the gut of pigs and that their expression is modulated through weaning according to spatial-temporal patterns. The modulation by nutrients of HSP and their protective role on the GIT remain to be investigated in pigs.