Modulation of adipokines and cytokines in gestational diabetes and macrosomia.

CONTEXT/OBJECTIVE: Not much is known about the implication of adipokines and different cytokines in gestational diabetes mellitus (GDM) and macrosomia. The purpose of this study was to assess the profile of these hormones and cytokines in macrosomic babies, born to gestational diabetic women. DESIGN/SUBJECTS: A total of 59 women (age, 19-42 yr) suffering from GDM with their macrosomic babies (4.35 +/- 0.06 kg) and 60 healthy age-matched pregnant women and their newborns (3.22 +/- 0.08 kg) were selected. METHODS: Serum adipokines (adiponectin and leptin) were quantified using an obesity-related multiple ELISA microarray kit. The concentrations of serum cytokines were determined by ELISA. RESULTS: Serum adiponectin levels were decreased, whereas the concentrations of leptin, inflammatory cytokines, such as IL-6 and TNF-alpha, were significantly increased in gestational diabetic mothers compared with control women. The levels of these adipocytokines were diminished in macrosomic babies in comparison with their age-matched control newborns. Serum concentrations of T helper type 1 (Th1) cytokines (IL-2 and interferon-gamma) were decreased, whereas IL-10 levels were significantly enhanced in gestational diabetic mothers compared with control women. Macrosomic children exhibited high levels of Th1 cytokines and low levels of IL-10 compared with control infants. Serum IL-4 levels were not altered between gestational diabetic mothers and control mothers or the macrosomic babies and newborn control babies. CONCLUSIONS: GDM is linked to the down-regulation of adiponectin along with Th1 cytokines and up-regulation of leptin and inflammatory cytokines. Macrosomia was associated with the up-regulation of Th1 cytokines and the down-regulation of the obesity-related agents (IL-6 and TNF-alpha, leptin, and adiponectin).

N-3 fatty acids modulate antioxidant status in diabetic rats and their macrosomic offspring.

OBJECTIVE: We investigated the role of dietary n-3 polyunsaturated fatty acids (n-3 PUFA) in the modulation of total antioxidant status in streptozotocin (STZ)-induced diabetic rats and their macrosomic offspring. DESIGN: Female wistar rats, fed on control diet or n-3 PUFA diet, were rendered diabetic by administration of five mild doses of STZ on day 5 and were killed on days 12 and 21 of gestation. The macrosomic (MAC) pups were killed at the age of 60 and 90 days. MEASUREMENTS: Lipid peroxidation was measured as the concentrations of plasma thiobarbituric acid reactive substances (TBARS), and the total antioxidant status was determined by measuring (i) plasma oxygen radical absorbance capacity (ORAC), (ii) plasma vitamin A, E and C concentrations, and (iii) antioxidant enzymes activities in erythrocytes. The plasma lipid concentrations and fatty acid composition were also determined. RESULTS: Diabetes increased plasma triglyceride and cholesterol concentrations, whereas macrosomia was associated with enhanced plasma cholesterol and triglyceride levels, which diminished by feeding n-3 PUFA diet. N-3 PUFA diet also reduced increased plasma TBARS and corrected the decreased ORAC values in diabetic rats and their macrosomic offspring. EPAX diet increased the diminished vitamin A levels in diabetic mothers and vitamin C concentrations in macrosomic pups. Also, this diet improved the decreased erythrocyte superoxide dismutase and glutathione peroxidase activities in diabetic and macrosomic animals. CONCLUSION: Diabetes and macrosomia were associated with altered lipid metabolism, antioxidant enzyme activities and vitamin concentrations. N-3 PUFA diet improved hyperlipidemia and restored antioxidant status in diabetic dams and MAC offspring.

Antioxidant status in alcohol-related diabetes mellitus in Beninese subjects.

In the present study, we investigated the antioxidant status in diabetes mellitus, related or not to alcohol consumption. A total of 38 type 1, 48 type 2 and 42 alcohol-related diabetic patients were selected. Total antioxidant status was assessed through the oxygen radical absorbance capacity of the plasma and the determination of enzymatic and non-enzymatic antioxidant molecules. Serum triglycerides, total cholesterol and HDL-cholesterol concentrations were determined and the lipid peroxydation was evaluated by measuring thiobarbituric acid reactive substances (TBARS) assay. Plasma total antioxidant capacity was more decreased in alcohol-related diabetes than that in type 1 and type 2 diabetes, regardless of the complications (retinopathy and renal failure). Plasma vitamin E concentrations were significantly decreased whereas those of vitamin C increased in all of the diabetic patients compared to the controls, irrespective to the complications. In addition, superoxide dismutase and glutathione peroxidase activities were reduced in all the patients (type 1, type 2 and alcohol-related), irrespective to the complications. Glutathione reductase activity was diminished in type 1 and alcohol-related, but not in type 2, diabetic patients. Glutathione (GSH) concentrations significantly decreased in all diabetic patients with a significant decrease in alcohol-related diabetic patients. Excessive alcohol consumption appears as an oxidative aggravating factor in diabetes mellitus. Besides, alcohol-related diabetes highly resembles to type 1 diabetes as far as the antioxidant parameters are concerned.

In vitro effects of four tropical plants on the activity and development of the parasitic nematode, Trichostrongylus colubriformis.

The in vitro effects of extracts of four tropical plants (Zanthoxylum zanthoxyloides, Newbouldia laevis, Morinda lucida and Carica papaya) on the egg, infective larvae and adult worms of Trichostrongylus colubriformis were screened for potential anthelmintic properties. Significant effects were observed with the four plants on T. colubriformis but they differed depending on the stage of the parasite. Extracts of each plant induced a dose-dependent inhibition of egg hatching. Using a larval inhibition migration test, the effects on the infective larvae were also detected with the four plant extracts. In contrast, for adult worms, the effects were statistically significant only for N. laevis and C. papaya. No significant activity was shown for M. lucida and Z. zanthoxyloides. These in vitro results suggest the presence of some anthelmintic properties associated with these four plants, which are traditionally used by small farmers in western Africa. These effects need to be studied under in vivo conditions.

In vivo effects of Fagara leaves on sheep infected with gastrointestinal nematodes.

Infections of the gastrointestinal tract with parasitic nematodes, and particularly Haemonchus contortus, represent one of the main pathological constraints on the breeding of small ruminants in tropical countries. Owing to the constant development of anthelmintic resistance in worm populations, alternative control methods are being evaluated, including an assessment of the value of the traditional plants. The effect of the consumption of Fagara leaves on sheep infected with gastrointestinal nematodes was therefore measured both under controlled conditions and in a field survey. In sheep experimentally infected with 2500 third-stage larvae of H. contortus, a three-day administration of Fagara was associated with a decrease in egg excretion and a significant reduction in the fertility of female worms, without changes in the worm number. A reduction in egg output was also confirmed from the results obtained of a survey or naturally infected sheep performed in smallholder farms. This survey also indicated that the repeated consumption of small amounts of Fagara leaves was more effective than a single three-day administration. Results of both studies suggest that Fagara leaves might be an alternative option to chemotherapy to modulate the epidemiology of H. contortus in tropical countries.

In vitro effects of four tropical plants on three life-cycle stages of the parasitic nematode, Haemonchus contortus.

Alcoholic extracts of four tropical plants (Zanthoxylum zanthoxyloides, Newbouldia laevis, Morinda lucida and Carica papaya) were screened in vitro for potential anti-parasitic effects against eggs, infective larvae and adult Haemonchus contortus. Significant effects were obtained with all four plants but differences were observed depending on the parasitic stage. The effects of the four plant extracts were similar on egg hatching and were dose dependent. In contrast, no dose-response relationship was found for infective larvae and adult worms, although more potent effects were usually observed with the highest concentrations. Using a larval inhibition migration test, extracts of fagara (Z. zanthoxyloides) were found to be less active against Haemonchus infective larvae than were the other plants. N. laevis was found to be highly and rapidly effective against adult worms. Overall, these in vitro results suggest that these four plants, traditionally used by small farmers in Western Africa, do possess anti-parasitic properties. These effects remain to be confirmed through in vivo studies.

Antioxidant status and levels of different vitamins determined by high performance liquid chromatography in diabetic subjects with multiple complications.

Plasma vitamin A, C and E levels and erythrocyte antioxidant enzyme activities were investigated in type I and type II diabetic subjects with and without complications, i.e., hypertension, coronary artery disease and renal failure. Reverse phase HPLC was used to quantify vitamin A and E levels. We observed that the vitamin C levels were not significantly different between control and diabetic subjects. However, vitamin A and E levels were significantly lower in type I and type II diabetic subjects compared to controls. Superoxide dismutase (SOD) activity was significantly lower in type II, but not in type I, diabetic patients compared to controls. Interestingly, glutathione reductase and peroxidase activities were diminished in type I, but not in type II, diabetic subjects as compared to controls. Catalase activity was lower in both types of diabetic patients in comparison with their respective controls. Altogether these results suggest that diabetes mellitus may be associated with altered antioxidant status regardless to various complications.

Spermatogenesis in the grasscutter, Thryonomis swinderianus (Rodentia).

Spermatogenesis in the grasscutter, Thryonomis swinderianus, is characterized by remarkable diversification of the endoplasmic reticulum. In spermatocytes, besides the usual form with flat cisternae and vesicles, this membrane system forms unusually wide cisternae and vacuoles containing moderately electron dense material near the Golgi apparatus and the cell membrane. In spermatids a whorl of smooth cisternae enclosing aggregates of granular electron dense material appears as another specialization. Although all parts of the endoplasmic reticulum are in connection with each other, their distinct structural differences suggest functionally different subcompartments. Once established, the structures persist even after cytoplasm has been discarded from the nearly mature sperm. In the elongating spermatid a thin plica of the Sertoli cell begins to separate the developing structures from the remainder of the cytoplasm containing the endomembrane systems, part of which is further ensheathed when this fold enroles to form an incomplete tube. Occasionally, in late spermatids a spindle-shaped body of paracrystalline structure occurs temporarily around the flagellum at the beginning of the main piece.

Calcium binding proteins immunohistochemistry and identification of neurons in the mammalian pineal gland of the African giant rat: Cricetomys gambianus.

The presence of true neurons in the rodent pineal gland is still a matter of controversy. In this work, by using immunohistochemistry with five antibodies against calcium-binding proteins (calbindin-D28k, calretinin, calmodulin, neurocalcin and S-100 beta) and Cricetomys gambianus, a rodent belonging to Muridae family living in Africa, we were able to illustrate the presence of neurons in the pineal gland. Anti-calbindin-D28k and anti-calretinin labelled neurons belonging to two neural ganglia. One ganglion was localized in the anterior part of the gland near the pineal stalk and the other one in the posterior portion of the organ. Immunoreactive neurons are medium in size (15-20 microns) and have long thick processes running towards the stalk. Calretinin and calbindin-D28k positive neurons stained with different intensities. Thin processes were detected by anti-calretinin whereas thick processes were preferentially calbindin-D28k positive. Neurocalcin labelled a few smaller neurons and many thin processes within the ganglion. Calmodulin could not be detected immunochemically. Within the ganglia many astrocytic processes were S-100 beta positive. The afferent and the efferent pathways of the pineal ganglia remain to be elucidated.

Interstitial tissue in the testis of the giant rat, Cricetomys gambianus.

Besides Leydig cells macrophages form considerable part of the interstitial tissue in the giant rat, Cricetomys gambianus. Leydig cells contain bundles of usually six to twelve parallel tubes with walls of longitudinal layers alternating with circular layers of microfilaments. Polyribosomes, both free and membrane bound, are concentrated around or inside the tubes. The tubes may enclose mitochondria and microperoxisomes as well. The light cytosol of the macrophages is dotted with electron-dense 5-nm particles which might be ferritin. Specific contacts exist between Leydig cells and macrophages indicating a functional interdependence between these two cell types.

Absence of short-wavelength sensitive cones in the retinae of seals (Carnivora) and African giant rats (Rodentia).

Most non-primate mammals have two types of cone: short-wavelength sensitive (S) and middle-to-long-wavelength sensitive (M/L) cones. In two species of African giant rats, Cricetomys gambianus and C. emini, and in two species of earless seals, Phoca hispida and P. vitulina, the retinal cone types and cone distributions were assessed with antibodies specific for the M/L-cone opsin and the S-cone opsin, respectively. All four species were found to completely lack S-cones, while M/L-cones were present in low densities. M/L-cone densities, rod densities and cone/rod ratios were determined across the retina. Cone proportions are about 0.3-0. 5% in C. gambianus, 0.5-0.8% in C. emini, and 1.5-1.8% in P. hispida. An absence of S-cones has previously been reported in a few nocturnal mammals. As earless seals are visually active during night and day, we conclude that an absence of S-cones is not exclusively associated with nocturnality. The functional and comparative aspects are discussed.

Calbindin localization in African giant rat kidney (Cricetomys gambianus).

Cricetomys gambianus are rodents living in savanna and follow area. They can live with restricted drinking water eating fresh food. Therefore their kidney may have some adaptive mechanisms for ion/water homeostasis compared to usual laboratory rats. In this study we have looked for calbindin, an intracellular calcium binding protein previously found in distal convoluted tubules from all mammalian species that have been studied and able to increase, in vitro, Ca2+ reabsorption. We have shown by using in situ hybridization, immunoblotting and immunohistochemistry that calbindin was expressed in three different portions of the distal nephron of the African giant rat. Calbindin was found in distal convoluted tubules, in cortical collecting tubules and in outer medullary collecting ducts. By contrast, in laboratory rat, calbindin was only found in distal convoluted tubules and undetectable in medullary collecting ducts. Thick ascending limb of Henle's loop were calbindin negative as shown by double immunolabelling using anti-uromucoid (Tamm-Horsfall protein). As previously shown in laboratory rat and rabbit, transcellular Ca2+ movement seems to be facilitated by calbindin in renal tubules segments predominantly actively transporting Ca2+, it may be suggested that in African giant rat, outer medullary collecting ducts may also actively transport Ca2+. As calretinin, another intracellular calcium binding protein highly homologous to calbindin but whose function is still conjectural has been suspected to be expressed in kidney, we have looked and not found any calretinin in both adult rat species.

Paneth cells of African giant rats (Cricetomys gambianus).

The ultrastructure of Paneth cells of African giant rats (Cricetomys gambianus), which were captured on the savanna in western Africa, was studied. The Paneth cells of Cricetomys were clustered at the bottom of crypts of the small intestine, but not of the colon. In the normal state, Paneth cells had a few secretory granules showing high electron density. Small clear vesicles which are a characteristic in laboratory albino rats were not conspicuous. Vacuolated Paneth cells and secreted materials from the Paneth cells were frequently found. This suggests that the secretion of Paneth cells of Cricetomys is active in the natural state. No phagocytotic figures were observed. After atropine sulfate treatment, secretory granules increased in size and number, whereas the electron density decreased, similar to that of goblet cell granules. However, the granules were not stained by alcian blue or by PAS. Inhibition of secretory stimuli by atropine can alter the intracellular processing of secretory substances in Paneth cells.

Epithelial cell migration on small intestinal villi in the neonatal rat. Comparison between [3H] thymidine and cytoplasmic labelling after Pu-citrate ingestion.

This study compares, in 2-d-old rats, the migration rates of epithelial cells on villi of the small intestine, using two labelling methods: a single [3H] thymidine injection; and cytoplasmic labelling by a single ingestion of Pu-citrate. Histoautoradiography showed negligible diffusion of Pu after the initial retention, which was mostly confined to the epithelial cells of the villi. However, after sloughing of labelled cells in the intestinal lumen, Pu was reabsorbed by the distal epithelial cells. In segments in which Pu reabsorption was negligible, the migration rates of Pu- and 3H-labelled cells were very close. These rates, expressed in micrometers, were almost constant along the length of the villus, and the Pu and 3H labelling edges reached the top of the villi in about 5 and 7 d, respectively. Once Pu retention had reached its maximum in 9 equal segments cut along the small intestine, tissue counting showed an exponential Pu release of 30-40%/d from each segment until the end of the experiment at d16. This constant release might reflect a constant cell migration rate during the period from Pu ingestion until d16.

Localization of plutonium retention in the small intestine of the neonatal rat, guinea pig, baboon and macaca after Pu-citrate ingestion.

The retention of Pu-citrate in the gastrointestinal wall was compared at similar post ingestion times after ingestion at 2 days of age by rats and guinea pigs and at 1 to 34 days by neonatal primates. The small intestine was the main site of the Pu retention in all species. In rats and primates, most of the Pu was retained in the distal ileum, whereas in guinea pigs it was more homogeneously distributed. In the rats, Pu was retained in the epithelial cells on villi, but in the guinea pigs and primates it was confined to the macrophages under the epithelial cells in the lacteal region.

Effects of the chemical forms and valency states of neptunium on its jejunal transfer in the rat.

The transfer of various Np(IV) and Np(V) chemical forms across the small intestine of rats was measured in instilled and perfused jejunum. Instillation of Np(V) nitrate together with citrate or DTPA resulted in the same absorption of Np as after instillation of Np(V) nitrate alone (3 per cent per hour). Perfusion of Np(V) nitrate with bicarbonate or DTPA resulted in a similar transfer (2 per cent) but added citrate or ascorbate resulted in reduced transfer (0.8 per cent). Addition of phytate reduced Np transfer in both instilled and perfused jejunum (0.4 per cent). Np(IV) transfer was usually the same as, or less than that of, the corresponding Np(V) forms. Np(IV) transfer was similar in perfused and instilled jejunum, increasing from 0.2 per cent in the presence of citrate and phytate, to 1 per cent with EDTA and DTPA. Except for phytate, all the forms of Np(V) tested behaved like Np(V) nitrate after transfer from the intestine or after intravenous injection. By contrast, the behaviour of Np(IV) varied for all the forms tested and, for a given form, varied as a function of the experimental procedure used, i.e. jejunal instillation, perfusion, or intravenous injection. These findings suggest that the intestinal transfer of Np might occur via the intercellular pathway, and that it is controlled by both the molecular weight of the Np compound and its stability constant.

Acute lesions induced by alpha-irradiation of intestine after plutonium gavage of neonatal rats.

After gavage of two-day-old rats with 238Pu(IV)-citrate at 17.4 MBq/kg (122 kBq per animal), 45 per cent of the animals died during the second week following ingestion. Histological analysis showed that death was due to acute intestinal lesions caused by alpha-radiation that resulted in denudation of the ileum. Under these experimental conditions, the total alpha-dose delivered to the ileal wall and its contents was estimated at 150 Gy. No acute lesions were observed after gavage of two-day-old rats with Pu-citrate at 5.3 MBq/kg.