Qualitative and quantitative ultrastructural analysis of the mitochondria from adrenal gland cortex under the action of viperidae family snake venoms / Análisis ultrastructural cualitativo y cuantitativo de la mitocondria de la glándula adrenal bajo la acción de los venenos de serpientes de la familia viperidae

SUMMARY: The Viperidae venoms are composed of a mixture of constituents with enzymatic and non-enzymatic actions, which act on ultrastructural components of cells and tissues. Here, the number of mitochondria, mitochondrial area and the number of mitochondrial cristae from adrenal glands cortex treated with snake venoms were tested after 3, 6 and 24 hours of venom injections. The mitochondria quantitative changes showed a statistically significant decrease, in the number of mitochondria past 3, 6 and 24 h. There was an increase in the mitochondrial area after 6 h, where Crotalus vegrandis venom did not present significant differences with Crotalus pifanorum or Bothrops venezuelensis venoms. After 24 h, there was an escalation of mitochondrial area in all tested venoms. The number of mitochondrial cristae after 3 h did not present important differences with the control treatment. After 6 h, the number of mitochondrial cristae initiated to decrease under the activities of the 3 venoms action, until 24 h of observation. In the qualitative observations it was possible to witness an intense damage of the mitochondria, with loss and swelling of membranes, disappearance of cristae and the appearance of myelin figures, which started at 3 h after the Crotalus and Bothrops venoms injections. These damages probably were due to cytotoxic effects of phospholipases, metalloproteases and/or other proteolytic activities present in Viperidae snake venoms, being more evident in Crotalus venoms. As far as we know, these results define a novel finding that suggest that Viperidae snake venoms are extremely toxic to mammalian mitochondria.

RESUMEN: Los venenos de Viperidae tienen acciones enzimáticas y no enzimáticas, que actúan sobre la estructura celular. Aquí se probaron, a las 3, 6 y 24 horas de la inyección del veneno, el número de mitocondrias, el área mitocondrial y el número de crestas mitocondriales de la corteza de las glándulas adrenales. Los cambios cuantitativos de las mitocondrias mostraron una disminución en el número de mitocondrias a las 3, 6 y 24 h. Hubo un aumento en el área mitocondrial a las 6 h, donde el veneno de la serpiente Crotalus vegrandis no presentó diferencias significativas con los venenos de Crotalus pifanorum o Bothrops venezuelensis. Después de 24 h, hubo un aumento del área mitocondrial en todos los venenos. El número de crestas mitocondriales a las 3 h no presentó alteraciones o diferencias importantes con el tratamiento de control. Después de 6 h, el número de crestas mitocondriales comenzó a disminuir bajo la acción de los 3 venenos, hasta las 24 h de observación. En las observaciones cualitativas se observó un daño intenso de las mitocondrias, con pérdida y edema de las membranas, desaparición de las cristae y aparición de figuras mielínicas, que comenzó a las 3 h después de las inyecciones de veneno de Crotalus y Bothrops. Estos daños se debieron factiblemente a los efectos citotóxicos de componentes proteolíticos de los venenos. Creemos que estos resultados definen un nuevo y original hallazgo, que sugiere que los venenos de serpiente Viperidae son extremadamente tóxicos para las mitocondrias de mamíferos.

Histopathological and immunohistochemical changes induced by Bisphenol A in reproductive tissues of female rat

Background: Bisphenol A [BPA] is an environmental chemical that has been widely used in the manufacture of polycarbonate plastics and epoxy resins for many years. Due to its major applications in the production of plastic food or beverage containers and the coating of food cans, people of different ages are inevitably exposed to BPA in daily life. It is a contaminant with increasing exposure to it and exerts both toxic and estrogenic effects on mammalian cells

Aim of the work: the present study was designated to evaluate the histopathological and immunohistochemical effect of BPA on the histoarchitecture of pituitary ,adrenal, ovarian and uterine axis of female albino rats and the ameliorative effect of antiestrogen drug and stem enhance

Experimental model and methods: 20 female albino rats weighing 100 - 120 g. were kept under observation for about 15 days before the onset of the experiment for adaptation, then the rats were classified into 4 groups 5 rats for each , the first group was left without any treatment for 30 days as negative control group , the second group was administered with 20 mg/kg.bw of BPA for 15 consecutive days as positive control, the third group administered with 20 mg/kg.bw of BPA for 15 consecutive days and then treated with antiestrogen drug as 0.1 mg/100gm.bw for 15 day, the fourth group administered with the same dose for the same period and the treated with stem enhance [4.5 mg/100.bw] for 15 days. All rats are scarified and organs were histologically examined after processing

Results: The results showed that PA has a histopathological effects on vital organs [pituitary, adrenal, ovary, oviduct and uterus] even for a short period with minimal ameliorative effect of antiestrogen drug and stem enhance

Stress modulating antioxidant effect of Nardostachys jatamansi.

The rhizomes of Nardostachysjatamansi, the plant commonly known as Jatamansi have been described in Ayurveda for their soothing and sedative action on the central nervous system. In the present study, the anti-stress effect of hydroethanolic extract (70%) of N. jatamansi (NJE) was evaluated in reference to its antioxidant property. Wistar rats were divided into four groups: naive, stressed, and T-200 and T-500 stressed with oral pre-treatment of NJE 200 and 500 mg/kg, respectively. Restraint of rats in metallic chambers for 4 h at 4 degreesC was followed by sacrifice and assessment of stress-induced alterations in biochemical parameters, incidence and severity of ulcers. Lipid peroxidation (LPO) and NO levels in stomach and LPO, NO levels and catalase activity in brain, plasma corticosterone level and adrenal ascorbic acid were measured. In vitro antioxidant activity of NJE was studied by measuring the free radical scavenging activity. NJE showed potent antioxidant activity and significantly reversed the stress-induced elevation of LPO and NO levels and decrease in catalase activity in the brain. It inhibited the incidence of gastric ulcerations and reversed the alterations in biochemical parameters/markers of stress-induced gastric ulceration. NJE also significantly altered stress-induced increase in adrenal and spleen weights and decrease in level of ascorbic acid in adrenal gland. Elevation of plasma corticosterone level was negated dose- dependently. The findings suggest that the NJE possesses significant anti-stress activity, which may be due to its antioxidant activity.

La melatonina reduce la respuesta de cortisol al ACTH en humanos / Melatonin reduces cortisol response to ACTH in humans

Background: Melatonin receptors are widely distributed in human tissues but they have not been reported in human adrenal gland. Aim: To assess if the human adrenal gland expresses melatonin receptors and if melatonin affeets cortisol response to ACTH in dexamethasone suppressed volunteers. Material and methods: Adrenal glands were obtained from 4 patients undergoing unilateral nephrectomy-adrenalectomy for renal cáncer. Expression of mRNA MT1 and MT2 melatonin receptors was measured by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). The effect of melatonin on the response to intravenous (i.v.) ACTH was tested (randomized cross-over, double-blind, placebo-controlled tríal) in eight young healthy males pretreated with dexamethasone (1 mg) at 23:00 h. On the next day at 08:00 h, an i.v. Une was inserted, at 08:30 h, and after a blood sample, subjeets ingested 6 mg melatonin or placebo. At 09:00 h, 1-24 ACTH (Cortrosyn, 1µg/1.73 m² body surface área) was injected, drawing samples at 0, 15, 30, 45 and 60 minutes after. Melatonin, cortisol, cortisone, progesterone, aldosterone, DHEA-S, testosterone and prolactin were measured by immunoassay. Results: The four adrenal glands expressed only MT1 receptor mRNA. Melatonin ingestión reduced the cortisol response to ACTH from 14.6+1.45µg/dl at 60 min in the placebo group to 10.8+1.2µg/dl in the melatonin group (p <0.01 mixed model test). It did not affect other steroid hormone levels and abolished the morningphysiological decline of prolactin. Conclusions: The expression ofMTl melatonin receptor in the human adrenal, and the melatonin reduction of ACTH-stimulated cortisol production suggest a direct melatonin action on the adrenal gland .

Strategies for minimizing corticosteroid toxicity: a review.

Glucocorticoids (GCs) are used commonly for the treatment of various pediatric inflammatory and autoimmune diseases. Although potent and generally effective, they are not without risks for producing serious adverse effects, especially when used in high doses for prolonged periods of time. For proper use of systemic glucocortcoids, a basic knowledge of the pharmacology, clinical usage guidelines, and adverse reactions of these agents is imperative. This review article emphasis on the commonly observed side-effects encountered with GC use in children and their underlying basic pathophysiological mechanisms. The appropriate anticipation of these side-effects with timely implementation of the suggested evidencebased guidelines has the potential significantly to prevent, minimize and treat common and disabling complications of glucocortcoid therapy.

Anti-stress and anti-oxidant effects of roots of Chlorophytum borivilianum (Santa Pau & Fernandes).

The aqueous extract of C. borivilianum (250 mg/kg for 7 days) significantly reverted the elevated levels of plasma glucose, triglycerides, cholesterol and serum corticosterone and also reduced the ulcer index, adrenal gland weight more as effectively as the standard drug (diazepam) in rats. At 125 mg/kg po, it showed a mild anti-stress activity. Under in vitro 1,1-diphenyl-2-picrylhydrazyl (DPPH*) free radical scavenging assay and lipid peroxidation assay the extract considerably inhibited, in a dose-dependent manner, the levels of DPPH* free radicals and thiobarbituric acid reactive substances, respectively thus showing significant antioxidant property. The results suggested that it could be used for the treatment of oxidative stress-induced disorders.

Multiple courses of antenatal steroids.

The benefits and risks of multiple courses of antenatal steroids (ANS) are still unresolved issues. This was a prospective cohort study in a level III neonatal unit. Preterm babies < or = 35 wk gestation were included. Malformations, chronic maternal steroid intake, exchange transfusions prior to cortisol sampling and incomplete ANS courses were exclusion criteria. Subjects were classified into: No course (Group 0), 1 course (group 1), 2 courses (Group 2), > 2 courses (Group 3) of antenatal dexamethasone. The key outcome was adrenal function assessed by basal and post-ACTH cortisol on day 3. Other outcomes were neonatal morbidity, mortality, growth parameters at birth, long term growth and neuro-development. Of 210 eligible babies, 124 were enrolled. 38, 51, 10 and 25 babies belonged to groups 0, 1, 2 and 3 respectively. Basal and post-ACTH serum cortisol did not show any significant difference between groups (p=0.5 and p=0.9 respectively). Incidence of severe HMD requiring ventilation was significantly lower (p=0.02) in multiple course group (combined groups 2 and 3) compared to single course group. There were no differences in other neonatal morbidity, birth OFC and weight between single and multiple ANS groups. Follow up data at a mean age of 22 mth was available in 59 subjects (69%) belonging to groups 1-3. No differences were noted in the proportion of patients with abnormal neurological examination (p=0.1), abnormal PDI (p=0.9), abnormal MDI (p=0.9) and physical growth between multiple and single course groups. Multiple courses of antenatal dexamethasone resulted in a significant decrease in severe forms of RDS and they did not cause adrenal suppression, decreased growth or impaired neuro-development.

Modulation of serum cortisol by substance P in albino rats: evidence of a direct effect on adrenal gland.

Effect of prolonged administration of substance P on the plasma cortisol level in the albino rats has been investigated. An inhibitory impact on intact individuals and a stimulatory effect in pharmacologically annulled rats has been observed. It is concluded that in normal conditions substance P presumably acts as a preventive agent for any excess secretion of cortisol while during stress or disturbed HPA or RAS conditions, it stimulates the secretion of cortisol. An intraglandular modulatory role of substance P has been suggested.

Effect of ovariectomy and of estrogen treatment on the adrenal gland and body weight in rats

To determine the effect of estrogen on adrenal gland histology and to ascertain whether or not estrogen regulates body and adrenal gland wet weight gain in the ovariectomized rat model. Two experiments investigated the effects of ovariectomy and estrogen replacement on the body and adrenal gland weight. We used 36 female Wistar Albino rats in this study. The study took placed in the Department of Medical Science Application and Research Center of Dicle University, Diyarbakir, Turkey, in 2002. Group 1 [control group] received no ovariectomy; all animals in the other groups were bilaterally ovariectomized and kept for 60 days. We designated group 2 and 3 as sham-operated group and bilaterally ovariectomized then in addition, group 3 received estradiol. We then cut the paraffin sections, which we obtained by routine histologic methods, into 5 mm and stained them with hematoxylin-eosin. We later evaluated the stained sections under a light microscope. The body weight was higher in the ovariectomized rats compared to the control groups. Ovariectomy did not result in significant changes in the wet weight of the adrenal gland. Furthermore, body weight increased after estrogen therapy, and the wet weight of the adrenal gland increased in the estrogen group. It was obtained in the sham-operated group that ovariectomy resulted in a decrease in the activity of the adrenal cortex. In the sham-operated and estrogen group, we observed a prominent capsule, expanded zona glomerulosa cells, regular parallel columns in zona fasciculata and there was an increased vascularization in the medulla, after the estrogen treatment. We determined that bilateral ovariectomy can result in a decrease in the activity of the adrenal cortex. In contrast, estradiol injection can cause a significant increase in the activity of the adrenal cortex and medulla

Effects of carbimazole on morphology of rat adrenals

Study was conducted to investigate the effects of [i] carbimazole and [ii] simultaneous administration of thyroxine [thyroid hormone analogue] on the morphology of rat adrenals. Design: A prospective case-control study. Place and Duration of Study: Department of Anatomy, Basic Medical Sciences Institute [BMSI], JPMC, Karachi, during a peroid of six weeks. Methology: The study was conducted on albino rats originally obtained from Charles River Breeding Laboratories, Brooklyn, Massachusetts, USA, and were cross bred, housed and maintained on balanced diet in the Animal House of BMSI, JPMC, Karachi.Eighteen adult albino rats used in the experiment were divided into three groups, i.e. A, B and C, each comprising 6 animals as control, carbimazole-treated, and carbimazole plus thyroxine-treated animals respectively. At the end of six weeks treatment all the animals were sacrificed and their adrenal glands removed, fixed, sectioned and stained with H and E and Sudan black. The histology of adrenals, width of cortex and its zones, and number of cells in different zones of cortex was studied. Carbimazole affects the morphology of adult rat adrenals by decreasing the total width of cortex and its zones especially the zona fasciculata with decrease in number of cells. It also showed the increase in fat contents on Sudan black staining. Carbimazole causes shrinkage of the adrenal cortex

Adrenocortical reserve and morphology in tuberculosis.

Ninety seven patients (63 males, mean age 31.8 years, SD 2.3) with various forms of tuberculosis were studied. All of them were HIV negative. Thirty normal control subjects (16 males, mean age 36.4 years, SD 1.8) were also studied. Fifty-eight of the 97 patients (59.8%) were malnourished (BMI < 18 kg/m2). The mean basal serum cortisol was lower in the TB group (n = 91) (351 nmol/1; SD 150) as compared to the normal control group (n = 8) (402 nmol/1; SD 93) but this difference did not attain statistical significance. Following administration of synthetic ACTH (cosyntropin), the 30 and 60 minutes mean serum cortisol values in the TB group were significantly lower as compared to the normal control group (p < 0.05). Forty five of the 91 patients (49.5%) who underwent the ACTH stimulation test had compromised adrenal reserve. Fourteen of the 86 patients (16.3%) in whom adrenal morphology was studied revealed adrenal gland enlargement on abdominal CT scan. ACTH stimulation was done in 12 of these 14 patients and eight of them had compromised adrenal reserve. Repeat ACTH stimulation done six months to one year after treatment in 13 patients revealed significantly increased 30 minutes (p < 0.05) and 60 minutes (p < 0.05) serum cortisol values. While nine of these 13 patients were negative responders before treatment, only three of them had evidence of compromised adrenal reserve after one year of antituberculosis treatment, (p < 0.05). Serum cortisol values in patients with drug-sensitive and drug-resistant tuberculosis did not differ significantly. Patients with drug-resistant tuberculosis had a higher prevalence of adrenal gland enlargement (7 of the 30) as compared to those with drug-sensitive tuberculosis (7 of the 56) (p = NS). Subclinical adrenal insufficiency is prevalent in a significant number of patients with both drug-sensitive and drug-resistant tuberculosis, and in some of these it is associated with adrenal gland enlargement. The compromised adrenal reserve and enlargement seem to reverse with therapy.

Effect of long-term administration of melatonin on adrenal histomorphology of the Indian desert gerbil Meriones hurrianae Jerdon.

Melatonin treatment for 8 weeks resulted in a significant increase (P < 0.01) in weights of the adrenal glands, in both male and female, M. hurrianae. The weight increase was not related to the time of administration of melatonin. Histology of adrenal glands in melatonin treated gerbils revealed a stimulatory response. In treated gerbils, adrenal cortex exhibited significant hypertrophy with enlarged nuclei and degranulated cytoplasm. Signs of hypertrophy were also evident in the adrenal medulla. The physiological significance of this response is not yet clear. It remains to be clarified if the hypertrophy indicates enhanced synthesis of corticosteroids.

Histopathological changes in aluminium phosphide poisoning.

A total of thirty cases of aluminium phosphide poisoning were studied for gross and microscopic changes in various organs of the body ie, lungs, liver, kidneys, heart, brain, stomach and adrenals. The histopathological changes revealed varying degrees of congestion, oedema and leucocytic infiltration, changes suggestive of cellular hypoxia. The most dramatic effects were produced in lungs, kidneys and adrenals.

Estudio bioquímico-histológico de los efectos de litio en algunos organos de vizcacha (Lagostomus maximus maximus) / Biochemical-histological study of the effects of lithium in some organs of vizcacha (Lagostomus maximus maximus)

Considerando que la vizcacha (lagostomus maximus maximus) es un roedor muy sensible al Litio (Li), planificamos un estudio bioquímico-histológico inyectando cloruro de Li 1 mEq/Kg/día vía intraperitoneal durante un mes a vizcachas adultas de ambos sexos agrupadas en tres lotes. En el lote I se comprobó por técnicas de microscopía óptica un evidente dano renal, gonodal, hipofisario y adrenal; por métodos bioquímicos se comprobó que el Li disminuyó significativamente los niveles séricos de LH en hembra, el contenido de testosterona y estradiol no fue modificado. En machos el Li sérico resultó significativamente mayor (p<0.01, Prueba de t) que en hembras. El dano fue mayor en machos. en el Lote II se estudió a 30 días de la última administración el grado de recuperación tisular del dano comprobado en el Lote I. Se verificó una recuperación total en hipófisis, parcial en testículo, nula en rinón y se incrementó el dano en adrenal y ovario. Con respecto al efecto seletivo sobre la zona glomerulosa adrenal del Li sería producido via hipófisis. En conclusión el efecto selectivo de Li en adrenal y gónadas en este roedor es una contribución para alertar sobre la posible producción de estos efectos en humanos

Drug induced endocrine problems.

Drug induced effects on the various endocrine and metabolic processes constitute a very common differential diagnostic possibility in investigating endocrine disorders. Knowledge of drug-effect is of paramount importance to avoid unnecessary and expensive investigations and treatment. Since most of the effects are type A effects and hence predictable, reduction of dose or modification of therapy is sufficient to reverse the adverse effects.

Effect of Staphylococcus protein-A on adrenal gland in mice.

Staphylococcus protein-A (SpA) was administered (ip) to Balb/c male mice for two weeks, twice a week at the dose level of 1, 6 and 12 micrograms in 200 microliters of normal saline. A significant change in the relative weights of liver, spleen, thymus, tracheobronchial lymph node, lung and testis was observed in 1 microgram SpA treatment group. In the adrenal, marked changes at the dose level of 1 microgram SpA treatment after 2 weeks were observed in the form of cellular proliferation on zona glomerulosa (ZG) and zona fasciculata (ZF) of adrenal cortex. With 6 and 12 micrograms SpA, adrenocortical masses were observed outside the capsule of adrenal gland. The enhanced effects of SpA at 4 weeks after treatment with the dose level of 6 and 12 micrograms were in the form of adrenal cell masses outside adrenal gland and histological changes in the adrenal cortex. The results suggest that long term and high doses therapy with SpA may be a risk factor to sensitive endocrine glands.

Mitomycin C induced inhibition of adrenal steroidogenesis in female rats.

Mitomycin C (MC), an antibiotic which depresses DNA synthesis causes suppression of enzyme delta 5 3 beta-hydroxysteroid dehydrogenase (delta 5 3 beta OHD) and glucose-6 phosphate dehydrogenase (G-6 PD) in the rat adrenal tissue. The treatment resulted in a fall in DNA content together with an accumulation of cholesterol and ascorbic acid in the gland. The results suggest a diminution in adrenal steroid biogenesis similar to gonadal inhibition previously reported.