Effect on histological and sperm kinetics in DBP exposed Wistar rats.

Di-n-butyl phthalate (DBP) is a ubiquitous environmental pollutant, extensively used as a softener for polyvinyl chloride resins. A study was conducted to evaluate its effect on reproductive function of Wistar rats. DBP was given orally at a dose of 500, 1000 and 1500 mg kg(-1) body weight for 7 days. Evaluating histological and fertility parameters assessed reproductive function. Significant reduction in seminiferous tubule diameter, Leydig cell nuclear diameter (except at dose 500 mg), number of primary spermatocytes, secondary spermatocytes and spermatids were observed. Caudal sperm density and viability reduced significantly. Decrease in serum testosterone was also observed. Evidence indicates that DBP exposure causes dose dependent testicular toxicity and has the potential to induce adverse effect.

Testicular protein profile (SDS-PAGE) study of zinc deficient Wistar albino rat.

Present study has revealed that zinc plays important role in regulating the production and secretion of proteins at transcriptional or translational level. Study has firmly depicted the change in the levels of polypeptide of 70 kDa in zinc deficient group. The protein pattern in pair fed group has been affected mainly to combat the insult due to low food intake.

Short-term zinc deficiency in diet induces increased oxidative stress in testes and epididymis of rats.

In order to determine the effects of Zinc deficient diet on oxidative stress in testis and epididymis, various parameters viz: total proteins, lipid peroxidation, hydroperoxides, antioxidant capacity and enzymatic activities are evaluated in rats fed on zinc deficient diet for 2, 4 and 6 weeks. Total proteins, water and lipid solouble antioxidant capacity decreased while lipid peroxidation (TBARS) and hydroperoxides concentration increased in testes, caput and cauda epididymis except in 2ZD (testes) where hydroperoxides revealed a significant decrease. GSH decreased in testes and caput and cauda epididymis. GPx and gamma-GT activities increased in testes and caput and cauda epididymis of zinc deficient rats. Further, GST increased in testes but exhibited decreases after 2 and 4 weeks and an increase after 6 weeks in caput and cauda epididymis of zinc deficient rats. GR activities decreased in testes but it increased in caput and cauda epididymis of zinc deficient rats. Thus, zinc deprivation results in increased sensitivity to oxidative stress. All these may have been as a consequence of increased ROS generation and/or decreased zinc dependent antioxidant processes.

Metallothionein in male reproductive organs of adrenalectomized and hydrocortisone-treated Wistar rats.

Adrenalectomy resulted in an increase in metallothionein (MT) levels in testes, caput and cauda epididymis and prostate of rats but not in seminal vesicles where its levels decreased significantly. Inspite of administration of hydrocortisone, MT in testes, prostate (1.2 mg), caput (0.3 mg days 2, 8; 0.6 mg and 1.2 mg) and seminal vesicles (0.3 mg day 2, 4; 0.6 mg and 1.2 mg) remained increased. Thus adrenal insufficiency/hydrocortisone has no direct influence on MT levels. However, the increased levels of MT can be related to its ability to protect the cells from free radical damage caused by atrophy of reproductive tissues in adrenalectomised rats. Exogenously administered hydrocortisone to ADX rats resulted in return to ADX state as hydrocortisone metabolizes (half-life < 12 hr) and hence MT levels remained increased. The observations could provide a clue for the physiological functioning of the male reproductive tissue in a state of adrenal deprivation and hormonal supplementation.

Effect of dietary zinc deficiency on metallothionein concentration of epididymal luminal fluids of weanling Wistar albino rats.

Metallothionein (NIT) and zinc concentrations have been estimated in luminal fluids of caput/corpus and cauda epididymis and serum of zinc deficient (ZD), pairfed (PF) and control--ad libitum fed (ZC) groups of Wistar rats. MT decreased significantly in luminal fluids of caput corpus and cauda epididymis and serum of zinc deficient rats as compared to their respective controls. However, the decrease was non-significant in luminal fluids of corpus epididymis and serum of 4-weeks zinc deficient animals as compared to their control. Zinc levels also declined significantly in luminal fluids of epididymis and serum of zinc deficient rats as compared to their respective pairfed and control groups. Thus zinc deficiency state reduces zinc and MT concentrations in luminal fluid of epididymis and serum.

SDS-PAGE analysis of caput epididymis proteins in rats receiving a zinc deficient diet.

Caput epididymis proteins from control, pairfed and zinc deficient (ZD) wistar weanling albino rats after 2-, 4-, 6- and 8-weeks were examined using SDS-PAGE followed by densitometric scanning of the gels. In comparison to the control and pairfed rats, ZD rats displayed new proteins. These included a Mr 42 kDa from 2ZD, Mr 47.5, 27.5, 23.2 and 16.0 kDa from 4ZD and Mr 87 and 14.2 kDa from 6ZD group. The 8ZD group, however, revealed no additional protein bands over controls. Further, several other proteins were missing from ZD rats. These included Mr 93 and 71 kDa from 2ZD; 93, 90, 79, 67, 62, 55 and 15.3 kDa from 4ZD; 60, 45.5, 34, 30 and 24 kDa from 6ZD and 41.5, 33 and 27.5 kDa bands from 8ZD group. The results indicate that the induced Zn-deficient state may be responsible for the altered protein patterns in the caput epididymis. The duration of low Zn uptake period also appears to influence the protein pattern in caput epididymis.

Zinc, copper and hydrolytic enzymes in epididymis of hydrocortisone treated rat.

Administration of glucocorticoid (1, 2 and 4 mg) in excess leads to degeneration of epididymides as supported by cellular degeneration, sperm density and morphometric measurements. Zinc level increased statistically after 1, 2 and 4 mg hydrocortisone treatment while copper increased after 1 and 2 mg treatment. Cholesterol, protein and leucine aminopeptidase levels increased and decreased significantly in caput and cauda respectively. Activity of alkaline phosphatase reduced significantly while the treatment of hydrocortisone at different doses elevated acid phosphatase, aryl sulphatase and lactate dehydrogenase activities. Evidently, these changes are as a result of onset of cellular degeneration leading to impairment of metabolic/secretory activity of epididymal cells. The possible involvement of pituitary-testis axis in hydrocortisone induced epididymal degeneration and functional inhibition has been discussed.

Effect of adrenalectomy and adrenalectomy+hydrocortisone treatment on histopathological, biochemical and zinc and copper profiles in rat testes.

Degenerative changes such as decreased seminiferous tubule diameter, Leydig cell nuclear diameter, spermatogenic arrest, oedematous fluid in the interstitium and lumen of seminiferous tubules and increased levels of zinc, copper and enzymes (lactate dehydrogenase, LDH; leucine aminopeptidase, LAP; and aryl sulphatase) in adrenalectomised rats suggest a possible role of adrenal cortex and its hormones in spermatogonial cell proliferation and subsequent differentiation, homeostasis of biological trace elements and behaviour of enzymes. Atrophy of Leydig cells and the degenerative changes in testes of adrenalectomised rats can be attributed to reduced supply of testosterone. Hydrocortisone, administered through a single dose acted as hyperstate of hydrocortisone for a short duration, thereby inhibiting steroidogenesis either directly by affecting Leydig cell testosterone production or indirectly by affecting the release of LH from pituitary gland and thus caused degeneration of germinal epithelium. Once hydrocortisone (half life < 12 hr) was metabolized, the animals returned to adrenalectomised state, the degeneration persisted. Thus, hydrocortisone administered through a single dose was insufficient to sustain spermatogenesis. Chronic administration at physiological dose may renew spermatogenesis. Increased levels of LDH, LAP and arylsulphatase are, probably, necessary for cellular degeneration. Zinc and copper exhibited an increase and the rise can be corroborated to (1) failure of regulatory mechanism(s) that control the flow of the elements across the blood-testes barrier; and (2) increased oedematous fluid formed by cellular deaths of the germinal epithelium.

Light and electron microscopic changes in the ovary of zinc deficient BALB/c mice.

Female BALB/c strain of mice fed on Zn deficient diet for 2-, 4- and 6- weeks exhibited prolonged diestrous phase with only VII types of follicles instead of VIII as compared to their respective control and pairfed. Light microscopic studies displayed increased atresia, cessation of oogenesis and ovulation, degeneration of follicular cells of zona granulosa, clumped chromatin of oocyte and disrupted zona pellucida and corona radiata. Ultrastructural studies of peripheral follicular and theca interstitial cells of type VI and VII follicles revealed swollen mitochondria, dilated ERs (free of RNP particles), increased lysosomes, several necrotic areas of cytoplasm and pyknotic nuclei. Conclusively, Zn deficiency may lead to (1) reduction in energy, protein intake and in secretion of GnRH by hypothalamus and LH and FSH by hypophysis, (2) increased synthesis and/or secretion of prolactin. (3) reduced output of estrogen, and (4) eventually slow growth or arrest of ovulation or atresia of the growing follicles in the ovary.

Histological and biochemical changes in testis of zinc deficient BALB/c strain of mice.

Zinc, protein, cholesterol, phospholipids, alkaline phosphatase (AlPase), acid phosphatase (AcPase), adenosine-5-triphosphatase(ATPase) and histology were studied in testis of zinc-deficient mice. Zinc and protein decreased in the 3-week experiment whereas they increased in the 6-week experiment. Zinc is involved in several functions of the cell and is regulated by hormones. Inhibition of spermatogenesis indicates for decreased zinc levels in 3-week whereas the increase in 6-week experiment indicates for accumulation of zinc in oedomatous fluid and uncontrolled diffusion of zinc across the blood testis barrier. Glycogen decreased in the 3-week as well as 6-week experiments due to blockage of androgen and spermatogenesis. Cholesterol and phospholipids increased in the 3-week experiment and decreased in 6-week experiment as both the parameters are related to steroidogenesis. Zinc deficiency leads to aspermatogenic condition and comparatively less injury to non-germinal cells. This could have blocked the transport of material across the testis barrier and therefore might have increased AlPase levels. Increased AcPase, probably represents lysosomal enzymes, as the cell debris of disorganised epithelium are to be digested and removed. ATPase increased in 3-week experiment and can be correlated to increased demands of energy of testicular cells to overcome the insults of zinc deficiency whereas the decrease in 6-week experiment could be as a result of inhibition of spermatogenesis.

Ultrastructural study of epidermal Langherhans cells in leprosy.

ATPase staining and ultrastructural study of skin biopsies from six patients of leprosy (2TT, 4LL) and three normal subjects was carried out to study Langherhans Cells (LC). ATPase staining showed normal counts of LCs in tuberculoid patients, while significant reduction was observed in lepromatous cases. Electron microscopy revealed morphological changes in LL cases in the form of dense matrix and indistinct cristae of mitochondria; decreased number of lysosomes and rough endoplasmic reticulum; and numerous vacuoles in cytoplasm. TT cases showed normal morphology. Possible role of Langerhans Cells in pathogenesis of leprosy is discussed.