Adrenal testis inter-relationship in male Indian palm squirrel Funambulus pennanti Wroughton

The present work attempts to study adrenal-testes relationship in a seasonal breeder, Indian palm squirrel, Funambulus pennanti. To assess the functional status of the adrenals, its two major components i.e. cholesterol and ascorbic acid were estimated biochemically and the results related with testicular functioning and its cholesterol concentration. In addition, histological studies of the above tissues were also carried out. Variation in adrenal function is clearly manifested by fluctuations in its cholesterol and ascorbic acid contents. However, no alteration in adrenal histology was discernible. Testicular recrudescence starts in January and peak activity is reached in April and May when adrenals are most inactive. End of June markes the beginning of testicular regression and maximal regression is reached in October.     

Diurnal variations in circulating estradiol, testosterone, melatonin and harderian gland porphyrin concentration in Indian palm squirrel, Funambulus pennanti

A two-peak cyclicity in the plasma level of melatonin, estradiol/testosterone and Harderian porphyrin was noted in F. pennanti. An inverse relationship of Harderian porphyrin with plasma melatonin and a direct relation of it with plasma estradiol/testoserone level were also observed, suggesting that the variation of Harderian, porphyrin concentration may be under the control of both, circulating melatonin and gonadal steroids.     

Response of calcitonin cells, parathyroid glands and bone to prolonged calcitonin administration in the Indian palm squirrel, Funambulus pennanti (Wroughton)

The administration of calcitonin (4 MRC units injected intraperitoneally daily for 42 days) caused hypocalcaemia, hypophosphataemia and a decrease in the serum acid phosphatase level in adult Indian palm squirrels, Funambulus pennanti. Hypocalcaemia and the decrease in the serum acid phosphatase level persisted up to 21 days but the hypophosphataemic effect continued throughout the experiment. The serum calcium level increased from day 28 up to day 35 and the serum acid phosphatase level reached the control level at 28 days. At the close of the experiment (42 days), both serum calcium and acid phosphatase levels were again decreased. Calcitonin-induced hypocalcaemia resulted in hypertrophy of calcitonin cells which were densely packed with secretory granules up to day 21 of the treatment. Thereafter they displayed both hypertrophy and hyperplasia till the end of the experiment. Some calcitonin cells showed degranulation after 35 and 42 days of treatment. Few lead-haematoxylin-positive calcitonin cells with collapsed membranes and pyknotic nuclei were also seen in the lumina of the thyroid follicles toweards the close of the experiment. Parathyroid chief cells showed hypertrophy from 21 to 35 days of the treatment. From day 28 to the close of the experiment they released their secretory product. After 42 days of experimental treatment a growth-promoting effect of calcitonin was observed (increase in body weight and femur weight.     

Calcium, protease action, and the regulation of the cell cycle

New layered matrix systems designed for zero-order sustained release are presented. The devices consist of a hydrophobic middle layer and press-coated hydrophillic and/or hydrophobic barrier layer(s). The systems overcome the inherent disadvantage of non-linear release associated with diffusion-controlled matrix devices by providing additional releasing surface with time to compensate for the decreasing release rate. The in vitro release of pseudoephedrine hydrochloride as a model compound from the matrices was examined. Modulation of release from a selected matrix containing aminophylline was evaluated in a crossover study using nine beagle dogs. Preliminary in vitro/in vivo correlation was also studied. The systems described in this paper can potentially be scaled up to commercial production.     

Cell cycle regulation of the Saccharomyces cerevisiae polo-like kinase cdc5p

Progression through and completion of mitosis require the actions of the evolutionarily conserved Polo kinase. We have determined that the levels of Cdc5p, a Saccharomyces cerevisiae member of the Polo family of mitotic kinases, are cell cycle regulated. Cdc5p accumulates in the nuclei of G2/M-phase cells, and its levels decline dramatically as cells progress through anaphase and begin telophase. We report that Cdc5p levels are sensitive to mutations in key components of the anaphase-promoting complex (APC). We have determined that Cdc5p-associated kinase activity is restricted to G2/M and that this activity is posttranslationally regulated. These results further link the actions of the APC to the completion of mitosis and suggest possible roles for Cdc5p during progression through and completion of mitosis.     

Proliferation, apoptosis and cell cycle regulation in prostatic carcinogenesis

Besides the MutLS-like system, Schizosaccharomyces pombe has an additional pathway of mismatch repair. This minor pathway, producing short excision tracts, repairs C/C and, with lower efficiency, other mismatches also. We investigated the involvement of the exo1+, msh2+ and pms1+ genes in the two pathways. The exo1+ gene encodes a 5' to 3' exonuclease, while msh2+ and pms1+ are homologs of Escherichia coli mutS and mutL, respectively. Intragenic two-factor crosses showed that exo1+, msh2+ and pms1+ are involved in the major, but not in the C/C-correcting, pathway. Post-meiotic segregation frequencies and mitotic mutation rates in single and double mutants supported this finding. Furthermore, msh2 delta was epistatic over exo1 delta, and the ExoI enzyme is likely to be redundant with other exonucleases.     

Beta-adrenergic signal transduction in the hypothalamus of the European hamster: relation with the seasonal hibernation cycle and the diurnal activity cycle

In vivo nailfold capillary microscopy was performed on 10 men with vibration white finger (VWF) and 10 age-matched male controls. The observed nailfold capillaries required adaptation of Maricq's classifications and addition of new morphological scoring systems. These new classifications produced numerical scores for assessing capillary: dropout, tortuosity, elongation, visualization of subpapillary venular plexus, and the degree of disarrangement of nailfold capillary polarity. Application of these new scores showed, for the first time, a complex pattern of abnormal-nailfold capillaries in patients with VWF. Capillary dropout was evident in 7/10 patients, with an associated disarrangement in nailfold capillary polarity in five. All 10 controls had normal capillary morphology. Tortuosity of the capillary loops and elongation of their length was observed in 30% of patients. These significant morphological alterations seen in VWF suggest a local small-vessel vasculitis.     

Cell cycle regulation and differentiation in the human podocyte lineage

Mature podocytes are regarded as growth-arrested cells with characteristic phenotypic features that underlie their function. To determine the relationship between cell cycle regulation and differentiation, the spatiotemporal expression of cyclin A, cyclin B1, cyclin D1, the cyclin-dependent kinase inhibitors (CKIs) p27 and p57, and markers of differentiating podocytes in developing human kidneys was investigated by immunohistochemistry. In S-shaped body stage, Ki-67, a cell proliferation marker that labels the G1/S/G2/M phase, was expressed in the majority (more than 80%) of presumptive podocytes, along with cyclin A (approximately 20% of the Ki-67-positive cells) and cyclin B1 (less than 5% of Ki-67-positive cells) expression. Among these cells), cyclin D1 and CKIs were markedly down-regulated. At the capillary-loop stage, by contrast, CKIs and cyclin D1 were intensely positive in podocytes, whereas no Ki-67, cyclin B1, or cyclin A expression was seen. Moreover, double-immunolabeling and serial-section analysis provided evidence that CKIs and markers specific for differentiating podocytes, namely PHM-5 (podocalyxin-like protein in humans), synaptopodin (a foot process-related protein), and C3b receptor, were co-expressed at the capillary-loop stage. Podocytes were the only cells within the glomeruli that expressed CKIs at immunohistochemically detectable levels. Furthermore, bcl-2 (an apoptosis inhibitory protein) showed a reciprocal expression pattern to that of CKI. These results suggest that 1) the cell cycle of podocytes is regulated by cyclin and CKIs, 2) CKIs may act to arrest the cell cycle in podocytes at the capillary-loop stage, and 3) the specific cell cycle system in podocytes may be closely correlated with their terminal differentiation in humans.     

Cell cycle regulation of the double stranded RNA activated protein kinase, PKR

The interferon (IFN)-induced, double stranded RNA (dsRNA)-activated serine/threonine kinase, PKR, is a potent negative regulator of cell growth when overexpressed in yeast or mammalian cells. To determine whether endogenous PKR plays a role in cell growth control, we have investigated the regulation of PKR levels and activity during the cell cycle in human glioblastoma T98G cells. The steady-state level of PKR mRNA in T98G cells was highest in growth arrested cells, dropped sharply within 3 h of serum stimulation then gradually increased as cells progressed through G1, reaching a plateau in early S phase. PKR protein level increased following serum stimulation reaching a peak at the G2+M boundary and declining thereafter. In contrast, PKR kinase activity exhibited two peaks, in early G1 and at the G1/S boundary, declining sharply in early S phase. Thus, the activity profile did not follow the protein profile indicating a tight regulation of PKR at the level of activity. In T98G cells expressing the catalytically inactive PKRK296R the dsRNA-induced activation of NF-kappaB and IRF-1 was suppressed and the mutant cells exhibited resistance to stress induced apoptosis. Cell cycle distribution analysis showed that the mutant expressing cells exhibited longer G1 phase and fewer cells engaged in S phase. Furthermore, early passage mouse embryo fibroblasts derived from PKR knockout mice grew more slowly compared with the control cells. Taken together these results suggest that PKR may play a role in cell cycle progression.     

In vivo regulation of the early embryonic cell cycle in Xenopus

We report here the first extensive in vivo study of cell cycle regulation in the Xenopus embryo. Cyclin A1, B1, B2, and E1 levels, Cdc2 and Cdk2 kinase activity, and Cdc25C phosphorylation states were monitored during early Xenopus embryonic cell cycles. Cyclin B1 and B2 protein levels were high in the unfertilized egg, declined upon fertilization, and reaccumulated to the same level during the first cell cycle, a pattern repeated during each of the following 11 divisions. Cyclin A1 showed a similar pattern, except that its level was lower in the egg than in the cell cycles after fertilization. Cyclin B1/Cdc2 kinase activity oscillated, peaking before each cleavage, and Cdc25C alternated between a highly phosphorylated and a less phosphorylated form that correlated with high and low cyclin B1/Cdc2 kinase activity, respectively. Unlike the mitotic cyclins, the level of cyclin E1 did not oscillate during embryogenesis, although its associated Cdk2 kinase activity cycled twice for each oscillation of cyclin B1/Cdc2 activity, consistent with a role for cyclin E1 in both S-phase and mitosis. Although the length of the first embryonic cycle is regulated by both the level of cyclin B and the phosphorylation state of Cdc2, cyclin accumulation alone was rate-limiting for later cycles, since overexpression of a mitotic cyclin after the first cycle caused cell cycle acceleration. The activity of Cdc2 closely paralleled the accumulation of cyclin B2, but cell cycle acceleration caused by cyclin B overexpression was not associated with elevation of Cdc2 activity to higher than metaphase levels. Tyrosine phosphorylation of Cdc2, absent during cycles 2-12, reappeared at the midblastula transition coincident with the disappearance of cyclin E1. Cyclin A1 disappeared later, at the beginning of gastrulation. Our results suggest that the timing of the cell cycle in the Xenopus embryo evolves from regulation by accumulation of mitotic cyclins to mechanisms involving periodic G1 cyclin expression and inhibitory tyrosine phosphorylation of Cdc2.     

Effect of chronic naloxone and morphine treatments on testicular, body weight, and plumage pigmentation cycle of lal munia, Estrilda amandava

At Imphal (24 degrees 44' N) testes of lal munia, Estrilda amandava, began in June/July, peaked in September/ October, and thereafter declined to a minimum in December/January. Daily im treatments of 2.5-10 mg/kg/ bird/30 days of naloxone during progressive phase suppressed testicular growth, but without effects during quiescent, peak, and regression phases. Daily morphine (5 mg/kg/bird) during progressive and peak phases stimulated testicular growth, but without effects during quiescent and regression phases. Daily morphine (10 mg/kg/bird) during progressive phase stimulated the testes, an effect reversed by daily im treatments of an equivalent dose of naloxone. Seasonal changes in body weight closely correlated with testicular size. Daily im naloxone (5 and 10 mg/kg/bird/30 days) during progressive phase inhibited the increased body weight, but had no effects during quiescent, peak, and regression phases. Morphine (5 mg/kg/bird/day) during progressive and peak phases increased body weight, but had no effects during quiescent and regression phases. Morphine (10 mg/kg/bird/day) during progressive phase increased body weight, an effect which was reversed by equivalent dose of naloxone. Plumage color increased progressively between May and August/September, was maintained during October, and thereafter declined to reach dull-brown henny feathers by December. Daily im naloxone (2.5-10 mg/kg/bird/30 days) regardless of the reproductive states did not affect plumage color cycle. Morphine (5 mg/kg/ bird/day) accelerated plumage pigmentation between June and August, but had no effect during progressive or peak phases. Postnuptial decline in plumage color was inhibited by morphine (5 and 10 mg/kg/bird/day) and naloxone failed to reverse this effect. It is concluded that in the lal munia, endogenous opioid peptides are important constituents of the neuroendocrine mechanisms that influence development of the testes and body weight.     

A study of the nycthemeral cycle of behavioural temperature regulation in man

1. Four human subjects were rendered hyperthermic and hypothermic by immersion in warm and cool water, at 02.00, 08.00, 14.00 and 20.00 hr. Bath and oesophageal temperatures and pulse rate were recorded. Temperature preference was determined by operant behaviour and vote. The core temperature set-point for behavioural thermoregulation was estimated from the behavioural results. 2. The results are in accord with those of previous studies of the nyethemeral cycling of autonomic responsiveness to heat and cold with a heating up phase before noon and a cooling down phase during the early night. 3. Subjective sensations and behavioural responses were also found to follow a nycthemeral cycle with a minimum before noon and a maximum at 20.00 hr. 4. The core temperature set point was 0-7 degrees C higher after noon than before noon with a small phase advance from resting core temperature. This result suggests that the nycthemeral cyclic change in body temperature is due to a nycthemeral cyclic change in the set-point near to which body temperature is kept by both autonomic and behavioural thermoregulatory responses.     

Hydra hymanae: regulation of the life cycle by time and temperature

Hydra hymanae, a hermaphroditic freshwater coelenterate, reproduces asexually at 24 degrees C and sexually at 15 degrees C. The appearance of gonads begins 12 days after transfer from 24 degrees to 15 degrees C and is complete 35 days after the temperature transition. Testes appear before eggs. Fifty percent of the mature embryos maintained at 15 degrees C hatch by day 61, but they have a low level of survival. Fifty percent of the mature embryos pretreated for from 5 to 25 days at 4 degrees C hatch by about day 45, and these have a high level of survival. Embryos maintained at 4 degrees C for longer periods (55 to 85 days) accumulate in a prehatching state and hatch with a high degree of synchrony approximately 7.5 days after return to 15 degrees C. Populations derived from newly hatched polyps are refractory to sex induction for approximately 120 days. The system is well adapted to ensure a regular alternation of reproductive modes in the natural environment.     

The role of pheromones in the regulation of estrous cycle duration in the guinea pig

A decrease in estrous cycle duration, due to shortening of the period of vaginal closure (VC), has been observed in female guinea pigs exposed to the odor of urine from males of the same species. VC shortening was also observed in females exposed to the odor of female urine collected during the period of vaginal opening (VO). No alteration in VC duration occurred, however, in females exposed to urine collected in the 1st 7 days of VC. Also VC shortening did not occur in bulbectomized females exposed to the odor of male urine. Therefore, it was concluded that guinea pig urine, when highly concentrated, contains pheromones capable of shortening estrous cycle VC.     

Correlation between motility of testicular spermatozoa, testicular histology and the outcome of intracytoplasmic sperm injection

Many studies have demonstrated the postoperative analgesic efficacy of fentanyl delivered i.v. by patient-controlled analgesia (PCA) devices at demand doses ranging from 10 to 50 microg, but none has sought to define the optimal fentanyl PCA dose. In this randomized, double-blind, multicenter study, we compared the safety and efficacy of three administered demand-dose sizes of fentanyl (20, 40, and 60 microg) in 150 patients after major surgery. Efficacy was dose-dependent; positive response rates (i.e., a global assessment score of "very good" or "excellent" and the absence of severe opioid adverse effects) were 42%, 52%, and 68% for the 20, 40, and 60 microg demand-dose groups, respectively, and were significantly higher in the 60 microg demand-dose group. The number of doses administered and missed attempts were significantly smaller in the 40 and 60 microg demand-dose groups compared with the 20 microg demand-dose group. This suggests that the 20 microg demand dose provided inadequate pain relief. Adverse respiratory events were more frequent and mean respiratory rates were significantly slower with the 60 microg demand dose, compared with the 20 or 40 microg demand doses. These results indicate that, of these three doses, the 40 microg demand dose was optimal for fentanyl PCA management of moderate to severe pain after major surgery. IMPLICATIONS: The postoperative analgesic efficacy of fentanyl delivered i.v. by patient-controlled analgesia devices has been demonstrated for demand doses ranging from 10 to 50 microg, but the optimal fentanyl dose remains unknown. In this randomized, double-blind study, we compared three demand dose sizes of fentanyl (20, 40, and 60 microg) and found that the 40 microg demand dose was the most appropriate for fentanyl patient-controlled analgesia management of postoperative pain.     

Activities and androgenic regulation of kreb cycle enzymes in the epididymis and vas deferens of rhesus monkey

The activities of nine enzymes of the TCA cycle were estimated in the initial segment, caput, corpus and cauda segments of epididymis and vas deferens of adult rhesus monkey and expressed as units per mg DNA. These enzymes were also estimated in epididymal segments and vas deferens of castrated and castrated-androgen replaced monkeys as well. Results indicated higher activities of most of the enzymes in vas deferens as compared to epididymal segments. All the enzymes showed marked reduction in epididymis and vas deferens after castration, the effect being much more pronounced in the epididymis, than in the vas. Androgen replacement in castrated monkeys stimulated most of the enzymes markedly in epididymis and in the vas deferens as compared to their castrated values. The response of cauda and vas deferens to exogenous androgen treatment was however moderate, as compared to the other epididymal segments. The studies indicate that energy metabolism in the epididymis (as well as in the vas deferens) is strictly androgen dependent and the energy charge of these target organs is likely to fall appreciably after castration, which may in turn affect many energy dependent processes of these organs (e.g. absorption, secretion of specific substances etc.) which have been considered important for sperm maturation and survival.     

Altered regulation of cholesterol metabolism in type I diabetic women during the menstrual cycle

This study examines the relationship of cellular cholesterol metabolism to oestrogen and progesterone during the menstrual cycle in diabetic and non-diabetic subjects. Nine premenopausal diabetic women were compared to nine non-diabetic women of the same age. Oestrogen, progesterone, lipoproteins, including lipoprotein (a) (Lp(a)) and cholesteryl ester transfer protein (CETP) were determined in serum. Cellular cholesterol content and cellular cholesterol synthesis were measured in mononuclear leucocytes. There was no significant change in serum lipoproteins including Lp(a) during the cycle in either group. CETP activity was significantly higher over the 4 weeks in the diabetic patients compared with non-diabetic subjects (mean 463 +/- 30 mumol l-1 h-1 vs 405 +/- 28 mumol l-1 h-1, p < 0.01). Serum high density lipoprotein (HDL) cholesterol was significantly lower during the 4 weeks in the diabetic patients (1.7 +/- 0.1 mmol l-1 vs 1.8 +/- 0.1 mmol-1, p < 0.05). Cellular cholesterol synthesis decreased steadily up to the third week in cells from the control subjects whereas there was no significant change in cells from diabetic patients whose cellular cholesterol synthesis was higher at week 3 compared with non-diabetic subjects (663 +/- 54 nmol mg-1 cell protein vs 432 +/- 43 nmol mg-1 cell protein, two-way interaction p < 0.05). There was a significant negative correlation between cellular cholesterol synthesis and serum oestrogen in the non-diabetic subjects (p < 0.05) but not in the diabetic patients.(ABSTRACT TRUNCATED AT 250 WORDS)     

5-HT autoreceptors in the regulation of 5-HT release from guinea pig raphe nucleus and hypothalamus

5-HT autoreceptors involved in the regulation of 5-HT release in the guinea pig dorsal raphe nucleus have been studied in comparison with those in the hypothalamus. In vitro release was measured in slices of raphe and hypothalamus prelabelled with [3H]5-HT, superfused with Krebs solution and depolarized electrically. The non-selective 5-HT receptor agonist, 5-carboxamidotryptamine (5-CT) (0.1-10 nM for raphe: 1-100 nM for hypothalamus) and antagonist, methiothepin (10-1000nM), decreased and increased, respectively, the release of [3H]5-HT evoked by electrical stimulation in either of these regions when given alone. The selective 5-HT1B/D receptor antagonist, GR127935 (100-1000 nM), and the 5-HT1D receptor antagonist, ketanserin (300-1000 nM), had no significant effect on this release in either of these regions. Methiothepin and GR127935 (100-1000 nM) shifted to the right the concentration-effect curve of 5-CT in both the raphe and the hypothalamus. At 300 nM, ketanserin shifted to the right the concentration-effect curve of 5-CT in the raphe but did not modify the 5-CT curve in the hypothalamus. In microdialysis experiments ketanserin, applied locally at 10 microM, increased the extracellular levels of 5-HT in the dorsal raphe nucleus of the freely moving guinea pig, whereas 5-HT levels were unchanged in the hypothalamus. Ketanserin at 1 microM did not affect the decrease in 5-HT output induced by the selective 5-HT1B/D receptor agonist, naratriptan (used at 10 microM in raphe and 0.1 microM in hypothalamus), in the raphe or the hypothalamus. In the raphe, WAY100635, a 5-HT1A receptor antagonist, at 1 microM, did not prevent naratriptan (10 microM) from reducing the extracellular levels of 5-HT. These results suggest that, in the conditions used in this study, the release of 5-HT in the dorsal raphe nucleus is possibly modulated in part by 5-HT1B receptors but essentially the control is through 5-HT receptors whose subtype is still to be determined. In the hypothalamus, however, it is clear that only 5-HT1B receptors are involved in the modulation of 5-HT neurotransmission.