Seasonal variations in the histology of the pituitary gland of Cirrhinus mrigala (Ham.) an Indian freshwater major carp, in relation to gonadal activity.

1. Seasonal variations in the histology of the pituitary gland of Cirrhinus mrigala (Ham.), an Indian freshwater major carp, in relation to gonadal activity has been studied during a one year period commencing, January, 1971. 2. The proximal pars distalis undergoes a great deal of seasonal variation in size, weight and nature of pituicytes. 3. The percentage composition of acidophils and cyanophils in the proximal pars distalis show a reciprocal relationship during the year. The acidophils predominate during the restitution phase of the gonads while the cyanophils preponderate among the different cell types during the breeding season. 4. The chromophobes do not show any significant fluctuation in numerical abundance in relation to gonadal activity. 5. Cyanophils of the proximal pars distalis undergo changes with change in gonadal activity Concentration of glycoproteinaceous material is extremely low during the restitution phase but high during the maturation phase. The cyanophils also undergo hypertrophy and hyperplasia during the spawning period. The latter discharge their contents and become vacuolated during the post spawning period. 6. The number and size of the globules in the cyanophils are related to gonadal activity. 7. Acidophils present in the rostral pars distalis and pars intermedia do not appear to be related to gonadal activity in C. mrigala. 8. The average weight of the gland is directly related to the state of maturation of the gonads, an observation being recorded here for the first time.     

Cyclic changes in the pituitary gland in correlation with the testicular cycle in a hill-stream teleost, Schizothorax richardsonii (Gray and Hard).

In Schizothorax richardsonii, the pituitary gland has been studied in correlation with the phases of the testicular cycle. The cyanophils of the proximal pars distalis show hypertrophy, hyperplasia, granulation, degranulation and vacuolisation in concurrence with the spermatogenic cycle. Secretory activity of the cyanophils controls the maturation of testes, which indicates that these cyanophils are gonadotrops. The formation of sperm mother cells, however, does not appear to depend on the secretory activity of the pituitary. During spawning and regression phases, the azocarminophilous granules in the cytoplasm of the cyanophils show quantitative variation. The migratory chromophil cells show numerical variation, suggesting their involvement in the reproductive cycle. Neurosecretory material also appears to play some part in reproduction.     

Pituitary cytology of the teleost fish Tilapia mossambica (Peters)

In the pituitary of Tilapia mossambica, eight cell types were identified on the basis of their staining reactions. the RPD consists of erythrosinophils and PbH positive cells. PbH cells border the NH and give amphipilic reaction to tri- and tetrachrome dyes and Halmi's stain. Erythrosinophils are also stained with acid fuchsin and orange G. The PPD is made up of acidophils and cyanophils. The acidophils are stained with orange G, acid fuchsin and erythrosin. These cells form a palisade zone between the neurohypophysis and the cyanophils. The cyanophils are AB, AF, ATh, PAS and aniline blue positive. They include both thyrotrophs and gonadotrophs and cannot be differentiated. The gonadotrophs may be those which are degranulated and vacuolated in the breeding season. The PI is formed of PAS- and PbH positive cells which lack any definite pattern of arrangement. Apart from the chromophils, scattered chromophobes were seen throughout the adenohypophysis. Occasionally, cells resembling the cyanophils of PPD were noticed in the RPD and PI also.     

Histophysiology of the pituitary gland in correlation with the ovarian cycle in Tor (Barbus) tor (Ham.)

Summary The histophysiology of the pituitary gland has been studied in correlation with the phases of the ovarian cycle in Tor (Barbus) tor. The cyanophilic cells in the proximal pars distalis display changes related to stages in the maturation of the ovary. The cyanophils, which display hypertrophy and hyperplasia and undergo degranulation and vacuolation during maturation, prespawning, and spawning periods, are identified as gonadotrops. Apparently their glycoproteinaceous contents, gonadotropin(s), control the processes of vitellogenesis, ovarian maturation, and initiation of oviposition. Depletion of glycoproteinaceous contents in these cyanophils leads to atresia of oocytes at advanced stages, but it does not appear to interfere with oogonial proliferation. The neurosecretory materials are implicated in female reproduction.I am deeply indebted to Professor H. Swarup for suggesting and supervising this study and to Dr. Grace E. Pickford for her generosity in sending me xerox copies of some articles.     

Pituitary cytology of the freshwater teleost Anabas testudineus (Bloch)

In the pituitary of A. testudineus the rostral pars distalis (RPD), proximal pars distalis (PPD) and pars intermedia (PI) are arranged in a rostro-caudal axis. The rostral neurohypophysis (NH) extends dorsal to the pars distalis, while the caudal part vertically penetrates into the PI and ramifies in its component. The RPD mainly consists of erythrosinophils and PgH-positive cells which are comparable to the lactotropes and corticotropes mentioned in the literature. In the nonbreeding season, acidophils are the predominant cells of the PPD and are largely confined to its dorsal aspect. The two types of cyanophils present in the PPD could not be differentiated with the various techniques used in this study. However, those cyanophils which increase in number and are active during the spawning season may be the gonadotropes, while those which are cytologically inactive may be the thyrotropes. The pars intermedia consists of PAS+ and PhH+ cells. Trichrome and tetrachrome staining techniques revealed the presence of acidophils, cyanophils and amphiphils in the PI.     

Changes in the Pituitary-interrenal Axis after Gonadectomy in the Catfish,Clarias batrachus (Linn.)

The effects of gonadectomy on the cytological structure of the hypophysis and interrenal tissue of Clarias batrachus have been investigated. Forty days after gonadectomy remarkable hypertrophy of certain cyanophils, identified later as gonadotrops, has been observed followed by hyperplasia. The somatotrops also seem to undergo similar changes, and as a result the proximal pars distalis (PPD) increases enormously and encroaches upon the pars intermedia (PI). The gonadotrops undergo a gradual degranulation and become devoid of stainable cytoplasm by 90 days. No signet ring cells have been noticed at any stage. The cyanophils in the ventral region of the PPD, identified as thyrotrops, also show degranulation, but to a comparatively limited degree. The ACTH cells undergo gradual hypertrophy and degranulation. A progressive increase is noticed in the nuclear diameter. The interrenal cells are activated as indicated by the enhanced nuclear indices. It is suggested that a fall in gonadal steroids may result in ACTH release which in turn activates the interrenal cells.     

Concentrations of somatotropin and prolactin in the follicular fluid and blood serum of cows in different phases of the estrous cycle

Comparative investigations of somatotropin and prolactin contents in the fluid of antral follicles and blood serum of cows in different phases of the oestrous cycle were performed. The somatotropin concentration in the fluid was shown to rise with increasing the follicle diameters from 3-5 to 6-10 mm in the follicular phase and to decrease in follicles of diameter 11-20 mm in the luteal phase. The prolactin concentration was higher in the fluid of follicles 11-20 mm in diameter than in those of 3-5 mm in diameter in the follicular phase and did not depend on the follicle size in the luteal phase. Concurrently, the prolactin content in follicles 3-5 mm in diameter was higher in the luteal than follicular phase of the cycle. As compared to the follicular phase, an increase in the prolactin concentration in the bovine blood serum during the luteal phase was also found. The data obtained indicate that changes in the somatotropin and prolactin contents in the follicular fluid are related to processes regulating growth and development of antral follicles depending on the phase of oestrous cycle and to changes in the blood hormone concentrations as well.     

Use of adenosine 5'-triphosphate as an indicator of the microbiota biomass in rumen contents

A number of techniques were tested for their efficiency in extracting adenosine 5'-triphosphate (ATP) from strained rumen fluid (SRF). Extraction with 0.6 N H(2)SO(4), using a modification of the procedure described by Lee et al. (1971), was the most efficient and was better suited for extracting particulate samples. Neutralized extracts could not be stored frozen before assaying for ATP because large losses were incurred. The inclusion of internal standards was necessary to correct for incomplete recovery of ATP. The ATP concentration in rumen contents from a cow receiving a ration of dried roughage (mainly alfalfa hay) ranged from 31 to 56 mug of ATP per g of contents. Approximately 75% of the ATP was associated with the particulate material. The ATP was primarily of microbial origin, since only traces of ATP were present in the feed and none was found in "cell-free" rumen fluid. Fractionation of the bacterial and protozoal populations in SRF resulted in the isolation of an enriched protozoal fraction with a 10-fold higher ATP concentration than that of the separated rumen bacteria. The ATP pool sizes of nine functionally important rumen bacteria during the exponential phase of growth ranged from 1.1 to 17.6 mug of ATP per mg of dry weight. This information indicates that using ATP as a measure of microbial biomass in rumen contents must be done with caution because of possible variations in the efficiency of extraction of ATP from rumen contents and differences in the concentration of ATP in rumen microbes.     

N-acetyltransferase activity and indole contents of the male Syrian hamster Harderian gland: changes during the light:dark cycle

The activities of N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT) and the indole contents of the Harderian glands of male Syrian hamsters were studied throughout a 24-h period. NAT activity exhibited a sharp rise 1 h after lights on, decreasing to basal levels 1 h later. Neither a HIOMT activity nor a melatonin concentration rhythm was detected throughout the 24 h. The 5-hydroxytryptamine (serotonin) concentration was highest during the dark phase reaching a peak at 0300 h; with light onset serotonin levels exhibited a rapid short-term drop. The 5-hydroxytryptophol concentration was highest during the mid- to late photophase; the lowest values to this constituent were measured late in the dark phase and at 1 h after lights on. The 5-hydroxyindole acetic acid concentration of the Harderian glands was rather stable throughout the 24-h period but levels did show a short-lived drop 1 h after light onset. Only a few animals contained detectable amounts of N-acetyl-5-hydroxytryptamine (N-acetylserotonin) in their Harderian glands. In agreement with previous work on the Harderian glands of female Syrian hamsters, the present results in males suggest that light onset is associated with marked changes in Harderian indoleamine metabolism.     

Influence of cumene hydroperoxide and 4-hydroxynonenal on the glutathione metabolism during in vitro ageing of human skin fibroblasts

Cumene hydroperoxide (Chp) and 4-hydroxynonenal (HNE) were used to investigate the effect of peroxidative challenge upon the glutathione (GSH) metabolism of human skin fibroblasts. Cellular GSH contents decreased during short-term incubations with Chp and oxidised glutathione (GSSG) was formed concomitantly. During longer incubations the GSH level was restored and the substrate flux through the pentose phosphate shunt increased. So in the presence of hydroperoxides the GSH level is maintained by reduction of GSSG. HNE caused a strong decrease in cellular GSH contents. Prolonged incubation with HNE lead to a rise in GSH contents above the basal level. The flux through the pentose phosphate shunt did not change during exposure to HNE. Hence, during incubation with HNE the cell maintains its GSH content by de novo synthesis of GSH. This conclusion is further substantiated by the findings with a cell strain deficient in GSH synthetase. These cells survived if incubated with Chp but not if exposed to HNE. GSH contents of normal cells from phase II (young) cultures and from phase III (aged) cultures responded similarly to Chp during short-term incubations and during a week of culture with the test compound. The flux through the pentose phosphate shunt rose much more in phase III than in phase II cells when incubated with the same concentration series of Chp. We conclude that during in vitro ageing the amount of NADPH needed to maintain cellular GSH levels in the presence of hydroperoxides increases, while the capacity to respond to such a challenge is not affected.     

Allocation of the suppressive activity of normal peripheral blood lymphocytes induced by diffusion chamber culture and Con A stimulation to the G2 phase of the cell cycle

Peripheral blood mononuclear cells stimulated either by diffusion chamber culture or by a high Con A concentration exhibit suppressive activity under conditions where no increase in cell number takes place. Instead an accumulation of large cells is observed which, according to their DNA contents, are classified as cells in the G2 phase of the cell cycle. By elutriation separation the suppressive activity is shown to be confined to this cell cycle phase.     

Regulation of the synthesis of 3-hydroxy-3-methylglutaryl coenzyme A reductase in the bovine ovary in vivo and in vitro

In order to investigate the pattern of ovarian cholesterol biosynthesis during the bovine estrous cycle, tissue concentrations of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, a rate-limiting enzyme in the synthesis of cholesterol, were determined by immunoblot techniques. Medium-sized (9-11 mm) and large (14-18 mm) follicles, after removal of follicular fluid by centrifugation, and corpora lutea from the early, early-mid, late-mid, and late stages of the luteal phase were used (n = 5 per group). The specific content (per microgram of tissue homogenate protein) and total content of HMG-CoA reductase in medium-sized and large follicles were substantially lower than those of corpora lutea of the early-mid and late-mid luteal phase. The specific content was elevated in a number of the corpora lutea from the early luteal phase and was low in regressing corpora lutea. Thus during the midluteal phase, when steroid hormone production is elevated, the total and specific contents of HMG-CoA reductase are also elevated. To investigate the mechanisms whereby the levels of HMG-CoA reductase are regulated, primary monolayer cultures of bovine luteal cells (early-mid and late-mid luteal phase) were used. Cells were cultured for 24 h in Dulbecco's modified Eagle's medium containing lipoprotein-poor fetal calf serum (2% vol/vol). At this concentration there was no stimulation of the production of progesterone above that seen with no addition of serum. Under these conditions the total and specific contents, and the synthesis, of HMG-CoA reductase were stimulated by treatment with (Bu)2cAMP (1 mM).(ABSTRACT TRUNCATED AT 250 WORDS)     

Use of Adenosine 5′-Triphosphate as an Indicator of the Microbiota Biomass in Rumen Contents

A number of techniques were tested for their efficiency in extracting adenosine 5′-triphosphate (ATP) from strained rumen fluid (SRF). Extraction with 0.6 N H₂SO₄, using a modification of the procedure described by Lee et al. (1971), was the most efficient and was better suited for extracting particulate samples. Neutralized extracts could not be stored frozen before assaying for ATP because large losses were incurred. The inclusion of internal standards was necessary to correct for incomplete recovery of ATP. The ATP concentration in rumen contents from a cow receiving a ration of dried roughage (mainly alfalfa hay) ranged from 31 to 56 μg of ATP per g of contents. Approximately 75% of the ATP was associated with the particulate material. The ATP was primarily of microbial origin, since only traces of ATP were present in the feed and none was found in “cell-free” rumen fluid. Fractionation of the bacterial and protozoal populations in SRF resulted in the isolation of an enriched protozoal fraction with a 10-fold higher ATP concentration than that of the separated rumen bacteria. The ATP pool sizes of nine functionally important rumen bacteria during the exponential phase of growth ranged from 1.1 to 17.6 μg of ATP per mg of dry weight. This information indicates that using ATP as a measure of microbial biomass in rumen contents must be done with caution because of possible variations in the efficiency of extraction of ATP from rumen contents and differences in the concentration of ATP in rumen microbes.     

Mechanism for the cholesterol-lowering action of egg white protein in rats

Eggs are a popular source of dietary cholesterol, but their consumption does not necessarily result in an increased serum cholesterol concentration. We investigated the cholesterol-lowering activity of egg white protein (EWP) and its potential mechanism in rats. The consumption of EWP resulted in a decreased concentration of cholesterol in the serum, liver and intestinal mucosa. The excretion of fecal neutral sterols and bile acids was greater by rats fed with EWP than by those fed with casein. The ratio of cholesterol and bile acids in the micellar phase to those in the solid phase was lower in the intestinal contents from rats fed with EWP than from those fed with casein. These results suggest that the cholesterol-lowering activity of EWP can be attributed to lowering the cholesterol absorption by intervening in the micellar formation in the intestines.     

Seasonal study of the adrenal gland of some Indian avian species.

Adrenal glands of eight Indian species of birds, namely Columba livia, Passer domesticus, Corvus splendens, Acridotheres tristis, Acridotheres ginginianus, Milvus migrans, Francolinus pondicerianus and Bubulcus ibis were examined during the sexually active and inactive phases of their annual reproductive cycles. Excepting A ginginianus and M. migrans, among members of either sex of the remaining six species the weight of the adrenal gland increases during the period of sexual activity. Histologically, the interrenal tissue of these birds could be divided into a peripheral subcapsular zone and a central zone. The cytochemical content of these two zones varies between sexual activity and inactivity. In sexually active birds of both sexes, interrenal cells of the central zone exhibit an increased concentration of alkaline phosphatase, glycogen, acid mucopolysaccharides and gross lipids, while in the subcapsular interrenal cells there is a prominent increase of ascorbic acid content. Cytochemical contents of chromaffin cells remain unchanged except acid phosphatase, which increases during the sexually active phase.     

The hypophysis of the marine teleost fish, Mugil auratus Risso.

Owing to its lack of a hypophysial stalk, the hypophysis of the marine fish, Mugil auratus, is of the platybasic type. Its differentiation into adenohypophysial and neurohypophysial regions well conforms with the situation already recognized in other fishes. But, although the adenohypophysis shows distinction into pro-, meso- and meta-adenophypophysis, the lineo-vertical arrangement which they assume is not of common occurrence. By contrast, the neurohypophysis forms the usual central nervous core, surrounded by the adenohypophysial regions. The acidophils constituting the pro-adenohypophysis, having a deep staining affinity, resemble those in the meso-adenohypophysial region. The acidophils of the meso-adenohypophysis occupy a small region, whereas their associating cyanophils occupy a larger ventral region. The meta-adenohypophysis consists of an integral convoluted sac of epithelial amphiphilic cells. Such encloses the bulk of the neurohypophysis. It is remarkable that the neurohypophysial fibres never interdigitate among the meta-adenohypophysial cells, so they are more closely related in arrangement to tetrapods than to fishes.     

Architectural remodeling of the tonoplast during fluid-phase endocytosis

During fluid phase endocytosis (FPE) in plant storage cells, the vacuole receives a considerable amount of membrane and fluid contents. If allowed to accumulate over a period of time, the enlarging tonoplast and increase in fluids would invariably disrupt the structural equilibrium of the mature cells. Therefore, a membrane retrieval process must exist that will guarantee membrane homeostasis in light of tonoplast expansion by membrane addition during FPE. We examined the morphological changes to the vacuolar structure during endocytosis in red beet hypocotyl tissue using scanning laser confocal microscopy and immunohistochemistry. The heavily pigmented storage vacuole allowed us to visualize all architectural transformations during treatment. When red beet tissue was incubated in 200 mM sucrose, a portion of the sucrose accumulated entered the cell by means of FPE. The accumulation process was accompanied by the development of vacuole-derived vesicles which transiently counterbalanced the addition of surplus endocytic membrane during rapid rates of endocytosis. Topographic fluorescent confocal micrographs showed an ensuing reduction in the size of the vacuole-derived vesicles and further suggest their reincorporation into the vacuole to maintain vacuolar unity and solute concentration.     

Zinc and copper in human milk

Studies of zinc and copper contents of breast milk were performed in 34 mothers during their lactation period until 5 days postpartum. They experienced no problems during pregnancy and delivery. All women had full term healthy infants, 22 were multiparous and 12 primiparous. The highest zinc concentration was found in the colostrum, and subsequently the concentration declined. Copper content was stable during the 5 days of lactation. Significantly lower zinc concentration and higher copper concentration were found in the serum of lactating women 3 days post partum, when compared with non-lactating women. Factors including lactation history, age of lactating women, serum levels of zinc and copper did not affect the contents of these trace minerals in breast milk.     

Flow cytometric analysis of unbalanced control of protein accumulation and DNA synthesis in differentiating mouse myeloid leukemia cells

The protein and DNA contents of mouse myeloid leukemia M1 (clone B24) cells were determined by flow cytometry (FCM) after double fluorescent staining of the cells with fluorescein isothiocyanate and propidium iodide. FCM analysis showed that there was a linear relationship between the DNA and protein contents in logarithmically growing cells, although the protein content showed some variation. B24 cells can be induced to differentiate into macrophage-like cells by treatment with a protein inducer(s) in conditioned medium (CM) of hamster embryo cells. When the cells were treated with various concentrations of CM, cells with a 2C DNA content, G1/0 cells, increased and protein accumulated in these G1/0 cells. The increases in the number of G1/0 cells and in their protein content per cell were proportional to the concentration of CM. Serial analysis of changes in the contents of DNA and protein in differentiating B24 cells showed that DNA synthesis was suppressed by differentiation-induced block of the cell cycle at the G1/0 phase, whereas increase in the protein content was not completely suppressed by block of the cell cycle. These results suggest that unbalanced control of the DNA and protein contents of B24 cells is involved in the mechanisms of the morphological changes during differentiation into macrophages.     

Processing of vegetable-borne carotenoids in the human stomach and duodenum

Carotenoids are thought to diminish the incidence of certain degenerative diseases, but the mechanisms involved in their intestinal absorption are poorly understood. Our aim was to obtain basic data on the fate of carotenoids in the human stomach and duodenum. Ten healthy men were intragastrically fed three liquid test meals differing only in the vegetable added 3 wk apart and in a random order. They contained 40 g sunflower oil and mashed vegetables as the sole source of carotenoids. Tomato purée provided 10 mg lycopene as the main carotenoid, chopped spinach (10 mg lutein), and carrot purée (10 mg beta-carotene). Samples of stomach and duodenal contents and blood samples were collected at regular time intervals after meal intake. all-trans and cis carotenoids were assayed in stomach and duodenal contents, in the fat and aqueous phases of those contents, and in chylomicrons. The cis-trans beta-carotene and lycopene ratios did not significantly vary in the stomach during digestion. Carotenoids were recovered in the fat phase present in the stomach during digestion. The proportion of all-trans carotenoids found in the micellar phase of the duodenum was as follows (means +/- SE): lutein (5.6 +/- 0.4%), beta-carotene (4.7 +/- 0.3%), lycopene (2.0 +/- 0.2%). The proportion of 13-cis beta-carotene in the micellar phase was significantly higher (14.8 +/- 1.6%) than that of the all-trans isomer (4.7 +/- 0.3%). There was no significant variation in chylomicron lycopene after the tomato meal, whereas there was significant increase in chylomicron beta-carotene and lutein after the carrot and the spinach meals, respectively. There is no significant cis-trans isomerization of beta-carotene and lycopene in the human stomach. The stomach initiates the transfer of carotenoids from the vegetable matrix to the fat phase of the meal. Lycopene is less efficiently transferred to micelles than beta-carotene and lutein. The very small transfer of carotenoids from their vegetable matrices to micelles explains the poor bioavailability of these phytomicroconstituents.