Growth and Protein Pattern in Cowpea Seedlings Subjected to Salinity

Cowpea (Vigna unguiculata) seeds were put to germinate on filter paper under control (distilled water) and salt stress (100 mM MaCl) conditions. Seeds and seedlings were classified in eight developmental stages (DS), according to their morphological traits. Under control conditions, 7 d after planting, 100 % of the seedlings reached DS VIII (seedlings with radicles measuring more than 5 cm, cotyledons leaving the filter paper, hypocotyls straight and cotyledonary leaves fully open) and under NaCl stress conditions, 11 d after planting only 68 % of the seedlings were at DS VIII. The length of the main root and of shoot has decreased 23 and 44 %, respectively. The two-dimensional electrophoretic patterns of the albumins isolated from stems and leaves were determined in seedlings at DS VIII. In stems 19 proteins (14.6 to 76.3 kDa) had their relative concentration increased by salinity, 8 (31.2 to 65.0 kDa) had their relative concentration decreased by salinity and 9 (16.3 to 39.8 kDa) were apparently synthesised de novo. In leaves, under salt conditions 9 proteins (18.2 to 33.2 kDa) increased in concentration, one (17.1 kDa) decreased in concentration and one (21.2 kDa) was apparently synthesised de novo. This revised version was published online in July 2006 with corrections to the Cover Date.     

Estradiol influences the LH response to met-enkephalin. An immunocytochemical and morphometric study.

In order to test the possible role of estradiol in the response of the LH-adenohypophyseal cells to the administration of met-enkephalin in the albino male rat, an immunocytochemical (peroxidase-antiperoxidase), morphometric (cellular and nuclear areas and numerical density) and radioimmunoassay (LH serum levels at the moment of sacrifice) study was carried out. The intraventricular administration of met-enkephalin (150 micrograms in 25 microliters of distilled water) does not produce any changes in the parameters considered. However, when the animals were pretreated with estradiol (chronically, 15 days), met-enkephalin produced a significant decrease in all the parameters considered.     

Vitamin D-Binding Protein in Healthy Pre- and Postmenopausal Women: Relationship with Estradiol Concentrations

Objective: To examine the relationship between endogenous serum estradiol and vitamin D–binding protein (DBP) and total, free, and bioavailable 25-hydroxyvitamin D (25OHD) concentrations in pre- and postmenopausal women.Methods: In 165 healthy women (ages, 26 to 75 years) not taking any form of exogenous estrogen, the serum concentrations of estradiol, 25OHD, DBP, parathyroid hormone, and albumin were measured. Free and bioavailable 25OHD (free + albumin-bound) levels were calculated from total 25OHD, DBP, and serum albumin levels.Results: Premenopausal women had higher serum 25OHD (31.5 ± 7.9 ng/mL), DBP (45.3 ± 6.2 mg/dL), and estradiol (52.8 ± 35.0 pg/mL) levels than postmenopausal women (26.5 ± 4.9 ng/mL, 41.7 ± 5.7 mg/dL, and 12.9 ± 4.9 pg/mL), respectively. In addition, the calculated free and bioavailable 25OHD levels were higher in prethan postmenopausal women (P<.05). Serum estradiol correlated with DBP (r = 0.22; P<.01) and total 25OHD (r = 0.27; P<.01). In multivariate regression models (with or without serum 25OHD), estradiol was independently associated with DBP (P<.05).Conclusion: Lower estradiol level is one of the factors that contribute to lower DBP levels in older women. Our data indicate that besides well-known factors such as age, gender, and race, serum estradiol concentrations are also a physiologic predictor of DBP concentration.Abbreviations: 25OHD = 25-hydroxyvitamin D BMI = body mass index CV = coefficient of variation DBP = vitamin D–binding protein PTH = parathyroid hormone SHBG = sex hormone–binding globulin     

Luteinizing Hormone (LH) Response to Different Doses of Synthetic Gonadotropin Releasing Hormone (Gnrh) in Prepubertal Gilts

The object of this investigation was to study luteinizing hormone (LH) response to different doses of synthetic gonadotropin-releasing hormone (GnRH) in prepubertal gilts. Four crossbred prepubertal gilts, 128–134 days old and body weight 57–63 kg, were used in this study. Four doses, 0. 5, 25 and 125 μg, of GnRH were administered via a jugular vein catheter in a latin square design. Each treatment consisted of 3 injections at 90 min intervals. Frequent blood samples were taken during a period of 90 min before up to 90 min after treatment. Total LH responses were measured from post-treatment samples as the area under the curve above base level obtained from pre-treatment samples. A positive relationship between GnRH dose and LH release was obtained in all gilts, except for 1 treatment given to a gilt with high plasma level of oestradiol-17β on the day of treatment. This study has demonstrated the responsiveness of the pituitary gland by LH release to different doses of GnRH in 4.5-month-old prepubertal gilts.     

LH release in mink (Mustela vison). Pattern of the LH surge and effect of metabolic status

The mink is a seasonal breeder with induced ovulation and delayed implantation. Reproductive processes are strongly influenced by energy supply and body condition. Items for which there is paucity or complete lack of data were the main objectives of this study: the temporal relationship between copulation and the pre-ovulatory LH surge and the influence of energy supply on LH release. A total of 30 yearling female mink with a well defined metabolic status was used. Twelve females kept in the laboratory were measured in six consecutive one-week balance periods each including the measurement of heat production by means of indirect calorimetry, and 18 females were kept under conventional farm conditions. The animals were fed so as to maintain energy balance (CON), flush fed by 2 weeks food restriction followed by 2 weeks refeeding (FLUSH), or kept in a negative energy balance (NEG). Plasma concentrations of the thyroid hormones, IGF-1 and insulin were determined weekly (n = 12), or 1 week after change in energy supply to the FLUSH group (n = 18). On the day of mating, blood samples for LH and oestradiol-17beta (E2) were taken before and immediately after mating and then 4, 8, 12, 24, 30 and 48 h thereafter. Frequent blood samplings, each lasting 60 min, were taken during the LH surge from two other females surgically fitted with venous access ports. Peak concentrations of LH were recorded on the first sampling, an average 16 min after mating. The concentrations remained elevated for 12 h, but almost decreased to basal values 24 h after mating. Plasma E2 was high before mating and peak values were attained 4 h after mating after which it decreased. Energy supply had no significant influence on LH and E2, but there was a tendency for a more sluggish LH release in NEG animals. The lack of response in FLUSH animals was explained by these animals having a lower intake of metabolisable energy than CON animals, the total intake not being significantly different from the NEG group. Plasma concentrations of thyroid hormones, IGF-1 and insulin were not significantly affected by the treatment, but in FLUSH animals the values mirrored energy supply, and in the NEG group, the values tended to decrease during the course of the experiment. It was concluded that the pre-ovulatory LH surge is an immediate response to mating, and that reproductive activity in the mink is maintained over a wide range of energy supply and body condition.     

Spatial and Temporal Pattern of Caddisfly Distribution at a Mesohabitat Scale in two Patagonian Mountain Streams Subjected to Pastoral Use

Substrate, flow type, nutrients, aquatic vegetation, organic matter, and caddisfly community structure were studied at two low order streams (Glyn and Nant y Fall) subjected to pastoral use in the Patagonian mountains. At both sites, we examined the effect of habitat type (boulder‐pebble with and without filamentous algae, cobble‐pebble, gravel‐sand, leaf‐pack, the submerged macrophytes Myriophyllum quitense and Isoetes savatieri) and season (high and low water period) on caddisfly assemblages. Benthic particulate organic matter (BPOM) ranged between 4.6 and 472 g m^–2, all allochtonous detrital fractions were significantly higher at leaf‐packs at Glyn, whereas M. quitense habitats supported more BPOM and macrophytes biomass at Nant y Fall. As expected, boulder‐pebble sustained higher Trichoptera richness than M. quitense and gravel‐sand, moreover all habitats showed higher density than M. quitense at Nant y Fall. According to our results at least nine caddisfly species exhibited some habitat preference with boulder‐pebble and cobble‐pebble the most selected habitat. These particular habitats sustained more than 68% of the total caddisfly species. Multidimensional scaling ordination highlighted differences in composition per habitat for both sites showing a clear distinction among depositional and erosional habitats. Substrate, flow type, detritus biomass were important predictors defining assemblages. Based upon our findings, those anthropogenic actions or stressors that change hydraulic as well substrate attributes in mountain streams such as stock trampling, dredging, clearing of riparian areas, will reduce caddisfly richness. These results are relevant for outline management and conservation biomonitoring and schemes in headwater Patagonian streams as well as other similar environments worldwide (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Structure-Related Oxidative Damage in Rat Brain After Acute and Chronic Electroshock

The role of oxidative stress in electroconvulsive therapy-related effects is not well studied. The purpose of this study was to determine oxidative stress parameters in several brain structures after a single electroconvulsive seizure or multiple electroconvulsive seizures. Rats were given either a single electroconvulsive shock or a series of eight electroconvulsive shocks. Brain regions were isolated, and levels of oxidative stress in the brain tissue (cortex, hippocampus, striatum and cerebellum) were measured. We demonstrated a decrease in lipid peroxidation and protein carbonyls in the hippocampus, cerebellum, and striatum several times after a single electroconvulsive shock or multiple electroconvulsive shocks. In contrast, lipid peroxidation increases both after a single electroconvulsive shock or multiple electroconvulsive shocks in cortex. In conclusion, we demonstrate an increase in oxidative damage in cortex, in contrast to a reduction of oxidative damage in hippocampus, striatum, and cerebellum.     

The Orientation and Dynamics of Estradiol and Estradiol Oleate in Lipid Membranes and HDL Disc Models

Estradiol (E₂) and E₂ oleate associate with high-density lipoproteins (HDLs). Their orientation in HDLs is unknown. We studied the orientation of E₂ and E₂ oleate in membranes and reconstituted HDLs, finding that E₂ and E₂ oleate are membrane-associated and highly mobile. Our combination of NMR measurements, molecular dynamics simulation, and analytic theory identifies three major conformations where the long axis of E₂ assumes a parallel, perpendicular, or antiparallel orientation relative to the membrane’s z-direction. The perpendicular orientation is preferred, and furthermore, in this orientation, E₂ strongly favors a particular roll angle, facing the membrane with carbons 6, 7, 15, and 16, whereas carbons 1, 2, 11, and 12 point toward the aqueous phase. In contrast, the long axis of E₂ oleate is almost exclusively oriented at an angle of ∼60° to the z-direction. In such an orientation, the oleoyl chain is firmly inserted into the membrane. Thus, both E₂ and E₂ oleate have a preference for interface localization in the membrane. These orientations were also found in HDL discs, suggesting that only lipid-E₂ interactions determine the localization of the molecule. The structural mapping of E₂ and E₂ oleate may provide a design platform for specific E₂-HDL-targeted pharmacological therapies.     

Ultrastructure of rat pituitary LH gonadotrophs in relation to serum and pituitary LH levels following repeated LH-RH stimulation

The effects of single and repeated LH-RH injections at 120 min intervals on female rat LH gonadotrophs and on pituitary and serum LH levels were investigated using electronmicroscopy and radioimmunoassay. A temporary stimulation of granule release, of protein and new granule synthesis and of the accumulation of lysosomal structures was found in LH cells after the first LH-RH injection. The temporary stimulations were massively enhanced after the second injection. These consecutive yet in their time-sequence overlapping processes account for the initial depletion of secretory granule content (3--15 min after LH-RH injection), for the subsequent regranulation and accumulation of granules above control levels (60--120 min after injection) and also for the reduction in the number of granules to control levels (150 min after LH-RH injection and thereafter). Increased polymorphic lysosomal structures are believed to be responsible for this reduction of excess granules. The amount of assayable pituitary and serum LH generally corresponds with the morphological changes observed in LH-gonadotrophs, thus further substantiating the above observations. A schema which summarizes the observed morphological and hormonal changes in their time-sequence in response to LH-RH stimulation depicts the short-term regulation of secretory processes in female gonadotrophs.     

Clinical,Morphological and Endocrinological Studies in Gilts with Delayed Puberty

Thirty-six gilts which had not shown oestrus at about 8 months of age or more were transported from the pig research station to the clinic, a journey of 12 km. The gilts were examined by laparoscopy and those which had only small follicles in the ovaries were catheterized and placed in pens, with sexually mature boars kept in adjacent pens. Oestrus detection was done twice daily and blood was sampled three times a day. After 7 days the laparoscopy was repeated and gilts which still had only small follicles in their ovaries were given 250 μg GnRH intravenously the following day. Blood samples were taken frequently before and after GnRH treatment. One week. after administration of GnRH the ovaries were inspected by laparoscopy once more. The first laparoscopic examination showed that 42 % of the gilts were sexually mature. One gilt had no uterus or ovaries. Twenty gilts had only small follicles in the ovaries and fourteen of these gilts showed ovulatory oestrus 5.5 days (4-7.5 days) after arrival. In these fourteen gilts a rise in the oestradiol-17B level (>30 pmol/1) was seen at an average time of 1.9 days and a rise in LH (preovulatory peak) was seen at an averaged 4.5 days after the start of blood sampling. Six gilts were given 250 ug GnRH. An immediate rise in LH could be seen in all the gilts (mean peak level was 6.18 μg/l) and the elevated levels had a duration of 4 hours. None of the GnRH-treated gilts responded with oestrus symptoms or increased ovarian activity.     

Controlling interrelationships of progesterone/LH and estradiol/LH in mares

The interrelationships of progesterone, estradiol, and LH were studied in mares (n=9), beginning at the first ovulation (Day 0) of an interovulatory interval. An increase in mean progesterone concentrations began on Day 0 and reached maximum on Day 6, with luteolysis beginning on Day 14. A common progesterone threshold concentration of about 2 ng/ml for a negative effect on LH occurred at the beginning and end of the luteal phase. Progesterone and LH concentrations decreased at a similar rate from Day 6 until the onset of luteolysis on Day 14, consistent with a decreasing positive effect of LH on progesterone. Concentrations of LH during the increase in the ovulatory surge consisted of two linear regression segments involving a rate of 0.4 ng/ml/day for Days 14-22 and 1.8 ng/ml/day for Day 22 to 1 day after the second ovulation. The end of the first segment and beginning of the second segment was 2 days before ovulation and was the day the ovulatory estradiol surge was at a peak.     

Polyribosome Content in Young and Aged Wheat Leaves Subjected to Drought

The effect of a droughting treatment on the polyribosome contentof wheat leaves was markedly different on leaves of differentages. Growing leaves showed a large decrease in their polyribosomepopulation, while fully expanded leaves showed no loss of polyribosomescompared with controls. These results suggest that the largereductions in polyribosome population observed in growing tissuesare due to reductions in growth, rather than a direct effectof water stress.     

Rapid Absorption and Local Redistribution of Progesterone after Vaginal Application in Gilts

Catheters were implanted in 6 anaesthetized gilts (3 animals in the follicular phase, 3 in the luteal phase) into a carotid artery and into the utero-ovarian vein and uterine artery on both sides. The uterine lumina were closed by a suture. Further, a catheter was inserted into the vagina after which the animals were allowed to recover. Tritiated progesterone was infused into vagina the following day during a 2 min period and simultaneous blood samples collected from the 5 catheters every 10 min for 2 h after which the animals were sacrificed. Tissue samples were obtained from the genital organs. The results showed a rapid absorption of progesterone from the vaginal lumen and a marked redistribution to the genital organs. The increased level of radioactivity in the plasma samples collected from the uterine arteries compared to the simultaneous samples from the carotid artery in 2 of the 3 animals in the luteal phase indicates the existence of a local redistribution system.