Effect of diethylstilbesterol and prolactin on the induction of follicle stimulating hormone receptors in immature and cycling rats

Induction of follicle stimulating hormone receptor in the granulosa cells of intact immature rat ovary by diethylstilbesterol, an estrogen, has been studied. A single injection of 4 mg of diethylstilbesterol produced 72 h later a 3-fold increase in follicle stimulating hormone receptor concentration as monitored by [125I]-oFSH binding to isolated cells. The newly induced receptors were kinetically indistinguishable from the preexisting ones, as determined by Lineweaver-Burk plot of the binding data. The induced receptors were functional as evidenced by increased ability of the granulosa cells to incorporate [3H]-leucine into cellular proteins. Neutralization of endogenous follicle stimulating hormone and luteinizing hormone by administering specific antisera had no effect on the ability of diethylstilbesterol to induce follicle stimulating hormone receptors, whereas blockade of endogenous prolactin secretion by ergobromocryptin administration significantly inhibited (∼ 30 %) the response to diethylstilbesterol; this inhibition could be completely relieved by ovine prolactin treatment. However, ovine prolactin at the dose tried did not by itself enhance follicle stimulating hormone receptor level. Administration of ergobromocryptin to adult cycling rats at noon of proestrus brought about as measured on diestrusII, (a) a reduction of both follicle stimulating hormone (∼ 30 %) and luteinizing hormone (∼ 45 %) receptor concentration in granulosa cells, (b) a drastic reduction in the ovarian tissue estradiol with no change in tissue progesterone and (c) reduction in the ability of isolated granulosa cells to convert testosterone to estradiol in response to follicle stimulating hormone. Ergobromocryptin treatment affected only prolactin and not follicle stimulating hormone or luteinizing hormone surges on the proestrus evening. Treatment of rats with ergobromocryptin at proestrus noon followed by an injection of ovine prolactin (1 mg) at 1700 h of the same day completely reversed the ergobromocryptin induced reduction in ovarian tissue estradiol as well as the aromatase activity of the granulosa cells on diestrus II, thus suggesting a role for proestrus prolactin surge in the follicular maturation process.

Standardization and validation of an induced ovulation model system in buffalo cows: Characterization of gene expression changes in the periovulatory follicle

In bovines characterization of biochemical and molecular determinants of the dominant follicle before and during different time intervals after gonadotrophin surge requires precise identification of the dominant follicle from a follicular wave. The objectives of the present study were to standardize an experimental model in buffalo cows for accurately identifying the dominant follicle of the first wave of follicular growth and characterize changes in follicular fluid hormone concentrations as well as expression patterns of various genes associated with the process of ovulation. From the day of estrus (day 0), animals were subjected to blood sampling and ultrasonography for monitoring circulating progesterone levels and follicular growth. On day 7 of the cycle, animals were administered a PGF2α analogue (Tiaprost Trometamol, 750 μg i.m.) followed by an injection of hCG (2000 IU i.m.) 36 h later. Circulating progesterone levels progressively increased from day 1 of the cycle to 2.26 ± 0.17 ng/ml on day 7 of the cycle, but declined significantly after PGF2α injection. A progressive increase in the size of the dominant follicle was observed by ultrasonography. The follicular fluid estradiol and progesterone concentrations in the dominant follicle were 600 ± 16.7 and 38 ± 7.6 ng/ml, respectively, before hCG injection and the concentration of estradiol decreased to 125.8 ± 25.26 ng/ml, but concentration of progesterone increased to 195 ± 24.6 ng/ml, 24 h post-hCG injection. Inh-α and Cyp19A1 expressions in granulosa cells were maximal in the dominant follicle and declined in response to hCG treatment. Progesterone receptor, oxytocin and cycloxygenase-2 expressions in granulosa cells, regarded as markers of ovulation, were maximal at 24 h post-hCG. The expressions of genes belonging to the super family of proteases were also examined; Cathepsin L expression decreased, while ADAMTS 3 and 5 expressions increased 24 h post-hCG treatment. The results of the current study indicate that sequential treatments of PGF2α and hCG during early estrous cycle in the buffalo cow leads to follicular growth that culminates in ovulation. The model system reported in the present study would be valuable for examining temporo-spatial changes in the periovulatory follicle immediately before and after the onset of gonadotrophin surge.

Effect of LHRH agonists and antagonists in male and female bonnet monkeys (Macaca Radiata)

A few analogues of LHRH have been tested in the adult bonnet monkeys using change in serum testosterone following LHRH injection as a parameter of response to LHRH. Of the four analogues tested in male monkeys, Buserelin was found to be the most potent one in increasing serum testosterone levels. Injection of the LHRH antagonist at 1600 h resulted in the abolition of the characteristic nocturnal surge of testosterone observed in adult bonnet monkeys maintained under regulated light conditions. Following administration of LHRH a/s during early pregnancy, serum chorionic gonadotropin levels decreased though the course of pregnancy was not affected. These results suggest that bonnet monkey can be successfully employed to test LHRH analogues.

Regulation of Testosterone Rhythmicity by Gonadotropins in Bonnet Monkeys (Macaca radiata)

In an attempt to study the factor(s) that regulates production of nychthemeral testosterone surges in adult male bonnet monkeys (Macaca radiata), serum levels of testosterone, LH, FSH, and prolactin were monitored during a 24 h period. Only prolactin showed a significant increment in its levels coincident with that of the testosterone surge. The relationship between LH and testosterone production was studied by 1) observing the responsiveness of testes, in terms of testosterone production, to one or two injections of oLH (1 mg/injection) given 12 h apart at 0900 and 2100 h; and 2) monitoring the effect on testicular testosterone production of LH antiserum injection given at 1000, 1700, and 2100 h. That each LH injection brought about an increment in testosterone level of equal magnitude suggests that the difference in responsiveness of the testes to unchanging levels of LH at morning and night hours is not due to any alteration in substrate availability at the two time intervals. The LH antiserum experiments indicate that irrespective of the time of its administration the nocturnal surge of testosterone, which normally occurs at 2200 h, is blocked. While the antiserum prevents a rise in testosterone level, it appears not to influence basal testosterone production. The results further show that even at 2100 h, when surge testosterone production is already initiated, the testis is still highly sensitive to lack of LH, antiserum injection bringing about within 2 h a significant reduction in testosterone levels (by 69% in experimentals vs 11% in controls).

Dynamics of Circulating Concentrations of Gonadotropins and Ovarian Hormones Throughout the Menstrual Cycle in the Bonnet Monkey: Role of Inhibin A in the Regulation of Follicle-Stimulating Hormone Secretion

In higher primates, increased circulating follicle-stimulating hormone (FSH) levels seen during late menstrual cycle and during menstruation has been suggested to be necessary for initiation of follicular growth, recruitment of follicles and eventually culminating in ovulation of a single follicle. With a view to establish the dynamics of circulating FSH secretion with that of inhibin A (INH A) and progesterone (P-4)secretions during the menstrual cycle, blood was collected daily from bonnet monkeys beginning day 1 of the menstrual cycle up to 35 days. Serum INH A levels were low during early follicular phase, increased significantly coinciding with the mid cycle luteinizing hormone (LH) surge to reach maximal levels during the mid luteal phase before declining at the late luteal phase, essentially paralleling the pattern Of P-4 secretion seen throughout the luteal phase. Circulating FSH levels were low during early and mid luteal phases, but progressively increased during the late luteal phase and remained high for few days after the onset of menses. In another experiment, lutectomy performed during the mid luteal phase resulted in significant decrease in INH A concentration within 2 hr (58.3 +/- 2 vs. 27.3 +/- 3 pg/mL), and a 2- to 3-fold rise in circulating FSH levels by 24 hr (0.20 +/- 0.02 vs. 0.53 +/- 0.14 ng/mL) that remained high until 48 hr postlutectomy. Systemic administration of Cetrorelix (150 mu g/kg body weight), a gonadotropin releasing hormone receptor antagonist, at mid luteal phase in monkeys led to suppression of serum INH A and P-4 concentrations 24 hr post treatment, but circulating FSH levels did not change. Administration of exogenous LH, but not FSH, significantly increased INH A concentration. The results taken together suggest a tight coupling between LH and INH A secretion and that INH A is largely responsible for maintenance of low FSH concentration seen during the luteal phase. Am. J. Primatol. 71:817-824, 2009.

Proportional Weirs as Velocity Controlling Devices

Of the many factors that govern the settling phenomenon, the flow velocity in the settling tanks can be controlled favorably by fixing suitably designed weirs at the outlets of the tanks. The velocity at the bottom should not dislodge the particles that have already settled. These requirements might be met with by velocities which are controlled to be constant with respect to the depth of flow, or velocities which reduce linearly with increasing depth or velocities that vary inversely with the depth. To achieve these types of velocity control, new proportional weirs have been designed. Very near to the outlet of the tank, over a small length, the flow was found to be turbulent and noncompliant with the expected type of velocity control. This small length of the disturbance may be provided over and above the theoretical settling length of the tank, for efficient sedimentation.

Anode Phenomena in Triggered Vacuum Gaps

Arc voltage - current characteristics and threshold current densities occuring during the formation of anode spots in triggered vacuum gaps are reported. Single pulses of 1.65 ms arcing time, which correspond to switching surge currents, are used in the study with copper and aluminum anodes. The threshold values are 1.75 times the values reported earlier using the longer, 8 mis, arcing time. They are found to depend upon the duration of arcing time as well as upon electrode material, surface conditions, electrode size and contact separation. Lateral inhomogenity in the electrode geometry appears to reduce the threshold value by promoting early formation of anode spots.

Shape effect on optimal geometric conditions in surface aeration systems

The performance of surface aeration systems, among other key design variables, depends upon the geometric parameters of the aeration tank. Efficient performance and scale up or scale down of the experimental results of an aeration ystem requires optimal geometric conditions. Optimal conditions refer to the conditions of maximum oxygen transfer rate, which assists in scaling up or down the system for ommercial utilization. The present work investigates the effect of an aeration tank's shape (unbaffled circular, baffled circular and unbaffled square) on oxygen transfer. Present results demonstrate that there is no effect of shape on the optimal geometric conditions for rotor position and rotor dimensions. This experimentation shows that circular tanks (baffled or unbaffled) do not have optimal geometric conditions for liquid transfer, whereas the square cross-section tank shows a unique geometric shape to optimize oxygen transfer.

Scale-up criteria of square tank surface aerator

Oxygen transfer rate and the corresponding power requirement to operate the rotor are vital for design and scale-up of surface aerators. Present study develops simulation or scale-up criterion correlating the oxygen transsimulation fer coefficient and power number along with a parameter governing theoretical power per unit volume (X, which is defined as equal to (FR1/3)-R-4/3, where F and R are impellers' Fronde and Reynolds number, respectively). Based on such scale-up criteria, design considerations are developed to save energy requirements while designing square tank surface aerators. It has been demonstrated that energy can be saved substantially if the aeration tanks are run at relatively higher input powers. It is also demonstrated that smaller sized tanks are more energy conservative and economical when compared to big sized tanks, while aerating the same volume of water, and at the same time by maintaining a constant input power in all the tanks irrespective of their size. An example illustrating how energy can be reduced while designing different sized aerators is given. The results presented have a wide application in biotechnology and bioengineering areas with a particular emphasis on the design of appropriate surface aeration systems.

Scale-up criteria of square tank surface aerator

Oxygen transfer rate and the corresponding power requirement to operate the rotor are vital for design and scale-up of surface aerators. Present study develops simulation or scale-up criterion correlating the oxygen transsimulation fer coefficient and power number along with a parameter governing theoretical power per unit volume (X, which is defined as equal to (FR1/3)-R-4/3, where F and R are impellers' Fronde and Reynolds number, respectively). Based on such scale-up criteria, design considerations are developed to save energy requirements while designing square tank surface aerators. It has been demonstrated that energy can be saved substantially if the aeration tanks are run at relatively higher input powers. It is also demonstrated that smaller sized tanks are more energy conservative and economical when compared to big sized tanks, while aerating the same volume of water, and at the same time by maintaining a constant input power in all the tanks irrespective of their size. An example illustrating how energy can be reduced while designing different sized aerators is given. The results presented have a wide application in biotechnology and bioengineering areas with a particular emphasis on the design of appropriate surface aeration systems.

Flow Generated By Pitched Blade Turbines Ii: Simulation Using Κ-Ε Model

Experimental data on average velocity and turbulence intensity generated by pitched blade downflow turbines (PTD) were presented in Part I of this paper. Part II presents the results of the simulation of flow generated by PTD The standard κ-ε model along with the boundary conditions developed in the Part 1 have been employed to predict the flow generated by PTD in cylindrical baffled vessel. This part describes the new software FIAT (Flow In Agitated Tanks) for the prediction of three dimensional flow in stirred tanks. The basis of this software has been described adequately. The influence of grid size, impeller boundary conditions and values of model parameters on the predicted flow have been analysed. The model predictions successfully reproduce the three dimensionality and the other essential characteristics of the flow. The model can be used to improve the overall understanding about the relative distribution of turbulence by PTD in the agitated tank

Analysis of Long Cantilever Cylindrical Shell Subjected to Wind Loading

Bending analysis of closed cylindrical shells subjected to asymmetric load and having different support conditions is of interest in the design of chimneys, water towers, oil storage tanks, etc. A simple method of analyzing a long cantilever cylindrical shell, subjected to asymmetric load, is presented in the paper, using Schorer’s shell theory and orthogonal functions. The application of the solution has been illustrated with an example of a cantilever shell subjected to wind loads. The results obtained for this problem have been compared with the previously available results to illustrate the accuracy of the results obtained here. The solution presented can also be extended to a cylindrical shell with other support conditions, as well as to the study of free vibration of a cylindrical shell. The present solution will be very useful for designers who need to obtain numerical results for specific problems with minimum computational effort.

Effect of follicle-stimulating hormone and its antiserum on the activity of ornithine decarboxylase in the ovary of rat and hamster

The ability of FSH to stimulate the activity of ornithine decarboxylase (ODC) in the ovary of the immature rat and cycling hamster has been examined using specific antisera to gonadotropins. The stimulatory effect of FSH on ODC activity in the ovary of the immature rat was abolished when LH antiserum was administered along with FSH, while similar administration of FSH antiserum had no effect on LH action in stimulating ODC activity, thereby demonstrating the specificity of the LH effect. During the estrus cycle of the hamster, ODC activity in the ovary could be detected only on the evening of proestrus, the maximal activity seen at 1700 h being associated with both the Graafian follicles and the rest of the ovarian tissue. Neutralization of the proestrous FSH surge had no effect on the activity of ODC in either of these tissues, while similar administration of LH antiserum at 1300 h of proestrus completely inhibited the ODC activity in both large follicles and the rest of the ovarian tissue. Thus, the surge of LH, but not of FSH, appears to be responsible for regulating the ODC activity in the ovary of the cycling hamster.

Use of adult male bonnet monkeys (Macaca radiata) for testing the biological activity of gonadotropin releasing hormone and its analogues

The effect of injecting agonistic and antagonistic analogues of gonadotropin releasing hormone analogues on serum testosterone levels was checked in adult and immature male bonnet monkeys. Of the agonistic analogues Buserelin, Ovurelin and D-Phe6 Gln8 GnRH were found to be most potent in increasing serum testosterone levels in the adult male bonnet monkeys. While 27-month-old monkeys responded well to des Gly10 GnRH, only marginal response was observed in the case of 15-month-old monkeys. Studies carried out with Ovurelin indicated that it was not effective in causing desensitization in adult monkeys. The antagonistic analogue was effective in blocking nocturnal surge of serum testosterone. Based on these studies it is suggested the adult male bonnet monkeys can be effectively used for testing the activity of GnRH analogues.

Multicode MIMO for High Data Rate Mobile Ad-hoc Networks

In a mobile ad-hoc network scenario, where communication nodes are mounted on moving platforms (like jeeps, trucks, tanks, etc.), use of V-BLAST requires that the number of receive antennas in a given node must be greater than or equal to the sum of the number of transmit antennas of all its neighbor nodes. This limits the achievable spatial multiplexing gain (data rate) for a given node. In such a scenario, we propose to achieve high data rates per node through multicode direct sequence spread spectrum techniques in conjunction with V-BLAST. In the considered multicode V-BLAST system, the receiver experiences code domain interference (CDI) in frequency selective fading, in addition to space domain interference (SDI) experienced in conventional V-BLAST systems. We propose two interference cancelling receivers that employ a linear parallel interference cancellation approach to handle the CDI, followed by conventional V-BLAST detector to handle the SDI, and then evaluate their bit error rates.