Testosterone inhibits gonadotropin-releasing hormone pulse frequency in the male sheep

The objective was to determine the effect of chronic testosterone (T) treatment on GnRH and LH secretion in wethers. Rams were either castrated only or castrated and immediately treated with Silastic implants containing T. Several weeks later, a device for collecting hypophyseal-portal blood was surgically implanted. Six to seven days later, blood samples were collected simultaneously and continuously from the portal vessels and jugular vein of pairs of conscious animals. Samples were divided at 10-min intervals for 6-12 h. One hour before the end of collection, all animals received i.v. injections of 250 ng of GnRH. In samples collected simultaneously from 6 pairs of animals, T reduced the frequency of both GnRH pulses (1.8 +/- 0.2 vs. 0.9 +/- 0.3/h, p less than 0.03) and LH pulses (1.6 +/- 0.1 vs. 0.8 +/- 0.3/h, p less than 0.03). T did not alter amplitude of either GnRH or LH pulses. Testosterone reduced mean GnRH (9.7 +/- 0.6 vs. 7.9 +/- 0.5 pg/ml, p less than 0.05), whereas mean LH was not significantly reduced (9.6 +/- 1.4 vs. 6.1 +/- 1.8 ng/ml, p = 0.16). These results support the hypothesis that T reduces GnRH pulse frequency.     

Loss of luteinizing hormone surges induced by chronic estradiol is associated with decreased activation of gonadotropin-releasing hormone neurons

Chronic exposure of young ovariectomized rats to elevated circulating estradiol causes loss of steroid-induced LH surges. Such LH surges are associated with cFos-induced activation of GnRH neurons; therefore, we hypothesized that chronic estradiol treatment abolishes LH surges by decreasing activation of GnRH neurons. Regularly cycling rats were ovariectomized and immediately received an estradiol implant or remained untreated. Three days or 2 or 4 wk later, the estradiol-treated rats received vehicle or progesterone at 1200 h, and 7 hourly blood samples were collected for RIA of LH. Thereafter, all rats were perfused, and the brains were examined for immunocytochemical localization of cFos and GnRH. The GnRH neurons from untreated ovariectomized rats rarely expressed cFos. As reported, LH surges induced by 3 days of estradiol treatment were associated with a 30% increase in cFos-containing GnRH neurons, and progesterone enhanced both the amplitude of LH surges and the proportion of cFos-immunopositive GnRH neurons. As hypothesized, the abolition of LH surges caused by 2 or more weeks of estradiol was paralleled by a reduction in the percentage of cFos-containing GnRH neurons, and this effect was delayed by progesterone. These results suggest that chronic estradiol abolishes steroid-induced LH surges in part by inactivating GnRH neurons.     

Pituitary desensitization and the regulation of pituitary gonadotropin-releasing hormone (GnRH) receptors following chronic administration of a superactive GnRH analog and testosterone

We recently demonstrated that chronic daily administration of a superactive GnRH analog to intact rats resulted in an initial stimulation of serum LH levels with a subsequent return of LH levels to baseline at a time when testosterone levels were marked decreased. These data demonstrated pituatary desensitization following chronic GnRH analog treatment. Administration of GnRH analog with a dose of testosterone which did not markedly lower serum LH levels when administered alone prevented the stimulation of LH secretion by analog. The present studies were undertaken to determine the effects of GnRH analog and testosterone administration on the regulation of pituitary GnRH receptors. Pituitary GnRH receptor binding was increased by analog treatment alone at 20 days and returned to control levels at 40 and 60 days of treatment in parallel to the observed changes in serum LH, demonstrating that one mechanism by which chronic GnRH analog treatment leads to pituitary desensitization is down-regulation of pituitary GnRH receptors. Testosterone administration alone decreased pituitary GnRH receptor binding. Combined GnRH analog and testosterone administration prevented the increase in pituitary GnRH receptors observed with analog administration alone. These studies demonstrate that changes in pituitary GnRH receptor binding correlate with changes in serum LH and that the stimulatory effects of analog administration on LH are sensitive to inhibition by small doses of testosterone.     

When the lead extractor becomes the assistant for new lead implant

With the increasing number of cardiovascular implantable electronic device upgrade and vein obstruction caused by previous leads, it is important to have alternative techniques to upgrade the device with the maintenance of functioning leads.We report an 83-year old male with 13-year old one-lead dual-chamber pacemaker, ischemic cardiac disease and pre-dialytic chronic kidney disease submitted to an upgrade to cardiac resynchronization therapy. A sub-occlusion in the transition of left brachiocephalic vein and the superior vena cava was documented. Re-permeabilization was only achieved with a TightRail? rotating dilator sheath over a guidewire with successful left ventricle lead implant.     

Importance of estrus on pregnancy per insemination in suckled Bos indicus cows submitted to estradiol/progesterone-based timed insemination protocols

The objective was to evaluate the effect of estrus occurrence (based on removal of tail-head marks) on ovarian responses and pregnancy per AI (P/AI; 30 d after AI) in suckled Bos indicus beef cows submitted to timed AI (TAI) protocols. Cows received an intravaginal device containing 1.0 g progesterone, and 2.0 mg estradiol benzoate im; 8 d later, the intravaginal device was removed, and they were given PGF_2α (0.25 mg of cloprostenol sodium) and 300 IU of eCG, with TAI 48 to 52 h later. In Experiment 1, cows were assigned to receive one of three treatments: 1 mg of estradiol cypionate (ECP) im at progesterone (P4) device removal (N = 178); 10 μg of GnRH im at TAI (N = 190); or both treatments (N = 172). In cows given estradiol (ECP or ECP + GnRH), more displayed estrus (P = 0.002) and became pregnant (P < 0.0001) compared with those receiving only GnRH. In Experiment 2, the effect of the occurrence of estrus on ovarian responses was evaluated in cows (N = 53) synchronized using ECP at device removal. Cows that displayed estrus had a greater diameter of the largest follicle (LF) at device removal (P < 0.0001), a greater diameter at TAI (P < 0.0001), a greater ovulation rate (P = 0.02), a larger CL (P = 0.02), and a greater P4 concentration (P < 0.0001) than cows that did not display estrus. In Experiment 3, the effect of GnRH treatment on P/AI at TAI was evaluated in cows that received ECP at device removal, and either displayed, or did not display, estrus (N = 726). There was no estrus by GnRH interaction (P = 0.22); the P/AI was greater (P < 0.0001) in cows that displayed estrus (61.9%) than cows that did not display estrus (41.4%). However, GnRH did not improve (P = 0.81) P/AI (GnRH = 53.7% vs. no GnRH = 52.6%). In conclusion, exogenous estradiol at device removal increased both the proportion of suckled Bos indicus cows that displayed estrus and P/AI. Cows that displayed estrus had better ovarian responses (i.e., larger follicles at TAI, a greater ovulation rate, larger CL, and greater P4 concentrations) following an estradiol/P4-based synchronization protocol. Although occurrence of estrus improved pregnancy outcomes, GnRH at TAI did not improve P/AI in suckled Bos indicus cows treated with ECP, regardless of estrus occurrence.     

Implantable microfluidic device for the formation of three-dimensional vasculature by human endothelial progenitor cells

Vasculogenesis is an important morphogenetic event for vascular tissue engineering and ischemic disease treatment. Stem and progenitor cells can contribute to vasculogenesis via endothelial differentiation and direct participation in blood vessel formation. In this study, we developed an implantable microfluidic device to facilitate formation of three-dimensional (3D) vascular structures by human endothelial progenitor cells (hEPCs). The microfluidic device was made of biodegradable poly(lactic-co-glycolic acid) (PLGA) using a microchannel patterned silicon wafer made by soft lithography. A collagen type I (Col I) hydrogel containing hEPCs filled the microfluidic channels to reconstitute a 3D microenvironment for facilitating vascular structure formation by hEPCs. The device seeded with hEPCs was implanted into the subcutaneous space of athymic mice and retrieved one and four weeks after implantation. Histology and immunohistochemistry revealed that hEPCs formed a 3D capillary network expressing endothelial cell-specific proteins in the channel of the PLGA microfluidic device. This result indicates that a 3D microscale extracellular matrix reconstituted in the microchannel can promote the endothelial differentiation of hEPCs and in turn hEPC-mediated vasculogenesis. The PLGA microfluidic device reported herein may be useful as an implantable tissue-engineering scaffold for vascularized tissue reconstruction and therapeutic angiogenesis.     

A Multimodal,SU-8 - Platinum - Polyimide Microelectrode Array for Chronic In Vivo Neurophysiology

Utilization of polymers as insulator and bulk materials of microelectrode arrays (MEAs) makes the realization of flexible, biocompatible sensors possible, which are suitable for various neurophysiological experiments such as in vivo detection of local field potential changes on the surface of the neocortex or unit activities within the brain tissue. In this paper the microfabrication of a novel, all-flexible, polymer-based MEA is presented. The device consists of a three dimensional sensor configuration with an implantable depth electrode array and brain surface electrodes, allowing the recording of electrocorticographic (ECoG) signals with laminar ones, simultaneously. In vivo recordings were performed in anesthetized rat brain to test the functionality of the device under both acute and chronic conditions. The ECoG electrodes recorded slow-wave thalamocortical oscillations, while the implanted component provided high quality depth recordings. The implants remained viable for detecting action potentials of individual neurons for at least 15 weeks.     

Tissue engineering of a bioartificial kidney

Tissue engineering is a rapidly growing field in biotechnology. The use and packaging of synthetic materials, biologic compounds, and cellular components of specific tissues can be envisioned to replace physiologic function of diseased organs. Long-term ex vivo therapy for kidney failure has been achieved, so that the kidney may be the first solid organ in which tissue engineering concepts can produce an implantable device for long-term in vivo replacement therapy. To replace the kidney's excretory function, an implantable bioartificial kidney requires both a device to replace blood ultrafiltration performed by renal glomeruli and a device to replace transport regulatory function of the renal tubule. The initial concepts for these devices are just beginning to be considered and developed. (c) 1994 John Wiley & Sons, Inc.     

A Fully Implantable Pacemaker for the Mouse: From Battery to Wireless Power

Animal models have become a popular platform for the investigation of the molecular and systemic mechanisms of pathological cardiovascular physiology. Chronic pacing studies with implantable pacemakers in large animals have led to useful models of heart failure and atrial fibrillation. Unfortunately, molecular and genetic studies in these large animal models are often prohibitively expensive or not available. Conversely, the mouse is an excellent species for studying molecular mechanisms of cardiovascular disease through genetic engineering. However, the large size of available pacemakers does not lend itself to chronic pacing in mice. Here, we present the design for a novel, fully implantable wireless-powered pacemaker for mice capable of long-term (>30 days) pacing. This design is compared to a traditional battery-powered pacemaker to demonstrate critical advantages achieved through wireless inductive power transfer and control. Battery-powered and wireless-powered pacemakers were fabricated from standard electronic components in our laboratory. Mice (n = 24) were implanted with endocardial, battery-powered devices (n = 14) and epicardial, wireless-powered devices (n = 10). Wireless-powered devices were associated with reduced implant mortality and more reliable device function compared to battery-powered devices. Eight of 14 (57.1%) mice implanted with battery-powered pacemakers died following device implantation compared to 1 of 10 (10%) mice implanted with wireless-powered pacemakers. Moreover, device function was achieved for 30 days with the wireless-powered device compared to 6 days with the battery-powered device. The wireless-powered pacemaker system presented herein will allow electrophysiology studies in numerous genetically engineered mouse models as well as rapid pacing-induced heart failure and atrial arrhythmia in mice.     

Ovulation, ovarian function, and reproductive performance after treatments with GnRH in postpartum suckled cows

Two experiments were conducted to determine whether treatments with gonadotropin releasing hormone (GnRH) during the early postpartum period in suckled cows would induce ovulation and initiate regular estrous cycles. In Experiment I, 0, 100 or 200mug of GnRH was given to 22 suckled Angus x Holstein cows at three and again at five weeks postpartum. Serum luteinizing hormone (LH) responses did not differ between cows given 100 or 200mug of GnRH. Treatment with GnRH tended to increase the percentage of cows exhibiting estrus by 30 and 60 days postpartum, but reproductive performance during the breeding season did not differ among groups. In Experiment II, 70 suckled Hereford cows were given either no treatment or 200mug of GnRH at 7 weeks postpartum. Cows given GnRH received either no treatment prior to GnRH or were separated from their calves for 24 hr prior to GnRH treatment. Half of the cows that were separated from their calves also received progesterone via a progesterone intravaginal device (PRID) for 12 days prior to calf removal. Treatment with GnRH alone tended to increase the percentage of anestrous cows which ovulated by 8 days after treatment. Calf removal did not increase the ovulatory response to GnRH, but PRID treatment did. More estrous periods were detected in GnRH-treated cows than in control cows during 20 days after GnRH treatment.     

Effect of timing of prostaglandin administration, controlled internal drug release removal and gonadotropin releasing hormone administration on pregnancy rate in fixed-time AI protocols in crossbred Angus cows

Two experiments were conducted to investigate the effects of timing of prostaglandin F2(alpha) (PGF2(alpha)) administration, controlled internal drug release device (CIDR) removal and second gonodotropin releasing hormone (GnRH) administration on the pregnancy outcome in CIDR-based synchronization protocols. In Experiment 1, suckled Angus crossbred beef cows (n = 580) were given 100 microg of GnRH+a CIDR on Day 0. Cows in Group 1 (modified Ovsynch-P) received 25 mg of dinoprost (PGF2(alpha)) and CIDR device removal on Day 8 (AM), 100 microg of GnRH 36 h later on Day 9 (p.m.), and fixed-time AI (FTAI) 16 h later on Day 10 (47.5+/-1.1 h after PGF2(alpha)). Cows in Group 2 (Ovsynch-P) received 25mg of PGF2(alpha) and CIDR device removal on Day 7 (p.m.), 100 microg of GnRH 48 h later on Day 9 and FTAI 16 h later on Day 10 (66.6+/-1.2 h after PGF2(alpha)). Pregnancy rates were 56.5% (170/301) for Group 1 and 55.6% (155/279) for Group 2, respectively (P = 0.47). In Experiment 2, beef cows (n=734) were synchronized with 100 microg of GnRH+CIDR on Day 0, 25 mg of PGF2(alpha) and CIDR device removal on Day 7 and either 100 microg of GnRH 48 h later on Day 9 (Ovsynch-P) and FTAI 16 h later on Day 10 (64.9+/-3.3 h from PGF2(alpha)) or 100 microg of GnRH on Day 10 (CO-Synch-P) at the time of AI (63.2+/-4.2 h from PGF2(alpha)). Pregnancy rates were 48.8% (180/369) for Ovsynch-P and 44.7% (163/365) for CO-synch-P groups, respectively (P = 0.11). In both experiments, there was a locationxtreatment interaction (P<0.05); pregnancy rates between locations were different (P < 0.05) in the Ovsynch-P group. In conclusion, in a CIDR-based Ovsynch synchronization protocol, delaying administration of prostaglandin and CIDR removal by 12 h, or timing of the second GnRH by 16 h, did not affect pregnancy rates to FTAI. Therefore, there may be an opportunity to make changes in synchronization protocols with out adversely affecting FTAI pregnancy rates.     

Method for measuring long-term function of muscle-powered implants via radiotelemetry.

Long-term remote monitoring of muscle-powered implants has been made possible with development of an adjustable workload that can be remotely monitored to assess device function. This technique obviates the need for percutaneous access lines and allows test animals to remain untethered, eliminating deleterious effects caused by infection, sedation, or animal stress. Hardware components include a latex bladder fixed within a hermetically sealed canister, multichannel implantable telemetry unit, and subcutaneous access port (for pressure charge adjustment). To validate this method, in vitro tests were performed by using a third-generation muscle energy converter designed to function as an implantable hydraulic pump. Two channels of telemetered pressure data were collected and used to calculate six indexes of device function. Calculated parameters were then compared with measured values to determine accuracy. Correlation between measured and calculated parameters was high in all instances, with most estimates yielding errors of <3%. These results demonstrate the utility of this approach and support its use as a means to monitor muscle-powered devices during long-term animal trials.     

A CIDR-based timed AI protocol can be effectively used for dairy cows with follicular cysts

The present study evaluated whether a controlled internal drug release (CIDR)-based timed AI (TAI) protocol could be used as an efficient tool for the treatment of ovarian follicular cysts in lactating dairy cows. In the first experiment, lactating dairy cows diagnosed with follicular cysts were randomly assigned to two treatments: (1) a single injection of GnRH at diagnosis (Day 0) and AI at estrus (AIE) within 21 days (GnRH group, n=70), or (2) insertion of a CIDR device containing progesterone and an injection of GnRH on Day 0, PGF(2alpha) injection at the time of CIDR removal on Day 7, GnRH injection on Day 9, and TAI 16h after the GnRH injection (CIDR-based TAI group, n=65). Conception rate after the CIDR-based TAI protocol (52.3%) was greater (P<0.05) than that after AIE following a single GnRH injection (26.9%). In the second experiment, lactating dairy cows diagnosed with follicular cysts (Cyst group, n=16) and cows having normal estrous cycles (CYC group, n=15) received the same treatment: a CIDR device containing progesterone and an injection of GnRH on Day 0, PGF(2alpha) injection at the time of CIDR removal on Day 7, and GnRH injection on Day 9. The proportion of cows with follicular wave emergence and the interval from treatment to follicular wave emergence did not differ (P>0.05) between groups. The mean diameters of dominant follicles on Days 4 and 7 as well as preovulatory follicles on Day 9, and the synchrony of ovulation following the second injection of GnRH did not differ (P>0.05) between groups. These data suggest that the CIDR-based TAI protocol results in an acceptable conception rate in dairy cows with follicular cysts.     

Review of techniques for diagnosis of catheter-related Candida biofilm infections

Recent evidence shows that many hospital-acquired infections, including most device-associated infections, involve the persistence of sessile organisms in the form of biofilms that are attached to a device surface and encased in an extracellular matrix. The cells in this environment exhibit an altered phenotype with respect to antimicrobial resistance and thus are extraordinarily difficult to eradicate without device removal. Although a number of implantable and topical devices are at risk for Candida biofilm formation, this review focuses on the diagnosis of the most common of these infections, biofilm growth on the surface of central venous catheters and urinary catheters.     

High capacity implantable data recorders: system design and experience in canines and Denning black bears

BACKGROUND: Implantable medical devices have increasingly large capacities for storing patient data as a diagnostic aid and to allow patient monitoring. Although these devices can store a significant amount of data, an increased ability for data storage was required for chronic monitoring in recent physiological studies. METHOD OF APPROACH: Novel high capacity implantable data recorders were designed for use in advanced physiological studies of canines and free-ranging black bears. These hermitically sealed titanium encased recorders were chronically implanted and programmed to record intrabody broadband electrical activity to monitor electrocardiograms and electromyograms, and single-axis acceleration to document relative activities. RESULTS: Changes in cardiac T-wave morphology were characterized in the canines over a 6 month period, providing new physiological data for the design of algorithms and filtering schemes that could be employed to avoid inappropriate implantable defibrillator shocks. Unique characteristics of bear hibernation physiology were successfully identified in the black bears, including: heart rate, respiratory rate, gross body movement, and shiver An unanticipated high rejection rate of these devices occurred in the bears, with five of six being externalized during the overwintering period, including two devices implanted in the peritoneal cavity. CONCLUSIONS: High capacity implantable data recorders were designed and utilized for the collection of long-term physiological data in both laboratory and extreme field environments. The devices described were programmable to accommodate the diverse research protocols. Additionally, we have described substantial differences in the response of two species to a common device. Variations in the foreign body response of different mammals must be identified and taken into consideration when choosing tissue-contacting materials in the application of biomedical technology to physiologic research.     

一种新型植入式装置无线数据传输方法

提出了一种植入式装置无线数据传输方法,以射频电磁波作为信息传输媒介,实现体内植入式装置与体外程控仪的双向通信。文中提出的＂b it-by-b it＂遥测方式可以显著降低起搏器等植入式装置的功耗,延长其使用寿命。     

Pituitary gonadotropin-releasing hormone receptor content in rats with polycystic ovaries

A single injection of estradiol valerate (EV) induces, after a lag period of 4-6 wk, a chronic anovulatory polycystic ovarian (PCO) condition in adult rats. This condition is associated with a selective compromise of luteinizing hormone (LH) release and/or synthesis reflected in low basal serum LH concentrations, decreased pituitary content of LH, and decreased gonadotropin-releasing hormone (GnRH)-stimulated LH secretion. The present study was undertaken to determine to what extent the aberrant LH release in rats with PCO could be related to alterations in pituitary content of GnRH receptors. Pituitary GnRH-receptor content was assessed by the evaluation of saturation binding of a GnRH analog, [125I]-D-Ala6-des-Gly10-GnRH, to pituitary membrane preparations. The receptor content of pituitaries from rats with PCO was compared to that obtained from intact animals at estrus and diestrus. Receptor levels in ovariectomized normal rats and rats with PCO were also assessed. The pituitary GnRH receptor content in PCO rats was similar to that observed in normal controls at estrus and was significantly lower than that for rats at diestrus. Although a twofold increase in pituitary GnRH receptor content was observed at 28 days following the castration of control rats, GnRH receptor content in the pituitaries of PCO rats, at 28 days following ovariectomy, remained unchanged. Although, castration-induced elevations in mean serum LH and follicle-stimulating hormone (FSH) concentrations were observed in both the PCO and control animals, the rise in both gonadotropins was significantly attenuated in the PCO-castrates when compared to the ovariectomized controls. Since GnRH is a major factor in the regulation of pituitary GnRH receptor content, these findings suggest that hypothalamic GnRH release is impaired in rats with PCO and that this impairment is independent of any influences from the polycystic ovaries.     

Time-related neuroendocrine manifestations of puberty: a combined clinical and experimental approach extracted from the 4th Belgian Endocrine Society lecture

The neuroendocrine manifestations of puberty converge on changes in GnRH secretion. Their appraisal through the assay of GnRH-like material in 24-hour urine extracts shows an increased excretion of this material in the late prepubertal period. The most striking pubertal changes in GnRH secretion occur on a circadian and ultradian basis. In man, they can be evaluated only indirectly. The circadian variations in LH and FSH secretion characteristic of puberty may be observed in timed fractions of 24-hour urine with some delay when compared to the variations of plasma levels. Studies on the frequency of pulsatile LH secretion and during chronic intermittent administration of GnRH support the existence of an increased frequency of GnRH secretory episodes at puberty. LH response to synthetic GnRH is directly related to the frequency of stimulation by endogenous GnRH pulses and provides a very useful index of neuroendocrine maturation in patients with delayed or precocious puberty. A direct evaluation of pulsatile GnRH secretion is possible using the rat hypothalamus in vitro. In these experimental conditions, the frequency of pulsatile GnRH release increases during very early stages of sexual maturation in the male rat. GnRH itself and beta-endorphin are inhibitory regulators of GnRH secretion in vitro and may participate in the mechanisms restraining the pulse-generating machinery in the hypothalamus before puberty.     

Programmed electrical stimulation and Amiodarone therapy for the control of persistent junctional tachycardia

The use of radiofrequency as a means of synchronization and stimulation does not necessitate an external lead, and thus has allowed the construction of an implantable device for long-term treatment of reentry tachycardias. The device is used along with Amiodarone therapy and can be triggered by the patient himself.     

A Canadian hospital's experience with the automatic implantable cardioverter/defibrillator.

The automatic implantable cardioverter/defibrillator is a device that can be implanted in patients for treatment of recurrent ventricular tachycardia and ventricular fibrillation. It was recently approved for clinical use in Canada. The authors describe their experience with 12 patients (mean age 51.3 years) who underwent implantation of a defibrillator. All 12 patients had a history of documented ventricular fibrillation, which was idiopathic in 3 and due to ischemic heart disease in 9. Electrophysiologic testing revealed inducible ventricular tachycardia or ventricular fibrillation in 8 of the 10 patients tested. An important criterion for selection for implantation was failure of pharmacologic therapy to suppress ventricular arrhythmias induced during electrophysiologic testing. Of the 12 patients, 1 died within 24 hours after implantation. During a mean follow-up period of 15.5 months there were no further deaths. All the surviving patients expressed satisfaction with the device; five of the seven under the age of 60 years have returned to work, and one has returned to school. This initial favourable experience with the automatic implantable cardioverter/defibrillator suggests that future increases in the availability of the device and improvements in its function will lead to much more widespread use, as the population of patients at risk of sudden cardiac death is large.