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Resumen Introduccion Los programas de recuperacion precoz (rapid recovery, RP) en artroplastia total de rodilla pueden mejorar la funcionalidad a la vez que se reducen los costes. El objetivo del estudio es comparar los resultados de un programa de rehabilitacion precoz con nuestro protocolo habitual. Pacientes y metodos Se realizo un ensayo clinico aleatorizado (NCT03823573) en pacientes operados de artroplastia total de rodilla. El grupo intervencion (RP) recibio infiltracion periarticular con levobupivacaina e inicio deambulacion supervisada a las 4-6 h tras la intervencion. El grupo control (C) empleo drenaje y recibio un bloqueo femoral e inicio la deambulacion al retirar el drenaje. Los pacientes completaron un cuestionario Oxford Knee Score preoperatorio y a los 6 meses. La incidencia de trombosis venosa profunda asintomatica se analizo mediante ecodoppler. El seguimiento minimo fue de 6 meses. Resultados Fueron incluidos 175 pacientes (92 pacientes en el grupo C y 83 en el RP). No hubo diferencias en sexo, edad, tipo de protesis, descenso de hemoglobina, necesidad de transfusiones, balance articular activo al alta (C: 82,6°; RP: 85°) ni al finalizar el seguimiento (C: 105,1°; RP: 106,6°), mejoria del cuestionario (C: 17,5 puntos; RP: 19,3 puntos), satisfaccion del paciente o reatenciones hospitalarias (C: 7,6%; RP: 10,8%). Se observo significacion en el tiempo de isquemia (C: 81,29 min; RP: 85,35 min; p = 0,03), necesidad de rescate con opioides (C: 19,7%; RP: 38,6%; p = 0,007), estancia media (C: 3,84 dias; RP: 2,54 dias; p Conclusion El protocolo RP puede reducir la estancia hospitalaria sin aumentar las complicaciones ni las reatenciones.

Resumen Introduccion La practica en modelos experimentales es una opcion valida que mejora los resultados y acorta las curvas de aprendizaje de las tecnicas quirurgicas. Nuestro objetivo fue desarrollar un modelo en plastico, impreso en 3D para la docencia, el entrenamiento y la formacion en ureteroscopia flexible, analizando costes e idoneidad para la practica de esta tecnica quirurgica. Metodos Se elaboro un modelo impreso en 3D a partir de una tomografia axial computarizada de una via urinaria superior de un paciente real. La segmentacion se llevo a cabo mediante el software HorosTM y la impresion mediante una impresora FDM-Ultimaker. Se numero los calices renales para ser identificados, como en el plan de formacion de tratamiento endoscopico de litiasis, ejercicio 4, de la Asociacion Europea de Urologia. Se utilizaron: un ureteroscopio flexible desechable Innovex (Palex) y cestillas de nitinol (Coloplast). Resultados El tiempo de impresion fue de 19 h, con un coste total de 8,77 €. El modelo tridimensional permitio la introduccion del ureteroscopio flexible y la exploracion de los calices renales por parte de urologos tanto en formacion como en ejercicio actual de la especialidad sin dificultad. El modelo tambien permitio la utilizacion de cestillas y la movilizacion y extraccion de litiasis previamente colocadas. Conclusion Damos a conocer un modelo tridimensional valido para ejercicios de formacion en ureteroscopia flexible con unos costes razonables, que permitira adquirir la destreza y la confianza necesaria para iniciar el procedimiento en un escenario real.

Introduction In-hospital 3D printing is being implemented in orthopaedic departments worldwide, being used for additive manufacturing of fracture models (or even surgical guides) which are sterilized and used in the operating room. However, to save time and material, prints are nearly hollow, while 3D printers are placed in non-sterile rooms. The aim of our study is to evaluate whether common sterilization methods can sterilize the inside of the pieces, which would be of utmost importance in case a model breaks during a surgical intervention. Material and method A total of 24 cylinders were designed and printed with a 3D printer in Polylactic Acid (PLA) with an infill density of 12%. Manufacturing was paused when 60% of the print was reached and 20 of the cylinders were inoculated with 0.4 mL of a suspension of S epidermidis ATTCC 1228 in saline solution at turbidity 1 McFarland. Printing was resumed, being all the pieces completely sealed with the inoculum inside. Posteriorly, 4 groups were made according to the chosen sterilization method: Ethylene Oxide (EtO), Gas Plasma, Steam Heat or non-sterilized (positive control). Each group included 5 contaminated cylinders and 1 non-contaminated cylinder as a negative control. After sterilization, the inside of the cylinders was cultured during 7 days. Results We observed bacterial growth of just a few Forming Colony Units (FCU) in 4 out of 5 positive controls and in 2 out of 5 contaminated cylinders sterilized with Gas Plasma. We could not assess any bacterial growth in any of the EtO or Steam Heat samples or in any of the negative controls. Pieces sterilized under Steam Heat resulted completely deformed. Conclusions High temperatures reached during the procedure of additive manufacturing can decrease the bacterial load of the biomodels. However, there is a potential risk of contamination during the procedure. We recommend sterilization with EtO for in-hospital 3D-printed PLA hollow biomodels or guides. Otherwise, in case of using Gas Plasma, an infill of 100% should be applied.

Objectives We present the results of the prospective follow up of a sample of large head metal-metal total hip arthroplasty obtained after the safety alert regarding a higher incidence of revision of these implants. Material and methods All patients implanted with the Recap-M2a-Magnum cup between 2008 and 2011 were included. They were prospectively reviewed recording Harris Hip Score, clinical symptoms of chromium or cobalt intoxication. Serum levels of these ions were requested as well as X-rays and ultrasonography. An MRI was performed in the cases of positive ultrasonography. Results Twenty-six males with a mean age of 48.54 years [32–62, SD: 7.18] were included. An anterolateral approach and Bi-Metric (7) and F-40 (19) stems were used. Cephalic diameters ranged 42–52 (mode: 46) and the mean cup inclination was 39.35° [21–59°, SD: 9.78]. During follow-up (7.3 years [5.9–9.4; SD: .78]), 3 patients (11.5%) underwent revision (2 cases aseptic loosening, 1 pseudotumour). Mean time until revision was 5.4 years [3.1–8.0; SD: 2.48]. The accumulated survival probability was 88.5% (95% CI 76.3–100%). Harris Hip Score was 94.47 [66.5–100; SD: 8.94] and the patients showed no metallic intoxication symptoms. The levels of chromium were 1.88 μg/dl [0.6–3.9] and cobalt 1,74 μg/dl [0.5–5,6]. One pseudotumour was found in an asymptomatic patient, and small amounts of periarticular liquid were found in 5 patients (19.2%) Discussion and conclusions High revision rates are still found when follow up is extended due to aseptic loosening and pseudotumour formation. MRI might not be the most adequate test to study the complications of these prostheses.

Objectives Our aim was to investigate whether patients presenting with fragility fractures of the proximal femur are receiving osteoporosis treatment and to assess the number of other fragility fractures they have sustained prior to admission. Methods All patients presenting to our institution with fragility fractures of the proximal femur within an 18-month period (January 2012–August 2013) were included. Patient demographics; fracture classification (AO/OTA); American Society of Anesthesiologists (ASA) grade; Abbreviated Mental Test Score (AMTS) on admission; type of operation; time to operation; peri -operative complications; length of hospital stay (LOS); walking status; osteoporotic medication; Dual-energy X-ray absorptiometry (DEXA) results; additional fragility fractures; and mortality were collected and analysed. Results A total of 1004 patients (278 male) met the inclusion criteria and were included into the study. The mean age was 82.01 years and mean LOS was 19.54 days. Fifty-four per cent of the patients were admitted from their own homes whereas 43% were capable to walk indoors without any aids before their injury. Mean time to surgery was 2.06 days (Median: 1.31, range: 0–26 days). Three hundred and six patients (30.5%) had at least another fragility fracture before the index episode (mean 1.40 fractures; SD: 0.71 fractures; range: 1–6 fractures). Only 16.4% were under complete osteoporosis treatment on admission, defined as receiving calcium with vitamin D and a bisphosphonate or an alternative agent. When we compared patients without a history of a previous fragility fracture (Group A) and patients with at least another previous fragility fracture (Group B), we found that patients in Group B had a significantly lower AMTS score, lower bone mineral density (BMD) as evident on the DEXA scan, an inferior mobility before admission and a higher incidence of extracapsular fractures (p On discharge, patients in Group B had a higher chance of receiving complete bone protection compared to group A (27.9% versus 41.7%; p Following discharge, 11.2% of the patients sustained an additional fragility fracture. The mean time from the index episode to the additional fracture was 0.65 years, whilst these injuries were more frequent in Group B (RR = 1.638; p Conclusion Patients presenting with a hip fracture are generally under-treated for osteoporosis. Post-operative assessment by a designated geriatrician and use of a standardised protocol is of paramount importance for reducing the risk of additional fragility fractures. Additionally, screening of the elderly population for identifying the patients who suffer from osteoporosis can potentially reduce the risk of sustaining a further fragility fracture.

Distal radius fracture (DRF) is the most common upper extremity fracture. The incidence of complications after surgical treatment still remains high. The objective of our study was to assess functional and radiological results of DRF treated with volar locking plate.We conducted a retrospective study including DRF treated by open reduction and internal fixation with a volar locking plate during a period of 8 years (2010-2018). Data were collected from clinical records and included patient demographics, fracture characteristics, radiological parameters (radial inclination, palmar tilt, ulnar variance, articular step-off), range of motion (ROM), complications and reinterventions.A total of 170 patients (63.5% female) met the inclusion criteria with a mean age of 55.9 years [24.1-83.7; 13.76]. High energy injuries were more frequent in younger patients (25.56% vs 3.95%; p 0.0001). AO classification was: 2R3A: 23.5%, 2R3B: 24.1% and 2R3C: 52.4%. Time until surgery was 8.5 days, longer in extraarticular fractures (AO-2R3A: 12.22 days, AO-2R3B: 7.97 days, AO-2R3C: 7.04 days, p = 0.018). Younger patients had better radial inclination (excellent in 85.1% vs 59.21%, p = 0.001) and radial shortening (excellent in 100% vs 88.15%, p = 0.001). ROM was: flexion 63° [0-90°; 19.73], extension 57.67° [5-90°; 21.61], pronation 78.94° [40-90°] and supination 81.76° [38-90°; 8.87]. We found a weak correlation between ROM in flexion-extension and radial shortening (r = 0.218; p = 0.001) and articular step-off (r = -0.269; p = 0.002). We had 39 complications (21.18% of patients). 10.6% of patients needed a re-operation, being the most frequent hardware removal (6.5%) and tendinous disruptions repairs (2.4%). Reintervention rate was higher in younger patients (14.9% vs 5.3%; p = 0.042), complete articular fractures (AO-2R3A: 2.5%; AO-2R3B: 7.3%; AO-2R3C: 15.7%; p = 0.037) and high energy injuries (8.4% vs 22.2%; p = 0.044). Patients undergoing reintervention had a decreased flexion-extension (94.44° vs 123.83°, p = 0.007).Radial shortening and articular step-off seem the most important predictors for postoperative range of motion. Worse radiological outcomes are observed in complete articular fractures and those affecting elder population. Although radiological and functional results are good or excellent in most of cases, the incidence of complications and need for reintervention still remains noticeable. Re-operations were more frequent in younger patients, complete articular fractures and high energy injuries, and it worsened functional outcomes.

IntroductionPatient specific instrumentation is a novel aid the industry has been implementing in our hospitals during the last years. However, the cost of the process and the impossibility to discuss face to face with the manufacturer the needs of the particular cases may favor the in-hospital production of these devices.We present the design and manufacturing process of three patient-specific devices to treat complex common situations in orthopaedic surgery, such as intramedular tumor resections, long bone pseudarthrosis and malunions of articular fractures.Material and MethodsCT scans of the region of interest were performed to all the patients. Horos® software was used to isolate the affected bone region, whereas Meshmixer® software was used to create the patient specific guides. An Ultimaker 2+® 3D printer was used to print the guide, in a biocompatible material (SmartFil® Medical). The printing parameters included a nozzle of 0.6 mm and a layer height of 0.1 mm.There was one case of a low-grade chondrosarcoma in which we created a guide to resect enough cortical bone to make an extended curettage of the lesion. Phenol and PMMA were used as adjuvants after the curettage. We used the same guide to obtain an exact replica of allograft which was later placed in position.We had a femur diaphyseal atrophic pseudarthrosis in a patient with achondroplasia, in which the guide permitted the resection of the affected bone with a saw and posterior osteosynthesis with an anterior LCP plate.Finally, in an intraarticular malunion of a distal radius Die-Punch fracture we used the guide to make an osteotomy of the affected articular portion and correct the articular surface. Bone autograft was added prior to the osteosynthesis with a plate.ResultsMean design time was 6.3 hours. Mean printing time was 5.2 hours. The price of the filament used for each guide was under 10 USD. The mean time from the CT-Scan and the surgery being performed was 2.7 months. In all cases the patient specific guides fitted in the bone and permitted the planned resection/osteotomy.All the surgeries fulfilled their purpose.ConclusionsPatient specific guides are easy to design in a local setting with the aid of free software. Design time still needs dedication although it permits the manufacturing of the guides following surgeon needs. The use of self-designed and printed guides is safe and accurate, with a low cost for the institutions.

PurposeThe medical morbidity and mortality associated with neck of femur fractures is well-documented, whereas there is limited data for patient-reported outcomes. The aim of this study was to characterize the impact of neck of femur fractures on activities of daily living and patient-reported health-related quality of life.MethodsDesign and participants: Multicentric prospective cohort study. Consecutive sample patients with fragility hip fracture over 50years old admitted in 48 hospitals in Spain.Outcomes: daily living activity function (Barthel Index) and health-related quality of life (EQ-5D) pre-fracture, admission to hospital and at 1- and 4-month follow-up post-fracture.Statistics: Barthel and EQ-5D over time are described as mean (SD) and median (interquartile range).ResultsA total of 997 patients were recruited at baseline with 4-month outcomes available for, and 856 patients (89.5%). Barthel Index fell from 78.77 (23.75) at baseline to 43.62 (19.86) on admission to hospital with the fracture. Scores partially recovered to 54.89 (25.40) and 64.09 (21.35) at 1- and 4-month post-fracture, respectively. EQ-5D fell from a median of 0.75 (0.47-0.91) to -0.01 (-0.03 to 0.51) on admission. Partial recovery was observed again to (0.51 (-0.06 to 0.67)) and (0.60 (0.10 to 0.80)) at 1- and 4-month post-fracture, respectively.ConclusionsHip fracture results in a large decline in the ability to perform activities of daily living and patient-reported health-related quality of life with only partial recovery amongst survivors 4-month post-fracture., This work has been sponsored by an unrestricted grant of AMGEN to the Hospital del Mar Institute of Medical Investigation (IMIM).

Introduction and objective Arthrodiastasis consists of the mobilisation of the hip while distracted by a hinged distractor. The aim of our study is to show the results of the treatment of late-onset Perthes disease with this technique in our institution. Material and methods Ambispective review of all the cases that underwent this surgery in our Department of Paediatric Orthopaedics between 2010 and 2014 with the following inclusion criteria: patients over 8 years old, with minimal deformity, and poor prognosis. Mobility, pain, Trendelenburg sign, radiographic sphericity, and complications were assessed. Results A total of 13 patients (9 male, 4 female) were included. The mean age was 11.93 years. Radiographic risk signs were present in 7 cases. The mean decrease in pain measured with a visual analogue scale (VAS) after the surgery was 3.83 points (P Conclusions Arthrodiastasis is a minimally invasive technique, useful in the treatment of late onset Perthes disease. It improves clinical results and maintains cephalic shape when applied in early stages. However, it is not free of complications, which can involve as many as 8 out of 10 patients treated with the technique.

We have characterised 997 hip fracture patients from a representative 45 Spanish hospitals, and followed them up prospectively for up to 4 months. Despite suboptimal surgical delays (average 59.1 hours), in-hospital mortality was lower than in Northern European cohorts. The secondary fracture prevention gap is unacceptably high at 85%. Purpose To characterise inpatient care, complications, and 4-month mortality following a hip or proximal femur fracture in Spain. Methods Design: prospective cohort study. Consecutive sample of patients >= 50 years old admitted in a representative 45 hospitals for a hip or proximal femur fragility fracture, from June 2014 to June 2016 and followed up for 4 months post-fracture. Patient characteristics, site of fracture, in-patient care (including secondary fracture prevention) and complications, and 4-month mortality are described. Results A total of 997 subjects (765 women) of mean (standard deviation) age 83.6 (8.4) years were included. Previous history of fracture/s (36.9%) and falls (43%) were common, and 10-year FRAX-estimated major and hip fracture risks were 15.2% (9.0%) and 8.5% (7.6%) respectively. Inter-trochanteric (44.6%) and displaced intra-capsular (28.0%) were the most common fracture sites, and fixation with short intramedullary nail (38.6%) with spinal anaesthesia (75.5%) the most common procedures. Surgery and rehabilitation were initiated within a mean 59.1 (56.7) and 61.9 (55.1) hours respectively, and average length of stay was 11.5 (9.3) days. Antithrombotic and antibiotic prophylaxis were given to 99.8% and 98.2% respectively, whilst only 12.4% received secondary fracture prevention at discharge. Common complications included delirium (36.1 %) and kidney failure (14.1%), with in-hospital and 4-month mortality of 2.1% and 11% respectively. Conclusions Despite suboptimal surgical delay, post-hip fracture mortality is low in Spanish hospitals. The secondary fracture prevention gap is unacceptably high at > 85%, in spite of virtually universal anti-thrombotic and antibiotic prophylaxis., Funded in part by CIBER on Frailty and Healthy Aging (CIBERFES), Instituto Carlos III, Spanish Ministry of Economy and Competitiveness, FEDER funds. Unrestricted research grant from Amgen S.A. Amgen did not have any role in study design, collection, analysis and interpretation of data, writing the report, and the decision to submit the report for publication.

Introduction Several authors have suggested a correlation between the fracture patterns of proximal femur fractures and the degree of hip osteoarthritis (HOA), but the current evidence to support this are insufficient. The aim of our study was to demonstrate whether there is an association between the grade of HOA and fracture pattern observed, in patients presenting with a fragility fracture of the proximal femur. Materials and methods We contacted a retrospective review of all patients presenting to our institution with fragility fractures involving the proximal femur, between March 2012 and October 2013. Pathological fractures, high-energy injuries and patients with less than one year of follow-up were excluded from further analysis. Admission radiographs and severity of HOA were assessed according to Kellgren and Lawrence scale (minimal: Grades 1–2; severe: Grades 3–4). Fractures were classified according to AO/OTA classification. Results A total of 1003 patients (725 females; 1003 fractures) met the inclusion criteria, having a mean age of 81.5 (46–106 years). With regards to fracture classification, 417 (41.6%) fractures were classified as extracapsular and 586 (58.4%) as intracapsular. A total of 939 (93.9%) patients presented with minimal HOA, whilst 61 (6.1%) of the patients presented with severe HOA. Of the 61 patients presenting with severe HOA, 42 patients (68.9%) sustained a 31A-interthrocanteric fracture and 19 patients (31.1%) sustained a 31B-intracapsular fracture. Regarding the patients presenting with minimal HOA (832 patients in total), 323 patients (38.8%) sustained 31A-intertrochanteric fracture and 509 patients (61.2%) sustained a 31B-intracapsular fracture. Patients presenting with severe HOA were found to have a statistically significant chance to present with an extracapsular fracture (p Conclusions The degree of HOA is related to the fracture pattern in patients presenting following simple mechanical falls. More specifically, higher grades of HOA are associated with extracapsular fracture patterns, whereas lower grades of HOA are associated with intracapsular fracture patterns.

Objective Evaluation of the surgical management, outcome and complications in patients with pertrochanteric fractures treated with PFNA nail. Materials and methods A retrospective study was conducted on 200 patients treated consecutively between April 2010 and February 2012. Radiological assessments were performed before and after the surgery, and during the follow-up (fracture reduction, blade position, consolidation or collapse signs). A clinical evaluation was performed as regards walking capabilities. The results were compared with those of a previous study on 700 patients treated with gamma 3 and TFN nails. Results The blade position was center–center in 64% of patients, and decreased to 53% in the mechanical complications group. Tip-apex distance was less than 25 mm in 91.5%. The average hospital stay was 9.17 days, with a mean post-surgery stay of 5.95 days. Complications (7.5%): 2 cut out (1%), one cut through (0.5%), 4 cases of helical blade sliding (2%), one failure in distal locking procedure (0.5%), 2 cases with painful fasciae latae (1%), one union delay (0.5%), 2 cases of non-union with hardware failure (1%), one case of intense bleeding related to distal locking of the nail (0.5%), and one case of avascular necrosis (0.5%). Conclusions The PFNA helical blade system seems to reduce the incidence of cut out and cut through in osteoporotic bone. Blade position was one of the main parameters associated with mechanical complications.

The coeliac ganglion and the ovary are related by the superior ovarian nerve, which penetrates into the ovary by the hilium and innervates mainly the ovarian stroma. On the other hand, it is known that the gaseous neurotransmitter nitric oxide (NO) and the two isoforms of its synthesis enzyme, the nitric oxide synthetase (NOS), are present in the ovary. Both innervation and NO participate in ovarian steroidogenesis. Therefore, the purposes of this work were (a) to standardize an in vitro coeliac ganglion-superior ovarian nerve-ovary integrated system in prepubertal rats; (b) to determine the presence of NO in the ovary and analyze the ganglionic cholinergic effect on the ovarian release of androstenedione, progesterone and NO; and (c) to assess the steroids/NO relationship. The system was incubated in buffer solution for 120 min, with the ganglion and ovary located in different compartments and linked by the superior ovarian nerve. From the results obtained, it is concluded that the system is viable and functional. The presence of basal NO is stimulated by the cholinergic action, while the release of the steroids is inhibited, which might indicate that the ganglionic cholinergic effect is probably mediated by NO. To our knowledge, this work constitutes the first study of the relationship between the neural cholinergic action and NO on the ovarian steroidogenesis of prepubertal rats.

The coeliac ganglion neurons, whose axons constitute the superior ovarian nerve (SON), contain cholinergic receptors. The aim of this work was to study the effect of cholinergic agents added to the coeliac ganglion on the release of ovarian progesterone in the coeliac ganglion–SON–ovary in vitro system. We also analyzed the release of norepinephrine in the ovarian compartment and its possible relationship with the release of progesterone. After the addition of cholinergic agents in the ganglion compartment, progesterone release was determined by radioimmuneassay (RIA) and norepinephrine by catecholamine assay (HPLC). The release of progesterone and norepinephrine in the ovary compartment was studied during period of 180 min in pre-oestrus (PE), oestrus (E), dioestrus day 1 (D1) and dioestrus day 2 (D2) rats. The most relevant results concerning the action of acetylcholine were found on PE and dioestrus. On PE, the pre-ovulatory peak of progesterone, which is known to respond to the endocrine action, was not modified by neural effect of acetylcholine in our scheme. On the other hand, the progesterone peak occurs in the afternoon of D1, which has been described as independent of the gonadotrophic action but was inhibited by neural effect of acetylcholine in our experimental scheme. This action on D1 was accompanied by a decrease of norepinephrine release in the ovary compartment. We conclude that the action of cholinergic agents varies according to the oestrous cycle stage and constitutes one of the factors governing the secretory activity of the ovarian steroids, in this case, progesterone.

The neural pathway most related with ovarian steroidogenesis has been identified as the superior ovarian nerve (SON). This work constitutes the first study of the effects of early ovarian SON transection, which was performed in rats of 4 days of age (SON-t rats) to magnify the effects of the denervation. The rats were studied at the prepubertal (30 days), peripubertal (41 days) and adult cyclic in dioestrus (60 days) reproductive stages. The SON-t rats showed a delay of vaginal opening, a notable disruption of oestrous cyclicity, and a large number of corpora lutea. In all the stages, the circulating levels of FSH, prolactin and growth hormone were lower in SON-t rats than in controls, whereas LH did not vary. Serum androstenedione levels were higher in SON-t rats at 30 days and lower at 41 days, compared with control animals while no difference was observed at 60 days. Serum progesterone levels did not differ between control and SON-t, but serum oestradiol concentrations were higher in SON-t rats in all of the stages. At the peripubertal stage, there were fewer ovarian beta-adrenergic receptors in SON-t ovaries, associated with a rise in the ovarian content of norepinephrine, but no changes were observed in SON-t rats at 30 and 60 days with respect to the controls. The release of progesterone in vitro from luteal cell in SON-t rats at 60 days was reduced in basal condition and under ovine LH or FSH stimulation, when compared with control animals; while no difference was observed in presence of isoproterenol or androstenedione in the culture medium. In corpora lutea of SON-t rats at 60 days, no change was observed in the activity of 3beta-hydroxysteroid dehydrogenase (3beta-HSD), but the activity of 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) was reduced, suggesting abnormal luteolysis in spite of the large number of corpora lutea. The interruption of innervation at an early age by SON transection is very important in the regulation of ovarian development in prepubertal and cyclic rats. The functional changes observed in the ovary suggest a possible alteration in the hypothalamic-hypophyseal axis.

Strong evidence indicated that spermatic nerves are involved in the regulation of testosterone secretion. Our previous work showed that the inferior spermatic nerves play a more significant role than the superior ones in the regulation of testosterone secretion. However, it is unknown whether traditional neurotransmitters are involved in this regulation. In order to evaluate this point, the present experiments were carried out in an in vitro system where an isolated testis-spermatic nerve plexus preparation was incubated in two separate containers, one for the testis and the other for the nerve plexus and both interconnected by the inferior spermatic nerves. Both tissues were maintained in the same environmental conditions except for the neurotransmitter treatment, applied only to the nerve plexus. Acetylcholine can significantly inhibit the secretion of testosterone until the end of incubation. The present experiments suggest that the secretion of testosterone could be regulated, at least in part, by acetylcholine through the inferior spermatic nerves.

This study investigates the interaction between the effect of epinephrine intracerebroventricular (icv) injection and LH on the progesterone concentration in ovarian vein blood (Po) in vivo, and also, on the release of ovarian progesterone and androstenedione in vitro, in rats on dioestrus day 2. When 2 mg ovine LH were injected in vein (i.v.), Po increased reaching 120+/-12.2 and 151+/-17.5 ng ml(-1) at 22 and 25 min, respectively. Another group of rats was injected intracerebroventricular with 5 microgram epinephrine at time zero, and with 2 mg ovine LH i.v. 3 min later. This time Po decreased during the first 3 min, then increased, reaching 64+/-7.1 ng ml(-1) at 25 min, lower than the Po obtained 22 min after LH i.v. injection only (P

The effect of androstenedione on luteal progesterone production was studied during luteolysis preceding parturition as well as that induced by the antiprogestin RU486 in late pregnant rats. Luteal cells from animals on days 19, 20 or 21 of pregnancy and incubated with 10 microM androstenedione increased progesterone production by 99, 136, and 277%, respectively. The animals receiving androstenedione (10 mg/rat s.c.) on day 19 of pregnancy showed an increase in serum progesterone levels, a decline in luteal 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity and an increase in corpus luteum weight without modifying 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD) activity on day 21 of pregnancy. Androstenedione and testosterone but not dihydrotestosterone were able to prevent the decrease in serum progesterone concentration and corpus luteum weight observed 58 h after treatment with RU486 (2 mg/kg) on day 18 of pregnancy. However, the three androgens studied inhibited the luteal 3 beta-HSD activity but 20 alpha-HSD activity was not affected, when compared with animals receiving RU486 alone. The co-administration of androstenedione with the aromatase inhibitor 4-hydroxyandrostenedione or with the specific antioestrogen ICI 164,384 did not modify the effects induced by androstenedione in RU486-treated rats, indicating that the action of androstenedione on progesterone production and secretion at the time of luteolysis seems to occur through an androgenic mechanism and is not mediated by previous conversion of the androgens to oestrogens. In all experiments the high luteal 20 alpha-HSD activity, that characterizes a luteolytic process, was not modified by androgens. Androstenedione administered to adrenalectomized rats was also able to prevent the decrease in serum progesterone concentration observed in spontaneous or RU486-induced luteolysis. The administration of androstenedione to RU486-treated rats induced a decrease in luteal progesterone content concomitant with an increase in serum progesterone levels. These studies demonstrate that androgens during luteolysis, are able to stimulate luteal progesterone secretion, prevent the loss in corpora lutea weight and enhance the decrease in 3 beta-HSD activity, without affecting the increase in 20 alpha-HSD activity.

To determine if androstenedione, an aromatizable androgen, has a direct effect on luteal progesterone secretion, collagenase-dispersed luteal cells or whole corpora lutea from pregnant rats were incubated in the presence of the androgen. Luteal cells from 15-day pregnant rats responded to androstenedione in a dose-dependent manner, with an increase in progesterone output at doses of 1 and 10 microM, but with no effect at minor doses of the androgen. Luteal cells obtained from animals on day 4, 9, 15 or 19 of pregnancy and incubated with 10 microM of androstenedione, increased progesterone production by 243, 39, 84 and 146%, respectively. Androgens (androstenedione, testosterone or dihydrotestosterone) but no oestrogens (oestradiol or diethylstilboestrol) at a dose of 10 microM, stimulated progesterone production in incubated luteal cells obtained from 15-day pregnant rats. The time-course pattern of androstenedione-induced progesterone production was studied by superfusion experiments using corpora lutea from rats on day 15 of pregnancy. A significant progesterone output was observed when androstenedione, but not oestradiol, was perfused through the luteal tissue. Intrabursal ovarian administration of androstenedione (10 microM) to 19-day pregnant rats induced a significative increase in serum progesterone levels 8 and 24 h after treatment. These in vivo results confirm the stimulatory effect of androstendione on progesterone production obtained in incubated luteal cells from pregnant rats. This study reports a direct luteotrophic effect of androstenedione in rat corpus luteum, not mediated by previous conversion to oestrogens.

Most of the fibres that constitute the superior ovarian nerve (SON) originate at the neuronal bodies of the coeliac ganglion and innervate rat ovarian stroma cells. The purpose of this work was to study the part played by innervation on ovarian release of progesterone on day 15 and at the end of pregnancy in an integrated in vitro system known as the coeliac ganglion-SON-ovary system. We also investigated, in the same system, whether there is some kind of inter-relationship between the effect of adrenergic agents and LH on progesterone release on day 15 of pregnancy. The coeliac ganglion and the ovary were incubated in separate compartments, linked by the SON. The ovary was immersed in 2 ml buffer solution (ovarian compartment) and the coeliac ganglion was immersed in 2 ml of a different buffer solution (ganglion compartment). Under these conditions, the accumulation of progesterone in the ovarian compartment medium was used as an endpoint. Conditions were standardised on day 15 of pregnancy, when the decrease in the release of ovarian progesterone caused by non-specific stimulation on the ganglion with KCl (56 mM) demonstrated the functional integrity of the system. Neural influence was evaluated by the addition of adrenergic agents at a concentration of 10(-6)M to the coeliac ganglion. On day 15 of pregnancy, noradrenaline and propranolol increased progesterone release while phentolamine diminished it. The existence of ganglionic tone was assessed by analysing progesterone basal levels at different stages of pregnancy. The highest secretion of progesterone was found to take place on day 15, diminishing as pregnancy advanced. In addition, adrenergic neural participation was studied during the physiological luteolysis occurring at the end of pregnancy. Major findings were that noradrenaline increased ovarian accumulation of progesterone on day 19 and decreased it on day 20, while propranolol and phentolamine diminished progesterone release on both days. In additional studies, some neuroendocrine aspects were investigated at a peripheral level. The addition of LH only to the ovarian compartment did not affect progesterone secretion. However, when LH in the ovarian compartment was accompanied by noradrenaline, propranolol or phentolamine in the ganglion compartment, the release of progesterone decreased. It can be concluded that modifications of the neural state of the coeliac ganglion affect ovarian progesterone secretion and the physiology of pregnancy via the SON. The results may confirm that the coeliac ganglion-SON-ovary system provides a direct link between the autonomic nervous system and physiological events during pregnancy.

The superior ovarian nerve (SON) arrives at the ovary through the suspensory ligament and innervates mainly the ovarian stroma. Most neurones from which the SON fibres originate are located in the complex coeliac and mesenteric ganglia. Taking into account that other ganglia have been shown to have alpha- and beta-adrenergic receptors, and that the coeliac ganglion receives adrenergic fibres from other sympathetic paravertebral and preaortic ganglia, we utilised adrenergic agonists and antagonists specific to the ganglion, to analyse the role of the alpha and beta receptors in ovarian physiology. To that end, it was necessary to develop and standardise an in vitro coeliac ganglion-SON-ovary (coeliac ganglion-SON-O) experimental system that would enable study of the release of steroids in the ovary in the absence of humoral factors. We investigated the effect of adrenergic agents on the liberation of progesterone in the different stages of the oestrous cycle.To this end we placed the coeliac ganglion and the ovary in different compartments, connected through the SON, to produce a system being studied as a whole. Combined neural and hormonal (luteinising hormone (LH)) effects were also examined. Non-specific stimulation with KCl in the ganglion compartment evoked different responses in terms of release of progesterone, depending on the physiological conditions of the cycle; this demonstrated the sensitivity and viability of the system. During pro-oestrus, stimulation of the ganglion compartment with adrenergic agents such as the agonist noradrenaline or the beta-adrenergic antagonist propranolol, did not modify the release of progesterone. In contrast, the alpha-adrenergic antagonist, phentolamine, induced a strong inhibitory response. During the oestrous stage, noradrenaline was inactive, but phentolamine and propranolol exerted a strong stimulus throughout the experiment. On dioestrus day 1 (D1), both noradrenaline and propranolol increased the release of ovarian progesterone, whereas phentolamine had the opposite effect. Finally, on dioestrus day 2 (D2), what was noteworthy was the pronounced inhibitory effect of noradrenaline, whereas phentolamine was inactive and propranolol showed its greatest stimulatory effect. In order to compare the combined neural and endocrine effects on the ovarian release of progesterone, the experiment was carried out during stages D1 and D2, when the corpora lutea are at their peak of activity. Adrenergic agents were added to the ganglion and LH in a final concentration of 50 ng/ml was added to the ovarian comparment. Different effects were observed indicating a differential response to these agents in stimulated and basal conditions. We conclude that the in vitro coeliac ganglion-SON-ovary system is a functional entity because it possesses its own autonomic tone. This is verified because different basal values of progesterone appear in the different stages of the oestrous cycle. In similar fashion, variations of progesterone induced via the neural pathway were observed under different experimental conditions. In contrast, on D2, noradrenaline added to the ganglion compartment had an inhibitory effect on the liberation of ovarian progesterone.This would indicate that, during this phase, noradrenaline may not be the neurotransmitter released in the ovarian compartment, but that other inhibitory molecules might participate in the observed effects. Finally, during D2, the neural input would condition the ovarian response to LH, facilitating the decrease in progesterone necessary to start a new cycle. The experimental scheme is, in our opinion, a valuable tool for the study of peripheral neural participation in ovarian physiology.

Infection with human retroviruses other than HIV in Spain has only been reported in isolated cases in African immigrants and intravenous drug abusers infected with HTLV-I. The status of the prostitute population is unknown. The sera of 88 prostitutes in Seville were therefore tested for HIV, HTLV-I and HTLV-II, and relevant epidemiological data were collected on the health status, sexual practices etc. In the prostitute population studied 2.5 % of the non-intravenous drug abusers and 20 % of the intravenous drug abusers were positive for HIV. However, infection with HTLV-I/II could not be demonstrated in any of them.

It is known that ovary and spleen are innervated extensively by afferent and efferent noradrenergic sympathetic nerve fibers from the celiac ganglion. Furthermore, immune cells located in the ovary influence the ovarian physiology. However, the peripheral interaction between the immune and neuroendocrine system is poorly understood. This work was undertaken to study the effect of superior ovarian nerve (SON) transection, in adult rats, on the number of splenocyte β-adrenergic receptors and their possible relation to ovarian steroidogenesis, measuring the effect of secretions of those splenocytes on progesterone and estradiol release from the ovary. Seven days after SON transection, the splenocytes were isolated and then cultured for 48 h. Their number of β-adrenergic receptors, measured using [125I]-cyanopindolol as ligand, increased, and their culture media, used to stimulate ovaries from 60-day-old intact (neither SON-transected nor sham-operated) rats in vitro on diestrous day 2 showed a decrease in progesterone release and an increase in estradiol release in relation to splenocyte culture media of control rats (sham-operated; p < 0.001, respectively). The effects of in vivo SON transection were simulated by an in vitro system modulating the splenocyte β-adrenergic receptor number. The splenocytes from SON-transectioned rats were preincubated with and without norepinephrine (NE) 10–6 M for 48 h, a low and high number of β-adrenergic receptors respectively, and then were stimulated with NE 10–6 M for 24 h. After that, the culture medium from splenocytes with a low number of β-adrenergic receptors induced progesterone release from the ovaries of intact rats (p < 0.001), but produced no change in estradiol release. The data suggest that splenocyte secretions, which participate in the ovarian steroidogenic response, particularly in progesterone release, might be controlled by adrenergic influences since the number of splenocyte β-adrenergic receptors changes through SON-celiac ganglion-noradrenergic postganglionic innervation of the spleen. In estradiol release, probably other neurotransmitters than norepinephrine (NE) are involved when the SON is sectioned. In this paper we also show functional evidence for modulation of immune function by the sympathetic nervous system and its principal neurotransmitter, NE.

As there is insufficient data available on the spread of hepatitis C virus (HCV) infection by sexual transmission, the prevalence of anti-HCV was determined in several sexually promiscuous groups (78 female prostitutes, 55 clients of prostitutes and 146 homosexual men) who denied intravenous drug abuse. The overall HCV infection rate was much higher in the sexually promiscuous groups (8.97 %, 16.36 % and 5.48 % respectively) than in voluntary blood donors (0.48 %), suggesting that HCV infection can be transmitted by sexual intercourse. Consequently, promiscuity may be a risk factor for this infection.

The present study investigates the acute consequences of central adrenergic stimulation on the release of steroids from the ovary. The influence of the superior ovarian nerve (SON) and the relationship between the neural effect and peripheral LH levels were also examined. The intracerebroventricular (i.c.v.) injection of 5 microg epinephrine in SON-intact rats on day 1 of dioestrus (D1) increased progesterone levels in ovarian vein blood from 7 to 21 min after injection but the same injection in SON-intact rats on day 2 of dioestrus (D2) decreased progesterone levels in ovarian vein blood from 1 to 25 min. A smaller dose (0.5 microg) of epinephrine injected i.c.v. in SON-intact rats produced a decrease in progesterone levels in ovarian vein blood of shorter duration. In SON-transected (SONt) animals, 0.5 microg epinephrine i.c.v. caused a smaller decrease in progesterone levels compared with SON-intact rats (P

Two hundred and ninety-four non-transfused prostitutes from Spain, who denied intravenous drug abuse, were tested for hepatitis C virus (HCV) antibodies. Seventeen (5.78 %) of them were seropositive. Both in univariate and correspondence analyses, serological results for hepatitis C were associated with the HIV-1 serostatus (p

The prevalence of silent HIV-1 infections in subjects at risk of acquiring HIV infection by heterosexual transmission was assessed using a nested polymerase chain reaction (PCR) assay. The two groups of risk subjects consisted of 92 female prostitutes and 43 heterosexual partners of infected individuals. Appropriate positive and negative control persons were included in the study. Serum samples were also tested for antibody to HIV-1 by an enzyme immunoassay (EIA) and positive results confirmed by Western blot. PCR results in the two risk groups and the positive and negative controls were in full agreement with serological results. It is concluded that silent infection with HIV-1 is infrequent in persons at risk for heterosexual transmission.

Hyperprolactinemia (HP) induced in female rats by dopaminergic receptor blockers enhanced ovarianestradiol(E3) releasein responseto human chorionic gonadotropin (hCG) or human follicle-stimulating hormone (hFSH) in vitro. Uterine weight and ovarian aromatase activity were also increased. In contrast, ovarian androgen (A) release in response to hCG was reduced. Injectionsof ovineprolactin(oPrI)alsoenhanced E3 responseto hCG in vitro. The increasedE2 response was notdue to a directeffectof PrIon ovarianaromataseactivity since administration of oPrI to hypophysectomized rats failed to enhance the formation of E3 from testosterone (T) in vitro,and inhibitedtheincreaseintheenzyme activity inducedby FSH. Adrenalectomy(ADRX) of intact rats, which did not affect mean serum gonadotropinlevels, bluntedtheeffectof HP on the E2 response to hCG. The suppression was partially reversed by corticosterone (B). Serum progesterone(P)and T were similarin controlsand HP-ovariectomized ratswith intact adrenals.Likewise,serum androstenedione (�4) and dehydroepiandrosterone(DHA) were not altered in intact, HP rats as compared with controls. Thus, an increase in adrenal secretion of these steroids does not appear to mediate the effect of Pri on ovarian E3 responseto gonadotropins. Ovariesof HP ratsshowed more large follicles than controls. In contrast, ovaries of HP-ADRX rats had a decreased number of large follicles. It is suggested that: a) in intact prepubertal rats PrI increases the E2 responseof the ovary to gonadotropins by facilitating follicular development rather than by a direct actionon aromataseactivity, and b) when follicular developmentis stimulated by FSH, an inhibitory effect of PrI on aromatase activity becomes apparent. The effect of PrI on the E2 response of the ovary to gonadotropins is not mediated by the adrenal cortex. Rather, it appears that while PrI facilitates the development of large, E2-producing follicles by promoting the growth of small- and medium-sized follicles, an adrenal component influences follicular growth at a step subsequent to Prl.

In the rat, the main source of adrenergic fibers innervating nonvascular ovarian tissue is the superior ovarian nerve (SON). To determine the influence of the SON on prepubertal ovarian function and, hence, on the time of puberty, several experiments were conducted. Transection of the SON in early juvenile rats (day 24) led to more than 60% depletion in ovarian norepinephrine (NE) content, but affected neither the timing of vaginal opening nor that of first ovulation. SON section, however, produced a 2-fold increase in beta-adrenergic receptor density, with no change in affinity, suggesting that hypersensitivity by denervation had developed. Removal of all adrenergic influences from ovarian granulosa cells by culturing the isolated cells in vitro produced a striking increase in beta-adrenergic receptor number, which was maximal at 72 h. In spite of this elevated receptor content, the beta-adrenergic agonist Zinterol did not induce progesterone release unless the cells were previously exposed for 48 h to a physiological concentration of FSH. Both NE and epinephrine down-regulated the receptors in a dose-related manner, without affecting receptor affinity. To test the hypothesis that denervation of the ovary results in greater steroidal sensitivity to adrenergic stimulation, a model was constructed in which cells were simultaneously exposed to FSH and NE or epinephrine (innervated cells) or exposed only to FSH (denervated cells) for 48 h. The response to adrenergic stimulation was subsequently measured by incubating the cells with Zinterol for 24 h. Zinterol was highly effective in eliciting progesterone release from the denervated cells, but failed to do so in innervated cells. Additionally, the agonist increased cAMP in the former cells, but it was ineffective in the latter, indicating that prior exposure to catecholamines desensitized the adenyl cyclase enzyme coupled to beta-adrenergic receptor sites. The results suggest that during prepubertal maturation, noradrenergic fibers reaching the ovary via the SON may modify ovarian steridogenic sensitivity to catecholamines, by regulating the number of functional beta-adrenergic receptors in the gland. They also indicate that the apparent failure of SON denervation to alter the maturation of ovarian steroidogenesis and the time of puberty may be due to development of compensatory hypersensitivity after the denervation.

The advent of female puberty represents the culmination of a diversity of developmental processes which affect all components of the reproductive axis. Development of neuroendocrine reproductive functions proceeds in a harmonious and interrelated manner. No unique 'trigger' of puberty can be discerned, but rather puberty represents the climax of a cascade of events, finely interconnected throughout the continuum of sexual maturation. A resetting of the hypothalamic 'gonadostat' to steroid negative feedback appears to be a phenomenon associated with puberty, but not its cause. Although the central nervous system plays a pivotal role in the development of the ovary, it is the acquisition of ovarian ovulatory capacity which finally determines the timing of the first preovulatory surge of gonadotropins. In contrast to primates, development of the central component of estradiol positive feedback is an early event in the female rat. However, in most species--including the rat--amplification (or initiation) of a particular, synchronous pattern of LHRH release appears essential for the initiation of puberty. The mechanisms underlying this functional change of the LHRH secreting system are not clearly understood. In the rat, ovarian development proceeds under the influence of gonadotropins, and the somatomammotropins PRL and GH. More intriguingly, evidence is now emerging that the central nervous system may convey direct information to the ovary via the ovarian nerves, thus providing a hormone-independent fine tuning for its control. Upon reaching adequate development, the ovary through its secretory products, acts on an already competent hypothalamic-pituitary axis to activate the central component of estradiol positive feedback.

The rat pars tuberalis was studied with conventional electron microscopy and immunocytochemistry for the demonstration of luteinizing hormone (LH). The LH-secreting cells were preferentially located in two regions of the pars tuberalis (PT), namely, that surrounding the neural stalk and that occupying the tuberoinfundibular sulci. Dialyzed extracts of PT prepared after removal of the pituitary stalk, had the capacity to induce ovulation in chlorpromazine-blocked rats in proestrus. In radioimmunoassays carried out under the same conditions, the PT extracts yielded displacement curves parallel to those of standard LH. The immunoreactive LH content of the female PT was determined at three phases of the estrous cycle: diestrus, afternoon of proestrus and estrus. The lowest values were found in rats sacrificed in the evening of proestrus (18.00 h), and they were about one sixth of the peak values found at estrus. 1 month after inducing lesions in the preoptic area of female rats, the LH content of the PT was four times higher than the highest values found during the estrous cycle. The results suggest that the rat PT does secret LH, and that this secretory activity fluctuates with the estrous cycle, but in a manner that differs from that reported for LH secretion of the pars distalis.

To examine the influence that the vagus nerve may have on ovarian development and on the timing of puberty, both anterior and posterior vagi were sectioned at the abdominal level in 24-day-old female

We have previously shown that, in the rat, ovarian beta-adrenergic receptor content varies during the time of puberty, with values first increasing and then decreasing abruptly on the afternoon of the first proestrus, i.e., at the time of the preovulatory surge of gonadotropins and prolactin (Prl). In the present study, experiments have been conducted to determine: 1) if hormones other than follicle stimulating hormone (FSH) that are known to be involved in regulating prepubertal ovarian function can mimic the facilitatory effect of FSH on progesterone (P) response of granulosa cells to beta-adrenergic stimulation; 2) if beta-adrenergic receptor content of granulosa cells is under hormonal regulation; and 3) whether the facilitatory effect of hormones on the P response to beta-adrenergic stimulation is due to an increased cyclic AMP response to receptor activation. A 48-h in vitro preexposure of granulosa cells from juvenile, 29-day-old ovaries to the pituitary hormones Prl, luteinizing hormone (LH), or FSH showed that only the latter was able to facilitate the subsequent P response to Zinterol, a beta2-adrenergic agonist. Follicle-stimulating hormone also increased basal P release. Of the two nonpituitary hormones examined, the luteinizing hormone-releasing hormone (LHRH) agonist D-(Ala6,Pro9)-LHRH-ethylamide (LHRH-A) failed to affect P responsiveness, whereas corticosterone enhanced both basal P release and P response to Zinterol. This effect was less pronounced than that of FSH. Luteinizing hormone, Prl and corticosterone decreased beta-adrenergic receptor content to different extents, with corticosterone being the most effective and LH the least (50% and 15% decrease, respectively); LHRH-A was ineffective.(ABSTRACT TRUNCATED AT 250 WORDS)

In an attempt to study the involvement of an adrenergic mechanism in the control of ovarian function at puberty, the presence of β-adrenergic receptors wets demonstrated, and the receptors were characterized in pre- and peripubertal ovaries. Using [125I]iodohydroxybenzylpindolol (IHBP) as a ligand, a single class of high affinity, low capacity β-adrenergic sites was found in both granulosa cells and residual ovary. Analysis of Scatchard plots derived from saturation curves or kinetic analysis of association curves provided identical equilibrium dissociation constant (Kd) values for the ligand-receptor binding. Dissociation experiments showed that the ligand binding to the receptor had the presence of both loosely bound, easily displaceable and tight, nondisplaceable components. IHBP binding was inhibited by L-propranolol [inhibition constant (Ki) = 10.5 nM], isoproterenol (Ki = 696 nM), and zinterol, a β2-adrenergic agonist (Ki = 187 nM). Metoprolol, a β1-antagonist, exhibited a much higher Ki (5.3 μM). C...

The ovary and adenohypophysis of the rat contain beta-adrenergic receptors and respond to beta-adrenergic stimulation with hormone release. To determine the importance of the adrenal medulla as a source of adrenergic influences regulating prepubertal ovarian and pituitary function, a technique was developed to remove most of the adrenal medulla without compromising adrenocortical function. Medullectomy (MED) of 24-day-old female rats depressed both spontaneous diurnal changes in plasma epinephrine (EPI), and the EPI and norepinephrine (NE) response to decapitation, without affecting corticosterone (B) levels. Vaginal opening and first ovulation were delayed in MED rats. Serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels were normal in MED rats, but those of growth hormone (GH) and prolactin (Prl) were depressed. MED reduced the ovarian weight response to pregnant mare's serum gonadotropin (PMSG) and the ovarian steroidal response to human chorionic gonadotropin (hCG) in vitro, but it did not affect ovarian beta-adrenergic receptors. Cultured granulosa cells, harvested from juvenile ovaries and primed in vitro with FSH, responded to nanomolar concentrations of EPI with progesterone (P) secretion. EPI also augmented hCG- and FSH-induced P secretion. The EPI effect was reproduced by Zinterol, a beta 2-adrenergic agonist and was prevented by propranolol, a beta-adrenergic antagonist. Blockade of alpha-adrenergic receptors with phentolamine was ineffective. It is suggested that EPI of adrenomedullary origin supports female prepubertal development by a) stimulating ovarian P secretion, b) favoring Prl and GH release and c) amplifying the stimulatory effect of low gonadotropin levels on ovarian steroidogenesis. The effects of EPI on ovarian function appear to be mediated by beta-adrenergic receptors of the beta 2 type.

During the days preceding the first ovulation the ovary of the rat exhibits a remarkable increase in estradiol (E2) and progesterone (P) release in response to gonadotropins. No such increase is observed in the case of androgens (A, testosterone + dihydrotestosterone). The present experiments were undertaken to examine the possibility of reproducing these developmental events by stimulating the ovary with a gonadotropin that has substantial FSH-like activity. In vivo administration of pregnant mare serum gonadotropin (PMSG) to juvenile 29-day-old rats greatly increased the in vitro E2 and A response to human chorionic gonadotropin (hCG) measured 2 days later in the morning. The magnitude of the A response was significantly larger than that of ovaries from juvenile animals or rats in first proestrus. The E2 response was much greater than that of juvenile ovaries but similar to that of ovaries from late proestrous rats. In contrast, the P response to hCG was not enhanced by PMSG. In fact the response was similar to that of juvenile ovaries and markedly less than that of first proestrous rats. This decreased P response was not due to a greater conversion of P to its less active metabolite 20 alpha-hydroxy-4-pregnen-3-one (20 alpha-OH-P). The results suggest that PMSG enhances the E2 and A response of immature ovaries to hCG at the expense of that of P. Treatment of immature rats with PMSG may represent a useful model to study E2 release from preovulatory ovaries, but it cannot be used to reproduce in its entirety the developmental changes in steroidal response to gonadotropins associated with normal puberty.

A system of intercellular channels is described in the pars tuberalis (PT) of the female rat. These spaces are lined by all types of cells found in the PT and are not sealed off by tight junctions. Ventrally and dorsally, the intercellular spaces open toward the basement membranes separating the PT from (i) the subarachnoid space, and (ii) the perivascular space of the portal capillaries, respectively. These intercellular channels differ from the follicles, which are also found in the PT, being lined by a particular type of cell. In a second group of female rats an epoxy mixture was injected into the third ventricle; 10 min thereafter horseradish peroxidase was infused into the cisterna magna. After processing the brain for the demonstration of exogenous peroxidase, it was found that the tracer had reached the subarachnoid space adjacent to the hypothalamus and entered into all ventricular cavities with the exception of the infundibular recess. Under these experimental conditions it was found that the tracer fills all intercullular channels of the PT, thus indicating that there is no barrier between the subarachnoid space and the PT. It is suggested that the subarachnoid space should be regarded as a probable route for the transport of trophic factor(s) and/or secretory product(s) of the PT.

Intact female rats received transplants of two hypophysial partes distales under the kidney capsule. The plasma levels of prolactin were determined, and the ultrastructure of the grafted gland was studied 15, 45 and 90 days after the operation. Although prolactin levels in the three experimental groups were significantly higher than those in control rats, a decrease in prolactin level was detected in the 45-day samples. Parallel ultrastructural changes suggest that between the 45th and 90th postoperative day a process is initiated leading to hyperplasia and hypertrophy of prolactotrophs.

Two hypophysial partes distales were grafted under the kidney capsule of intact female rats. The plasma prolactin levels 15, 45 and 90 days after the operation were determined. At the same postoperative intervals the grafted glands of some of the operated rats were processed for conventional light and electron microscopy and for the demonstration of prolactin, FSH and LH according to the unlabelled immunoperoxidase procedure. The ultrastructural characteristics of the transplanted secretory cells and the amount and distribution of the immunoreactive material within their cytoplasm were used to evaluate approximately the secretory activity of these cells. Although levels of prolactin in the three experimental groups were significantly higher than those in control rats, a decrease in prolactin level was detected in 71% of the samples taken 45 days after operation. At day 15 the graft was completely surrounded by lymphoid cells whereas at day 45 these cells had invaded the whole graft. In the group sampled at day 90 the graft was free of lymphoid cells. When traced immunocytochemically the three types of cells followed different patterns of evolution after transplantation. Most prolactotrophs were hypertrophied in all groups but, in addition, they underwent a process leading to hyperplasia some time between days 45 and 90 after operation. Syncytial formations which probably correspond to multinucleated prolactotrophs were present only in the group sampled at day 90. The number of LH and FSH cells had decreased in the group at day 45 and by day 90 the former remained scarce but immunoreactive FSH cells were no longer found. At the ultrastructural level clear signs of involution of gonadotrophs and degradation by macrophages were seen in the graft 45 days after operation. The relation between the morphology and hormone content of the graft and hormone content of the plasma is discussed, together with several questions raised by the results. Pituitary transplantation can be used as an experimental model only if the time-dependent changes described here are taken into account.