Effect of Bifidobacterium infantis NLS super strain in symptomatic coeliac disease patients on long-term gluten-free diet - an exploratory study.

Bifidobacterium infantis NLS super strain (B. infantis NLS-SS) was previously shown to alleviate gastrointestinal symptoms in newly diagnosed coeliac disease (CD) patients consuming gluten. A high proportion of patients following a gluten-free diet experiences symptoms despite dietary compliance. The role of B. infantis in persistently symptomatic CD patients has not been explored. The aim of the study was to evaluate the effect of B. infantis NLS-SS on persistent gastrointestinal symptoms in patients with CD following a long-term GFD. We conducted a randomised, cross-over, double-blind, placebo-controlled trial in symptomatic adult CD patients on a GFD for at least two years. After one-week run-in, patients were randomised to B. infantis NLS-SS or placebo for 3 weeks with cross-over after a 2-week wash-out period. We estimated changes (Δ) in celiac symptom index (CSI) before and after treatment. Stool samples were collected for faecal microbiota analysis (16S rRNA sequencing). Gluten immunogenic peptide (GIP) excretion in stool and urine samples was measured at each study period. Eighteen patients were enrolled; six patients were excluded due violations in protocol. For patients with the highest clinical burden, CD symptoms were lower in probiotic than in placebo treatment (P=0.046). B. infantis and placebo treated groups had different microbiota profiles as assessed by beta diversity clustering. In probiotic treated groups, we observed an increase in abundance of B. infantis. Treatment with B. infantis was associated with decreased abundance of Ruminococcus sp. and Bifidobacterium adolescentis. GIP excretion in stools and urine was similar at each treatment period. There were no differences in adverse effects between the two groups. B. infantis NLS-SS improves specific CD symptoms in a subset of highly symptomatic treated patients (GFD). This is associated with a shift in stool microbiota profile. Larger studies are needed to confirm these findings. ClinicalTrials.gov: NCT03271138.

Fertilization and connectivity in the Garrucha Canyon (SE-Spain) implications for Marine Spatial Planning.

Marine Spatial Planning is usually based on benthic georeferenced information or GPS tracked human activities, whereas the pelagic ecosystem is often ignored because of scarce and limited surface information. However, the 3-D pelagic ecosystem plays a key role connecting all the other ecosystems by physical (currents) and biological (migration) processes. According to remote sensing the Garrucha Canyon is oligotrophic, but 3-D sampling reveals subsurface upwelling, and converts it into the richest area around the Cape of Gata. Vertical connectivity by means of zooplankton migration, measured at two sampling stations, is 40 and 220 times faster than microphytoplankton settling and vertical water velocities respectively. Thus coupled physical-biological connectivity models are necessary to estimate the ecosystem connection and the fate of carbon, but also other substances (e.g. radioactivity), that might accumulate throughout the food-web. This is especially important in the Garrucha Canyon and the Coastal Areas Management Programme Levante de Almería where natural heritage and extractive fishery are important for the local economy.

Analysis and characterization of cultivable extremophilic hydrolytic bacterial community in heavy-metal-contaminated soils from the Atacama Desert and their biotechnological potentials.

AIMS: To isolate and characterize the cultivable community of hydrolase producers (amylase, protease, lipase, DNase, xylanase and pullulanase) inhabiting heavy-metal-contaminated soils in extreme conditions from the Atacama Desert. METHODS AND RESULTS: A total of 25 bacterial strains showing hydrolytic activities have been selected including halotolerants, extremely halotolerants and moderate halophiles. Most hydrolase producers were assigned to the family B acillaceae, belonging to the genera Bacillus (nine strains), Halobacillus (seven strains) and Thalassobacillus (five strains) and four isolates were related to members of the families Pseudomonadaceae, Halomonadaceae and Staphylococcaceae. The selected strains were then characterized for their tolerance pattern to six heavy metals, measured as minimal inhibitory concentrations (MICs). CONCLUSIONS: The diversity found in the cultivable bacterial community analysed is more limited than that detected in other ecological studies owing to the restrictive conditions used in the screening. The dominant bacteria were Firmicutes and particularly, species related to the genus Bacillus. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is focused on the characterization of extremophilic hydrolytic bacteria, providing candidates as a source of novel enzymes with biotechnological applications.

Clinical utility of counting intraepithelial lymphocytes in celiac disease intestinal mucosa / Utilidad clínica del recuento de linfocitos intraepiteliales en la mucosa intestinal para el diagnóstico de enfermedad celíaca

BACKGROUND/AIMS: Our aims were to establish the clinical utility of assessing the intraepithelial lymphocyte (IEL) density in intestinal biopsies from a large series of individuals and to determine the best threshold discriminating celiac disease (CD) patients and controls in two populations with different pre-test prevalence. METHODS: We prospectively performed intestinal biopsy and CD-related serology in 349 subjects undergoing upper GI endoscopy. While 116 had symptoms suggestive of a small bowel disorder (high prevalence), 233 individuals were randomly selected from patients referred to endoscopy because upper GIsymptoms (low prevalence). Diagnosis of CD was based on the concordance of classical histological features and a positive CD serology. RESULTS: While 58 patients had a newly diagnosed CD (52 in the high and 6 in the low prevalence groups), 291 subjects did not meet diagnostic criteria of the disorder. Patients had a highly significant greater IEL density than controls (p < 0.00001). Based on the ROC curve, a count of 22.8 IEL/100 epithelial cells had the highest performance for diagnosing CD in the overall population and for subjects in the high pre-test probability subgroup and 22.5% was ,he best cut-off for those diagnosed in the low risk population (area under the curves: 0.979, 0.979 and 0.993, respectively). An abnormal CD serology confirmed the diagnosis of CD in all the four patients with counts below 22.8%. CONCLUSIONS: Our study confirms that an IEL density of 22.8% is an adequate threshold to discriminate CD patients and controls in individuals irrespective of the prevalence of the disorder.

Introducción: El recuento elevado de linfocitos intraepiteliales (LIEs) es un rasgo destacado aunque inespecífico de la enteropatía de la enfermedad celíaca (EC). Un recuento mayor a 40 LIEs/100 células epiteliales ha sido considerado por mucho tiempo esencial para el diagnóstico. Sin embargo, estudios recientes con escaso número de muestras han cuestionado este valor de corte. Objetivos: Determinar el rango normal de LIEs en biopsias intestinales y establecer su capacidad diagnóstica de EC en dos poblaciones con diferente prevalencia. Métodos: Realizamos prospectivamente biopsias de duodeno distal y serología para EC en 349 pacientesconsecutivos a quienes se les realizó una videoendoscopia digestiva alta. El grupo A consistió en 116 pacientes derivados a biopsia intestinal por síntomas sugestivos de malabsorción (considerados de alta prevalencia de EC) y el grupo B consistió en 233 pacientes randomizados entre quienes fueron derivados a endoscopía alta por síntomas gastrointestinales no sugestivos de EC (baja prevalencia de EC). El diagnóstico de EC se basó en criterios histológicos clásicos y serología positiva. Resultados: Cincuenta y ocho pacientes tuvieron EC (52 en el grupo de alto riesgo y 6 en el de baja prevalencia) y 291 individuos no tuvieron criterios de la enfermedad. Los pacientes tuvieron una densidad de LIEs significativamente mayor que los controles (p<0.00001). Basado en las curvas ROC, el conteo de 22.8 LIEs/100 células epiteliales tuvo la mejor sensibilidad y especificidad para el diagnóstico de EC en la población general y entre los sujetos con alta probabilidad y 22.5% fue el mejor valor de corte para la población de bajo riesgo (áreas bajo las curvas: 0.979, 0.979 y 0.993, respectivamente). Todos aquellos pacientes celíacos con recuento de LIEs por debajo de 22% (n=4), tuvieron serología positiva para EC. El clásico valor de 40% tuvo una sensibilidad del 55%. Conclusiones: Nuestro estudio confirma que una...

The IL-1 gene family and bone involvement in celiac disease.

Celiac disease (CD) is associated with decreased bone mineral mass. Its pathogenesis is multifactorial since both systemic and local mechanisms may play a role. Our objective was to determine whether single-nucleotide polymorphisms in genes encoding members of the interleukin-1 family are associated with bone damage measured by densitometry in a series of 71 adult CD patients assessed at diagnosis. When compared with non-carrier CD patients, carriers of allele T of the interleukin-1beta gene (IL1B-511T) had a significantly lower bone mass at the total skeleton level (p = 0.0484) and a greater prevalence of osteopenia/osteoporosis (p = 0.0102). To our knowledge, this is the first evidence on the association between a genetic predisposition and low bone mass in CD patients. This finding supports the postulated inflammation-associated bone loss pathogenesis as one of the causes of bone weakness in CD.

Significance of smooth muscle/anti-actin autoantibodies in celiac disease / Significado de los anticuerpos anti-músculo liso/anti-actina en la enfermedad celíaca

BACKGROUND/AIM: Smooth muscle antibody (SMA) specific for the protein actin, a major component of the cytoskeleton of epithelial cells, is one of the most prevalent non-organ specific autoantibodies in the serum of celiac disease (CD) patients. Our aim was to explore the clinical relevance of the presence of IgA type anti-actin antibody (AAA) and SMA in a series of patients with CD. METHODS: We evaluated frozen serum samples collected at diagnosis from 92 adult patients with CD and 52 control individuals in whom CD was excluded. Patients were re-evaluated a median time of 5 yr after treatment. IgA type AAA was detected using a modified commercial ELISA assay and IgA SMA was detected using indirect immunofluorescence on primate esophagus substrate. RESULTS: At diagnosis, samples from CD patients had significantly higher AAA values than controls (p<0.00001). While all active CD patients had serum AAA values over the cut-off for healthy controls, we observed a very significant reduction of these antibodies after treatment (p>0.0001). AAA had a highly significant correlation with both, tissue, transglutaminase (r=0.62) and antigliadin (r=0.60, p<0.00001) antibodies as well as the severity of the intestinal injury (p<0.05). SMA was detected in sera of 35 consecutive CD patients. At diagnosis, SMA positive patients had significantly higher values of AAA (p<0.0002), increased number of autoimmune disorders (p<0.04), delayed menarche (p<0.04), lower hemoglobin levels (p<0.01), increased fecal a-I antitrypsin clearance (p<0.01) and more severe diarrhea (p<0.06). We also detected a trend to more severe complications at follow-up (p=0.059). CONCLUSIONS: Based on our findings we suggest that the presence of increased IgA AAA serum levels is a highly sensitive marker of the disturbed architecture of intestinal epithelial cells of CD patients with a potential relevance to diagnosis and follow-up. The presence of SMA seems to define a distinct subset of CD patients with ...

Introduccion/objetivo: El anticuerpo anti-musculo liso (SMA) dirigido contra la proteína actina, un componente mayor del citoesqueleto de las células epiteliales, es el anticuerpo no-órgano específico más prevalente en enfermedad celíaca (EC). Nuestro objetivo fue explorar la importancia clínica de los anticuerpos anti-actina (AAA) y SMA en una serie de pacientes con EC. Métodos: Evaluamos 92 muestras serológicas de pacientes celíacos adultos recolectadas al momento del diagnóstico y la de 52 individuos controles no celíacos. Los pacientes fueron re-evaluados luego de un tiempo medio de 5 años en tratamiento. Evaluamos AAA tipoIgA mediante ELISA empleando un equipo commercial modificado y SMA IgA por inmunofluorescencia indirecta sobre sustrato de esófago de mono. Resultados: Al momento del diagnóstico, los pacientes celíacos tuvieron valores de AAA significativamente más elevados que los controles (p<0.00001). Todos los pacientes con EC activa presentaron niveles de AAA por encima del valor de corte determinado para el grupo control sano y se evidenció una reducción significativa de los nivelesluego del tratamiento (p>0.0001). Los AAA presentaron una correlación significativa con los anticuerpos anti-transglutaminasa tisular (r=0.62) y anti-gliadina (r=0.60) (p<0.00001), de igual modo que con la severidad del daño intestinal (p<0.05). Al momento del diagnóstico, se detectó SMA en el suero de 35 pacientes no controles. Los pacientes SMA positivos tuvieron valores significativamente mayores de AAA (p<0.002), un incremento del número de enfermedades autoinmunes asociadas (p<0.04), menarca tardía (p<0.04), niveles bajos de hemoglobina (p<0.01), incremento del clearance de a-1 antitripsina fecal (p<0.01) y mayor severidad de la diarrea (p<0.06).En ellos se evidenció una tendencia al desarrollo de complicaciones más severas durante el seguimiento (p=0.059). Conclusiones: Sugerimos que la presencia de un valor sérico aumentado de AAA tipo IgA...

Antibodies to human recombinant tissue transglutaminase may detect coeliac disease patients undiagnosed by endomysial antibodies.

BACKGROUND: The screening and diagnosis of coeliac disease have been simplified by the advent of new serological tools. AIM: To assess the clinical utility of a newly developed kit for antibodies to human recombinant tissue transglutaminase (hu-anti-tTG) in a large population of patients undergoing intestinal biopsy for suspected intestinal disorders. METHODS: We evaluated 426 serum samples from consecutive adult patients (250 from untreated coeliac disease patients and 176 from individuals in whom a diagnosis of coeliac disease had been excluded), obtained at the time of intestinal biopsy. Samples were tested for immunoglobulin A (IgA) hu-anti-tTG by enzyme-linked immunoabsorbent assay, IgA endomysial antibodies (EmA) by indirect immunofluorescence and IgA and IgG antigliadin antibodies by enzyme-linked immunoabsorbent assay. A sub-group of samples was also assessed for a guinea-pig-based anti-tissue transglutaminase. RESULTS: According to the cut-off for hu-anti-tTG, the sensitivity, specificity and positive and negative predictive values were 91%, 96%, 97% and 87%, respectively. Simultaneous determination of EmA showed values of 86%, 100%, 100% and 83% for the same parameters. Although 19 coeliac disease patients (7.6%) were negative for EmA and hu-anti-tTG, both tests rendered superior statistical values to antigliadin antibody tests. At diagnosis, IgA deficiency was detected in 11 patients, but both assays were able to detect samples with mild to moderate deficiency. The comparison of hu-anti-tTG with EmA showed excellent concordance between the tests (kappa statistic, 0.85). Discordance was observed in 20 samples from coeliac disease patients (8%) and in nine samples from controls (5%). Fifteen samples had an EmA-negative but hu-anti-tTG-positive serology, and five showed the converse pattern. Comparison of human recombinant and guinea-pig tests showed concordant results in 96% of cases. CONCLUSIONS: The quantitative determination of hu-anti-tTG type IgA using a commercial enzyme-linked immunoabsorbent assay kit was highly sensitive and specific for the detection of coeliac disease. Our results in a large population of patients with a clinical condition suggestive of the disorder demonstrated that the test can be used to detect a substantial number of patients otherwise unrecognized by IgA EmA.

[In-hospital Radio-Pharmacy Unit: creation and development]. / Unidad de Radiofarmacia Hospitalaria: creación y desarrollo.

Regarding the creation of an In-Hospital Radio-Pharmacy Unit within Clínica Puerta de Hierro University Hospital, as well as the compliance with Real Decreto 479/1993 on the regulation of radio-drugs for human use, the steps to be taken to bring it service are laid out and discussed. An overall review of the differential characteristics of radio-drugs is undertaken. The adequate distribution of spaces and the Unit's functioning regulations are presented, and the responsibilities of the person in charge of the ionisation chamber are clearly placed on record. The professional responsible of the Unit should define performance and quality control protocols for each test. The responsibility for coordinating the Unit's operation is assigned to the Heads of Nuclear Medicine and Pharmacy Departments, within the Pharmacy and Therapeutics Committee and the Nuclear Medicine Quality Committee.

[Frecuency of adverse events during the hygiene of the critical care patient]. / Frecuencia de eventos adversos durante el aseo del paciente crítico.

The hygiene of the patient in critical condition is a common nursing technique in the intensive care unit, which does not mean that doing it is exempt of risk for the patient's state. We carry out a study to measure the frequency of the appearance of certain adverse events during the hygiene care and their clinical repercussion.Hygiene of the critical patients was monitored, measuring the appearance of certain events at the time of hygiene and until one hour after to assess if the complications were at the moment or had a greater repercussion on the state of the patient.During the study period, some adverse event appeared in 48% (CI 95%: 43-52) of the hygiene performed while none appeared in 52% (CI 95%: 48-56) of it. The events that appeared most frequently were: desaturation in 18% (CI 95%: 15-21) of the hygiene performed, the deadaptation of the mechanical ventilation in 11% (CI 95%: 9-14), arterial hypertension in 21% (CI 95%: 18-25) and arterial hypotension in 11% (CI 95%: 9-14). The intracranial hypertension appeared in 42% (CI 95%: 26-61) of the hygiene performed to patients who were carriers of intraventricular catheter, 9% (CI 95%: 2-25) continued with elevated values 1 hour after concluding the hygiene. The rest of the events monitored presented a lower frequency, although the appearance of one episode of cardiorespiratory arrest and two of auricular fibrillation with rapid ventricular response, one of which required cardioversion, stand out. We conclude that it is an essential job of the nursing staff to correctly assess the risks that the performance of hygiene means for the critical patient, so that the technique should be applied rationally and under strict monitoring and control.

[Analysis of costs and cost per diagnostic group of critically burned patients in the Spanish public health care system]. / Análisis de la financiación y del coste por proceso de los enfermos quemados críticos en el sistema público de salud español.

The financing of the National Institute of Health (INSALUD) of Spain will soon be based on Diagnosis-Related Groups (DRGs). Knowledge of the real cost of different DRGs is fundamental to ensure adequate financing and to establish criteria for comparisons between centers. Our public health system has no data on the real cost of critically burned patients and their DRGs. This retrospective descriptive study was carried out in a Major Burns Unit (MBU) and included all patients admitted between January and December 1996. Real total cost of the care of critical burned patients, cost per patient, and cost per DRG related with critical burn patients were calculated for the study period. Financing by Weighed Care Units (WCU) was compared with real costs. The total cost of the care of critical burn patients was 346,298,872 Spanish pesetas and the cost per patient was 4,439,729 ptas. WCU financing was 322,021,616 ptas and 4,128,482 ptas, respectively. The DRG with the highest total cost was 458 (non-extensive burns with skin grafts, 106,372,016 ptas). The DRG with the highest average cost was 472 (extensive burns with surgical procedure, 5,401,119 ptas). The DRG with the highest cost per stay was 457 (extensive burns without surgical procedure, 404,683 ptas). For the first time in Spain, the cost of DRGs related with critical burn patients is described. This information is necessary for DRG-based allocation of funds and for establishing criteria to compare centers. The real cost of critical burn patients exceeded WCU financing.

Virulence factors and pathogenicity of Vibrio vulnificus strains isolated from seafood.

The virulence factors of Vibrio vulnificus are not yet well understood. So far, many hydrolytic enzymes have been implicated in the pathogenesis of this micro-organism. The present research was carried out in order to study the presence of some of these enzymes in 133 V. vulnificus strains isolated from 45 seafood samples. The results showed that 100% of these strains were positive for the production of lecithinase and lipase (Tween-80), 99.2% for caseinolytic protease, 96.9% for DNase, 65.4% for mucinase and 46.6% for elastase. None of the strains was positive for the production of collagenase and 96% were haemolytic against sheep blood cells. In relation to colony morphology on brain heart infusion (BHI) agar and nutrient agar, 59.4% of strains showed opaque morphology on BHI agar and 57.9% on nutrient agar, 10.5% presented translucent morphology on both agars and 30.1 and 31.6% of strains showed a mixture of opaque and translucent morphology on BHI agar and nutrient agar, respectively. None of the translucent colonies was virulent to mice. Therefore, opacity was a useful marker for potential virulence. Of 45 food samples contaminated with V. vulnificus, 29 (64.4%) presented strains lethal to adult mice.

[Incidence of pneumonia due to mechanical ventilators in relation to external circuit changes in the respirator]. / Incidencia de neumonía asociada a ventilación mecánica en relación al cambio de circuitos externos del respirador.

Pneumonia is one of the infections of highest relevancy in Intensive Care Units, and according to the incidence of pneumonia associated to mechanical ventilation, the frequence of the ventilator external circuits change is still a topic of discussion. We decided to modify the protocol of change in our unit, from doing it every 48 hours to every 7 days. We performed a prospective study in 108 patients attended in a Polyvalent ICU who underwent mechanical ventilation during more than 24 hours. We formed two groups, in Group 1 we changed circuits every 48 hours and in Group 2 the circuits were changed every 7 days, without using bacterian filters in any of the groups. The results obtained in Group 1 were of an accumulated incidence of pneumonia associated to mechanical ventilation of 18% and density of incidence of 21.1 pneumonias per 1000 days of mechanical ventilation. In Group 2 we obtained an accumulated incidence of pneumonia associated to mechanical ventilation of 19% and a density of incidence of 20.5 pneumonias per 1000 days of mechanical ventilation. In the analysis of data there were no significant statistic differences between both groups. The cost of respirator external circuits was diminished in 27% in Group 2. We conclude that the circuits change every 7 days does not produce an increase in the frequence of pneumonia associated to mechanical ventilation, with the expense of the respirator external circuits being remarkably reduced.

Interlaboratory study of the analysis of ampicillin by liquid chromatography.

A liquid chromatographic method for the analysis of ampicillin was examined in a collaborative study involving seven laboratories. The method included an isocratic part, which is used in the assay. The isocratic part is similar to the assay method for ampicillin of the US Pharmacopeia XXIII Revision. When the isocratic part is combined with gradient elution, the method is suitable for purity control. Six samples of ampicillin (anhydrous, trihydrate and sodium salt) with varying purity were analysed. The main component and related substances were determined. An analysis of variance proved the absence of consistent laboratory bias. The laboratory-sample interaction was significant. Estimates of the repeatability and reproducibility of the method, expressed as standard deviations of the result of the determination of ampicillin, were calculated to be about 0.9 and 1.1 respectively.

Evaluation of liquid chromatography methods for the separation of ampicillin and its related substances.

Two laboratories collaborated to examine the selectivity of four isocratic liquid chromatography (LC) methods for the separation of ampicillin and its related substances. The United States Pharmacopeia (USP) assay method gave the best selectivity. Similar selectivity was obtained on C18 columns as well as on C8 and poly(styrene-divinylbenzene) copolymer columns. A resolution test using cefradine was proposed to replace the test with caffeine prescribed by the USP. Based on the USP method, a gradient LC method was developed for the analysis of related substances in ampicillin. This LC method has been proposed for assay and purity control in the ampicillin monographs of the European Pharmacopeia.

Cyclosporine effects on in vitro responsiveness of anterior pituitary hormone release to dopamine and thyrotropin-releasing hormone in young female rats.

Endocrine side effects of the immunosuppressive drug cyclosporine (CyA) include changes in anterior pituitary hormone secretion. The aim of the present study was to examine the effects of CyA on the responsiveness of in situ and ectopic anterior pituitary prolactin (PRL), growth hormone (GH) and luteinizing hormone (LH) release response to dopamine (DA) and thyrotropin-releasing hormone (TRH) treatment in young female rats, and to evaluate the possible PRL participation in these effects. Thirty day old rats were rendered hyperprolactinemic by transplanting an anterior pituitary gland of a littermate donor, under the kidney capsule, and were then injected with CyA or vehicle for 2 or 8 days. Sham-operated rats were used as controls and treated in the same way. CyA treatment prevented the increase in plasma PRL levels which occurred in controls after pituitary grafting. In vitro basal PRL release of in situ pituitaries from either sham-operated and/or pituitary-grafted animals was decreased by CyA treatment at any point studied. Basal in vitro secretion of GH was only decreased in the in situ pituitaries from grafted animals after 2 days of CyA therapy. The presence of an ectopic pituitary lead to an increase in the in vitro basal LH secretion from in situ pituitaries as compared to those from sham-operated rats. Basal LH release rates were not changed by CyA treatment, although the LH release in vitro did increase in the in situ pituitaries from sham-operated animals treated with the drug for 2 days. DA addition to the incubation media decreased the in vitro release of PRL, GH and LH from the in situ pituitaries of sham-operated and pituitary-grafted animals treated with vehicle. In CyA treated animals, DA decreased in vitro PRL release from the in situ pituitaries of animals, independently of the presence or absence of an ectopic pituitary. Reductions of the in vitro GH and LH release release after DA treatment were higher in the in situ pituitaries from grafted animals on day 8 of CyA or vehicle treatment. TRH increased the in vitro release of the three hormones with differential effects related to the length of the treatment with CyA and/or the presence of an ectopic pituitary. In vitro release of PRL and GH by ectopic pituitaries was inhibited by previous treatment with CyA and this effect was decreased proportional to the duration of the treatment with the drug, while LH secretion was not modified. Addition of DA to the incubation media resulted in a marked reduction of in vitro PRL and GH release, but only at day 8 of vehicle treatment on GH release did DA addition to media further decrease the release of both hormones from ectopic pituitaries from animals treated for 2 or 8 days with the drug, whereas LH secretion was not modified. TRH addition to the incubation media of ectopic pituitaries surprisingly reduced PRL and GH secretion on day 8 of CyA treatment or after surgery. The results of these studies suggest that CyA can act directly at the hypophyseal level modifying pituitary responsiveness to external stimuli. CyA seems to exert its main effects on lactotroph activity while its effects on somatotrophs and gonadotrophs are less.

Effects of cyclosporine on ovarian function in sham-operated and pituitary-grafted young female rats.

Effects of cyclosporine (CyA) on ovarian function and the possible role of prolactin in mediating these effects were examined in young female rats. The animals were sham-operated or rendered hyperprolactinemic by transplanting pituitary glands under the renal capsule. Cyclosporine prevented the increase in plasma prolactin levels in grafted rats. However, in sham-operated animals plasma prolactin levels were increased after 8 days of CyA treatment. Plasma levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were reduced 8 days after pituitary grafting and increased by CyA at both Day 2 and Day 8 of treatment. The content of LHRH in the hypothalamus was not affected on Day 2 but was reduced on Day 8 after grafting on CyA therapy. Plasma estradiol levels were increased by CyA in sham-operated rats on Day 2 and 8 of treatment, and in pituitary-grafted rats on Day 8 of therapy. In sham-operated rats, ovarian estradiol content was reduced after 2 and after 8 days of CyA administration. In pituitary-grafted rats, the ovarian estradiol content was suppressed after 8 days, and CyA treatment prevented this effect. Ovarian estradiol release in vitro under basal conditions was greater in ovaries derived from 38-day-old than in those from 32-day-old animals. The ovarian estradiol response to human chorionic gonadotropin (hCG) in vitro was increased 2 days after pituitary transplantation. Administration of CyA for 8 days increased basal and hCG-stimulated estradiol release in both sham-operated and pituitary-grafted animals. The present findings suggest that CyA can alter ovarian function by acting directly at the gonadal level. However, a hypothalamic-hypophyseal site of action cannot be ruled out.

Effects of cyclosporine at the hypothalamic-pituitary axis in pituitary-grafted young female rats.

This work was designed to investigate the effects of cyclosporine on prolactin secretion by an ectopically grafted heterologous pituitary gland, and on the hypothalamic content of norepinephrine, dopamine and serotonin. The administration of cyclosporine prevented the augmentation in plasma prolactin levels which occurred following an ectopic graft of a litter-mate pituitary gland. In contrast, in sham-operated rats, cyclosporine increased prolactin levels on day 8 of treatment. Both pituitary grafting and cyclosporine treatment in sham-operated rats decreased hypothalamic norepinephrine content. In grafted rats, cyclosporine returned hypothalamic norepinephrine to normal. Hypothalamic serotonin content decreased 8 days after pituitary grafting but increased to the values of control animals after cyclosporine administration. Cyclosporine treatment for 2 and 8 days increased serotonin content in sham-operated animals. As expected, the hypothalamic dihydroxphenylacetic acid/dopamine index increased after pituitary grafting and administration of cyclosporine for 8 days resulted in a further increase. Cyclosporine administration for 2 days, however, decreased this index to the values observed in control animals while drug treatment of control rats for 8 days decreased the dihydroxyphenylacetic acid/dopamine index. In vitro release of prolactin from the ectopic gland was markedly decreased in animals treated with cyclosporine for 2 days and this effect was less evident in 8-day treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Thymostimulin effects on the pituitary-ovarian axis in hyperprolactinemic young female rats.

This work was undertaken to study the effects of thymostimulin on the pituitary-ovarian axis of young female rats, and to elucidate possible prolactin-mediated effects. 30-day-old rats implanted with an ectopic pituitary from a littermate male donor under the kidney capsule or sham-operated animals were injected with thymostimulin or vehicle for 2 and 8 days. Chronic thymostimulin treatment to sham-operated rats increased plasma prolactin and estradiol levels, and decreased gonadotropin levels. Thymostimulin administration to pituitary-grafted rats did not further modify the increase in plasma prolactin and the decrease in plasma luteinizing hormone levels induced by the presence of an ectopic pituitary, suggesting a hypothalamic and/or hypophysial site of action of the thymic fraction. An increase in plasma estradiol and progesterone levels was observed with the in vivo thymostimulin treatment, although differential in vitro effects on estradiol responses to human chorionic gonadotropin in the various groups studied were observed. These effects seem to be mediated through the changes in plasma prolactin and gonadotropin concentrations observed. The above-mentioned changes induced by the thymic fraction on plasma estradiol and progesterone levels might counteract prolactin effects on the thymus. These data suggest that thymostimulin treatment of young female rats changes hypophysial and sex hormone secretion, as well as the response of the ovaries to human chorionic gonadotropin, probably modifying the normal process of sexual maturation.

Effects of naloxone on pulsatile luteinizing hormone in experimental hyperprolactinemia.

It is well known that opioids regulate luteinizing hormone (LH) secretion through not fully understood mechanisms. This study describes the effects of naloxone on the episodic release of LH in adult sham-operated and pituitary-grafted female rats. Animals were rendered hyperprolactinemic by transplanting 2 pituitary glands beneath the kidney capsule. Naloxone (2 mg/kg/h) or saline (0.5 ml/h) were administered iv through jugular cannulae and subjects were bled at 7 min intervals for a period of 3 h. As expected, pituitary-grafting was followed by an increase in mean values of prolactin during the bleeding period. Naloxone administration decreased mean serum prolactin levels in sham-operated rats and did not further change them in pituitary-grafted animals. Hyperprolactinemia was associated with increases in mean serum LH levels during the bleeding period and in the absolute amplitude of LH peaks. Naloxone administration increased mean values of LH and the absolute amplitude of LH peaks, and decreased their frequency in sham-operated rats. Neither pituitary grafting nor naloxone administration modified the frequency, duration or relative amplitude of LH peaks. However, naloxone administration reduced the mean half-life of LH in sham-operated rats to a similar extent than did pituitary grafting. Naloxone failed to further change the mean half-life of LH in pituitary-grafted rats. These results suggest that opioids modulate the pulsatile pattern of LH and that these effects are blunted in pituitary-grafted animals.