Hepatic presentations of mitochondrial DNA depletion syndrome in children: A single tertiary liver centre experience.

Mitochondrial DNA depletion syndromes (MDDS) are a heterogeneous group and the hepato-cerebral phenotype is highly variable. A single centre retrospective study of all patients with MDDS presenting between January 2002 and September 2019. In total, 24 (13 male) children were identified: 7 POLG, 7 DGUOK, and 10 MPV17. Median age at presentation was 3 months (0.06-189). Sixteen had acute liver failure (ALF) and eight chronic cholestasis and/or raised transaminases. Four POLG patients developed liver injury after starting sodium valproate; Six DGUOK patients had neonatal ALF (median age 12 days), liver involvement developed at a median age of 2.5 and 11 months with MPV17 and POLG patients, respectively. Eighteen patients showed neurological involvement. Liver histology from 10 patients showed variable degrees of necrosis, steatosis, cholestasis, and fibrosis. Mitochondrial respiratory chain enzymology was abnormal in 5. Seventeen patients died at a median age of 8 months (range, 1-312) after a median time of 5.6 months from presentation: 5/7 POLG at 53 months, 7/7 DGUOK at 8 months and 5/10 MPV17 at 8 months. Three patients with MPV17 mutations received liver transplant (LT) at a median age of 24 months (range 5-132): all alive at 19, 18 and 3 years post-LT. Mutations in DGUOK and MPV17 genes are associated with a severe clinical phenotype characterised by early-onset/neonatal ALF or rapidly progressive cholestasis and death before 12 months of age. A subset of MPV17 patients was amenable to LT. Consideration for LT in infantile ALF remains difficult and rapid genetic testing is advised.

Servicio profesional de monitorización ambulatoria de la presión arterial (MAPA) en farmacia comunitaria. Identificación de hipertensión no controlada de bata blanca / Ambulatory Blood Pressure Monitoring (ABPM) professional service in Community Pharmacy. White-coat UnControlled Hypertension identification

Varón de 68 años diagnosticado de hipertensión arterial hace 6 meses.Desde ese momento lo hemos incluido en nuestro Servicio Profesional Farmacéutico Asistencial deMedida y Control de la presión arterial (PA). Los sucesivos valores de PA obtenidos mediante medida aisladaen farmacia comunitaria (MAFC) han estado dentro de la normalidad.Actualmente el paciente tiene programada una intervención quirúrgica en el servicio de oftalmologíaque no se ha podido realizar por presentar sucesivos valores elevados de PA (≥180/100 mmHg) en consultamédica de atención primaria y especializada.El paciente refiere ponerse “muy nervioso” al acudir al médico. Sospechamos que los valores elevadosde PA en consulta son compatibles con un “efecto de bata blanca” (EBB).Acordamos con él y su médica de atención primaria la realización de una monitorización ambulatoriade presión arterial (MAPA) con el objetivo de descartar el EBB.Una vez obtenidos los resultados, identificar el fenotipo del paciente y observar que los valores de PAde la MAPA están dentro de la normalidad, se realiza una interconsulta con oftalmología.A la vista de los resultados el oftalmólogo decide realizar la operación, que se lleva a cabo con éxito. (AU)

68-year-old man diagnosed with hypertension 6 months ago. We have included it in our professionalpharmaceutical care service for measurement and control of blood pressure, obtaining values withinnormality.The patient has scheduled a surgical intervention in the ophthalmology service that could not beperformed due to successive high blood pressure values (≥180 / 100 mmHg) in a primary and specializedmedical consultation.He refers to getting “very nervous” when going to the doctor. We suspect that elevated office BP valuesare compatible with a “white coat effect” (WCE). We agreed with him and his primary care physician toperform ambulatory blood pressure monitoring (ABPM) in order to rule out EBB.Once the results have been obtained, identify the patient’s phenotype and observe that the ABPM BPvalues are within normality, an ophthalmology consultation is performed.In view of them, the ophthalmologist decides to perform the operation, which is carried out successfully. (AU)

Diagnostic Approach to Acute Liver Failure in Children: A Position Paper by the SIGENP Liver Disease Working Group.

Acute liver failure (ALF) is a clinical condition characterized by the abrupt onset of coagulopathy and biochemical evidence of hepatocellular injury, leading to rapid deterioration of liver cell function. In children, ALF has been characterized by raised transaminases, coagulopathy, and no known evidence of pre-existing chronic liver disease; unlike in adults, the presence of hepatic encephalopathy is not required to establish the diagnosis. Although rare, ALF has a high mortality rate without liver transplantation (LT). Etiology of ALF varies with age and geographical location, although it may remain indeterminate in a significant proportion of cases. However, identifying its etiology is crucial to undertake disease-specific management and evaluate indication to LT. In this position statement, the Liver Disease Working Group of the Italian Society of Gastroenterology, Hepatology and Nutrition (SIGENP) reviewed the most relevant studies on pediatric ALF to provide recommendations on etiology, clinical features and diagnostic work-up of neonates, infants and children presenting with ALF. Recommendations on medical management and transplant candidacy will be discussed in a following consensus conference.

Microbiological properties of poultry breast meat treated with high-intensity ultrasound.

Lactic acid, psychrophilic, and mesophilic bacteria, Escherichia coli, Salmonella spp. and Staphylococcus aureus were enumerated on chicken breasts after treatment with different high intensity ultrasound (frequency 40 kHz, intensity 9.6 W/cm-2) application times (0, 30, and 50 min) and packaging atmospheres (aerobic and vacuum) after a 7-day storage. The experiment was performed in commercial 7-week-old chicken breasts. Counts were performed prior to and immediately after ultrasonication, and on the 7th day of chill-storage. After sonication and storage, mesophiles, psychrophiles, LAB and S. aureus increased statistically. Psychrophiles decreased significantly under anaerobic packaging. There were no differences among ultrasonication times in terms of mesophiles, psychrophiles, LAB, E. coli and Salmonella spp. S. aureus numbers had a significant reduction after 50 min sonication. Under these experimental conditions, high-intensity ultrasound for 50 min is a control method of S. aureus and the anaerobic packaging reduces numbers of psychrophiles in chicken breast. The effect of ultrasound is only significant after the storage time.

Uroguanylin: a new actor in the energy balance movie.

Uroguanylin (UGN) is a potential target in the fight against obesity. The mature protein is released after enzymatic cleavage from its natural precursor, proUGN. UGN is mostly produced in the gut, and its production is regulated by nutritional status. However, UGN is also produced in other tissues such as the kidneys. In the past, UGN has been widely studied as a natriuretic peptide owing to its involvement in several different pathologies such as heart failure, cancer and gastrointestinal diseases. However, recent studies have suggested that UGN also acts as a regulator of body weight homeostasis because it modulates both food intake and energy expenditure. This ultimately results in a decrease in body weight. This action is mediated by the sympathetic nervous system. Future studies should be directed at the potential effects of UGN agonists in regulating body weight in human obesity.

Evaluación de la motilidad espermática usando dos crioprotectores / Evaluation of sperm motility using two cryoprotectants

La infertilidad, es un problema que afecta a una gran cantidad de parejas. Una de sus causas es la disminución de la calidad seminal debido, por ejemplo, a tratamientos gonadotóxicos. La criopreservación seminal es la técnica que permite conservar y almacenar espermatozoides sin que pierdan su capacidad fecundante; siendo esta una herramienta fundamental en reproducción asistida. El objetivo de este trabajo ha sido optimizar la técnica de criopreservación. Para ello se llevó a cabo un estudio, sobre muestras de pacientes en estudio por problemas de fertilidad, en el que se compararon dos medios de criopreservación (SpermCryo™All-round y CryoSperm™) y la aplicación o no de un baño en nitrógeno líquido a las muestras (previo a su almacenamiento); así como el efecto del tiempo que transcurre desde la eyaculación hasta el procesado sobre la calidad de la muestra. Las posibles variaciones fueron estudiadas con un analizador automático, mediante la realización de test pre- y post-congelación para comprobarla movilidad espermática

Infertility is a problem that affects a lot of couples. One of its causes is a decreased semen quality due to, for example, gonadotoxic treatments. The cryopreservation of human semen is the technique that allows sperm preserving and storing without losing their fertilizing capacity; being a fundamental tool in assisted reproduction. The aim of this study was to optimize the cryopreservation technique. To this end, a study carried out on samples of patients under study by fertility problems, in which two cryoprotectant media (SpermCryo™ All-round and CryoSperm™) and the execution or non-execution of an immersion of the samples in liquid nitrogen (before storage) were compared; and the effect of the time between ejaculation and the processing on the quality of the sample. Variations were studied with an automatic analyzer by performing pre- and post-thaw sperm motility tests. The results show no difference between the two cryoprotectants media, but seems to have a tendency to obtain better postthaw mobility with either depending on sample characteristics. Moreover, the liquid nitrogen bath had no apparent effects on post-thaw results. However, we must highlight the importance of time in the processing of semen samples once liquefied, to avoid decreased sperm quality. To improve post-thaw outcomes the key lies in the necessity to adjust the freezing protocol to the sample characteristics and a correct implementation of the protocol cryopreservation (selection and addition of cryoprotectant media...); favoring the management of infertility and the success of assisted reproduction techniques

Programa de preservación de la fertilidad en pacientes oncológicos / Fertility preservation program in cancer patientes

La infertilidad, es un problema que a día de hoy afecta a una gran cantidad de parejas. Una de las posibles causas es la disminución a nivel mundial de la calidad seminal debido a factores como pueden ser la alimentación, el consumo de alcohol y tabaco, altos niveles de estrés o factores ambientales. Además, el aumento de la incidencia de enfermedades tumorales en pacientes menores de 35 años, pone de manifiesto la necesidad de preservar la fertilidad de estos pacientes mediante técnicas de criopreservación. Es importante, que ésta se realice antes de que reciban ningún tipo de tratamiento (quimioterapia y radioterapia). En este trabajo, se llevará a cabo un estudio comparativo entre las muestras de varones que acuden al Hospital Materno Infantil por problemas de fertilidad y aquellos pacientes oncológicos que antes de iniciar el tratamiento realizan una criopreservación de la muestra seminal analizando las posibles variaciones de estas últimas muestras, mediante un test de post-congelación para comprobar la viabilidad de las muestras criopreservadas. Los parámetros que se van a analizar son: edad del paciente, volumen de la muestra, concentración espermática, porcentaje de espermatozoides móviles e inmóviles, móviles progresivos y espermatozoides tipo 'a'

The infertility, is a problem affecting a large number of couples today. One of the possible causes is decreasing around the world of the seminal ability due to factors such as food, alcohol and tobacco consumption, high levels of stress or environmental factors. In addition, the increase in the incidence of tumour diseases in patients younger than 35 years, highlights the need to preserve the fertility of these patients through cryopreservation techniques. It is important that this be done before they receive any kind of treatment (chemotherapy and radiotherapy). In this work, we will take place a comparative study between the samples of male couples who come to the mother and child Hospital for fertility problems and cancer patients before treatment carried out a Cryopreservation of spermatozoa by analysing possible variations of these latest samples, using a test of post-congelacion to check the viability of samples criopreserved. The parameters will be analyzed are: the patient's age, volume of sample, sperm concentration, percentage of progressive mobile and stationary, mobile sperm and sperm type 'a'

Collagen and matrix metalloproteinase-2 and -9 in the ewe cervix during the estrous cycle.

The cervical collagen remodeling during the estrous cycle of the ewe was examined. The collagen concentration determined by a hydroxyproline assay and the area occupied by collagen fibers (%C), determined by van Gieson staining, were assessed in the cranial and caudal cervix of Corriedale ewes on Days 1 (n = 6), 6 (n = 5), or 13 (n = 6) after estrous detection (defined as Day 0). In addition, the gelatinase activity by in situ and SDS-PAGE gelatin zymographies and matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9, respectively) expression by immunohistochemistry were determined. The collagen concentration and %C were lowest on Day 1 of the estrous cycle (P < 0.04), when MMP-2 activity was highest (P < 0.006) and the ratio of activated to latent MMP-2 trend to be highest (P = 0.0819). The MMP-2 activity was detected in 73% of the homogenized cervical samples, and its expression was mainly detected in active fibroblasts. By contrast, the MMP-9 activity was detected in 9% of the samples, and its scarce expression was associated with plasmocytes, macrophages, and lymphocytes. Matrix metalloproteinase-2 expression was maximal on Day 1 in the cranial cervix and on Day 13 in the caudal cervix and was lower in the cranial than in the caudal cervix (P < 0.0001). This time-dependent increase in MMP-2 expression that differed between the cranial and caudal cervix may reflect their different physiological roles. The decrease in the collagen content and increase in fibroblast MMP-2 activity in sheep cervix on Day 1 of the estrous cycle suggests that cervical dilation at estrus is due to the occurrence of collagen fiber degradation modulated by changes in periovulatory hormone levels.

Candidiasis oral en el paciente mayor / Oral candidosis in the older patient

La candidiasis o candidosis oral es la enfermedad infecciosa ocasionada por el crecimiento de las colonias de Cándida y la penetración de las mismas en los tejidos orales cuando las barreras físicas y las defensas del huésped se encuentran alteradas. Es una infección frecuente de la cavidad oral de los adultos de edad avanzada. Aunque la incidencia real se desconoce, se sabe que existe una prevalencia aumentada en ciertas ocasiones como ocurre en ancianos, en presencia de prótesis mucosoportadas, xerostomía o en patologías asociadas frecuentemente en los mayores. Los tipos clínicos más característicos son la forma seudomembranosa y la eritematosa (palatina y lingual). Pueden tener evolución aguda o crónica según la persistencia de los factores predisponentes. También son frecuentes procesos bucales comúnmente asociados: estomatitis protética, queilitis angular, glositis romboidal y lengua vellosa. La mayor parte de las candidiasis orales tienen un diagnóstico clínico, pero ha de confirmarse demostrando la penetración de la cándida en la mucosa oral, siendo el frotis la técnica de elección. Antes de comenzar el tratamiento, debemos estar seguros que se trata de una candidiasis oral, el tipo clínico y los factores predisponentes relacionados con la infección. Empezaremos siempre eliminando estos factores predisponentes, en el adulto mayor, la polifarmacología, la xerostomía, enfermedades crónicas y el uso de prótesis mucosoportadas son situaciones frecuentes que habrá que controlar. Instauraremos medidas higiénicas bucales y posteriormente si es necesario, utilizaremos fármacos antifúngicos, comenzando siempre con formas tópicas (AU)

Oral Candidiasis or Candidosis is the infectious disease caused by the growth of colonies of Candida and penetration in the oral tissues when physical barriers and host defenses are altered. It is the most common fungal infection of oral involvement. It is a common infection of the oral cavity in elderly adults. Although the true incidence is unknown, it is known that there is an increased prevalence in certain situations in the elderly: tissue-borne prosthesis, xerostomia or disorders frequently associated. The most characteristic clinical types are pseudomembranous and erythematous (palatal and lingual) form. They may have acute or chronic evolution as the persistence of predisposing factors. They are also frequent mouth commonly associated processes: denture stomatitis, angular cheilitis, rhomboid glossitis and hairy tongue. Most oral candidiasis have a clinical diagnosis, but must be confirmed by demonstrating penetration of candida on the oral mucosa, being the preferred technique smears. Before starting treatment, we must be sure that it is an oral candidiasis, clinical type and predisposing factors associated with infection. Always start eliminating these predisposing factors in the elderly, the polypharmacy, xerostomia, chronic diseases and the use of tissue-borne prostheses are common situations which must be controlled. We will initiate oral hygiene measures and then if necessary, use antifungaldrugs, always starting with topical forms (AU)

La endodoncia en los pacientes mayores / Endodontics in older patients

La endodoncia en el paciente geriátrico es un procedimiento cada vez más habitual en la clínica odontológica. Debemos, dadas las características médicas y dentales de muchos de estos pacientes mayores, planificar correctamente los procesos instrumentales y prever las posibles dificultades que puedan surgir motivadas por conductos estrechos, cámaras pulpares calcificadas etc. En la instrumentación de estos dientes, someteremos a los instrumentos a alto estrés por torsión, motivado por la amplia superficie de contacto entre el instrumento y las paredes, por lo que será totalmente necesario la realización de una preinstrumentación rotatoria y consideramos que es una buena opción el uso del movimiento recíproco. La preparación de los conductos debe dejar un remanente dentinario suficiente, mantenimiento del foramen apical y conicidad progresiva que facilita la administración de irrigantes a lo largo de la longitud de los conductos y las fuerzas de condensación para la obturación. Como irrigantes utilizamos, de forma general, el hipoclorito de sodio al 5% a lo largo de toda la fase instrumental; y una combinación del hipoclorito con un quelante durante la irrigación preobturación que activaremos con algún dispositivo ultrasónico o de aspiración apical negativa. La obturación la realizaremos con sistemas termoplásticos que nos aportan el mejor sellado tridimensional del sistema de conductos (AU)

Endodontics in geriatric patients is an increasingly common procedure in the dental clinic. Medical and dental characteristics of these patients makes us properly plan our procedures to anticipate possible difficulties as narrow ducts, calcifications etc. Our procedures should provide for the possible difficulties that may arise cause by narrow ducts, calcified pulp chambers, etc. During the instrumentation of these teeth, we submit a high stress of torsion to our instruments, motivated by the wide surface of contact between the instrument and the walls. For these reason, it will be absolutely necessary to make a rotary preinstrumentation, then, for the final instrumentation, we must consider the alternating rotation as a good option. The canal preparation should leave one sufficient dentin remnant, maintenance of the apical foramen and the proper taper to facilitate the administration of irrigants along the length of the ducts and the forces of condensation for obturation. Asirrigants we use, in general, sodium hypochlorite 5% throughout the entire instrumental phase; and a combination of hypochlorite with a chelat or for irrigation preobturation, in this phase we will activate with any ultrasonic device or with apical negative pressure. We will make the obturation with thermoplastic systems that provide us a completely three-dimensional sealing of the root canal system (AU)

Cervical changes in estrogen receptor alpha, oxytocin receptor, LH receptor, and cyclooxygenase-2 depending on the histologic compartment, longitudinal axis, and day of the ovine estrous cycle.

The aim was to investigate the histologic distribution of estrogen receptor α (ERα), oxytocin receptor (OxR), LH receptor (LHR), and cyclooxygenase-2 (COX-2) in the cervix of the ewe during the estrous cycle. Immunohistochemistry was performed in the cranial and caudal cervix of Corriedale ewes on Day 1 (n = 6), 6 (n = 5), or 13 (n = 6) after estrous detection (Day 0). The ERα proportional score (%ERα nuclei) was lower in the cranial cervix than in the caudal cervix, whereas the OxR and COX-2 immunostaining areas (%areas) were greater in the cranial cervix than in the caudal cervix (P < 0.04). The %ERα nuclei decreased from Days 1 to 13 in luminal epithelia, but increased from Days 1 to 6 or remained unchanged in stromata (P < 0.003). The %OxR area was higher on Day 6 than on Days 1 and 13 in the superficial glandular epithelium, and increased from Days 1 to 13 in the deep glandular epithelium (P < 0.04). The %LHR area increased during the estrous cycle in luminal epithelia and fold stroma (P < 0.004). The %COX-2 area was restricted to epithelia, and it was lower on Day 1 than on Days 6 and 13 in luminal epithelia (P < 0.05). Differences in ERα, OxR, LHR, and COX-2 between cranial and caudal cervical zones indicate different physiological functions, and their cyclic variations in the cervical epithelia, in contrast to little or no variations in the stroma, suggest a hormone-responsive driving role of epithelia in cervical function.

Expression of genes for oestrogen and progesterone receptors in the cervix of anoestrous ewes treated with gonadotrophin releasing hormone with or without progesterone priming.

The aim was to determine the oestrogens receptor alpha (ERα) mRNA and the binding capacity of oestrogens (ER) and progesterone receptor (PR) in the cervix of anoestrous ewes treated with gonadotrophin-releasing hormone (GnRH) with or without progesterone (P) priming, at the expected time of induced ovulation and early luteal phase. In Experiment 1, ewes were treated with P for 10 days (n=4), with nine micro-doses of GnRH followed by a GnRH bolus injection (n=4), or with P plus GnRH treatments (n=3), and tissues were harvested either without treatment (n=4), when P was removed, or 24h after the GnRH bolus injection. In Experiment 2, ewes were treated with the same GnRH or P plus GnRH treatments and tissues were harvested on Day 1 (n=12) or Day 5 (n=10) after the GnRH bolus injection. In the cranial cervix, the P treatment decreased and the GnRH treatment (after P treatment) increased the ERα mRNA, ER and PR concentrations (P<0.002). The ERα mRNA and ER concentrations were greater on Day 1, than on Day 5 in P plus GnRH treated ewes (P<0.0005). In the caudal cervix, lesser ERα mRNA, ER and PR concentrations than cranial cervix were found (P<0.0001). In conclusion, the ERα transcriptional activity and ER and PR binding capacity were strongly influenced by P and/or GnRH treatments in the cranial cervix, while the steroid receptors binding capacity remained unchanged in the caudal cervix of anoestrous ewes at the expected time of induced ovulation and early luteal phase.

A prospective 7-year survey on central venous catheter-related complications at a single pediatric hospital.

The aims of this study were to assess the incidence and risk factors of major central venous catheter (CVC)-related complications in a large cohort of children affected by oncological, hematological, or immunological diseases in a 7-year prospective observational study at a single center. Nine hundred fifteen CVCs were inserted in 748 children for a total period of 307,846 CVC-days. Overall, 298 complications were documented with a complication rate of 0.97/1,000 CVC-days: 105 mechanical complications (dislocations 0.30/1,000 CVC-days, ruptures 0.04/1,000 CVC-days), 174 infections (bloodstream infections 0.46/1,000 CVC-days, tunnel infections 0.10/1,000 CVC-days), and 19 thrombosis (0.06/1,000 CVC-days). Significant risk factors were: diagnosis of acute lymphoblastic leukemia (ALL) and age

Oestrogen and progesterone receptor binding capacity and oestrogen receptor alpha expression (ERalpha mRNA) along the cervix of cycling ewes.

The aim of the present work was to study the oestrogen receptor (ER) and progesterone receptor (PR) binding capacity and the oestrogen receptor alpha (ERalpha) mRNA concentration in cranial and caudal cervix during the ovine oestrous cycle. Cervical samples of synchronised Corriedale ewes were obtained on Day 1 (n=7), 6 (n=6) or 13 (n=7) after oestrus detection (Day 0). The ER and PR binding capacity by ligand-binding assay and the ERalpha mRNA concentration by solution hybridisation in both cranial and caudal zones of the cervix were determined. The ER and PR binding capacity were higher (P<0.005) on Day 1 than on Days 6 and 13 in both cranial and caudal zones. The ERalpha mRNA concentrations were higher (P<0.0001) on Day 1 than on Days 6 and 13 only in the caudal zone. The PR binding capacity and ERalpha mRNA concentration were higher (P<0.005) in the caudal than in the cranial zone on Day 1. The ER and PR expression in the ovine cervix varied during the oestrous cycle in agreement with the known upregulation exerted by oestrogen and downregulation exerted by progesterone. Differences in ER and PR expression along the longitudinal axis of the ovine cervix were found, reflecting histological and functional differences between the cranial and caudal zones.

Major childhood infectious diseases and other determinants associated with type 1 diabetes: a case-control study.

The objective of the study was to evaluate the association between infectious diseases and other events pertaining to childhood medical history and type 1 diabetes. A case-control study was carried out, taking as cases 159 type 1 diabetic patients (0-29 years) recorded from 1988 to 2000 within the population registry of the Pavia province (North Italy). As controls 318 non-diabetic subjects were matched by age and sex. A questionnaire was administered by standardised interviewers. Data were analysed by conditional logistic regression. Viral childhood diseases (OR 4.29; 95%CI 1.57-11.74) and bottle feeding (OR 1.83; 95%CI 1.08-3.09) were directly correlated to type 1 diabetes; an inverse correlation was found for vitamin D administration during lactation (0-14 years) (OR 0.31; 95%CI 0.11-0.86) and for history of scarlet fever in both sexes and age groups (OR 0.19; 95%CI 0.08-0.46). Most associations of the studied variables confirm already known findings. The significant inverse correlation of type 1 diabetes with scarlet fever history is a peculiar finding, the meaning of which is still obscure, although it has been recently described that streptococcal A infections are regulated by HLA class II alleles.

Corpus luteum life span and pituitary oestrogen and progesterone receptors in cyclic and gonadotrophin-releasing hormone-treated anoestrous ewes.

The present study investigated the pituitary oestrogen (ER) and progesterone (PR) receptor concentrations in ewes during the oestrous cycle in the breeding season (n = 19), and in anoestrous ewes treated with gonadotrophin-releasing hormone (GnRH) (n = 11) and anoestrous ewes treated with progesterone + GnRH (n = 11). The pituitary ER and PR concentrations at the expected time of ovulation and in the early and late luteal phases were measured by binding assay. The pattern of pituitary ER and PR concentrations in the progesterone + GnRH-treated ewes resembled the pattern found during the normal oestrous cycle, with ER and PR concentrations decreasing from the time of ovulation to the early luteal phase. In contrast, in ewes treated with GnRH alone, ER and PR concentrations increased in the early luteal phase, which may increase the inhibitory effects of steroid hormones on luteinising hormone secretion, ultimately leading to the development of subnormal luteal phases.

Experimentally induced subnormal or normal luteal phases in sheep: reproductive hormone profiles and uterine sex steroid receptor expression.

This study investigated if ewes expected to have subnormal luteal phases (SNLP) present a different pattern of uterine oestrogen receptor (ER) and progesterone receptor (PR) expression at the expected time of premature luteolysis. The concentrations of uterine ER, PR and ERalpha mRNA, and the steroid ovarian hormone were determined in anoestrous ewes treated with either gonadotrophin-releasing hormone (GnRH) to develop a SNLP (n = 16), or progesterone + GnRH to develop a normal luteal phase (NLP; n = 16). The ER, PR and ERalpha mRNA concentrations were measured using binding and solution hybridisation assays, while the hormone level concentration was measured by radioimmunoassay. In all ewes, a luteinising hormone- and follicle-stimulating hormone-synchronised surge was found. The SNLP group had lower preovulatory oestradiol levels than the NLP group. On Day 5, the SNLP group had lower progesterone levels, and higher uterine ER, PR and ERalpha mRNA concentrations than the NLP group. While in the SNLP group the receptor expression increased from Days 1 to 5, in the NLP group the receptor expression decreased. The results suggest that the induction of steroid receptor expression in the uterus and the hormonal environment found in the experimental SNLP group at the expected time of premature luteolysis may be involved in the mechanisms causing SNLP.

Differential estradiol effects on estrogen and progesterone receptors expression in the oviduct and cervix of immature ewes.

In order to study the effect of estradiol-17beta (E2) on estrogen (E) and progesterone (P) receptors expression in oviduct and cervix of lambs, their respective transcripts (ERalpha mRNA and PR mRNA) were determined by solution hybridization and the receptor proteins (ER and PR) by binding assays after E2 treatments. Lambs (n=4 in each group) were not treated or treated with one, two or three i.m. injections of E2 (1 microg/kg) at 24 h of interval. Tissues were obtained 12 or 24 h after the last E2 injection. Estradiol treatments increased ERalpha mRNA and PR mRNA concentrations in an organ-dependent manner: transitory in the oviduct while maintained in the cervix. The E2 effect on the oviductal and cervical ER and PR concentrations were biphasic, with an initial reduction of receptors content that was followed by restoration. The ER restoration in oviduct was earlier than in the cervix. In summary, this study shows that E2 treatments may exert an inductive effect in ERalpha mRNA and PR mRNA levels and a biphasic effect in ER and PR concentrations in oviduct and cervix of immature ewe. These E2 effects varied in timing and strength depending on the organ of the reproductive tract.

Chromosome analysis and mapping of ribosomal genes by one- and two-color fluorescent in situ hybridization in Hinnites distortus (Bivalvia: Pectinidae).

Metaphase chromosomes of the scallop Hinnites distortus were analyzed using Giemsa staining, chromosome measurements, silver staining, one- and two-color fluorescent in situ hybridization (FISH) ribosomal DNA (rDNA) probes, and 4',6-diamidino-2-phenylindole (DAPI) banding compatible with in situ hybridization. The karyotype (2n = 38) consists of three submetacentric-metacentric, one submetacentric, one subtelocentric-submetacentric, and 14 subtelocentric pairs. The 18S-28S rDNA maps at the centromeric level of two subtelocentric pairs, but not more than two nucleolus organizer region (NOR)-bearing chromosomes were transcriptionally active. The 5S rDNA seems to show a conventional tandem arrangement with a repeat unit of about 450 bp and it maps at the pericentromeric region of the long arm of one subtelocentric pair. Two-color FISH demonstrated that 18S-28S rDNA and 5S rDNA are not syntenic. Sequential FISH/Giemsa staining and subsequent chromosome pairing allow us to propose that pairs 9 and 12 carry the 18S-28S rDNA and pair 13 carries the 5S rDNA. All chromosomes are characterized as containing constitutive heterochromatin at the centromeric region. The data provided are the first contribution toward construction of the molecular karyotype of H. distortus and will be useful in assessing evolutionary relationships within scallops.