Pandemia SARS-CoV-2 y embarazo en el Hospital el Pino: un estudio descriptivo / SARS-CoV-2 pandemic and pregnancy in the El Pino Hospital: a descriptive study

INTRODUCCIÓN: En comparación con pandemias anteriores por otros virus, el SARS-CoV-2 ha afectado a la población obstétrica en menor número, por lo que registrar la historia clínica en pacientes con COVID 19 de nuestra población se hacía imperativo. METODOLOGÍA: Se realizó un estudio descriptivo transversal no analítico en la maternidad del Hospital El Pino. Se realizaron test PCR para SARS-CoV-2 entre abril y junio del 2020 a todas las usuarias con: (1) síntomas sugerentes de COVID 19, (2) antecedente de contacto estrecho o (3) que requiera hospitalización independientemente del motivo. En ellas se analizaron variables demográficas, evolución clínica y se estimaron "outcomes" obstétricos. RESULTADOS: Se tomaron 409 PCR y de éstas 365 en población obstétrica alcanzando en ella una incidencia acumulada de positividad de 19.7%. El 55.6% (40 pacientes) asintomáticas, 4.2% (9 pacientes) contacto estrecho y 40.3% (29 pacientes) sintomáticas. De estas pacientes la mayoría resulto ser de nacionalidad chilena (79.2%) con una prevalencia de HTA y DM tipo 1 o 2 entre el 8-10%. La media de la edad gestacional en la cual se realizó el diagnóstico fue en el 3er trimestre y la del momento del parto fue de 37.1 semanas (+/-3.9). Los síntomas más comunes presentados fueron tos, cefalea y mialgias; sin embargo, 7 requirieron ingreso a unidad de cuidados intensivos. En el seguimiento de usuarias asintomáticas 6,9% y 2,8% presentaron síntomas al día 7 y 14 respectivamente. CONCLUSIONES: La pandemia SARS-CoV-2 ha afectado la población obstetrica de igual manera, presentandose en su mayoría de forma asintomática, destacando la importancia del screening universal a la hora de hospitalizar nuestras usuarias y evitar transmisión intrahospitalaria.

INTRODUCTION: Compared to previous pandemics caused by other viruses, SARS-CoV-2 has affected the obstetric population in a smaller number, thus recording the medical history in patients with COVID 19 in our population became imperative. METHODOLOGY: A non-analytical, cross-sectional descriptive study was carried out in the maternity section of Hospital El Pino. PCR tests for SARS-CoV-2 were performed between April and June 2020 to all patients with: (1) symptoms suggestive of COVID 19, (2) a history of close contact or (3) requiring hospitalization regardless of motive. In them, demographic variables, clinical evolution were analysed and obstetric outcomes were estimated. RESULTS: 409 PCR were taken, raising a cumulative incidence of positivity of 19.7%. 55.6% (40 patients) were asymptomatic, 4.2% (9 patients) were close contact and 40.3% (29 patients) symptomatic. Within these patients, the majority turned out to be Chilean (79.2%) with a prevalence of HT and DM type 1 or 2 between 8-10%. The average of the gestational age at which the diagnosis was made was in the 3rd trimester and that of the moment of delivery was 37.1 weeks (+/- 3.9). The most common symptoms presented were cough, headache and myalgia, however, 7 patients required admission to the intensive care unit. In the monitoring stage of asyntomatic patients, 6.9% and 2.8% had symptoms on day 7 and 14 respectively. CONCLUSIONS: The SARS-CoV-2 pandemic has affected the obstetric population with no difference, with the majority presenting asymptomatic, fact which highlights the importance of universal screening when hospitalizing our users and avoiding intra-hospital transmission.

Diferenciación genética de las poblaciones de Felis catus en los municipios de Restrepo y Darién del Valle del Cauca, Colombia / Genetic differentiation of Felis catus populations in the municipalities Restrepo and Darién of Valle del Cauca, Colombia

Los genes implicados en los rasgos del pelaje de los gatos son útiles para el análisis de la estructura genética que presentan sus poblaciones. El objetivo de este trabajo fue determinar si existe diferenciación genética entre las poblaciones de gato doméstico de los municipios de Restrepo y Darién, los cuales se encuentran separados por el "Lago Calima". Para esto se estimaron las frecuencias alélicas de diversos marcadores del pelaje y se determinó si estas poblaciones presentaban diferencias significativas en su estructura génica o si se encontraban en equilibrio Hardy­Weinberg, además, se realizó una comparación con otras poblaciones inventariadas en el Valle del Cauca. Posteriormente, se llevó a cabo un análisis de componentes principales (ACP-Biplot), para conocer la correlación entre las frecuencias alélicas y los aportes de cada una de ellas a la variabilidad. También se realizó una prueba de Mantel para estimar si existía correlación entre los índices de fijación (FST) y las distancias geográficas. La prueba de Mantel mostró que el gen Orange es el único que muestra una correlación positiva entre los índices de diferenciación FST y las distancias geográficas. La comparación de las poblaciones de interés con las del Valle del Cauca, evidenció que los genes que aportaron más a la variabilidad fueron Dilution (19,08%), Long hair (16,09%), Agouti (16,06%) e Inhibitor (14,04%). Sin embargo, se encontró que las poblaciones de Restrepo y Darién tienen perfiles genéticos similiares y se comportan como una sola según los valores del equilibrio Hardy-Weinberg y los FST. Debido a que no hay diferencias significativas entre las poblaciones estudiadas, se concluye que, aunque el Lago Calima sea una barrera geográfica, no tiene un efecto significativo en la diferenciación genética entre las poblaciones de gatos de Restrepo y Darién.

The genes involved in cat coat traits are useful for the analysis of genetic structure within a population. The objective of this paper was to determine if any genetic differences exist among domestic cat populations from the municipalities of Restrepo and Darien, which are separated by Calima Lake. To analyze population structure, we estimated allelic frequencies of several coat markers, tested for alignment with the Hardy-Weinberg equilibrium, and compared our populations of interest to others from the Valle del Cauca region. Subsequently, we performed a principal components analysis (PCA-Biplot) to determine the correlation between allelic frequencies and their contributions to variability. A Mantel test was also used to estimate possible correlation among differentiation indexes (FST) and geographic distances. We performed the Mantel test on the Orange gene and identified a positive correlation among differentiation indexes (FST) and geographical distances. After comparing our populations of interest to others in the Valle del Cauca, we observed that the genes with the greatest contribution to variability were Dilution (19,08%), Long hair (16,09%), Agouti (16,06%) and Inhibitor (14,04%). However, we also found that the Restrepo and Darien populations of interest had similar genetic profiles, and aligned with the Hardy-Weinberg equilibrium, and the FST. Due to the absence of significant differentiation between the populations studied, we conclude that Calima Lake does not have a significant effect on any differentiation between the cat populations of Restrepo and Darien.

Genetic Differentiation of Colombian Populations of Anopheles darlingi Root (Diptera: Culicidae).

Anopheles darlingi Root is a primary vector of malaria in the neotropic region, a species not just highly anthropophilic but very efficient in transmitting Plasmodium species and considered the most important vector in the Amazon region. The main goal of this study was to determine the genetic structure of the A. darlingi populations using microsatellites (STR) in western and eastern regions of Colombia. DNA extraction was done with the cited protocol of band using the Genomic Prep™ cell and tissue isolation commercial kits. We used the STR reported by Conn et al (Mol Ecol Notes 1: 223-225, 2001). The analysis with STR proved there was a high genetic diversity and significant alterations of the Hardy-Weinberg equilibrium. The greatest genetic diversity was recorded in Mitu (Vaupes) (Na = 14, Ho = 0.520). The lowest was in Pueblo Nuevo (Cordoba) (Na = 12, Ho = 0.457). The eastern region and the Mitu (Vaupes) populations presented the highest number of primer alleles (Ap = 30; Ap = 13; Ap = 9), with variations between 0.010 and 0.097. The AMOVA revealed that the whole population underwent moderate genetic differentiation (F ST = 0.063, p < 0.05). The same differentiation was noticed (0.06 < F ST > 0.06, p < 0.05) with five of the six populations included in this job, and there was a low differentiation in the Las Margaritas (Santander) area (F ST = 0.02s3, p < 0.05). Our results suggest a slight positive correlation, which does not show a statistical significance between the geographic and genetic distances, probably suggesting that the moderate genetic differentiation found between pairs of populations does not need to be explained for the hypothesis of separation by distance.

Red Maca (Lepidium meyenii) did not affect cell viability despite increased androgen receptor and prostate-specific antigen gene expression in the human prostate cancer cell line LNCaP.

We examined whether aqueous extract of Lepidium meyenii (red Maca) could inhibit growth, potentiate apoptotic activity of two anticancer drugs Taxol and 2-methoxyestradiol (2ME) or change mRNA expression for the androgen target genes, androgen receptor (Ar) and prostate-specific antigen (Psa) in the human prostate cancer cell line LNCaP. Red Maca aqueous extract at 0, 10, 20, 40 or 80 µg/ml was added to LNCaP cells, and viability was evaluated by the MTS assay at 24 or 48 hr after treatment. Furthermore, LNCaP cells were treated with 80 µg/ml of red Maca plus Taxol or 2ME 5 µM and viability was assessed 48 hr later. Finally, LNCaP cells were treated with red Maca 0, 20, 40 or 80 µg/ml, and 12 hr later, mRNA level for Ar or Psa was assessed by real-time PCR. Treatment with red Maca did not affect viability of LNCaP cells. Apoptotic activity induced by Taxol and 2ME in LNCaP cells was not altered with red Maca treatment. Relative expression of the mRNA for Ar and Psa increased with red Maca 20 and 40 µg/ml, but not at 80 µg/ml. We conclude that red Maca aqueous extract does not have toxic effects, but stimulates androgen signalling in LNCaP cells.

Coat genetic markers of the domestic cat Felis catus (Felidae) from southwestern Colombia / Marcadores genéticos del pelaje del gato doméstico Felis catus (Felidae) del suroccidente colombiano

ABSTRACT Objectives. Establish the genetic profiles of cats from 12 neighboring municipalities in southwestern Colombia, in a town course from Pereira-Popayán. Estimate the degree of diversity, genetic structure, and quantify gene flow. Materials and methods. Were inventoried the phenotypic markers present in the pigmentation and structure of the coat of 1482 cats of the municipalities surveyed. Based on these phenotypic frequencies, allele frequencies, heterozygosity, Hardy-Weinberg equilibrium, F statistics and Nei genetic distances were calculated. A comparison was also made between genetic and geographic distance matrices to determine if there was a significant association between the two. Results. With the genetic profiles of the populations we estimated the degree of diversity. We found the populations in equilibrium for the S autosomal locus and for the O sex-linked locus. We found a low level genetic structure, and it was determined that there was no significant correlation between the genetic and geographic distance matrices among populations. Conclusions: These findings can be explained on the basis of the processes of human displacement for this region, due to the fact that the establishment of feline populations in these municipalities originated during the same historical period. Identical genetic profiles are shared as a result of colonization events, and due to possible continued migration among these populations.

RESUMEN Objetivos. Determinar los perfiles genéticos de los gatos de 12 municipios contiguos del suroccidente colombiano en un trayecto Pereira-Popayán; estimar el grado de diversidad, estructura genética y cuantificar el flujo de genes. Materiales y métodos. Se inventariaron los marcadores fenotípicos presentes en la pigmentación y estructura del pelaje de 1482 gatos de los municipios estudiados, con base en estas frecuencias fenotípicas se calcularon frecuencias alélicas, heterocigosidad, equilibrio Hardy-Weinberg, el estadístico FST y distancias genéticas de Nei. También se realizó una comparación entre matrices de distancia genética y geográfica para determinar si existía asociación significativa entre las dos. Resultados. Con los perfiles genéticos de las poblaciones se estimó el grado de diversidad, se halló en equilibrio Hardy-Weinberg a las poblaciones para el locus autosómico S y el locus ligado al sexo O. Se encontró bajo nivel de estructura genética y se determinó que no existe correlación significativa entre las matrices de distancia genética y geográfica entre poblaciones. Conclusiones. Estos hallazgos se pueden explicar con base en los procesos de desplazamiento humano para esta región, debido a que el establecimiento de poblaciones felinas en estos municipios se originó en el mismo período histórico. Se comparten perfiles genéticos idénticos como resultado, posiblemente, de eventos de colonización y migración continua entre estas poblaciones.

Prevalencia de genotipos del papiloma virus en mujeres de Cuenca / Prevalence of genotypes of papilloma virus in women in Cuenca

Objetivos: Determinar la prevalencia de los genotipos del papiloma virus de alto y bajo grado oncogénico; relacionar con los grupos de edad, con los genotipos que evitan las dos vacunas existentes y con los fatores de riesgo para cáncer cervical uterino. Metodología: Estudio epidemiológico, observacional, transversal. La muestra aleatorizada fue de 500 mujeres de la zona urbana de la ciudad de Cuenca. Se efecturaon dos tomas del cuello uterino. Se utilizó el método de reacción de cadena de polimerasa y el estudio citopatológico Papanicolaou. Resultados: La prevalencia de los genotipos del papiloma virus de alto grado oncogénico fue del 35.9% y para bajo grado el 14.3%. La prevalencia de las alteraciones citológicas, Papanicolaou, fue del 16%. Estas prevalencias fueron más frecuentes en los grupos de 30 a 39 años y 40 a 50 años. Por inferencia, la cobertura de la vacuna Cervarix sería del 8% y para el Gardasil del 13%. Fueron factores de riesgo los relacionados con el número y frecuencia de relaciones sexuales, número de compañeros sexuales. Conclusiones. La prevalencia de los genotipos fue alta y la cobertura de las vacunas baja.

OBJECTIVES. To determine the prevalence of papilloma virus genotypes of high and low oncogenic grade, to relate to age groups, with genotypes that avoid both existing vaccines and the risk factors for uterine cervical cancer. METHODS. Epidemiological, observational, cross-sectional study. The random sample was 500 women from the urban area of Cuenca city. Two samples of the cervix were taken. Polimeraza chain reaction method and cytopathology Papanicolaou study were used. RESULTS. The prevalence of papilloma virus genotypes of high oncogenic grade was 35,9% and 14,3% low oncogenic grade. The prevalence of cytologic abnormalities, Papanicolaou, was 16%. This prevalence was more frequent in the group of 30-39 years, and 40-50 years. By inference, the coverage of the Cervarix vaccine would be 8% and 13% for Gardasil. Risk factors were related to the number and frequency of intercourse and number of sexual partners. CONCLUSION. The prevalence of genotypes was high and the vaccine coverage was low.

Development of a low-color, color stable, dual cure dental resin.

UNLABELLED: Dual-cure (DC) resins are mainly used as cements due to high initial color (generally yellow) and large color shift (ΔE*) after polymerization as compared to light-cured resins. However, even as cements, this color shift is clinically unacceptable, especially when used to cement thin veneers. OBJECTIVE: To develop a novel DC initiator system with both lower initial color (less yellow, i.e., whiter) and smaller ΔE*. METHODS: The effect of using an allyl thiourea (T)/cumene hydroperoxide (CH) self-cure (SC) initiator system in combination with a photo-co-initiator, p-octyloxy-phenyl-phenyl iodonium hexafluoroantimonate (OPPI), in a commercial DC resin cement (PermaFlo DC, Ultradent Products, Inc.) was investigated. Initial color and ΔE* were assessed for 6 weeks in vitro under accelerated aging conditions (75°C water bath). Rockwell15T hardness was used to assess degree of cure (DoC) and the three-point bending test was used to assess mechanical properties. RESULTS: PermaFlo DC (control) was significantly harder than all experimental groups without OPPI but had up to three times higher initial color and four times greater color shift (ΔE*=27 vs. 8). With OPPI, hardness in the experimental groups increased significantly and several were comparable to the controls. Initial color and ΔE* increased slightly (ΔE*=9), but was still 3 times less than that of PermaFlo DC. DC samples containing OPPI had comparable modulus and ultimate transverse strengths to those of the controls. CONCLUSIONS: DC resins that use the T/CH initiator system are weaker but have extremely low color and ΔE*. The addition of OPPI increases DoC and mechanical properties to clinically acceptable levels and maintains extremely low color and ΔE*. SIGNIFICANCE: With this novel initiator system, DC resins potentially can now have comparable color and color stability to light-cure resins and be used in broader esthetic dental applications to improve color stability and reduce shrinkage stress in restorative composites.

Simplificando el tratamiento quirúrgico de la sonrisa gingival / Simplifying the surgical treatment of the gummy smile

La sonrisa es una de las expresiones más comunes del ser humano como muestra de satisfacción, alegría o felicidad. Algunas personas al sonreír muestran la encía por encima de la base de los dientes, lo que ocasiona un aspecto antiestético. Esta situación repercute negativamente en el individuo, ya que altera la espontaneidad de la expresión facial. En el origen de la sonrisa gingival intervienen factores dentales, esqueléticos y de tejidos blandos. Proponemos una variante quirúrgica innovadora para corregir la sonrisa gingival en pacientes con alteraciones de los tejidos blandos. Se intervinieron quirúrgicamente 15 mujeres de entre 17 y 38 años de edad. En 7 de ellas se practicó rinoplastia además de corrección de la sonrisa gingival. Las 8 restantes se sometieron exclusivamente a corrección de la sonrisa gingival. Las 15 pacientes intervenidas mejoraron su apariencia y autoestima. No hubo complicaciones o problemas relacionados con la herida quirúrgica. Los resultados han sido permanentes a medio y largo plazo. La sonrisa delas pacientes es normal, mejoró su expresión y se conservaron la sensibilidad y la función muscular La técnica propuesta es sencilla y de fácil reproducción; el tiempo quirúrgico estimado es de una hora y la operación es de tipo ambulatoria. Suele ser bien tolerada por los pacientes y no altera sus actividades cotidianas (AU)

Human beings possess a highly expressive face, there are people who when smile, show the gums above the base of the teeth causing an antiesthetic aspect. This situation has negative repercussions altering the spontaneity of the facial expression. In the etiology of the gingival smile, dental, skeletal and soft tissue factors take part. We present a new surgical variant that offers an excellent alternative for the treatment of the gingival smile with soft tissue alterations. Fifteen female patients were treated; 7 had rhinoplasty together with correction of gingival smile; the remaining 8 patients were exclusively gingival smile surgery; agesvaried from 17 to 38 years. All the treated improved their appearance and selfesteem. There were no complications or problems related to the wound. The results are permanent in the medium and long term and the patient’s smile is normal because of improvement of their expression, maintaining muscular function and sensitivity. The proposed technique is simple, easily reproducible, ambulatory and with an average operating time of one hour. It is well tolerated by the patients since it does not interfere with their daily activities (AU)

Rubber-toughening of dimethacrylate dental composite resin.

UNLABELLED: Dimethacrylate dental composite resins exhibit inherently low toughness. Toughening of these materials may reduce the incidence of marginal and bulk fracture of composite restorations. OBJECTIVE: To determine if dimethacrylate dental restorative materials can be rubber-toughened, and if so, to identify a possible mechanism. METHODS: A filler composed of aggregates of polybutadiene/silica as well as irregularly-shaped silica slabs was produced by mixing silica with polybutadiene in dichloromethane. The dried filler was subsequently ground and sieved to < 25 microm. Polybutadiene/silica ratios were varied from 0:1 (control) to 0.5:1. EDAX analysis verified the composition of the complex filler. Filler was added to a bis-GMA/bis-EMA/TEGDMA resin system and fractured in three-point bend test mode at a crosshead speed of 1 mm/min. In addition, 1 bar was fractured at a crosshead speed of 0.001 mm/min to identify a possible mechanism for toughening. RESULTS: In specimens fractured at 1 mm/min, flexural modulus is increased or maintained and flexural strength and energy to break increase as the amount of polybutadiene in the aggregates increases. Cavitation of high-rubber-containing aggregates is demonstrated. In the one specimen fractured at 0.001 mm/min, a marked increase in size of high-rubber-containing aggregates along with severe shear damage in the surrounding matrix is shown, suggesting that cavitation with subsequent absorption of energy during shear yielding is the likely mechanism behind the increase in energy to break in bars fractured at 1 mm/min. SIGNIFICANCE: These results indicate that dimethacrylate dental composite materials can be rubber toughened, which may potentially reduce marginal and bulk fractures of composite restorations, and consequently extend their service lifetime.

Permanent Genetic Resources added to Molecular Ecology Resources database 1 January 2009-30 April 2009.

This article documents the addition of 283 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Agalinis acuta; Ambrosia artemisiifolia; Berula erecta; Casuarius casuarius; Cercospora zeae-maydis; Chorthippus parallelus; Conyza canadensis; Cotesia sesamiae; Epinephelus acanthistius; Ficedula hypoleuca; Grindelia hirsutula; Guadua angustifolia; Leucadendron rubrum; Maritrema novaezealandensis; Meretrix meretrix; Nilaparvata lugens; Oxyeleotris marmoratus; Phoxinus neogaeus; Pristomyrmex punctatus; Pseudobagrus brevicorpus; Seiridium cardinale; Stenopsyche marmorata; Tetranychus evansi and Xerus inauris. These loci were cross-tested on the following species: Agalinis decemloba; Agalinis tenella; Agalinis obtusifolia; Agalinis setacea; Agalinis skinneriana; Cercospora zeina; Cercospora kikuchii; Cercospora sorghi; Mycosphaerella graminicola; Setosphaeria turcica; Magnaporthe oryzae; Cotesia flavipes; Cotesia marginiventris; Grindelia Xpaludosa; Grindelia chiloensis; Grindelia fastigiata; Grindelia lanceolata; Grindelia squarrosa; Leucadendron coniferum; Leucadendron salicifolium; Leucadendron tinctum; Leucadendron meridianum; Laodelphax striatellus; Sogatella furcifera; Phoxinus eos; Phoxinus rigidus; Phoxinus brevispinosus; Phoxinus bicolor; Tetranychus urticae; Tetranychus turkestani; Tetranychus ludeni; Tetranychus neocaledonicus; Tetranychus amicus; Amphitetranychus viennensis; Eotetranychus rubiphilus; Eotetranychus tiliarium; Oligonychus perseae; Panonychus citri; Bryobia rubrioculus; Schizonobia bundi; Petrobia harti; Xerus princeps; Spermophilus tridecemlineatus and Sciurus carolinensis.

Dihydrotestosterone influenced numbers of healthy follicles and follicular amounts of LH receptor mRNA during the follicular phase of the estrous cycle in gilts.

The present experiments were conducted to determine androgenic effects on numbers, health, and amounts of gonadotropin receptor mRNA in late developing follicles of gilts. Gilts (n=5 per group) received daily injections of one of the following treatments on days 13-16 or days 13-18 of the estrous cycle: corn oil, 5alpha-dihydrotestosterone (DHT, 10 mg), flutamide (1.5 g, an androgen receptor inhibitor), DHT (10 mg) plus flutamide (1.5 g), testosterone (10 mg), and testosterone (10 mg) plus flutamide (1.5 g). Ovarian follicles > or =5 mm in diameter were evaluated on day 17 or 19, 24 h after receiving the last treatment dose. Follicles were classified as healthy (H), moderately atretic (MA), or very atretic (VA). Treatment with DHT increased (P<0.05) the numbers of H follicles relative to control gilts on days 17 and 19. DHT administration from days 13 to 16 diminished (P<0.05) the amounts of LH receptor (LHR) mRNA in H follicles from day 17 (relative amounts: 1.45+/-0.33 and 2.72+/-0.33 for DHT- and vehicle-treated gilts respectively). The effects of DHT on numbers of H follicles and LHR mRNA were not observed in gilts receiving DHT plus flutamide. Androgens did not influence numbers of MA, VA, and total follicles, or follicular estradiol-17beta concentrations and amounts of FSHR mRNA. Treating gilts with DHT during follicular recruitment and selection did not induce changes in the numbers of total follicles > or =5 mm, but rather increased the numbers of healthy follicles in this follicular population in association with decreased amounts of LHR mRNA.

Steroidogenic changes and steady state amount of messenger RNA encoding steroidogenic enzymes, gonadotropin receptors and cell-death signalling in the dominant ovarian follicle during estradiol-induced atresia in cattle.

Changes in steroidogenic function and associated gene expression were characterized in dominant ovarian follicles (DF) of cattle where follicles were induced to become atretic by systemic administration of estradiol benzoate (EB). In experiment 1, follicular fluid (FF) steroid concentrations in the DF were measured at 12-hourly time points for 48 h in heifers treated with 1 mg EB i.m./500 kg body weight (EB; n=20) as compared with untreated controls (C; n=19). Treatment with EB promoted a transient reduction in circulating FSH, a rapid (12 h) and sustained reduction in FF estradiol, a rapid (12 h) but transient reduction in FF progesterone and a delayed (36 h) increase in FF testosterone concentrations. In experiment 2, whole follicular wall tissue was collected from DF of mature non-lactating cows allocated to a 0 h control group (0 HC: n=7), a 24h control group (24 HC; n=7) or an EB-treated group where tissue was collected 24 h after administration of 1 mg EB i.m./500 kg body weight (EB; n=8). As for experiment 1, EB promoted a transient reduction in circulating FSH, a pronounced reduction in FF estradiol and a smaller but significant reduction in FF progesterone concentrations. Semi-quantitative RT-PCR on follicular wall tissue revealed that the loss in estrogen activity at 24 h after EB was associated with two-fold reduction in aromatase mRNA, with an apparent acceleration in loss of 17alpha-hydroxylase mRNA. Expression of genes for gonadotropin receptors (LHR and FSHR) and a cell-death signalling pathway (Fas antigen and Fas ligand) were unchanged during the initial 24h of EB-induced atresia. These results suggest that EB initiates atresia in dominant ovarian follicles through a rapid suppression of follicular estradiol synthesis, an effect associated with down-regulation of the aromatase gene. A transient suppression in circulating FSH following administration of EB appears to have initiated these events, and it is suggested that subsequent processes involved in atresia follow this loss in estrogenic function.

Deficient expression of monoamine oxidase A in the endometrium is associated with implantation failure in women participating as recipients in oocyte donation.

Successful implantation depends both on the quality of the embryo and on the endometrial receptivity. The latter depends on progesterone-induced changes in gene expression, a process that has been characterized by microarray analysis. One of the genes whose transcription appears to be enhanced during the receptive period is monoamine oxidase A (MAO-A). Our first objective was to confirm the increased expression of MAO-A in the endometrium during the receptive phase of spontaneous normal cycles using real time PCR and immunofluorescence. The second objective was to examine the endometrial expression of MAO-A during the receptive phase induced by exogenous estradiol (E(2)) and progesterone in patients whose endometrium was shown to have been either receptive or non-receptive to embryo implantation in repeated cycles of oocyte donation. Results showed that MAO-A transcript levels increased between the pre-receptive (LH+3) and receptive phase (LH+7) in all spontaneous cycles examined, with a median increase of 25-fold. Immunofluorescent labelling demonstrated MAO-A localization to the glandular and luminal epithelium with an increasing positive score between LH+3 and LH+7. Conversely, prior failure of embryo implantation was associated with a 29-fold decrease in MAO-A mRNA levels and a substantial reduction in MAO-A protein immunofluorescent label score. These results show a strong association between endometrial receptivity and MAO-A expression in the endometrial epithelium, suggesting an important role for this enzyme in normal implantation.

Androgens in female pig reproduction: actions mediated by the androgen receptor.

Androgens have potential actions in almost all the organs of males and females. In females, most organs contain some tissues with cells that have androgen receptors. Androgens can regulate cellular functions by binding to androgen receptors or be converted to other hormones. For example, testosterone can bind to the androgen receptor or be aromatised to oestradiol. Treating animals with testosterone, therefore, might elicit some androgenic and oestrogenic effects. Alternatively, testosterone can be converted to other androgens, which in turn, have more or less affinity with the androgen receptor and these new metabolites may or may not be aromatised to oestrogens. This review will highlight the roles of androgens in female mammals other than those as a substrate for oestrogen, with particular emphasis on the actions of the androgen receptors in uteri and ovaries of pigs. Utilising small dosages of an androgen receptor agonist, DHT (5alpha-dihydrotestosterone) we have observed that some uterine functions were inhibited while ovarian follicular development was augmented. These inhibitory and stimulatory effects of androgen therapy on reproductive organs can potentially be balanced to enhance ovulation rate and litter size in gilts and sows. Perhaps after future experimentation, new uses of androgens or anti-androgens could improve additional aspects of sow performance not presently under consideration.

Effects of cadmium on Na+ transport in the isolated skin of the toad Pleurodema thaul.

Cadmium ions applied to either (outer or inner) surface of the isolated toad skin dose-dependently increased the short-circuit current (SCC), the potential difference (V) and the active sodium conductance (G(Na)) in the concentration range 0.07-0.50mM. Maximal stimulatory effect was over 30% with an EC(50) of about 0.1mM. The effect of the highest concentration used (0.75mM) decreased considerably, and when it was applied to the inner surface (10 experiments), induced between 30% and 40% inhibition of the electric parameters in four experiments. Pretreatment with amiloride inverted the stimulatory effect of externally applied Cd(2+), suggesting competitive action on the apical Na(+) channel. The effect of noradrenaline (NA) was increased after outer application of Cd(2+) and decreased after inner application of the metal: the latter effect might be due to cadmium inhibition of the activity of Na(+),K(+)-ATPase. On the other hand, pretreatment with amiloride was followed by partial although transient reversal of its effects by serosal Cd(2+), which might be explained by action of cadmium on cytoplasmic lysine residues concerned with Na(+) channel gating. The amiloride test showed that the increment of the electric parameters was due principally to stimulation of the driving potential for Na(+) (V-E(Na(+))) and that inhibition was accompanied by a reduction in the V-E(Na(+)) and by a significant decrease in skin resistance indicating possible disruption of membrane or cell integrity. These data are in favor of the possibility that externally applied Cd(2+) activates toad skin ion transport, partly by increasing apical sodium conductance and also by stimulating the V-E(Na(+)), and that internally applied Cd(2+), with easier access to membrane and cellular constituents, may inhibit the sodium pump.

Iron affects the structure of cell membrane molecular models.

The effects of Fe(3+) and Fe(2+) on molecular models of biomembranes were investigated. These consisted of bilayers of dimyristoylphosphatidylcholine (DMPC) and of dimyristoylphosphatidylethanolamine (DMPE), classes of phospholipids located in the outer and inner moieties of cell membranes, respectively. X-ray studies showed that very low concentrations of Fe(3+) affected DMPC organization and 10(-3)M induced a total loss of its multilamellar periodic stacking. Experiments carried out with Fe(2+) on DMPC showed weaker effects than those induced by Fe(3+) ions. Similar experiments were performed on DMPE bilayers. Fe(3+) from 10(-7)M up to 10(-4)M had practically no effect on DMPE structure. However, 10(-3)M Fe(3+) induced a deep perturbation of the multilamellar structure of DMPE. However, 10(-3)M Fe(2+) had no effect on DMPE organization practically. Differential scanning calorimetry measurements also revealed different effects of Fe(3+) and Fe(2+) on the phase transition and other thermal properties of the examined lipids. In conclusion, the results obtained indicate that iron ions interact with phospholipid bilayers perturbing their structures. These findings are consistent with the observation that iron ions change cell membrane fluidity and, therefore, affect its functions.

Distribution and changes in amounts of the androgen receptor in the pig uterus during the estrous cycle, early pregnancy and after treatment with sex steroids.

Two experiments were performed to examine the expression of the androgen receptor (AR) gene in the pig uterus. In experiment 1, immunohistochemistry (IHC) was used to determine the distribution of the AR in uterine tIssue of pigs when collected at the first day of estrus (day 0) and the mid-luteal phase (day 12) of the estrous cycle, or early pregnancy (day 12, n=4 gilts per group). In experiment 2, AR immunostaining and AR mRNA in uterine tIssue were compared among ovariectomized gilts (n=4 per group) following treatment for 4 days with daily injections of: (1) progesterone (2 mg/kg bodyweight (BW)), (2) estradiol-17beta (E(2,) 2 micro g/kg BW), (3) E(2) plus progesterone (same dosages as 1 and 2 combined), (4) 5alpha-dihydrotestosterone (DHT, 7 micro g/kg BW), or (5) vehicle (corn oil). Data were analyzed using ANOVA. In experiment 1, nuclear staining for AR in luminal and glandular epithelia was strong and did not differ in intensity between the two locations. Immunostaining of AR in the myometrium was less (P<0.001) intense than in the luminal and glandular epithelia. Nuclei of stromal cells contained AR immunostaining that varied in intensity from strong (mainly in subepithelial stroma) to weak or no staining. Stages of the estrous cycle or early pregnancy did not influence AR immunostaining in the endometrial epithelia and myometrium. In experiment 2, immunostaining of AR in glandular and luminal epithelia and myometrium of ovariectomized gilts treated with vehicle or DHT was less (P<0.05) than in gilts treated with E(2), progesterone, or E(2) plus progesterone. Immunostaining of AR did not differ between ovariectomized gilts treated with vehicle or DHT, or between gilts treated with E(2), progesterone, or E(2) plus progesterone. In both experiments, intensity of AR immunostaining was greater in glandular epithelium located at the adluminal region compared with glandular epithelium located at the basal region of the endometrium. Competitive reverse-transcription PCR (RT-PCR) indicated a stimulatory effect (P<0.01) of E(2) on amounts of AR mRNA in whole endometrium. This increase in AR mRNA after E(2) treatment was not detected when E(2) was combined with progesterone. Endometrial AR mRNA was not influenced by DHT or progesterone relative to vehicle-treated gilts. In conclusion, immunoreactive AR is mainly present in luminal and glandular epithelia of the pig uterus and to a lesser extent in the myometrium, and does not change significantly during the estrous cycle or early pregnancy. Expression of the AR gene in the pig endometrium and myometrium appears to be regulated by E(2) and progesterone.

Androgen receptor and follicle-stimulating hormone receptor in the pig ovary during the follicular phase of the estrous cycle.

Follicle-stimulating hormone (FSH) is an important regulator of follicular development. Some effects of FSH on ovarian follicles might be enhanced by androgens. The main objectives of the present study were to examine expression of the androgen receptor (AR) and FSH receptor (FSHR) in late developing follicles in pigs. Ovaries were collected from gilts on days 13, 15, 17, and 19 of the estrous cycle (day 0 = first day of estrus, n = 4 gilts/day), a period coincident with the follicular phase. One ovary was processed for immunohistochemistry (IHC) of AR. Samples of surface wall from the largest follicles (4-5 per gilt) were dissected from the other ovary, pooled and processed for determination of AR and FSHR mRNAs using reverse transcription-polymerase chain reaction (RT-PCR). Intense AR immunostaining was present in nuclei of granulosa cells of preantral and antral follicles. AR immunoreactivity was also present in the nuclei of oocytes. Weak staining for AR was observed in cells of the theca interna, ovarian surface epithelium, and in most cells of the ovarian stroma. Relative amounts of immunoreactive AR in granulosa cells of late developing follicles, or small antral follicles (< 2 mm), did not differ between days 13, 15, 17, and 19. However, amounts of AR in granulosa cells of small antral follicles was greater (P < 0.05) than in the largest follicles present in the same ovary. The relative amounts of AR mRNA in tissue from the largest follicles on days 13, 15, 17, and 19 did not differ; however, amounts of FSHR mRNA in the same follicles were not different between days 13, 15, and 17, but decreased (P < 0.05) by day 19. Results indicate that during the follicular phase in gilts, the AR protein is mainly present in granulosa cells. Relative amounts of AR protein in granulosa cells and mRNA in walls of late developing follicles did not significantly change from day 13 to 19; however, amounts of FSHR mRNA decreased in preovulatory follicles by day 19 of the estrous cycle.

Increased ovulation rate in gilts treated with dihydrotestosterone.

Treatment with testosterone increases ovulation rate in pigs. The present study was conducted to examine the effects of 5alpha-dihydrotestosterone (DHT), a non-aromatizable androgen receptor ligand, on ovulation rate and amounts of androgen receptor and FSH receptor mRNAs in postpubertal gilts. In Expt 1, ovulation rate in response to daily i.m. injections of 0, 6, 60 or 600 microg DHT kg(-1) body weight from day 13 of the oestrous cycle (day 0 = day 1 of oestrus) to the following oestrus increased with each dose of DHT (P < 0.05). The mean increase in number of corpora lutea ranged from approximately three to 17 over the three dosages of DHT. In Expt 2, gilts treated daily with 60 microg DHT kg(-1) body weight during the early follicular phase (from day 13 to day 16), coincident with follicular recruitment, or the late follicular phase (day 17 to oestrus), had higher (P < 0.05) rates of ovulation compared with gilts that received vehicle, and were not different from gilts treated with DHT from day 13 to oestrus. Percentage recovery of day 3 embryos was not altered when gilts were treated from day 13 to day 16 or from day 17 to oestrus; however, treatment of gilts with DHT from day 13 to oestrus decreased recovery of day 3 (Expt 1) or day 11 (Expt 2) conceptuses. Daily administration of 6 microg DHT kg(-1) body weight to gilts from day 13 of the oestrous cycle to the following oestrus (Expt 3) did not affect the relative amounts of androgen receptor mRNA, but increased (P < 0.05) the amounts of FSH receptor mRNA in preovulatory follicles as determined by RT-PCR. The results of these experiments indicate that androgens may regulate ovulation rate in gilts. One of the roles of androgens might be regulation of the amounts of FSH receptor mRNA in ovarian follicles.

Molecular identification of evolutionarily significant units in the Amazon River dolphin Inia sp. (Cetacea: Iniidae).

The Amazon river dolphin, genus Inia, is endemic to the major river basins of northern South America. No previous studies have focused on the genetic structure of this genus. In this work, 96 DNA samples from specimens of this genus were collected in the Orinoco basin (four rivers), the Putumayo River, a tributary of the Colombian Amazon and the Mamoré, and the Tijamuchí and Ipurupuru rivers in theBolivian Amazon. These samples were used to amplify a fragment of 400 bp of the mitochondrial DNA (mtDNA) control region. In addition, 38 of these samples were also used to sequence 600 bp of the mitochondrial cytochrome b gene. The analysis of the population structure subdivision with an analysis of molecular variance (AMOVA) revealed important aspects about the genetic structure of Inia groups fromthese three geographically separate regions. By comparing the control region DNA and cytochrome b sequences, distinct types of nonshared haplotypes were observed. The net genetic divergence of control region sequences was 6.53% between the Orinoco and Bolivian rivers, 5.32% between the Putumayo and Bolivian rivers, and 2.50% between the Orinoco and Putumayo rivers. For the cytochrome b gene, these values were 2.48%, 2.98%, and 0.06%, respectively. The nucleotide sequences were analyzed phylogenetically using several genetic distance matrices and applying neighbor-joining, maximum likelihood, and maximum parsimony procedures. The results support the proposal to subdivide the Inia genus into at least two evolutionarily significant units: one confined to the Bolivian river basin and the other widely distributed across the Amazon and Orinoco basins.