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Electrophysiological aspects of human sphincter function.

In order to investigate the electrophysiology of the human internal anal sphincter and two current concepts of sphincter function, simultaneous manometric and electrical recordings were made from circular smooth muscle of the internal anal sphincter in the resting state and during reflexly induced sphincter relaxation. Three groups were studied: seven normal subjects, 25 patients with functional bowel disease, and seven patients with external sphincter paralysis due to spinal cord lesions. In the resting state slow waves of alternating potential (basic electrical rhythm or BER) were recorded in all subjects. Two types of waves were present, a constant sinusoidal pattern or a spindleshaped pattern. Either pattern was consistent for a given individual. Frequency of BER in the internal sphincter was higher than that recorded in any other gastrointestinal muscle. Our findings indicate that the BER recorded from the internal anal sphincter originates in this muscle. This activity may represent a specialized feature of sphincteric muscle since BER cannot be recorded from isolated nonsphincteric circular muscle. Reproduction of the two patterns of BER by an electronic model suggests that BER, as recorded by this technique, results from a summation of a number of electrically active cells in contact with the recording electrodes. Inhibition of BER occurred when sphincter relaxation was reflexly induced by rectal distension. Both inhibition of BER and degree of sphincter relaxation were proportional to the strength of rectal stimulation, suggesting that strength of stimulus determines the number of active cells which are inhibited. The associations of high frequency of BER with high resting pressure, and of inhibition of BER with sphincter relaxation suggests that maintenance of sphincter tone is an active process that is governed by BER.

Adolescent↗

Experimental reproduction of pneumonia in gnotobiotic pigs with porcine respiratory coronavirus isolate AR310.

The pathogenicity of porcine respiratory coronavirus (PRCV) isolate AR310 was determined for gnotobiotic pigs. PRCV-AR310 was isolated from the intestines of a nursery pig from a herd with endemic transmissible gastroenteritis. The AR310 isolate was plaque purified and cell culture propagated, passed once in a gnotobiotic pig, then used as inoculum for a gnotobiotic pig pathogenicity study. Eight pigs were inoculated oronasally with 2 x 10(6) plaque-forming units of PRCV-AR310. Eight pigs served as controls and received cell culture medium. Two pigs from each group were necropsied at 3, 5, 10, and 15 days postinoculation (DPI). There was moderate multifocal to coalescing reddish tan consolidation of 60% of the lung by 10 DPI. Microscopic examination revealed a necrotizing and proliferative bronchointerstitial pneumonia characterized by necrosis, squamous metaplasia, dysplasia, proliferation of airway epithelium, mononuclear cell infiltration of alveolar septa, mild type II pneumocyte proliferation, and lymphohistiocytic alveolar exudation. The microscopic lesions were mild by 3 DPI, moderate by 5 DPI, severe by 10 DPI, and mostly resolved by 15 DPI. No lesions were observed in the intestines of these pigs. There was no clinical respiratory disease. Control pigs remained normal and had no lesions. PRCV was isolated from the lungs but not from the intestines of inoculated pigs. PRCV was not isolated from the lungs or intestines of control pigs. PRCV was also isolated from the nasal and rectal swabs of inoculated but not of control pigs.

Animals↗

Identification, cloning and functional characterization of novel sperm associated antigen 11 (SPAG11) isoforms in the rat.

BACKGROUND: Sperm binding proteins and their C-terminal peptides of the Sperm Associated Antigen 11 (SPAG11) family were found to play an important role in epididymal innate immunity in addition to their role in sperm maturation. However, the expression of Spag11 transcripts in rodents is not well documented. METHODS: Computational analysis was employed to identify novel Spag11 isoforms in the rat. RT-PCR analyses were carried out on RNAs isolated from the male reproductive tract tissues of rat using gene specific primers for Spag11c and Spag11t. The identities of PCR products were confirmed by sequencing. Tissue distribution, developmental expression and androgen regulation of Spag11t and Spag11c were studied using RT-PCR. The antimicrobial activities of recombinant Spag11t and Spag11c were tested against E coli in a colony forming unit assay. RESULTS: In this study, we identified two novel Spag11 transcripts, namely, Spag11t and Spag11c derived from the long arm of chromosome 16 in the rat (Rattus norvegicus), using both in silico and molecular biology approaches. Spag11c is expressed in all three regions of the epididymis, in testis and in ovary but is absent from the seminal vesicle. Spag11t expression is confined to the caput and it is not expressed in the testis, seminal vesicle or ovary. Age dependent expression of Spag11t and Spag11c was observed in the epididymides of rats (10-60 day old). Their expression was found to be most abundant in the adult rat (60 day) suggesting roles in mature reproductive function. Further, both Spag11t and Spag11c expression was down regulated in castrated rat epididymides and the expression was maintained in the testosterone replaced castrated rats. SPAG11C is a potent antibacterial agent. SPAG11T also displayed bactericidal capacity although weaker than SPAG11C and SPAG11E. CONCLUSION: The abundant expression of Spag11t and Spag11c in the male reproductive tract suggests an important role in male reproductive tract immunity. Their expression is developmentally regulated and androgen dependent. Characterization of novel SPAG11 isoforms will contribute to our understanding of the role of epididymal proteins in sperm maturation and innate immunity.

Aging↗

Hormonal evidence for induced ovulation in Monodelphis domestica.

Breeding activity in the South American grey opossum, Monodelphis domestica, is stimulated by the presence of males. This study presents the first analysis of changes in concentrations of plasma progesterone during pregnant and nonpregnant cycles. In Expt 1, females were paired with either intact or vasectomized males to stimulate a reproductive cycle, or were isolated from males. Within 4-8 days of pairing, females showed marked changes in the size of the urogenital opening (vulva), which were paralleled by similar changes in body weight (11.4 +/- 2%; mean +/- SEM; n = 13). There was a second increase and decrease in body weight (21.7 +/- 3.3%; n = 13) in pregnant and nonpregnant cycles during the luteal phase of the cycle. Changes in concentrations of plasma progesterone, determined from samples collected three times a week, were similar in pregnant and nonpregnant cycles. Plasma progesterone concentration, which was increased for 15-18 days, showed two distinct peaks. The first peak, 3-4.5 ng ml-1, occurred immediately after the decrease in vulval swelling, and the second peak (1-2 ng ml-1) occurred 8-12 days later. Concentrations of plasma progesterone were low before parturition occurred. Females isolated from males for 35 days showed no changes in body weight or vulval swelling and concentrations of plasma progesterone remained undetectable. In Expt 2, concentrations of luteinizing hormone (LH) in plasma were determined twice a day for the first few days after pairing. In three of five females an LH peak was detected coincident with the maximal swelling of the vulva.

Animals↗

Rabbit sex hormone binding globulin: primary structure, tissue expression, and structure/function analyses by expression in Escherichia coli.

Sex hormone binding globulin (SHBG) is a homodimeric plasma protein found in mammals that binds sex steroids with high affinity and regulates their bioavailability. The protein is identical in structure and properties to the androgen binding protein (ABP) found in the male reproductive tract. We have isolated a 1245-base pair rabbit SHBG cDNA encoding a reading frame for a signal peptide followed by a protein of 367 amino acids, which shares 79.0, 68.1 and 63.2% amino acid identity with the corresponding human, rat and mouse proteins respectively. Northern blot and hot-nested PCR analyses indicated that rabbit SHBG is produced from a 1.6 kilobase mRNA in the liver of both sexes and in the testis. The rabbit SHBG cDNA was inserted into pGEX-1 lambda T for expression of a glutathione S-transferase/SHBG fusion protein in Escherichia coli. The bacterial product bound 5 alpha-dihydrotestosterone (DHT) in the same manner as the corresponding protein in serum. The dissociation constants (Kd) for rabbit and human SHBGs produced in E. coli were 11.1 +/- 1.1 nM and 2.1 +/- 0.6 nM respectively, and rabbit SHBG formed a less stable protein-steroid complex (t1/2 = 5 min) than human SHBG (t1/2 > 60 min). Unlike human SHBG, rabbit SHBG does not bind estradiol with high affinity. To aid in the identification of differences in the sequences of rabbit and human SHBG, which determine species differences in steroid-binding affinity and specificity, chimeras containing the 5'-terminal half of SHBG from one species and 3'-terminal half of SHBG from the other species were constructed and expressed. It was found that the chimeric proteins assumed similar steroid-binding affinity and specificity as the wild-type proteins when the amino (N)-terminal half of SHBG was derived from the same species. Replacement of the carboxyl (C)-terminal half of rabbit SHBG by the corresponding region of the human molecule increased the integrity of its steroid-protein complex. This supports the concept that amino acids within the N-terminal half of SHBG constitute the steroid-binding domain while the C-terminal half of the molecule may provide structural stability to the protein and its steroid-binding site.

Amino Acid Sequence↗

Identification of the growth hormone receptor in an advanced teleost, the tilapia (Oreochromis mossambicus) with special reference to its distinct expression pattern in the ovary.

There is considerable evidence that the GH/IGF-I axis plays an important role in female reproduction. We report the isolation and characterization of the GH receptor (GH-R) and its gene expression profile during oogenesis in the tilapia, Oreochromis mossambicus. cDNA encoding GH-R was cloned and sequenced from the tilapia liver. The predicted GH-R preprotein consisted of 635 amino acids and contained a putative signal peptide, an extracellular region with a characteristic motif, a single transmembrane region, and a cytoplasmic region with conserved box 1 and 2 domains. The tilapia GH-R shared 34-74% identities with known GH-Rs in vertebrates. A binding assay using COS-7 cells showed that the cloned GH-R bound specifically to tilapia GH. Northern blot analysis showed a single mRNA transcript in the liver and ovary. In situ hybridization revealed intense signals of GH-R in the cytoplasm and nucleus of immature oocytes. The granulosa and theca cells surrounding vitellogenic oocytes also contained the GH-R mRNA signals. About a tenfold greater level of GH-R mRNA was found in the immature oocytes versus the mature oocytes, along with high levels of IGF-I mRNA. There were no significant changes in mRNA levels of GH-R and IGF-I in the liver or in plasma IGF-I levels during oocyte development. No correlation was found between hepatic GH-R mRNA and ovarian GH-R mRNA. These results suggest that the GH/IGF-I axis in the ovary may be involved in the early phases of oogenesis, under a different regulatory mechanism of GH-R gene expression from that of the liver.

Animals↗

Restricted feeding and reproductive performance of individually caged broiler breeder males.

Two experiments were conducted, one in the summer and another in the fall, involving 95 Hubbard broiler breeder males per experiment. Breeder males were randomly placed in individual cages at 26 (Trial 2) or 30 (Trial 1) weeks of age and provided with five levels of feed intake. The Hubbard White Mountain male Management Guide recommendation (154 g/bird/day) was designated as the control level (100%). Other levels were 115, 85, 70, and 55% of the control level. The effects of feed intake on body weight, semen volume, percent semen-packed cell volume, luteinizing hormone (LH), and testes weight were determined. Body weights were significantly decreased as feed intake was decreased. Dietary restriction levels of 115, 100, 85, and 70% had no significant effects on semen volume or semen-packed cell volume. Dietary restriction of 55% resulted in reduced semen volume, semen-packed cell volume (except in Trial 1), and testes weight. Dietary restriction had no significant effect on LH. A calculated metabolizable energy (ME) value of 346 kcal of ME/bird/day, which lies between the 85 and 70%-fed groups, should be adequate for normal body weight maintenance with the Hubbard broiler breeder male grown in individual cages.

Animals↗

Chromosome abnormalities as a cause of infertility in mares.

Chromosomal abnormalities have been detected in seven mares isolated by their poor reproductive performance. All had small or rudimentary gonads and absent or irregular oestrous cycles. Two mares had an XO genotype, one was a 65,XXX female and another a 64,XY sex-reversed female. Two other mares were sex chromosome mosaics of the 63,X/64,XX type. The seventh mare showed a normal female karyotype but a small extra autosomal fragment was found in a few cells.

Animals↗

School based youth health nurses' role in assisting young people access health services in provincial, rural and remote areas of Queensland, Australia.

INTRODUCTION: People living in rural and remote areas have poorer health and more limited access to health services than those in urban areas. Adolescence, generally a period of optimum health, is fraught with risky health behaviours, increasing morbidity and mortality. Increasingly, mental health problems including depression, alcohol and drug use, suicide, self-harm, sexual health, inappropriate nutrition and physical activity are key areas of adolescent health concern in Queensland, Australia. To address the health and well being needs of young people the School Based Youth Health Nurse (SBYHN) Program was introduced into State high schools in Queensland in 1998. SBYHN responsibilities include individual consultations and health promotion within the high school environment. This study was undertaken to explore the referral role of SBYHN in provincial, rural and remote Queensland and to ascertain the availability of referral services. METHOD: A self-report survey was posted to all SBYHN in Queensland. Forty-four (62%) nurses participated; 27 from provincial, 11 from rural and 6 from remote areas. RESULTS: SBYHN consult with young people in provincial, rural and remote State high schools in the areas of psychological concerns, drug and alcohol concerns and/or abuse, family conflict and/or abuse, sexual and reproductive health, social and isolation concerns and nutritional advice. However, the frequency with which SBYHN reported themselves as an appropriate source for young people to seek assistance from varied from over 90% for areas such as sexual and nutritional advice to approximately 30% for areas such as physical abuse and drug and alcohol abuse. When required by the SBYHN, referral services were locally accessible to young people less than 50% of the time. Access to some referral services was reduced because some young people do not want parental involvement in their health and the service requires parental consent. When referral services were unavailable SBYHN used phone counselling, available generalists, supported and monitored the young people until a service was available, or they accepted this as an aspect of residing outside the metropolitan areas and did the best they could. Sometimes, when a young person was reluctant to attend a recommended service or there was a long wait for an appointment, SBYHN provided ongoing support while endeavouring to persuade attendance at the recommended service. Situations like these, experienced by at least half those studied, place SBYHN in the precarious position of practicing beyond their scope of practice. CONCLUSIONS: Health problems during adolescence can be reduced by ensuring assistance and support are available for young people when and if they require it. Thorough confidential individual consultations are provided by SBYHN and young people are availing themselves of this service. SBYHN are practicing at an advanced level and need to be recognised as such. Additionally, the availability of services in rural and remote regions needs consideration. As rural youth are more vulnerable to a number of health concerns, it is imperative that services are available for them. The issue of parental consent remains a barrier to young people seeking health services and it may be timely for community debate on this issue.

Journal Article↗

X-ray analysis of biological fluids: contribution of microdroplet technique to biology.

Electron probe analysis by x-ray spectrometry is used in biology for simultaneous determination of the concentrations of any elements with a higher atomic number than that of carbon in single samples with volumes of 0.01 to 0.5 nl. In this technique, deposits prepared from identical volumes of biological fluids and standard solutions are totally covered by the electron beam, and the measured x-ray intensities for each element directly compared. The possibility of intensity quantification depends on the thinness of the dried deposits obtainable by various preparatory techniques. Factors affecting the accuracy of the results include droplet stability under the electron beam, the identity of the degree of oxidation of the elements in biological fluids and standards, sample mass thickness, beam voltage, and matrix effects. Minimum detectable concentrations in the 0.05 mmol.l-1 range are now achievable. This technique is the only one applicable in cases where available volumes are too small to determine the concentrations of several elements on the same sample (for instance of Na, Mg, S, P, Cl, K, Ca, Fe and Co), or even to determine the concentration of a single element (e.g. Mg). Although the droplet technique has so far mainly been used in renal physiology, it has also been applied in reproductive and digestive physiology. Isolated cells are analyzed according to the same principle of totally covering the cell by the electron beam. During the last decade, the vast increase in the relevant literature has testified to the contribution of the microdroplet technique to various fields of biology.

Absorption↗

[The antilysozymal activity and species characteristics of the microflora of the female reproductive tract during pregnancy].

The results of the study of the species composition and the antilysozyme activity of the microflora the reproductive tract of pregnant women are presented. Changes in the microflora of the reproductive system of pregnant women have been registered. These changes are characterized by the increased proportion of staphylococci in biotopic samples of the reproductive tract and the isolation of enterobacterial strains with high antilysozyme activity.

Adult↗

Isolation, characterization, and radioimmunoassay of Atlantic halibut somatolactin and plasma levels during stress and reproduction in flatfish.

Somatolactin (SL), a recently identified teleost pituitary hormone which is a member of the growth hormone/prolactin family, was isolated from pituitary tissue of Atlantic halibut (Hippoglossus hippoglossus). Pituitary proteins were extracted in ammonium bicarbonate (pH 7.8), fractionated using gel filtration chromatography, and purified using reversed-phase high-performance liquid chromatography. Halibut SL was identified on the basis of molecular size (determined by gel electrophoresis and mass spectroscopy), cross-reactivity of the putative hormone with antisera to cod SL, and N-terminal amino acid sequence. Polyclonal antibodies to purified halibut SL were raised in rabbits, and a radioimmunoassay (RIA) was developed for measurement of plasma concentrations of SL using purified halibut SL as a standard. The RIA was tested in several flatfish species including Pacific halibut (Hippoglossus stenolepis), English sole (Pleuronectes vetulus), and rock sole (Lepidopsetta bilineata). The assay was specific for SL as indicated by absence of cross-reactivity with Atlantic halibut growth hormone, prolactin, and GTH alpha subunit. Dilutions of plasma and pituitary extracts from Pacific halibut, English sole, and rock sole were parallel to the Atlantic halibut SL standard curve, indicating that the assay is valid for a range of flatfish species. Using halibut SL antiserum, SL was localized in the pars intermedia of English sole pituitary, where it has been identified in previously examined teleost species. The RIA was used to measure plasma levels of SL in Atlantic halibut and English sole during reproductive development, and in English sole subjected to various types of environmental stressors, including handling and crowding. In both sole and halibut, plasma SL concentrations remained relatively constant throughout gonadal development, but dropped during or following ovulation. Plasma SL levels in English sole tended to increase in response to acute stress, in parallel with plasma cortisol levels.

Amino Acid Sequence↗

Porcine leptospirosis in Iowa.

The epidemiology of leptospirosis in Iowa swine was examined on the basis of serologic results and herd data from 55 herds in the National Animal Health Monitoring System (NAHMS) program and culture results and histories from 578 cases of reproductive failure submitted to the Iowa Veterinary Diagnostic Laboratory during a 3-year period. Thirty-eight percent of sera from NAHMS herds contained antibodies against 1 or more of 12 leptospira antigens. Leptospires were isolated from 9 (1.6%) of 578 cases of reproductive failure. Seven (78%) of the isolates were identified as Leptospira interrogans serovar kennewicki and 2 (22%) as serovar grippotyphosa. In 7 herds from which leptospires were isolated, attack rates ranged from 1% to 84%. Clinical leptospirosis, characterized by reproductive failure and confirmed by isolation of leptospires, was sporadic. No significant differences in farrowing averages and reproductive problems were observed between vaccinated and nonvaccinated NAHMS herds or between herds with higher (43-63%) or lower (14-40%) percentages of animals that were serologically positive against serovar bratislava.

Abortion, Veterinary↗