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Effects of midecamycin acetate (miocamycin), a new macrolide antibiotic, on reproductive performances in rats and rabbits.

Miocamycin (MOM) is a derivative of midecamycin, a macrolide antibiotic isolated from a culture broth of Streptomyces mycarofaciens. The objective of this study was to determine the effect of MOM on reproductive performance in rats and rabbits. MOM, non-crystalline solid, was suspended in 0.1% CMC solution immediately before use. In this study, reproductive performance was studied according to the following designs: Fertility test (Segment I) in Wistar rats Teratogenicity test (Segment II) in Wistar rats Peri- and post-natal tests (Segment III) in Wistar rats Behavioral test in rat newborns in Segments II and III Teratogenicity test (Segment II) in New Zealand white rabbits In conclusion, MOM, non-crystalline solid, might have no teratogenesis and little influence on dams and their fetuses and offsprings.

Abnormalities, Drug-Induced↗

Role, mechanism of action and application of gonadoliberins in reproductive processes.

Gonadoliberin (gonadotropin releasing hormone, GnRH) plays a central role in the regulation of reproductive functions as it regulates the release of both luteinizing hormone (LH) and follicle stimulating hormone (FSH). The isolation and structure determination of GnRH opened the possibility of its use for influencing reproductive processes. This possibility initiated a rapid development in the design of potent and long-acting GnRH agonists and antagonists. The most important structural modifications of GnRH leading to superagonists are the D-amino acid substitutions in position 6 combined with Pro9-ethylamide or azaGly10 at the C-terminus. We have synthesized several superagonists of GnRH according to these substitution principles. Furthermore, our L-isoaspartyl modification in position 6, as a new approach to GnRH agonist design, also resulted in superactive analogs. The recently discovered sequences of non-mammalian GnRH-s opened new routes for us to synthesize species specific GnRH agonists. All three groups of the above mentioned GnRH analogs have been successfully used for the treatment of sexual disorders of different animals (cattle, pigs, rabbits, etc.). Ovulation synchronization and a 30% increase in the fertility rate could be achieved by using GnRH agonists in cattle breeding. Analogs derived from species specific sequences could be applied for the induced artificial propagation of fish even out of the spawning season. It is known that superactive GnRH analogs can suppress the growth of certain hormone-dependent tumours. In vitro and in vivo tests of our analogs showed promising antitumour activity in breast cancer which might be explained by the mechanism of desensitization. Almost a hundred antagonist analogs of GnRH have been developed in our laboratory. The most effective ones contain 4 or 5 D-amino acids, and one of them is even orally active. The inhibition of ovulation can also be achieved by the administration of GnRH superagonists. This phenomenon might also be explained by the desensitization of LH-release. Radioactive analogs specifically labeled with tritium in different amino acid residues have been synthesized and used for studying tissue distribution and biodegradation of gonadoliberins. Analogs containing a photoreactive group have been prepared and applied for the trials of GnRH receptor isolation.

Amino Acid Sequence↗

[In vitro patogenicity of the fungi Metarhizium anisopliae (Metschnikoff, 1879) Sorokin, 1883, on the tick Boophilus microplus (Canestrini,1887)].

The cattle tick Boophilus microplus is a serious concern to the Brazilian cattle industry. It causes decreased meat and milk production, low alimentary conversion, damage to leather and transmission of pathogens. The use of entomopathogenic fungi as biological control agents for ticks has shown promising results. This study tests the in vitro effects of two Metarhizium anisopliae isolates (E9 and 319) towards three stages of B. microplus: eggs, larvae and engorged females. The bioassays were composed of five treatment groups (concentrations: 10(5), 10(6), 10(7), 10(8) conidia/ml and the control) each one with ten repetitions. The stages were treated by immersion for three minutes. After the treatments, the females, eggs and larvae were incubated at 27 degrees C +/- 1 and RH >or= 80%. The main studied parameters were: percentage of larvae eclosion, percentage of larvae mortality and the indexes of nutritional and reproductive efficiency. In vitro tests of the fungal isolates showed effective control of the three stages of B. microplus, suggesting high potential for their use as a microbial control agent of the B. microplus tick.

Animals↗

Premature ovarian failure (POF): discordance between somatic and reproductive aging.

Premature ovarian failure (POF) is a unique example of isolated organ senescence, with a population prevalence of approximately 1%. Though the phenotypic expression of POF is similar to that of age-appropriate natural menopause, the underlying pathophysiological mechanisms are diverse and not entirely clear. The impact of POF on the patient is profound, with myriad ramifications, ranging from psychological devastation to multi-system implications of estrogen deprivation and its sequelae. The hastening of degenerative changes noted in these patients however, are not entirely ameliorated with estrogen replacement and POF may indeed represent an acceleration of the aging process.

Aging↗

Increased PGE2 from human monocytes isolated in the luteal phase of the menstrual cycle. Implications for immunity?

The reproductive hormones are implicated in the well documented sexual dimorphism in cellular and immune responses. Prostaglandins (PGs) are mediators of the immune response with their concentration and relative amounts being pivotal to their impact. In resident peritoneal macrophages isolated from mice we had previously noted that the cells from females synthesized significantly more PG than males. In these experiments we investigated whether PG metabolism in the human monocyte was influenced by gender and by stage of the menstrual cycle. Monocytes isolated from the female and activated in vitro with LPS produced on average significantly more PG into the medium than the males. Among females, significantly more PG was found in the medium from cells isolated during the luteal phase of the cycle than during the early follicular phase. It was also in this luteal phase in which the female differed substantially from males. We suggest that the in vivo hormonal changes associated with the menstrual cycle modulate monocyte synthesis of PG and other immune modulators such as IL-1. This could be a key to understanding differences in vulnerability between males and females as well as within phases of the cycle, to immune and inflammatory insult.

Adult↗

[Testosterone-7alpha-3H uptake by the anlage of organs related to reproduction in female rate embryos in experiments in vivo and in vitro].

The dynamics of testosterone-7alpha-3H (T-3H) uptake by the developing genital system organs, as well as by the suprarenals, brain cortex and muscles was studied in vivo experiments with the 14-20 days old female rat embryos. Besides, the T3H uptake by the reproductive tract tissues and muscles just after the isolation of embryos from the uterus and after 2 days cultivation in the diffusion chambers implanted in the abdominal cavity of the adult castrated rats was studied in in vitro experiments. Moreover, the T3H uptake by the embryonic tissues was determined in the absence of ovaries. The specific uptake was noted in all the tissues at all developmental stages under all experimental conditions. The selective T3H uptake was noted during the periods corresponding to the rudiments' morphogenesis carachteristic for males. In the absence of ovaries, the T3H uptake by the reproductive tract tissues in vitro experiments decreased, but after 2 days cultivation in the diffusion chambers increased. The possible participation of ovaries in the female sex differentiation and the bisexual nature of rudiments determined by the presence of receptors both to estro- and androgens are discussed.

Animals↗

A gene encoding proline dehydrogenase is not only induced by proline and hypoosmolarity, but is also developmentally regulated in the reproductive organs of Arabidopsis.

The cDNA clone ERD5 (early responsive to dehydration), isolated from 1-h-dehydrated Arabidopsis, encodes a precursor of proline (Pro) dehydrogenase (ProDH), which is a mitochondrial enzyme involved in the first step of the conversion of Pro to glutamic acid. The transcript of the erd5 (ProDH) gene was undetectable when plants were dehydrated, but large amounts of transcript accumulated when plants were subsequently rehydrated. Accumulation of the transcript was also observed in plants that had been incubated under hypoosmotic conditions in media that contained L- or D-Pro. We isolated a 1.4-kb DNA fragment of the putative promoter region of the ProDH gene. The beta-glucuronidase (GUS) reporter gene driven by the 1.4-kb ProDH promoter was induced not only by rehydration but also by hypoosmolarity and L- and D-Pro at significant levels in transgenic Arabidopsis plants. The promoter of the ProDH gene directs strong GUS activity in reproductive organs such as pollen and pistils and in the seeds of the transgenic plants. GUS activity was detected in vegetative tissues such as veins of leaves and root tips when the transgenic plants were exposed to hypoosmolarity and Pro solutions. GUS activity increased during germination of the transgenic plants under hypoosmolarity. The relationship between Pro metabolism and the physiological aspects of stress response and development are discussed.

Arabidopsis↗

Roles of inhibins, activins, and follistatin in the female reproductive system.

Some 10 years have elapsed since inhibins were first isolated from ovarian follicular fluid and characterized as disulphide-linked dimeric glycoproteins capable of selectively suppressing the synthesis and secretion of follicle-stimulating hormone (FSH) by pituitary gonadotropes. There have been numerous surprises subsequent to this break-through, including the discovery and molecular characterization of activins and follistatins, proteins which share with inhibin an ability to modulate pituitary FSH secretion. It has also emerged that (i) inhibins and activins are members of the diverse but structurally related transforming growth factor beta (TGF beta) peptide family; (ii) follistatin, although structurally unrelated to the TGF beta family, modulates activin bioactivity by acting as a specific high-affinity binding protein; (iii) inhibins, activins, and follistatin function as intragonadal autocrine/paracrine regulators of follicle cell differentiation and steroidogenesis; (iv) inhibins, activins, and follistatin are expressed and subserve local regulatory roles in numerous extragonadal tissues, including brain, adrenal, bone marrow, and placenta but perhaps most notably in anterior pituitary--the classical target tissue for inhibin; (v) the activin-follistatin system may play a key role in early embryogenesis. This article focuses on these developments of the past decade with particular reference to the distribution and functional roles of inhibins, activin, and follistatin in tissues related to the female reproductive system, including hypothalamus, anterior pituitary, ovary, and placenta.

Activins↗

Analysis of endophyte toxins: fescue and other grasses toxic to livestock.

Research on livestock toxicoses caused by Acremonium (endophyte)-infected grasses strongly implicate the ergopeptine alkaloids with A. coenophialum-infected fescue and paxilline and the lolitrem alkaloids with A. lolii-infected perennial ryegrass as the causative agents. Isolation, identification, and detection of these toxins involves extraction with appropriate solvents, clean-up procedures, and chromatographic methods with known standards. Thin-layer, high-performance liquid and gas chromatography along with ultraviolet and mass spectrometric (i.e., electron impact, chemical ionization, tandem mass) characterizations have been reported. These methods have varying degrees of success depending on the matrix from which the alkaloids have been extracted. Ergovaline is the primary ergopeptine alkaloid isolated from cultures of A. coenophialum and also from infected fescue grass and seeds toxic to livestock. Other compounds isolated from the endophyte-infected fescue include: lysergic acid amide (ergine), the clavine class of ergot alkaloids (chanoclavine I, agroclavine, elymoclavine, penniclavine), the pyrrolizidine alkaloids (N-formylloline, N-acetylloline, N-methyloline, N-acetylnorloline), and the unique pyrrolopyrazine alkaloid peramine. The loline alkaloids and peramine have been more associated with the insect-deterrent properties of the endophyte-infected fescue than with livestock toxicoses. Also, both peramine and the ergopeptine alkaloids (ergovaline, ergotamine) have been isolated from A. lolii-infected perennial ryegrass. More recently, paxilline and lolitrem B have been detected in laboratory cultures of A. coenophialum isolated from tall fescue. The ergot alkaloids in endophyte-infected perennial ryegrass may be more related to decreased animal productivity (weight gains, reproduction problems), whereas the lolitrems cause the staggers syndrome. The detection, isolation, identification, and analyses of these compounds from Acremonium-infected grasses is presented.

Acremonium↗

Effects of allogeneic contact on life-history traits of the colonial ascidian Botryllus schlosseri in Monterey Bay.

The formation of chimeric colonies following allogeneic contact between benthic invertebrates may strongly affect colony fitness. Here we show that, in a field population of the colonial ascidian Botryllus schlosseri in Monterey Bay, California, more than 20% of all colonies occur in allogeneic contact with conspecifics. We experimentally assessed the effects of allogeneic contact on the following life-history traits under natural field conditions: growth, age and size at first reproduction, and egg production (fecundity). When compared with isolated colonies, and in some cohorts also with colonies that rejected allogeneic neighbors, colonies that fused with neighbors incurred reduced fitness in terms of most life-history traits measured. We propose that one of the benefits of precise allorecognition is that, in fused colonies, it limits the unit of selection to chimeric individuals composed of closely related kin.

Animals↗

Genetic variability of encephalomyocarditis virus (EMCV) isolates.

In order to evaluate the variability of encephalomyocarditis virus (EMCV), field isolates originating from different European regions and inducing different clinical pictures in pigs have been molecularly characterised. The regions targeted were the poly(C) tract, a part of the 5'-UTR (360 nucleotides), the Leader gene (201 nucleotides), the complete capsid coding region (2502 nucleotides), the 2A gene (403 nucleotides), the end of the 3D polymerase gene (305 nucleotides) and the 3'-UTR (123 nucleotides). Analyses have also been performed on a virulent field isolate, which had been subjected to serial passages in vivo and in vitro resulting, in the case of the in vitro passaged virus, in attenuation, as demonstrated by animal experiments. The present study shows that different clinical pictures, such as acute fatal myocarditis or reproductive failure, may not only be caused by EMCV isolates which are genetically diverse but also by the same isolate. Thus no correlation could be demonstrated between genotype and clinical disease. However, the European isolate which showed the highest genetic divergence also gave rise to a more complex clinical picture. Despite EMCV having been isolated from cases of acute fatal myocarditis in pigs in certain areas of the world for many years, clinical disease, including a variety of clinical pictures and pathogenicity, has only been recognised in Europe since 1986 and thus it can be considered an emerging disease in this region. These findings, associated with the reported phenotype changes of the virus under environmental changes (passages), along with its wide distribution among vertebrate species (including higher primates), shows the validity of considering EMCV as a potential pathogen for recipients in xenotransplantation.

3' Untranslated Regions↗

Reproduction trends and the emergence of moral panic.

Are we witnessing a moral panic or a moral evaluation in the responses to the new reproductive technologies? Locating women in reproductive technology is a particularly powerful way into a discussion of values and consequences, intended or unintended. Twelve areas are isolated for discussion beginning with issues of definition and location: What is reproductive and genetic engineering? and How does human reproduction fit into bio-technological developments generally? Next issues are raised about how the new developments are being shaped ideologically and how they are being promoted by the state. A series of legal issues affecting women, men, children, embryos and fetuses follow. The focus then shifts to the way women are ideologically constructed: Why is there an assumption that woman = mother, and mother = heterosexual married or stable cohabiting woman?--and socially controlled: Why assume technological change in human reproductive processes means the basic subordination of women to men will not alter? The usefulness of the new technologies in relieving involuntary childlessness is then queried which leads on to a questioning of whether these developments are qualitatively different from other technologies used in human reproduction. Futuristic concerns such as the importance of the male pregnancy and the social consequences of fully medicalising human reproduction are raised. The last area to be considered is protest by women and suggested alternative approaches to increase the social visibility and recognition of the needs and interest of women in the current and potential use of the new medical and scientific developments.

Female↗

Occurrence of deformed fish and their fitness-related traits in Kirikuchi charr, Salvelinus leucomaenis japonicus, the southernmost population of the genus Salvelinus.

Kirikuchi charr, Salvelinus leucomaenis japonicus, is the southernmost population of the genus Salvelinus. It is endemic to the Kii Peninsula, central Honshu Island, Japan. As a consequence of anthropogenic disturbances, a few populations of Kirikuchi charr with low genetic diversity now survive only in small, isolated habitats. This study investigated the occurrence of deformed individuals and assessed differences between deformed and nondeformed fish in fitness-related traits, i.e., body size, body condition, growth rate, reproductive traits, survival rate, and habitat use, for two small isolated populations of Kirikuchi charr in the upper drainage of the Totsu River system of the Kii Peninsula. The two populations contained deformed fish in the respective proportions of 8.0-17.4% and 3.0-5.8% between 2003 and 2005. Annual survival rates of deformed fish were approximately half those of nondeformed fish. Other traits were not significantly different between deformed and nondeformed fish. These results indicate that the occurrence of deformities is an important ecological indicator that reflects the decline in fitness of small, isolated populations.

Adaptation, Physiological↗

An investigation of male-mediated F1 effects in mice treated acutely and sub-chronically with urethane.

In order to investigate the alleged potential of paternally administered urethane to cause foetal abnormalities and heritable tumours, male CD-1 mice were treated with urethane, either acutely by intraperitoneal injection at doses of 1.25 and 1.75 g/kg bodyweight (bwt) or sub-chronically in the drinking water at 1.25 for 10 weeks, and 3.75 mg/ml for 9 weeks or vehicle for the control groups. They were mated to untreated females 1 week later. Uterine contents of half the pregnant females were examined just before full term, while the remaining females were allowed to deliver their litters. The resulting F1 offspring were observed for approximately 18 months and 12 months for acute and sub-chronic exposures respectively and subjected to necropsy examination. Some of the mice treated acutely with 1.75 g/kg bwt exhibited partial infertility but none of those treated with 1.25 g/kg bwt had an adverse effect on their reproductive ability. There was no genetic effect of acute urethane treatment on male germ cells as indicated by dominant lethality. After birth, there was an increase (P < 0.05) in post-implantation deaths possibly due to perinatal mortality. There was an increased incidence and earlier onset of liver tumours induced in F1 male offspring from F0 males treated with 1.75 g/kg bwt, (20.7% vs. 10.1%, P = 0.026) but not in the female offspring. F1 males from both treatment groups had mean bodyweights significantly higher than controls (P < 0.01). Some males from each dose group of the acute study were examined using the restriction site mutation assay involving analysis of exon sequences. No mutations were identified in testes, liver or spleen of DNA isolated from the urethane-treated animals. No reproductive or genetic effects were seen with sub-chronic treatment at either 1.25 or 3.75 mg/ml urethane nor was there any predisposition of F1 animals to tumours although observation times were shorter.

Abnormalities, Drug-Induced↗

Hydrogen peroxide producing lactobacilli in women with vaginal infections.

OBJECTIVE: Hydrogen peroxide producing lactobacilli are isolated from the vaginas of a majority of healthy reproductive age women. Their toxic and inhibitory effect against the overgrowth of pathogens in the vagina is documented by in vitro studies. Clinical studies concerning the role of hydrogen peroxide producing lactobacilli in vaginal infections are controversial. STUDY DESIGN: The aim of this study was to isolate lactobacilli from women with major vaginal infections: bacterial vaginosis (BV), vulvovaginal candidiasis (VVC) and trichomoniasis (TV), and from women with normal flora (NF), to test their ability to produce hydrogen peroxide, to quantitate lactobacilli and to evaluate factors related to the colonization of vagina with hydrogen peroxide producing lactobacilli. The study group comprised 108 women: 27 with VVC, 26 with BV, 20 with TV and 35 with NF. Data was analysed with Fisher exact test, chi2-test, and ANOVA. RESULTS: Lactobacilli were isolated from 81.5% of women with VVC, 58% of women with BV (p=0019), 55% of women with TV (p=0.015) and from 86% of women with NF. The majority of isolates of lactobacilli from women with VVC, TV and NF produced hydrogen peroxide (77%, 63.5% and 80%, respectively), compared with women with BV where only 26.5% of isolates produced hydrogen peroxide (p=0.01). Quantitative analysis showed that in an overwhelming majority of women colonized with lactobacilli, their count was >10(7) CFU/ml. Upon evaluation of behavioural and demographic characteristics, we found that smokers had a lower incidence of isolation of hydrogen peroxide producing lactobacilli (p=0.018). Our results showed that women with BV lack lactobacilli, especially hydrogen peroxide producing ones. Women with TV had a lower rate of lactobacilli. In women with VVC, there was neither significant difference in the isolation of lactobacilli, nor in their ability to produce hydrogen peroxide, compared to women with NF. CONCLUSION: Our results point out that hydrogen peroxide producing lactobacilli could protect against the development of BV, but not against VVC and TV.

Adolescent↗

Cloning and characterization of the mating type (MAT) locus from Ascochyta rabiei (teleomorph: Didymella rabiei) and a MAT phylogeny of legume-associated Ascochyta spp.

Degenerate primers designed to correspond to conserved regions of the high mobility group (HMG) protein encoded by the MAT1-2 gene of Cochliobolus heterostrophus, Cochliobolus sativus, and Alternaria alternata were used to amplify the portion of the sequence corresponding to the HMG box motif from Ascochyta rabiei (teleomorph: Didymella rabiei). A combination of TAIL and inverse PCR extended the MAT1-2 sequence in both directions, then primers designed to MAT1-2 flanking DNA were used to amplify the entire MAT1-1 idiomorph. MAT1-1 and MAT1-2 idiomorphs were 2294 and 2693 bp in length, respectively, and each contained a single putative open reading frame (ORF) and intron similar to MAT loci of other loculoascomycete fungi. MAT genes were expressed at high levels in rich medium. MAT-specific PCR primers were designed for use in a multiplex PCR assay and MAT-specific PCR amplicons correlated perfectly to mating phenotype of 35 ascospore progeny from a cross of MAT1-1 by MAT1-2 isolates and to the mating phenotype of field-collected isolates from diverse geographic locations. MAT-specific PCR was used to rapidly determine the mating type of isolates of A. rabiei sampled from chickpea fields in the US Pacific Northwest. Mating type ratios were not significantly different from 1:1 among isolates sampled from two commercial chickpea fields consistent with the hypothesis that these A. rabiei populations were randomly mating. The mating type ratio among isolates sampled from an experimental chickpea field where asexual reproduction was enforced differed significantly from 1:1. A phylogeny estimated among legume-associated Ascochyta spp. and related loculoascocmycete fungi using sequence data from the nuclear ribosomal internal transcribed spacer (ITS) demonstrated the monophyly of Ascochyta/Didymella spp. associated with legumes but was insufficiently variable to differentiate isolates associated with different legume hosts. In contrast, sequences of the HMG region of MAT1-2 were substantially more variable, revealing seven well-supported clades that correlated to host of isolation. A. rabiei on chickpea is phylogenetically distant from other legume-associated Ascochyta spp. and the specific status of A. rabiei, A. lentis, A. pisi, and A. fabae was confirmed by the HMG phylogeny

Amino Acid Sequence↗

Expressed sequence tags of fruits, peels, and carpels and analysis of mRNA expression levels of the tagged cDNAs of fruits from the Fuji apple.

In order to understand molecular events during fruit development and provide genetic resources for molecular breeding, 430 expressed sequence tags (ESTs) were generated from randomly selected clones of cDNA libraries prepared from young fruits, peels of mature fruits, and carpels of the Fuji apple (Malus domestica Borkh.). Database comparisons of the ESTs revealed that 180 non-redundant clones showed a high similarity with previously identified genes. Among these, 138 clones exhibited a homology with previously identified plant genes and 12 were identical to genes that were previously identified from apples. The deduced amino acid sequences of 42 clones had a homology to proteins that have not been reported from plants. Eighteen cDNA clones from the young fruit library were selected for studying expression levels and patterns in reproductive organs and leaves. This study revealed that the clones can be classified into 3 different groups based on their expression levels. The first 9 clones were expressed strongly in at least one reproductive organ. Eight of these clones (vacuolar processing protease, sucrose phosphate synthase, arabinogalactan protein, UDP-glucose glucosyl transferase, major allergen D1, cystein proteinase inhibitor, lipoxygenase, and protease subunit SUG2) were highly expressed in mature flowers and young fruits, whereas one clone (z-carotene desaturase protein precursor) was preferentially expressed in mature flowers but weakly in young fruits. The second group includes 6 cDNA clones (glucose transport protein, aminomethyl transferase precursor protein, dTDP-D-glucose-4,6-dehydrogenase, 2 types of protein kinase, and selenium binding protein) that were weakly expressed. These clones were characterized by their preferential expression patterns in mature flowers and young fruits. The transcripts of 3 cDNA clones in the third group (vacuolar aminopetidase, beta-galactosidase, and EREBP-4) were detectable only by RT-PCR and they were preferentially expressed in young fruits. These results indicate that most ESTs that were isolated from young fruits are preferentially expressed in reproductive organs and thereby play important roles during reproductive organ development.

DNA, Complementary↗

Evidence of a uteroglobin-like protein in epithelial cells of reproductive and non-reproductive tissues of the rabbit.

An antiserum raised in a goat to a uteroglobin-like protein isolated from uterine fluid of oestrous rabbits was used in an immune fluorescence test to localize an antigen present in the reproductive tract of oestrous and pseudopregnant rabbits and mammary gland tissue. The antigen was also present in the vas deferens and seminal vesicle, but not in testis. Non-reproductive tissues, such as lung, small intestine, bladder and thyroid showed specific fluorescent staining which was eliminated or significantly reduced by absorption of the antiserum with a purified uteroglobin preparation.

Animals↗