KRAS variation and risk of endometriosis

Endometriosis is a common gynaecological disease with symptoms of pelvic pain and infertility which affects 7-10% of women in their reproductive years. Activation of an oncogenic allele of Kirsten rat sarcoma viral oncogene homologue (KRAS) in the reproductive tract of mice resulted in the development of endometriosis. We hypothesized that variation in KRAS may influence risk of endometriosis in humans. Thirty tagSNPs spanning a region of 60.7 kb across the KRAS locus were genotyped using iPLEX chemistry on a MALDI-TOF MassARRAY platform in 959 endometriosis cases and 959 unrelated controls, and data were analysed for association with endometriosis. Genotypes were obtained for most individuals with a mean completion rate of 99.1%. We identified six haplotype blocks across the KRAS locus in our sample. There were no significant differences between cases and controls in the frequencies of individual single-nucleotide polymorphisms (SNPs) or haplotypes. We also developed a rapid method to screen for 11 common KRAS and BRAF mutations on the Sequenom MassARRAY system. The assay detected all mutations previously identified by direct sequencing in a panel of positive controls. No germline variants for KRAS or BRAF were detected. Our results demonstrate that any risk of endometriosis in women because of common variation in KRAS must be very small.

Experimental demonstration of possible cryptic female choice on male tsetse fly genitalia

artículo -- Universidad de Costa Rica. Escuela de Biología, 2009. Este documento es privado debido a restricciones de derechos de autor del publicador.

Reproducing Women: Female Bodily Narratives and English Patriarchies, 1600-1660

During the early Stuart period, England’s return to male monarchal rule resulted in the emergence of a political analogy that understood the authority of the monarch to be rooted in the “natural” authority of the father; consequently, the mother’s authoritative role within the family was repressed. As the literature of the period recognized, however, there would be no family unit for the father to lead without the words and bodies of women to make narratives of dynasty and legitimacy possible. Early modern discourse reveals that the reproductive roles of men and women, and the social hierarchies that grow out of them, are as much a matter of human design as of divine or natural law. Moreover, despite the attempts of James I and Charles I to strengthen royal patriarchal authority, the role of the monarch was repeatedly challenged on stage and in print even prior to the British Civil Wars and the 1649 beheading of Charles I. Texts produced at moments of political crisis reveal how women could uphold the legitimacy of familial and political hierarchies, but they also disclose patriarchy’s limits by representing “natural” male authority as depending in part on women’s discursive control over their bodies. Due to the epistemological instability of the female reproductive body, women play a privileged interpretive role in constructing patriarchal identities. The dearth of definitive knowledge about the female body during this period, and the consequent inability to fix or stabilize somatic meaning, led to the proliferation of differing, and frequently contradictory, depictions of women’s bodies. The female body became a site of contested meaning in early modern discourse, with men and women struggling for dominance, and competitors so diverse as to include kings, midwives, scholars of anatomy, and female religious sectarians. Essentially, this competition came down to a question of where to locate somatic meaning: In the opaque, uncertain bodies of women? In women’s equally uncertain and unreliable words? In the often contradictory claims of various male-authored medical treatises? In the whispered conversations that took place between women behind the closed doors of birthing rooms? My dissertation traces this representational instability through plays by William Shakespeare, John Ford, Thomas Middleton, and William Rowley, as well as in monstrous birth pamphlets, medical treatises, legal documents, histories, satires, and ballads. In these texts, the stories women tell about and through their bodies challenge and often supersede male epistemological control. These stories, which I term female bodily narratives, allow women to participate in defining patriarchal authority at the levels of both the family and the state. After laying out these controversies and instabilities surrounding early modern women’s bodies in my first chapter, my remaining chapters analyze the impact of women’s words on four distinct but overlapping reproductive issues: virginity, pregnancy, birthing room rituals, and paternity. In chapters 2 and 3, I reveal how women construct the inner, unseen “truths” of their reproductive bodies through speech and performance, and in doing so challenge the traditional forms of male authority that depend on these very constructions for coherence. Chapter 2 analyzes virginity in Thomas Middleton and William Rowley’s play The Changeling (1622) and in texts documenting the 1613 Essex divorce, during which Frances Howard, like Beatrice-Joanna in the play, was required to undergo a virginity test. These texts demonstrate that a woman’s ability to feign virginity could allow her to undermine patriarchal authority within the family and the state, even as they reveal how men relied on women to represent their reproductive bodies in socially stabilizing ways. During the British Civil Wars and Interregnum (1642-1660), Parliamentary writers used Howard as an example of how the unruly words and bodies of women could disrupt and transform state politics by influencing court faction; in doing so, they also revealed how female bodily narratives could help recast political historiography. In chapter 3, I investigate depictions of pregnancy in John Ford’s tragedy, ‘Tis Pity She’s a Whore (1633) and in early modern medical treatises from 1604 to 1651. Although medical texts claim to convey definitive knowledge about the female reproductive body, in actuality male knowledge frequently hinged on the ways women chose to interpret the unstable physical indicators of pregnancy. In Ford’s play, Annabella and Putana take advantage of male ignorance in order to conceal Annabella’s incestuous, illegitimate pregnancy from her father and husband, thus raising fears about women’s ability to misrepresent their bodies. Since medical treatises often frame the conception of healthy, legitimate offspring as a matter of national importance, women’s ability to conceal or even terminate their pregnancies could weaken both the patriarchal family and the patriarchal state that the family helped found. Chapters 4 and 5 broaden the socio-political ramifications of women’s words and bodies by demonstrating how female bodily narratives are required to establish paternity and legitimacy, and thus help shape patriarchal authority at multiple social levels. In chapter 4, I study representations of birthing room gossip in Thomas Middleton’s play, A Chaste Maid in Cheapside (1613), and in three Mistris Parliament pamphlets (1648) that satirize parliamentary power. Across these texts, women’s birthing room “gossip” comments on and critiques such issues as men’s behavior towards their wives and children, the proper use of household funds, the finer points of religious ritual, and even the limits of the authority of the monarch. The collective speech of the female-dominated birthing room thus proves central not only to attributing paternity to particular men, but also to the consequent definition and establishment of the political, socio-economic, and domestic roles of patriarchy. Chapter 5 examines anxieties about paternity in William Shakespeare’s The Winter’s Tale (1611) and in early modern monstrous birth pamphlets from 1600 to 1647, in which children born with congenital deformities are explained as God’s punishment for the sexual, religious, and/or political transgressions of their parents or communities. Both the play and the pamphlets explore the formative/deformative power of women’s words and bodies over their offspring, a power that could obscure a father’s connection to his children. However, although the pamphlets attempt to contain and discipline women’s unruly words and bodies with the force of male authority, the play reveals the dangers of male tyranny and the crucial role of maternal authority in reproducing and authenticating dynastic continuity and royal legitimacy. My emphasis on the socio-political impact of women’s self-representation distinguishes my work from that of scholars such as Mary Fissell and Julie Crawford, who claim that early modern beliefs about the female reproductive body influenced textual depictions of major religious and political events, but give little sustained attention to the role female speech plays in these representations. In contrast, my dissertation reveals that in such texts, patriarchal society relies precisely on the words women speak about their own and other women’s bodies. Ultimately, I argue that female bodily narratives were crucial in shaping early modern culture, and they are equally crucial to our critical understanding of sexual and state politics in the literature of the period.

Uso de suplementos para acasalamento de bezerras Angus aos 14 meses de idade.

RESUMO: O experimento foi conduzido com o objetivo de avaliar o desenvolvimento de novilhas de corte em três sistemas alimentares: exclusivamente em pastagem de azevém (Lolium multiflorum Lam.) ou em pastagem de azevém e recebendo 0,93% do peso corporal de grão de milho inteiro ou grão de aveia branca como suplemento. O delineamento experimental foi o inteiramente ao acaso, com medidas repetidas no tempo. A utilização da pastagem de azevém, com ou sem uso de grão de milho ou aveia como suplemento, possibilita desenvolvimento reprodutivo para que as bezerras sejam acasaladas aos 14 meses de idade. O peso corporal ao início da utilização da pastagem é determinante para que as bezerras atinjam adequado desenvolvimento reprodutivo no início da estação de acasalamento. ABSTRACT: The experiment was carried out to evaluate the development of beef heifers in three feeding systems: exclusively on ryegrass pasture (Lolium multiflorum Lam.) or in ryegrass pasture and receiving 0.93% of body weight of corn whole grain or oats grain as a supplement. The experimental design was completely randomized with repeated measures in time. The utilization of ryegrass pasture, with or without the use of corn or oats grain as a supplement, provides reproductive development of heifers in order for them to be mated at 14 months of age. The body weight at the beginning of pasture utilization is crucial for heifers to reach adequate reproductive development at the beginning of the mating season.

Peptidase activities in the semen from the ductus deferens and uterus of the neotropical rattlesnake Crotalus durissus terrificus

To understand the role of peptidases in seminal physiology of Crotalus durissus terrificus, intra- and inter-seasonal activity levels of acid (APA), basic (APB), puromycin-sensitive (APN-PS) and puromycin-insensitive neutral (APN-PI), cystyl (CAP), dipeptidyl-IV (DPPIV), type-1 pyroglutamyl (PAP-I) and prolyl-imino (PIP) aminopeptidases as well as prolyl endopeptidase (POP) were evaluated in soluble (SF) and/or membrane-bound (MF) fractions of semen collected from the ductus deferens of the male reproductive tract and from the posterior portion of the uterus. Seminal APB, PIP and POP were detected in SF, while other peptidases were detected in SF and MF. Only the convoluted posterior uterus in winter and autumn had semen. Relative to other examined peptidases, in general, APN-PI, APN-PS and APB activities were predominant in the semen from the uterus and throughout the year in the semen from the ductus deferens, suggesting their great relevance in the seminal physiology of C. d. terrificus. The levels of peptidase activities in the ductus deferens semen varied seasonally and were different from those of semen in the uterus, suggesting that their modulatory actions on susceptible peptides are integrated to the male reproductive cycle events and spermatozoa viability of this snake.

Novel guidance cues during neuronal pathfinding in the early scaffold of axon tracts in the rostral brain

A scaffold of axons consisting of a pair of longitudinal tracts and several commissures is established during early development of the vertebrate brain. We report here that NOC-2, a cell surface carbohydrate, is selectively expressed by a subpopulation of growing axons in this scaffold in Xenopus. NOC-2 is present on two glycoproteins, one of which is a novel glycoform of the neural cell adhesion molecule N-CAM. When the function of NOC-2 was perturbed using either soluble carbohydrates or anti-NOC-2 antibodies, axons expressing NOC-2 exhibited aberrant growth at specific points in their pathway. NOC-2 is the first-identified axon guidance molecule essential for development of the axon scaffold in the embryonic vertebrate brain.

A novel two-chain proteinase inhibitor generated by circularization of a multidomain precursor protein

Female reproductive tissues of the ornamental tobacco amass high levels of serine proteinase inhibitors (PIs) for protection against pests and pathogens. These PIs are produced from a precursor protein composed of six repeats each with a protease reactive site. Here we show that proteolytic processing of the precursor generates five single-chain PIs and a remarkable two-chain inhibitor formed by disulfide-bond Linkage of Nand C-terminal peptide fragments. Surprisingly, PI precursors adopt this circular structure regardless of the number of inhibitor domains, suggesting this bracelet-like conformation is characteristic of the widespread potato inhibitor II (Pot II) protein family.

The benefits of polyandry in the free-spawning polychaete Galeolaria caespitosa

In many species, females are thought to benefit from polyandry due to the reduced risks of fertilization by genetically incompatible sperm. However, few studies that have reported such benefits have directly attributed variation in female reproductive success to the interacting effects of males and females at fertilization. In this paper, we determine whether male x female interactions influence fertilization in vitro in the free-spawning, sessile polychaete Galeolaria caespitosa. Furthermore, we determined whether polyandry results in direct fertilization benefits for females by experimentally manipulating the number of males contributing towards staged spawning events. To test for male x female interaction effects we performed an initial experiment that crossed seven males with six females (in all 42 combinations), enabling us to assess fertilization rates for each specific male-female pairing and attribute variation in fertilization success to males, females and their interaction. This initial experiment revealed a strong interaction between males and females at fertilization, confirming that certain male-female combinations were more compatible than others. A second experiment tested the hypothesis that polyandry enhances female reproductive success by exposing each female's eggs to either a single male's sperm (monandry) or the sperm from three males simultaneously (polyandry). We performed this second experiment at two ecologically relevant sperm concentrations. This latter experiment revealed a strong fertilization benefit of polyandry, independent of the effects of sperm concentration (which were also significant). We suggest that these direct fertilization gains arising from polyandry will constitute an important source of selection on females to mate multiply in nature.

Does egg competition occur in marine broadcast-spawners?

When the availability of sperm limits female reproductive success, competition for sperm, may be an important broker of sexual selection. This is because sperm limitation can increase the variance in female reproductive success, resulting in strong selection on females to compete for limited fertilization opportunities. Sperm limitation is probably common in broadcast-spawning marine invertebrates, making these excellent candidates for investigating scramble competition between broods of eggs and its consequences for female reproductive success. Here, we report our findings from a series of experiments that investigate egg competition in the sessile, broadcast-spawning polychaete Galeolaria caespitosa. We initially tested whether the order in which eggs encounter sperm affects their fertilization success at two ecologically relevant current regimes. We used a split-clutch-split-ejaculate technique to compare the fertilization success of eggs from individual females that had either first access (competition-free treatment) or second access (egg competition treatment) to a batch of sperm. We found that fertilization success depended on the order in which eggs accessed sperm; eggs that were assigned to the competition-free treatment exhibited significantly higher fertilization rates than those assigned to the egg competition treatment at both current speeds. In subsequent experiments we found that prior exposure of sperm to eggs significantly reduced both the quantity and quality of sperm available to fertilize a second clutch of eggs, resulting in reductions in fertilization success at high and low sperm concentrations. These findings suggest that female traits that increase the likelihood of sperm-egg interactions (e.g. egg size) will respond to selection imposed by egg competition.

The soybean concentrated extract proliferates the vagina of adult rats

Objective: The aim of this study was to evaluate changes induced on the vagina of ovariectomized rats after treatment with soybean concentrated extract or conjugated equine estrogens and the association of both drugs. Methods: We conducted an experimental study with 50 ovariectomized rats that were randomly divided into five equal groups of 10 animals: GI received vehicle, GII received soybean concentrated extract 46 mg/kg per day, GIII received soybean concentrated extract 120 mg/kg per day, GIV received conjugated equine estrogens 50 mu g/kg per day, and GV received conjugated equine estrogens 50 mu g/kg and soybean concentrated extract 46 mg/kg per day. The substances were administered by gavage during 21 consecutive days. After that, the animals were killed under anesthesia and the vagina was removed for histological and immunohistochemical analysis. Data were initially submitted to analysis of variance. Whenever a significant difference was detected, the study was complemented with the Tukey-Kramer test for multiple comparisons. Results: GII did not show any differences on the vaginal epithelium or collagen compared with GI. GIII presented an increase in vaginal epithelium and collagen amount. GIV had the highest amount of collagen and the signals of vaginal proliferation. GV did not show any additional effect compared with GIV. Conclusions: Our data suggest that a high dose of isoflavone-rich soy extract may have positive effects on the vaginal structures of ovariectomized rats, but this action is less than that of estrogen treatment on vaginal thickness. In addition, soy extract may not block the estrogen effect on vaginal tissue.

Neonatal Handling Reduces Renal Function in Adult Rats

Background/Aims: To evaluate the effects of neonatal handling on hydroelectrolytic balance in adult rats. Methods: The litters were divided into two groups: nonhandled and handled. The procedure consisted of handling the pups for 1 min/day in the first 10 days postnatally. When adults, animals had their body weight verified and were housed in individual metabolic cages. After a 24-hour period, urine samples were collected and the urinary and water intake volumes measured. Blood samples to determine osmolality, aldosterone, corticosterone, angiotensin II, creatinine, urea, sodium and potassium levels were collected. The kidneys were removed for histological assessment. Urinary osmolality, sodium, urea and creatinine were also measured and the creatinine clearance (CC) calculated. Results: No difference between groups was found in the body weight. Handled animals showed a reduction in the total kidney wet weight, water intake, urinary volume, CC, plasma angiotensin II, corticosterone and aldosterone when compared to the nonhandled and an increase in the urinary osmolality and sodium excretion fraction. No differences in serum potassium and no evidence of structural changes were demonstrated by histological analysis. Conclusion: Neonatal handling induced long-lasting effects decreasing renal function without evidence of kidney structural changes. Copyright (c) 2009 S. Karger AG, Basel

Genetic associations between age at first calving and heifer body weight and scrotal circumference in Nelore cattle

P>Age at first calving (AFC) measures the entry of heifers into the beef cattle production system. This trait can be used as a selection criterion for earlier reproductive performance. Using data from Nelore cattle participating in the `Program for Genetic Improvement of the Nelore Breed` (PMGRN-Nelore Brazil), bi-trait analyses were performed using the restricted maximum likelihood method, based on an AFC animal model and the following traits: female body weight adjusted to 365 (BW365) and 450 (BW450) days of age, and male scrotal circumference adjusted to 365 (SC365), 450 (SC450), 550 (SC550) and 730 (SC730) days of age. The heritability estimates for AFC ranged from 0.02 +/- 0.02 to 0.04 +/- 0.02. The estimates of additive direct heritabilities (with standard error) for BW365, BW450, SC365, SC450, SC550 and SC730 were 0.36 +/- 0.07, 0.38 +/- 0.07, 0.48 +/- 0.07, 0.65 +/- 0.07, 0.64 +/- 0.07 and 0.42 +/- 0.07, respectively, and the genetic correlations with AFC were -0.38, -0.33, 0.10, -0.13, -0.13 and 0.06, respectively. In the herds studied, selection for SC365, SC450, SC550 or SC730 should not cause genetic changes in AFC. Selection based on BW365 or BW450 would favor smaller AFC breeding values. However, the low magnitude of direct heritability estimates for AFC in these farms indicates that changes in phenotypical expression depend mostly on non-genetic factors.

Analysis of the Nicotiana tabacum Stigma/Style Transcriptome Reveals Gene Expression Differences between Wet and Dry Stigma Species

The success of plant reproduction depends on pollen-pistil interactions occurring at the stigma/style. These interactions vary depending on the stigma type: wet or dry. Tobacco (Nicotiana tabacum) represents a model of wet stigma, and its stigmas/styles express genes to accomplish the appropriate functions. For a large-scale study of gene expression during tobacco pistil development and preparation for pollination, we generated 11,216 high-quality expressed sequence tags (ESTs) from stigmas/styles and created the TOBEST database. These ESTs were assembled in 6,177 clusters, from which 52.1% are pistil transcripts/genes of unknown function. The 21 clusters with the highest number of ESTs (putative higher expression levels) correspond to genes associated with defense mechanisms or pollen-pistil interactions. The database analysis unraveled tobacco sequences homologous to the Arabidopsis (Arabidopsis thaliana) genes involved in specifying pistil identity or determining normal pistil morphology and function. Additionally, 782 independent clusters were examined by macroarray, revealing 46 stigma/style preferentially expressed genes. Real-time reverse transcription-polymerase chain reaction experiments validated the pistil-preferential expression for nine out of 10 genes tested. A search for these 46 genes in the Arabidopsis pistil data sets demonstrated that only 11 sequences, with putative equivalent molecular functions, are expressed in this dry stigma species. The reverse search for the Arabidopsis pistil genes in the TOBEST exposed a partial overlap between these dry and wet stigma transcriptomes. The TOBEST represents the most extensive survey of gene expression in the stigmas/styles of wet stigma plants, and our results indicate that wet and dry stigmas/styles express common as well as distinct genes in preparation for the pollination process.

Influence of Osteopontin in Bovine Uterine Tube Fluid on Sperm Binding and Fertilization in RCA-1 Lectin-treated Oocytes

This study was conducted to determine the effect of pre-exposure of oocytes to Ricinus communis (RCA-1) lectin and osteopontin (OPN) in uterine tube fluid (UTF) on in vitro sperm-egg binding and fertilization. In vitro-matured bovine oocytes were incubated (39 degrees C, 5% CO(2) in air) for 2 h in the following treatments: (i) 500 mu l of fertilization medium (FM); (ii) 250 mu l of FM with 0.25 ml of non-luteal ampullary uterine tube fluid (NLAUTF); (iii) 250 mu l of FM with 250 mu l of NLAUTF and 4 mu l of RCA-1 lectin; (iv) 250 mu l of FM with 250 mu l of NLAUTF, a rabbit polyclonal antibody (1:200) against purified bovine milk OPN, and RCA-1 lectin; (v) 500 mu l of FM and RCA-1 lectin. Following incubation, oocytes were washed, placed in FM with 2 mu g heparin, and incubated with 1 x 10(5) frozen-thawed spermatozoa per 10 oocytes. Oocytes used to assess sperm binding were stained with Hoescht 33342, and the number of sperm bound per zona pellucida counted. The remaining oocytes were fixed in acid alcohol, stained with 1% acetate-orcein and observed to determine the presence of pronuclei. More sperm bound to the zona pellucida (mean +/- SEM) when oocytes were incubated in treatment 3 (59.0 +/- 5.5) than in treatments 2 (46.4 +/- 5.6), 4 (18.1 +/- 5.4), 5 (33.4 +/- 5.6) or 1 (32.5 +/- 5.6). More oocytes were fertilized when incubated in treatment 3 (91% +/- 3.0) than in 2 (84% +/- 3.0), 4 (40% +/- 3.0), 5 (77% +/- 3.0) or 1 (76% +/- 3.0). As in previous studies, this study suggests that RCA-1 lectin enhances binding of UTF-derived OPN to bovine oocytes, resulting in increased sperm-egg binding and fertilization in vitro and a possible role in fertilization.