ESTRESSE, DEPRESSÃO, SATISFAÇÃO SEXUAL, VIVÊNCIAS EMOCIONAIS E QUALIDADE DE VIDA EM PACIENTES DE AMBULATÓRIOS DE REPRODUÇÃO HUMANA

Este estudo teve como objetivos avaliar a Qualidade de Vida (QV), sintomas depressivos, estresse, satisfação sexual e vivências emocionais em pacientes que apresentam infertilidade. Participaram deste estudo 500 pessoas (mulheres e homens), com idade igual ou maior a 18 anos, pacientes de um ambulatório de reprodução humana localizado no Grande ABC Paulista. O estudo utilizou instrumentos de avaliação autoaplicáveis: WHOQOL-Bref, BDI-II, ISQ, ICQ, ISS e questionário sócio-demográfico. A caracterização da amostra estudada demostrou alto nível escolar (62,4pc), boa condição econômica (80,2pc, idade superior a 30 anos (82,8pc) e a maioria apresenta infertilidade primária (73,2pc). Quanto à QV, níveis de depressão e estresse os homens apresentam melhores resultados quando comparados às mulheres. No que ser refere aos níveis de satisfação sexual 56,6% dos participantes descrevem ter satisfação sexual, mas 45,4% afirmam ter insatisfação sexual e não houve diferença entre os gêneros. As análises dos dados mostraram que é maior o número de mulheres do que o de homens que procuram atendimento para infertilidade (64,8pc). Pôde-se comprovar que além das dificuldades provenientes da infertilidade, o tratamento, também, impacta sobre a vida dessas pessoas. Foram diversos participantes que descreveram significativas mudanças (42,8pc), dos quais apontaram em maioria absoluta (92,6pc) uma piora no estado de humor devido ao tratamento. Como também é alto o índice de pessoas que descrevem sintomas de depressão (38,8pc) e estresse (31,4pc) acima da média da população geral. Ao considerar as especificidades da qualidade de vida averiguou-se um maior impacto no domínio psicológico dos pacientes (15,20 pontos) comparado ao físico (15,68 pontos). Estes resultados ressaltam a importância do atendimento psicológico para as pessoas em programas de reprodução humana.

Mouse embryo culture induces changes in postnatal phenotype including raised systolic blood pressure

A key factor in the use of assisted reproductive technologies (ART) for diverse species is the safety of procedures for long-term health. By using a mouse model, we have investigated the effect of in vitro culture and embryo transfer (ET) of superovulated embryos on postnatal growth and physiological activity compared with that of embryos developing in vivo. Embryo culture from two-cell to blastocyst stages in T6 medium either with or without a protein source reduced blastocyst trophectoderm and inner cell mass cell number compared with that of embryos developing in vivo. Embryo culture and ET had minimal effects on postnatal growth when compared with in vivo development with an equivalent litter size. However, embryo culture, and to a lesser extent ET, led to an enhanced systolic blood pressure at 21 weeks compared with in vivo development independent of litter size, maternal origin, or body weight. Moreover, activity of enzymatic regulators of cardiovascular and metabolic physiology, namely, serum angiotensin-converting enzyme and the gluconeogenesis controller, hepatic phosphoeno/pyruvate carboxykinase, were significantly elevated in response to embryo culture and/or ET in female offspring at 27 weeks, independent of maternal factors and postnatal growth. These animal data indicate that postnatal physiological criteria important in cardiovascular and metabolic health may be more sensitive to routine ART procedures than growth. © 2007 by The National Academy of Sciences of the USA.

Una reflexión prospectiva sobre técnicas sostenibles para controlar la reproducción en mamíferos domésticos

El control de la reproducción de los animales de granja fue una de las herramientas esenciales en la domesticación. Esto es aún de gran interés para la mejora genética, el ajuste de la producción a la disponibilidad de alimentos y del mercado y finalmente, para la reducción de los períodos improductivos. La detección del comportamiento del estro, la sincronización de las concepciones y el incremento de la potencialidad de difusión de los padres de alta genética son tres objetivos comunes a la producción de todas las especies. Los variados sistemas reproductivos entre los diversos sistemas de producción difieren a causa de las propiedades intrínsecas de las especies y de los diferentes grados de intensificación de estos sistemas. Tres tendencias claras son el continuo incremento de la productividad por medio de las mejoras de la eficiencia reproductiva, el desarrollo de técnicas nuevas y sostenibles sin el uso de hormonas y el continuo desarrollo de la inseminación artificial y de las biotécnicas reproductivas. Las futuras áreas de inversión en investigación podrían ser: las bases fisiológicas y etológicas de las interrelaciones socio-sexuales entre animales, el control genético de parámetros reproductivos, el incremento en la eficiencia de biotecnologías clásicas y nuevas y la ingeniería de técnicas reproductivas nuevas e innovativas para ser utilizadas a nivel de granja. Estas técnicas reproductivas deberían ser desarrolladas respetando los tres pilares de la sostenibilidad: el ambiente, la economía y la sociedad. Por ello deberían estar incluidas dentro de los sistemas de producción en los cuales se supone que serán aplicadas y donde deberían ser evaluadas en su sostenibilidad.

Development and characterisation of solution processed vanadium oxide and transparent metal oxide thin films

Metal oxide thin films are important for modern electronic devices ranging from thin film transistors to photovoltaics and functional optical coatings. Solution processed techniques allow for thin films to be rapidly deposited over a range of surfaces without the extensive processing of comparative vapour or physical deposition methods. The production of thin films of vanadium oxide prepared through dip-coating was developed enabling a greater understanding of the thin film formation. Mechanisms of depositing improved large area uniform coverage on a number of technologically relevant substrates were examined. The fundamental mechanism for polymer-assisted deposition in improving thin film surface smoothness and long range order has been delivered. Different methods were employed for adapting the alkoxide based dip-coating technique to produce a variety of amorphous and crystalline vanadium oxide based thin films. Using a wide range of material, spectroscopic and optical measurement techniques the morphology, structure and optoelectronic properties of the thin films were studied. The formation of pinholes on the surface of the thin films, due to dewetting and spinodal effects, was inhibited using the polymer assisted deposition technique. Uniform thin films with sub 50 nm thicknesses were deposited on a variety of substrates controlled through alterations to the solvent-alkoxide dilution ratios and employing polymer assisted deposition techniques. The effects of polymer assisted deposition altered the crystallized VO thin films from a granular surface structure to a polycrystalline structure composed of high density small in-plane grains. The formation of transparent VO based thin film through Si and Na substrate mediated diffusion highlighted new methods for material formation and doping.

Le nuove forme di genitorialità tra vincoli biologici, verita legale e legami affettivi

Le recenti novelle e l’evoluzione delle tecniche riproduttive scientifiche hanno promosso un progressivo processo di frammentazione della figura genitoriale, che sino ad allora era considerata unitaria ed intimamente connessa all’incontro tra uomo e donna. A tale concezione unitaria del legame genitoriale corrispondeva la considerazione che la coincidenza tra vincolo genetico e verità legale fosse foriera del soddisfacimento del superiore interesse del minore. La genitorialità può ad oggi essere suddivisa tra genetica, qualora tra il genitore condivida con il figlio il medesimo patrimonio genetico; biologica, nell’accezione di gestazionale e dunque caratterizzante il rapporto tra madre e figlio; e sociale, qualora l’adulto, seppur in mancanza di un titolo legale e altrettanto di un legame di sangue con il figlio, adempie ai doveri di cura e di mantenimento propri dell’esercente la responsabilità genitoriale. A fronte della pluralità dei soggetti che latu senso possono essere considerati genitoriali in forza di una pluralità e eterogeneità di legami, l’interprete deve interrogarsi circa la effettività e la cogenza delle tradizioni definizioni di madre e di padre e soprattutto la resilienza dei criteri attributivi dello stato di filiazione.

Computer-assisted cup placement techniques in total hip arthroplasty improve accuracy of placement.

Malposition of the acetabular component during hip arthroplasty increases the occurrence of impingement, reduces range of motion, and increases the risk of dislocation and long-term wear. To prevent malpositioned hip implants, an increasing number of computer-assisted orthopaedic systems have been described, but their accuracy is not well established. The purpose of this study was to determine the reproducibility and accuracy of conventional versus computer-assisted techniques for positioning the acetabular component in total hip arthroplasty. Using a lateral approach, 150 cups were placed by 10 surgeons in 10 identical plastic pelvis models (freehand, with a mechanical guide, using computer assistance). Conditions for cup implantations were made to mimic the operating room situation. Preoperative planning was done from a computed tomography scan. The accuracy of cup abduction and anteversion was assessed with an electromagnetic system. Freehand placement revealed a mean accuracy of cup anteversion and abduction of 10 degrees and 3.5 degrees, respectively (maximum error, 35 degrees). With the cup positioner, these angles measured 8 degrees and 4 degrees (maximum error, 29.8 degrees), respectively, and using computer assistance, 1.5 degrees and 2.5 degrees degrees (maximum error, 8 degrees), respectively. Computer-assisted cup placement was an accurate and reproducible technique for total hip arthroplasty. It was more accurate than traditional methods of cup positioning.

Imaging techniques applied for detection of secretion, transgene delivery and G proteincoupled receptors in reproductive and endocrine cells in vivo and in vitro

Post-testicular sperm maturation occurs in the epididymis. The ion concentration and proteins secreted into the epididymal lumen, together with testicular factors, are believed to be responsible for the maturation of spermatozoa. Disruption of the maturation of spermatozoa in the epididymis provides a promising strategy for generating a male contraceptive. However, little is known about the proteins involved. For drug development, it is also essential to have tools to study the function of these proteins in vitro. One approach for screening novel targets is to study the secretory products of the epididymis or the G protein-coupled receptors (GPCRs) that are involved in the maturation process of the spermatozoa. The modified Ca2+ imaging technique to monitor release from PC12 pheochromocytoma cells can also be applied to monitor secretory products involved in the maturational processes of spermatozoa. PC12 pheochromocytoma cells were chosen for evaluation of this technique as they release catecholamines from their cell body, thus behaving like endocrine secretory cells. The results of the study demonstrate that depolarisation of nerve growth factor -differentiated PC12 cells releases factors which activate nearby randomly distributed HEL erythroleukemia cells. Thus, during the release process, the ligands reach concentrations high enough to activate receptors even in cells some distance from the release site. This suggests that communication between randomly dispersed cells is possible even if the actual quantities of transmitter released are extremely small. The development of a novel method to analyse GPCR-dependent Ca2+ signalling in living slices of mouse caput epididymis is an additional tool for screening for drug targets. By this technique it was possible to analyse functional GPCRs in the epithelial cells of the ductus epididymis. The results revealed that, both P2X- and P2Y-type purinergic receptors are responsible for the rapid and transient Ca2+ signal detected in the epithelial cells of caput epididymides. Immunohistochemical and reverse transcriptase-polymerase chain reaction (RTPCR) analyses showed the expression of at least P2X1, P2X2, P2X4 and P2X7, and P2Y1 and P2Y2 receptors in the epididymis. Searching for epididymis-specific promoters for transgene delivery into the epididymis is of key importance for the development of specific models for drug development. We used EGFP as the reporter gene to identify proper promoters to deliver transgenes into the epithelial cells of the mouse epididymis in vivo. Our results revealed that the 5.0 kb murine Glutathione peroxidase 5 (GPX5) promoter can be used to target transgene expression into the epididymis while the 3.8 kb Cysteine-rich secretory protein-1 (CRISP-1) promoter can be used to target transgene expression into the testis. Although the visualisation of EGFP in living cells in culture usually poses few problems, the detection of EGFP in tissue sections can be more difficult because soluble EGFP molecules can be lost if the cell membrane is damaged by freezing, sectioning, or permeabilisation. Furthermore, the fluorescence of EGFP is dependent on its conformation. Therefore, fixation protocols that immobilise EGFP may also destroy its usefulness as a fluorescent reporter. We therefore developed a novel tissue preparation and preservation techniques for EGFP. In addition, fluorescence spectrophotometry with epididymal epithelial cells in suspension revealed the expression of functional purinergic, adrenergic, cholinergic and bradykinin receptors in these cell lines (mE-Cap27 and mE-Cap28). In conclusion, we developed new tools for studying the role of the epididymis in sperm maturation. We developed a new technique to analyse GPCR dependent Ca2+ signalling in living slices of mouse caput epididymis. In addition, we improved the method of detecting reporter gene expression. Furthermore, we characterised two epididymis-specific gene promoters, analysed the expression of GPCRs in epididymal epithelial cells and developed a novel technique for measurement of secretion from cells.

Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Characterization of Primary Amine End-Functionalized Polystyrene and Poly(methyl methacrylate) Synthesized by Living Anionic Polymerization Techniques

The reaction of living anionic polymers with 2,2,5,5-tetramethyl-1-(3-bromopropyl)-1-aza-2,5- disilacyclopentane (1) was investigated using coupled thin layer chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Structures of byproducts as well as the major product were determined. The anionic initiator having a protected primary amine functional group, 2,2,5,5-tetramethyl- 1-(3-lithiopropyl)-1-aza-2,5-disilacyclopentane (2), was synthesized using all-glass high-vacuum techniques, which allows the long-term stability of this initiator to be maintained. The use of 2 in the preparation of well-defined aliphatic primary amine R-end-functionalized polystyrene and poly(methyl methacrylate) was investigated. Primary amino R-end-functionalized poly(methyl methacrylate) can be obtained near-quantitatively by reacting 2 with 1,1-diphenylethylene in tetrahydrofuran at room temperature prior to polymerizing methyl methacrylate at -78 °C. When 2 is used to initiate styrene at room temperature in benzene, an additive such as N,N,N',N'- tetramethylethylenediamine is necessary to activate the polymerization. However, although the resulting polymers have narrow molecular weight distributions and well-controlled molecular weights, our mass spectra data suggest that the yield of primary amine α-end-functionalized polystyrene from these syntheses is very low. The majority of the products are methyl α-end-functionalized polystyrene.

Interventional Laser-assisted Extraction of an Infected Trapped Ventriculoatrial Shunt Based on Techniques for Pacemaker Lead Explantation.

We present a case of successful interventional laser-assisted extraction of an endovascularly trapped long-term implanted ventriculoatrial shunt in a patient with shunt-associated septicemia. This approach is based on modified techniques for explantation of pacemaker leads and might be considered an additional option for extraction of otherwise nonremovable trapped endovascular catheters in experienced centers.

Computer-assisted virtual autopsy using surgical navigation techniques

UNLABELLED OBJECTIVE; Virtual autopsy methods, such as postmortem CT and MRI, are increasingly being used in forensic medicine. Forensic investigators with little to no training in diagnostic radiology and medical laypeople such as state's attorneys often find it difficult to understand the anatomic orientation of axial postmortem CT images. We present a computer-assisted system that permits postmortem CT datasets to be quickly and intuitively resliced in real time at the body to narrow the gap between radiologic imaging and autopsy. CONCLUSION Our system is a potentially valuable tool for planning autopsies, showing findings to medical laypeople, and teaching CT anatomy, thus further closing the gap between radiology and forensic pathology.

Use of ovary culture techniques in reproductive toxicology

Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved. Acknowledgements The author's studies in this field are supported by MRC grants G1002118 (NS and RAA) and G110357 (RAA), MR/L010011/1 (PAF), the European Community's Seventh Framework Programme (FP7/2007–2013) under grant agreement no. 212885 (PAF) and the Wellcome Trust (080388 to PAF). AS was funded by a BBSRC CASE Studentship co-funded by AstraZeneca.

Endometrial claudin-4 and leukemia inhibitory factor are associated with assisted reproduction outcome

Background: Claudin-4 (CLDN4) is one of several proteins that act as molecular mediators of embryo implantation. Recently, we examined immunolabeling of leukemia inhibitory factor (LIF) in the endometrial tissue of 52 IVF patients, and found that LIF staining intensity was strongly correlated with successful pregnancy initiation. In the same set of patients, we have now examined endometrial CLDN4 expression, to see how expression intensity may vary with LIF. We examined CLDN4 in the luteal phase of the menstrual cycle, immediately preceding IVF treatment. Our aim was to compare expression of LIF and CLDN4 in the luteal phase, and document these patterns as putative biomarkers for pregnancy. Methods: Endometrial tissue was collected from women undergoing IVF. Endometrial biopsies were obtained during the luteal phase preceding IVF, and were then used for tissue microarray (TMA) immunolabeling of CLDN4. Previously published LIF expression data were then combined with CLDN4 expression data, to determine CLDN4/LIF expression patterns. Associations between successful pregnancy after IVF and combined CLDN4/LIF expression patterns were evaluated. Results: Four patterns of immunolabeling were observed in the endometrial samples: 16% showed weak CLDN4 and strong LIF (CLDN4(-)/LIF(+)); 20% showed strong CLDN4 and strong LIF (LIF(+)/CLDN4(+)); 28% showed strong CLDN4 and weak LIF (CLDN4(+)/LIF(-)); and 36% showed weak CLDN4 and weak LIF (CLDN4(-)/LIF(-)). Successful implantation after IVF was associated with CLDN4(-)/LIF(+)(p = 0.003). Patients showing this endometrial CLDN4(-)/LIF(+) immunolabeling were also 6 times more likely to achieve pregnancy than patients with endometrial CLDN4(+)/LIF(-) immunolabeling (p = 0.007). Conclusion: The combined immunolabeling expression of CLDN4(-)/LIF(+) in endometrial tissue is a potential biomarker for predicting successful pregnancy in IVF candidates.

Reproductive Activity and Survivorship of Liriomyza sativae (Diptera: Agromyzidae) at Different Temperatures and Relative Humidity Levels

We studied a population of Liriomyza sativae Blanchard (Diptera: Agromyzidae) identified by morphological and molecular techniques from the semiarid region of the Brazilian northeast. The influence of temperature and relative humidity on the survival and reproductive parameters of L. sativae in cowpeas (Vigna unguiculata L. Walp.) (Fabales: Fabaceae) was evaluated. We used temperatures of 18, 20, 22, 25, 28, 30, and 32 +/- 1 degrees C (50 +/- 10% RH) and relative humidity values of 30,50,70, and 90 +/- 10% (25 +/- 1 degrees C) under a 14 L:10 D photoperiod. Adult longevity decreased as temperature and relative humidity increased and was greater, in general, for females. The preoviposition and oviposition periods also decreased as temperature increased, whereas relative humidity only caused reductions in the oviposition period at higher levels. Fecundity was similar in the range from 18 to 30 degrees C but decreased at 32 degrees C with respect to relative humidity; the best performances of L. sativae occurred at lower levels. The pattern of oviposition rate changed with temperature and relative humidity. Regardless of temperature and relative humidity, L, sativae laid between 75 and 92% of its eggs on the adaxial surface of the cowpea leaves. This information will be highly useful to design a leafminer production system aimed at the multiplication of natural enemies, as well as for pest management in the field.

Adhesion of microbes using 3-aminopropyl triethoxy silane and specimen stabilisation techniques for analytical transmission electron microscopy

A variety of adhesive support-films were tested for their ability to adhere various biological specimens for transmission electron microscopy. Support films primed with 3-amino-propyl triethoxy silane (APTES), poly-L-lysine, carbon and ultraviolet-B (UV-B)-irradiated carbon were tested for their ability to adhere a variety of biological specimens including axenic cultures of Bacillus subtilis and Escherichia coli and wild-type magnetotactic bacteria. The effects of UV-B irradiation on the support film in the presence of air and electrostatic charge on primer deposition were tested and the stability of adhered specimens on various surfaces was also compared. APTES-primed UV-B-irradiated Pioloform(TM) was consistently the best adhesive, especially for large cells, and when adhered specimens were UV-B irradiated they became remarkably stable under an electron beam. This assisted the acquisition of in situ phase-contrast lattice images from a variety of biominerals in magnetotactic bacteria, in particular metastable greigite magnetosomes. Washing tests indicated that specimens adhering to APTES-primed UV-B-irradiated Pioloform(TM) were covalently coupled. The electron beam stability was hypothesised to be the result of mechanical strengthening of the specimen and support film and the reduced electrical resistance in the specimen and support film due to their polymerization and covalent coupling.