A Novel Treatment for Convex Lateral Crura: The Inverted Spanning Suture.

Rhinoplasty is one of the most complex and challenging procedures in plastic surgery, even for experienced surgeons. Among the steps leading to an aesthetic and functional nose, there is the nasal tip improvement. Today's approach to nasal tip is the product of different techniques shifting through time, mainly from a resection tendency, to preservation and suture use to reshape cartilages. Addressing the lateral crura is vital to an aesthetic nasal tip and it is frequently obtained by adequate suture techniques. The alar-spanning suture described by Perkins is one of such. Regardless of its importance, it was not able to improve convex crura in some cases. The inverted alar-spanning suture (ISS) is an adaptation designed to treat those cases with the suture alone. ISS is a novel technique that can lead to better results treating the convex lateral crura by distributing the force vector in a more effective way. New techniques in rhinoplasty have multiplied, bringing this procedure to a new level and keeping up with the updated notion of restoration instead of excision the ISS is a new, precise, approach to an old problem.

In vitro and in vivo embryo production efficiency in Flemish and Holstein donor females.

The aim of this study was to compare embryo production efficiency in Flemish and Holstein donor females using ovum pick-up and in vitro fertilization (OPU-IVF) or in vivo production (superovulation; SOV) procedures. The study was conducted using a split-plot design, with eight Flemish and eight Holstein non-lactating cycling females. Females were subjected to ten weekly OPU/IVF sessions and/or two SOV/embryo collections sessions at a 63-day interval, for a total of 160 OPU-IVF and 32 SOV sessions. Mean numbers of follicles and corpora lutea, and cumulus-oocyte complex (COC) recovery rates were similar between breeds after the OPU and SOV sessions. However, Flemish donors yielded better quality grade II COCs (301, 41.9%) than Holstein females (609, and 202, 33.1%). Also, cleavage and blastocyst rates, and the total number and the mean number of viable embryos obtained after OPU-IVF were higher in Flemish (49.6% and 11.8%, and 63 and 11.8 per donor, respectively) than in Holstein (32.8% and 7.2%, and 34 and 7.2 per donor, respectively) females. Flemish females were also more efficient in yielding viable embryos after SOV (111, 7.3 per donor) than Holstein (48, 3.3 per donor) females. Overall, Flemish donor females had better responses to OPU-IVF or SOV procedures than Holstein counterparts. Irrespective of the breeds, SOV procedures were more efficient than OPU-IVF in yielding more viable embryos, under the conditions of this study. Both reproductive procedures were useful tools for the genetic conservation of the Flemish cattle breed in Southern Brazil.

ß-Mercaptoethanol in culture medium improves cryotolerance of in vitro-produced bovine embryos.

The objective of this study was to investigate the effects of adding ß-mercaptoethanol (ßME) to culture medium of bovine in vitro-produced (IVP) embryos prior to or after vitrification on embryo development and cryotolerance. In Experiment I, Day-7 IVP blastocysts were vitrified and, after warming, cultured in medium containing 0, 50 or 100 µM ßME for 72 h. Embryos cultured in 100 µM ßME attained higher hatching rates (66.7%) than those culture in 0 (47.7%) and 50 (52.4%) µM ßME. In Experiment II, IVP embryos were in vitro-cultured (IVC) to the blastocyst stage in 0 (control) or 100 µM ßME, followed by vitrification. After warming, embryos were cultured for 72 h (post-warming culture, PWC) in 0 (control) or 100 µM ßME, in a 2 × 2 factorial design: (i) CTRL-CTRL, control IVC and control PWC; (ii) CTRL-ßME, control IVC and ßME-supplemented PWC; (iii) ßME-CTRL, ßME-supplemented IVC and control PWC; or (iv) ßME-ßME, ßME-supplemented IVC and ßME-supplemented PWC. ßME during IVC reduced embryo development (28.0% vs. 43.8%) but, following vitrification, higher re-expansion rates were seen in ßME-CTRL (84.0%) and ßME-ßME (87.5%) than in CTRL-CTRL (71.0%) and CTRL-ßME (73.1%). Hatching rates were higher in CTRL-ßME (58.1%) and ßME-ßME (63.8%) than in CTRL-CTRL (36.6%) and ßME-CTRL (42.0%). Total cell number in hatched blastocysts was higher in ßME-ßME (181.2 ± 7.4 cells) than CTRL-CTRL (139.0 ± 9.9 cells). Adding ßME to the IVC medium reduced development but increased cryotolerance, whereas adding ßME to the PWC medium improved embryo survival, hatching rates, and total cell numbers.

Components of the insulin-like growth factor system in in vivo - and in vitro-derived fetuses of cattle, and the association with growth and development.

This experiment was designed to study mechanisms affecting growth of in vivo-derived (IVD) and in vitro-produced (IVP) fetuses of cattle. Day-7 IVD or IVP cattle blastocysts were transferred to recipients, with pregnant females being slaughtered on Days 90 or 180 of gestation or allowed to undergo parturition. Uteri and contents were dissected and physically measured, and maternal and fetal plasma and amniotic and allantoic fluids were collected for IGF-1 and IGF-2 determinations, and IGFBP profile characterization. Transcripts for IGF-1 and IGF-2 mRNA in placental and fetal tissues, and IGF-1r and IGF-2r in placentomes were determined. There was a greater fetal weight in the IVP group, which was associated with greater IGF-1 and IGF-2 concentrations in maternal circulation, and changes in IGFBP profiles within fetal fluids. Day-90 IVP-derived fetuses were longer, had greater organ weights, larger placentomes, less placentome IGF-2r mRNA transcript, and greater maternal IGF-1 and IGF-2 concentrations than controls. On Day 180 and at parturition tissues from IVP-derived fetuses/calves were from larger uteri, with larger placentomes/fetal membranes, fetuses/calves weighed more, had greater fetal hepatic IGF-2 mRNA transcript, had less fetal plasma IGF-1 and greater allantoic IGF-2 concentrations, greater and lesser IGFBP activities in the allantoic and amniotic fluids, respectively, and greater glucose and fructose accumulation in fetal fluids. Components of the IGF system were differentially regulated not only according to the gestation period (Days 90 or 180) and fluid type (maternal or fetal plasma, amniotic or allantoic fluids), but also based on conceptus origin (IVP or IVD) in cattle.

The Horn Technique for Nasal Tip Support in Rhinoplasty.

Introduction The predictability of nasal tip projection and rotation after aesthetic surgery is a challenge. Tongue-in-groove (TIG) is an effective technique to control tip projection and rotation, but there may be a small loss of projection and rotation of the tip lobe due to lack of support between the anterior septal angle and the domus, since this region is sustained by medial crusts suture-linked and interdomus sutures. Objective To describe a new surgery technique in an attempt to correct the lack of support for the nasal tip after lowering the nasal dorsum. Methods The horn technique consists in preserving a square of cartilage during the removal of the nasal dorsum and septum excess in patients with long and projected nose. This piece will give greater support to the TIG technique and greater predictability of the rotation and projection of the nasal tip. Results Between 2016 and 2018, 50 patients with long and projected noses were submitted to the "horn technique" surgery. They were submitted to the TIG technique associated to the horn technique. A retrospective review of the preoperative and postoperative photographs (3 months to 1 year) of these patients treated with the horn technique were analyzed and showed better support of the nasal tip. Conclusion The horn technique provides greater support to the projection and rotation of rhinoplasties in patients with long and projected nose.

Promoter-specific expression of the imprinted IGF2 gene in bovine oocytes and pre-implantation embryos.

The bovine IGF2 locus is a genomic region with alternative transcripts controlled by five promoters (P0, P1, P2, P3 and P4). As transcriptional regulation can affect messenger RNA (mRNA) stability and translation, and thus, subsequent biological effects, this study evaluated the bovine IGF2 promoter-specific expression patterns in oocytes and pre-implantation embryos produced in vitro by our standard IVP procedures. Immature and matured oocytes, and pre-implantation embryos at the 1-, 2-, 4-, 8- and 16-cell, and at early morula, compact morula, blastocyst and expanded blastocyst stages were collected in three pools of five structures per stage, in four replicates. Total RNA was extracted and subjected to RT-qPCR, using four sets of IGF2 promoter-specific primers covering transcripts driven by promoters P0/P1, P2, P3 and P4, with fragments sequenced for confirmation. Expression of P2- and P4-derived transcripts showed an initial peak between immature (P4) or matured (P2/P4) oocytes and 2-cell embryos, gradually falling until embryo genome activation (EGA), rising again at compaction and cavitation. P0/P1-derived transcripts were identified after EGA, during compaction, whereas P3 activity was not detected at any stage. Our findings suggest that P0/P1 and P2 likely have secondary roles during early stages, whereas P3 may be more relevant later in development. P4 seems to be the main pathway for bovine IGF2 expression during oocyte maturation and embryo development and, therefore, the main target to influence IVP in modulation of embryo growth and in studies in developmental biology.

Assessment of binder of sperm protein 1 (BSP1) and heparin effects on in vitro capacitation and fertilization of bovine ejaculated and epididymal sperm.

The present study evaluated the effect of binder of sperm protein 1 (BSP1) and/or heparin on in vitro bovine capacitation and fertilization rates using epididymal and ejaculated bovine sperm. Frozen-thawed sperm were selected and used in the following treatments. Control group: Fert-TALP medium without heparin; heparin (HEP) group: Fert-TALP with heparin (10 UI/ml); BSP1 group: Fert-TALP medium with BSP1 (10 µg/ml for ejaculated sperm; 40 µg/ml for epididymal sperm); HEP + BSP1 group: Fert-TALP medium with heparin (5 UI/ml) and BSP1 (5 µg/ml for ejaculated sperm; 20 µg/ml for epididymal sperm) and determined in vitro capacitation rates in different interval times (0, 15, 30 and 60 min) using the chlortetracycline fluorescence (CTC) method. Also, we evaluated the development rates of oocytes fertilized with ejaculated or epididymal sperm into the same treatments. Capacitation was greater and faster when ejaculated sperm were treated for 60 min with heparin compared with other treatments. However, developmental rates were similar in all treatments. For epididymal sperm, the treatments with BSP1 presented higher capacitation and fertilization rates compared with heparin (P < 0.05). The effects of heparin + BSP1 on capacitation and developmental rates did not cause any increase in capacitation or blastocyst rates compared with other groups for ejaculated or epididymal sperm. In conclusion, this study confirmed that either BSP1 and heparin can be used as capacitator agents for bovine ejaculated sperm during IVF. However, BSP1 seems to be more efficient compared with heparin for epididymal sperm. Furthermore, BSP1 and heparin have no synergic effects on sperm capacitation.

Ginástica rítmica: experiência baseada nos esportes unificados, da Special Olympics / Rhythmic gymnastics: experience based on unified sports, Special Olympics / Gimnasia rítmica: experiencia basada en los deportes unificados de las Olimpiadas Especial

O programa de Esporte Unificado, da Special Olympics, possibilita práticas esportivas para pessoas com e sem deficiência intelectual conjuntamente, visando a promoção da inclusão social. Fundamentado neste programa, foi realizada uma pesquisa-ação objetivando analisar a relação entre atletas com e sem deficiência intelectual em aulas de ginástica rítmica fundamentadas no Esporte Unificado. Para isso, 18 meninas, 12 com deficiência intelectual e 6 sem essa condição, realizaram aulas de ginástica rítmica, conjuntamente, por 12 semanas, com 2 sessões semanais de 2 horas cada, em uma quadra de esportes de Guaratinguetá, São Paulo, Brasil. Ao final do programa, foi empregada uma entrevista semiestruturada com as meninas sem deficiência, apreciando os dados por análise de conteúdo. Os resultados apontaram que a interação entre atletas com e sem deficiência promoveu melhoras na performance das atletas e possibilitou a construção de conhecimento sobre as diversidades, alcançando atitudes de aceitação e respeito pelas diferenças.

The Special Olympics Unified Sport program enables sports practices for people with and without intellectual disabilities to work together to promote social inclusion. Based on this program, an action research was carried out aiming to verify possible implications achieved through the implementation of a rhythmic gymnastics project based on the Unified Sport, from the perspective of non-disabled participants. To that end, 18 girls, 12 with intellectual disabilities and 6 without this condition, performed rhythmic gymnastics classes together for 12 weeks, with 2 weekly sessions of 2 hours each, in a sports court in Guaratinguetá, São Paulo, Brazil. At the end of the program, a semi-structured interview with non-disabled girls was used, assessing the data by content analysis. The results showed improvements in the performance of athletes, stimulation of the interactions between athletes with and without disabilities and the construction of knowledge about the diversities, reaching attitudes of acceptance and respect for differences.

El programa de deportes unificados de Olimpiadas Especiales permite prácticas deportivas para personas con y sin discapacidad intelectual conjuntamente, con vistas a la promoción de la inclusión social. Fundamentado en este programa, se realizó una investigación-acción objetivando verificar posibles implicaciones alcanzadas por medio de la implantación de un proyecto de gimnasia rítmica fundamentado en el Deporte Unificado, desde la perspectiva de los partícipes sin discapacidad. Para ello, 18 niñas, 12 con discapacidad intelectual y 6 sin esa condición, realizaron clases de gimnasia rítmica, conjuntamente, por 12 semanas, con 2 sesiones semanales de 2 horas cada una, en una cuadra de deportes de Guaratinguetá, São Paulo, Brasil. Al final del programa, se empleó una entrevista semiestructurada con las niñas sin discapacidad, apreciando los datos por análisis de contenido. Los resultados apuntaron mejoras en el desempeño de las atletas, estímulo a las interacciones entre atletas con y sin discapacidad ya la construcción de conocimiento sobre las diversidades, alcanzando actitudes de aceptación y respeto por las diferencias.

In vitro development of IVF-derived bovine embryos following cytoplasmic microinjection for the episomal expression of the IGF2 gene.

Important genomic imprinting changes usually occur following the in vitro production (IVP) of bovine embryos, especially in the imprinting pattern of components of the IGF system. This study aimed to evaluate the effects of a transient episomal overexpression of the IGF2 gene in bovine IVP embryos following embryo cytoplasmic microinjection (CMI) at the 1-cell stage on embryo survival, early and late developmental kinetics and morphological quality up to Day 7 of development. Selected cumulus-oocyte complexes (COCs) were matured and fertilized in vitro and subsequently segregated into six experimental groups: non-CMI control group and five CMI groups at increasing doses (0, 10, 20, 40 and 80 ng/µl) of a GFP vector built for the episomal expression of bovine IGF2. Zygote CMI was effective in delivering the expression vector into the ooplasm, irrespective of the groups, with 58% of positive GFP fluorescence in Day 7 blastocysts. Considering developmental rates and late embryo kinetics, the 10-ng/µl CMI vector dose promoted a lower blastocyst rate (10.4%), but for blastocysts at more advanced stages of development (93.0% blastocysts and expanded blastocysts), and higher number of cells (116.0 ± 3.0) than non-CMI controls (23.3%, 75.0% and 75.0 ± 6.8 were obtained, respectively). In conclusion, CMI at the 1-cell stage did not compromise subsequent in vitro development of surviving embryos, with the 10-ng/µl group demonstrating a possible growth-promoting effect of the IGF2 gene on embryo development, from the 1-cell to the blastocyst stage.

Effects of fusion-activation interval and embryo aggregation on in vitro and in vivo development of bovine cloned embryos.

Nuclear reprogramming in somatic cell cloning is one of the key factors for proper development, with variations in the protocol appearing to improve cloning efficiency. This study aimed to determine the effects of two fusion-activation intervals and the aggregation of bovine cloned embryos on subsequent in vitro and in vivo development. Zygotes produced by handmade cloning were exposed to two fusion-activation intervals (2 h or 4 h), and then cultured in microwells either individually (1 × 100%) or after aggregation of two structures (2 × 100%). Zona-intact oocytes and zona-free oocytes and hemi-oocytes were used as parthenote controls under the same fusion-activation intervals. Day-7 cloned blastocysts were transferred to synchronous recipients. Cleavage (Day 2), blastocyst (Day 7) and pregnancy (Day 30) rates were compared by the χ2 test (P < .05). Extending fusion-activation interval from 2 to 4 h reduced cleavage (91.0 vs. 74.4%) but not blastocyst (34.8 vs. 42.0%) rates. On a microwell basis, cloned embryo aggregation (2 × 100%) increased cleavage (91.5% vs. 74.4%) and blastocyst (46.0% vs. 31.3%) rates compared to controls (1 × 100%), but did not improve the overall embryo production efficiency on Day 7 (23.0% vs. 31.3%), on a per reconstructed embryo basis, respectively. Treatments had no effects on in vitro developmental kinetics, embryo quality, and in vivo development. In summary, the fusion-activation interval and/or the aggregation of cloned bovine embryos did not affect cloning efficiency based on the in vitro development to the blastocyst stage and on pregnancy outcome.

Impact of cumulative gain in expertise on the efficiency of handmade cloning in cattle.

The aim of this study was to determine the effects of the cumulative gain in expertise in carrying out handmade cloning (HMC) procedures on embryo yield and pregnancy outcome in cattle. Results from in vitro and in vivo embryo development after HMC during three periods of 7 months, separated by 3-month intervals, were compiled and designated as P1, P2 and P3. Blastocyst yield, morphological quality and stage of development, and pregnancy per embryo transfer (ET) on Day 30 of gestation were compared. Zona-intact oocytes were activated chemically in each experiment replicate, and development of parthenogenetic blastocysts was used as a control measurement of oocyte quality and in vitro culture conditions. A total of 21,231 cumulus-oocyte complexes (COCs) were in vitro-matured, with 5,432, 10,721 and 5078 COCs used in 16, 18 and 10 replicates for P1, P2 and P3, respectively. Cloned blastocyst yields on Day 7 increased from 15.5% (124/798) in P1 to 21.6% (309/1428) and 36.6% (280/764) in P2 and P3, respectively. No differences were observed in blastocyst development of parthenogenetic embryos, which average 30.0, 37.6, and 36.4% in P1, P2, and P3, respectively. A 10-fold higher probability of obtaining cloned blastocysts at more advanced stages of development and of higher morphological grade was seen during P3 compared with P1. Pregnancy per ET on Day 30 also increased with gain in expertise, being 6.7% (2/30), 20.8% (10/48) and 40.0% (24/60) for P1, P2 and P3, respectively. The relative efficiency for the establishment of pregnancies (per total COC) increased from 0.04% (1:2716) in P1 to 0.22% (1:460) in P2, reaching 0.47% (1:212) in P3. Results demonstrated a gradual improvement in in vitro and in vivo embryo development over time after establishment of HMC procedures in the laboratory, highlighting the importance of gaining experience and technical skills on the overall cloning efficiency.

Developmental Outcome and Related Abnormalities in Goats: Comparison Between Somatic Cell Nuclear Transfer- and In Vivo-Derived Concepti During Pregnancy Through Term.

Cloning by somatic cell nuclear transfer (SCNT) is characterized by low efficiency and the occurrence of developmental abnormalities, which are rather poorly studied phenomena in goats. This study aimed at comparing overall SCNT efficiency in goats by using in vitro-matured (IVM) or in vivo-matured oocytes and fibroblast donor cells (mock transfected, transgenic, or wild type), also characterizing symptoms of the Abnormal Offspring Syndrome (AOS) in development, comparing results with pregnancies produced by artificial insemination (AI) and in vivo-derived (IVD) embryos. The SCNT group had lower pregnancy rate (18.3%, 11/60), total number of concepti (20.0%, 12/60), term births (3.3%, 2/60), and live births (1.7%, 1/60) than both the IVD (77.8%, 7/9; 155.5%, 14/9; 122.2%, 11/9; 88.8%, 8/9) and the AI (71.4%, 10/14; 121.4%, 17/14; 100%, 14/14; 78.5%, 11/14) groups, respectively (p < 0.05). No SCNT pregnancies reached term using IVM oocytes, but in vivo-matured oocytes resulted in two term transgenic cloned kids. The proportion fetal membrane (FM) weight/birth weight reflected an increase in FM size and cotyledonary enlargement in clones, for disproportionally bigger newborns in relation to cotyledonary numbers. Overall, goat cloning showed losses and abnormality patterns similar to the AOS in cloned cattle and sheep, which have not been previously well recognized in goats.

The use of a metal container for vitrification of mouse ovaries, as a clinical grade model for human ovarian tissue cryopreservation, after different times and temperatures of transport.

PURPOSE: Cryopreservation of ovarian tissue is paramount for fertility preservation, with important clinical applications, especially for women suffering from an oncological condition. Several cryopreservation methodologies have been tried in search of better outcomes, especially in terms of primor-dial and primary follicles integrity post-cryopreservation. Vitrification has successfully been applied to ovarian tissue using different carriers for tissue exposure to the liquid nitrogen (LN2). METHODS: We developed an enclosed metal vessel, which has the advantage of a faster heat transfer, when in contact with LN2 avoiding at the same time, the direct contact with tissue. Additionally, we assessed the effect of different times and temperatures of transport between the collection of mouse ovaries and the beginning of cryopreservation, on follicular morphology after vitrification. RESULTS: Our results suggest that 37 °C and R.T. help to maintain normal primordial and primary follicle morphology for up to 4 hrs after collection and beginning of vitrification in a metal container. CONCLUSION: These data show that the metal container is an appropriate carrier for mouse ovary vitrification. The rate of morphologically normal primordial follicles up to 4 hrs.

Using ERG inquiry to evaluate otoplasty satisfaction in an otorhinolaryngology medical residency training hospital.

UNLABELLED: The ear deformity surgery intervention impact on psychological and self-esteem aspects, in adults and children, is well documented. Recently, the studys are focused on patient satisfaction, funcional result and impact on quality of life. Any modification on patient's quality of life has been a challenge. The use of valid and established questionnairies, like Glasgow Benefit Inventory (GBI), assists on data analyse, turning it consistent. AIM: The aim of this study is to evaluate the impact on patients quality of life after otoplasty, through the GBI questionnaire. METHODS: Retrospective study including patients underwent otoplasty, within july of 2009 to july of 2010. The data were collected through questionnaire applied by medical resident on 90 post-surgical return. RESULTS: 36 patients answered the questionnaire. There was increase on patients quality of life demonstrated by positive mediana obtained through out questinnaire. There were no significantly differences between age and sex. CONCLUSION: The patients are satisfied with post-surgical results. There was increase on patients quality of life conform positive results obtained. The use of GBI showed easy and elucidative.

Utilização do questionário ERG na avaliação do grau de satisfação dos pacientes submetidos à otoplastia em um serviço de residência médica de otorrinolaringologia / Using ERG inquiry to evaluate otoplasty satisfaction in an otorhinolaryngology medical residency training hospital

A importância da intervenção cirúrgica em diminuir o sofrimento psicológico e melhorar a autoestima em pacientes com deformidades auriculares já está bem documentada. As pesquisas têm focado nos resultados das terapias com ênfase no paciente, observando principalmente satisfação, resultado funcional e impacto na qualidade de vida. Quantificar alterações na qualidade de vida tem sido um desafio. O uso de inquéritos válidos, a exemplo da Escala de Resultados de Glasgow (ERG), auxiliam na obtenção dos dados. OBJETIVO: Avaliar o impacto na qualidade de vida dos pacientes submetidos à otoplastia realizada em serviço de residência médica, utilizando como base a ERG, bem como sua funcionalidade. CASUÍSTICA E MÉTODOS: Estudo retrospectivo incluindo pacientes submetidos à otoplastia entre julho de 2009 e julho de 2010. Os dados foram coletados por meio de questionário oferecido ao paciente no retorno pós-operatório. RESULTADOS: Trinta e seis pacientes responderam ao questionário. Houve aumento na qualidade de vida, demonstrado pelas medianas positivas obtidas pelo questionário. Não houve diferença significativa quanto aos valores obtidos entre os sexos e entre diferentes faixas etárias. CONCLUSÃO: Os pacientes mostraram-se satisfeitos com o resultado pós-operatório. Houve aumento de qualidade de vida, conforme demonstrado pelos resultados positivos. A ERG pareceu-nos fácil e elucidativa.

The ear deformity surgery intervention impact on psychological and self-esteem aspects, in adults and children, is well documented. Recently, the studys are focused on patient satisfaction, funcional result and impact on quality of life. Any modification on patient's quality of life has been a challenge. The use of valid and established questionnairies, like Glasgow Benefit Inventory (GBI), assists on data analyse, turning it consistent. AIM: The aim of this study is to evaluate the impact on patients quality of life after otoplasty, through the GBI questionnaire. METHODS: Retrospective study including patients underwent otoplasty, within july of 2009 to july of 2010. The data were collected through questionnaire applied by medical resident on 90 post-surgical return. RESULTS: 36 patients answered the questionnaire. There was increase on patients quality of life demonstrated by positive mediana obtained through out questinnaire. There were no significantly differences between age and sex. CONCLUSION: The patients are satisfied with post-surgical results. There was increase on patients quality of life conform positive results obtained. The use of GBI showed easy and elucidative.

Survival rates of mouse blastocyst vitrified in dimethylformamide based solutions associated with ethylene glicol or 1-2 propanediol / Sobrevivência de embriões de camundongo vitrificados em soluções com dimetilformamida associadas ao etilenoglicol ou 1-2 propanediol

The aim of this study was to determine the effect of dimethylformamide (DF) associated with ethylene glycol (EG) or 1-2 propanediol (PROH) during vitrification, on the in vitro development of mouse blastocysts. Cryoprotectant toxicity was evaluated exposing embryos into three different equilibrium solutions (ES) composed by DF, EG or PROH mixtures (10 percent v/v of each) in mPBS + 0.5 percent PVA at different interval times (1, 3 and 10min). In a second experiment, embryos were exposed to the same ES (either 1 or 3min), following for the three respectively vitrification solutions (VS) (20 percent v/v of each) for 30s. After 72 hours of in vitro culture, embryo hatching and expansion rates were similar for the ES1 and ES2 equilibration solutions during the time interval of 1 or 3min. However embryos exposed for 10 min to the DF equilibration solutions, had lower survival rates than EG-PROH solution (P<0.01). Furthermore, survival rates for embryos exposed to DF-PROH (ES+VS) were lower than embryos exposed to the other solutions (P<0.01). Blastocyst vitrification was performed with the three ES+VS (for 1min and 30s, respectively), using glass micropipettes (GMP). Survival rates were lower for blastocysts vitrified with DF solutions (3 percent-3/108 and 17.1 percent-19/111) (P<0.01) than with PROH+EG vitrification solutions (69 percent-73/105). In conclusion, DF as a cryoprotectant into vitrification solutions have deleterious effects on the in vitro developmental competence of vitrified mouse blastocysts.

O objetivo deste estudo foi determinar o efeito da dimetilformamida (DF) associada com etileno glycol (EG) ou 1-2 propanediol (PROH) durante a vitrificação, no desenvolvimento in vitro de blastocistos murinos. A toxicidade dos crioprotetores foi avaliada ao expor os embriões as três soluções de equilíbrio (ES) compostas pelas misturas de DF, EG ou PROH (10 por cento v/v de cada) em mPBS + 0,5 por cento PVA, em diferentes intervalos de tempo (1, 3 e 10min). Em um segundo experimento, os embriões foram expostos as mesmas ES (durante 1 e 3min), seguido da exposição as três respectivas soluções de vitrificação (VS) (20 por cento v/v de cada) durante 30seg. Após 72 horas de cultivo in vitro, as taxas de expansão e eclosão dos embriões expostos durante os períodos de 1 e 3min às soluções de equilíbrio ES1 e ES2 foram semelhantes. No entanto, os embriões expostos durante 10min às soluções de equilíbrio com DF apresentaram taxas de sobrevivência inferiores à solução de EG-PROH (P<0,01). Além disso, as taxas de sobrevivência dos embriões expostos à DF-PROH (ES+VS) foram menores que as dos embriões expostos as outras soluções (P<0,01). A vitrificação dos blastocistos foi realizada após a exposição dos embriões nas três ES+VS (por 1min e 30seg, respectivamente), usando micropipetas de vidro (GMP). As taxas de sobrevivência foram menores nos blastocistos vitrificados nas soluções compostas por DF (3 por cento-3/108 e 17,1 por cento-19/111), em relação à solução EG-PROH (69 por cento-73/105) (P<0,01). Em conclusão, a DF adicionada como crioprotetor às soluções de vitrificação apresenta efeitos deletérios na capacidade de desenvolvimento in vitro dos blastocistos murinos vitrificados.

Production of bovine hand-made cloned embryos by zygote-oocyte cytoplasmic hemi-complementation.

The aim of this study was to evaluate the effect of the cytoplast type and activation process on development of cloned embryos. Bovine oocytes (MII) or zygotes at the one-cell stage (IVF) were manually bisected and segregated in MII or IVF hemi-cytoplasts or hemi-karyoplasts. Adult skin cells from a bovine female were used as nucleus donors (SC). Experimental groups were composed of IVF embryos; parthenogenetic embryos; hand-made cloned (HMC) embryos; and reconstructed HMC embryos using IVF hemi-cytoplast + MII hemi-cytoplast + SC (G-I); IVF hemi-cytoplast + IVF hemi-cytoplast + SC (G-II); MII hemi-cytoplast + IVF hemi-karyoplast (G-III); and IVF hemi-cytoplast + IVF hemi-karyoplast (G-IV). Embryos from G-I to G-IV were allocated to subgroups as sperm-activated (SA) or were further chemically activated (SA + CA). Embryos from all groups and subgroups were in vitro cultured in the WOW system. Blastocyst development in subgroup G-I SA (28.2%) was similar to IVF (27.0%) and HMC (31.4%) controls, perhaps due to a to a more suitable activation process and/or better complementation of cytoplasmic reprogramming factors, with the other groups and subgroups having lower levels of development. No blastocyst development was observed when using IVF hemi-karyoplasts (G-III and G-IV), possibly due to the manipulation process during a sensitive biological period. In summary, the presence of cytoplasmic factors from MII hemi-oocytes and the sperm activation process from hemi-zygotes appear to be necessary for adequate in vitro development, as only the zygote-oocyte hemi-complementation was as efficient as controls for the generation of bovine cloned blastocysts.

Prevalência de coinfecção tuberculose/HIV em pacientes do Centro de Saúde Modelo de Porto Alegre, Rio Grande do Sul / Prevalence of tuberculosis/HIV coinfection in patients from Model Health Centre in Porto Alegre, Rio Grande do Sul

Objetivos: o estudo teve como propósito estimar a prevalência da infecção pelo Vírus da Imunodeficiência Humana (HIV) nos pacientes em tratamento para tuberculose, no Centro de Saúde Modelo de Porto Alegre, RS.Métodos: foi realizada uma investigação transversal cuja população constituiu-se na totalidade dos pacientes em tratamento para tuberculose no Centro de Saúde Modelo de Porto Alegre, RS, entre 2004 e 2007. Para análise estatística foram utilizados o teste qui-quadrado e análise multivariada pelo método forward likelihood ratio. Resultados: no período em estudo foram diagnosticados 1537 casos de tuberculose, sendo que 449 apresentaram sorologia positiva para o HIV, o que representou uma taxa de coinfecção de 29,2%. Nesse grupo houve predominância do sexo masculino (73,9%) e da faixa etária de 30 a 39 anos (40,8%). No diagnóstico dos coinfectados com HIV/tuberculose a forma clínica prevalente foi a extrapulmonar (49%). Na baciloscopia, a forma negativa prevaleceu (29,8%) e a radiologia sugestiva (74,9%) também predominou nesse grupo. O esquema medicamentoso rifampicina, isoniazida e pirazinamida foi o tipo de tratamento mais usado (87,5%) e a alta por cura nos pacientes com sorologia positiva para o HIV em tratamento para tuberculose foi de 43,7%.Conclusões: os resultados ressaltam a importância da realização do teste sorológico para HIV quando se diagnostica a tuberculose.

Aims: The study aimed to estimate the prevalence of Human Immunodeficiency Virus (HIV) infection in patients undergoing treatment for tuberculosis, at the Model Health Center in Porto Alegre, RS.Methods: This was a cross-sectional study in which the studied population consisted of all patients undergoing treatment for tuberculosis in the Model Health Center in Porto Alegre, RS, from 2004 to 2007. Statistical analysis was made by the chi-square test and by multivariate analysis by the forward likelihood ratio method. Results: In the studied period, 1537 patients were diagnosed with tuberculosis, of whom 449 presented positive serology for HIV, resulting in a rate of coinfection of 29.2%. This group was predominantly male (73.9%), aging 30 to 39 years (40.8%). In cases of HIV/tuberculosis coinfection, the most prevalent clinical form was the extra pulmonary (49%). In bacilloscopy, the negative form prevailed (29.8%), and suggestive radiology (74.9%) also predominated in this group. The therapeutic schedule rifampin, isoniazid and pyrazinamide was the most used (87.5%), and the discharge by cure of treated tuberculosis of patients with positive serology for HIV was 43.7%.Conclusion: The results have emphasized the importance of the serological test for HIV when tuberculosis is diagnosed.

Developmental potential of bovine hand-made clone embryos reconstructed by aggregation or fusion with distinct cytoplasmic volumes.

Animal cloning has been associated with developmental abnormalities, with the level of heteroplasmy caused by the procedure being one of its potential limiting factors. The aim of this study was to determine the effect of the fusion of hemicytoplasts or aggregation of hemiembryos, varying the final cytoplasmic volume, on development and cell density of embryos produced by hand-made cloning (HMC), parthenogenesis or by in vitro fertilization (IVF). One or two enucleated hemicytoplasts were paired and fused with one skin somatic cell. Activated clone and zona-free parthenote embryos and hemiembryos were in vitro cultured in the well-of-the-well (WOW) system, being allocated to one of six experimental groups, on a per WOW basis: single clone or parthenote hemiembryos (1 x 50%); aggregation of two (2 x 50%), three (3 x 50%), or four (4 x 50%) clone or parthenote hemiembryos; single clone or parthenote embryos (1 x 100%); or aggregation of two clone or parthenote embryos (2 x 100%). Control zona-intact parthenote or IVF embryos were in vitro cultured in four-well dishes. Results indicated that the increase in the number of aggregated structures within each WOW was followed by a linear increase in cleavage, blastocyst rate, and cell density. The increase in cytoplasmic volume, either by fusion or by aggregation, had a positive effect on embryo development, supporting the establishment of pregnancies and the birth of a viable clone calf after transfer to recipients. However, embryo aggregation did not improve development on a hemicytoplast basis, except for the aggregation of two clone embryos.

Can electrons travel through actin microfilaments and generate oxidative stress in retinol treated Sertoli cell?

In early reports our research group has demonstrated that 7 microM retinol (vitamin A) treatment leads to many changes in Sertoli cell metabolism, such as up-regulation of antioxidant enzyme activities, increase in damage to biomolecules, abnormal cellular division, pre-neoplasic transformation, and cytoskeleton conformational changes. These effects were observed to be dependent on the production of reactive oxygen species (ROS), suggesting extra-nuclear (non-genomic) effects of retinol metabolism. Besides 7 microM retinol treatment causing oxidative stress, we have demonstrated that changes observed in cytoskeleton of Sertoli cells under these conditions were protective, and seem to be an adaptive phenomenon against a pro-oxidant environment resulting from retinol treatment. We have hypothesized that the cytoskeleton can conduct electrons through actin microfilaments, which would be a natural process necessary for cell homeostasis. In the present study we demonstrate results correlating retinol metabolism, actin architecture, mitochondria physiology and ROS, in order to demonstrate that the electron conduction through actin microfilaments might explain our results. We believe that electrons produced by retinol metabolism are dislocated through actin microfilaments to mitochondria, and are transferred to electron transport chain to produce water. When mitochondria capacity to receive electrons is overloaded, superoxide radical production is increased and the oxidative stress process starts. Our results suggested that actin cytoskeleton is essential to oxidative stress production induced by retinol treatment, and electrons conduction through actin microfilaments can be the key of this correlation.