Assessment of ALA-induced PpIX production in porcine skin pretreated with microneedles

Photodynamic therapy (PDT) is used for skin treatments of premalignant and cancer lesions and recognized as a non-invasive technique that combines tissue photosensitization and subsequent exposure to light to induce cell death. However, it is limited to the treatment of superficial lesions, mainly due to the low cream penetration. Therefore, the improvement of transdermal distribution of aminolevulinic acid (ALA) is needed. In this study, the kinetics and homogeneity of production of ALA-induced PpIX after the skin pre-treatment with microneedles rollers of 0.5, 1.0 and 1.5 mm length were investigated. An improvement in homogeneity and production of PpIX was shown in a porcine model. Widefield fluorescence imaging three hours after the topical application of ALA-cream in the combined treatment with microeedles rollers.

Concentrações séricas hormonais em vacas azebuadas submetidas à baixa e alta ingestão alimentar

O objetivo deste trabalho foi avaliar a influência da ingestão alimentar nas concentrações séricas de hormônios reprodutivos e metabólicos em vacas azebuadas. Dezoito vacas foram divididas em dois grupos: 170% (alta ingestão = A) e 66% (baixa ingestão = B) da dieta de manutenção. Com 21 dias nas dietas experimentais, as vacas tiveram o estro sincronizado. Posteriormente, os ovários foram avaliados por ultra-sonografia transretal e sangue foi coletado diariamente até o dia 7 do ciclo (ovulação = dia 1). Na análise estatística, utilizou-se o teste t. As vacas ganharam 1,1 kg por dia no grupo A e perderam 1,5 kg por dia de PV no grupo B. Apesar de não ter havido diferença entre os grupos no diâmetro máximo do folículo ovulatório, o grupo A apresentou pico pré-ovulatório de estradiol sérico menor. Não foi observada diferença entre os grupos quanto ao volume luteal e concentração sérica de progesterona no dia 7 do ciclo e de FSH, IGF-I e insulina séricos no período peri-ovulatório. As dietas experimentais não alteraram a função ovariana e as concentrações séricas de hormônios reprodutivos e metabólicos, com exceção do estradiol, sugerindo que, no grupo de alta ingestão, ocorreu maior metabolismo desse hormônio.

Comparison of three protocols for superovulation of brown brocket deer (Mazama gouazoubira)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção in vitro de embriões bovinos: utilização de diferentes fontes de gonadotrofinas na maturação dos oócitos

O presente trabalho foi conduzido com o objetivo de avaliar o efeito da utilização de diferentes fontes de gonadotrofinas para maturação in vitro dos oócitos bovinos fecundados e desenvolvidos in vitro sobre as taxas de clivagem (TC) e de blastocistos (TBL). Oócitos imaturos provenientes de ovários de vacas de abatedouro foram submetidos a maturação in vitro sob diferentes condições: meio TCM 199, acrescido de 10% de soro de vaca em estro (SVE), aditivos, hepes, NaHCO3, piruvato de sódio, antibióticos (meio B-199), 20 UI/mL de PMSG e 10 UI/mL de hCG (PMSG/hCG) ou meio B-199, acrescido de 5 mig/mL de FSH e 5 mig/mL de LH (FSH/LH). Seguidos 24 h de cultura a 38,5ºC em atmosfera com 5% de CO2, os oócitos maturos foram incubados com sêmen descongelado durante 18 a 21 horas. Após esse período, os oócitos foram transferidos para placas contendo microgotas de meio Ménezo suplementado com 10% de SVE e células epiteliais do oviduto bovino em suspensão, cobertas com óleo de silicone, os quais permaneceram em cultura por mais 9 dias. Os dados foram analisados pelo teste do Qui-quadrado. A TC e a TBL, para PMSG/hCG e FSH/LH, foram 60 e 13,9% e 61,2 e 10,6%, respectivamente. Não houve diferença entre os tratamentos com relação a TC ou a TBL. Esses resultados sugerem que ambas as fontes de gonadotrofinas podem ser utilizadas para maturação in vitro dos oócitos fecundados e desenvolvidos in vitro.

In Vitro Ultrasound Influence on Cutaneous Permeation of Hyaluronidase

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo mecânico da estabilidade dorsal da osteossíntese intramedular associada à fixação externa (método de Ulson)

OBJETIVO: Avaliar o método de Ulson de fixação intramedular associado à fixação externa variando a altura do travamento externo dos fios de Kirschner e sem fixação externa. MÉTODO: Foram utilizadas 18 tíbias de porcos, sendo realizada osteotomia transversal na região da tuberosidade e introduzidos dois fios de Kirschner intramedulares em cada peça, em três diferentes padrões de montagem: grupo I - travamento com minifixador externo com 3,0cm de altura; grupo II - travamento com 4,5cm de altura; grupo III: sem travamento externo. Realizaram-se ensaios mecânicos de cisalhamento, obtendo-se: carga máxima, limite de proporcionalidade e coeficiente de rigidez. RESULTADOS: Não foram encontradas diferenças significativas de carga máxima e limite de proporcionalidade entre os grupos; o grupo II apresentou maior coeficiente de rigidez. CONCLUSÃO: A altura do travamento dos fios de Kirschner no método de Ulson, dentro dos limites avaliados, não prejudicou a estabilidade do sistema de fixação da fratura.

Perfil dos hormônios da tireoide em pacientes com câncer de mama em estado de menopausa

Objective: The aim of this study was to determine thyroid hormone (TH) profile in postmenopausal patients with breast cancer (BC). Subjects and methods: 12 CaM patients stages I or II, without interventions that could interfere with tumor progression were selected, as well as and a control group with 18 postmenopausal women without CaM. We measured serum anti-thyroperoxidase antibody (TPOAB), thyroid-stimulating hormone (TSH), free thyroxine (T4L), estradiol (E2), follicle-stimulating hormone (FSH), and luteinizing hormone (LH), before and after surgery, besides immunohistochemistry for estrogen (ER) and progesterone (PR) receptors. Results: Four patients with CaM showed changes in thyroid hormone profile: two had hyperthyroidism, one hypothyroidism, and one was positive for TPO-AB. All of them positive for ER and PR.TSH levels in breast cancer patients were not different from levels found in the control group (1.89 +/- 1.56 vs. 2.86 +/- 3.12 mIU/mL), but the levels of T4L in patients with CaM were statistically higher than those of the control group (1.83 +/- 0.57 vs. 1.10 +/- 0.20 ng/dL). Conclusion: These results reinforce the need for assessment of thyroid status in CaM patients, since in the absence of E2, changes in clinical HTs can act in E2-controlled processes. Arq Bras Endocrinol Metab. 2012;56(4):238-43

Recombinant luteinizing hormone supplementation to recombinant follicle-stimulation hormone during induced ovarian stimulation in the GnRH-agonist protocol: A meta-analysis

Purpose: to compare the efficacy of recombinant LH supplementation for controlled ovarian stimulation in recombinant FSH and GnRH-agonist protocol.Methods: Search strategies included on-line surveys of databases. The fixed effects model was used for odds ratio and effect size (weighted mean difference). Four trials fulfilled the inclusion criteria.Results: a fewer days of stimulation (p < 0.0001), a fewer total amount of r-FSH administered (p < 0.0001) and a higher serum estradiol levels on the day of hCG administration (p < 0.0001) were observed for the r-LH supplementation protocol. However, differences were not observed in number of oocyte retrieved, number of mature oocytes, clinical pregnancy per oocyte retrieval, implantation and miscarriage rates.Conclusions: more randomized controlled trials are necessary before evidence-based recommendations regarding exogenous LH supplementation in ovarian stimulation protocols with FSH and GnRH-agonist for assisted reproduction treatment can be provided.

Plasma hormones, muscle mass and strength in resistance-trained postmenopausal women

Objective: To associate changes of body composition, muscle strength (MS) and plasma hormones (PH) in resistance-training protocol in sedentary postmenopausal women (PMW).Design: This randomized controlled trial, Brazilian 43 PMW (45-70-year-old) able for physical exercises were selected after they have accomplished medical and ethical criteria. They were assigned in two groups: RT, resistance training (n = 22); and CT, not trained control (n = 2 1); with supervision sessions of two to three exercise for large and one exercise for smaller groups in three series of 8-12 rep. (60-80% 1RM) for each exercise. The training period lasted 16 weeks and was preceded by low-load exercise (40-50% 1RM) adaptation period of 4 weeks (3/(times week)). Body weight, height, body mass index (BMI), and composition (BIA) along with fast-PH (FSH, LH, estrachol, cortisol, IGF-1 and testosterone) were assessed before (MO) and after (M 16) the 4 weeks period with the MS (1RM) determined also at 8 weeks (W). The values were correlated by Person's test and the means compared by Student's t-test and ANOVA.Results: At baseline both groups were similar in age, time of PMW, body composition, MS and fast-PH. However after 16 weeks, RT presented higher BMI (2. 1 %), IGF- 1 (37.8%) and MM gain (1.8 +/- 0.8 kg) than CT. MM correlated positively with IGF-1 (r = 0.45, p < 0.05) and MS progressively increased in all exercise greater in pectoral than legs and upper arms.Conclusion: Former sedentary postmenopausal women submitted to resistance training gained MM and MS irrespectively of fat mass changes but significantly associated with IGF-1 increase. (C) 2008 Elsevier B.V. All rights reserved.

Adjuvant therapy with GnRH agonists/tamoxifen in breast cancer should be a good council for patients with hormone receptor-positive tumours and wish to preserve fertility

Infertility represents one of the main long-term consequences of the chemotherapy used for the adjuvant treatment of breast cancer. Approximately 60-65% of breast cancers express the nuclear hormone receptor in premenopausal women. Adjuvant endocrine therapy is an integral component of care for patients with hormone receptor-positive (HR+) tumours. The GnRH agonist (GnRHa) alone or in combination with tamoxifen produces results at least similar to those obtained with the different chemotherapy protocols in patients with HR+ breast cancer with respect to recurrence-free survival and overall survival. It is time to indicate adjuvant therapy with GnRHa associated with tamoxifen for patients with breast cancer (HR+ tumours) if they want to preserve their reproductive function. The evaluation of ovarian reserve tests: follicle stimulating hormone (FSH), anti-Mullerian hormone (AMH), inhibin B, antral follicle count (AFC) and ovarian volume 6 months, and 1 year after the end of therapy with GnRHa/tamoxifen must be realised. The recurrence-free survival and overall survival should be analysed. The major implication of this hypothesis will be to avoid adjuvant chemotherapy for patients with breast cancer (HR+ tumours) that request fertility preservation. It is expected that ovarian function should not be altered in almost all cases and subsequent pregnancy a real possibility. (C) 2012 Elsevier Ltd. All rights reserved.

GnRH agonist versus GnRH antagonist in IVF/ICSI cycles with recombinant LH supplementation: DNA fragmentation and apoptosis in granulosa cells

Objective: To compare the level of apoptosis and DNA fragmentation in the human granulosa cell (GC) layer exposed to an agonist or antagonist of GnRH in intracytoplasmic sperm injection (ICSI) cycles supplemented with recombinant LH (rLH).Study design: Patients without ovulatory dysfunction, aged <= 37 years and in their first ICSI cycle were prospectively randomised to receive either a long GnRH agonist protocol or a multi-dose antagonist protocol. In both groups, recombinant FSH supplemented with rLH was used for ovarian stimulation, and the GCs were collected during oocyte denudation. The GCs were then analysed for DNA fragmentation by TUNEL assay and for apoptosis using the annexin-V assay. The outcomes were given as the percentage of GCs with DNA fragmentation and apoptosis out of the total number of GCs analysed. Comparison of the agonist versus the antagonist group was performed using the Mann-Whitney test.Results: DNA fragmentation: 32 patients were included in either the GnRH agonist group (n = 16) or the antagonist group (n = 16). The percentage of GCs with positive DNA fragmentation did not differ significantly (P = 0.76) between the agonist group (15.5 +/- 9.4%) and the antagonist group (18.8 +/- 13.3%). Apoptosis: 28 patients were included in either the GnRH agonist group (n = 14) or the antagonist group (n = 14). The percentage of GCs positive for apoptosis did not differ significantly (P = 0.78) between the agonist group (34.6 +/- 14.7%) and the antagonist group (36.5 +/- 22%).Conclusions: The results suggest that therapy with either an agonist or antagonist of GnRH is associated with comparable levels of DNA fragmentation and apoptosis in granulosa cells in ICSI cycles supplemented with rLH. (C) 2012 Elsevier B.V. All rights reserved.

Vitamin C partially attenuates male reproductive deficits in hyperglycemic rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efficacy of the motile sperm organelle morphology examination (MSOME) in predicting pregnancy after intrauterine insemination

Background: Although the motile sperm organelle morphology examination (MSOME) was developed merely as a selection criterion, its application as a method for classifying sperm morphology may represent an improvement in the evaluation of semen quality. The aim of this study was to determine the prognostic value of normal sperm morphology using MSOME with regard to clinical pregnancy (CP) after intrauterine insemination (IUI).Methods: A total of 156 IUI cycles that were performed in 111 couples were prospectively analysed. Each subject received 75 IU of recombinant FSH every second day from the third day of the cycle. Beginning on the 10th day of the cycle, follicular development was monitored by vaginal ultrasound. When one or two follicles measuring at least 17 mm were observed, recombinant hCG was administered, and IUI was performed 12-14 h and 36-40 h after hCG treatment. Prior to the IUI procedure, sperm samples were analysed by MSOME at 8400x magnification using an inverted microscope that was equipped with DIC/Nomarski differential interference contrast optics. A minimum of 200 motile spermatozoa per semen sample were evaluated, and the percentage of normal spermatozoa in each sample was determined.Results: Pregnancy occurred in 34 IUI cycles (CP rate per cycle: 21.8%, per patient: 30.6%). Based on the MSOME criteria, a significantly higher percentage of normal spermatozoa was found in the group of men in which the IUI cycles resulted in pregnancy (2.6+/-3.1%) compared to the group that did not achieve pregnancy (1.2+/-1.7%; P = 0.019). Logistic regression showed that the percentage of normal cells in the MSOME was a determining factor for the likelihood of clinical pregnancy (OR: 1.28; 95% CI: 1.08 to 1.51; P = 0.003). The ROC curve revealed an area under the curve of 0.63 and an optimum cut-off point of 2% of normal sperm morphology. At this cut-off threshold, using the percentage of normal sperm morphology by MSOME to predict pregnancy was 50% sensitive with a 40% positive predictive value and 79% specificity with an 85% negative predictive value. The efficacy of using the percentage of normal sperm morphology by MSOME in predicting pregnancy was 65%.Conclusions: The present findings support the use of high-magnification microscopy both for selecting spermatozoa and as a routine method for analysing semen before performing IUI.

Glutamate-induced obesity leads to decreased sperm reserves and acceleration of transit time in the epididymis of adult male rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeitos da isoflavona sobre os sintomas climatéricos e o perfil lipídico na mulher em menopausa

OBJETIVO: avaliar os efeitos da isoflavona, do gérmen da soja, sobre os sintomas climatéricos e o perfil lipídico na mulher em menopausa. MÉTODOS: foi conduzido estudo prospectivo, com 50 mulheres em menopausa, divididas em: G1, usuárias de isoflavona (60 mg/dia) (n=25), e G2, placebo (n=25). Os critérios de inclusão foram FSH >40 mUI/mL e presença de fogachos. Foram excluídas as vegetarianas, fumantes, asiáticas, portadoras de doenças gastrointestinais e usuárias de terapia de reposição hormonal. No seguimento, de seis meses, foram obtidos o índice menopausal de Kupperman (IMK), o perfil hormonal e o lipídico. Na análise estatística, empregaram-se ANOVA, o teste t pareado e as provas não paramétricas de Wilcoxon e Mann-Whitney. RESULTADOS: os valores medianos do IMK, inicialmente iguais entre os grupos (IMK = 20), reduziram-se nas usuárias de isoflavona aos 2 e 4 meses (IMK = 14 e 9, respectivamente) e no grupo controle, apenas aos 2 meses (IMK = 15) (p<0,01). Ao final do estudo, a isoflavona foi superior ao placebo na redução dos fogachos (44% versus 12%, respectivamente). Aos seis meses, verificou-se que os valores médios de estradiol foram superiores no G1 quando comparados ao G2 (18,0 ± 6,7 versus 12,3 ± 3,8 ng/dL) (p<0,05), sem alterações no FSH e LH. Entre as usuárias de isoflavona, houve redução de 11,8% no LDL (de 151,5 ± 39,2 para 133,6 ± 26,4 mg/dL) e elevação de 27,3% no HDL (de 44,0 ± 11,3 para 56,0 ± 11,9 mg/dL) (p<0,05). CONCLUSÕES: a isoflavona, do gérmen da soja, induziu efeitos favoráveis sobre os sintomas climatéricos e o perfil lipídico, revelando-se opção interessante como terapêutica alternativa para mulheres em menopausa.