Disposable Bioreactors Used in Process Development and Production Processes with Plant Cell and Tissue Cultures.

The bioreactor is the centerpiece of the upstream processing in any biotechnological production process. Its design, the cultivation parameters, the production cell line, and the culture medium all have a major influence on the efficiency of the process and the result of the cultivation. Disposable bioreactors have been used for the past 20 years, playing a major role in process development and commercial production of high-value substances at medium scales.Our review deals with scalable, disposable bioreactors that have proven to be useful for the cultivation of plant cell and tissue cultures. Based on the definitions of terms and a categorization approach, the most commonly used, commercially available, disposable bioreactor types are presented below. The focus is on wave-mixed, stirred, and orbitally shaken bioreactors. In addition to their instrumentation and bioengineering characteristics, cultivation results are discussed, and emerging trends for the development of disposable bioreactors for plant cell and tissue cultures are also addressed.

Factors related with osteoporosis treatment in postmenopausal women.

Although fractures had high mortality and morbidity, many studies proved that fracture risk might be decreased by pharmacological therapy, although a low treatment adherence rate is observed. The aim of this study was to identify factors associated with osteoporosis treatment in postmenopausal women.A cross-sectional study was carried out from March to August 2013 at the primary care setting. Postmenopausal women were recruited. A standardized questionnaire was applied. Women who were using at least one of the following drugs at the moment of the survey were considered as current treatment: bisphosphonates, raloxifene, estrogen, calcitonin, teriparatide, or strontium ranelate. Women who had used any of the mentioned medications before the study were considered as past treatment.Of the 1025 women included in the study, 8% were on current treatment, 5.7% had past treatment, and 86.3% had not received treatment. Treated women (either current or past) had a higher rate of osteoarthritis, had more falls, had higher education level, presented a higher rate of private health insurance, and received more information about osteoporosis. They also had more dual-energy x-ray absorptiometry (DXA) scans and were more frequently diagnosed with osteoporosis by these DXA scans. The factors independently associated with treatment in the regression analysis were the DXA scan itself, the diagnosis of osteoporosis by DXA, and information about osteoporosis.Current and past treatments of osteoporosis were associated with DXA and information. These results suggest that some measures to inform women about osteoporosis and or even the popularization of DXA scans could improve the treatment.

Prevalence of human papillomavirus types and variants and p16(INK4a) expression in head and neck squamous cells carcinomas in São Paulo, Brazil.

BACKGROUND: Human papillomavirus (HPV) prevalence in head and neck squamous cell carcinomas (HNSCC) diverges geographically. The reliability of using p16(INK4a) expression as a marker of viral infection is controversial in HNSCC. We evaluated HPV types and HPV-16 variants prevalence, and p16(INK4a) expression in HNSCC specimens provided by two different Institutions in São Paulo. METHODS: HPV DNA from formalin-fixed specimens was accessed by Inno-LiPA, HPV-16 variants by PCR-sequencing, and p16(INK4a) protein levels by immunohistochemistry. RESULTS: Overall, HPV DNA was detected among 19.4 % of the specimens (36/186). Viral prevalence was higher in the oral cavity (25.0 %, 23/92) then in other anatomical sites (oropharynx 14,3 %, larynx 13.7 %) when samples from both Institutions were analyzed together. HPV prevalence was also higher in the oral cavity when samples from both Institutions were analyzed separately. HPV-16 was the most prevalent type identified in 69.5 % of the HPV positive smaples and specimens were assigned into Asian-American (57.2 %) or European (42.8 %) phylogenetic branches. High expression of p16(INK4a) was more common among HPV positive tumors. CONCLUSION: Our results support a role for HPV-16 in a subset of HNSCC.

Comparative study between biopsy and brushing sampling methods for detection of human papillomavirus in oral and oropharyngeal cavity lesions, / Estudo comparativo entre biópsia e escovado na pesquisa do papilomavírus humano em lesões de cavidade oral e de orofaringe

ABSTRACT INTRODUCTION: Many epidemiological studies have suggested that human papillomavirus (HPV), especially type 16, is involved in the genesis of squamous cell carcinoma of the oral cavity and oropharynx, especially in young, non-smoking patients; thus, its detection in lesions in this region is important. OBJECTIVE: To clarify the capacity of the brushing sampling method to detect the presence of HPV in oral or oropharyngeal lesions through polymerase chain reaction (PCR) testing, and to compare the results with those obtained by biopsy. METHODS: Prospective study of adult patients with oral or oropharyngeal lesions assessed by PCR, comparing biopsy specimens with samples obtained by the brushing method. The study was approved by the Research Ethics Committee of the institution. RESULTS: A total of 35 sample pairs were analyzed, but 45.7% of the brushing samples were inadequate (16/35) and, thus, only 19 pairs could be compared. There was agreement of results in 94.7% (18/19) of the pairs, with HPV identified in 16 of them. HPV DNA was detected in 8.6% (3/35) of biopsy and 5.7% (2/35) of brushing samples. CONCLUSION: There was no statistically significant difference between the two methods, but the brushing sampling method showed a higher number of inadequate samples, suggesting that it is an unreliable method for surveillance.

Resumo INTRODUÇÃO: Muitos estudos epidemiológicos indicam a participação do papilomavírus humano, especialmente o tipo 16, na carcinogênese dos tumores espinocelulares das cavidade oral e oro-faríngea, principalmente em jovens e não fumantes, sendo portanto importante sua detecção nas lesões desta região. OBJETIVO: Elucidar a habilidade do escovado em detectar o papilomavírus humano, pela reação em cadeia da polimerase, nas lesões orais e orofaríngeas, comparando os resultados com os obtidos por biópsia. MÉTODO: Estudo prospectivo de pacientes com lesões orais e orofaríngeas, pela reação em cadeia da polimerase, no qual foram pareados os resultados de amostras obtidas por escovado e por biópsia. A pesquisa foi aprovada pelo Comitê de Ética em Pesquisa da instituição. RESULTADO: Foram analisados 35 pares de amostras, porém estavam inapropriadas para análise 45,7% (16/35) das amostras obtidas por escovado, e portanto, somente 19 pares puderam ser comparados. Em 94,7% dos pares houve concordância dos resultados, sendo encontrado o papilomavírus humano − 16 em um destes pares. O ácido desoxirribonucleico do papilomavírus humano foi detectado em 8,6% (3/35) das biópsias e em 5,7% (2/35) dos escovados. CONCLUSÃO: Não houve diferença estatística entre os métodos, mas como houve um grande número de amostras obtidas por escovado inapropriadas, este parece não ser confiável para o rastreamento.

Comparative study between biopsy and brushing sampling methods for detection of human papillomavirus in oral and oropharyngeal cavity lesions.

INTRODUCTION: Many epidemiological studies have suggested that human papillomavirus (HPV), especially type 16, is involved in the genesis of squamous cell carcinoma of the oral cavity and oropharynx, especially in young, non-smoking patients; thus, its detection in lesions in this region is important. OBJECTIVE: To clarify the capacity of the brushing sampling method to detect the presence of HPV in oral or oropharyngeal lesions through polymerase chain reaction (PCR) testing, and to compare the results with those obtained by biopsy. METHODS: Prospective study of adult patients with oral or oropharyngeal lesions assessed by PCR, comparing biopsy specimens with samples obtained by the brushing method. The study was approved by the Research Ethics Committee of the institution. RESULTS: A total of 35 sample pairs were analyzed, but 45.7% of the brushing samples were inadequate (16/35) and, thus, only 19 pairs could be compared. There was agreement of results in 94.7% (18/19) of the pairs, with HPV identified in 16 of them. HPV DNA was detected in 8.6% (3/35) of biopsy and 5.7% (2/35) of brushing samples. CONCLUSION: There was no statistically significant difference between the two methods, but the brushing sampling method showed a higher number of inadequate samples, suggesting that it is an unreliable method for surveillance.

Influence of estradiol administration on estrogen receptors of nasal mucosa: an experimental study on guinea pigs.

INTRODUCTION: Some clinical trials revealed a correlation between increased serum estrogen and nasal symptoms or inflammatory changes in nasal mucosa. Estrogen receptors tend to be controlled by a negative feedback, to avoid a deleterious stimulus over several body functions while in hyperestrogenic periods. This study proposes a hypothesis where mechanisms regulating expression of estradiol receptors in nasal mucosa are absent in some patients, and their concentration remains steady even in periods of high serum hormonal concentration, potentially leading to local estrogenic symptoms in nasal mucosa. STUDY DESIGN: This was an experimental prospective study. AIM: To determine whether estrogen levels induce the reduction of the number of estrogen receptors in the nasal mucosa. METHODS: In the present study, 30 adult male guinea pigs were subjected to a biopsy of the middle nasal turbinate and received 0.5 mL of estradiol cypionate intraperitoneally for 30 consecutive days. Afterwards, samples from contralateral middle turbinate were obtained. Immunohistochemical analysis of estrogen receptors were performed pre- and post-treatment. RESULTS: The post-treatment group showed reduction of receptor expression when compared to the pre-treatment group. (p = 5.2726-5). CONCLUSION: A reduction in the expression of the nasal estrogen receptor was observed after 30 days of estradiol administration.

A influencia da administracao de estradiol nos receptores de estrogeno da mucosa nasal: estudo experimental em cobaias / Influence of estradiol administration on estrogen receptors of nasal mucosa: an experimental study on guinea pigs

Introdução: A literatura indica uma correlação entre estrogênio elevado no soro e sintomas nasais ou alterações inflamatórias na mucosa nasal. Os receptores de estrogênio tendem a ser controlados por retroalimentação negativa, para evitar um estímulo nocivo sobre as diversas funções corporais em períodos de hiperestrogenismo. Propomos uma hipótese em que os mecanismos que regulam a expressão de receptores de estradiol na mucosa nasal estão ausentes em alguns pacientes, e a sua concentração permanece estável mesmo em períodos de elevada concentração sérica hormonal, o que pode conduzir a sintomas locais na mucosa nasal. Desenho do estudo: Estudo prospectivo experimental. Objetivo: Determinar se altos níveis de estrogênio induzem à redução no número de receptores de estrogênio na mucosa nasal. Material e método: Trinta cobaias foram submetidas à biópsia da concha nasal, recebendo 0,5 ml de cipionato de estradiol por via intraperitoneal por trinta dias consecutivos. Em seguida foram obtidas amostras da concha nasal contralateral. As análises imuno-histoquímicas dos receptores de estrógeno foram realizadas antes e depois da hormonioterapia. Resultados: O grupo pós-tratamento mostrou uma redução da expressão dos receptores (p = 5,2726-5). Conclusão: Redução na expressão do receptor de estrogênio nasal foi encontrada após trinta dias de administração de estradiol. .

Introduction: Some clinical trials revealed a correlation between increased serum estrogen and nasal symptoms or inflammatory changes in nasal mucosa. Estrogen receptors tend to be controlled by a negative feedback, to avoid a deleterious stimulus over several body functions while in hyperestrogenic periods. This study proposes a hypothesis where mechanisms regulating expression of estradiol receptors in nasal mucosa are absent in some patients, and their concentration remains steady even in periods of high serum hormonal concentration, potentially leading to local estrogenic symptoms in nasal mucosa. Study design: This was an experimental prospective study. Aim: To determine whether estrogen levels induce the reduction of the number of estrogen receptors in the nasal mucosa. Methods: In the present study, 30 adult male guinea pigs were subjected to a biopsy of the middle nasal turbinate and received 0.5 mL of estradiol cypionate intraperitoneally for 30 consecutive days. Afterwards, samples from contralateral middle turbinate were obtained. Immunohistochemical analysis of estrogen receptors were performed pre- and post-treatment. Results: The post-treatment group showed reduction of receptor expression when compared to the pre-treatment group. (p = 5.2726-5). Conclusion: A reduction in the expression of the nasal estrogen receptor was observed after 30 days of estradiol administration. .

Determination of HPV prevalence in oral/oropharyngeal mucosa samples in a rural district of São Paulo.

UNLABELLED: Knowledge about HPV infection in the oral cavity/oropharynx may contribute to the elucidation of the role it plays in Head and Neck Squamous Cell Carcinoma (HNSCC). OBJECTIVE: To determine the effectiveness of the methodology used for sampling the oral cavity and oropharynx mucosae and to determine the prevalence of HPV in the oral cavity and oropharynx of adults and children. METHOD: The study population was served by an assistance program in a rural district of São Paulo. The subjects were asked to donate samples regardless of complaints. RESULTS AND CONCLUSION: The study included 47 men, 77 women and 22 children, of which the oral cavity samples were obtained by gargling with commercially-available antiseptic mouthwash. We found 3 positive samples (2.4%) in adults: 2 HPV 55 and one HPV 58. No positive results were found in children. Furthermore we concluded that the sampling method with the mouthwash proved effective and fast for the detection of HPV in the oral cavity and oropharynx in the general population.

Determinacao da prevalencia de HPV em amostras de mucosa oral/orofaringea em um distrito rural de Sao Paulo / Determination of HPV prevalence in oral/oropharyngeal mucosa samples in a rural district of Sao Paulo

OBJETIVO: Averiguar a eficácia da metodologia para coleta de amostras em cavidade oral e orofaringe e determinar a prevalência do HPV na cavidade oral e orofaringe de adultos e crianças. MÉTODO: A população estudada foi atendida por um programa assistencial em um distrito rural de São Paulo. Os indivíduos foram convidados a doar amostras independentemente de queixas. RESULTADOS E CONCLUSÃO: Foram incluídos no estudo 47 homens, 77 mulheres e 22 crianças, dos quais amostras da cavidade oral foram obtidas por bochecho e gargarejo com antisséptico oral comercial. Foram encontrados três resultados positivos (2,4%) em adultos, duas amostras de HPV 55 e uma amostra de HPV 58. Não foram observados resultados positivos em crianças. Além disso, concluímos que o método de coleta com o enxágue bucal com antisséptico mostrou-se eficaz e rápido para a detecção de HPV na cavidade oral e orofaríngea na população geral. .

Knowledge about HPV infection in the oral cavity/oropharynx may contribute to the elucidation of the role it plays in Head and Neck Squamous Cell Carcinoma (HNSCC). OBJECTIVE: To determine the effectiveness of the methodology used for sampling the oral cavity and oropharynx mucosae and to determine the prevalence of HPV in the oral cavity and oropharynx of adults and children. METHOD: The study population was served by an assistance program in a rural district of São Paulo. The subjects were asked to donate samples regardless of complaints. RESULTS AND CONCLUSION: The study included 47 men, 77 women and 22 children, of which the oral cavity samples were obtained by gargling with commercially-available antiseptic mouthwash. We found 3 positive samples (2.4%) in adults: 2 HPV 55 and one HPV 58. No positive results were found in children. Furthermore we concluded that the sampling method with the mouthwash proved effective and fast for the detection of HPV in the oral cavity and oropharynx in the general population. .

[Sexuality evaluation in women submitted to hysterectomy for the treatment of uterine leiomyoma]. / Avaliação da sexualidade em mulheres submetidas à histerectomia para tratamento do leiomioma uterino.

PURPOSE: To evaluate the impact of hysterectomy on the sexuality of women with uterine leiomyoma. METHODS: Prospective study including 33 sexually active women, with ages from 35 to 50 years old, with orgasmic experience and with a fit stable partner. All the women were submitted to two instruments for the evaluation or their sexuality: Sexual Quotient - Female Version (SQF) and Sexual Satisfaction Inventory - Female Version (SSIF). Both instruments were applied by the same examiner, before and six months after the hysterectomy. RESULTS: The SQF has shown that 39.4% of the patients presented deterioration in the sexual intercourse, even though there has not been found an association between the SQF results before and after hysterectomy (chi2= 0.6; degree of freedom=12; p=0.05). The mean scores obtained after the application of the SSIF have shown significant deterioration in the following parameters: sexual satisfaction (p=0.03); expression of feminine sensuality (p=0.01); vaginismus/dyspareunia (p=0.02) and an orgasmia (p=0.04). CONCLUSIONS: It seems that hysterectomy has a negative impact on women's sexual life, with reports of decreased libido, arousal and orgasmic capacity.

Avaliação da sexualidade em mulheres submetidas à histerectomia para tratamento do leiomioma uterino / Sexuality evaluation in women submitted to hysterectomy for the treatment of uterine leiomyoma

OBJETIVO: avaliar o impacto da histerectomia sobre a sexualidade de mulheres portadoras de leiomioma uterino. MÉTODOS: estudo prospectivo, no qual foram incluídas 33 mulheres sexualmente ativas, com idade de 35 a 50 anos, experiência orgástica e parceiro fixo capacitado para o coito. Todas as mulheres foram submetidas a dois instrumentos para avaliação da sexualidade: Quociente Sexual - Versão Feminina (QS-F) e Inventário de Satisfação Sexual - Versão Feminina (GRISS). Os mesmos instrumentos foram aplicados pelo mesmo examinador antes da histerectomia e seis meses após o procedimento. RESULTADOS: o QS-F apontou que 39,4 por cento das pacientes apresentaram piora no relacionamento sexual, apesar de não ter sido encontrada associação entre os resultados obtidos no QS-F antes e depois da histerectomia (χ2=10,6; grau de liberdade=12; p=0,05). Os escores médios obtidos após a aplicação do questionário de GRISS mostraram piora significante nos parâmetros "satisfação sexual" (p=0,03); "expressão da sensualidade feminina" (p=0,01); "vaginismo/dispareunia" (p=0,02) e "anorgasmia" (p=0,04). CONCLUSÕES: a histerectomia parece impactar negativamente a vida sexual das mulheres, sendo referida pela diminuição do desejo, da excitação e da capacidade orgásmica.

PURPOSE: to evaluate the impact of hysterectomy on the sexuality of women with uterine leiomyoma. METHODS: prospective study including 33 sexually active women, with ages from 35 to 50 years old, with orgasmic experience and with a fit stable partner. All the women were submitted to two instruments for the evaluation or their sexuality: Sexual Quotient - Female Version (SQF) and Sexual Satisfaction Inventory - Female Version (SSIF). Both instruments were applied by the same examiner, before and six months after the hysterectomy. RESULTS: the SQF has shown that 39.4 percent of the patients presented deterioration in the sexual intercourse, even though there has not been found an association between the SQF results before and after hysterectomy (χ2= 0.6; degree of freedom=12; p=0.05). The mean scores obtained after the application of the SSIF have shown significant deterioration in the following parameters: sexual satisfaction (p=0.03); expression of feminine sensuality (p=0.01); vaginismus/dyspareunia (p=0.02) and anorgasmia (p=0.04). CONCLUSIONS: it seems that hysterectomy has a negative impact on women's sexual life, with reports of decreased libido, arousal and orgasmic capacity.

[Aromatase expression in invasive and in situ ductal carcinoma present in the same breast]. / Expressão da aromatase em carcinomas de mama ductais invasivo e in situ presentes na mesma mama.

OBJECTIVE: to evaluate the expression of aromatase in simultaneously invasive ductal carcinoma (IDC) and ductal carcinoma in situ (DCIS). METHODS: forty-five surgical samples were obtained from mastectomy and quadrantectomy with simultaneous IDC and DCIS of stage I and II patients. Aromatase was evaluated using antibodies anti-aromatase and the samples classified in accordance with the number and intensity of stained cells. RESULTS: Aromatase was expressed positively in 32(71%) and negatively in 13(29%) of the cases in the IDC. The same results were obtained in the DCIS showing a perfect positive correlation. In the normal epithelium,aromatase was positive in 19(42.2%) and negative in 26 (57.8%) and a positive correlation, statistically significant was obtained when compared with IDC and DCIS(p<0.01). Concerning the normal stroma, positivity was only 7 (15.5%) showing no correlation with aromatase expression. Aromatase was positive in 36(80%) of the tumor stroma and this result was statistically significant as in the IDC and DCIS. Comparing results of aromatase expression with nuclear grade, histological grade, tumor size and age no difference was found. CONCLUSION: our results demonstrated high correlation between aromatase expression in IDC, DCIS, normal epithelium and tumor stroma showing a possible autocrine and paracrine mechanism of this enzyme in breast cancer.

Expressão da aromatase em carcinomas de mama ductais invasivo e in situ presentes na mesma mama / Aromatase expression in invasive and in situ ductal carcinoma present in the same breast

OBJETIVO: Avaliar expressão da enzima aromatase nos carcinomas de mama ductais invasivos (CDI), in situ (CDIS), no epitélio e estromas adjacentes. MÉTODOS: Foram avaliados 45 espécimes cirúrgicos provenientes de mastectomias e quadrantectomias com CDI e CDIS concomitantes de pacientes com estadios clínicos I e II. A análise da expressão da enzima aromatase foi realizada por meio de anticorpos policlonais antiaromatase e categorização das amostras de acordo com intensidade e número de células coradas. RESULTADOS: Nos 45 casos de CDI a expressão da aromatase foi positiva em 32 espécimes (71 por cento) e negativa em 13 (29 por cento). Nos casos de CDIS, a positividade foi idêntica à observada no CDI, mostrando correlação positiva. No epitélio normal constatou-se expressão positiva em 19 casos (42,2 por cento) e negativa nos outros 26 (57,8 por cento), mostrando correlação positiva estatisticamente (p<0,01), quando comparada com CDI e CDIS. Na análise do estroma normal a expressão da aromatase foi observada em apenas sete (15,5 por cento) dos 45 casos avaliados, não apresentando correlação com nenhuma variável analisada para expressão da aromatase. A presença da aromatase no estroma tumoral foi positiva em 36 casos (80 por cento) e negativa em 9 (20 por cento), mostrando correlação estatisticamente com a expressão no CDI (p<0,01) e no CDIS (p<0,01). Ao se comparar a expressão da aromatase no CDI, CDIS, epitélio normal e estroma tumoral com os graus nuclear e histológico, tamanho tumoral e idade da paciente, não foram encontradas diferenças estatisticamente significantes. CONCLUSÃO: Os resultados revelaram alta correlação entre expressão da aromatase no CDI, CDIS, epitélio normal e estroma tumoral, sugerindo possível mecanismo de ação autócrina e parácrina desta enzima na gênese do câncer de mama.

OBJECTIVE: to evaluate the expression of aromatase in simultaneously invasive ductal carcinoma (IDC) and ductal carcinoma in situ (DCIS). METHODS: forty-five surgical samples were obtained from mastectomy and quadrantectomy with simultaneous IDC and DCIS of stage I and II patients. Aromatase was evaluated using antibodies anti-aromatase and the samples classified in accordance with the number and intensity of stained cells. RESULTS: Aromatase was expressed positively in 32(71 percent) and negatively in 13(29 percent) of the cases in the IDC. The same results were obtained in the DCIS showing a perfect positive correlation. In the normal epithelium,aromatase was positive in 19(42.2 percent) and negative in 26 (57.8 percent) and a positive correlation, statistically significant was obtained when compared with IDC and DCIS(p<0.01). Concerning the normal stroma, positivity was only 7 (15.5 percent) showing no correlation with aromatase expression. Aromatase was positive in 36(80 percent) of the tumor stroma and this result was statistically significant as in the IDC and DCIS. Comparing results of aromatase expression with nuclear grade, histological grade, tumor size and age no difference was found. CONCLUSION: our results demonstrated high correlation between aromatase expression in IDC, DCIS, normal epithelium and tumor stroma showing a possible autocrine and paracrine mechanism of this enzyme in breast cancer.

Estudo urodinâmico da pressão de perda ao esforço, nas posições ortostática e sentada, em mulheres com incontinência urinária / Urodynamic evaluation of leak point pressure under stress, in orthostatic and seating position, in women with urinary incontinency

OBJETIVOS: comparar os resultados obtidos durante o estudo urodinâmico realizado em duas diferentes posições em relação às pressões de perda urinária sob esforço e discutir sua relevância clínica. MÉTODOS: sessenta e quatro pacientes com queixa de incontinência urinária de esforço (IUE) com idades variando entre 25 e 80 anos, atendidas no ambulatório de uroginecologia e cirurgia vaginal, no período de junho 2003 a setembro 2005 foram incluídas neste estudo. As pacientes foram inicialmente submetidas ao estudo urodinâmico de acordo com a técnica preconizada pela International Continency Society (ICS) na posição ortostática e logo depois foram avaliadas na posição sentada. RESULTADOS: diferença significante foi obtida após a avaliação das pressões de perda obtidas nas diferentes posições (99,8 ± 33,3 versus 102,9 ± 32,4; respectivamente, posição sentada e em pé, p < 0,05). Testes de regressão linear com análise de freqüência foram realizados com a finalidade de verificar a porcentagem de pacientes que ficaram dentro dos limites de confiança em relação às PP nas posições sentada e em pé. Uma taxa de 90,6 por cento de compatibilidade foi obtida nesses resultados. Quando três unidades foram somadas aos valores das pressões obtidas no estudo urodinâmico realizado na posição sentada, percebeu-se que 92,2 por cento ficaram inseridas neste intervalo. CONCLUSÕES: estes achados sugerem que o estudo urodinâmico pode ser realizado na posição sentada sem comprometimento diagnóstico e terapêutico proporcionando maior conforto e comodidade às pacientes.

PURPOSE: compare the outcomes verified during urodynamic investigation realized in two different positions related to urinary leak point pressure under stress and to discuss its clinical relevance. METHODS: sixty-four patients with stress urinary incontinency (SUI) aged 25-80 years old, attended, during June 2003 to September 2005 were included in this study. Patients were initially submitted to urodynamic investigation in accordance with International Continence Society (ICS) techniques in orthostatic position and just after were evaluated in seating position. RESULTS: statistical significance was obtained after evaluation of Vasalva leak point pressure (VLPP) obtained in two positions (99,8 ± 33,3 versus 102,9 ± 32,4; respectivamente, posição sentada e em pé, p<0,05). Linear regression test based on frequency analyses was applied with the purpose to verify the patient percentage allocated in confidence interval in terms of Valsalva leak point pressure in seating or orthostatic positions. A rate of 90.6 percent of compatibility was gotten in these results. When three unities were added to VLPP values after urodynamic investigation in seating position, it was noted that 92.2 percent of patients was included in this interval. CONCLUSIONS: these findings suggest that the urodynamic investigation can be realized in seating position without diagnostic a therapeutic impairment allowing higher comfort to the patients.

Fertilization and pregnancy outcome after intracytoplasmic injection with fresh or cryopreserved ejaculated spermatozoa.

OBJECTIVE: To assess fertilization, implantation, and pregnancy rates in patients undergoing ICSI using fresh and cryopreserved sperm from ejaculated semen samples. DESIGN: Retrospective study. SETTING: Academic and private medical centers. PATIENT(S): One hundred fifty-eight patients. INTERVENTION(S): Intracytoplasmic sperm injection. MAIN OUTCOME MEASURE(S): Fertilization, implantation, and pregnancy rates were evaluated; 61 cycles of ICSI were performed with cryopreserved and 79 cycles of ICSI were performed with fresh spermatozoa. Also, we divided the outcomes according to the semen characteristics, normozoospermia, oligozoospermia, asthenozoospermia, and oligoasthenozoospermia. RESULT(S): Overall, normal-fertilization rates were higher using fresh sperm (73.8%) compared with cryopreserved sperm (68.7%). Cycles performed in patients with normozoospermia or oligozoospermia had similar fertilization, implantation, and pregnancy rates using fresh or cryopreserved sperm. When asthenozoospermic and oligoasthenozoospermic semen samples were used, the normal-fertilization rate was higher with fresh sperm compared with cryopreserved sperm. However, implantation and pregnancy rates were similar in fresh and cryopreserved sperm samples from patients with asthenozoospermia or oligoasthenozoospermia. CONCLUSION(S): Semen with abnormalities in the motility may be more susceptible to sperm cryopreservation damage, resulting in lower fertilization rates. However, once the oocyte is fertilized, implantation and pregnancy rates are similar to those in patients with oligozoospermia and normozoospermia.

The impact of pronuclear orientation to select chromosomally normal embryos.

PURPOSE: To find an effective method to select embryos from ICSI with better chromosomal status when preimplantation-genetic-diagnosis (PGD) is not applied. METHODS: Several morpholological evaluations were done in same embryos. Embryos from ICSI were classified on day 1 according to pronuclear-nucleoli arrangement. On day 3, classification was done according to number, fragmentation, size and shape of cells. In some patients, embryos exhibiting good quality on day 3 (at least six regular blastomeres with cell fragmentation lower than 20%) were also submitted to PGD, irrespective to pronuclear-nucleoli morphology. RESULTS: Forty-two per cent of normally fertilized embryos showed pronuclear-nucleoli-good-morphology; from those, 86% were also classified as good quality on day 3. Good-quality embryos submitted to PGD have shown lower chromosomal abnormality rates when also classified as pronuclear-nucleoli-good-morphology. CONCLUSIONS: Pronuclear-nucleoli morphology seems to be correlated with PGD results. This criterion may prove useful for pre-select embryos with normal chromosomal package when PGD is not applied.

Etiology-specific outcomes of intracytoplasmic sperm injection in azoospermic patients.

OBJECTIVE: To assess fertilization, pregnancy, and miscarriage rates after intracytoplasmic sperm injection (ICSI) with epididymal or testicular spermatozoa from different types of azoospermia. DESIGN: Retrospective study. SETTING: Academic medical center and private fertility center. PATIENT(S): Two hundred twelve patients underwent 257 ICSI cycles. INTERVENTION(S): Cycles of ICSI were divided into four groups according to the etiology of azoospermia: A (nonobstruction), B (postvasectomy), C (congenital obstruction), and D (obstruction due to infection). Testicular sperm aspiration and percutaneous epididymal sperm aspiration were the sperm retrieval methods used for ICSI. MAIN OUTCOME MEASURE(S): Fertilization, pregnancy, and miscarriage rates. RESULT(S): Normal fertilization rates were higher in groups C (67.7%) and B (64.1%) compared with groups A (47.3%) and D (58.9%). Although lower pregnancy rates were seen in group A, no statistical differences were detected among groups. However, the miscarriage rate was higher in group A (45.6%) compared with groups B (25.25%), C (24%), and D (22.58%). CONCLUSION(S): Although no differences were detected in the pregnancy rates across groups, fertilization and implantation rates were higher in patients with congenital obstruction of the seminal path. The pregnancy rate was higher and the miscarriage rate lower when epididymal sperm was used compared with testicular sperm.

Sperm retrieval techniques in rats with suppressed spermatogenesis by experimental cryptorchidism.

OBJECTIVE: Our aim was to assess the suppression of spermatogenesis and sperm retrieval rate after testicular sperm extraction (TESE) or testicular sperm aspiration (TESA) in adult rats with surgically induced cryptorchidism. METHODS: Adult rats were submitted to TESE and TESA procedures after 15 days of induced cryptorchidism. After spermatozoa retrieval, the testicles were extracted, weighed and a morphological analysis by conventional light microscopy was done. The numbers of spermatozoa retrieved in both TESA and TESE were rated and compared. RESULTS: Histological analysis of the testicles revealed Sertoli cell-only syndrome in 60% of the testicles, and maturation arrest in the remaining cryptorchid testicles. Significant differences were seen in the number of spermatozoa retrieved (P < 0.05) between cryptorchidic and control rats. When sperm retrieval techniques were compared, no differences were detected in the number of spermatozoa obtained (P > 0.05). CONCLUSIONS: It seems that a 15 day period of cryptorchidism is enough to induce spermatogenesis disorders. No differences were detected in the number of spermatozoa retrieved in the right or left testicles, irrespective of the testicular pole. Furthermore, and even more importantly, no differences in the retrieval rate were seen between the two techniques.

Processamento seminal e utilização da técnica de ICSI em casais sorodiscordantes para o HIV-1 / Semen processing and ICSI technique in HIV-1 serodiscordant couples

Com os avanços na terapia antiretroviral para indivíduos infectados pelo HIV-1, aumentou a procura das técnicas de reprodução assistida por casais sorodiscordantes que almejam ter um filho biológico saudável.A associação de técnicas para processamento seminal em homens infectados pelo HIV-1 vêm sendo utilizadas com sucesso.Esse trabalho tem como objetivo relatar três ciclos de tratamento de reprodução assistida em dois casais sorodiscordantes, nos quais o homem é infectado pelo HIV-1.Em nosso estudo sugerimos uma modificação na técnica habitualmente utilizada para casais sorodiscordantes, que consiste na utilização de antiboticoterapia prévia com intuito de reduzir o número de linfócitos e macrófagos no ejaculado, seguida da associação de técnicas para processamento seminal, sperm wash, gradiente descontínuo de densidade e swim-up.A criopreservação e validação da amostra por PCR foram realizadas possibilitando maior confiabilidade para os procedimentos de reprodução assistida, e técnica de ICSI, que não depende do número final de espermatozóides móveis obtidos, além de contribuir para a redução no risco de transmissão do virus, parece ser o método de escolha nestes casos

Importância dos parâmetros seminais nos resultados de inseminaçäo intra-uterina / Importance of seminal parameters for intrauterine insemination outcomes

OBJETIVOS: analisar a relevância dos parâmetros seminais nos resultados dos ciclos de inseminaçäo intra-uterina (IIU) em pacientes com causa masculina de infertilidade e salientar o caráter prognóstico de cada parâmetro para o sucesso da técnica. MÉTODOS: duzentos e trinta e nove ciclos de IIU (155 casais) foram analisados durante período de 15 meses. Todos as pacientes foram submetidas à induçäo da ovulaçäo de acordo com protocolos do I Consenso Brasileiro de Induçäo de Ovulaçäo. A análise seminal foi realizada de acordo com os critérios da Organizaçäo Mundial da Saúde (OMS) para parâmetros como concentraçäo e motilidade e critério estrito de Kruger para avaliaçäo da morfologia. O preparo da amostra para IIU foi realizado por meio de gradientes descontínuos de densidade (ISolate®). Foram formados dois grupos de pacientes de acordo com o resultado de gestaçäo: grupo G - gestaçäo positiva e grupo NG - gestaçäo negativa. RESULTADOS: näo houve diferença significativa nos valores obtidos relacionados à concentraçäo total de espermatozóides/mL de sêmen, motilidade total, motilidade de formas progressivas antes e depois do processamento seminal. Quando a morfologia estrita de Kruger foi comparada entre os dois grupos, diferenças significativas foram encontradas (grupo G=10,6 por cento de formas normais; grupo NG=6,4 por cento de formas normais; p<0.05) quando o número de espermatozóides/mL inseminados foi superior a 15 milhöes/mL, a taxa de gravidez foi significantemente maior. CONCLUSÖES: tanto a morfologia estrita de Kruger como o número de espermatozóides inseminados parecem ser fatores prognósticos positivos para o estabelecimento da gestaçäo, devendo ser salientados na investigaçäo do homem infértil