Integrated molecular profiling of RAS, BRAF mutations, and mismatch repair status in advanced colorectal carcinoma: insights from gender and tumor laterality.

Background: Colorectal carcinoma (CRC) is one of the most frequently diagnosed forms of cancer worldwide. The RAS (KRAS, NRAS) and BRAF genes encode proteins that are important therapeutic targets for the treatment of CRC and, together with the mismatch repair (MMR) system, are closely related to patient prognosis and survival in advanced CRC. Here we evaluate the mutational profile and the frequency of mutations in the KRAS, NRAS and BRAF genes, along with the expression of MMR in advanced CRC, at a tertiary hospital in southern Brazil. Methods: A cross-sectional retrospective study was carried out, where molecular analysis of mutations in the KRAS, NRAS and BRAF genes was carried out, as well as immunohistochemistry for MMR proteins. Results: Next-generation sequencing (NGS) analysis of 310 tumors revealed that 202 patients (65.2%) had mutations. The KRAS gene (53.2%) was the most frequently mutated in our sample, with G12D being the most frequent, representing 30.5% of the mutations in this gene. The most frequent mutation found in BRAF was V600E (n=25; 89.3%) and differed significantly in women and in the right colon in patients with MMR deficiency. Among the 283 patients tested for MMR, the rate of loss of expression was 8.8% (25/283). Conclusions: Deficiency in the MMR system is associated with the presence of the BRAF V600E mutation, tumors located in the right colon, and the female sex. In our case series, more than 60% of patients had at least one mutation in KRAS, NRAS, or BRAF. The presence of mutations in these genes is closely related to CRC prognosis and helps define the best therapeutic approach in patients with metastatic CRC.

Gender affirming hormone therapy and transgender women fertility: Histologic predictors of germ cell presence.

Objective: Evaluate histological changes in testicular parameters after hormone treatment in transgender women. Methods: Cross-section study with patients who underwent gonadectomy at Hospital de Clínicas de Porto Alegre from 2011 to 2019. Hormone treatment type, route of administration, age at initiation and duration were recorded. Atrophy parameters were observed: testicular volume, tubular diameter, basal membrane length, presence of spermatogonia and spermatids (diploid and haploid spermatozoid precursors). Results: Eighty-six patients were included. Duration of hormone treatment is associated with testicular atrophy and spermatogenesis arrest. Other characteristics of hormone treatment such as age of initiation, route of administration and type of treatment were not associated with testicular histological changes. Testicular volume may predict spermatogenesis arrest. Basal membrane length and tubular diameter ratio is an interesting predictor of germ cell presence. Conclusion: Cross-sex hormone treatment affects testicular germ cell presence. Basal membrane length and tubular diameter ratio reduces inter variability of measurements and better exemplify how atrophic seminiferous tubules are. Fertility preservation should be addressed by healthcare providers in order to recognize gender affirming treatment impact on transgender health.

Gender affirming hormone therapy and transgender women fertility: Histologic predictors of germ cell presence

Abstract Objective: Evaluate histological changes in testicular parameters after hormone treatment in transgender women. Methods: Cross-section study with patients who underwent gonadectomy at Hospital de Clínicas de Porto Alegre from 2011 to 2019. Hormone treatment type, route of administration, age at initiation and duration were recorded. Atrophy parameters were observed: testicular volume, tubular diameter, basal membrane length, presence of spermatogonia and spermatids (diploid and haploid spermatozoid precursors). Results: Eighty-six patients were included. Duration of hormone treatment is associated with testicular atrophy and spermatogenesis arrest. Other characteristics of hormone treatment such as age of initiation, route of administration and type of treatment were not associated with testicular histological changes. Testicular volume may predict spermatogenesis arrest. Basal membrane length and tubular diameter ratio is an interesting predictor of germ cell presence. Conclusion: Cross-sex hormone treatment affects testicular germ cell presence. Basal membrane length and tubular diameter ratio reduces inter variability of measurements and better exemplify how atrophic seminiferous tubules are. Fertility preservation should be addressed by healthcare providers in order to recognize gender affirming treatment impact on transgender health.

Ovarian Tissue Culture to Preserve Fertility in Transgender Male Patients after Hormonal Treatment / Cultura de tecido ovariano para preservação da fertilidade em pacientes transexuais masculinos após tratamento hormonal

Abstract Objective To evaluate the reproductive and histological characteristics of fresh cultured ovarian tissue from transgender male patients. Methods An in vitro pilot study in which samples were collected during sex reassignment surgery for transgender male patients. The ovarian cortex was cut into fragments of 2 mm, 3mm, and 4 mm, and placed in a 96-well plate suitable for cultivation at days 0, 2, 4, 6, and 8, when the histology was analyzed. Results Stromal hyperplasia was observed in all samples, and it was not associated with the obtainment of primordial or primary follicles. Peripheral reduction in cell count was also a recurrent finding. Primordial and primary follicles were identified with a heterogeneous pattern in fragments from the same patient and from different patients, and follicles in more advanced stages of development (secondary and antral) were not found. There was an association between the diameter of the ovarian fragments and the identification of primary follicles (p=0.036). The number of days in culture was associated with histological signs of tissue damaging in the fragments (p=0.002). The total number of follicles identified in the samples with 2mm in diameter was significantly lower than in those that measured 4mm in diameter (p=0.031). Conclusion A diameter of 4mm is suitable for ovarian tissue culture with the benefit of ease of handling. Even after prolonged exposure to testosterone, the ovarian fragments presented primordial and primary follicles, maintaining viability throughout the days they were exposed to the culture. Freezing the ovarian cortex of transgender patients who will undergo surgery for gender reassignment would be an interesting option, in the future, for the preservation of fertility.

Resumo Objetivo Avaliar as características reprodutivas e histológicas de tecido ovariano cultivado a fresco de pacientes transexuais masculinos. Métodos Estudo experimental in vitro e piloto, no qual amostras foram coletadas durante a cirurgia de redesignação de sexo para pacientes transexuais masculinos. O córtex ovariano foi cortado em fragmentos de 2mm, 3mm, e 4mm, e colocado em placa de 96 poços própria para cultivo nos dias 0, 2, 4, 6 e 8, quando a histologia foi analisada. Resultados Hiperplasia estromal foi observada em todas as amostras, e não esteve associada à obtenção de folículos primordiais ou primários. A redução periférica no número de células também foi um achado recorrente. Folículos primordiais e primários foramidentificados com padrão heterogêneo emfragmentos domesmo paciente e em fragmentos de pacientes diferentes, não sendo encontrados folículos em estágios mais avançados de desenvolvimento (secundários e antrais). Houve associação entre o diâmetro dos fragmentos ovarianos e a identificação dos folículos primários (p=0,036). O número de dias de cultura esteve associado a sinais histológicos de lesão tecidual nos fragmentos (p=0,002). O número total de folículos identificados nas amostras de 2mm de diâmetro foi significativamente menor do que nas de 4mm de diâmetro (p=0,031). Conclusão O diâmetro de 4mm parece ser mais adequado para a cultura de tecido ovariano com a vantagem de fácil manejo. Mesmo após exposição prolongada à testosterona, os fragmentos ovarianos apresentavam folículos primordiais e primários, e manteve a viabilidade ao longo dos dias de exposição à cultura. No futuro, o congelamento da cortical do ovário de pacientes transgêneros que se submeterão à cirurgia de redesignação sexual poderia ser uma opção interessante para a preservação da fertilidade.

Ovarian Tissue Culture to Preserve Fertility in Transgender Male Patients after Hormonal Treatment.

OBJECTIVE: To evaluate the reproductive and histological characteristics of fresh cultured ovarian tissue from transgender male patients. METHODS: An in vitro pilot study in which samples were collected during sex reassignment surgery for transgender male patients. The ovarian cortex was cut into fragments of 2 mm, 3 mm, and 4 mm, and placed in a 96-well plate suitable for cultivation at days 0, 2, 4, 6, and 8, when the histology was analyzed. RESULTS: Stromal hyperplasia was observed in all samples, and it was not associated with the obtainment of primordial or primary follicles. Peripheral reduction in cell count was also a recurrent finding. Primordial and primary follicles were identified with a heterogeneous pattern in fragments from the same patient and from different patients, and follicles in more advanced stages of development (secondary and antral) were not found. There was an association between the diameter of the ovarian fragments and the identification of primary follicles (p = 0.036). The number of days in culture was associated with histological signs of tissue damaging in the fragments (p = 0.002). The total number of follicles identified in the samples with 2 mm in diameter was significantly lower than in those that measured 4 mm in diameter (p = 0.031). CONCLUSION: A diameter of 4 mm is suitable for ovarian tissue culture with the benefit of ease of handling. Even after prolonged exposure to testosterone, the ovarian fragments presented primordial and primary follicles, maintaining viability throughout the days they were exposed to the culture. Freezing the ovarian cortex of transgender patients who will undergo surgery for gender reassignment would be an interesting option, in the future, for the preservation of fertility.

OBJETIVO: Avaliar as características reprodutivas e histológicas de tecido ovariano cultivado a fresco de pacientes transexuais masculinos. MéTODOS: Estudo experimental in vitro e piloto, no qual amostras foram coletadas durante a cirurgia de redesignação de sexo para pacientes transexuais masculinos. O córtex ovariano foi cortado em fragmentos de 2 mm, 3 mm, e 4 mm, e colocado em placa de 96 poços própria para cultivo nos dias 0, 2, 4, 6 e 8, quando a histologia foi analisada. RESULTADOS: Hiperplasia estromal foi observada em todas as amostras, e não esteve associada à obtenção de folículos primordiais ou primários. A redução periférica no número de células também foi um achado recorrente. Folículos primordiais e primários foram identificados com padrão heterogêneo em fragmentos do mesmo paciente e em fragmentos de pacientes diferentes, não sendo encontrados folículos em estágios mais avançados de desenvolvimento (secundários e antrais). Houve associação entre o diâmetro dos fragmentos ovarianos e a identificação dos folículos primários (p = 0,036). O número de dias de cultura esteve associado a sinais histológicos de lesão tecidual nos fragmentos (p = 0,002). O número total de folículos identificados nas amostras de 2 mm de diâmetro foi significativamente menor do que nas de 4 mm de diâmetro (p = 0,031). CONCLUSãO: O diâmetro de 4 mm parece ser mais adequado para a cultura de tecido ovariano com a vantagem de fácil manejo. Mesmo após exposição prolongada à testosterona, os fragmentos ovarianos apresentavam folículos primordiais e primários, e manteve a viabilidade ao longo dos dias de exposição à cultura. No futuro, o congelamento da cortical do ovário de pacientes transgêneros que se submeterão à cirurgia de redesignação sexual poderia ser uma opção interessante para a preservação da fertilidade.

Reconstruction of parietal bone defects with adiposederived mesenchymal stem cells. Experimental study.

PURPOSE: This study assessed the regeneration potential of mesenchymal stem cells (MSC) from adipose tissue associated with platelet-rich plasma (PRP) in bone regeneration. METHODS: Thirty Wistar rats (Rattus norvegicus albinos) were divided into five groups (according to the grafting material and time to euthanasia): (1) autograft - 14 days (control), (2) autograft - 28 days (control), (3) MSC + PRP - 14 days, (4) MSC + PRP + papaverine - 14 days and (5) MSC + PRP + papaverine - 28 days. After euthanasia, the graft was removed and histological slides were prepared. They were assessed by a blinded pathologist using a previously published histological scale as parameter. RESULTS: There was some degree of neoformed bone trabeculae (NBT) in 93.3% of the samples, as well as osteoblastic activity (OA). The autograft groups (14 and 28 days) had higher levels in the formation of bone trabeculae. Nonparametric data were analyzed using the Wilcoxon-Mann-Whitney test and proved not to be statistically significant at p < 0.05. CONCLUSIONS: Experimental parietal bone reconstruction, combining MSC, PRP and papaverine presented regeneration in all groups with no significant difference among them.

Avaliação do perfil de mulheres atendidas em centros de referência em saúde de Porto Alegre/RS e relação de alterações citológicas detectadas no exame citopatológico e a presença do HPV / Profile evaluation of women that attended reference health centers in Porto Alegre/RS and the relationship between cytological alterations detected in cytopathological examination and presence of HPV / Evaluación del perfil de las mujeres atendidas en los centros de referencia de salud de Porto Alegre / RS y la relación de las alteraciones citológicas detectadas en el examen citopatológico con la presencia de VPH

Justificativas e Objetivos: Câncer de colo de útero é considerado um problema de saúde pública mundial. Seu diagnóstico é realizado através do exame citopatológico (EC) e seu desenvolvimento relacionado à infecção pelo papilomavírus humano (HPV). Este estudo objetiva avaliar o perfil de mulheres atendidas em centros de referência em saúde de Porto Alegre, Rio Grande do Sul, assim como a relação de alterações observadas ao EC com presença do HPV. Métodos: Estudo transversal realizado em mulheres atendidas em unidades básicas de saúde e um ambulatório de referência de hospital público terciário, no período de julho de 2014 a janeiro de 2017. Coletaram-se amostras representativas da endo/ectocérvice para realização do EC e investigadas quanto à presença molecular do HPV. Resultados: Foram analisadas 169 mulheres com idade média entre 31 e 40 anos, das quais 125 (74%) informaram que a sexarca ocorreu na faixa de 15-20 anos e 37,9% relatou ter tido de três a cinco parceiros sexuais. Em relação ao EC, 71 (42%) apresentaram resultado negativo para lesão intraepitelial ou malignidade e 98 (58%) alguma anormalidade de células escamosas: 20 (11,8%) atipias; 22 (13%) lesão intraepitelial escamosa de baixo grau e 56 (32,6%) lesão intraepitelial de alto grau (HSIL). Cinquenta (29,6%) apresentaram positividade para HPV, destas 56,4% foram diagnosticadas com HSIL (p<0,01). Conclusão: Os resultados revelam alta frequência de HPV em amostras com alterações citopatológicas, em mulheres jovens e com grau de exposição ao HPV, reforçando a importância do papel da sua identificação precoce na investigação da carcinogênese cervical.(AU)

Background and Objectives: Cervical cancer is considered a worldwide public health problem. Its diagnosis is made through cytopathological examination and its development related to human papillomavirus (HPV) infection. This study aims to evaluate the profile of women treated at reference health centers in Porto Alegre, Rio Grande do Sul, as well as the relation of changes observed to cytopathological examination with the presence of HPV. Methods: This is a cross-sectional study carried out in women treated at basic health units and a referral clinic of a public tertiary hospital, from July 2014 to January 2017. Representative samples of the endo/ectocervix were collected to perform the cytopathological examination and investigated for the molecular presence of HPV. Results: 169 women with mean age between 31 and 40 years were analyzed, of whom 125 (74%) reported that the onset of sexual activity occurred in the 15-20 years age group, and 37.9% reported having had three to five sexual partners. In relation to cytopathological examination, 71 (42%) had a negative result for intraepithelial lesion or malignancy and 98 (58%) some squamous cell abnormality: 20 (11.8%) atypical; 22 (13%) low-grade squamous intraepithelial lesion and 56 (32.6%) high-grade intraepithelial lesion (HSIL). Fifty (29.6%) were positive for HPV, of which 56.4% were diagnosed with HSIL (p < 0.01). Conclusion: The results reveal a high frequency of HPV in samples with cytopathological changes, in young women and with a degree of exposure to HPV, reinforcing the importance of the role of its early identification in the investigation of cervical carcinogenesis.(AU)

Justificación y objetivos: El cáncer de cuello uterino se considera un problema de salud pública en todo el mundo. Su diagnóstico se realiza mediante el examen citopatológico (EC), y su desarrollo está relacionado con la infección por el virus del papiloma humano (VPH). Este estudio objetivó evaluar el perfil de mujeres atendidas en los centros de referencia en salud de Porto Alegre, Rio Grande do Sul (Brasil), así como la relación de las alteraciones observadas en el EC con la presencia del VPH. Métodos: Estudio transversal realizado en las mujeres atendidas por unidades de atención primaria y por una clínica ambulatoria de referencia del hospital público terciario en la ciudad de Porto Alegre, en el período de julio de 2014 a enero de 2017. Se recolectaron muestras representativas de endo/ectocérvice para realizar la CE, las cuales se clasificaron según el sistema Bethesda y se investigaron la presencia molecular del VPH. Resultados: Analizamos 169 mujeres con promedio de edad entre 31 y 40 años, de las cuales 125 (74%) informaron que el sexarche ocurrió en el rango de 15-20 años. La mayoría (37,9%) informó haber tenido de 3 a 5 parejas sexuales; y el 37,3% estaban usando anticonceptivos orales. Con respecto a la EC, 71 (42%) se clasificaron como negativos para lesión intraepitelial o malignidad; y el 98 (58%) tenían alguna anormalidad de células escamosas: 20 (11,8%) de atipias; 22 (13,0%) lesión intraepitelial escamosa de bajo grado y 56 (32,6%) lesión intraepitelial de alto grado (HSIL). La frecuencia de positividad del VPH encontrada fue de 50 (29,6%), de estas un 56,4% fueron diagnosticadas con HSIL (p<0,01). Conclusiones: Estos resultados revelan una alta frecuencia de VPH en muestras con alteraciones citopatológicas presentes en mujeres jóvenes con cierto grado de exposición al VPH, lo que refuerza la importancia de identificarse tempranamente en el análisis de la carcinogénesis cervical.(AU)

Stanniocalcin-1 protein expression profile and mechanisms in proliferation and cell death pathways in prostate cancer.

Prostate cancer (PCa) is one of the most prevalent male tumours. Stanniocalcin-1 (STC1) is a glycoprotein and, although the role of STC1 in human cancer is poorly understood, it is suggested to be involved in the development and progression of different neoplasms. This study investigated the protein expression profile of STC1 in PCa and benign prostatic hyperplasia (BPH) samples and STC1 signalling during cell proliferation and cell death in vitro using cell lines. We found higher levels of STC1 in PCa when compared to BPH tissue and that STC1 inhibited forskolin stimulation of cAMP in PC-3 cells. A monoclonal antibody against STC1 was effective in reducing cell proliferation, in promoting cell cycle arrest, and in increasing apoptosis in the same cells. Since STC1 acts as a regulator of prostatic tissue signalling, we suggest that this protein is a novel candidate biomarker for prostate tumour clinical progression and a potential therapeutic target.

Reconstruction of parietal bone defects with adiposederived mesenchymal stem cells. Experimental study

Abstract Purpose: This study assessed the regeneration potential of mesenchymal stem cells (MSC) from adipose tissue associated with platelet-rich plasma (PRP) in bone regeneration. Methods: Thirty Wistar rats (Rattus norvegicus albinos) were divided into five groups (according to the grafting material and time to euthanasia): (1) autograft - 14 days (control), (2) autograft - 28 days (control), (3) MSC + PRP - 14 days, (4) MSC + PRP + papaverine - 14 days and (5) MSC + PRP + papaverine - 28 days. After euthanasia, the graft was removed and histological slides were prepared. They were assessed by a blinded pathologist using a previously published histological scale as parameter. Results: There was some degree of neoformed bone trabeculae (NBT) in 93.3% of the samples, as well as osteoblastic activity (OA). The autograft groups (14 and 28 days) had higher levels in the formation of bone trabeculae. Nonparametric data were analyzed using the Wilcoxon-Mann-Whitney test and proved not to be statistically significant at p < 0.05. Conclusions: Experimental parietal bone reconstruction, combining MSC, PRP and papaverine presented regeneration in all groups with no significant difference among them.

Which mode and potency of electrocoagulation yields the Smallest Unobstructed Area of the Fallopian Tubes? / Qual modo e potência produzem a menor área de nãoobstrução nas tubas de Falópio?

Abstract Objective To determine which mode and potency of electrocoagulation, using a modern electrosurgical generator, yields the smallest unobstructed area of the Fallopian tubes. Methods In an experimental study, tubes from 48 hysterectomies or tubal ligation were evaluated. Tubes were randomly allocated to one of the following groups: group A) 25 W x 5 seconds (n = 17); group B) 30 W x 5 seconds (n = 17); group C) 35 W x 5 seconds (n = 18), group D) 40 W x 5 seconds (n = 20); group E) 40 W x 5 seconds with visual inspection (blanch, swells, collapse) (n = 16); group F) 50 W x 5 seconds (n = 8). Bipolar electrocoagulation was performed in groups A to E, and monopolar electrocoagulation was performed in group F. Coagulation mode was used in all groups. Digital photomicrography of the transversal histological sections of the isthmic segment of the Fallopian tube were taken, and themedian percentage of unobstructed luminal area (mm2) was measured with ImageJ software (ImageJ, National Institutes of Health, Bethesda, MD, USA). The Kruskal-Wallis test or analysis of variance (ANOVA) was used for statistical analysis. Results Ninety-six Fallopian tube sections were analyzed. The smallest median occluded area (%; range) of the Fallopian tube was obtained in the group with 40 W with visual inspection (8.3%; 0.9-40%), followed by the groups 25 W (9.1%; 0-35.9%), 40 W (14.2; 0.9-43.2%), 30 W (14.2; 0.9-49.7%), 35 W (15.1; 3-46.4%) and 50 W (38.2; 3.1-51%). No statistically significant difference was found among groups (p = 0.09, Kruskal-Wallis test). Conclusion The smallest unobstructed area was obtained with power setting at 40 W with visual inspection using a modern electrosurgical generator. However, no statistically significant difference in the unobstructed area was observed among the groups using these different modes and potencies.

Resumo Objetivo Determinar em qual modo e potência, usando unidades geradoras modernas de eletrocoagulação, produz a menor área de não-obstrução das tubas de Falópio. Métodos Num estudo experimental, tubas uterinas derivadas de 48 histerectomias ou ligadura tubária foram avaliadas. As tubas foram alocadas aleatoriamente para um dos seguintes grupos: grupo A) 25 W x 5 segundos (n = 17); grupo B) 30 W x 5 segundos (n = 17); grupo C) 35 Wx 5 segundos (n = 18), grupo D) 40 W, 5 segundos (n = 20); grupo E) 40 W x 5 segundos inspeção visual (branqueia, incha e colapsa) (n = 16); grupo F) 50 Wx 5 segundos (n = 8). A eletrocoagulação bipolar foi usada nos grupos de A a E, e a eletrocoagulação monopolar, no grupo F. O modo de coagulação foi utilizado em todos os grupos. Cortes histológicos transversais do segmento ístmico das tubas de Falópio foram corados e fotografados digitalmente, e a percentagem da área luminal (mm2) não-obstruída foi medida com o software ImageJ (ImageJ, National Institutes of Health, Bethesda, MD, USA). O teste de Kruskal-Wallis ou ANOVA foram usados para a análise estatística. Resultados Noventa e seis cortes histológicos de tubas de Falópio foram analisados. A mediana da menor área não-obstruída (%; amplitude) da tuba de Falópio foi obtida no grupo 40 W com inspeção visual (8,3%; 0,9-40%), seguido do grupo 25 W (9,1%; 0- 35,9%), 40W(14,2; 0,9-43,2%), 30 W(14.2; 0,9-49,7%), 35 W(15,1; 3-46,4%) e 50 W (38,2; 3.1-51%). Não houve diferença significativa entre os grupos (p = 0,09, teste de Kruskal-Wallis). Conclusão A menor área não-obstruída foi obtida com a potência de 40 W com inspeção visual usando um gerador moderno de eletrocirurgia. Contudo, nenhuma diferença significativa na área não-obstruída foi observada entre os grupos usando esses modos e potências.

Which mode and potency of electrocoagulation yields the Smallest Unobstructed Area of the Fallopian Tubes?

OBJECTIVE: To determine which mode and potency of electrocoagulation, using a modern electrosurgical generator, yields the smallest unobstructed area of the Fallopian tubes. METHODS: In an experimental study, tubes from 48 hysterectomies or tubal ligation were evaluated. Tubes were randomly allocated to one of the following groups: group A) 25 W x 5 seconds (n = 17); group B) 30 W x 5 seconds (n = 17); group C) 35 W x 5 seconds (n = 18), group D) 40 W x 5 seconds (n = 20); group E) 40 W x 5 seconds with visual inspection (blanch, swells, collapse) (n = 16); group F) 50 W x 5 seconds (n = 8). Bipolar electrocoagulation was performed in groups A to E, and monopolar electrocoagulation was performed in group F. Coagulation mode was used in all groups. Digital photomicrography of the transversal histological sections of the isthmic segment of the Fallopian tube were taken, and the median percentage of unobstructed luminal area (mm2) was measured with ImageJ software (ImageJ, National Institutes of Health, Bethesda, MD, USA). The Kruskal-Wallis test or analysis of variance (ANOVA) was used for statistical analysis. RESULTS: Ninety-six Fallopian tube sections were analyzed. The smallest median occluded area (%; range) of the Fallopian tube was obtained in the group with 40 W with visual inspection (8.3%; 0.9-40%), followed by the groups 25 W (9.1%; 0-35.9%), 40 W (14.2; 0.9-43.2%), 30 W (14.2; 0.9-49.7%), 35 W (15.1; 3-46.4%) and 50 W (38.2; 3.1-51%). No statistically significant difference was found among groups (p = 0.09, Kruskal-Wallis test). CONCLUSION: The smallest unobstructed area was obtained with power setting at 40 W with visual inspection using a modern electrosurgical generator. However, no statistically significant difference in the unobstructed area was observed among the groups using these different modes and potencies.

OBJETIVO: Determinar em qual modo e potência, usando unidades geradoras modernas de eletrocoagulação, produz a menor área de não-obstrução das tubas de Falópio. MéTODOS: Num estudo experimental, tubas uterinas derivadas de 48 histerectomias ou ligadura tubária foram avaliadas. As tubas foram alocadas aleatoriamente para um dos seguintes grupos: grupo A) 25 W x 5 segundos (n = 17); grupo B) 30 W x 5 segundos (n = 17); grupo C) 35 W x 5 segundos (n = 18), grupo D) 40 W, 5 segundos (n = 20); grupo E) 40 W x 5 segundos inspeção visual (branqueia, incha e colapsa) (n = 16); grupo F) 50 W x 5 segundos (n = 8). A eletrocoagulação bipolar foi usada nos grupos de A a E, e a eletrocoagulação monopolar, no grupo F. O modo de coagulação foi utilizado em todos os grupos. Cortes histológicos transversais do segmento ístmico das tubas de Falópio foram corados e fotografados digitalmente, e a percentagem da área luminal (mm2) não-obstruída foi medida com o software ImageJ (ImageJ, National Institutes of Health, Bethesda, MD, USA). O teste de Kruskal-Wallis ou ANOVA foram usados para a análise estatística. RESULTADOS: Noventa e seis cortes histológicos de tubas de Falópio foram analisados. A mediana da menor área não-obstruída (%; amplitude) da tuba de Falópio foi obtida no grupo 40 W com inspeção visual (8,3%; 0,9­40%), seguido do grupo 25 W (9,1%; 0­35,9%), 40W (14,2; 0,9­43,2%), 30 W (14.2; 0,9­49,7%), 35 W (15,1; 3­46,4%) e 50 W (38,2; 3.1­51%). Não houve diferença significativa entre os grupos (p = 0,09, teste de Kruskal-Wallis). CONCLUSãO: A menor área não-obstruída foi obtida com a potência de 40 W com inspeção visual usando um gerador moderno de eletrocirurgia. Contudo, nenhuma diferença significativa na área não-obstruída foi observada entre os grupos usando esses modos e potências.

Carcinogenesis of Vulvar Lesions: Morphology and Immunohistochemistry Evaluation.

OBJECTIVES: The aim of the study was to assess the 2 pathways of vulvar carcinogenesis and correlate immunohistochemical expression of p53 with histopathological findings. MATERIALS AND METHODS: This cross-sectional study included 76 cases. Patients were classified according to the 2004 International Society for the Study of Vulvovaginal Disease Terminology, followed by a review of clinical records and immunohistochemical staining for p53. RESULTS: Fifteen cases were in the human papillomavirus (HPV)-associated pathway (12 cases of usual vulvar intraepithelial neoplasia [VIN] and 3 of warty squamous cell carcinoma [SCC]), and 13 cases were in the HPV-independent pathway (5 cases of differentiated VIN and 8 of keratinizing SCC). Significant differences in p53 expression were observed between the 2 pathways of carcinogenesis: in the lesions related to the HPV-independent pathway, the percentage of p53-positive cells was greater (>25%, p < .001), and the staining pattern was basal (extending into the middle layer) in differentiated VIN and diffuse or infiltrative in warty SCC (p < 0.001). In the lesions HPV-associated pathway, p53 staining was less extensive (≤10% of cells, p < 0.001) and followed basal pattern in usual VIN, whereas warty SCCs were negative for p53 (p < 0.001). CONCLUSIONS: Unique patterns of histological appearance and p53 expression can separate vulvar lesions into 2 distinct pathways of carcinogenesis. We propose that p53 immunohistochemistry may be performed simultaneously with histopathological examination in all cases of VIN and vulvar SCC, because it would aid in definition of the pathway of carcinogenesis and thus enable better clinical follow-up of patients with these conditions.

p14 expression differences in ovarian benign, borderline and malignant epithelial tumors.

BACKGROUND: Abnormalities in tumor suppressors p14, p16 and p53 are reported in several human cancers. In ovarian epithelial carcinogenesis, p16 and p53 show higher immunohistochemical staining frequencies in malignant tumors and are associated with poor prognoses. p14 was only analyzed in carcinomas, with conflicting results. There are no reports on its expression in benign and borderline tumors. This study aims to determine p14, p16 and p53 expression frequencies in ovarian benign, borderline and malignant tumors and their associations with clinical parameters. METHODS: A cross-sectional study utilizing immunohistochemistry was performed on paraffin-embedded ovarian epithelial tumor samples. Clinical data were collected from medical records. Fisher's exact test and the Bonferroni correction were performed for frequency associations. Survival comparisons utilized Kaplan-Meier and log rank testing. Associations were considered significant when p < 0.05. RESULTS: p14 absent expression was associated with malignant tumors (60 % positive) (p = 0.000), while 93 % and 94 % of benign and borderline tumors, respectively, were positive. p16 was positive in 94.6 % of carcinomas, 75 % of borderline and 45.7 % of benign tumors (p = 0.000). p53 negative staining was associated with benign tumors (2.9 % positive) (p = 0.016) but no difference was observed between borderline (16.7 %) and malignant tumors (29.7 %) (p = 0.560). No associations were found between expression rates, disease-free survival times or clinical variables. Carcinoma subtypes showed no difference in expression. CONCLUSIONS: This is the first description of p14 expression in benign and borderline tumors. It remains stable in benign and borderline tumors, while carcinomas show a significant absence of staining. This may indicate that p14 abnormalities occur later in carcinogenesis. p16 and p53 frequencies increase from benign to borderline and malignant tumors, similarly to previous reports, possibly reflecting the accumulation of inactive mutant protein. The small sample size may have prevented statistically significant survival analyses and clinical correlations. Future studies should investigate genetic abnormalities in p14 coding sequences and include all types of ovarian epithelial tumors. Bigger sample sizes may be needed for significant associations.

Analysis of the effects of topical renal hypothermia on lung tissue after kidney ischemia and reperfusion in rats.

PURPOSE: To evaluate whether topical renal hypothermia (TRH) at different levels of temperature has protective effects on lung tissue after renal I/R, through an analysis of organ histology and inflammatory markers in lung tissue. METHODS: Twenty-eight male Wistar rats were randomly allocated across four groups and subjected to renal ischemia at different levels of topical renal temperature: normothermia (no cooling, 37°C), mild hypothermia (26°C), moderate hypothermia (15°C), and deep hypothermia (4°C). To induce I/R, the vessels supplying the left kidney of each animal were clamped for 40 minutes, followed by reperfusion. After four hours, another procedure was performed to harvest the tissues of interest. TNF-α, IL-1ß and myeloperoxidase activity were measured in lung tissue. Histological analysis was performed in hematoxylin and eosin-stained lung specimens. RESULTS: Induction of renal I/R under deep topical hypothermia resulted in a significant decrease in lung concentrations of TNF-α compared with normothermic I/R (p<0.05). A trend toward significant correlation was found between lung IL-1ß concentration and intensity of hypothermia (Spearman r=-0.37; p=0.055). No difference was found in myeloperoxidase activity or histologic injury between groups. CONCLUSION: Topical renal hypothermia reduces activation of the inflammatory cascade in the lung parenchyma. However, tissue-protective effects were not observed.

Analysis of the effects of topical renal hypothermia on lung tissue after kidney ischemia and reperfusion in rats

PURPOSE: To evaluate whether topical renal hypothermia (TRH) at different levels of temperature has protective effects on lung tissue after renal I/R, through an analysis of organ histology and inflammatory markers in lung tissue. METHODS: Twenty-eight male Wistar rats were randomly allocated across four groups and subjected to renal ischemia at different levels of topical renal temperature: normothermia (no cooling, 37°C), mild hypothermia (26°C), moderate hypothermia (15°C), and deep hypothermia (4°C). To induce I/R, the vessels supplying the left kidney of each animal were clamped for 40 minutes, followed by reperfusion. After four hours, another procedure was performed to harvest the tissues of interest. TNF-α, IL-1β and myeloperoxidase activity were measured in lung tissue. Histological analysis was performed in hematoxylin and eosin-stained lung specimens. RESULTS: Induction of renal I/R under deep topical hypothermia resulted in a significant decrease in lung concentrations of TNF-α compared with normothermic I/R (p<0.05). A trend toward significant correlation was found between lung IL-1β concentration and intensity of hypothermia (Spearman r=−0.37; p=0.055). No difference was found in myeloperoxidase activity or histologic injury between groups. CONCLUSION: Topical renal hypothermia reduces activation of the inflammatory cascade in the lung parenchyma. However, tissue-protective effects were not observed. .

Adenocarcinoma de endométrio: epidemiologia, tratamento e sobrevida de pacientes atendidas no Hospital de Clínicas de Porto Alegre / Endometrial adenocarcinoma: epidemiology, treatment and survival of patients treated at Hospital de Clínicas de Porto Alegre

Neste estudo, descreve-se o perfil clínico das pacientes e as características histopatológicas dos carcinomas de endométrio tratados no setor de Oncologia Genital do Hospital de Clínicas de Porto Alegre (HCPA), assim como as formas de tratamento, fatores prognósticos e sobrevida. Métodos: Estudo de coorte histórica incluindo todas as pacientes submetidas a tratamento cirúrgico primário entre 1996 e 2012. Após revisão de prontuários médicos, foram analisadas as variáveis idade, status hormonal, tipo histológico e grau tumoral, invasão miometrial, estadiamento cirúrgico, cirurgia realizada, tratamento complementar e sobrevida. Resultados: Cento e sessenta e quatro pacientes foram incluídas no estudo, com idade média de 64,2 anos (31-95 anos), sendo quase 90% delas pós-menopáusicas. O tempo de seguimento variou de 4 dias a 14,6 anos. O tipo histológico endometrioide foi o mais encontrado (78% dos casos). A histerectomia com salpingo-ooforectomia bilateral com linfadenectomia pélvica foi a cirurgia mais realizada (77,5%). Tratamento complementar foi realizado em 57,9% das pacientes, sendo a radioterapia o tratamento de escolha em 87,4% deles. Ocorreram 36 óbitos (22%) durante o seguimento, com uma sobrevida média global de 125 meses. Em análise bivariada, idade ≥ 65 anos, tipo histológico não endometrioide, tumores pouco diferenciados (G3), invasão miometrial ≥ 50% e metástase linfonodal relacionaram-se significativamente a um menor tempo de sobrevida. Em análise multivariada, a histologia não endometrioide, estádio III, estádio IV e a presença de comprometimento linfonodal foram significativamente associados ao óbito. Conclusão: Os resultados encontrados são compatíveis com a literatura existente e vêm em acréscimo à escassa estatística nacional...

This study describes the clinical profile and the hystopathologic characteristics of endometrial carcinomas from patients treated at the Gynecologic Oncology department of Hospital de Clínicas de Porto Alegre (HCPA), as well as the forms of treatment, prognostic factors, and survival. Methods: Historic cohort study including all patients subjected to primary surgical treatment between 1996 and 2012. After review of the medical records, the variablesage, hormonal status, tumor histologic type and grade, myometrial invasion, surgical staging, performed surgery, complementary treatment, and survival were analyzed. Results: One hundred sixty four patients were included, with a mean age of 64.2 years (31-95 years), of which almost 90% were postmenopausal women. Follow-up time ranged from 4 days to 14.6 years. Endometrioid adenocarcinoma was the most frequently histological type (78% of cases). Hysterectomy with bilateral salpingooophorectomy plus pelvic linfadenectomy was the most frequently performed surgery (77.5%). Adjuvant treatment was held in 57.9% of the patients, with radiotherapy being the treatment of choice in 87.4%. Thirty-six deaths (22%) occurred during followup, with a mean overall survival of 125 months. In the bivariate analysis, age ≥ 65 years, non-endometrioid histology, poorly differentiated tumors (G3), myometrial invasion ≥ 50%, and lymph node metastasis were correlated to lower survival. In the multivariate analysis, non-endometrioid histology, stage III, stage IV and lymph node metastasis were significantly associated with death. Conclusion: The results found are compatible with the existing literature and contribute to the scarce existing national statistics...

Experimental histopathological study on retinal and renal cellular response to intravitreous antiangiogenic drugs.

AIM: To evaluate histopathological retinal and renal response after one single dose of intravitreous injection of antiangiogenic drugs ranibizumab and bevacizumab in rats. METHODS: Experimental study in 60d of life adults Wistar rats. Ten animals were included. Group 1 included 5 animals that were injected with 1 µL ranibizumab 1.25 mg in the right eye and with 1 µL of balanced salt solution (BSS) in the left eye, as control; Group 2 included 5 animals that were injected with 1 µL of bevacizumab in the right eye and with 1 µL of BSS in the fellow eye. All injections were performed with Hamilton syringes. After 15d of the interventions, all animals were sacrificed in CO2 chamber. Both eyes were enucleated and one kidney was removed, fixed and embedded in paraffin for histopathological analysis by optic microscopy. For statistical purposes the initial expected abnormal histopathological responses were defined as 0%. RESULTS: Atypical histopathological retinal response was detected in 2 eyes injected with ranibizumab (40%) as well as in 2 control eyes in group 1. Same was detected in 1 eye injected with bevacizumab (20%) as well as in 1 control eye, in group 2. The noted atypical findings were lymphocytes and eosinophils in the vitreous posterior cavity and mild retinal inflammatory reaction with ganglion cell layer edema but without clinical significance. No atypical histopathological renal response was detected. CONCLUSION: Unexpected atypical histopathological retinal response without clinical significance was observed in 3 eyes injected with antiangiogenic drugs (2 in group 1 and 1 in group 2) as well as in 3 control eyes (2 in group 1 and 1 in group 2). No atypical renal response was detected suggesting no extra ocular involvement of the intravitreous injected antiangiogenic drugs.

An experimental model of chronic rhinosinusitis in rabbits without bacterial inoculation.

PURPOSE: Evaluate and compare two different experimental techniques of maxillary sinus ostium occlusion using N-butyl cyanoacrylate in developing chronic histological findings without the inoculation of pathogenic bacteria among rabbits. METHODS: In a randomized study, sixteen New Zealand rabbits were assigned for occlusion of the right maxillary sinus through a transmaxillary approach or through the roof of the nasal cavity. The contralateral sinus served as a control. After 12 weeks, the animals were sacrificed for blinded histopathological analysis of the maxillary sinus mucosa. RESULTS: Histopathological changes consistent with CRS were found in eight (100%) of the maxillary sinuses approached transmaxillary and three of those through the roof of the nasal cavity (37.5%), p 0.008 and 0.250, respectively, comparing with the control side. Chronic mucosal changes were significantly better induced using the transmaxillary approach (p 0.026). CONCLUSION: It is possible to induce a model of chronic sinusitis among rabbits with transmaxillary sinus occlusion without bacterial inoculation. This model can be replicated for future cellular studies.

An experimental model of chronic rhinosinusitis in rabbits without bacterial inoculation

PURPOSE: Evaluate and compare two different experimental techniques of maxillary sinus ostium occlusion using N-butyl cyanoacrylate in developing chronic histological findings without the inoculation of pathogenic bacteria among rabbits. METHODS: In a randomized study, sixteen New Zealand rabbits were assigned for occlusion of the right maxillary sinus through a transmaxillary approach or through the roof of the nasal cavity. The contralateral sinus served as a control. After 12 weeks, the animals were sacrificed for blinded histopathological analysis of the maxillary sinus mucosa. RESULTS: Histopathological changes consistent with CRS were found in eight (100%) of the maxillary sinuses approached transmaxillary and three of those through the roof of the nasal cavity (37.5%), p 0.008 and 0.250, respectively, comparing with the control side. Chronic mucosal changes were significantly better induced using the transmaxillary approach (p 0.026). CONCLUSION: It is possible to induce a model of chronic sinusitis among rabbits with transmaxillary sinus occlusion without bacterial inoculation. This model can be replicated for future cellular studies. .

Adequacy of ovarian diathermy under ultrasound control: an experimental model.

BACKGROUND: To develop a minimally invasive ovarian cauterization technique under transvaginal ultrasound control and evaluate the safety and feasability of monopolar cauterization to cause ovarian injury using female cattle of reproductive age as an experimental model. METHOD: Experimental study in a university research center was performed. Eleven female bovines of reproductive age were submitted to monopolar transvaginal ovarian cauterization. The right ovary (RO) was punctured at four sites and 40 W was applied for 5 s at each point, resulting in a total of 800 J (Joules) of thermal energy. In the left ovary (LO), the procedure was similar, with the same time and 80 W, resulting in a thermal energy of 1600 J. Macroscopic and microscopic lesions were assessed. RESULTS: Of 22 ovaries punctured, 20 were cauterized and exhibited macroscopic and typical microscopic lesions. No lesions could be found in the needle path. The measures of the areas of microscopic electrocautery lesions calculated estimating a cylindrical volume showed a median of 1.12% in the right ovary and 1.65% in the left ovary. When the estimate was calculated by spherical shape, the medians were 1.77% in the right ovary and 3.06% in the left ovary. There was a statistically significant difference in these two estimates (sphere, p = 0.008; cylinder, p = 0.021). CONCLUSION: The experimental animal model described for transvaginal ultrasound-guided ovarian needle cauterization seems to be feasible. The ovaries were successfully cauterized without injuries in needle path and more energy resulted in significantly more thermal lesion. The safety and effectiveness of this technique, theoretically less invasive than current ovarian drilling methods, could be tested in anovulatory women with PCOS.