Search for cytotoxic agents in multiple Laurencia complex seaweed species (Ceramiales, Rhodophyta) harvested from the Atlantic Ocean with emphasis on the Brazilian State of Espírito Santo

The development of new anti-cancer drugs of algal origin represents one of the least explored frontiers in medicinal chemistry. In this regard, the diversity of micro- and macroalgae found in Brazilian coastal waters can be viewed as a largely untapped natural resource. In this report, we describe a comparative study on the cytotoxic properties of extracts obtained from the Laurencia complex: Laurencia aldingensis, L. catarinensis, L. dendroidea, L. intricata, L. translucida, L. sp, and Palisada flagellifera. All of these species were collected in the coastal waters of the State of Espírito Santo, Brazil. Four out of the twelve samples initially investigated were found to show significant levels of toxicity towards a model tumor cell line (human uterine sarcoma, MES-SA). The highest levels of cytotoxicity were typically associated with non-polar (hexane) algal extracts, while the lowest levels of cytotoxicity were found with the corresponding polar (methanol) extracts. In this report, we also describe a biological model currently in development that will not only facilitate the search for new anti-cancer drug candidates of algal origin, but also permit the identification of compounds capable of inducing the destruction of multi-drug resistant tumors with greater efficiency than the pharmaceuticals currently in clinical use.

Rastreamento anterior para câncer de colo uterino em mulheres com alterações citológicas ou histológicas

OBJETIVO: Analisar a história de rastreamento citológico anterior em mulheres que apresentaram alterações citológicas e confirmação histológica para câncer cervical. MÉTODOS: Estudo transversal com 5.485 mulheres (15-65 anos) que se submeteram a rastreamento para o câncer cervical entre fevereiro de 2002 a março de 2003, em São Paulo e Campinas, SP. Aplicou-se questionário comportamental e foi feita a coleta da citologia oncológica convencional ou em base líquida. Para as participantes com alterações citológicas indicou-se colposcopia e, nos casos anormais, procedeu-se à biópsia cervical. Para investigar a associação entre as variáveis qualitativas e o resultado da citologia, utilizou-se o teste de qui-quadrado de Pearson com nível de significância de 5%. RESULTADOS: Dentre os resultados citológicos, 354 (6,4%) foram anormais, detectando-se 41 lesões intra-epitelial escamosa de alto grau e três carcinomas; em 92,6% revelaram-se normais. De 289 colposcopias realizadas, 145 (50,2%) apresentaram alterações. Dentre as biópsias cervicais foram encontrados 14 casos de neoplasia intra-epitelial cervical grau 3 e quatro carcinomas. Referiram ter realizado exame citológico prévio: 100% das mulheres com citologia compatível com carcinoma, 97,6% das que apresentaram lesões intra-epiteliais de alto grau, 100% daquelas com confirmação histológica de carcinoma cervical, e 92,9% das mulheres com neoplasia intra-epitelial cervical grau 3. A realização de citologia anterior em período inferior a três anos foi referida, respectivamente, por 86,5% e 92,8% dessas participantes com alterações citológicas e histológicas. CONCLUSÕES: Entre as mulheres que apresentaram confirmação histológica de neoplasia intra-epitelial cervical grau 3 ou carcinoma e aquelas que não apresentaram alterações histológicas não houve diferença estatisticamente significante do número de exames citológicos realizados, bem como o tempo do último exame citológico anterior.

Estudo randômico da correção cirúrgica do prolapso uterino através de tela sintética de polipropileno tipo I comparando histerectomia versus preservação uterina

OBJETIVO: Comparar os resultados anatômicos pós-operatórios de pacientes portadoras de prolapso uterino tratadas utilizando tela de polipropileno para correção dos defeitos do assoalho pélvico, comparando histerectomia vaginal com a preservação do útero. MÉTODO: Estudo randomizado com 31 mulheres portadoras de prolapso uterino estádio III ou IV (POP-Q) divididas em dois grupos: Grupo HV- 15 mulheres submetidas à histerectomia vaginal e reconstrução da anatomia do assoalho pélvico com tela de polipropileno tipo I (Nazca R-Promedon) e Grupo HP- 16 mulheres mulheres submetidas à reconstrução da anatomia do assoalho pélvico com tela de polipropileno tipo I (Nazca R-Promedon) preservando o útero. Raça, urgência miccional, constipação intestinal, dor sacral, sangramento e tempo de operação foram os parâmetros analisados. RESULTADOS: O tempo de seguimento médio foi de nove meses. Não se observou diferença entre os grupos nas complicações funcionais. O tempo cirúrgico foi 120 minutos para grupo HV versus 58.9 minutos para grupo HP ( p < 0.001 ) e o volume de perda sanguínea intraoperatória foi 120 mL no grupo HV versus 20 mL para grupo HP ( p < 0.001*). A taxa de sucesso objetivo foi 86.67% para grupo HV e 75% para grupo HP (p = 0,667). A taxa de erosão de tela foi 20% (3/15) de extrusão no grupo HV versus 18,75% (3/16) no grupo HP (p = 1,000). CONCLUSÃO: A correção cirúrgica do assoalho pélvico com telas nas portadoras de prolapso uterino apresentaram similaridade quer sendo ela feita com histeropexia quer com histerectomia. Contudo, o tempo cirúrgico e o volume da perda sanguínea foram significantemente maiores no grupo com histerectromia (HV). Operações vaginais com telas são procedimentos efetivos para a correção do prolapso.

Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO², >37ºC). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

Microbiological and histopathological aspects of canine pyometra

As pyometra is recognized as one of the main causes of disease and death in the bitch the purposes of this study were to evaluate microbiological and histopathological aspects of canine pyometra and to research the virulence factors of the E. coli isolates identifying possible risks to human health. The microbiological isolation from the intrauterine contents of 100 dogs with pyometra was carried out and the virulence factors in the E. coli strains were identified using PCR method. This study also consisted of the counting of microorganisms colonies forming units in samples of intrauterine content, tests of antimicrobial susceptibility of the E. coli isolates and the histological examination of the uterus. E. coli was the most prevalent microorganism isolated (76.6%) and 120 strains (79.5%) were positive for sfa, 86 (56.9%) were positive for cnf, 87 (57.6%) were positive for pap, 52 (34.4%) were positive for hly, 51 (33.8%) were positive for iuc and 5 (3.3%) were positive for afa genes. One observed more sensitivity of E. coli to norfloxacin, polimixin B, sulphazotrin, chloranfenicol and enrofloxacin. In 42% of the samples of uterine walls where microorganisms were isolated, the sizes of the areas of the inflammatory responses corresponded to 39-56%. Virulence factors were identified in 98.0% of the strains evaluated, demonstrating a high frequency of potentially pathogenic E. coli. It must be considered that dogs are animals that are living in close proximity to man for thousands of years and have an important role in the transmission of E. coli to other animals and to man.

Diferenciais intra-urbanos de peso ao nascer no município de São Paulo

OBJETIVO: Identificar diferenciais intra-urbanos e fatores de risco na prevalência de baixo peso ao nascer. MÉTODOS: Foram utilizadas as informações das declarações de nascido vivo de mães residentes no município de São Paulo, obtidos do Sistema de Informação de Nascidos Vivos e Fundação Seade, para o período de 2002 e 2003, totalizando 368.980 nascidos vivos. Os endereços foram geo-referenciados em setores censitários e classificados em seis grupos de vulnerabilidade segundo o Índice Paulista de Vulnerabilidade Social. Para identificação dos possíveis fatores de risco empregou-se análise de regressão logística. RESULTADOS: Observou-se tendência de crescimento da prevalência de baixo peso ao nascer com aumento da vulnerabilidade (de 6,8% a 8,1%). Houve diferenças significativas entre os grupos quanto às características maternas, assistência pré-natal e da proporção de nascimentos de não pré-termo de baixo peso. No grupo de baixo peso não pré-termo, proxy da presença de retardo do crescimento intra-uterino, residir em áreas vulneráveis (1,29;1,17-1,43) e características socioeconômicas maternas desaforáveis, como mães adolescentes (1,13;1,04-1,22), baixa escolaridade (1,26;1,17-1,35) e elevada paridade (1,10;1,01-1,20) foram fatores de risco, assim como mães idosas (1,38;1,30-1,47), e sem companheiro (1,15;1,11-1,20). A ausência de pré-natal apresentou o maior risco de baixo peso para nascimentos de pré-termo (3,39;2,86-4,02) e não pré-termo (2,12;1,87-2,41). Houve redução de risco de baixo peso com o aumento de consultas de pré-natal para nascimentos de pré-termo e não pré-termo. CONCLUSÕES: Há diferenças de prevalência de baixo peso ao nascer segundo grupos de vulnerabilidade. A assistência pré-natal mostrou-se desigual segundo grupos de vulnerabilidade e seu elevado risco para o baixo peso ao nascer indica a importância de ampliar o acesso e qualidade dos serviços de saúde

The relationship between birth weight and insulin resistance in childhood

Chronic diseases that are typical of adulthood may originate in intra-uterine life through inadequate fetal development. The present epidemiological cohort study of 506 healthy children aged 5\20138 years evaluated the relationship between birth weight and insulin resistance in an age group that has been assessed in few similar studies. Insulin concentration was determined by chemiluminescence and insulin resistance by the homeostasis model assessment (HOMA). Blood glucose, total cholesterol and fractions (LDL cholesterol and HDL cholesterol) and TAG concentrations were determined by automated enzymatic methods. Linear regression analysis investigated the relationship between birth weight (assessed as a continuous variable and in three categories: small for gestational age, SGA; adequate for gestational age and large for gestational age) and the HOMA index, using backward stepwise selection and biological models to explain the causal pathway of the relationship. There were negativeassociations between birth weight (P < 0·001), SGA (P = 0·027) and the HOMA index, and a positive association between waist circumference (P < 0·001) and the HOMA index. Considering the significant associations between birth weight and waist circumference (P < 0·001) and waist circumference and insulin resistance (P < 0·001), we can probably suspect that lower birth weight is a common cause of higher waist circumference and insulin resistance. In summary, the results of the present study showed increased insulin resistance in apparently healthy, young children, who had lower weight at birth and higher measurements of waist circumference. There is a need to develop public health policies that adopt preventive measures to promote adequate maternal-fetal and child development and enable early diagnosis of metabolic abnormalities

Prevalência do HPV em mulheres rastreadas para o câncer cervical

OBJETIVO: Analisar a prevalência da infecção genital por papilomavírus humano (HPV) de alto risco por faixa etária e fatores associados. MÉTODOS: Estudo transversal com amostra de 2.300 mulheres (15-65 anos) que buscaram rastreamento para o câncer cervical entre fevereiro de 2002 e março de 2003 em São Paulo e Campinas, estado de São Paulo. Aplicou-se questionário epidemiológico e realizou-se coleta cervical para citologia oncológica e teste de captura híbrida II. As análises estatísticas empregadas foram teste de qui-quadrado de Pearson e análise multivariada pelo método forward likelihood ratio. RESULTADOS: A prevalência total da infecção genital por HPV de alto risco foi de 17,8%, distribuída nas faixas etárias: 27,1% (<25 anos), 21,3% (25-34 anos), 12,1% (35-44 anos), 12,0% (45-54 anos) e de 13,9% (55-65 anos). Participantes com maior número de parceiros sexuais durante a vida apresentaram maior freqüência da infecção. Relacionamento estável, idade de 35 a 44 anos e ex-fumantes foram associados à proteção da infecção. A infecção genital por HPV de alto risco ocorreu em 14,3% das citologias normais, em 77,8% das lesões escamosas de alto grau e nos dois (100%) casos de carcinoma. CONCLUSÕES: A prevalência da infecção genital por HPV de alto risco na amostra estudada foi alta. Houve predomínio de casos abaixo dos 25 anos e tendência a um novo aumento após os 55 anos, com maior freqüência naqueles com maior número de parceiros sexuais durante a vida

Hemangiossarcoma primário intrauterino em um macaco aranha de cara vermelha (Ateles paniscus)

Hemangiosarcoma is a common neoplasm in dogs and less frequently seen in cats. In nonhuman primates, this tumor is rarely reported. A 17 year-old female spider monkey (Ateles paniscus) was submitted an ultrasound exam due to gestation suspicion, which was seen a circular mass intra-uterine measuring 1.3 cm. New exams shown increase of the mass to 4.4 x 3.0 cm associated with a viable fetus. Was realized cesarean with ovariohysterectomy and excision of the mass; however the animal died in less than 24 hours after the surgery. In the necropsy, severe hemoabdomen was evidenced, although the surgical stumps were properly ligated and the complete sutures. Macroscopically, the uterine mass was soft, dark heterogeneous and measuring 5.0 cm in diameter. Histologically was visualized proliferation of spindle cells that form vascular channels replete of erythrocytes and some with thrombus, marked pleomorphism, nucleolus evident, binucleated cells and mitotic figures were rare. The immunohistochemistry (IHC) was performed, using streptavidin-biotin peroxidase technique (Dako Cytomation, USA) with the use of antibodies CD31 (clone JC/70A), CD34 (clone QB-END/10). The IHC showed a specific antigen-antibody reaction for CD31. According to localization, morphology and IHC, the present study reports a primary uterine hemangiosarcoma in a spider monkey that caused hemostatic abnormalities and consequent death of the animal.

Recoleta uterina como estratégia para aumentar a taxa de embriões em fêmeas bovinas de corte e leite

Embryo recovery rate in superovulated cows after uterine flushing is lower than the ovulation rate, which contributes to the relative inefficiency of the conventional cervical recovery procedure. A simple and easy alternative to improve embryo recovery rate is the uterine re-flushing procedure. To evaluate the effect of re-flushing and the influence of breed and operator on bovine embryo recovery rate, 38 Nelore and 19 Jersey females were stimulated using FSH, with embryos being collected by one of two trained operators. At the end of flushing, the catheter was sealed and maintained into the uterine body, filled with flushing medium, while females were released to a paddock for 30 to 50 min, to be submitted to the re-flushing procedure by the same operator. A total of 599 structures were recovered out of 57 procedures, from which 423 (70.6%) were obtained in the first flushing and 176 (29.4%) after re-flushing. Mean recovery rates of 7.4 and 3.1 structures were obtained after the first and second flushing, respectively, for a total of 10.5 structures per cow. Structures were obtained in 73.6% ( 42 out of 57) of the re-flushing procedures. No breed effect was observed on total ova or embryo recovery, with 10.9 total ova collected from Jersey and 10.3 from Nelore females. Likewise, the embryo recovery rate obtained following uterine flushing or re-flushing did not differ either between Jersey (8.1 and 2.7) and Nelore (7.0 and 3.2) animals, or between operators A (7.5 and 3.3) and B (7.3 and 2.9), respectively. In conclusion, the uterine re-flushing procedure significantly increased the rate of embryo recovery in Jersey and Nelore females, with no operator influence being observed.

Ureteroscopy in Pregnant Women for Ureteral Stone

Introduction: The occurrence of urolithiasis in pregnancy represents a challenge in both diagnosis and treatment of this condition, because it presents risks not only to the mother but also to the fetus. Surgical treatment may be indicated for patients with infection, persistent pain, and obstruction of a solitary kidney. We present our experience on the management of pregnant patients with ureteral calculi and a review of the literature. Materials and Methods: The charts of 19 pregnant patients with obstructive ureteral calculi were retrospectively reviewed. Gestational age ranged from 13 to 33 weeks. In all patients, ureteral stone was diagnosed on abdominal ultrasound. In regard to localization, 15 calculi were in the distal ureter, 3 in the proximal ureter, and 1 in the interior of an ureterocele. Calculi size ranged from 6 to 10 mm (mean, 8 mm). The following criteria were used to indicate ureteroscopy: persistent pain with no improvement after clinical treatment, increase in renal dilation, or presence of uterine contractions. Nine patients (47.3%) were submitted to ureteroscopy. All calculi (100%) were removed with a stone basket extractor under continuous endoscopic vision. None of the calculi demanded the use of a lithotriptor. Results: Nine patients (47.3%) treated with clinical measurements presented no obstetric complications and spontaneous elimination of the calculi. Nine patients (47.3%) submitted to ureteroscopy had no surgical complications. There was remission of pain in all cases after ureteroscopy and ureteral catheter placement. Conclusion: The diagnosis and treatment of ureteral lithiasis in pregnant women present potential risks for the fetus and the mother. Conservative management is the first option, but ureteroscopy may be performed with safety and high success rates.

Maternal diabetes affects cell proliferation in developing rat placenta

Placentation starts with the formation of a spheroidal trophoblastic shell surrounding the embryo, thus facilitating both implantation into the uterine stroma and contact with maternal blood. Although it is known that diabetes increases the placental size and weight, the mechanisms responsible for this alteration are still poorly understood. In mammals, cellular proliferation occurs in parallel to placental development and it is possible that diabetes induces abnormal uncontrolled cell proliferation in the placenta similar to that seen in other organs (e.g. retina). To test this hypothesis, the objective of this work was to determine cell proliferation in different regions of the placenta during its development in a diabetic rat model. Accordingly, diabetes was induced on day 2 of pregnancy in Wistar rats by a single injection of alloxan (40 mg/kg i.v.). Placentas were collected on days 14, 17, and 20 postcoitum. Immunoperoxidase was used to identify Ki67 nuclear antigen in placental sections. The number of proliferating cells was determined in the total placental area as well as in the labyrinth, spongiotrophoblast and giant trophoblast cell regions. During the course of pregnancy, the number of Ki67 positive cells decreased in both control and diabetic rat placentas. However, starting from day 17 of pregnancy, the number of Ki67 positive cells in the labyrinth and spongiotrophoblast regions was higher in diabetic rat placentas as compared to control. The present results demonstrate that placentas from the diabetic rat model have a significantly higher number of proliferating cells in specific regions of the placenta and at defined developmental stages. It is possible that this increased cell proliferation promotes thickness of the placental barrier consequently affecting the normal maternal-fetal exchanges.

Modulation of small leucine-rich proteoglycans (SLRPs) expression in the mouse uterus by estradiol and progesterone

Background: We have previously demonstrated that four members of the family of small leucine-rich-proteoglycans (SLRPs) of the extracellular matrix (ECM), named decorin, biglycan, lumican and fibromodulin, are deeply remodeled in mouse uterine tissues along the estrous cycle and early pregnancy. It is known that the combined action of estrogen (E2) and progesterone (P4) orchestrates the estrous cycle and prepares the endometrium for pregnancy, modulating synthesis, deposition and degradation of various molecules. Indeed, we showed that versican, another proteoglycan of the ECM, is under hormonal control in the uterine tissues. Methods: E2 and/or medroxiprogesterone acetate (MPA) were used to demonstrate, by real time PCR and immunoperoxidase staining, respectively, their effects on mRNA expression and protein deposition of these SLRPs, in the uterine tissues. Results: Decorin and lumican were constitutively expressed and deposited in the ECM in the absence of the ovarian hormones, whereas deposition of biglycan and fibromodulin were abolished from the uterine ECM in the non-treated group. Interestingly, ovariectomy promoted an increase in decorin, lumican and fibromodulin mRNA levels, while biglycan mRNA conspicuously decreased. Hormone replacement with E2 and/or MPA differentially modulates their expression and deposition. Conclusions: The patterns of expression of these SLRPs in the uterine tissues were found to be hormone-dependent and uterine compartment-related. These results reinforce the existence of subpopulations of endometrial fibroblasts, localized into distinct functional uterine compartments, resembling the organization into basal and functional layers of the human endometrium.

Stromal cells play a role in cervical cancer progression mediated by MMP-2 protein

Metalloproteinases, especially metal loprotemase-2 (MMP-2), are known for their role in the degradation of the extracellular matrix. Nevertheless, a thorough understanding of MMP-2 expression in neoplastic lesions of the uterine cervix has yet to be accomplished. This study aimed to analyze the MMP-2 expression in cervical intraepithelial neoplasia III (CIN3) and in cervical squamous cell carcinoma, in tumor cells and adjacent stromal cells. MMP-2 expression was assessed by an immunohistochernical technique. MMP-2 expression was greater in the stromal cells of invasive carcinomas than in CIN3 (p < 0.0001). MMP-2 expression in stromal cells correlates with the clinical stage, gradually increasing as the tumor progresses (p = 0.04). This study corroborates that stromal cells play an important role in tumor invasion and progression, mediated by the progressive enhancement of MMP-2 expression from CIN3 to advanced invasive tumor. The intense MMP-2 expression most probably is associated with poor tumor prognosis.

Histopathological Changes in the Placentas and Fetuses of Mice Infected with Trypanosoma cruzi Isolated from the Myotis nigricans nigricans Bat

Histopathological changes and placental transmission were studied in the late stages of pregnancy in mice infected with a strain of Trypanosoma cruzi, isolated from a Myolis nigricans nigricans bat. Large amastigote nests were observed in uterine muscles, as well as in decidual and endothelial placental cells. In addition, persistent coagulative and fibrotic Vascular degeneration was observed. Large amastigote burdens were found in giant cells, spongioblasts and endothelial cells within the labyrinthine layer. Transplacental transmission was confirmed in 30% of the fetuses examined, in which amastrigote nests were seen only in striated muscle. During tire acute phase, intrauterine development was impaired as the result of parasitic invasion of the placenta, and fetal mortality rose to 10%. (C) 2008 Elsevier Ltd. All rights reserved.