Rab35 controls cilium length, function and membrane composition.

Rab and Arl guanine nucleotide-binding (G) proteins regulate trafficking pathways essential for the formation, function and composition of primary cilia, which are sensory devices associated with Sonic hedgehog (Shh) signalling and ciliopathies. Here, using mammalian cells and zebrafish, we uncover ciliary functions for Rab35, a multitasking G protein with endocytic recycling, actin remodelling and cytokinesis roles. Rab35 loss via siRNAs, morpholinos or knockout reduces cilium length in mammalian cells and the zebrafish left-right organiser (Kupffer's vesicle) and causes motile cilia-associated left-right asymmetry defects. Consistent with these observations, GFP-Rab35 localises to cilia, as do GEF (DENND1B) and GAP (TBC1D10A) Rab35 regulators, which also regulate ciliary length and Rab35 ciliary localisation. Mammalian Rab35 also controls the ciliary membrane levels of Shh signalling regulators, promoting ciliary targeting of Smoothened, limiting ciliary accumulation of Arl13b and the inositol polyphosphate 5-phosphatase (INPP5E). Rab35 additionally regulates ciliary PI(4,5)P2 levels and interacts with Arl13b. Together, our findings demonstrate roles for Rab35 in regulating cilium length, function and membrane composition and implicate Rab35 in pathways controlling the ciliary levels of Shh signal regulators.

Follicular Helper T Cells Are Major Human Immunodeficiency Virus-2 Reservoirs and Support Productive Infection.

Follicular helper T cells (Tfh), CD4 lymphocytes critical for efficient antibody responses, have been shown to be key human immunodeficiency virus (HIV)-1 reservoirs. Human immunodeficiency virus-2 infection represents a unique naturally occurring model for investigating Tfh role in HIV/acquired immune deficiency syndrome, given its slow rate of CD4 decline, low to undetectable viremia, and high neutralizing antibody titers throughout the disease course. In this study, we investigated, for the first time, Tfh susceptibility to HIV-2 infection by combining in vitro infection of tonsillar Tfh with the ex vivo study of circulating Tfh from HIV-2-infected patients. We reveal that Tfh support productive HIV-2 infection and are preferential viral targets in HIV-2-infected individuals.

Molecular and electrophysiological characterization of anion transport in Arabidopsis thaliana pollen reveals regulatory roles for pH, Ca2+ and GABA.

We investigated the molecular basis and physiological implications of anion transport during pollen tube (PT) growth in Arabidopsis thaliana (Col-0). Patch-clamp whole-cell configuration analysis of pollen grain protoplasts revealed three subpopulations of anionic currents differentially regulated by cytoplasmic calcium ([Ca2+ ]cyt ). We investigated the pollen-expressed proteins AtSLAH3, AtALMT12, AtTMEM16 and AtCCC as the putative anion transporters responsible for these currents. AtCCC-GFP was observed at the shank and AtSLAH3-GFP at the tip and shank of the PT plasma membrane. Both are likely to carry the majority of anion current at negative potentials, as extracellular anionic fluxes measured at the tip of PTs with an anion vibrating probe were significantly lower in slah3-/- and ccc-/- mutants, but unaffected in almt12-/- and tmem16-/- . We further characterised the effect of pH and GABA by patch clamp. Strong regulation by extracellular pH was observed in the wild-type, but not in tmem16-/- . Our results are compatible with AtTMEM16 functioning as an anion/H+ cotransporter and therefore, as a putative pH sensor. GABA presence: (1) inhibited the overall currents, an effect that is abrogated in the almt12-/- and (2) reduced the current in AtALMT12 transfected COS-7 cells, strongly suggesting the direct interaction of GABA with AtALMT12. Our data show that AtSLAH3 and AtCCC activity is sufficient to explain the major component of extracellular anion fluxes, and unveils a possible regulatory system linking PT growth modulation by pH, GABA, and [Ca2+ ]cyt through anionic transporters.

Changing Paradigms in the Initial Treatment of Ectopic Pregnancy at a University Hospital in Brazil.

OBJECTIVE: To evaluate the use of different treatment options for ectopic pregnancy and the frequency of severe complications in a university hospital. METHODS: Observational study with women with ectopic pregnancy admitted at UNICAMP Womens Hospital, Brazil, between 01/01/2000 and 12/31/2017. The outcome variables were the type of treatment (first choice) and the presence of severe complications. Independent variables were clinical and sociodemographic data. Statistical analysis was carried out by the Cochran-Armitage test, chi-square test, Mann-Whitney test and multiple Cox regression. RESULTS: In total 673 women were included in the study. The mean age was 29.0 years (± 6.1) and the mean gestational age was 7.7 (± 2.5). The frequency of surgical treatment decreased significantly over time (z = -4.69; p < 0.001). Conversely, there was a significant increase in the frequency of methotrexate treatment (z = 4.73; p < 0.001). Seventy-one women (10.5%) developed some type of severe complication. In the final statistical model, the prevalence of severe complications was higher in women who were diagnosed with a ruptured ectopic pregnancy at admission (PR = 2.97; 95%CI: 1.61-5.46), did not present with vaginal bleeding (PR = 2.45; 95%CI: 1.41-4.25), had never undergone laparotomy/laparoscopy (PR = 6.69; 95%CI: 1.62-27.53), had a non-tubal ectopic pregnancy (PR = 4.61; 95%CI: 1.98-10.74), and do not smoke (PR = 2.41; 95%CI: 1.08-5.36). CONCLUSION: there was a change in the first treatment option for cases of ectopic pregnancy in the hospital during the period of analysis. Factors inherent to a disease that is more difficult to treat are related to a higher frequency of severe complications.

OBJETIVO: Avaliar as diferentes opções de tratamento para gravidez ectópica e a frequencia de complicações graves em um hospital universitário. MéTODOS: Estudo observacional com mulheres com gravidez ectópica admitidas no Hospital da Mulher da UNICAMP, no Brasill, entre 01/01/2000 e 31/12/2017. As variáveis de desfecho foram o tipo de tratamento (primeira escolha) e a presença de complicações graves. As variáveis independents foram dados clínicos e sociodemográficos. A análise estatística foi realizada pelo teste de Cochran­Armitage, teste de qui-quadrado, teste de Mann­Whitney e Regressão de Cox Múltipla. RESULADOS: No total, 673 mulheres foram incluídas no estudo. A idade médica foi de 29.0 anos (± 6.1) e a idade gestacional media foi de 7.7 (± 2.5). A frequencia de tratamento cirúrgico diminuiu significativamente ao longo dos anos(z = -4.69; p < 0.001). Simultaneamente, houve um aumento da frequencia do tratamento clínico(z = 4.73; p < 0.001). Setenta e uma mulheres (10.5%) desenvolveram algum tipo de complicação grave. No modelo estatístico final, a prevalência de complicações graves foi maior nas mulheres que tiveram diagnóstico de gestação ectópica rota à admissão (PR = 2.97; 95%CI: 1.61­5.46), que não apresentaram sangramento vaginal (PR = 2.45; 95%CI: 1.41­4.25), sem antecedentes de laparotomia/laparoscopia (PR = 6.69; 95%CI: 1.62­27.53), com gravidez ectópica não-tubária (PR = 4.61; 95%CI: 1.98­10.74), e não tabagistas (PR = 2.41; 95%CI: 1.08­5.36). CONCLUSãO: Houve uma mudança na escolha do primeiro tratamento indicado nos casos de gravidez ectópica durante o período analisado. Os fatores inerentes a doença relacionados a maior dificuldade de tratamento foram associados a maior frequencia de complicações graves.

Adaptation of a Brazilian university hospital to clinical treatment of ectopic pregnancy: Lessons learned over 17 years.

OBJECTIVE: To describe and compare the annual success rates of medical treatment in the analyzed period and to evaluate the associated factors. METHODS: Retrospective study with 158 women with tubal pregnancy followed up over 17 years. Statistical analysis was performed using the Cochran-Armitage test, the χ2 test, Mann-Whitney test, and multiple logistic regression. RESULTS: The success rate was 47.4%. There was a trend of significant change in the success rate of clinical treatment over time (Z = 2.01, P = 0.044); it was associated to undergoing treatment between 2012 and 2017 (P = 0.028), the absence of abdominal pain (P = 0.020), receiving a higher dose of methotrexate (P < 0.001), and less time hospitalized (P < 0.001). In the final statistical model, we observed that receiving a higher dose of methotrexate (P = 0.025, odds ratio [OR] 1.03, 95% confidence interval [CI] 1.00-1.06), having a low serum ß-HCG concentration before treatment (P = 0.003, OR 0.87, 95% CI 0.79-0.95), and not having abdominal pain (P = 0.004, OR 4.26, 95% CI 1.61-11.28) were factors associated with a higher chance of successful clinical treatment for tubal pregnancy. CONCLUSION: A greater chance of success was observed among women undergoing clinical treatment from 2012 onwards, who used higher doses of methotrexate, were asymptomatic at admission, and had low concentrations of ß-hCG.

Experience of a Tertiary Service in the Treatment of Women with Cervical Pregnancy.

OBJECTIVE: Cervical pregnancy is challenging for the medical community, as it is potentially fatal. The treatment can be medical or surgical; however, there are no protocols that establish the best option for each case. The objective of the present study was to describe the cases of cervical pregnancy admitted to a tertiary university hospital over a period of 18 years. METHODS: A retrospective study based on a review of the medical records of all cervical pregnancies admitted to the Women's Hospital at Universidade Estadual de Campinas, Southeastern Brazil, from 2000 to 2018. RESULTS: We identified 13 cases of cervical pregnancy out of a total of 673 ectopic pregnancies; only 1 case was initially treated with surgery because of hemodynamic instability. Of the 12 cases treated conservatively, 7 were treated with single-dose intramuscular methotrexate, 1, with intravenous and intramuscular methotrexate, 1, with intravenous methotrexate, 1, with 2 doses of intramuscular methotrexate, and 2, with intra-amniotic methotrexate. Of these cases, one had a therapeutic failure that required a hysterectomy. Two women received blood transfusions. Four women required cervical tamponade with a Foley catheter balloon for hemostasis. There was no fatal outcome. CONCLUSION: Cervical pregnancy is a rare and challenging condition from diagnosis to treatment. Conservative treatment was the primary method of therapy used, with satisfactory results. In cases of increased bleeding, cervical curettage was the initial treatment, and it was associated with the use of a cervical balloon for hemostasis.

OBJETIVO: A gravidez ectópica cervical é um desafio para a comunidade médica, pois pode ser fatal. O tratamento pode ser clínico ou cirúrgico, mas não existem protocolos que estabeleçam a melhor opção para cada caso. O objetivo deste estudo foi descrever os casos de gravidez ectópica cervical internados em um hospital universitário terciário durante 18 anos. MéTODOS: Estudo retrospectivo com revisão de prontuários de todas as gestações ectópicas cervicais internadas no Hospital da Mulher da Universidade Estadual de Campinas de 2000 a 2018. RESULTADOS: Foram identificados treze casos de gestação ectópica cervical em um total de 673 gestações ectópicas; apenas 1 caso foi inicialmente tratado com cirurgia por causa de instabilidade hemodinâmica. Dos 12 casos tratados conservadoramente, 7 foram tratados com metotrexato por via intramuscular em dose única, 1, com metotrexato pelas vias intravenosa e intramuscular, 1, com metotrexato por via intravenosa, 1, com 2 doses de metotrexato por via intramuscular, e 2, com metotrexato por via intra-amniótica. Desses casos, um apresentou falha terapêutica, e realizou-se uma histerectomia. Duas mulheres receberam transfusões de sangue. Quatro mulheres necessitaram de tamponamento cervical com cateter balão de Foley para hemostasia. Não houve casos fatais. CONCLUSãO: A gravidez cervical é uma condição rara e desafiadora desde o diagnóstico até o tratamento. O tratamento conservador foi o principal método terapêutico utilizado, com resultados satisfatórios. Nos casos de sangramento aumentado, a curetagem cervical foi o tratamento inicial, e foi associada ao uso de balão cervical para hemostasia.

Calcium-regulated anion channels in the plasma membrane of Lilium longiflorum pollen protoplasts.

• Currents through anion channels in the plasma membrane of Lilium longiflorum pollen grain protoplasts were studied under conditions of symmetrical anionic concentrations by means of patch-clamp whole-cell configuration. • With Cl(-) -based intra- and extracellular solutions, three outward-rectifying anion conductances, I(Cl1) , I(Cl2) and I(Cl3) , were identified. These three activities were discriminated by differential rundown behaviour and sensitivity to 5-nitro-2-(phenylpropylamino)-benzoate (NPPB), which could not be attributed to one or more channel types. All shared strong outward rectification, activated instantaneously and displayed a slow time-dependent activation for positive potentials. All showed modulation by intracellular calcium ([Ca(2+) ](in) ), increasing intensity from 6.04 nM up to 0.5 mM (I(Cl1) ), or reaching a maximum value with 8.50 µM (I(Cl2) and I(Cl3) ). • After rundown, the anionic currents measured using NO(3) (-) -based solutions were indistinguishable, indicating that the permeabilities of the channels for Cl(-) and NO(3) (-) are similar. Additionally, unitary anionic currents were measured from outside-out excised patches, confirming the presence of individual anionic channels. • This study shows for the first time the presence of a large anionic conductance across the membrane of pollen protoplasts, resulting from the presence of Ca(2+) -regulated channels. A similar conductance was also found in germinated pollen. We hypothesize that these putative channels may be responsible for the large anionic fluxes previously detected by means of self-referencing vibrating probes.

The essential role of anionic transport in plant cells: the pollen tube as a case study.

Plasma membrane anion transporters play fundamental roles in plant cell biology, especially in stomatal closure and nutrition. Notwithstanding, a lot is still unknown about the specific function of these transporters, their specific localization, or molecular nature. Here the fundamental roles of anionic transport in plant cells are reviewed. Special attention will be paid to them in the control of pollen tube growth. Pollen tubes are extreme examples of cellular polarity as they grow exclusively in their apical extremity. Their unique cell biology has been extensively exploited for fundamental understanding of cellular growth and morphogenesis. Non-invasive methods have demonstrated that tube growth is governed by different ion fluxes, with different properties and distribution. Not much is known about the nature of the membrane transporters responsible for anionic transport and their regulation in the pollen tube. Recent data indicate the importance of chloride (Cl(-)) transfer across the plasma membrane for pollen germination and pollen tube growth. A general overview is presented of the well-known accumulated data in terms of biophysical and functional characterization, transcriptomics, and genomic description of pollen ionic transport, and the various controversies around the role of anionic fluxes during pollen tube germination, growth, and development. It is concluded that, like all other plant cells so far analysed, pollen tubes depend on anion fluxes for a number of fundamental homeostatic properties.

Factors Associated with Abortion Complications after the Implementation of a Surveillance Network (MUSA Network) in a University Hospital.

OBJECTIVE: To evaluate the factors associated with abortion complications following the implementation of the good-practice surveillance network Mujeres en Situación de Aborto (Women Undergoing Abortion, MUSA, in Spanish). METHODS: A cross-sectional study with women who underwent abortion due to any cause and in any age group at UNICAMP Women's Hospital (part of MUSA network), Campinas, Brazil, between July 2017 and Agust 2019. The dependent variable was the presence of any abortion-related complications during hospitalization. The independent variables were clinical and sociodemographic data. The Chi-square test, the Mann-Whitney test, and multiple logistic regression were used for the statistical analysis. RESULTS: Overall, 305 women were enrolled (mean ± standard deviation [SD] for age: 29.79 ± 7.54 years). The mean gestational age was 11.17 (±3.63) weeks. Accidental pregnancy occurred in 196 (64.5%) cases, 91 (29.8%) due to contraception failure. At least 1 complication was observed in 23 (7.54%) women, and 8 (34.8%) of them had more than 1. The most frequent complications were excessive bleeding and infection. The factors independently associated with a higher prevalence of complications were higher gestational ages (odds ratio [OR]: 1.22; 95% confidence interval [95%CI]: 1.09 to 1.37) and contraceptive failure (OR: 3.4; 95%CI: 1.32 to 8.71). CONCLUSION: Higher gestational age and contraceptive failure were associated with a higher prevalence of complications. This information obtained through the surveillance network can be used to improve care, particularly in women more susceptible to unfavorable outcomes.

OBJETIVO: Avaliar os fatores associados às complicações em casos de aborto após a implementação da rede de vigilância de boas práticas Mujeres en Situación de Aborto (Mulheres em Situação de Aborto, MUSA, em espanhol). MéTODOS: Um estudo transversal, com mulheres admitidas por aborto de qualquer causa e em qualquer faixa etária, no Hospital da Mulher da UNICAMP (parte da rede MUSA), Campinas, Brasil, entre julho de 2017 e agosto de 2019. A variável dependente foi a presença de qualquer complicação relacionada ao quadro de aborto durante a hospitalização. As variáveis independentes foram dados clínicos e sociodemográficos. O teste de qui-quadrado, o teste de Mann-Whitney, e a regressão logística múltipla foram usados na análise estatística. RESULTADOS: Foram incluídas 305 mulheres (média ± desvio padrão [DP] da idade: 29,79 ± 7,54 anos). A idade gestacional média foi de 11,17 (±3,63) semanas. A gravidez não foi planejada em 196 (64,5%) casos, 91 (29,8%) devido a falha de contraceptivo. Pelo menos 1 complicação foi observada em 23 (7,64%) mulheres, 8 (34,8%) das quais apresentaram mais de uma complicação. As complicações mais frequentes foram sangramento excessivo e infecção. Os fatores independentemente associados à maior prevalência de complicações foram idades gestacionais maiores (razão de chances [OR]: 1.22; intervalo de confiança de 95% [IC95%]: 1.09 a 1.37) e falha de contraceptivo (OR: 3.4; IC95%: 1.32 a 8.71). CONCLUSãO: Maior idade gestacional e falha de contraceptivo estiveram associados à maior prevalência de complicações. As informações obtidas pela rede de vigilância podem ser usadas para melhorar o cuidado, particularmente nas mulheres mais suscetíveis a desfechos desfavoráveis.

Clinical Features and Management of Women with Borderline Ovarian Tumors in a Single Center in Brazil.

OBJECTIVE: The aim of the present study was to describe and analyze data of 57 women with borderline ovarian tumors (BOTs) regarding histological characteristics, clinical features and treatment management at the Department of Obstetrics and Gynecology of the Universidade Estadual de Campinas (Unicamp, in the Portuguese acronym). METHODS: The present retrospective study analyzed data obtained from clinical and histopathological reports of women with BOTs treated in a single cancer center between 2010 and 2018. RESULTS: A total of 57 women were included, with a mean age of 48.42 years old (15.43-80.77), of which 30 (52.63%) were postmenopausal, and 18 (31.58%) were < 40 years old. All of the women underwent surgery. A total of 37 women (64.91%) were submitted to complete surgical staging for BOT, and none (0/57) were submitted to pelvic or paraortic lymphadenectomy. Chemotherapy was administered for two patients who recurred. The final histological diagnoses were: serous in 20 (35.09%) cases, mucinous in 26 (45.61%), seromucinous in 10 (17.54%), and endometrioid in 1 (1.75%) case. Intraoperative analyses of frozen sections were obtained in 42 (73.68%) women, of which 28 (66.67%) matched with the final diagnosis. The mean follow-up was of 42.79 months (range: 2.03-104.87 months). Regarding the current status of the women, 45 (78.95%) are alive without disease, 2 (3.51%) are alive with disease, 9 (15.79%) had their last follow-up visit > 1 year before the performance of the present study but are alive, and 1 patient (1.75%) died of another cause. CONCLUSION: Women in the present study were treated according to the current guidelines and only two patients recurred.

OBJETIVO: O objetivo do presente estudo foi descrever uma série de 57 mulheres com tumores borderline de ovário (TBO) em relação às características histológicas, clínicas, e ao manejo do tratamento realizado no Departamento de Obstetrícia e Ginecologia da Universidade Estadual de Campinas (Unicamp). MéTODOS: O presente estudo retrospectivo analisou dados obtidos dos registros clínicos e histopatológicos de mulheres com TBO tratadas em um único centro oncológico de 2010 a 2018. RESULTADOS: Um total de 57 mulheres foram incluídas, com uma média de idade de 48,42 anos (15,43­80,77), das quais 30 (52,63%) eram menopausadas, e 18 (31,58%) tinham < 40 anos. Todas as mulheres foram operadas. Um total de 37 mulheres (64,91%) foram submetidas a cirurgia de estadiamento completo para TBO, e nenhuma foi submetida a linfadenectomia pélvica ou paraórtica. O tratamento com quimioterapia foi administrado em duas pacientes que recidivaram. Os diagnósticos histológicos finais foram: seroso em 20 mulheres (35,09%), mucinoso em 26 (45,61%), seromucinoso em 10 (17,54%) e endometrióide em 1 (1,75%). A avaliação histológica intraoperatória foi realizada em 42 (73,68%) das mulheres, das quais 28 (66,67%) foram compatíveis com os diagnósticos finais. O tempo médio de seguimento foi de 42,79 meses (variando de 2,03 a 104,87 meses). Em relação ao status atual das mulheres, 45 (78.95%) estão vivas sem doença, 2 (3,51%) estão vivas com doença, 9 (15.79%) tiveram a última consulta de seguimento há > 1 ano antes da realização do presente estudo, mas estão vivas, e 1 paciente faleceu por outra causa. CONCLUSãO: As mulheres do presente estudo foram tratadas de acordo com as recomendações atuais e apenas duas mulheres apresentaram recorrência.

Changing Paradigms in the Initial Treatment of Ectopic Pregnancy at a University Hospital in Brazil / Mudança de paradigmas do tratamento inicial de gravidez ectópica em um hospital universitário no Brasil

Abstract Objective To evaluate the use of different treatment options for ectopic pregnancy and the frequency of severe complications in a university hospital. Methods Observational study with women with ectopic pregnancy admitted at UNICAMP Womeńs Hospital, Brazil, between 01/01/2000 and 12/31/2017. The outcome variables were the type of treatment (first choice) and the presence of severe complications. Independent variables were clinical and sociodemographic data. Statistical analysis was carried out by the Cochran-Armitage test, chi-square test, Mann-Whitney test and multiple Cox regression. Results In total 673 women were included in the study. The mean age was 29.0 years (± 6.1) and the mean gestational age was 7.7 (± 2.5). The frequency of surgical treatment decreased significantly over time (z = -4.69; p < 0.001). Conversely, there was a significant increase in the frequency of methotrexate treatment (z = 4.73; p < 0.001). Seventy-one women (10.5%) developed some type of severe complication. In the final statistical model, the prevalence of severe complications was higher in women who were diagnosed with a ruptured ectopic pregnancy at admission (PR = 2.97; 95%CI: 1.61-5.46), did not present with vaginal bleeding (PR = 2.45; 95%CI: 1.41-4.25), had never undergone laparotomy/laparoscopy (PR = 6.69; 95%CI: 1.62-27.53), had a non-tubal ectopic pregnancy (PR = 4.61; 95%CI: 1.98-10.74), and do not smoke (PR = 2.41; 95%CI: 1.08-5.36). Conclusion there was a change in the first treatment option for cases of ectopic pregnancy in the hospital during the period of analysis. Factors inherent to a disease that is more difficult to treat are related to a higher frequency of severe complications.

Resumo Objetivo Avaliar as diferentes opções de tratamento para gravidez ectópica e a frequencia de complicações graves em um hospital universitário. Métodos Estudo observacional com mulheres com gravidez ectópica admitidas no Hospital da Mulher da UNICAMP, no Brasill, entre 01/01/2000 e 31/12/2017. As variáveis de desfecho foram o tipo de tratamento (primeira escolha) e a presença de complicações graves. As variáveis independents foram dados clínicos e sociodemográficos. A análise estatística foi realizada pelo teste de Cochran-Armitage, teste de qui-quadrado, teste de Mann-Whitney e Regressão de Cox Múltipla. Resulados No total, 673 mulheres foram incluídas no estudo. A idade médica foi de 29.0 anos (± 6.1) e a idade gestacional media foi de 7.7 (± 2.5). A frequencia de tratamento cirúrgico diminuiu significativamente ao longo dos anos(z = -4.69; p < 0.001). Simultaneamente, houve um aumento da frequencia do tratamento clínico(z = 4.73; p < 0.001). Setenta e uma mulheres (10.5%) desenvolveram algum tipo de complicação grave. No modelo estatístico final, a prevalência de complicações graves foi maior nas mulheres que tiveram diagnóstico de gestação ectópica rota à admissão (PR = 2.97; 95%CI: 1.61-5.46), que não apresentaram sangramento vaginal (PR = 2.45; 95%CI: 1.41-4.25), sem antecedentes de laparotomia/laparoscopia (PR = 6.69; 95%CI: 1.62-27.53), com gravidez ectópica não-tubária (PR = 4.61; 95%CI: 1.98-10.74), e não tabagistas (PR = 2.41; 95%CI: 1.08-5.36). Conclusão Houve uma mudança na escolha do primeiro tratamento indicado nos casos de gravidez ectópica durante o período analisado. Os fatores inerentes a doença relacionados a maior dificuldade de tratamento foram associados a maior frequencia de complicações graves.

Experience of a Tertiary Service in the Treatment of Women with Cervical Pregnancy / Experiência de um serviço terciário no tratamento de mulheres com gravidez ectópica cervical

Abstract Objective Cervical pregnancy is challenging for the medical community, as it is potentially fatal. The treatment can be medical or surgical; however, there are no protocols that establish the best option for each case. The objective of the present study was to describe the cases of cervical pregnancy admitted to a tertiary university hospital over a period of 18 years. Methods A retrospective study based on a review of the medical records of all cervical pregnancies admitted to the Women's Hospital at Universidade Estadual de Campinas, Southeastern Brazil, from 2000 to 2018. Results We identified 13 cases of cervical pregnancy out of a total of 673 ectopic pregnancies; only 1 case was initially treated with surgery because of hemodynamic instability. Of the 12 cases treated conservatively, 7 were treated with single-dose intramuscular methotrexate, 1, with intravenous and intramuscular methotrexate, 1, with intravenous methotrexate, 1, with 2 doses of intramuscular methotrexate, and 2, with intra-amniotic methotrexate. Of these cases, one had a therapeutic failure that required a hysterectomy. Two women received blood transfusions. Four women required cervical tamponade with a Foley catheter balloon for hemostasis. There was no fatal outcome. Conclusion Cervical pregnancy is a rare and challenging condition from diagnosis to treatment. Conservative treatment was the primary method of therapy used, with satisfactory results. In cases of increased bleeding, cervical curettage was the initial treatment, and it was associated with the use of a cervical balloon for hemostasis.

Resumo Objetivo A gravidez ectópica cervical é um desafio para a comunidade médica, pois pode ser fatal. O tratamento pode ser clínico ou cirúrgico, mas não existem protocolos que estabeleçam a melhor opção para cada caso. O objetivo deste estudo foi descrever os casos de gravidez ectópica cervical internados em um hospital universitário terciário durante 18 anos. Métodos Estudo retrospectivo com revisão de prontuários de todas as gestações ectópicas cervicais internadas no Hospital da Mulher da Universidade Estadual de Campinas de 2000 a 2018. Resultados Foram identificados treze casos de gestação ectópica cervical em um total de 673 gestações ectópicas; apenas 1 caso foi inicialmente tratado com cirurgia por causa de instabilidade hemodinâmica. Dos 12 casos tratados conservadoramente, 7 foram tratados com metotrexato por via intramuscular em dose única, 1, com metotrexato pelas vias intravenosa e intramuscular, 1, com metotrexato por via intravenosa, 1, com 2 doses de metotrexato por via intramuscular, e 2, com metotrexato por via intra-amniótica. Desses casos, um apresentou falha terapêutica, e realizou-se uma histerectomia. Duas mulheres receberam transfusões de sangue. Quatro mulheres necessitaram de tamponamento cervical com cateter balão de Foley para hemostasia. Não houve casos fatais. Conclusão A gravidez cervical é uma condição rara e desafiadora desde o diagnóstico até o tratamento. O tratamento conservador foi o principal método terapêutico utilizado, com resultados satisfatórios. Nos casos de sangramento aumentado, a curetagem cervical foi o tratamento inicial, e foi associada ao uso de balão cervical para hemostasia.

Notch/Her12 signalling modulates, motile/immotile cilia ratio downstream of Foxj1a in zebrafish left-right organizer.

Foxj1a is necessary and sufficient to specify motile cilia. Using transcriptional studies and slow-scan two-photon live imaging capable of identifying the number of motile and immotile cilia, we now established that the final number of motile cilia depends on Notch signalling (NS). We found that despite all left-right organizer (LRO) cells express foxj1a and the ciliary axonemes of these cells have dynein arms, some cilia remain immotile. We identified that this decision is taken early in development in the Kupffer's Vesicle (KV) precursors the readout being her12 transcription. We demonstrate that overexpression of either her12 or Notch intracellular domain (NICD) increases the number of immotile cilia at the expense of motile cilia, and leads to an accumulation of immotile cilia at the anterior half of the KV. This disrupts the normal fluid flow intensity and pattern, with consequent impact on dand5 expression pattern and left-right (L-R) axis establishment.

Factors Associated with Abortion Complications after the Implementation of a Surveillance Network (MUSA Network) in a University Hospital / Fatores associados a complicações do aborto após a implementação de um sistema de vigilância (Rede MUSA) em um hospital universitário

Abstract Objective To evaluate the factors associated with abortion complications following the implementation of the good-practice surveillance network Mujeres en Situación de Aborto (Women Undergoing Abortion, MUSA, in Spanish). Methods A cross-sectional study withwomen who underwent abortion due to any cause and in any age group at UNICAMP Women's Hospital (part of MUSA network), Campinas, Brazil, between July 2017 and Agust 2019. The dependent variable was the presence of any abortion-related complications during hospitalization. The independent variables were clinical and sociodemographic data. The Chi-square test, the Mann-Whitney test, and multiple logistic regression were used for the statistical analysis. Results Overall, 305 women were enrolled (mean±standard deviation [SD] for age: 29.79±7.54 years). The mean gestational age was 11.17 (±3.63) weeks. Accidental pregnancy occurred in 196 (64.5%) cases, 91 (29.8%) due to contraception failure. At least 1 complication was observed in 23 (7.54%) women, and 8 (34.8%) of them had more than 1. The most frequent complications were excessive bleeding and infection. The factors independently associated with a higher prevalence of complications were higher gestational ages (odds ratio [OR]: 1.22; 95% confidence interval [95%CI]: 1.09 to 1.37) and contraceptive failure (OR: 3.4; 95%CI: 1.32 to 8.71). Conclusion Higher gestational age and contraceptive failure were associated with a higher prevalence of complications. This information obtained through the surveillance network can be used to improve care, particularly in women more susceptible to unfavorable outcomes.

Resumo Objetivo Avaliar os fatores associados às complicações em casos de aborto após a implementação da rede de vigilância de boas práticas Mujeres en Situación de Aborto (Mulheres em Situação de Aborto, MUSA, em espanhol). Métodos Um estudo transversal, com mulheres admitidas por aborto de qualquer causa e em qualquer faixa etária, no Hospital da Mulher da UNICAMP (parte da rede MUSA), Campinas, Brasil, entre julho de 2017 e agosto de 2019. A variável dependente foi a presença de qualquer complicação relacionada ao quadro de aborto durante a hospitalização. As variáveis independentes foram dados clínicos e sociodemográficos. O teste de qui-quadrado, o teste de Mann-Whitney, e a regressão logística múltipla foram usados na análise estatística. Resultados Foram incluídas 305 mulheres (média±desvio padrão [DP] da idade: 29,79±7,54 anos). A idade gestacional média foi de 11,17 (±3,63) semanas. A gravidez não foi planejada em 196 (64,5%) casos, 91 (29,8%) devido a falha de contraceptivo. Pelo menos 1 complicação foi observada em 23 (7,64%) mulheres, 8 (34,8%) das quais apresentaram mais de uma complicação. As complicações mais frequentes foram sangramento excessivo e infecção. Os fatores independentemente associados à maior prevalência de complicações foram idades gestacionais maiores (razão de chances [OR]: 1.22; intervalo de confiança de 95% [IC95%]: 1.09 a 1.37) e falha de contraceptivo (OR: 3.4; IC95%: 1.32 a 8.71). Conclusão Maior idade gestacional e falha de contraceptivo estiveram associados à maior prevalência de complicações. As informações obtidas pela rede de vigilância podem ser usadas para melhorar o cuidado, particularmente nas mulheres mais suscetíveis a desfechos desfavoráveis.

Left-right organizer flow dynamics: how much cilia activity reliably yields laterality?

Internal organs are asymmetrically positioned inside the body. Embryonic motile cilia play an essential role in this process by generating a directional fluid flow inside the vertebrate left-right organizer. Detailed characterization of how fluid flow dynamics modulates laterality is lacking. We used zebrafish genetics to experimentally generate a range of flow dynamics. By following the development of each embryo, we show that fluid flow in the left-right organizer is asymmetric and provides a good predictor of organ laterality. This was tested in mosaic organizers composed of motile and immotile cilia generated by dnah7 knockdowns. In parallel, we used simulations of fluid dynamics to analyze our experimental data. These revealed that fluid flow generated by 30 or more cilia predicts 90% situs solitus, similar to experimental observations. We conclude that cilia number, dorsal anterior motile cilia clustering, and left flow are critical to situs solitus via robust asymmetric charon expression.

Integrated treatment between orthodontics and pediatric dentistry using the ulectomy technique: case report

Introduction: In the pediatric dental clinic the impacted teeth can be observedeventually, which can lead to the development of occlusion disorders. Objective:This study aimed to report a clinical case in which the surgical technique ofulectomy was used in conjunction with orthodontic treatment to aid the eruptionof a maxillary central incisor. Case Report: Male patient, 13 years old, Angle ClassI in the mixed dentition stage, with the tooth 21 impacted and reduced space forits eruption due to the migration of adjacent teeth. Fixed appliance was mountedwith Edgewise brackets using 0.20" stainless steel archwire with tight omega andactive spring between teeth 11 and 22 to recover the space of 21, but the passiveeruption was not observed. A fibrous gingival tissue was found, which by palpationrevealed the incisal edge of the tooth 21. The patient was referred to a pediatricdental clinic for a ulectomy. The minimally invasive surgical procedure consistedof the excision of the gingival tissue that covered the crown of the tooth 21 allowingits eruption in the dental arch. After the surgery, the teeth erupted and correctiveorthodontic treatment continued. Conclusion: The ulectomy technique associatedwith orthodontic treatment allowed to reestablish conditions for the developmentof dentition with satisfactory aesthetic and functional characteristics.

Introdução: Eventualmente na clínica odontológica, pode ser observada aimpacção de dentes, que pode acarretar transtornos para o desenvolvimento daoclusão. Objetivo: Objetivou-se relatar um caso clínico no qual a técnica cirúrgicade ulectomia foi utilizada, associada ao tratamento ortodôntico para auxiliar aerupção dentária de um incisivo central superior. Relato de Caso: Paciente dosexo masculino, 13 anos de idade, Classe I de Angle na fase de dentição mista, comdente 21 incluso e espaço reduzido para sua erupção em função da migração dosdentes adjacentes. Foi montado aparelho fixo com bráquetes prescrição Edgewise,utilizando arco 0,20" aço inoxidável com ômega justo e mola ativa entre osdentes 11 e 22 para recuperação do espaço do 21, mas não observou-se a erupçãopassiva. Constatou-se presença de tecido gengival fibroso que, ao toque,evidenciava a borda incisal do referido dente. O paciente foi encaminhado paraclínica de Odontopediatria para a realização da ulectomia. O procedimentocirúrgico minimamente invasivo, consistiu na exérese do tecido gengival querevestia a coroa do dente 21 não irrompido, permitindo sua erupção no arcodentário. Após a cirurgia, o dente irrompeu e o tratamento ortodôntico corretivoprosseguiu. Conclusão: A técnica da ulectomia associada ao tratamentoortodôntico permitiu restabelecer condições para desenvolvimento da dentiçãocom características estético-funcionais satisfatórias.

The importance of Zebrafish in biomedical research.

INTRODUCTION: Zebrafish (Danio rerio) is an ideal model organism for the study of vertebrate development. This is due to the large clutches that each couple produces, with up to 200 embryos every 7 days, and to the fact that the embryos and larvae are small, transparent and undergo rapid external development. MATERIAL AND METHODS: Using scientific literature research tools available online and the keywords Zebrafish, biomedical research, human disease, and drug screening, we reviewed original studies and reviews indexed in PubMed. RESULTS: In this review we summarized work conducted with this model for the advancement of our knowledge related to several human diseases. We also focused on the biomedical research being performed in Portugal with the zebrafish model. DISCUSSION: Powerful live imaging and genetic tools are currently available for zebrafish making it a valuable model in biomedical research. The combination of these properties with the optimization of automated systems for drug screening has transformed the zebrafish into "a top model" in biomedical research, drug discovery and toxicity testing. Furthermore, with the optimization of xenografts technology it will be possible to use zebrafish to aide in the choice of the best therapy for each patient. CONCLUSION: Zebrafish is an excellent model organism in biomedical research, drug development and in clinical therapy.

Introdução: O peixe-zebra (Danio rerio) é um excelente organismo modelo para o estudo do desenvolvimento dos vertebrados. Este facto deve-se às grandes ninhadas que cada casal produz, que podem atingir 200 embriões a cada sete dias, e ao facto dos embriões serem pequenos, transparentes e com um rápido desenvolvimento externo.Material e Métodos: Usando ferramentas de pesquisa bibliográfica científica disponíveis online e utilizando e as palavras-chave "Zebrafish", "biomedical research", "human disease" e "drug screening", avaliámos estudos originais e revisões indexadas na PubMed.Resultados: Neste artigo de revisão fazemos um resumo do trabalho realizado com este modelo no melhoramento doconhecimento de várias doenças humanas. Fizemos ainda um breve relato da investigação biomédica realizada em Portugal com o modelo de peixe-zebra.Discussão: Têm sido desenvolvidas poderosas ferramentas genéticas e de microscopia in vivo, que também tornaram o peixe-zebra num modelo valioso em investigação biomédica. A conjugação destes atributos com a optimização de sistemas automatizados de triagem de medicamentos, transformaram o peixe-zebra num top model da investigação em biomedicina, nomeadamente na triagem de compostos químicos com efeitos terapêuticos e em testes de toxicidade. Além disso, com a otimização da tecnologia dos xenografos, será possível usar o peixe-zebra na escolha de uma terapia personalizada.Conclusão: O peixe-zebra é um excelente organismo modelo na pesquisa biomédica, em screens de medicamentos e na terapia clinica.

IA1 microinvasive cervical adenocarcinoma in a young woman: conservative procedure and reproductive outcome / Adenocarcinoma microinvasor IA1 do colo do útero em mulher jovem: conduta conservadora e resultado reprodutivo

Introduction: The expansion of cytological screening programs for cervical cancer leads to an increase in the proportion of both adenocarcinomas and diagnoses in young women with reproductive intention. Conservative treatment is not fully established. Objective: to report the conservative management and follow-up difficulties of a real case of microinvasive cervical adenocarcinoma in a young woman. Case report: This is a case report of a 23-yearold patient with stage IA1 microinvasive cervical adenocarcinoma related to HPV18. The patient was vaccinated against HPV 16/18 at 16 years of age and conservatively treated. She became pregnant during follow-up with a favorable outcome. Conclusion: We discuss the difficulties and uncertainties regarding follow-up and opportunity for hysterectomy, emphasizing the need for a multidisciplinary approach to make a balanced decision between conservative treatment and oncological safety, as well as mitigate follow-up difficulties in real life.

Introdução: A ampliação dos programas de rastreamento citológico de câncer do colo uterino resulta em aumento na proporção de adenocarcinomas e de diagnósticos em jovens ainda com desejo reprodutivo. O tratamento conservador não está totalmente estabelecido. Objetivo: descrever a condução conservadora e as dificuldades de seguimento em caso real de adenocarcinoma microinvasor do colo em uma jovem. Relato de caso: É relatado o caso de uma paciente de 23 anos com adenocarcinoma microinvasor IA1 do colo uterino relacionado ao HPV 18, mesmo vacinada contra HPV 16/18 aos 16 anos. Foi tratada conservadoramente, com gestação e desfecho favorável. Conclusão: São discutidas as dificuldades e incertezas em relação ao seguimento e à oportunidade para histerectomia, ressaltando a necessidade de abordagem multidisciplinar para decisões equilibradas entre tratamento conservador, segurança oncológica e, ainda, amenizar as dificuldades de seguimento na vida real.

Perfil dos pacientes com úlcera crônica de etiologia diversa, internados em serviço de emergência / Perfil de los pacientes portadores de úlcera crónica de etiología diversa, internados en servicio de urgencia / Profile of patients diagnosed with chronic ulcer of diverse etiologyadmitted to an emergency unit

RESUMO Estudo com o objetivo de identificar o perfil dos pacientes com úlcera crônica de etiologia diversa internados em emergência hospitalar com Diagnóstico de Enfermagem de Integridade da Pele Prejudicada e de Integridade Tissular Prejudicada, segundo a Taxonomia da NANDA-I (2012-2014). Abordagem quantitativa, descritiva, de amostra intencional que analisou 41 prontuários de pacientes com úlcera crônica e comprometimento na integridade da pele. A coleta de dados ocorreu de setembro a Dezembro de 2013, por meio de busca ativa nos prontuários dos pacientes. Os dados foram submetidos à análise de estatística simples. Resultados apontam a predominância de pacientes do sexo masculino; idosos; com mobilidade prejudicada, baixa escolaridade, déficit de conhecimento de cuidados com a úlcera; família como principal rede de apoio. Destaca-se a incidência maior de úlcera arterial (65,9%), seguida pela úlcera por pressão (19,5%) e úlcera venosa (14,6%). Os problemas relacionados à integridade da pele têm relação com o aumento da expectativa de vida e com o desenvolvimento de doenças crônicas. Conhecer o perfil do paciente com úlcera crônica contribui para a elaboração de diretrizes para o cuidado de enfermagem e para o tratamento dessa condição de saúde.

RESUMEN estudio con el objetivo de identificar el perfil de los pacientes portadores de úlcera crónica de etiología diversa internados en urgencia hospitalaria con Diagnóstico de Enfermería de Integridad de la Piel Perjudicada y de Integridad Tisular Perjudicada, según la Taxonomía NANDA-I (2012-2014). Abordaje cuantitativo, descriptivo, de muestreo intencional que analizó 41 registros médicos de pacientes con úlcera crónica y comprometimiento en la integridad de la piel. La recolección de datos ocurrió de septiembre a diciembre de 2013, por medio de búsqueda activa en los registros médicos de los pacientes. Los datos fueron sometidos al análisis de estadística simple. Los resultados señalan el predominio de pacientes de sexo masculino; ancianos; con movilidad perjudicada, baja escolaridad, déficit de conocimiento de cuidados con la úlcera; familia como principal red de apoyo. Se destaca la incidencia mayor de la úlcera arterial (65,9%), seguida por la úlcera por presión (19,5%) y úlcera venosa (14,6%). Los problemas relacionados a la integridad de la piel tienen relación con el aumento de la expectativa de vida y con el desarrollo de enfermedades crónicas. Conocer el perfil del paciente portador de úlcera crónica contribuye para la elaboración de directrices para el cuidado de enfermería y para el tratamiento de esta condición de salud.

ABSTRACT Study aiming at identifying the profile of patients with chronic ulcer of diverse etiology admitted to an emergency unit, with Impaired Skin Integrity and Impaired Tissue Integrity Nursing Diagnoses, according to NANDA-I Taxonomy (2012-2014). Quantitative and descriptive approach of an intentional sample, which analyzed 41 medical records of patients with chronic ulcer and impaired skin integrity. Data were collected from September to December of 2013 by means of active search through the patients' records. Data were subjected to simple statistical analysis. Results evidence the predominance of patients with the following characteristics: males, elderly, impaired mobility, low education level, deficient knowledge about ulcer care, and family as main support network. It is worth highlighting the higher incidence of arterial ulcer (65.9%), followed by pressure ulcer (19.5%) and venous ulcer (14.6%). Problems related to skin integrity have a connection with increased life expectancy and the onset of chronic diseases. Learning the profile of patients with chronic ulcer contributes to the elaboration of guidelines targeting nursing care and the treatment of this health condition.