The materno-fetal interface in llama (Lama guanicoe glama)

Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5mm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

The materno-fetal interface in llama (Lama guanicoe glama)

Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5μm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

Morfologia da placenta e interação materno-fetal em jumentas (Equus asinus) da raça Pêga

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Materno-fetal nutrient transfer across primary human trophoblast layer.

MATERNO-FETAL NUTRIENT TRANSFER ACROSS PRIMARY HUMAN TROPHOBLAST MONOLAYER Objectives: Polarized trophoblasts represent the transport and metabolic barrier between the maternal and fetal circulation. Currently human placental nutrient transfer in vitro is mainly investigated unidirectionallyon cultured primary trophoblasts, or bidirectionally on the Transwell® system using BeWo cells treated with forskolin. As forskolin can induce various gene alterations (e.g. cAMP response element genes), we aimed to establish a physiological primary trophoblast model for materno-fetal nutrient exchange studies without forskolin application. Methods: Human term cytotrophoblasts were isolated by enzymatic digestion and Percoll® gradient separation. The purity of the primary cells was assessed by flow cytometry using the trophoblast-specific marker cytokeratin-7. After screening different coating matrices, we optimized the growth conditions for the primary cytotrophoblasts on Transwell/ inserts. The morphology of 5 days cultured trophoblasts was determined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Membrane makers were visualized using confocal microscopy. Additionally transport studies were performed on the polarized trophoblasts in the Transwell® system. Results: During 5 days culture, the trophoblasts (>90% purity) developed a modest trans-epithelial electrical resistance (TEER) and a sizedependent apparent permeability coefficient (Papp) to fluorescently labeled compounds (MW ~400-70’000D). SEM analyses confirmed a confluent trophoblast layer with numerous microvilli at day six, and TEM revealed a monolayer with tight junctions. Immunocytochemistry on the confluent trophoblasts showed positivity for the cell-cell adhesion molecule E-cadherin, the tight junction protein ZO-1, and the membrane proteins ABCA1 and Na+/K+-ATPase. Vectorial glucose and cholesterol transport studies confirmed functionality of the cultured trophoblast barrier. Conclusion: Evidence from cell morphology, biophysical parameters and cell marker expressions indicate the successful and reproducible establishment of a primary trophoblast monolayer model suitable for transport studies. Application of this model to pathological trophoblasts will help to better understand the mechanism underlying gestational diseases, and to define the consequences of placental pathology on materno-fetal nutrient transport.

Determinants of mother-baby relationship evaluated during pregnancy

ABSTRACT Background Mental health promotion is supported by a strong body of knowledge and is a matter of public health with the potential of a large impact on society. Mental health promotion programs should be implemented as soon as possible in life, preferably starting during pregnancy. Programs should focus on malleable determinants, introducing strategies to reduce risk factors or their impact on mother and child, and also on strengthening protective factors to increase resilience. The ambition of early detecting risk situations requires the development and use of tools to assess risk, and the creation of a responsive network of services based in primary health care, especially maternal consultation during pregnancy and the first months of the born child. The number of risk factors and the way they interact and are buffered by protective factors are relevant for the final impact. Maternal-fetal attachment (MFA) is not yet a totally understood and well operationalized concept. Methodological problems limit the comparison of data as many studies used small size samples, had an exploratory character or used different selection criteria and different measures. There is still a lack of studies in high risk populations evaluating the consequences of a weak MFA. Instead, the available studies are not very conclusive, but suggest that social support, anxiety and depression, self-esteem and self-control and sense of coherence are correlated with MFA. MFA is also correlated with health practices during pregnancy, that influence pregnancy and baby outcomes. MFA seems a relevant concept for the future mother baby interaction, but more studies are needed to clarify the concept and its operationalization. Attachment is a strong scientific concept with multiple implications for future child development, personality and relationship with others. Secure attachment is considered an essential basis of good mental health, and promoting mother-baby interaction offers an excellent opportunity to intervention programmes targeted at enhancing mental health and well-being. Understanding the process of attachment and intervening to improve attachment requires a comprehension of more proximal factors, but also a broader approach that assesses the impact of more distal social conditions on attachment and how this social impact is mediated by family functioning and mother-baby interaction. Finally, it is essential to understand how this knowledge could be translated in effective mental health promoting interventions and measures that could reach large populations of pregnant mothers and families. Strengthening emotional availability (EA) seems to be a relevant approach to improve the mother-baby relationship. In this review we have offered evidence suggesting a range of determinants of mother-infant relationship, including age, marital relationship, social disadvantages, migration, parental psychiatric disorders and the situations of abuse or neglect. Based on this theoretical background we constructed a theoretical model that included proximal and distal factors, risk and protective factors, including variables related to the mother, the father, their social support and mother baby interaction from early pregnancy until six months after birth. We selected the Antenatal Psychosocial Health Assessment (ALPHA) for use as an instrument to detect psychosocial risk during pregnancy. Method Ninety two pregnant women were recruited from the Maternal Health Consultation in Primary Health Care (PHC) at Amadora. They had three moments of assessment: at T1 (until 12 weeks of pregnancy) they filed out a questionnaire that included socio-demographic data, ALPHA, Edinburgh post-natal Depression Scale (EDPS), General Health Questionnaire (GHQ) and Sense of Coherence (SOC); at T2 (after the 20th weeks of pregnancy) they answered EDPS, SOC and MFA Scale (MFAS), and finally at T3 (6 months after birth), they repeated EDPS and SOC, and their interaction with their babies was videotaped and later evaluated using EA Scales. A statistical analysis has been done using descriptive statistics, correlation analysis, univariate logistic regression and multiple linear regression. Results The study has increased our knowledge on this particular population living in a multicultural, suburb community. It allow us to identify specific groups with a higher level of psychosocial risk, such as single or divorced women, young couples, mothers with a low level of education and those who are depressed or have a low SOC. The hypothesis that psychosocial risk is directly correlated with MFAS and that MFA is directly correlated with EA was not confirmed, neither the correlation between prenatal psychosocial risk and mother-baby EA. The study identified depression as a relevant risk factor in pregnancy and its higher prevalence in single or divorced women, immigrants and in those who have a higher global psychosocial risk. Depressed women have a poor MFA, and a lower structuring capacity and a higher hostility to their babies. In average, depression seems to reduce among pregnant women in the second part of their pregnancy. The children of immigrant mothers show a lower level of responsiveness to their mothers what could be transmitted through depression, as immigrant mothers have a higher risk of depression in the beginning of pregnancy and six months after birth. Young mothers have a low MFA and are more intrusive. Women who have a higher level of education are more sensitive and their babies showed to be more responsive. Women who are or have been submitted to abuse were found to have a higher level of MFA but their babies are less responsive to them. The study highlights the relevance of SOC as a potential protective factor while it is strongly and negatively related with a wide range of risk factors and mental health outcomes especially depression before, during and after pregnancy. Conclusions ALPHA proved to be a valid, feasible and reliable instrument to Primary Health Care (PHC) that can be used as a total sum score. We could not prove the association between psychosocial risk factors and MFA, neither between MFA and EA, or between psychosocial risk and EA. Depression and SOC seems to have a clear and opposite relevance on this process. Pregnancy can be considered as a maturational process and an opportunity to change, where adaptation processes occur, buffering risk, decreasing depression and increasing SOC. Further research is necessary to better understand interactions between variables and also to clarify a better operationalization of MFA. We recommend the use of ALPHA, SOC and EDPS in early pregnancy as a way of identifying more vulnerable women that will require additional interventions and support in order to decrease risk. At political level we recommend the reinforcement of Immigrant integration and the increment of education in women. We recommend more focus in health care and public health in mental health condition and psychosocial risk of specific groups at high risk. In PHC special attention should be paid to pregnant women who are single or divorced, very young, low educated and to immigrant mothers. This study provides the basis for an intervention programme for this population, that aims to reduce broad spectrum risk factors and to promote Mental Health in women who become pregnant. Health and mental health policies should facilitate the implementation of the suggested measures.

Moxibustión en la malposición fetal

Introducción: La incidencia de la presentación fetal podálica en España es del 3,8%. La moxibustión es parte integral de la milenaria medicina tradicional china. La técnica consiste en la colocación de un palillo incandescente de Artemisia vulgaris a unos pocos centímetros del punto de acupuntura Zhiyin (punto 67 del meridiano de vejiga, ubicado en la base externa de la uña del quinto dedo del pie). Los objetivos son: 1. Conocer el porcentaje de fetos que han rotado a cefálica mediante la moxibustión en gestantes que presenten una malposición fetal a partir de las 32 semanas de embarazo; 2. Identificar las complicaciones materno-fetales en la aplicación de la técnica. Sujetos: Se estudiaron 18 gestantes de más de 32 semanas de embarazo. Material y métodos: Mediante un estudio descriptivo de intervención, analizamos el porcentaje de fetos que rotaron a presentación cefálica. Resultados: Quince gestantes (83,9%) realizaron el tratamiento adecuadamente y las 3 restantes (16,7%) lo realizaron de manera ocasional y terminaron con una cesárea electiva por nalgas. Conclusiones: Parece razonable concluir que el cumplimiento del tratamiento influye en el tipo de parto (χ2= 12,600; gl= 1; p= 0,000), aunque no modifica el Apgar del recién nacido. Esta técnica se presenta como una alternativa económica, segura, sencilla y práctica para la versión fetal de la presentación podálica.

Trauma na gestante: análise da mortalidade materna e fetal

Foram analisadas retrospectivamente 26 pacientes gestantes traumatizadas, num período de nove anos. A média de idade foi 23,7 anos (16-42). A idade gestacional variou de dez a quarenta semanas (média 21,5 semanas); a maioria (46,1%) no segundo trimestre. O mecanismo predominante (65,3%) foi o trauma abdominal fechado por acidente automobilístico (atropelamento ou colisão). Na admissão, oito (30,7%) pacientes apresentavam alterações hemodinâmicas. Seis doentes (23,0%) apresentavam sangramento vaginal e, destas, quatro estavam hemodinamicamente normais. Analisamos a mortalidade materna, a mortalidade fetal e suas causas. Comparamos também a mediana dos valores do RTS e TRISS entre os grupos, sobrevida materno-fetal, sobrevida materna e óbito materno-fetal. Todas as gestantes admitidas com sangramento vaginal apresentaram óbito fetal. A mortalidade materna foi de 11,5%, por choque hemorrágico. A mortalidade fetal foi de 30,7%, sendo que 37,5% destes óbitos foram provocados pela morte materna. A principal causa de mortalidade fetal foi o descolamento de placenta (50,0%). Os índices de trauma, RTS e TRISS, foram significativamente menor (p=0,0025 e p<0,0001) no grupo óbito materno-fetal, porém esses índices não apresentaram valor prognóstico na mortalidade fetal.

Cardiopatia fetal e estratégias de enfrentamento

OBJETIVOS: Avaliar as estratégias de enfrentamento (coping) das gestantes frente ao diagnóstico de cardiopatia fetal. MÉTODOS: Foram entrevistadas 50 gestantes que receberam o diagnóstico de cardiopatia fetal. Para a coleta de dados utilizou-se uma entrevista semidirigida e o Inventário de Estratégia de Coping. A entrevista foi realizada, em média, 22 dias após terem recebido o diagnóstico. RESULTADOS: Ao investigar como se sentiam em relação ao bebê, 56,0% relataram preocupação e fragilidade, enquanto que as demais (44,0%) afirmaram estarem felizes e bem. As estratégias mais utilizadas pelas gestantes foram: resolução de problemas (73,0%), suporte social (69,1%), fuga/esquiva (62,7%), e a estratégia menos utilizada foi a de afastamento (17,3%). Constatou-se que as mulheres com companheiro, utilizaram mais a estratégia de resolução de problemas (p<0,05), assim como as que tinham entre 1 e 2 filhos (p<0,05). CONCLUSÕES: As estratégias de enfrentamento ativas, voltadas para a resolução de problemas e pela busca de suporte social, associadas à responsabilidade e à necessidade de cuidados específicos para a sobrevivência e o bem-estar do bebê, propiciaram uma relação mais próxima com a gestação, fortalecendo o vínculo materno-fetal.

Enfermedad neonatal temprana por Streptococcus agalactiae en una unidad de recién nacidos, factores de riesgo materno-fetales asociados a severidad y mortalidad

La colonización materna por estreptococo del grupo B (SGB) en países en desarrollo es del 4-20%; el 50% de sus hijos nacen colonizados y el 1-2% desarrollan enfermedad invasiva con alto riesgo de morbimortalidad y secuelas. Objetivo: determinar los factores de riesgo materno-fetales asociados a enfermedad severa y mortalidad neonatal por SGB en una unidad de recién nacidos. Materiales y métodos: se realizó un estudio observacional analítico de cohorte histórica durante un período de dos años. Se tomaron 11 casos con enfermedad invasiva y 15 controles. Se calcularon frecuencias absolutas y relativas, y se buscaron asociaciones mediante el cálculo del estadístico chi2. Resultados: los factores maternos predictivos para enfermedad por SGB incluyeron fiebre periparto mayor a 37,5 grados centígrados (p < 0,05), corioamnionitis y ruptura de membranas mayor a 18 horas (p < 0,05). Los factores de riesgo neonatal incluyeron prematurez (< 37 semanas) y bajo peso al nacer (< 2.500 gramos) (p < 0,05). Se encontró una mortalidad de 5 (45%). Conclusiones: hubo relación estadísticamente significativa entre la corioaminionitis materna, la ruptura de membranas mayor a 18 horas, la prematurez y el peso bajo al nacer con la severidad de la enfermedad y la mortalidad. La incidencia estimada de infección neonatal en la unidad de recién nacidos fue de 1,8 casos/1.000 nacidos vivos, y la de colonización materna fue de 4,3 casos/1.000 maternas. Se deben realizar más estudios en el país para establecer la incidencia real de enfermedad neonatal por SGB y hacer investigación sobre la costo-efectividad de las medidas de prevención.

A ultra-sonografia obstétrica e suas implicações na relação mãe-feto : impressões e sentimentos de gestantes com e sem diagnóstico de anormalidade fetal

O objetivo desta pesquisa foi investigar as impressões e sentimentos das gestantes sobre a ultra-sonografia obstétrica e suas implicações na relação mãe-feto, no contexto de normalidade e anormalidade fetal. Para tanto, foram realizados dois estudos. Participaram do primeiro estudo onze gestantes primíparas, com idades entre 18 e 35 anos e idades gestacional entre 11 e 24 semanas, que estavam sendo submetidas pela primeira vez à ultra-sonografia. Elas responderam a uma entrevista semi-estruturada e à Escala de Apego Materno-Fetal, antes e depois do exame. Análise de conteúdo qualitativa das entrevistas, mostrou que a ultra-sonografia foi vista com satisfação, além de tornar o bebê mais real e concreto, o que, em geral, intensificou os comportamentos de interação mãe-bebê e os sentimentos maternos. O Teste Wilcoxon revelou um aumento significativo no apego materno fetal após o exame. O segundo estudo contou com três gestantes com diagnóstico confirmado de anormalidade fetal, com idades entre 21 e 30 anos, e idades gestacionais entre 28 e 35 semanas. As participantes foram entrevistadas três meses depois da notícia do diagnóstico. Análise de conteúdo qualitativa das entrevistas revelou que a ultra-sonografia foi vista com ambivalência pelas gestantes que reconheceram tanto aspectos positivos como negativos do exame. Os resultados dos dois estudos indicam que a ultra-sonografia exerceu um impacto emocional importante nas gestantes influenciando a relação mãe-bebê, tanto no contexto de normalidade como de anormalidade fetal.

Expressão gênica e protéica de receptores Toll-like em células do sangue periférico materno de gestações normais e complicadas por prematuridade

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expressão gênica e protéica de receptores Toll-like em células do sangue periférico materno de gestações normais e complicadas por prematuridade

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação do volume e índices vasculares placentários pela ultrassonografia tridimensional em gestações com restrição grave de crescimento fetal

INTRODUÇÃO: A restrição de crescimento fetal (RCF) representa uma das principais complicações da gravidez e está associada a elevadas taxas de morbimortalidade perinatal. A frequência de desfechos desfavoráveis neonatais está diretamente relacionada à gravidade da RCF, sendo que os casos de pior evolução estão relacionados com peso abaixo do percentil 3. O mecanismo do crescimento fetal não está totalmente esclarecido, mas resulta da interação entre potencial genético de crescimento e fatores placentários, maternos e ambientais. Dentre os fatores etiológicos, o desenvolvimento anormal da placenta e a diminuição da perfusão uteroplacentária são as principais causas de RCF. Este estudo teve por objetivo avaliar volume e índices de vascularização placentários, por meio da ultrassonografia tridimensional (US3D), em gestações com RCF grave, e as correlações dos parâmetros placentários com valores de normalidade e dopplervelocimetria materno-fetal. MÉTODOS: Foram avaliadas 27 gestantes cujos fetos apresentavam peso estimado abaixo do percentil 3 para a idade gestacional. Por meio da US3D, utilizando-se a técnica VOCAL, foram mensurados o volume placentário (VP) e os índices vasculares: índice de vascularização (IV), índice de fluxo (IF) e índice de vascularização e fluxo (IVF). Os dados foram comparados com a curva de normalidade para a idade gestacional e peso fetal descrita por De Paula e cols. (2008, 2009). Desde que os volumes placentários variam durante a gravidez, os valores observados foram comparados com os valores esperados para a idade gestacional e peso fetal. Foram criados os índices volume observado/ esperado para a idade gestacional (Vo/e IG) e volume placentário observado/ esperado para o peso fetal (Vo/e PF). Os parâmetros placentários foram correlacionados com índice de pulsatilidade (IP) médio de (AUt) e IP de artéria umbilical (AU), e avaliados segundo a presença de incisura protodiastólica bilateral em AUt. RESULTADOS: Quando comparadas à curva de normalidade, as placentas de gestação com RCF grave apresentaram VP, IV, IF e IVF significativamente menores (p < 0,0001 para todos os parâmetros). Houve correlação inversa estatisticamente significante da média do PI de AUt com o Vo/e IG (r= -0,461, p= 0,018), IV (r= -0,401, p= 0,042) e IVF (r= -0,421, p= 0,048). No grupo de gestantes que apresentavam incisura protodiastólica bilateral de artérias uterinas, Vo/e IG (p= 0,014), Vo/e PF (p= 0,02) e IV (p= 0,044) foram significativamente mais baixos. Nenhum dos parâmetros placentários apresentou correlação significativa com IP de AU. CONCLUSÕES: Observou-se que o volume e os índices de vascularização placentários apresentam-se diminuídos nos fetos com RCF grave. IP médio de AUT apresenta correlação negativa com Vo/e IG, IV e IVF, e Vo/e IG, Vo/e PF e IV apresentaram-se reduzidos nos casos de incisura bilateral. Não houve correlação significativa dos parâmetros placentários com IP de AU

Successful outcome of a surgically treated giant retroperitoneal liposarcoma during pregnancy

Background Retroperitoneal liposarcomas occur more frequently between the fifth and seventh decades. Fortunately, these tumors are exceedingly rare in pregnancy, but when they occur, their management becomes even more challenging. Case A pregnant patient with a retroperitoneal liposarcoma was treated by complete surgical resection at 13 weeks of gestation. The patient gave birth at 37 weeks of gestation to a normal newborn and remains free of disease after a year. Conclusions Individualized approach should be taken according to gestational phase, tumor kinetics and overall patient`s condition. Any disturbance to materno-fetal equilibrium, should be an indication for urgent therapeutic approach.