Robust deep learning optical autofocus system applied to automated multiwell plate single molecule localization microscopy.

We presenta robust, long-range optical autofocus system for microscopy utilizing machine learning. This can be useful for experiments with long image data acquisition times that may be impacted by defocusing resulting from drift of components, for example due to changes in temperature or mechanical drift. It is also useful for automated slide scanning or multiwell plate imaging where the sample(s) to be imaged may not be in the same horizontal plane throughout the image data acquisition. To address the impact of (thermal or mechanical) fluctuations over time in the optical autofocus system itself, we utilize a convolutional neural network (CNN) that is trained over multiple days to account for such fluctuations. To address the trade-off between axial precision and range of the autofocus, we implement orthogonal optical readouts with separate CNN training data, thereby achieving an accuracy well within the 600 nm depth of field of our 1.3 numerical aperture objective lens over a defocus range of up to approximately +/-100 µm. We characterize the performance of this autofocus system and demonstrate its application to automated multiwell plate single molecule localization microscopy.

Many microscopy experiments involve extended imaging of samples over timescales from minutes to days, during which the microscope can 'drift' out of focus. When imaging at high magnification, the depth of field is of the order of one micron and so the imaging system should keep the sample in the focal plane of the microscope objective lens to this precision. Unfortunately, temperature changes in the laboratory can cause thermal expansion of microscope components that can move the focal plane by more than a micron and such changes can occur on a timescale of minutes. This is a particular issue for super-resolved microscopy experiments using single molecule localization microscopy (SMLM) techniques, for which 1000s of images are acquired, and for automated imaging of multiple samples in multiwell plates. It is possible to maintain the sample in the focal plane focus position by either automatically moving the sample or adjusting the imaging system, for example by moving the objective lens. This is called 'autofocus' and is frequently achieved by reflecting a light beam from the microscope coverslip and measuring its position of beam profile as a function of defocus of the microscope. The correcting adjustment is then usually calculated analytically but there is recent interest in using machine learning techniques to determine the required focussing adjustment. Here, we present a system that uses a neural network to determine the required defocus correcting adjustment from camera images of a laser beam that is reflected from the coverslip. Unfortunately, this approach will only work when the microscope is in the same condition as it was when the neural network was trained - and this can be compromised by the same drift of the optical system that causes the defocus needing to be corrected. We show, however, that by training a neural network over an extended period, for example 10 days, this approach can 'learn' about the optical system drifts and provide the required autofocus function. We also show that an optical system utilizing a rectangular slit can make two measurements of the defocus simultaneously, with one measurement being optimized for high accuracy over a limited range (±10 µm) near focus and the other providing lower accuracy but over a much longer range (±100 µm). This robust autofocus system is suitable for automated super-resolved microscopy of arrays of samples in a multiwell plate using SMLM, for which an experiment routinely lasts more than 5 h.

Experience of oncology residents with death: a qualitative study in Mexico.

BACKGROUND: Physicians play a fundamental role in the care of patients at the end of life that includes knowing how to accompany patients, alleviate their suffering and inform them about their situation. However, in reality, doctors are part of this society that is reticent to face death and lack the proper education to manage it in their clinical practice. The objective of this study was to explore the residents' concepts of death and related aspects, their reactions and actions in situations pertaining to death in their practice, and their perceptions about existing and necessary training conditions. METHODS: A qualitative approach was used to examine these points in depth based on interviews conducted with seven oncology residents. RESULTS: Participants do not have a clear concept of death and, although it is seen as a common phenomenon, they consider it an enemy to beat. The situations to which respondents react more frequently with frustration and sadness after the death of patients were when they felt emotionally involved, if they identify with the patient, in cases of pediatric patients and with patients who refuse treatment. To deal with death, participants raise barriers and attempt to become insensitive. Although residents in this study recognize the importance of training to learn how to better deal with death, it seems they are not fully invested in reaching more of it. CONCLUSIONS: Participants face death in a daily basis without the necessary training, which appears to impact them more than they are willing to accept. They do not achieve their goals managing situations regarding death as well as they assume they do. Despite recognizing the need of more training and support for better coping with death, they prefer to continue to learn from their experience. TRIAL REGISTRATION: Not applicable.

Cancer-associated fibroblasts predict poor outcome and promote periostin-dependent invasion in oesophageal adenocarcinoma.

Interactions between cancer cells and cancer-associated fibroblasts (CAFs) play an important role in tumour development and progression. In this study we investigated the functional role of CAFs in oesophageal adenocarcinoma (EAC). We used immunochemistry to analyse a cohort of 183 EAC patients for CAF markers related to disease mortality. We characterized CAFs and normal oesophageal fibroblasts (NOFs) using western blotting, immunofluorescence and gel contraction. Transwell assays, 3D organotypic culture and xenograft models were used to examine the effects on EAC cell function and to dissect molecular mechanisms regulating invasion. Most EACs (93%) contained CAFs with a myofibroblastic (α-SMA-positive) phenotype, which correlated significantly with poor survival [p = 0.016; HR 7. 1 (1.7-29.4)]. Primary CAFs isolated from EACs have a contractile, myofibroblastic phenotype and promote EAC cell invasion in vitro (Transwell assays, p ≤ 0.05; organotypic culture, p < 0.001) and in vivo (p ≤ 0.05). In vitro, this pro-invasive effect is modulated through the matricellular protein periostin. Periostin is secreted by CAFs and acts as a ligand for EAC cell integrins αvß3 and αvß5, promoting activation of the PI3kinase-Akt pathway. In patient samples, periostin expression at the tumour cell-stromal interface correlates with poor overall and disease-free survival. Our study highlights the importance of the tumour stroma in EAC progression. Paracrine interaction between CAF-secreted periostin and EAC-expressed integrins results in PI3 kinase-Akt activation and increased tumour cell invasion. Most EACs contain a myofibroblastic CAF-rich stroma; this may explain the aggressive, highly infiltrative nature of the disease, and suggests that stromal targeting may produce therapeutic benefit in EAC patients.

Mixed success for carbon payments and subsidies in support of forest restoration in the neotropics.

Restoration of forests in low- and middle-income countries (LMICs) has the potential to contribute to international carbon mitigation targets. However, high upfront costs and variable cashflows are obstacles for many landholders. Carbon payments have been promoted as a mechanism to incentivize restoration and economists have suggested cost-sharing by third parties to reduce financial burdens of restoration. Yet empirical evidence to support this theory, based on robust, dynamic field sampling is lacking. Here we use large, long-term datasets from Panama to evaluate the financial prospects of three forest restoration methods under different cost-sharing and carbon payment designs where income is generated through timber harvests. We show some, but not all options are economically viable. Further work combining growth and survival data from field trials with more sophisticated financial analyses is essential to understanding barriers and realizing the potential of forest restoration in LMICs to help meet global carbon mitigation commitments.

Loss of T-tubules and other changes to surface topography in ventricular myocytes from failing human and rat heart.

T-tubular invaginations of the sarcolemma of ventricular cardiomyocytes contain junctional structures functionally coupling L-type calcium channels to the sarcoplasmic reticulum calcium-release channels (the ryanodine receptors), and therefore their configuration controls the gain of calcium-induced calcium release (CICR). Studies primarily in rodent myocardium have shown the importance of T-tubular structures for calcium transient kinetics and have linked T-tubule disruption to delayed CICR. However, there is disagreement as to the nature of T-tubule changes in human heart failure. We studied isolated ventricular myocytes from patients with ischemic heart disease, idiopathic dilated cardiomyopathy, and hypertrophic obstructive cardiomyopathy and determined T-tubule structure with either the fluorescent membrane dye di-8-ANNEPs or the scanning ion conductance microscope (SICM). The SICM uses a scanning pipette to produce a topographic representation of the surface of the live cell by a non-optical method. We have also compared ventricular myocytes from a rat model of chronic heart failure after myocardial infarction. T-tubule loss, shown by both ANNEPs staining and SICM imaging, was pronounced in human myocytes from all etiologies of disease. SICM imaging showed additional changes in surface structure, with flattening and loss of Z-groove definition common to all etiologies. Rat myocytes from the chronic heart failure model also showed both T-tubule and Z-groove loss, as well as increased spark frequency and greater spark amplitude. This study confirms the loss of T-tubules as part of the phenotypic change in the failing human myocyte, but it also shows that this is part of a wider spectrum of alterations in surface morphology.

Tree diversity in a tropical agricultural-forest mosaic landscape in Honduras.

Biodiversity decline in the tropics requires the implementation of comprehensive landscape management where agricultural systems are necessarily an integral element of biodiversity conservation. This study evaluates the potential for taxonomic biodiversity conservation within an intensive livestock-agricultural-forest mosaic landscape in Catacamas, Honduras. Tree sampling was performed in 448 plots set up within different forest and agricultural land uses: secondary forests, agroforestry coffee plantations, agriculture, pastures, live fences and riparian forest. All trees with a minimum diameter at breast height of 10 cm were identified and measured. We characterized their tree structure and diversity, and compared tree diversity between the different uses. The results indicate a high degree of tree species diversity: 375 species identified, belonging to 74 families among the 15,096 trees inventoried across 84.2 hectares, including many rare species (40% of the species registered three individuals or fewer). Biodiversity indices for agroforestry coffee were found equivalent to those for natural secondary forests in the Catacamas landscape. Combining biodiversity conservation and agricultural production is possible in human-pressured tropical landscapes through tree cover maintenance. Enrichment practices combining local producers and technical knowledge may improve tree diversity in agricultural landscapes by prioritizing a mix of forest and introduced tree species (rare and with multiple uses).

Elucidation of mass transfer mechanisms in pellet formation by spheronization.

Previously published mechanisms of pellet formation during extrusion-spheronization include a transfer of material between different granules. This research aimed to specify the origin of this transfered mass, enabling further insight into the extrusion-spheronization process. Granules of various diameters were rounded simultaniously in a spheronizer to ascertain if mass transfer between smaller and larger granules is truly in balance, or if mass transfer from smaller to larger granules is preferred. Granules were also marked with a fluorescent tracer to enable quantification of mass transfer. By using differently sized and shaped granules as starting material, different modes of mass transfer were investigated. Samples were taken after various process durations to investigate the kinetics of the tranfer mechanism. It was found that both small and large granules dispense and receive mass during spheronization. In general, small granules increase their size, while large granules maintain their size or show a slight size decrease, resulting in the particularly narrow monomodal size distribution.

Single-shot phase contrast microscopy using polarisation-resolved differential phase contrast.

We present a robust, low-cost single-shot implementation of differential phase microscopy utilising a polarisation-sensitive camera to simultaneously acquire four images from which phase contrast images can be calculated. This polarisation-resolved differential phase contrast (pDPC) microscopy technique can be easily integrated with fluorescence microscopy.

Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase.

Reduced nicotinamide adenine dinucleotide (NADH) is the principal electron donor in glycolysis and oxidative metabolism and is thus recognized as a key biomarker for probing metabolic state. While the fluorescence characteristics of NADH have been investigated extensively, there are discrepancies in the published data due to diverse experimental conditions, instrumentation and microenvironmental parameters that can affect NADH fluorescence. Using a cuvette-based time-resolved spectrofluorimeter employing one-photon excitation at 375 nm, we characterized the fluorescence intensity, lifetime, spectral response, anisotropy and time-resolved anisotropy of NADH in aqueous solution under varying microenvironmental conditions, namely temperature, pH, and binding to lactate dehydrogenase (LDH). Our results demonstrate how temperature, pH, and binding partners each impact the fluorescence signature of NADH and highlight the complexity of the fluorescence data when different parameters produce competing effects. We hope that the data presented in this study will provide a reference for potential sources of variation in experiments measuring NADH fluorescence.

Application of direct stochastic optical reconstruction microscopy (dSTORM) to the histological analysis of human glomerular disease.

Electron microscopy (EM) following immunofluorescence (IF) imaging is a vital tool for the diagnosis of human glomerular diseases, but the implementation of EM is limited to specialised institutions and it is not available in many countries. Recent progress in fluorescence microscopy now enables conventional widefield fluorescence microscopes to be adapted at modest cost to provide resolution below 50 nm in biological specimens. We show that stochastically switched single-molecule localisation microscopy can be applied to clinical histological sections stained with standard IF techniques and that such super-resolved IF may provide an alternative means to resolve ultrastructure to aid the diagnosis of kidney disease where EM is not available. We have implemented the direct stochastic optical reconstruction microscopy technique with human kidney biopsy frozen sections stained with clinically approved immunofluorescent probes for the basal laminae and immunoglobulin G deposits. Using cases of membranous glomerulonephritis, thin basement membrane lesion, and lupus nephritis, we compare this approach to clinical EM images and demonstrate enhanced imaging compared to conventional IF microscopy. With minor modifications in established IF protocols of clinical frozen renal biopsies, we believe the cost-effective adaptation of conventional widefield microscopes can be widely implemented to provide super-resolved image information to aid diagnosis of human glomerular disease.

Pictures Do Influence the Decision to Transfer: Outcomes of a Telemedicine Program Serving an Eight-State Rural Population.

Triaging burn patients is a daunting task because burn injuries are rare; this inexperience leads to uncertainty in treatment and referral algorithms. Our regional burn center's catchment area includes eight states. Outlying facilities consult via telephone through the medical center's transfer center. Referring provider assessments of depth or size of injury infrequently correlates with burn provider's assessments. This causes over- and under-triage of patients managed outside of burn centers. A quality improvement telemedicine project was developed to allow burn providers to review photos with referring providers to determine best management, provide pertinent education, and initiate appropriate and timely resuscitation. Details tracked include date of service, consulting provider, follow-up education offered, and whether the image reviewed changed or confirmed the requested plan of care. Of the 155 cases between January 2017 and August 2018, 24.5% of patient images changed the initial transfer decision, and 75.5% confirmed the initial plan of care. Of the cases requiring change of plan, 60.5% were down-triaged to outpatient care and 39.5% were up-triaged to transfer. Implementation of this telemedicine program has increased efficiency of resource utilization, timely resuscitation, appropriate transfer of patients requiring admission, and real-time education. Findings suggest advanced practice providers' assessments are similar to those of referring physicians. These observations may have significant implications on Emergency Medical Treatment and Labor Act (EMTALA) guidelines defining physician to physician consultation and support efficient use of available resources. Telemedicine facilitates access to specialized care and improves fiscal responsibility.

Primary extra-gastrointestinal stromal tumor (EGIST) of the mesentery: Case report and review of literature.

INTRODUCTION: Gastrointestinal stromal tumors (GISTs) represent <1% of all gastrointestinal (GI) tumors. Extra-gastrointestinal stromal tumors (EGISTs) are mesenchymal tissue neoplasm arising outside the GI tract. This rare group comprises only 5% of all GISTs. This case demonstrates a rare entity in a patient with non-specific symptoms, a large tumor size and unremarkable past personal and family history. PRESENTATION OF CASE: We present a 45-year old man with non-specific symptoms who was diagnosed with a primary EGIST arising in the small bowel mesentery after surgery. The tumor was not compromising the GI tract and it was completely resected. The tumor was sent for pathological examination that confirmed the diagnosis. Histological examination revealed a 15 cm in diameter mass, comprised of spindle cells and high mitotic activity. Treatment with imatinib mesylate was initiated. DISCUSSION: There have been only a few previous reports of EGISTs arising from the small bowel mesentery. It is believed that EGISTs originate from cells with similar pathological characteristics and biological behaviour as the intestinal cells of Cajal. Such tumors are associated with poorer prognosis, lager tumor size and younger presentation than their GI counterparts. The preferred treatment is complete surgical resection. The addition of specific tyrosine kinase inhibitors such as imatinib mesylate is recommended for high risk patients. Even though morphological and immunohistochemical similarities between GISTs and EGISTs are described, their pathogenesis, incidence, genetic background, complications and prognosis are not completely known because they are extremely rare. CONCLUSION: EGISTs are very rare tumors which originate from cells outside the GI tract and are associated with a more aggressive biological behavior than their GI counterparts. These tumors may grow without any clinical implications and should be kept in mind in the differential diagnosis for patients presenting with an abdominal mass. Further studies are needed due to lack of large patient cohort studies and long-term follow-up regarding the prognosis and management of this rare pathology.

The molecular phenotype of human cardiac myosin associated with hypertrophic obstructive cardiomyopathy.

AIM: The aim of the study was to compare the functional and structural properties of the motor protein, myosin, and isolated myocyte contractility in heart muscle excised from hypertrophic cardiomyopathy patients by surgical myectomy with explanted failing heart and non-failing donor heart muscle. METHODS: Myosin was isolated and studied using an in vitro motility assay. The distribution of myosin light chain-1 isoforms was measured by two-dimensional electrophoresis. Myosin light chain-2 phosphorylation was measured by sodium dodecyl sulphate-polyacrylamide gel electrophoresis using Pro-Q Diamond phosphoprotein stain. RESULTS: The fraction of actin filaments moving when powered by myectomy myosin was 21% less than with donor myosin (P = 0.006), whereas the sliding speed was not different (0.310 +/- 0.034 for myectomy myosin vs. 0.305 +/- 0.019 microm/s for donor myosin in six paired experiments). Failing heart myosin showed 18% reduced motility. One myectomy myosin sample produced a consistently higher sliding speed than donor heart myosin and was identified with a disease-causing heavy chain mutation (V606M). In myectomy myosin, the level of atrial light chain-1 relative to ventricular light chain-1 was 20 +/- 5% compared with 11 +/- 5% in donor heart myosin and the level of myosin light chain-2 phosphorylation was decreased by 30-45%. Isolated cardiomyocytes showed reduced contraction amplitude (1.61 +/- 0.25 vs. 3.58 +/- 0.40%) and reduced relaxation rates compared with donor myocytes (TT(50%) = 0.32 +/- 0.09 vs. 0.17 +/- 0.02 s). CONCLUSION: Contractility in myectomy samples resembles the hypocontractile phenotype found in end-stage failing heart muscle irrespective of the primary stimulus, and this phenotype is not a direct effect of the hypertrophy-inducing mutation. The presence of a myosin heavy chain mutation causing hypertrophic cardiomyopathy can be predicted from a simple functional assay.

Automated Fluorescence Lifetime Imaging High-Content Analysis of Förster Resonance Energy Transfer between Endogenously Labeled Kinetochore Proteins in Live Budding Yeast Cells.

We describe an open-source automated multiwell plate fluorescence lifetime imaging (FLIM) methodology to read out Förster resonance energy transfer (FRET) between fluorescent proteins (FPs) labeling endogenous kinetochore proteins (KPs) in live budding yeast cells. The low copy number of many KPs and their small spatial extent present significant challenges for the quantification of donor fluorescence lifetime in the presence of significant cellular autofluorescence and photobleaching. Automated FLIM data acquisition was controlled by µManager and incorporated wide-field time-gated imaging with optical sectioning to reduce background fluorescence. For data analysis, we used custom MATLAB-based software tools to perform kinetochore foci segmentation and local cellular background subtraction and fitted the fluorescence lifetime data using the open-source FLIMfit software. We validated the methodology using endogenous KPs labeled with mTurquoise2 FP and/or yellow FP and measured the donor fluorescence lifetimes for foci comprising 32 kinetochores with KP copy numbers as low as ~2 per kinetochore under an average labeling efficiency of 50%. We observed changes of median donor lifetime ≥250 ps for KPs known to form dimers. Thus, this FLIM high-content analysis platform enables the screening of relatively low-copy-number endogenous protein-protein interactions at spatially confined macromolecular complexes.

Author Correction: Authentication and characterisation of a new oesophageal adenocarcinoma cell line: MFD-1.

A correction has been published and is appended to both the HTML and PDF versions of this paper. The error has not been fixed in the paper.

Ghrelin receptor gene polymorphisms and body size in children and adults.

BACKGROUND: The GH secretagogue receptor type 1a gene (GHSR) encodes the cognate receptor of ghrelin, a gut hormone that regulates food intake and pituitary GH secretion. Previous studies in U.S. families and a German population suggested GHSR to be a candidate quantitative locus for association with human obesity and growth. AIM: The aim of the study was to test common genetic variation in GHSR for association with body size in children and adults. METHODS: Sequencing was performed to systematically identify novel single nucleotide polymorphisms (SNPs) in GHSR. A set of three haplotype-tagging SNPs that captured all the genetic variation in GHSR was identified. These three haplotype-tagging SNPs were then genotyped in three large population-based U.K. cohort studies (two adult and one childhood cohort) comprising 5807 adults and 843 children. RESULTS: No significant genotype or haplotype associations were found with adult or childhood height, weight, or body mass index. CONCLUSION: Common variation in GHSR is not associated with body size in U.K. adults or children.

Salud bucal en la primera infancia: estrategia con agentes educativas y acudientes / Oral health in early childhood: strategy with educational agents and guardians

Introducción: la dentición temporal es esencial para la nutrición, habla y autoestima. Los niños y niñas menores de 5 años no poseen habilidades motoras para efectuar higiene bucal por sí mismos. Objetivo: diseñar e implementar una estrategia educativa para la promoción de la salud bucal en la primera infancia con agentes educativas y acudientes de los niños y niñas que asisten a los hogares comunitarios del Instituto Colombiano de Bienestar Familiar en Santa Rosa de Cabal (Colombia) en el 2019. Materiales y métodos: investigación cualitativa, diseño investigación-acción. Muestreo no probabilístico por conveniencia, participaron 25 agentes educativas y 55 acudientes. Técnicas de investigación empleadas: grupos focales y talleres pedagógicos. Para analizar los datos se usó Análisis Temático. Resultados: se desarrolló en 3 fases. 1) los/as participantes expresaron que tener salud bucal consiste en la adecuada higiene, aunque todos/as conocían la caries, los cuidadores no la comprendían como una patología, las agentes educativas percibieron tener alta capacidad de autoeficacia para contribuir a la salud bucal de los niños/as. 2) En los talleres pedagógicos se resolvieron inquietudes en temas como crecimiento y desarrollo del sistema estomatognático, enfermedades bucales, entre otros. 3) Las agentes educativas aprendieron la relación de la salud bucal con lactancia materna, asimismo las causas y consecuencias de las patologías bucales, y replicaron lo aprendido. Conclusiones: se recomienda el desarrollo de estrategias educativas guiadas por modelos teóricos y con metodologías participativas, adaptadas a las necesidades y contextos de las comunidades, pues se requiere producir conocimiento situado y con aceptabilidad cultural.

Introduction: Primary dentition is essential for nutrition, speech, and self-esteem. Children under the age of 5 do not have the motor skills to perform oral hygiene on their own. Objective: To design and implement an educational strategy to promote oral health in early childhood with educators and parents/guardians of children attending the community homes of the Colombian Family Welfare Institute in Santa Rosa de Cabal (Colombia) in 2019. Materials and methods: Qualitative, action research design. Non-probability convenience sampling was used. Twenty-five educators and 55 parents/guardians participated. Focus groups and pedagogical workshops were the research techniques used. Thematic analysis was performed for data analysis. Results: It was conducted in three phases: 1) Participants expressed that oral health is about proper hygiene. Although all were aware of caries, the caregivers did not understand it as a pathology. The educators perceived that they had high self-efficacy to contribute to children's oral health; 2) in the pedagogical workshops, concerns were resolved on topics such as growth and development of the stomatognathic system, oral diseases, among others, and 3) the educators learned about the relationship between oral health and breastfeeding, as well as the causes and consequences of oral pathologies, and reproduced what they learned. Conclusions: The development of educational strategies based on theoretical models and participatory methods adapted to the needs and contexts of the communities is recommended since it is necessary to produce situated knowledge with cultural acceptance.

Introdução: a dentição temporária é essencial para nutrição, fala e autoestima. Meninos e meninas menores de 5 anos não possuem habilidades motoras para realizar a higiene bucal sozinhos. Objetivo: desenhar e implementar uma estratégia educativa para a promoção da saúde bucal na primeira infância com agentes educativos e responsáveis dos meninos e meninas que frequentam as casas comunitárias do Instituto Colombiano de Bem-Estar Familiar em Santa Rosa de Cabal (Colômbia) em 2019. Materiais e Métodos: pesquisa qualitativa, desenho de pesquisa-ação. Amostragem não probabilística por conveniência, participaram 25 agentes educativos e 55 responsáveis. Técnicas de pesquisa utilizadas: grupos focais e seminários educativos. Para analisar os dados se usou Análises Temático. Resultados: foi desenvolvido em 3 fases. 1) os participantes expressaram que ter saúde bucal consiste em higiene adequada, embora todos soubessem sobre a cárie, os cuidadores não a entendiam como uma patologia, os agentes educativos percebiam que tinham alta capacidade de autoeficácia para contribuir com a saúde bucal das crianças 2) Nos seminários educativos foram sanadas dúvidas sobre temas como crescimento e desenvolvimento do sistema estomatognático, doenças bucais, entre outros. 3) Os agentes educativos aprenderam a relação entre saúde oral e amamentação, bem como as causas e consequências das patologias orais, e replicaram o que aprenderam. Conclusões: Recomenda-se o desenvolvimento de estratégias educativas pautadas em modelos teóricos e com metodologias participativas, adaptadas às necessidades e contextos das comunidades, pois é necessário produzir conhecimento situado com aceitabilidade cultural.

Endometriosis de la pared abdominal en una paciente con antecedente de histerectomía. Reporte de caso / Abdominal wall endometriosis in a patient with a history of hysterectomy. Case report

Resumen ANTECEDENTES: La endometriosis de la pared abdominal implica la coexistencia de tejido endometrial en la superficie peritoneal parietal; la incidencia reportada es de 0.03 a 3.5%. Su causa aún no está debidamente esclarecida. CASO CLÍNICO: Paciente de 35 años, con antecedentes ginecoobstétricos de: tres embarazos, tres cesáreas, última cinco años previos a la intervención, en la que se practicó una histerectomía obstétrica indicada por sangrado transoperatorio. El padecimiento actual se inició 24 horas previas a su ingreso a Urgencias, con dolor espontáneo en la fosa iliaca derecha, de difícil relación con los ciclos menstruales debido al antecedente quirúrgico, acompañado de aumento de volumen y náuseas. En la exploración inicial se identificaron dos masas intraabdominales que se confirmaron en la tomografía computada, situadas por encima de la aponeurosis. Se procedió a la intervención quirúrgica para extirpación de ambas masas. El estudio histopatológico reportó: tumores compatibles con endometriosis. CONCLUSIÓN: La endometriosis es un padecimiento con alta prevalencia en el mundo, no así en su ubicación en la pared abdominal. A pesar de que aún no se conoce con certeza su causa, se sabe que la inoculación directa (muchas veces debida a un procedimiento ginecológico quirúrgico) y la proliferación celular tienen participación relevante en su origen.

Abstract BACKGROUND: Abdominal wall endometriosis is the coexistence of endometrial tissue on the parietal peritoneal surface with a reported incidence of 0.03 to 3.5%. Its cause is not well understood. CLINICAL CASE: 35-year-old female patient with a gyneco-obstetric history of: three pregnancies, three cesarean sections, last five years prior to surgery, in which an obstetric hysterectomy was performed, indicated by transoperative bleeding. The current presentation began 24 hours before her admission to the emergency department with spontaneous pain in the right iliac fossa, difficult to relate to menstrual cycles due to her surgical history, accompanied by increased volume and nausea. Initial examination revealed two intra-abdominal masses, confirmed by computed tomography, located above the aponeurosis. Surgery was performed to remove both masses. Histopathologic examination revealed tumors compatible with endometriosis. CONCLUSION: Endometriosis is a very common disease in the world, but not in the abdominal wall. Although its cause is still not known with certainty, it is known that direct inoculation (often due to gynecologic surgery) and cell proliferation play a relevant role in its origin.

Examining the candidacy of ghrelin as a gene responsible for variation in adult stature in a United Kingdom population with type 2 diabetes.

CONTEXT: Recently, a quantitative trait locus for stature was reported on chromosome 3p26 in patients with type 2 diabetes. OBJECTIVE: Given that ghrelin is a peptide involved in GH release and located on 3p26, we hypothesized that variation within its gene (GHRL) may be responsible for the quantitative trait locus on 3p26. DESIGN: The evidence for linkage around GHRL was refined with the genotyping of an additional four microsatellites (D3S4545, D3S1537, D3S1597, and D3S3611), giving a total of 27 markers, followed by multipoint variance components linkage analysis. Probands from the linkage families were typed for five common single nucleotide polymorphisms (SNPs) within GHRL and tested for association with adult stature using haplotype trend regression. RESULTS: The maximum multipoint evidence for linkage between adult stature and the 27 microsatellites yielded an LOD score of 2.58 (P = 0.0003) between D3S1297 and D3S1304. Five common (frequency of > or =5%) SNPs were typed in the probands [two promoter SNPs (rs27647 and rs26802), two exonic (rs696217 and rs4684677), and one intronic (rs35683)] capturing 80% of the total common variation in GHRL. No association was found between any SNP (or haplotypes thereof) and adult stature. CONCLUSION: Common genetic variation within GHRL is not responsible for variation in adult stature in this population.

Ghrelin and cardiovascular health.

Ghrelin and its receptor are widely distributed in cardiovascular tissues, and there is no doubt that the effects of ghrelin in the cardiovascular system are mediated not only via its growth-hormone-releasing effect but also by direct effects on the heart. Indeed, new pharmacological approaches with animal and cell models using elegant study designs have described new functions of ghrelin, providing new potential therapeutic opportunities for ghrelin in cardiovascular medicine.