Reciprocal regulation between the molecular clock and kidney injury.

Tubulointerstitial fibrosis is the common pathological substrate for many etiologies leading to chronic kidney disease. Although perturbations in the circadian rhythm have been associated with renal disease, the role of the molecular clock in the pathogenesis of fibrosis remains incompletely understood. We investigated the relationship between the molecular clock and renal damage in experimental models of injury and fibrosis (unilateral ureteral obstruction, folic acid, and adenine nephrotoxicity), using genetically modified mice with selective deficiencies of the clock components Bmal1, Clock, and Cry We found that the molecular clock pathway was enriched in damaged tubular epithelial cells with marked metabolic alterations. In human tubular epithelial cells, TGFß significantly altered the expression of clock components. Although Clock played a role in the macrophage-mediated inflammatory response, the combined absence of Cry1 and Cry2 was critical for the recruitment of neutrophils, correlating with a worsening of fibrosis and with a major shift in the expression of metabolism-related genes. These results support that renal damage disrupts the kidney peripheral molecular clock, which in turn promotes metabolic derangement linked to inflammatory and fibrotic responses.

Melanoma RBPome identification reveals PDIA6 as an unconventional RNA-binding protein involved in metastasis.

RNA-binding proteins (RBPs) have been relatively overlooked in cancer research despite their contribution to virtually every cancer hallmark. Here, we use RNA interactome capture (RIC) to characterize the melanoma RBPome and uncover novel RBPs involved in melanoma progression. Comparison of RIC profiles of a non-tumoral versus a metastatic cell line revealed prevalent changes in RNA-binding capacities that were not associated with changes in RBP levels. Extensive functional validation of a selected group of 24 RBPs using five different in vitro assays unveiled unanticipated roles of RBPs in melanoma malignancy. As proof-of-principle we focused on PDIA6, an ER-lumen chaperone that displayed a novel RNA-binding activity. We show that PDIA6 is involved in metastatic progression, map its RNA-binding domain, and find that RNA binding is required for PDIA6 tumorigenic properties. These results exemplify how RIC technologies can be harnessed to uncover novel vulnerabilities of cancer cells.

Evaluation of structural and ultrastructural changes in thyroid and parathyroid glands after near infrared irradiation: study on an animal model.

Intraoperative identification of parathyroid glands is a tough task for surgeons performing thyroid or parathyroid surgery, because the small size, color and shape of these glands hinder their discrimination from other cervical tissues. In 2011, Paras described the autofluorescence of parathyroid glands, a property that could facilitate their intraoperative identification. Parathyroid glands submitted to a 785 nm laser beam emit fluorescence in the near infrared range, with a peak at 822 nm. As the intrinsic properties of secretory tissues may be affected by the exposure to the near infrared light, a situation that could preclude their intraoperative utilization, the authors compared the structural and ultra-structural patterns of rat's thyroid and parathyroid glands submitted to irradiation replicating the conditions that allow their intraoperative identification, with those of non irradiated animals. Twenty-four Wistar rats were divided into six groups: animals of Groups 1, 3 and 5 were submitted under general anesthesia to direct irradiation of the cervical area with a 780 nm LED light for 3 minutes through a cervical incision, and animals of Groups 2, 4 and 6 were submitted to cervical dissection without irradiation. Animals of were euthanized immediately (Groups 1 and 2), at Day 30 (Groups 3 and 4) at and at Day 60 (Groups 5 and 6) and thyroid and parathyroid glands were removed: one lobe was prepared for conventional pathological examination and the other lobe for electron microscopy observed by three experienced pathological experts. Twenty-four samples were prepared for conventional histology and there were no alterations reported in any group. Due to technical problems, only 21 samples were observed by electron microscopy and there were no differences in the ultrastructure of parathyroid and thyroid glands, namely the nuclear pattern, mitochondria, endoplasmic reticulum or secretory granules, in any of the groups. These results confirm the innocuity of near infrared irradiation', allowing its intraoperative utilization.

Spectroscopic Analysis of Parathyroid and Thyroid Tissues by Ground-State diffuse Reflectance and Laser Induced Luminescence: a Preliminary Report.

Intraoperative discrimination of thyroid and parathyroid tissues is fundamental in thyroid surgery. Recent fluorescence studies have shown stronger NIR emission in parathyroid tissue than in thyroid tissue, presenting a potential avenue for the development of a tool for surgical assistance. However, the fluorophore responsible for this emission has not yet been identified. In this work, spectroscopic analysis was performed to ascertain the origin of the emission peaks in parathyroid tissue. Ground-state diffuse reflectance (GSDR) absorption spectroscopy and laser-induced luminescence (LIL) emission spectroscopy were performed in parathyroid, thyroid, and fatty tissue samples and the resulting spectra were compared with the peaks of known fluorophores to identify the origin of each peak. The spectra of the different tissue types were also compared in order to evaluate the wavelength which presents the highest parathyroid emission relative to the emission of the surrounding tissues, representing the ideal wavelength for parathyroid detection. An emission peak in these conditions was observed for both thyroid and parathyroid tissue at 711 nm, with a higher intensity in parathyroid sample, making it suitable for detection applications. These results show a potential avenue for the development of a system allowing parathyroid detection in a surgical setting.

Identification of inadvertently removed parathyroid glands during thyroid surgery using autofluorescence.

BACKGROUND: Incidental excision of parathyroid glands is a common event during thyroid surgery and in spite the divergent results that can be obtained from the literature about its clinical significance, all efforts must be used to their preservation. Due to the autofluorescence emitted by parathyroid glands, authors began to use a custom device to inspect thyroidectomy specimens for incidentally removed parathyroid tissue; the results of using this device are presented in this manuscript. METHODS: Specimens of 40 consecutive thyroid surgeries were inspected. Localization of suspect high-fluorescence spots were recorded for confirmation with a pathological exam. Determinations of calcium and parathyroid hormone (PTH) were completed prior to surgery and at 24 hours and 15 days after the operation. RESULTS: Patient age ranged from 36 to 83 years and were predominantly female (82.5%). Calcium values at 24 hours post-operation varied between 7.1 and 9.5. The PTH values ranged between 3 and 77. Thirteen patients (32.5%) presented with biochemical hypocalcemia at 24 hours. At 15 days after the operation, only one patient presented with a calcium value below 8 (PTH: 10.9) with complete normalization 6 months after the surgery. Pathological examination identified eight parathyroid fragments in seven patients. There was no correlation between the presence of parathyroid tissue in the specimen and post-operative hypocalcemia (P=0.254). Eight suspicious areas of augmented fluorescence where detected; seven were coincident with the pathological examination and one was a false positive. One intrathyroidal gland was not identified, resulting a sensitivity of 87.5% and specificity 96.2%. CONCLUSIONS: Although no correlation between incidental parathyroidectomy and hypocalcemia was demonstrated, autofluorescence may be a useful tool for in-table identification of incidentally-removed glands.

Fibrous solitary tumor, a rare and ubiquitous neoplasy.

Fibrous solitary tumors (FST) are mesenchymal tumors that can appear in different body regions. It is estimated that around 30% are found in the thoracic region, while rarely in meninges, abdomen, pelvis, extremities and bones. A correct diagnosis is important because 15-20% of cases develop a malignant behavior. Treatment of choice is surgical and posterior follow-up is essential. We present two atypical extrapleural FST cases, diagnosed in our center. Both were treated with surgery and in one case arterial embolization to reduce the bleeding risk was previously done.

Near-infrared irradiation of the thyroid area: effects on weight development and thyroid and parathyroid secretory patterns.

This study evaluates possible changes in weight and the secretory patterns of the thyroid and parathyroid glands irradiated with a 780-nm light-emitting diode (LED) source under conditions allowing their identification in an animal model. The use of near-infrared light sources to detect the parathyroid glands has been a subject of research due to the benefit provided to patients undergoing thyroid and parathyroid surgery. However, few studies have explored possible changes in weight and the secretory patterns of the glands when subjected to light stimulation. Thirty-two male Wistar rats were divided into two groups and subjected to cervical dissection and irradiation of the thyroid-parathyroid region under general anesthesia. The animals in group 1 (irradiated) were exposed to a 780-nm LED light beam for 180 s (dose 1.37 J/cm2), whereas the animals in group 2 (control) were not irradiated. Blood samples were collected pre-exposure, 7 min after exposure, and 30 and 60 days after exposure to measure calcium, parathyroid hormone (PTH), triiodothyronine (T3), thyroxine (T4), and thyroid-stimulating hormone (TSH) levels in both groups. Weight variations between the evaluation periods were also analyzed. Parametric statistics were used (Student's t test and ANOVA) after assuming normal distribution of continuous variables with the Shapiro-Wilk test. No significant variation was observed in any of the analyzed parameters pre- and postexposure. A significant increase in the calcium level was observed at 30 days in the irradiated group compared with that in the control group (11.156 ± 1.31 mg/dl vs 10.300 ± 0.30 mg/dl; df = 30 p < 0.03) but this change was no longer observed at 60 days (9.925 ± 0.23 mg/dl vs 10.014 ± 0.18 mg/dl; df = 14 p = 0.21). Irradiated rats gained less weight in the first 30 days after surgery (39.647 ± 32.39 vs 55.917 ± 22.61; df = 30 p = 0.146) and at 60 days (84.000 ± 27.36 vs 84.571 ± 5.62; df = 14 p = 0.957), no differences were observed between the two groups. No significant variations in weight development or changes in the secretory patterns of the thyroid and parathyroid glands exposed to near-infrared stimulation were observed, confirming the safety of this light source in the identification of the parathyroid glands.

[Twenty-five years of duodenal switch. How to switch to the duodenal switch]. / Veinticinco años de cruce duodenal. Cómo cambiar al cruce.

INTRODUCTION: Background: the duodenal switch (DS) is a procedure that combines a vertical gastrectomy (VG) plus a biliopancreatic diversion (BPD). Objectives: to report our experience in 950 consecutive DS patients with morbid obesity (MO) performed from 1994 to 2011, with 27 years of follow-up. Environment: mix of teaching and private institution in a regional hospital in Spain. Methods: retrospective review of 950 consecutive morbidly obese patients treated with DS surgery. Results: five hundred and eighteen open DS (ODS) and 432 laparoscopic DS (LDS) were performed. Operative mortality was 0.84% (1.38% in ODS and 0.38% in LDS); 4.84% had one leak, two had liver failure (0.2%) and malnutrition was present in 3.1%. At five years, the body mass index (BMI) percentage of lost overweight (%EWL) was 80% and the percentage of expected BMI loss was more than 100%. Conclusions: the DS is the most aggressive bariatric technique but with the best long-term weight loss. Operative complications and long-term follow-up guidelines are described.

INTRODUCCIÓN: Antecedentes: el cruce duodenal (CD) es un procedimiento que combina una gastrectomía vertical (GV) más una derivación biliopancreática (DBP). Objetivos: informar de nuestra experiencia en 950 CD consecutivos en pacientes con obesidad mórbida (OM) realizados de 1994 a 2011 y con 27 años de seguimiento. Entorno: mezcla de enseñanza e institución privada en un hospital comarcal de España. Métodos: revisión retrospectiva de 950 pacientes consecutivos con obesidad mórbida tratados con cirugía de CD. Resultados: se realizaron 518 CD abiertos (CDA) y 432 CD laparoscópicos (CDL). La mortalidad operatoria fue del 0,84% (1,38% en CDA y 0,38% en CDL). El 4,84% tuvo una fuga, dos tuvieron insuficiencia hepática (0,2%) y la desnutrición estuvo presente en el 3,1%. A los cinco años, el porcentaje de sobrepeso perdido (PSP) de índice de masa corporal (IMC) fue del 80% y el porcentaje de pérdida esperada de IMC fue más del 100%. Conclusiones: el CD es la técnica bariátrica más agresiva pero con mejor pérdida de peso a largo plazo. Se describen las complicaciones operatorias y pautas de seguimiento a largo plazo.

Parathyroid identification by autofluorescence - preliminary report on five cases of surgery for primary hyperparathyroidism.

BACKGROUND: Intra-operative identification of parathyroid glands is often a challenge for surgeons performing parathyroid or thyroid surgery. Parathyroid glands stimulated by near-infrared light emit autofluorescence, which allows their discrimination from all other tissues in the region, and this may be of value during thyroid and parathyroid surgery. In this study, we present the results of the utilization of a low-cost device developed for the identification of parathyroid glands in surgery for primary hyperparathyroidism. CASE PRESENTATION: In 5 patients operated in our hospital with the diagnosis of primary hyperparathyroidism and non-concordant ultrasonography and Sestamibi scan, we used a 780 nm Light Emitting Diode (LED) to stimulate the cervical area. The resulting autofluorescence was visualized with night vision goggles with a 832 nm filter assembled. In all the five patients, an easily distinguishable nodule was identified and excised, and confirmed as parathyroid adenoma by histological exam. Intra-operative PTH assay showed significant decrease compared with basal values, fulfilling the Miami Criteria for surgical success in use in our institution. CONCLUSION: The utilization of autofluorescence for intra-operative identification of parathyroid glands may have a clinical application in surgery for primary hyperparathyroidism, being of special utility when ultrasonography and Sestamibi Scan are non concordant.

Multiple facial plaque variant of trichoblastoma.

BACKGROUND: The plaque variant of trichoblastoma has been described as a solitary tumor with diffuse infiltration of the lower dermis and hypodermis, with poorly defined borders. Herein, we report a new variant of multiple centrofacial trichoblastoma. OBJECT: To describe clinical and pathological features of a new multiple kind of plaque variant of centrofacial trichoblastoma. METHODS: Case series of patients with a multiple-plaque variant of centrofacial trichoblastoma treated in our department between 2005 and 2017. We identified eight patients with the centrofacial plaque variant of trichoblastoma treated in our department from 2005 to 2017. RESULTS: The final study sample comprised 13 trichoblastomas from four patients. All patients also developed at least one basal cell carcinoma. Mohs surgery was the method of treatment in the majority of the cases of trichoblastoma and in all the cases of basal cell carcinoma. We needed between 2 and 6 stages to obtain free margins in our cases of facial plaque trichoblastomas treated by Mohs surgery. CONCLUSION: To the best of our knowledge, a multiple-plaque variant of trichoblastoma has not been described in the literature. We suggest a genetic origin of this variant of trichoblastoma and describe its remarkable infiltrative nature, with poorly defined surgical margins.

Four decades of bariatric surgery in a community hospital of Spain.

INTRODUCTION: Bariatric surgical practice changes in the community setting may be under-reported. We present the developments in a Spanish bariatric surgical practice in the community setting of Alcoy from its origin in 1977 through the present. METHODS: Bariatric surgical techniques employed in a country community setting over the course of nearly four decades were reviewed retrospectively and qualitatively. RESULTS: Surgeons and medical professionals from Alcoy, Spain were involved in the evolution of bariatric surgery patient management and surgical technique from 1977s through 2017. During the last 40 years, 1,475 patients were treated in our clinics. Spanish bariatric surgeons contributed to advances in gastric bypass in the 1970s, vertical banded gastroplasty in the 1980s, bilio-pancreatic diversion/duodenal switch in the 1990s, and innovations associated with laparoscopy from the 1990s onward. Outcomes and approaches to prevention and treatment of bariatric surgical complications are reviewed from a community perspective. Contributions to the bariatric surgical nomenclature and weight-loss reporting are noted. CONCLUSIONS: The practice of bariatric surgery in the community setting must be updated continuously, as in any human and surgical endeavor. Medical professionals in community bariatric practices should contribute their experiences to the field through all avenues of scientific interaction and publication.

Identification of non-coding genetic variants in samples from hypoxemic respiratory disease patients that affect the transcriptional response to hypoxia.

A wide range of diseases course with an unbalance between the consumption of oxygen by tissues and its supply. This situation triggers a transcriptional response, mediated by the hypoxia inducible factors (HIFs), that aims to restore oxygen homeostasis. Little is known about the inter-individual variation in this response and its role in the progression of disease. Herein, we sought to identify common genetic variants mapping to hypoxia response elements (HREs) and characterize their effect on transcription. To this end, we constructed a list of genome-wide HIF-binding regions from publicly available experimental datasets and studied the genetic variability in these regions by targeted re-sequencing of genomic samples from 96 chronic obstructive pulmonary disease and 144 obstructive sleep apnea patients. This study identified 14 frequent variants disrupting potential HREs. The analysis of the genomic regions containing these variants by means of reporter assays revealed that variants rs1009329, rs6593210 and rs150921338 impaired the transcriptional response to hypoxia. Finally, using genome editing we confirmed the functional role of rs6593210 in the transcriptional regulation of EGFR. In summary, we found that inter-individual variability in non-coding regions affect the response to hypoxia and could potentially impact on the progression of pulmonary diseases.

Stapling the bougie in sleeve gastrectomy: video.

Laparoscopic sleeve gastrectomy has become the most popular bariatric surgical operation. Based on the lesser curvature, the sleeve is created with endo-staplers, which separate the lesser curvature longitudinally and vertically from the rest of the stomach, including the greater curvature. The use of a bougie (a medical instrument used in an esophageal dilation) helps to calibrate the diameter of the sleeve. Intraoperative bougie stapling is a serious complication that should be prevented; when it happens, correction is mandatory. We present 3 cases with a video.

Laparoscopic gastric sleeve, subtotal antrectomy and omentoplasty.

Sleeve-forming gastrectomy (SFG) is the operation to make a gastric sleeve (GS). The video presents the subtotal removal of the antrum and the use of sliding self-locking stitch and Aberdeen knots as suture reinforcement with omentoplasty of the GS staple line with the aim of decreasing bleeding and leaks.

Fluorescence in situ hybridization for the differential diagnosis between Spitz naevus and spitzoid melanoma.

AIMS: The differential diagnosis between Spitz naevus and spitzoid melanoma can be extremely difficult, or even impossible. In recent years, many attempts have been made to find specific histopathological or immunohistochemical markers, although none has proved successful. Because the prognosis and treatment of each are very different, it is important to distinguish between these entities. We evaluated the ability of the fluorescence in situ hybridization (FISH) assay-designed to detect the copy number of the RREB1 (6p25), MYB (6q23) and CCND1 (11q13) genes and of centromere 6 (Cep 6)-in order to distinguish between Spitz naevus and spitzoid melanoma. METHODS AND RESULTS: We evaluated 12 spitzoid melanomas and six Spitz naevi from our records. The diagnosis of both conditions was based on previously described histopathological criteria. We obtained valuable results for FISH in eight spitzoid melanomas and five Spitz naevi. Chromosomal aberrations were detected in seven of the eight spitzoid melanomas (FISH-positive) and in none of the five Spitz naevi. The FISH-negative spitzoid melanoma was the least typical in its group. CONCLUSIONS: FISH was able to distinguish between Spitz naevus and spitzoid melanoma, with a sensitivity of 87.5% and a specificity of 100%. Our findings suggest that FISH could prove a useful tool in the differential diagnosis between these entities.

Weight loss reporting: predicted body mass index after bariatric surgery.

BMI and %EBMIL are the most accurate methods for comparing results of patients after bariatric surgery. %EBMIL is based on BMI 25 as a constant end-point for all patients, but BMI 25 is easily achieved by patients with BMI < 50, whereas it is not so feasible for patients with BMI > 50. We were prompted to obtain by statistical methods a mathematical formula able to calculate the final BMI (FBMI) 3 years after the operation, dependent on the initial or preoperative BMI (IBMI) of a multicenter group of morbid obese patients operated with different bariatric techniques. We also obtained a specific formula for each bariatric procedure of this group of patients. We propose the name Predicted BMI for the value obtained with these formulas and its application in the %EBMIL instead of the constant value of BMI 25. We have analyzed the IBMI and FBMI of a multicenter group of 7,410 patients, subjected to different bariatric procedures with a minimum follow-up of 36 months. Statistical methods with a linear regression model have been used to obtain the two types (global and specific) of Predicted BMI. We first obtained a general formula of PBMI = IBMI x 0.4 + 11.75 for the total population of patients, and a second specific formula for each bariatric technique: PBMI = IBMI x 0.43 + 13.25 + technique_correction_adjustment. Predicted BMI and its application to the %EBMIL may result in a more rational comparison of results of bariatric patients, bariatric techniques, and groups of bariatric surgeons. Predicted BMI may advance the BMI that each patient would probably achieve after surgery.

Planejamento local, Saúde Ambiental e Estratégia Saúde da Família: uma análise do uso de ferramentas de gestão para a redução do risco de contaminação por enteroparasitoses no município de Venda Nova do Imigrante / Local planning, Environmental Health and Family Health Strategy: an analysis of the use of management tools to reduce the risk of enteroparasitosis contamination in the city of Venda Nova do Imigrante

Este estudo buscou entender como uma equipe de Saúde da Família (EqSF) identifica as problemáticas ambientais presentes em seu território de abrangência, que potencializam os riscos de contaminação por enteroparasitoses, no município de Venda Nova do Imigrante, Estado do Espírito Santo. Analisa ainda as ações de vigilância ambiental e a utilização dos dados da ficha A do Sistema de Informação da Atenção Básica (SIAB) relacionados ao meio ambiente para o planejamento, visando à redução da prevalência dessas doenças. Optou-se por uma abordagem qualitativa - estudo de caso - com finalidade descritiva, e pela técnica de grupo focal para a coleta de dados. A análise das categorias selecionadas mostrou que o entendimento dos profissionais de saúde em relação ao conceito de meio ambiente ainda se restringe a uma visão naturalista - que não relaciona o ser humano à dinâmica ecológica da vida de um ecossistema - e que a EqSF não utiliza os dados fornecidos pela ficha A para o planejamento de ações relacionadas às questões ambientais. Quanto à interação com os demais setores governamentais, não-governamentais e a própria população, encontramos inércia no que se refere ao meio ambiente, pois não há diálogo nem empoderamento dessas questões, pela equipe de saúde da família ou pela população.

This study sought to understand how a Family Health team (FHT) identifies the environmental issues present in their geographic jurisdiction, which potentiate the risk of contamination with intestinal parasites in the city of Venda Nova do Imigrante, Espírito Santo state, Brazil. It also examines the actions of environmental monitoring and the use of data from the A form of the Primary Care Information System (SIAB) related to environmental planning, aiming at reducing the prevalence of such diseases. We chose a qualitative approach - case study - with descriptive purpose, and the focus group technique for data collection. Analysis of selected categories showed that the understanding of health professionals about the concept of environment is still confined to a naturalistic view - which does not relate the human being to the ecological dynamics of the life of an ecosystem - and that the FHT does not use data provided by the A form to plan actions related to environmental issues. As for the interaction with other government sectors, non-governmental organizations and the population itself, we found inertia concerning environment, since there is no dialogue or empowerment of these issues on the part of the FHT or the population.