A new systematization of histological analysis for the diagnosis of Hirschsprung's disease

Abstract Background: Hirschsprung's Disease (HD) is characterized by intestinal sub-occlusion and the absence of enteric ganglion cells. A rectal biopsy examination is performed to confirm the diagnosis. In a recent study, we demonstrated that the analysis of 60 sections of rectal mucosa and submucosa stained by H&E may ensure a 90% diagnostic accuracy. Although the need to analyze so many sections makes the process of reading the slides more time-consuming, this encouraged us to study their distribution in the healthy rectal submucosa, to simplify the diagnosis. Objectives: To develop a method that facilitates HD diagnosis by studying the distribution of ganglion cells in the submucosal plexus. Methods: Using the calretinin technique, we studied the distribution of plexuses in 60 fragments of rectal submucosa from 19 cadavers. After the study, the reading method created was used for diagnosis in 47 cases of suspected HD, using H&E staining. The accuracy was verified by comparing the results obtained with H&E to those obtained with the acetylcholinesterase technique, the golden standard in our laboratory. Results: The study of submucosal plexus distribution showed that just by examining the submucosal region every 20 µm, approximately, it is possible to locate a ganglionic plexus, and we have already been able to diagnose HD with 93% accuracy. Conclusion: The study of ganglion cell distribution enabled the creation of a simplified method for reading the slides. The method applied achieved good accuracy and it can be used as an alternative method in HD diagnosis.

Effects of local and remote ischemic postconditioning methods on ischemiareperfusion injury in a young animal model of acute mesenteric ischemia

Purpose: Acute mesenteric ischemia (AMI) is a condition in pediatric surgery that ranges from intestine necrosis to death. Ischemic postconditioning (IPoC) methods were developed to reduce the damage caused by revascularization. This study aimed to evaluate the efficacy of these methods in an experimental weaning rat model. Methods: Thirty-two 21-day-old Wistar rats were allocated into four groups according to the surgical procedure performed: control, ischemia-reperfusion injury (IRI), local (LIPoC) and remote IPoC (RIPoC). At euthanasia, fragments of the intestine, liver, lungs, and kidneys were submitted to histological, histomorphometric, and molecular analyses. Results: In the duodenum, intestines, and kidneys histological alterations promoted by IRI were reversed by remote postconditioning method. In the distal ileum, the histomorphometric alterations could be reversed by the postconditioning methods with more evident effects promoted by the remote method. The molecular analysis found that the levels of expression of Bax (proapoptotic) and Bcl-XL (antiapoptotic) genes in the intestine were increased by IRI. These alterations were equally reversed by the postconditioning methods, with more evident effects of the remote method. Conclusions: IPoC methods positively reduced the damage caused by IRI in weaning rats.

Association between psychosocial factors in workers and multisite pain: cross-sectional study / Associação entre fatores psicossociais em trabalhadores e dor multirregional: estudo transversal

ABSTRACT BACKGROUND AND OBJECTIVES: Psychosocial factors may be associated with multisite pain, which is characterized by pain symptoms in more than one part of the body. The aim of the present study was to determine associations between psychosocial factors and multisite pain in a population of workers. METHODS: A cross-sectional study was conducted involving 195 workers (educators, administrative technicians, healthcare workers, cleaners, and zookeepers). Psychosocial factors were evaluated using the short form of the second version of the Copenhagen Psychosocial Questionnaire. Multisite pain was identified using the Nordic Musculoskeletal Questionnaire. RESULTS: Multisite pain was associated with quantitative demands (OR=1.31; 95% CI: 1.06-1.63), work pace (OR=1.20; 95% CI: 1.01-1.43), emotional demands (OR=1.39; 95% CI: 1.18-1.63), commitment to the workplace (OR=0.75; 95% CI: 0.62-0.91), predictability (OR=0.86; 95% CI: 0.76-0.99), job satisfaction (OR=0.53; 95% CI: 0.32-0.88), work-family conflict (OR=1.37; 95% CI: 1.16-1.62), justice (OR=0.81; 95% CI: 0.69-0.94), general health perception (OR=0.54; 95% CI: 0.38-0.76), burnout (OR=1.41; 95% CI: 1.17-1.69), and stress (OR=1.41; 95% CI: 1.18-1.68). CONCLUSION: Several psychosocial factors were associated with multisite pain, indicating that these factors could be considered in the multisite pain management.

RESUMO JUSTIFICATIVA E OBJETIVOS: Fatores psicossociais podem estar associados à dor multirregional, caracterizada por sintomas de dor em mais de uma parte do corpo. O objetivo do presente estudo foi determinar associações entre fatores psicossociais e dor multirregional em uma população de trabalhadores. MÉTODOS: Foi realizado um estudo transversal com 195 trabalhadores (professores, técnicos administrativos, profissionais de saúde, faxineiros e funcionários de zoológicos). Os fatores psicossociais foram avaliados por meio da versão abreviada da segunda versão do Copenhagen Psychosocial Questionnaire. A dor multirregional foi identificada por meio do Nordic Musculoskeletal Questionnaire. RESULTADOS: A dor multirregional foi associada com demandas quantitativas (OR=1,31; IC 95%: 1,06-1,63), ritmo de trabalho (OR=1,20; IC 95%: 1,01-1,43), demandas emocionais (OR=1,39; IC 95%: 1,18- 1,63), compromisso com o local de trabalho (OR=0,75; IC 95%: 0,62-0,91), previsibilidade (OR=0,86; IC 95%: 0,76-0,99), satisfação no trabalho (OR=0,53; IC 95%: 0,32-0,88), conflito trabalho-família (OR=1,37; IC 95%: 1,16-1,62), justiça (OR=0,81; IC 95%: 0,69-0,94), percepção geral de saúde (OR=0,54; IC 95%: 0,38-0,76), burnout (OR=1,41; IC 95%: 1,17-1,69) e estresse (OR=1,41; IC 95%: 1,18-1,68). CONCLUSÃO: Vários fatores psicossociais foram associados à dor multirregional, indicando que estes devem ser abordados no manejo da dor.

Wound healing in weaning, adult, and old rats with provoked incisional hernias. A comparative study

Abstract Background: Incisional hernias are more frequent in adults than in children. It is hypothesized that a more efficient healing process in pediatric patients could explain this difference in incidence. Certain elements of healing such as neovascularization, degree of inflammation, percentage of mature and immature collagen, the proliferation of fibroblasts, and expression of certain genes could explain why healing in children is more efficient when compared to the adult and elderly populations. Materials and methods: Seventy-one rats of 3 different age groups (weaning, adult, and old) underwent surgery with 3 different incisions (vertical, oblique, and horizontal). During the procedure, the skin and abdominal wall of the animal were sectioned and only the skin was sutured to mimic incisional hernia in the animals. Four weeks after surgery, the rats were euthanized, their skin was removed, and the extent of scar tissue formed in the muscle opening was measured. In addition, samples of the scar tissue were collected for histological, immunohistochemical, and molecular analyzes. Nine rats served as controls. Results: Shorter-length hernias were formed in weaning rats when compared to old ones when the surgical incision was horizontal (p = 0.03). There was a greater proliferation of fibroblasts in rats in the younger age groups, regardless of the type of incision. The Lox gene was more expressed in weaning rats with vertical and oblique incisions. Conclusions: These differences could explain the better healing and lower incidence of hernias in the pediatric population, although this aspect requires further studies.

Microsurgical arterial anastomosis in young and adult rats: an evolutive and comparative study

Purpose: To evaluate the caliber of an arterial micro-anastomosis in the young growing animal using a continuous suture technique. Additionally, late morphological changes and blood flows distal to the anastomosis were evaluated. Methods: Seventy-four Wistar rats were submitted to laparotomy to access the aorta for blood flow measurement. The aorta was sectioned using microsurgery technique and an end-to-end anastomosis with continuous suture. After a period of six months to one year, the anastomosis was checked. Results: Regarding the size of the aortas, comparing the pre- and postoperative values, there was an increase of 13.33% in adult animals and 25% in young animals, without any difference in the blood flows. Conclusions: The arteries of young rats show signs of growth at the site of the anastomosis performed with continuous suture.

Os estudos das representações sociais sobre a velhice em contextos educacionais / The qualitive studies of social representations about old age in educational contexts

Este artigo apresenta um mapeamento de teses e dissertações que tiveram como objeto central de suas análises o processo de envelhecimento ou velhice, em contextos educacionais ou relacionados com a área da Educação. Os dados foram obtidos do Catálogo de Teses e Dissertações da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). O texto segue organizado em duas seções. Uma primeira, na qual são apresentadas as produções que ilustram os contextos iniciais em que a velhice é qualitativamente investigada nas Ciências Sociais e na Psicologia Social (com especial ênfase nas Representações Sociais). Por fim, o levantamento das teses e dissertações conforme as unidades temáticas elencadas e as considerações sobre o percurso das análises. A partir do material inventariado, foi possível perceber maneiras distintas de empreender o olhar sobre a velhice, especialmente em relação ao emprego da teoria e instrumentos metodológicos.(AU)

This article organizes theses and dissertations that had as their central goal of analysis the aging process or old age, in educational contexts or related to the area of Education. The data were obtained from Catálogo de Teses e Dissertações da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). The text is organized in two sections. The first one presents the papers that illustrate the initial contexts in which old age is qualitatively inves- tigated in Social Sciences and Social Psychology (with special emphasis on Social Representations). Finally, the second section selects the theses and dissertations according to the thematic units listed and the considerations on the course of the analysis. From the inventoried material, it was possible to perceive different ways of undertaking a look on old age, especially in relation to the use of theory and methodological instruments.(AU)

Cross-cultural adaptation and psychometric properties of the short version of COPSOQ II-Brazil

ABSTRACT OBJECTIVES Translate and culturally adapt the short version of Copenhagen Psychosocial Questionnaire II (COPSOQ II) into Brazilian Portuguese (COPSOQ II-Br) and evaluate its psychometric properties. METHODS Translation and cultural adaptation followed the standardized guidelines. Structural validity was assessed using exploratory factorial analysis. Test-retest reliability was evaluated by intraclass correlation coefficient (ICC3,1) and internal consistency by Cronbach's alpha. Floor and ceiling effect was considered acceptable if less than 15% of participants reported the lowest or highest scores. Measurement error was assessed by standard error of measurement (SEM), while construct validity was tested by correlating the COPSOQ II-Br, the Job Content Questionnaire and the Nordic Musculoskeletal Questionnaire. RESULTS The study evaluated a total of 211 civil servants and service providers in the test and 157 in the retest. After cross-cultural adaptation, the COPSOQ II-Br structure comprised seven domains and 11 dimensions. Most dimensions showed acceptable floor and ceiling effects, excepting "Work family conflicts" (floor effect of 26.1%), and "Meaning and commitment" and "Job satisfaction," with ceiling floor of 27.5% and 22.3%, respectively. Cronbach's alpha values reached the recommended levels (varied between 0.70 and 0.87). Test-retest reliability indicated that all dimensions had ICC between 0.71 and 0.81. SEM ranged from 0.6 to 2.2 and the construct validity showed good results with the tested instruments (significant positive and negative correlations). CONCLUSIONS All psychometric properties of the short version COPSOQ II-Br are suitable for use in Brazil. The instrument is thus validated and can be used by occupational health and human resources professionals to evaluate psychosocial working conditions.

Cardiac and renal effects of liver cirrhosis in a growing animal model

ABSTRACT Purpose: To assess the biochemical, histological, histomorphometric and molecular effects of biliary duct ligation (BDL) induced liver cirrhosis in the heart and kidneys. Methods: Thirty-two weaning rats (21 days old, 50-70 g) underwent BDL and were divided in four groups (euthanasia after two, four, six, and eight weeks, respectively) and compared to control groups. Results: The animals' hearts of group 3 were bigger than those of the control group (p=0.042), including thinner right ventricle wall, decreased internal diameter of ventricles, and increased perivascular collagen deposition in left ventricle, as well as increased interstitial collagen in right ventricle after six weeks. In the kidneys of groups 3 and 4, bilirubin impregnation in the tubules, hydropic degeneration, loss of nuclei and lack of plasmatic membrane limits were noted. Nitric oxide synthase (NOS) gene expressions were higher in group 1 (p=0.008), and endothelial nitric oxide synthase (eNOS) gene expressions were elevated in all experimental groups (p=0.008, p=0.001, p=0.022, and p=0.013, respectively). In the heart, a decreased expression of eNOS in group 1 (p=0.04) was observed. Conclusions: Liver cirrhosis leads to histological and histomorphometric alterations in the heart and kidneys, with changes in the NOS and eNOS gene expressions, that may suggest a role in the associated myocardial and renal manifestations.

Sepsis and cirrhosis in growing animals: description of a new experimental model and its pathological and immunological reliability

OBJECTIVES: In cirrhotic children, infection events and sepsis are more frequent and more severe due to immune dysfunction. The objectives of the current study were therefore to develop an experimental model of infection and sepsis in cirrhotic weaning growing rats, by the use of bile duct ligation (BDL) and cecal ligation and puncture (CLP). Additionally, the correlation of the clinico-histopathological data and serial cytokine levels in septic cirrhotic and non-cirrhotic animals was studied. METHODS: Young Wistar rats of age 21 days and of weight between 70-90 g were divided into 12 groups according to the surgical procedure performed: sham (sacrificed after 2 or 4 weeks), BDL (sacrificed after 2 or 4 weeks), CLP (2- or 4-week old animals sacrificed after 12 or 24 hours), BDL+CLP (2- or 4-week old animals sacrificed after 12 hours). Histopathological studies and determination of serum levels of cytokines IL-1 beta, IL-10, and TNF-alpha, for studies of systemic infection, were performed. Murine sepsis scores (MSS) based on the clinical aspects just before euthanasia were also included. RESULTS: A transitory increase in IL-1, IL-10, and TNF-alpha levels was observed, with different patterns according to the groups. Two-hit groups tended to present with higher values of serum cytokines and histopathological scores than their septic non-cirrhotic counterparts. There was a correlation between mortality rate and MSS (p<0.0001). CONCLUSION: The model is feasible and may be utilized in studies on liver cirrhosis and infection in growing animals.

Are there differences in the growth adaptation processes of growing and mature organism models of short bowel syndrome?

OBJECTIVES: The purpose of this study was to present an experimental model of short bowel syndrome (SBS) in weaning rats and to compare the adaptative mechanisms of the remaining bowel in weaning rats and adult animals by means of morphometric, histologic and molecular methods. METHODS: Twenty-four weaning rats were divided into 3 groups of 8 animals, one control group and two short bowel groups (euthanasia after 4 and 21 days), and were compared with similar adult groups. Morphometric evaluations of the animals and histopathological and molecular studies of the remaining bowel were performed. RESULTS: The weight of young rats increased after enterectomy, whereas that of adult rats decreased after enterectomy (p<0.0001). The ratio of intestinal length/body weight was significantly higher in weaning rats than in adults (p<0.002), showing that intestinal growth was more intense in weaning rats. Intestinal resection promoted increased thickness of the small bowel lamina propria (p=0.001) and reduced thickness of the colon lamina propria (p=0.04) in weaning rats relative to those in adults. In addition, intestinal resection promoted increased expression of the Bcl-xl gene (antiapoptotic) in adult animals compared with that in weaning rats (p=0.001). CONCLUSION: Morphometric, histological and molecular differences were shown in the adaptation processes of growing and mature organisms.

Immediate expression of c-fos and c-jun mRNA in a model of intestinal autotransplantation and ischemia-reperfusion in situ

OBJECTIVE: Intestinal ischemia-reperfusion injury occurs in several clinical conditions and after intestinal transplantation. The aim of the present study was to investigate the phenomena of apoptosis and cell proliferation in a previously described intestinal ischemia-reperfusion injury autograft model using immunohistochemical markers. The molecular mechanisms involved in ischemia-reperfusion injury repair were also investigated by measuring the expression of the early activation genes c-fos and c-jun, which induce apoptosis and cell proliferation. MATERIALS AND METHODS: Thirty adult male Wistar rats were subjected to surgery for a previously described ischemia-reperfusion model that preserved the small intestine, the cecum and the ascending colon. Following reperfusion, the cecum was harvested at different time points as a representative segment of the intestine. The rats were allocated to the following four subgroups according to the reperfusion time: subgroup 1: 5 min; subgroup 2: 15 min; subgroup 3: 30 min; and subgroup 4: 60 min. A control group of cecum samples was also collected. The expression of c-fos, c-jun and immunohistochemical markers of cell proliferation and apoptosis (Ki67 and TUNEL, respectively) was studied. RESULTS: The expression of both c-fos and c-jun in the cecum was increased beginning at 5 min after ischemia-reperfusion compared with the control. The expression of c-fos began to increase at 5 min, peaked at 30 min, and exhibited a declining tendency at 60 min after reperfusion. A progressive increase in c-jun expression was observed. Immunohistochemical analyses confirmed these observations. CONCLUSION: The early activation of the c-fos and c-jun genes occurred after intestinal ischemia-reperfusion injury, and these genes can act together to trigger cell proliferation and apoptosis. .

Effects of ischemic preconditioning in a pig model of large-for-size liver transplantation

OBJECTIVE: In most cases of pediatric liver transplantation, the clinical scenario of large-for-size transplants can lead to hepatic dysfunction and a decreased blood supply to the liver graft. The objective of the present experimental investigation was to evaluate the effects of ischemic preconditioning on this clinical entity. METHODS: Eighteen pigs were divided into three groups and underwent liver transplantation: a control group, in which the weights of the donors were similar to those of the recipients, a large-for-size group, and a large-for-size + ischemic preconditioning group. Blood samples were collected from the recipients to evaluate the pH and the sodium, potassium, aspartate aminotransferase and alanine aminotransferase levels. In addition, hepatic tissue was sampled from the recipients for histological evaluation, immunohistochemical analyses to detect hepatocyte apoptosis and proliferation and molecular analyses to evaluate the gene expression of Bax (pro-apoptotic), Bcl-XL (anti-apoptotic), c-Fos and c-Jun (immediate-early genes), ischemia-reperfusion-related inflammatory cytokines (IL-1, TNF-alpha and IL-6, which is also a stimulator of hepatocyte regeneration), intracellular adhesion molecule, endothelial nitric oxide synthase (a mediator of the protective effect of ischemic preconditioning) and TGF-beta (a pro-fibrogenic cytokine). RESULTS: All animals developed acidosis. At 1 hour and 3 hours after reperfusion, the animals in the large-for-size and large-for-size + ischemic preconditioning groups had decreased serum levels of Na and increased serum levels of K and aspartate aminotransferase compared with the control group. The molecular analysis revealed higher expression of the Bax, TNF-alpha, I-CAM and TGF-beta genes in the large-for-size group compared with the control and large-for-size + ischemic preconditioning groups. Ischemic preconditioning was responsible for an increase in c-Fos, IL-1, IL-6 and e-NOS ...

A simplified experimental model of large-for-size liver transplantation in pigs

OBJECTIVE: The ideal ratio between liver graft mass and recipient body weight for liver transplantation in small infants is unknown; however, if this ratio is over 4%, a condition called large-for-size may occur. Experimental models of large-for-size liver transplants have not been described in the literature. In addition, orthotopic liver transplantation is marked by high morbidity and mortality rates in animals due to the clamping of the venous splanchnic system. Therefore, the objective of this study was to create a porcine model of large-for-size liver transplantation with clamping of the supraceliac aorta during the anhepatic phase as an alternative to venovenous bypass. METHOD: Fourteen pigs underwent liver transplantation with whole-liver grafts without venovenous bypass and were divided into two experimental groups: the control group, in which the weights of the donors were similar to the weights of the recipients; and the large-for-size group, in which the weights of the donors were nearly 2 times the weights of the recipients. Hemodynamic data, the results of serum biochemical analyses and histological examination of the transplanted livers were collected. RESULTS: The mortality rate in both groups was 16.5% (1/7). The animals in the large-for-size group had increased serum levels of potassium, sodium, aspartate aminotransferase and alanine aminotransferase after graft reperfusion. The histological analyses revealed that there were no significant differences between the groups. CONCLUSION: This transplant method is a feasible experimental model of large-for-size liver transplantation. .