Use of ultrasound and targeted physiotherapy to manage nerve sutures placed under joint flexion: a case series.

BACKGROUND: Joint flexion to diminish the gap and avoid nerve grafts fell into disuse for decades, but recently attention for using this technique was regained. We report a case series of nerve suture under joint flexion, ultrasound monitoring, and physiotherapy. Our main objective was to determine how effective this multimodality treatment is. METHODS: A retrospective review of 8 patients treated with direct repair with joint flexion was done. Depending on the affected nerve, either the knee or the elbow was flexed intraoperatively to determine if direct suturing was possible. After surgery, the limb was held immobilized. Through serial ultrasounds and a physiotherapy program, the limb was fully extended. If a nerve repair rupture was observed, the patient was re-operated and grafts were used. RESULTS: Of the eight nerve sutures analyzed, four sustained a nerve rupture revealed by US at an early stage, while four did not show any sign of dehiscence. In the patients in whom the nerve suture was preserved, an early and very good response was observed. Ultrasound was 100% accurate at identifying nerve suture preservation. Early detection of nerve failure permitted early re-do surgery using grafts without flexion, ultimately determining good final results. CONCLUSIONS: We observed a high rate of dehiscence in our group of patients treated with direct repair and joint flexion. We believe this was due to an incorrect use of the immobilization device, excessive movement, or a broken device. In opposition to this, we observed that applying direct nerve sutures and joint flexion offers unusually good and fast results. If this technique is employed, it is mandatory to closely monitor suture status with US, together with physiotherapy providing progressive, US-guided extension of the flexed joint. If nerve rupture occurs, the close monitoring dictated by this protocol should ensure the timely application of a successful graft repair.

[Effect of a physical training program on the occurrence of exercise-induced arrhythmias in patients with heart disease]. / Efecto de un programa de entrenamiento físico en la ocurrencia de arritmias inducidas por el ejercicio en pacientes con cardiopatía.

Objective: Although physical training has been associated with an increase in survival, its role in reducing Exercise-induced arrhythmias (EIA) in patients with heart disease has not yet been dilucidated. We aim to compare the effect of physical training on the occurrence of EIA in patients with heart disease. Methodology: We evaluated a retroprospective and self-controlled cohort of patients older than 18 years with heart disease who entered the cardiac rehabilitation program of the National Institute of Cardiology Ignacio Chávez in México, during January 2015 to December 2016. In all patients, cardiovascular risk was stratified, including a cardiopulmonary exercise test, which was also performed at the end of the program. The occurrence of arrhythmias was evaluated before and after the training program. Results: 160 patients were analyzed, of which 126 (79%) were male. The proportion of patients who developed EIA in the first exercise testing was 56% and in the second one was 48%. In the first group 35% of patients did not developed EIA in the second test. Of the 71 patients who did not present EIA at the beginning of the program, twenty (28%) presented arrhythmias at second test. In relation to the effect of the training program on the occurrence of arrhythmias, we could observe that it had a protective effect, with a RR = 0.49 (95% CI: 0.35-0.67, p <0.001). Conclusion: In this study, physical training was associated with a decreased risk of developing EIA.

Effects of Farm Management Practices and Transport Time on Post-Mortem Changes of Longissimus lumborum Muscle Proteins in Suckling Goat Kids.

The combined effect of farm management practices, transport time, and ageing time on the electrophoretic changes of sarcoplasmic (SPP) and myofibrillar (MFP) protein fractions of goat kids was studied. A total of 64 suckling goat kids were withdrawn from two farms with "high" (GW) and "low" (DW) welfare-friendly management practices, and they were transported for 2 or 6 h immediately before slaughtering. Longissimus lumborum samples were obtained at 3, 8, and 21 days post-mortem, and muscle proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis SDS-PAGE. Both protein extracts displayed significant changes attributable to meat maturation. Managing conditions of kids in DW farms increased the post-mortem susceptibility of muscle proteins. Some MFP of Longissimus lumborum muscle, such as troponin T, as well as 26-30 and 35-37 kDa fractions were influenced significantly by deficient on-farm management, and therefore, these protein fragments might be considered as indicators of low-welfare on-farm management in goat kids.

Niños y adolescentes con enfermedades raras atendidos en consulta provincial de nutrición / Children and Adolescents with Rare Diseases Treated in Provincial Nutrition Consultation

RESUMEN Fundamento: el padecimiento de una enfermedad rara produce dificultades para su diagnóstico oportuno. Su pronóstico ensombrece cuando se asocia a desnutrición. Objetivo: caracterizar a niños y adolescentes con enfermedades raras y desnutrición, atendidos en consulta provincial de nutrición en Cienfuegos. Métodos: se realizó un estudio descriptivo, retrospectivo y transversal cuyo universo lo constituyeron los 13 niños y adolescentes con enfermedades raras y desnutrición de gravedad variable que acudieron ese periodo a consulta en el Hospital Pediátrico de Cienfuegos en el entre enero y junio del 2017. Los datos fueron obtenidos de la revisión de la historia clínica, la entrevista médica, valoración clínica y nutricional. Las variables analizadas fueron: edad, sexo, tipo de desnutrición, peso al nacer, hábitos alimentarios y alteraciones en aparatos y sistemas. Resultados: hubo predominio del sexo masculino con 9 (69,2 %) y del grupo mayor de 15 años, con 5 (38,4 %). La desnutrición moderada fue la que más se manifestó con 6 (46,1 %). Es representativo que 8 pacientes exhibieron un peso al nacer de más de 3 000 gramos, para un 61,5 %, mientras que los hábitos alimentarios adecuados se presentaron con más frecuencia, resaltan los pacientes evaluados como desnutridos moderados dentro del grupo. El 100 % de los pacientes presentan alteración musculo-esquelética. La baja talla fue la más frecuente de las afectaciones del crecimiento y desarrollo, con un 46,1 %. Los pacientes con las formas severas de desnutrición presentan una mayor expresividad de la sintomatología. Conclusión: predominó como el más afectado, el grupo de más de 15 años, el sexo masculino, el peso normal al nacer y los hábitos alimentarios adecuados. Fueron las alteraciones musculo-esqueléticas representativas en todos los casos.

ABSTRACT Foundation: suffering an uncommon disease causes difficulties for its timely diagnosis. Their prognosis overshadows when associated with malnutrition. Objective: to characterize children and adolescents with uncommon diseases and malnutrition, assisted in provincial nutritional consultation in Cienfuegos. Methods: a descriptive, retrospective and cross-sectional study whose universe consisted of 13 children and adolescents with uncommon diseases and malnutrition of variable severity who attended the Pediatric Hospital consultation of Cienfuegos. Data were obtained from clinical history review, medical interview and clinical and nutritional assessment. The following variables were analyzed: age, sex, type of malnutrition, birth weight, eating habits and alterations in apparatus and systems. The results were taken to a database for statistical processing by the SPSS program version 15,0. The results were presented in frequency and ratio tables of variables expressed in frequency number and in percentages. Results: there was predominance of the male sex with 9 (69,2 %) and of the group over 15 years old, with 5 (38,4 %). Moderate malnutrition was the frequent with 6 (46,1 %). It is representative that 8 patients exhibited a birth weight of more than 3,000 grams, for 61,5 %, while adequate eating habits occurred more frequently, highlighting the patients evaluated as moderate malnourished within the group. All of the patients have musculoskeletal disorders. The short stature was the most frequent of growth and development affectations, with 46,1 %. Patients with severe forms of malnutrition present a greater symptomatology expressivity. Conclusion: the most affected group was male sex over 15 years old, the normal weight at birth and the adequate eating habits prevailed. There were the representative musculoskeletal alterations in all cases.

Diagnosis of Superficial Mycoses by a Rapid and Effective PCR Method from Samples of Scales, Nails and Hair.

Superficial mycoses are the most frequently diagnosed affections of the stratum corneum of the skin, nails and hair. It is generally caused by the presence of yeasts and dermatophytes. Onychomycosis is the most common infection with an incidence of 80-90% in Europe generally produced by Trichophyton rubrum. The aim of this study is to compare the traditional diagnostic techniques of superficial mycoses with a homemade and wide-spectrum fungal polymerase chain reaction (PCR) technique that amplifies a specific region of the 18S ribosomal RNA (rRNA) directly from samples of scales, nails and hair. A total of 626 clinical samples (obtained in the Basurto University Hospital, Bilbao, Spain) were analysed by traditional culture, microscopy and PCR. DNA extraction was carried out by using an extraction buffer and bovine serum, and amplification of samples and performance of the PCR were checked by conventional agarose gel electrophoresis with subsequent sequencing of amplified samples. A total of 211 samples (34%) resulted in positive diagnosis with at least one of the two applied methods: culture (21%) and PCR (22%). Despite the low percentage of identification achieved by the sequencing technique (40%), the value contributed by the amplification of the 18S region of the rRNA was considered important in the identification as it showed a high predictive values for both positive and negative diagnoses (90.9% and 94.6%, respectively). The proposed PCR method has been confirmed as a complementary, rapid, and effective method in the diagnosis of superficial mycoses. Additionally, it reduces the time to obtain satisfactory results from 4 weeks to 7 h.

Calprotectin protects against experimental colonic inflammation in mice.

BACKGROUND AND PURPOSE: Calprotectin is a heterodimer composed of two myeloid-related proteins, S100A8 and S100A9, that is abundant in neutrophils and monocytes/macrophages. Faecal levels of calprotectin are used routinely to monitor inflammatory bowel disease activity. EXPERIMENTAL APPROACH: We aimed to assess the role of calprotectin in intestinal inflammation, using the dextran sulfate sodium model of colitis in mice. Calprotectin was administered (50 or 100 µg·day-1 ) by the intrarectal or by i.p. injection (50 µg·day-1 only). The condition of the mice was characterized by morphological and biochemical methods. KEY RESULTS: Intrarectal calprotectin protected significantly against colitis, as shown by lower levels of macroscopic and microscopic damage, colonic myeloperoxidase activity and decreased expression of TNFα and toll-like receptor 4. IL-17 production by spleen and mesenteric lymph node cells was reduced. Calprotectin had no effect on body weight loss or colonic thickening. There were no effects of calprotectin after i.p. injection. Calprotectin had virtually no effects in control, non-colitic mice. Calprotectin had almost no effect on the colonic microbiota but enhanced barrier function. Treatment of rat IEC18 intestinal epithelial cells in vitro with calprotectin induced output of the chemokines CXL1 and CCL2, involving the receptor for advanced glycation end products- and NFκB. CONCLUSION AND IMPLICATIONS: Calprotectin exerted protective effects in experimental colitis when given by the intrarectal route, by actions that appear to involve effects on the epithelium.

Association between premature ventricular complexes during exercise, long-term occurrence of life-threatening arrhythmia and mortality.

OBJECTIVE: Exercise-induced premature ventricular complexes (EiPVCs) are often considered as benign arrhythmias, although they are associated with a high risk of all-cause death in the general healthy population. However, an intermediate pathophysiological process remains unclear, particularly in patients with known cardiovascular disease. The aim of this study was to find an association between EiPVCs, the occurrence of life-threatening ventricular arrhythmias (LACO), and all-cause mortality in patients with cardiovascular disease. METHODS: This was an observational study of a cohort of patients with coronary artery disease (CAD) or idiopathic cardiomyopathy (ICM). Stress testing was performed as a part of the routine cardiovascular evaluation. The occurrence of EiPVCs was evaluated during exercise testing (ET). At follow-up, long-term occurrence of LACO was evaluated. A bivariate and multivariate analysis was performed. RESULTS: Out of the total of 1442 patients analysed, 700 (49%) had EiPVCs. During 14 years of following-up after ET, 106 LACO outcomes were observed. Long-term all-cause mortality was 4% (n=61). A bivariate analysis showed that patients with EiPVCs had an increased risk for LACO (RR=2.81, 95% CI; 1.9-4.3, P<.001), and for mortality (RR=2.1, CI95% 1.2-3.4, P<.01). Occurrence of LACO was also associated with a higher mortality risk (RR=5.7, 95% CI; 3.4-9.4, P<.001). After a post hoc analysis, LACO remained as a highly predictive variable for mortality. CONCLUSION: Patients with EiPVCs have a high risk of LACO and all-cause mortality. The presence of LACO could be an intermediate stage between EiPVCs and mortality in subjects with cardiovascular disease.

Association between premature ventricular complexes during exercise, long-term occurrence of life-threatening arrhythmia and mortality / Asociación entre ectopia ventricular en ejercicio, la ocurrencia de arritmias potencialmente mortales y mortalidad a largo plazo

Abstract Objective: Exercise-induced premature ventricular complexes (EiPVCs) are often considered as benign arrhythmias, although they are associated with a high risk of all-cause death in the general healthy population. However, an intermediate pathophysiological process remains unclear, particularly in patients with known cardiovascular disease. The aim of this study was to find an association between EiPVCs, the occurrence of life-threatening ventricular arrhythmias (LACO), and all-cause mortality in patients with cardiovascular disease. Methods: This was an observational study of a cohort of patients with coronary artery disease (CAD) or idiopathic cardiomyopathy (ICM). Stress testing was performed as a part of the routine cardiovascular evaluation. The occurrence of EiPVCs was evaluated during exercise testing (ET). At follow-up, long-term occurrence of LACO was evaluated. A bivariate and multivariate analysis was performed. Results: Out of the total of 1442 patients analysed, 700 (49%) had EiPVCs. During 14 years of following-up after ET, 106 LACO outcomes were observed. Long-term all-cause mortality was 4% (n = 61). A bivariate analysis showed that patients with EiPVCs had an increased risk for LACO (RR = 2.81, 95% CI; 1.9---4.3, P < .001), and for mortality (RR = 2.1, CI95% 1.2---3.4, P < .01). Occurrence of LACO was also associated with a higher mortality risk (RR = 5.7, 95% CI; 3.4---9.4, P < .001). After a post hoc analysis, LACO remained as a highly predictive variable for mortality. Conclusion: Patients with EiPVCs have a high risk of LACO and all-cause mortality. The presence of LACO could be an intermediate stage between EiPVCs and mortality in subjects with cardiovascular disease.

Resumen Objetivo: La extrasistolia ventricular inducida por ejercicio (EVIE) suele considerarse una arritmia benigna, sin embargo, ha sido asociada a mayor mortalidad en población general. Hasta hoy no se ha encontrado el proceso fisiopatológico involucrado, particularmente en pacientes con enfermedad cardiovascular. El objetivo del estudio fue establecer una asociación entre EVIE con la ocurrencia de arritmias ventriculares potencialmente malignas (APM) y letalidad a largo plazo, en sujetos con enfermedad cardiovascular. Método: Estudiamos una cohorte de pacientes con enfermedad coronaria o miocardiopatía dilatada, que realizaron una prueba de ejercicio al inicio del estudio. Inicialmente evaluamos la ocurrencia de EVIE, APM y letalidad a largo plazo y posteriormente se realizó un estudio bivariable y multivariable. Resultados: Se incluyeron 1,442 pacientes de los cuales 700 presentaron EVIE (49%). Durante 14 an˜os de seguimiento, 106 sujetos presentaron APM y la mortalidad total fue de 61 casos (4%). El estudio bivariable mostró que aquellos pacientes con EVIE tuvieron un riesgo de presentar APM de 2.81 (IC95% de 1.9 a 4.3, p < 0.001) y de mortalidad de 2.1 (IC95% de 1.2 a 3.4, p < 0.01). Los individuos con APM tuvieron mayor riesgo de mortalidad (RR= 5.7, IC95% de 3.4 a 9.4, p < 0.001). Después de un análisis multivariable, la APM continuó siendo una variable altamente predictiva de mortalidad. Conclusión: Los pacientes con EVIE tienen un riesgo elevado de presentar APM y de mortalidad a largo plazo. Los resultados sugieren que las APM podrían representar un estado intermedio entre la presencia de EVIE y la mortalidad en pacientes con enfermedad cardiovascular.

National Registry of Cardiac Rehabilitation Programs in Mexico II (RENAPREC II) / Registro Nacional de Programas de Rehabilitación Cardiaca en México II (RENAPREC II)

Abstract: Objective: The aim of this paper is to compare the state of Cardiac Rehabilitation Programs (CRP) in 2009 with 2015. Focus is directed on health care, training of health-providers, research, and the barriers to their implementation. Methods: All authors of RENAPREC-2009, and other cardiac rehabilitation leaders in Mexico were requested to participate. These centres were distributed into two groups: RENAPREC-2009 centres that participated in 2015, and the new CRP units. Results: In 2009 there were 14 centres, two of which disappeared and another two did not respond. CRP-units increased by 71% (n = 24), and their geographic distribution shows a centripetal pattern. The coverage of CRP-units was 0.02 centres per 100,000 inhabitants. Only 4.4% of eligible patients were referred to CRP, with a rate of 10.4 patients/100,000 inhabitants in 2015. The ratio of Clinical Cardiologists to Cardiac Rehabilitation Specialists was 94:1, and the ratio of Intervention Specialists to cardiac rehabilitation experts was 16:1. Cardiac rehabilitation activities and costs varied widely. Patient dropout rate in phase II was 12%. Several barriers were identified: financial crisis (83%), lack of skilled personnel (67%), deficient equipment (46%), inadequate areas (42%), and a reduced number of operating centres (38%). Conclusions: CRPs in Mexico are still in the process of maturing. Mexican CRP-centres have several strengths, like the quality of the education of the professionals and the multidisciplinary programs. However, the lack of referral of patients and the heterogeneity of procedures are still their main weaknesses.

Resumen: Objetivo: El propósito de este trabajo es comparar el estado actual de los programas de rehabilitación cardiaca (PRC) en México con el RENAPREC-2009, dirigido a la asistencia, docencia, investigación y barreras. Métodos: Se convocó a participar a todos los autores de RENAPREC-2009 y a otros líderes en rehabilitación cardiaca de México. Los centros fueron distribuidos en 2 grupos: los que participaron en el 2015 y las nuevas unidades de PRC. Resultados: En 2009 había 14 centros operativos, de los cuales 2 cerraron y 2 no respondieron. En 2015 se registraron 24 centros en total, representando un aumento neto del 71%. La distribución geográfica fue centrípeta. La cobertura fue de 0.02 centros/100,000 habitantes y de solamente un 4.4% de los pacientes elegibles (10.4 pacientes/100,000 habitantes). La relación cardiólogo clínico-rehabilitador cardiaco es de 94:1 y la de intervencionista-rehabilitador cardiaco es de 16:1. Las actividades realizadas y los costos de los PRC varían de forma importante de centro a centro. En promedio, el 12% de los pacientes en fase ii abandonaron el programa. Las principales barreras para el desarrollo de PRC fueron: económicas (83%), falta de personal capacitado (67%), falta de equipo (46%), áreas inadecuadas (42%) y un insuficiente número de centros operativos (38%). Conclusiones: Los PRC en nuestro país continúan en crecimiento. Se observan fortalezas como el nivel de docencia y el enfoque multidisciplinario, así como deficiencias en la homogeneidad de las actividades y la falta de referencia de la población elegible.

A synbiotic composed of Lactobacillus fermentum CECT5716 and FOS prevents the development of fatty acid liver and glycemic alterations in rats fed a high fructose diet associated with changes in the microbiota.

We investigated the effect of a high fructose diet (HFD) on Sprague Dawley rats and the impact of a synbiotic composed of Lactobacillus fermentum CECT5716 and fructooligosaccharides. Feeding the HFD for 5 weeks resulted in liver steatosis and insulin resistance but not obesity. These changes were associated with increased production of short-chain fatty acids and increased Bacteroidetes in feces, with an augmented Bacteroidetes/Firmicutes ratio, among other changes in the microbiota. In addition, barrier function was weakened, with increased LPS plasma levels. These data are consistent with increased fructose availability in the distal gut due to saturation of absorptive mechanisms, leading to dysbiosis, endotoxemia, hepatic steatosis, and insulin resistance. Treatment with the synbiotic prevented some of the pathological effects, so that treated rats did not develop steatosis or systemic inflammation, while dysbiosis and barrier function were greatly ameliorated. In addition, the synbiotic had hypolipidemic effects. The synbiotic composed by L. fermentum CECT5716 and fructooligosaccharides has beneficial effects in a model of metabolic syndrome induced by a HFD, suggesting it might be clinically useful in this type of condition, particularly considering that high fructose intake has been related to metabolic syndrome in humans.

National Registry of Cardiac Rehabilitation Programs in Mexico II (RENAPREC II).

OBJECTIVE: The aim of this paper is to compare the state of Cardiac Rehabilitation Programs (CRP) in 2009 with 2015. Focus is directed on health care, training of health-providers, research, and the barriers to their implementation. METHODS: All authors of RENAPREC-2009, and other cardiac rehabilitation leaders in Mexico were requested to participate. These centres were distributed into two groups: RENAPREC-2009 centres that participated in 2015, and the new CRP units. RESULTS: In 2009 there were 14 centres, two of which disappeared and another two did not respond. CRP-units increased by 71% (n=24), and their geographic distribution shows a centripetal pattern. The coverage of CRP-units was 0.02 centres per 100,000 inhabitants. Only 4.4% of eligible patients were referred to CRP, with a rate of 10.4 patients/100,000 inhabitants in 2015. The ratio of Clinical Cardiologists to Cardiac Rehabilitation Specialists was 94:1, and the ratio of Intervention Specialists to cardiac rehabilitation experts was 16:1. Cardiac rehabilitation activities and costs varied widely. Patient dropout rate in phase II was 12%. Several barriers were identified: financial crisis (83%), lack of skilled personnel (67%), deficient equipment (46%), inadequate areas (42%), and a reduced number of operating centres (38%). CONCLUSIONS: CRPs in Mexico are still in the process of maturing. Mexican CRP-centres have several strengths, like the quality of the education of the professionals and the multidisciplinary programs. However, the lack of referral of patients and the heterogeneity of procedures are still their main weaknesses.

[Comparison of peak oxygen uptake and VE/VCO2 slope, between children with and without heart failure]. / Comparación del consumo de oxígeno pico y la pendienteVE/VCO2 entre niños con y sin insuficiencia cardiaca (IC).

Heart failure is a health problem associated with disability and mortality. Physicians may stratify the risk of adult patients with heart failure using a cardiopulmonary exercise testing. Until now, in childhood this evaluation has been poorly used. The purpose of this study is to compare the peak oxygen uptake and minute ventilation/carbon dioxide production slope among children with heart failure versus children without heart disease (control). METHODS: Thirty-eight children with heart failure were compared with 194 children without heart disease. All of them performed cardiopulmonary exercise testing using a symptom-limited ramp protocol. Differences between groups were compared using Chi-squared test, Student's t test, or ANOVA. Any value of p < 0.05 was considered significant. RESULTS: Children with heart failure were older, taller, and with a higher prevalence of male gender. This group had also a lower peak oxygen uptake (27 ± 10 ml O2/kg/min) compared to the control group (37 ± 10 ml O2/kg/min); p < 0.001. The minute ventilation/carbon dioxide production was higher in the heart failure group (31 ± 4) than in controls (28 ± 6); p < 0.001. CONCLUSION: Children with heart failure showed lower peak oxygen uptake and higher minute ventilation/carbon dioxide production slope than the control group.

The glucocorticoid budesonide has protective and deleterious effects in experimental colitis in mice.

Glucocorticoids are widely used for the management of inflammatory bowel disease, albeit with known limitations for long-term use and relevant adverse effects. In turn, they have harmful effects in experimental colitis. We aimed to explore the mechanism and possible implications of this phenomenon. Regular and microbiota depleted C57BL/6 mice were exposed to dextran sulfate sodium (DSS) to induce colitis and treated with budesonide. Colonic inflammation and animal status were compared. In vitro epithelial models of wound healing were used to confirm the effects of glucocorticoids. Budesonide was also tested in lymphocyte transfer colitis. Budesonide (1-60µg/day) exerted substantial colonic antiinflammatory effects in DSS colitis. At the same time, it aggravated body weight loss, increased rectal bleeding, and induced general deterioration of animal status, bacterial translocation and endotoxemia. As a result, there was an associated increase in parameters of sepsis, such as plasma NOx, IL-1ß, IL-6, lung myeloperoxidase and iNOS, as well as significant hypothermia. Budesonide also enhanced DSS induced colonic damage in microbiota depleted mice. These effects were correlated with antiproliferative effects at the epithelial level, which are expected to impair wound healing. In contrast, budesonide had significant but greatly diminished deleterious effects in noncolitic mice or in mice with lymphocyte transfer colitis. We conclude that budesonide weakens mucosal barrier function by interfering with epithelial dynamics and dampening the immune response in the context of significant mucosal injury, causing sepsis. This may be a contributing factor, at least in part, limiting clinical usefulness of corticoids in inflammatory bowel disease.

Germ-free and Antibiotic-treated Mice are Highly Susceptible to Epithelial Injury in DSS Colitis.

BACKGROUND AND AIMS: Intestinal microbiota is required to maintain immune homeostasis and intestinal barrier function. At the same time, intraluminal bacteria are considered to be involved in inflammatory bowel disease and are required for colitis induction in animal models, with the possible exception of dextran sulphate sodium [DSS] colitis. This study was carried out to ascertain the mechanism underlying the induction of colitis by DSS in the absence of bacteria. METHODS: Conventional and germ-free [GF] Naval Medical Research Institute [NMRI] mice were used, plus conventional mice treated with an antibiotic cocktail to deplete the intestinal microbiota ['pseudo-GF' or PGF mice]. The differential response to DSS was assessed. RESULTS: Conventional mice developed DSS-induced colitis normally, whereas GF mice showed only minimal inflammation [no colonic thickening, lower myeloperoxidase activity, IL-6, IL-17, TNF-α, and IFN-γ secretion by splenocytes and mesenteric cell cultures, etc.]. However, these mice suffered enhanced haemorrhage, epithelial injury and mortality as a consequence of a weakened intestinal barrier, as shown by lower occludin, claudin 4, TFF3, MUC3, and IL-22. In contrast, PGF mice had a relatively normal, albeit attenuated, inflammatory response, but were less prone to haemorrhage and epithelial injury than GF mice. This was correlated with an increased expression of IL-10 and Foxp3 and preservation barrier-related markers. CONCLUSIONS: We conclude that enteric bacteria are essential for the development of normal DSS-induced colitis. The absence of microbiota reduces DSS colonic inflammation dramatically but it also impairs barrier function, whereas subtotal microbiota depletion has intermediate effects at both levels.

Fructooligosaccharides exert intestinal anti-inflammatory activity in the CD4+ CD62L+ T cell transfer model of colitis in C57BL/6J mice.

PURPOSE: Fructooligosaccharides (FOS) are used as functional foods due to their prebiotic effects. Intestinal anti-inflammatory activity has been established in most, but not all, studies in animal models of colitis, using mainly chemically induced inflammation. Our goal was to test the effect of FOS (degree of polymerization 2-8) in the chronic, lymphocyte-driven CD4+ CD62L+ T cell transfer model of colitis. METHODS: Colitis was induced by transfer of CD4+ CD62L+ T cells to C57BL/6J Rag1(-/-) mice. FOS (75 mg day(-1)) was administered by gavage as a post-treatment. Three groups were established: non-colitic (NC), colitic control (C, CD4+ CD62L+ transferred mice treated with vehicle) and colitic+FOS (C+FOS, similar but treated with FOS). Mice were killed after 13 days. RESULTS: Treatment of mice with FOS ameliorated colitis, as evidenced by an increase in body weight, a lesser myeloperoxidase and alkaline phosphatase activities, a lower secretion of proinflammatory cytokines by mesenteric lymph node cells ex vivo (IFN-γ, IL-17, and TNF-α), and a higher colonic expression of occludin (C+FOS vs. C, p < 0.05). Increased relative abundance of lactic acid bacteria was observed in FOS-treated mice (p < 0.05). CONCLUSIONS: FOS exert intestinal anti-inflammatory activity in T lymphocyte-dependent colitis, suggesting it may be useful in the management of inflammatory bowel disease in appropriate conditions.

Impact of the NOGG and NOF Guidelines on the Indication of Bone Mineral Density in Routine Clinical Practice.

The universal screening for osteoporosis by bone mineral density (BMD) is not feasible because of its unfavorable cost-benefit due to its low sensitivity. The aim of the present study was to estimate the population and economic impact of the diagnostic criteria of the National Osteoporosis Guideline Group (NOGG) and the National Osteoporosis Foundation (NOF) and assess the appropriateness of the BMD tests performed in routine clinical practice. A cross-sectional study was conducted in individuals referred for BMD testing who were not receiving antiresorptive therapy. The absolute risk of major and hip fracture was calculated using the British formula of the Fracture Risk Assessment Tool. NOGG and NOF guidelines diagnostic thresholds interventions were used. A total of 640 individuals were included, of which 95% were women, with a median age of 59.4 years (interquartile range = 14). When applying the NOGG criteria, BMD testing was recommended in 32.3% of the individuals, whereas this percentage increased to 75.6% with the NOF guidelines (p < 0.05). Regarding the appropriateness of the BMD tests performed, 31.9% were deemed appropriate according to both the NOGG and NOF guidelines, whereas 23.9% were considered inappropriate. In conclusion, the application of the NOGG and NOF guidelines led to a decrease in BMD indications, reducing costs and improving efficiency in the diagnostic management of osteoporosis, although variability exists between the guidelines.

Intestinal anti-inflammatory activity of apigenin K in two rat colitis models induced by trinitrobenzenesulfonic acid and dextran sulphate sodium.

Flavonoids are polyphenolic compounds that are widespread in nature, and consumed as part of the human diet in significant amounts. The aim of the present study was to test the intestinal anti-inflammatory activity of apigenin K, a soluble form of apigenin, in two models of rat colitis, namely the trinitrobenzenesulfonic acid (TNBS) model and the dextran sulphate sodium (DSS) model. Apigenin K (1, 3 and 10 mg/kg; by the oral route; n 4-6 per group) was administered as a pre-treatment to rats with TNBS and DSS colitis, and colonic status was checked by macroscopic and biochemical examination. Apigenin K pre-treatment resulted in the amelioration of morphological signs and biochemical markers in the TNBS model. The results demonstrated a reduction in the inflamed area, as well as lower values of score and colonic weight:length ratio compared with the TNBS group. Myeloperoxidase (MPO) activity was reduced by 30 % (P< 0·05). Moreover, apigenin K pre-treatment ameliorated morphological signs and biochemical markers in the DSS model. Thus, macroscopic damage was significantly reduced and the colonic weight:length ratio was lowered by approximately 10 %, while colonic MPO and alkaline phosphatase activities were decreased by 35 and 21 %, respectively (P< 0·05). Apigenin K pre-treatment also tended to normalise the expression of a number of colonic inflammatory markers (e.g. TNF-α, transforming growth factor-ß, IL-6, intercellular adhesion molecule 1 or chemokine (C-C motif) ligand 2). In conclusion, apigenin K is found to have anti-inflammatory effects in two preclinical models of inflammatory bowel disease.

Rutin has intestinal antiinflammatory effects in the CD4+ CD62L+ T cell transfer model of colitis.

Rutin, one of the most abundant flavonoids in nature, has been shown to exert intestinal antiinflammatory effects in experimental models of colitis. Our aim was to study the antiinflamatory effect of rutin in the CD4+ CD62L+ T cell transfer model of colitis, one of the closest to the human disease. Colitis was induced by transfer of CD4+ CD62L+ T cells to Rag1(-/-) mice. Rutin was administered by gavage as a postreatment. Treatment with rutin improved colitis at the dose of 57mg/kg/day, while no effect was noted with 28.5mg/kg/day. Therapeutic benefit was evidenced by a reduced disease activity index, weight loss and damage score, plus a 36% lower colonic myeloperoxidase and a 54% lower alkaline phosphatase activity. In addition, a decreased secretion of proinflammatory cytokines (IFNγ and TNFα) by mesenteric lymph node cells was observed ex vivo. The colonic expression of proinflammatory genes, including IFNγ, TNFα, CXCL1, S100A8 and IL-1ß, was significantly reduced by more than 80% with rutin as assessed by RT-qPCR. Flavonoid treated mice exhibited decreased activation of splenic CD4+ cells (STAT4 phosphorylation and IFNγ expression) and reduced plasma cytokine levels. This effect was also apparent in mucosal lymphocytes based on reduced STAT4 phosphorylation. The protective effect was comparable to that of 3mg/kg/day budesonide. Rutin had no effect on splenocytes or murine T cells in vitro, while its aglycone, quercetin, exhibited a concentration dependent inhibition of proinflammatory cytokines, including IFNγ. Rutin but not quercetin showed vectorial basolateral to apical transport in IEC18 cells, associated with reduced biotransformation. We conclude that rutin exerts intestinal antiinflammatory activity in chronic, T lymphocyte dependent colitis via quercetin release and actions involving mucosal and lymph node T cells. Our results suggest that rutin may be useful in the management of inflammatory bowel disease in appropriate dosage conditions.

Active hexose correlated compound exerts therapeutic effects in lymphocyte driven colitis.

Active hexose correlated compound (AHCC) is a commercial extract of Basidiomycetes fungi enriched in oligosaccharides that is used as a human nutritional supplement for various purposes in humans. Our aim was to study the anti-inflammatory effect of AHCC in the CD4+ CD62L(+) T cell transfer model of colitis, considered one of the closest to the human disease. Colitis was induced by transfer of CD4(+) CD62L(+) T cells to recombination activating gene 1(-/-) mice. AHCC (75 mg/d) was administered by gavage as a post-treatment. Three groups were established: noncolitic, colitic (CD4(+) CD62L(+) transferred mice treated with vehicle), and AHCC (colitic treated with AHCC). AHCC improved colitis, as evidenced by a 24% lower colonic myeloperoxidase and a 21% lower alkaline phosphatase activity. In addition, a decreased secretion of proinflammatory genes assessed by RT-qPCR was observed, particularly TNF-α and IL-1ß. Ex vivo mesenteric lymph node cells obtained from AHCC treated mice exhibited a fully normalized production of IL-6, IL-17, and IL-10 (p < 0.05). Also, AHCC treated mice exhibited decreased STAT4 and IκB-α phosphorylation in splenic CD4(+) cells. Our data provide validation of AHCC colonic anti-inflammatory activity in a chronic, T cell driven model of inflammatory bowel disease.

Prebiotic oligosaccharides directly modulate proinflammatory cytokine production in monocytes via activation of TLR4.

SCOPE: Prebiotic oligosaccharides are currently used in a variety of clinical settings for their effects on intestinal microbiota. Here, we have examined the direct, microbiota independent, effects of prebiotics on monocytes and T lymphocytes in vitro. METHODS AND RESULTS: Prebiotics generally evoked cytokine secretion (TNF-α, IL-6, and IL-10) by mouse splenocytes but inhibited LPS -induced IFN-γ and IL-17 release. Inulin was found to enhance LPS-induced IL-10 secretion. Splenocytes from TLR4(-/-) (where TLR is Toll-like receptor) mice showed a markedly depressed response. Conversely, in both basal and LPS-stimulated conditions, prebiotic inhibition of IFN-γ levels was preserved. These results suggested a predominant effect on monocytes via TLR4 ligation and possible inhibition of T cells. Hence, we studied the modulation of primary rat monocytes and T lymphocytes, focusing on fructooligosaccharides (FOS) and inulin. In monocytes, FOS and inulin induced TNF-α, growth-regulated oncogene α, and IL-10, but not IL-1ß release. The NF-κB inhibitor Bay 11-7082 fully prevented these effects. Pharmacological evidence also indicated a significant involvement of mitogen-activated protein kinase and phosphatidylinositol-3-kinase. There was little effect on T cells. FOS and inulin also generally increased TNF-α, IL-1ß, and IL-10, but not IL-8, in human peripheral blood monocytes. CONCLUSION: We conclude that prebiotics may act as TLR4 ligands or as indirect TLR4 modulators to upregulate cytokine secretion in monocytes.