Short-term impact of extreme temperatures, relative humidity and air pollution on emergency hospital admissions due to kidney disease and kidney-related conditions in the Greater Madrid area (Spain).

While some studies report a possible association between heat waves and kidney disease and kidney-related conditions, there still is no consistent scientific consensus on the matter or on the role played by other variables, such as air pollution and relative humidity. Ecological retrospective time series study 01-01-2013 to 31-12-2018). Dependent variables: daily emergency hospitalisations due to kidney disease (KD), acute kidney injury (AKI), lithiasis (L), dysnatraemia (DY) and hypovolaemia (HPV). Independent variables: maximum and minimum daily temperature (Tmax, Tmin, °C), and daily relative humidity (RH, %). Other variables were also calculated, such as the daily temperature for risk of kidney disease (Theat, °C) and low daily hazardous relative humidity (HRH%). As variables of air pollution, we used the daily mean concentrations of PM10, PM2.5, NO2 and O3 in µg/m3. Based on these, we then calculated their daily excesses over World Health Organisation (WHO) guideline levels (hPM10, hPM2.5, hNO2 and hO3 respectively). Poisson family generalised linear models (GLMs) (link = log) were used to calculate relative risks (RRs), and attributable risks and attributable admissions. In the models, we controlled for the covariates included: seasonalities, trend, autoregressive component, day of the week, month and year. A statistically significant association was found between Theat and all the dependent variables analysed. The greatest AKI disease burden was attributable to Theat (2.2 % (1.7, 2.6) of attributable hospital admissions), followed by hNO2 (1.7 % (0.9, 3.4)) and HRH (0.8 (0.6, 1.1)). In the case of hypovolaemia and dysnatraemia, the greatest disease burden again corresponded to Theat, with 6.9 % (6.2, 7.6) and 5.7 (4.8, 6.6) of attributable hospital admissions respectively. Episodes of extreme heat exacerbate daily emergency hospital admissions due to kidney disease and kidney-related conditions; and attributable risks are likewise seen for low relative humidity and high ozone levels.

[Influence of social inequalities in health on the mortality of the rural and urban population in Spain, 2007-2013]. / Influencia de las desigualdades sociales en salud en la mortalidad de la población rural y urbana en España, 2007-2013.

OBJECTIVE: Describe patterns of cause of death by gender, age and territory. The causes of mortality and their possible relationships with health inequalities in the rural and urban population in Spain. MATERIAL AND METHODS: An ecological study of the mortality data of the Spanish population between 2007 and 2013 was carried out. Mortality data were obtained from the National Statistics Institute (INE). To be able to present the data in the grouped and simplified form, the ICD-10 classification detected by the INE was coded with the classification of Disease Load defined by Murray and López (1996), reduced to 21 categories of diseases. The territory variable has been determined: rural / urban, taking as a reference the cut that establishes that populations over 10,000 inhabitants are urban environments and those that are below rural environments. The chi-square test and the Bonferroni-corrected z test for qualitative factors and the Student's t-test for quantitative factors were considered. RESULTS: The results showed than the main causes of death were cardiovascular diseases with 31% (844,010) and malignant tumors with 26.7% (724,889), neuropsychiatric with 8.8% (238,330) and respiratory with 8.7% (235,448). In the rural setting, for the male gender, 52.7% (366,053) died, while 51.3% (995,470) died in the urban environment. Regarding the female gender, the cases of deaths were 47.3% (329,063) in the rural environment and 48.7% (9545.188) in the urban. Related to age, the means of the ages associated with a later death were nutritional conditions with 85.62 years. As for the earliest ages, obviating those related to the deaths of newborns, were the congenital anomalies with 25.37 years of means. CONCLUSIONS: Mortality differences were found between the 3axes of social inequality in health-age, gender and territory. Therefore, we can say that social determinants condition our life expectancy.

Glucagon-like peptide-1 improves beta-cell antioxidant capacity via extracellular regulated kinases pathway and Nrf2 translocation.

Oxidative stress plays an important role in the development of beta-cell dysfunction and insulin resistance, two major pathophysiological abnormalities of type 2 diabetes. Expression levels of antioxidant enzymes in beta cells are very low, rendering them more susceptible to damage caused by reactive oxygen species (ROS). Although the antioxidant effects of glucagon-like peptide-1 (GLP-1) and its analogs have been previously reported, the exact mechanisms involved are still unclear. In this study, we demonstrated that GLP-1 was able to effectively inhibit oxidative stress and cell death of INS-1E beta cells induced by the pro-oxidant tert-butyl hydroperoxide (tert-BOOH). Incubation with GLP-1 enhanced cellular levels of glutathione and the activity of its related enzymes, glutathione-peroxidase (GPx) and -reductase (GR) in beta cells. However, inhibition of ERK, but not of the PI3K/AKT pathway abolished, at least in part, the antioxidant effect of GLP-1. Moreover, ERK activation seems to be protein kinase A (PKA)-dependent because inhibition of PKA with H-89 was sufficient to block the GLP-1-derived protective effect on beta cells. GLP-1 likewise increased the synthesis of GR and favored the translocation of the nuclear transcription factor erythroid 2p45-related factor (Nrf2), a transcription factor implicated in the expression of several antioxidant/detoxificant enzymes. Glucose-stimulated insulin secretion was also preserved in beta-cells challenged with tert-BOOH but pre-treated with GLP-1, probably through the down-regulation of the mitochondrial uncoupling-protein2 (UCP2). Thus, our results provide additional mechanisms of action of GLP-1 to prevent oxidative damage in beta cells through the modulation of signaling pathways involved in antioxidant enzyme regulation.

Evaluación por competencias de los estudiantes de Enfermería a través de Aprendizaje Basado en Problemas / Nursing student's competency assessment through Problem Based Learning

Introducción: El objetivo fue evaluar la adquisición de competencias del alumnado de enfermería aplicando Aprendizaje Basado en Problemas. Métodos: Estudio descriptivo y transversal. La muestra incluyó 23 grupos de estudiantes de enfermería de la Universidad de Valladolid que siguieron la metodología Aprendizaje Basado en Problemas, durante un cuatrimestre académico del año 2012. Los instrumentos utilizados fueron cuestionarios de evaluación a pares, autoevaluación y evaluación a la tutora, determinándose las proporciones de las respuestas; observación sistemática de las tutorías; análisis de contenido del informe escrito sobre un caso clínico realizado por cada grupo de alumnos y observación de la comunicación oral. Resultados: Se desarrollaron habilidades comunicativas, uso de herramientas tecnológicas, autonomía, trabajo colaborativo y razonamiento clínico. Conclusiones: El Aprendizaje Basado en Problemas permite evaluar competencias transversales en estudiantes de enfermería utilizando distintos instrumentos. La participación de los alumnos en la evaluación fortalece actitudes (saber ser) como la honestidad, la responsabilidad y la autonomía.

Introduction: The objective was to assess competency acquisition in nursing students using the Problem Based Learning approach. Methods: Transversal and descriptive study with a sample of 23 groups of nursing students from the Universidad de Valladolid who followed the Problem Based Learning methodology during an academic four month period in the year of 2012. The instruments used were pair evaluation questionnaires, self-evaluation, and preceptor evaluation to assess responses proportions, tutoring observation, clinical case written inform content analysis, and oral communication observation. Results: Improvements in communication skills development, technological tools use, autonomy, collaborative work, and clinical reasoning. Conclusions: The Problem Based Learning allows the assessment of transversal competencies in nursing students using different instruments. students participation in the assessment process strengthens attitudes (know being) such as honesty, responsibility, and autonomy.

Protein tyrosine phosphatase 1B modulates GSK3ß/Nrf2 and IGFIR signaling pathways in acetaminophen-induced hepatotoxicity.

Acute hepatic failure secondary to acetaminophen (APAP) poisoning is associated with high mortality. Protein tyrosine phosphatase 1B (PTP1B) is a negative regulator of tyrosine kinase growth factor signaling. In the liver, this pathway confers protection against injury. However, the involvement of PTP1B in the intracellular networks activated by APAP is unknown. We have assessed PTP1B expression in APAP-induced liver failure in humans and its role in the molecular mechanisms that regulate the balance between cell death and survival in human and mouse hepatocytes, as well as in a mouse model of APAP-induced hepatotoxicity. PTP1B expression was increased in human liver tissue removed during liver transplant from patients for APAP overdose. PTP1B was upregulated by APAP in primary human and mouse hepatocytes together with the activation of c-jun (NH2) terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38 MAPK), resulting in cell death. Conversely, Akt phosphorylation and the antiapoptotic Bcl2 family members BclxL and Mcl1 were decreased. PTP1B deficiency in mouse protects hepatocytes against APAP-induced cell death, preventing glutathione depletion, reactive oxygen species (ROS) generation and activation of JNK and p38 MAPK. APAP-treated PTP1B(-/-) hepatocytes showed enhanced antioxidant defense through the glycogen synthase kinase 3 (GSK3)ß/Src kinase family (SKF) axis, delaying tyrosine phosphorylation of the transcription factor nuclear factor-erythroid 2-related factor (Nrf2) and its nuclear exclusion, ubiquitination and degradation. Insulin-like growth factor-I receptor-mediated signaling decreased in APAP-treated wild-type hepatocytes, but was maintained in PTP1B(-/-) cells or in wild-type hepatocytes with reduced PTP1B levels by RNA interference. Likewise, both signaling cascades were modulated in mice, resulting in less severe APAP hepatotoxicity in PTP1B(-/-) mice. Our results demonstrated that PTP1B is a central player of the mechanisms triggered by APAP in hepatotoxicity, suggesting a novel therapeutic target against APAP-induced liver failure.

[Comparison of the post-surgical analgesic effectiveness of tibial (at internal malleolus level) and common peroneal nerve block with infiltration of the surgical wound in Outpatient Surgery of the hallux valgus]. / Comparación de la eficacia analgésica postoperatoria del bloqueo de los nervios tibial (a nivel de maléolo interno) y peroneo común con la infiltración de la herida quirúrgica en cirugía ambulatoria del hallux valgus.

INTRODUCTION: To compare the post-operative analgesic effectiveness of blocking the posterior tibial and the common peroneal nerves against that of wound infiltration using local anaesthesia, in ambulatory surgery of hallux valgus. MATERIAL AND METHODS: A randomised clinical study was conducted on ambulatory patients subjected to Hallux valgus surgery, assigned into two groups: BNP: peripheral nerve blockage: posterior tibial and the common peroneal with 80mg of lidocaine, 100mg of mepivacaine and 25mg of levobupivacaine. INF: surgical wound infiltration with 50mg of levobupivacaine. The following aspects were evaluated during the first 24h after surgery: pain level using a visual analogue scale (VAS), the need to use rescue analgesia, and the incidence of secondary effects and readmissions due to pain. RESULTS: A total of 111 Patients were included (55 BNP, 56 INF), 93 per cent were women and the average age was 59 (SD10) years. The average VAS score in the first 24h was 2.9 (SD1.7) for the BNP group and 2.7 (SD1.6) for the INF group (P=.62). Less than half (42%) of patients needed rescue anaesthetic with tramadol, with no significant differences between the groups (P=.28). A 33 per cent had secondary postoperative effects were observed in 33% of cases, with a significant difference between INF and BNP (P=.01). One patient from INF group, had to be admitted for pain. CONCLUSIONS: The peripheral nerve block and wound infiltration are valid techniques for controlling pain at home after ambulatory surgery of hallux valgus, therefore both methods appear to be safe in an outpatient setting.

[Clinical phenotypes associated with selective IgA deficiency: a review of 330 cases and a proposed follow-up protocol]. / Fenotipos clínicos asociados a la deficiencia selectiva de IgA: revisión de 330 casos y propuesta de un protocolo de seguimiento.

UNLABELLED: Selective IgA deficiency is the most common Primary Immune Deficiency. Only a small proportion of these patients present during childhood, but this proportion increases over the years, and may be associated with an IgG subclass deficiency with increased susceptibility to respiratory and digestive tract infections. During childhood, IgA deficient patients may also refer to symptoms related to allergic and autoimmune diseases or tumours. AIMS: To describe the relationship of selective IgA deficiency with infections, allergic diseases, autoimmune disorders and tumours. To investigate the presence of other immune disorders associated with selective IgA deficiency. To suggest a follow-up protocol for these patients. METHODS: Retrospective study of paediatric patients (<18 years) being followed-up in the Clinical Immunology Department between 1992 and 2007, as well as laboratory records with IgA values below 50mg/L. Clinical records were reviewed (frequency and intensity of diseases associated with selective IgA deficiency) along with immunology tests performed. RESULTS: A total of 330 paediatric patients were identified with a selective IgA deficiency: 39 (11.8%) suffered from recurrent ear infections (2 developed secondary deafness), 58 (17.5%) from recurrent upper respiratory tract infections, and 20 patients (6%) from recurrent pneumonia, 6 of whom developed secondary bronchiectasis and 2 underwent a lobectomy. A relationship with atopic disease was found in 62 (18.78%) of patients. Regarding digestive disorders, chronic diarrhoea was found in 21 (6.5%), coeliac disease in 22 (6.6%), and persistently high plasma transaminases in 3. Autoimmune manifestations were found in 38 (11.5%), juvenile chronic arthritis, type 1 diabetes, vitiligo, cytopenia, and Crohn's disease, amongst others). Tumours were identified in 5 (1.5%). An IgG sub-class deficiency was found in 5 patients (4%), and 6 patients had a confirmed deficiency in antibody production. CONCLUSIONS: In our cohort, 56.6% of patients with IgA deficiency showed other comorbidities which were, in decreasing frequency: recurrent infections (respiratory and ear infections), allergic diseases, autoimmunity and tumours. Some patients will develop a more severe humoral defect (IgG subclass deficiency with or without antibody deficiency).

Compromised gastrointestinal integrity in pigtail macaques is associated with increased microbial translocation, immune activation, and IL-17 production in the absence of SIV infection.

Pigtail macaques (PTMs) rapidly progress to AIDS after simian immunodeficiency virus (SIV) infection. Given the strong association between human immunodeficiency virus (HIV) and SIV disease progression and microbial translocation and immune activation, we assessed whether high basal levels of immune activation and microbial translocation exist in PTMs. We found that before SIV infection, PTMs had high levels of microbial translocation that correlated with significant damage to the structural barrier of the gastrointestinal tract. Moreover, this increased microbial translocation correlated with high levels of immune activation and was associated with high frequencies of interleukin-17-producing T cells. These data highlight the relationship among mucosal damage, microbial translocation and systemic immune activation in the absence of SIV replication, and underscore the importance of microbial translocation in the rapid course of disease progression in SIV-infected PTMs. Furthermore, these data suggest that PTM may be an ideal model to study therapeutic interventions aimed at decreasing microbial translocation-induced immune activation.

Mitochondrial bioenergetics and dynamics interplay in complex I-deficient fibroblasts.

BACKGROUND: Complex I (CI) deficiency is the most frequent cause of OXPHOS disorders. Recent studies have shown increases in reactive oxygen species (ROS) production and mitochondrial network disturbances in patients' fibroblasts harbouring mutations in CI subunits. OBJECTIVES: The present work evaluates the impact of mutations in the NDUFA1 and NDUFV1 genes of CI on mitochondrial bioenergetics and dynamics, in fibroblasts from patients suffering isolated CI deficiency. RESULTS: Decreased oxygen consumption rate and slow growth rate were found in patients with severe CI deficiency. Mitochondrial diameter was slightly increased in patients' cells cultured in galactose or treated with 2'-deoxyglucose without evidence of mitochondrial fragmentation. Expression levels of the main proteins involved in mitochondrial dynamics, OPA1, MFN2, and DRP1, were slightly augmented in all patients' cells lines. The study of mitochondrial dynamics showed delayed recovery of the mitochondrial network after treatment with the uncoupler carbonyl cyanide m-chlorophenyl hydrazone (cccp) in patients with severe CI deficiency. Intracellular ROS levels were not increased neither in glucose nor galactose medium in patients' fibroblasts. CONCLUSION: Our main finding was that severe CI deficiency in patients harbouring mutations in the NDUFA1 and NDUFV1 genes is linked to a delayed mitochondrial network recovery after cccp treatment. However, the CI deficiency is neither associated with massive mitochondrial fragmentation nor with increased ROS levels. The different genetic backgrounds of patients with OXPHOS disorders would explain, at least partially, differences in the pathophysiological manifestations of CI deficiency.

Living donor liver transplantation: usefulness of hemostatic and prothrombotic screening in potential donors.

Bleeding and thrombosis are serious complications of living donor liver transplantation (LDLT). The aim of this paper was to describe the results of a screening for coagulation disorders, including for thrombophilic factors, in potential living liver graft donors and to evaluate thrombotic and bleeding events in donors and recipients, during and after the procedure. From January 2001 to January 2007, 41 LDLTs were performed at our institution. We performed systematic screening for bleeding or prothrombotic states among 188 potential donors, 38 (20.2%) of whom showed at least one abnormality. We rejected potential donors with factor V Leiden, prothrombin mutation G20210A, and deficiencies in anticoagulant proteins (protein C, protein S, and antithrombin) or coagulation factors. Bleeding and thrombotic events in donors and recipients of the 41 LDLTs were evaluated during 7 days to 70 months follow-up. No major bleeding events were detected in the donors. Neither donor nor recipient experienced venous thrombosis or pulmonary embolism. Among all recipients, six suffered hepatic artery thrombosis including five in the first month probably related to surgery. Deep venous thrombosis and pulmonary embolism are well-known complications of hepatic surgery; Prothrombotic abnormalities in the donor can be transmitted to the recipient, leading to increased risk of serious postoperative events. Although the cost-effectiveness is not definitely established, we recommend systematic screening for hemostatic and prothrombotic disorders to prevent more morbidity of a procedure that already has high risks of bleeding and thrombosis.

AMPD1 genotypes and exercise capacity in McArdle patients.

The purpose of this study was to assess if there exists an association between C34T muscle adenosine monophosphate deaminase ( AMPD1) genotypes (i.e., normal homyzygotes [CC] vs. heterozygotes [ CT]) and directly measured indices of exercise capacity (peak oxygen uptake [VO(2peak)], ventilatory threshold [VT], gross mechanical efficiency [GE], etc.) in 44 Caucasian McArdle patients (23 males, 21 females). All patients performed a graded cycle ergometer test until exhaustion (for VO(2peak) and VT determination) and a 12-min constant-load test at the power output eliciting the VT (for GE determination). We found no significant difference in indices of exercise capacity between CC (n = 18) and CT genotypes (n = 5) in the group of male patients (p > 0.05). In contrast, the VO(2) at the VT was significantly lower (p < 0.05) in CT (n = 4; 7.9 +/- 0.4 ml/kg/min) than in CC female patients (n = 17; 11.0 +/- 0.9 ml/kg/min). In summary, heterozigosity for the C34T allele of the AMPD gene is associated with reduced submaximal aerobic capacity in female patients with McArdle disease and might partly account, in this gender, for the variability that exists in the phenotypic manifestation of the disease.

Sample preparation methods for beeswax characterization by gas chromatography with flame ionization detection.

New and simpler methods of sample preparation to determine several families of compounds in beeswax by conventional and high temperature gas chromatography are proposed. To analyze hydrocarbons and palmitates, a dilution of sample is enough whereas for the total acid content, a hydrolysis and simultaneous methylation with BF3-methanol results more effective than the usual methods; for the total content of alcohols, a further acetylation with acetic anhydride is necessary. Free alcohols are directly acetylated in a sample dissolution but for free acids and monoesterified 1,2,3-propanetriols analysis, a previous extraction with acetonitrile is required. The concentrations of all the compounds studied are expressed in weight percentage referred only to one standard: octadecyl octadecanoate. The precision of the analytical methods has been evaluated showing its importance in the analysis of beeswaxes used in apiculture.

Mobilisation of mesenchymal cells into blood in response to skeletal muscle injury.

Mesenchymal cells recruited to damaged tissues must circulate through the bloodstream. The absolute numbers of circulating mesenchymal stem cells (cMSCs) in two different models of acute and chronic skeletal muscle injury were determined. cMSCs were present in significantly higher numbers in both models than in healthy controls. These results support the hypothesis that MSCs are mobilised into the bloodstream after skeletal muscle tissue damage. These two models (acute and chronic) would be of value in the search for molecular mediators of mobilisation of MSCs into the circulation.

[Current status of clinical nutrition at the network of public hospitals from Castilla y León]. / Situación actual de la nutrición clínica en la red de hospitales públicos de Castilla y León.

INTRODUCTION: Clinical nutrition is an activity realized in most of Health Centres of France, Canada, Great Britain and USA. The aim of our work was to determine activity and resources of Nutrition Units of Hospitals in the Community of Castilla y León. MATERIAL AND METHODS: A questionnaire was send to all Hospitals of Castilla y León (SACYL); Hospital Universitario Rio Hortega, Hospital Clinico Universitario, Hospital Comarcal de Medina del Campo, Hospital General Yagüe-Divino Vallés (Burgos), Complejo Hospitalario de Le6n, Hospital General de Segovia, Hospital Virgen de Sonsoles de Avila, Hospital Virgen de la Concha de Zamora, Hospital Comarcal de Aranda de Duero, Hospital Comarcal de Miranda, Hospital General de Soria, Hospital Clinico Universitario de Salamanca. RESULTS: Nine Centres responded questionnaire (75%). A total of 5 Hospitals had a Unit of Nutrition (55.6%). The results showed an average of 0.37 +/- 0.55 specialists for each 400 beds, 0.87 +/- 0.63 nurses for each 400 beds and 1.91 +/- 2.3 auxiliaries for each 400 beds, with an average of 0.21 +/- 0.41 specialists for each 100,000 habitants, 0.49 +/- 0.36 nurses for each 100,000 habitants and 1.09 +/- 1.2 auxiliaries for each 100,000 habitants. The activity of these Units is demanded by other Units, with an average of 3.2 +/- 3.4 consultations per day. The main diseases of this activity were 33.3% tumoral pathology, 55.6% surgery and 11.1% neurological pathology. Oral supplements were the first intervention tool. Only 3 Centres had a home artificial nutrition consultation. The main diseases of this activity were post surgical patients (33,3%), tumoral pathology (33,3%), neurological pathology (22%) and inflammatory bowel disease (11%). CONCLUSION: Resources in Units of Nutrition of Castilla y Leon were limited. However, activity in Hospital an in home is equal than other areas. New actions of Local Administration are necessaries to follow recommendations of Council of Europe.

Algas epífitas de Bajo Pepito, Isla Mujeres, Quintana Roo, México / Epiphytic algae from Bajo Pepito, Isla Mujeres, Quintana Roo, Mexico

A total of 96 epiphytic algae species were identified from Bajo Pepito, Quintana Roo, México. 60.4% (58) belonged to the Rhodophyta, 19.79% (19) to the Phaeophyta, 16.6% (16) to the Chlorophyta and 3.1% (3) to the Cyanophyta; 49 species (50.5%) were found only in one month, while Heterosiphonia crispella was found in all of the sampled months. That species provided the largest contribution to the biomass of epiphytes. During January we registered the greater biommass and richness of epiphytes species, coincidently with high values of host species cover and rainfall.

Se identificó un total de 96 especies de algas epífitas de Bajo Pepito, Quintana Roo, México; el 60.4% (58) pertenecieron a la división Rhodophyta, 19.79% (19) a la división Phaeophyta, 16.6% (16) a la división Chlorophyta y 3.1% (3) a la división Cyanophyta; 49 especies (51%) se presentaron solamente en un mes de muestreo. Heterosiphonia crispella se presentó en todos los meses de muestreo, y fue la que tuvo mayor contribución en la biomasa de epífitas. En enero se registró la mayor biomasa y riqueza de algas epífitas, lo cual coincidió con valores altos de cobertura de especies hospederas y precipitación pluvial.

Mitochondrial activity is modulated by TNFalpha and IL-1beta in normal human chondrocyte cells.

OBJECTIVE: Pro-inflammatory cytokines play an important role in osteoarthritis (OA). In osteoarthritic cartilage, chondrocytes exhibit an alteration in mitochondrial activity. This study analyzes the effect of tumor necrosis factor-alpha (TNFalpha) and interleukin-1beta (IL-1beta) on the mitochondrial activity of normal human chondrocytes. MATERIALS AND METHODS: Mitochondrial function was evaluated by analyzing the activities of respiratory chain enzyme complexes and citrate synthase, as well as by mitochondrial membrane potential (Deltapsim) and adenosine triphosphate (ATP) synthesis. Bcl-2 family mRNA expression and protein synthesis were analyzed by RNase protection assay (RPA) and Western-blot, respectively. Cell viability was analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and apoptosis by 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) stain. Glycosaminoglycans were quantified in supernatant by a dimethyl-methylene blue binding assay. RESULTS: Compared to basal cells, stimulation with TNFalpha (10 ng/ml) and IL-1beta (5 ng/ml) for 48 h significantly decreased the activity of complex I (TNFalpha=35% and IL-1beta=35%) and the production of ATP (TNFalpha=18% and IL-1beta=19%). Both TNFalpha and IL-1beta caused a definitive time-dependent decrease in the red/green fluorescence ratio in chondrocytes, indicating depolarization of the mitochondria. Both cytokines induced mRNA expression and protein synthesis of the Bcl-2 family. Rotenone, an inhibitor of complex I, caused a significant reduction of the red/green ratio, but it did not reduce the viability of the chondrocytes. Rotenone also increased Bcl-2 mRNA expression and protein synthesis. Finally, rotenone as well as TNFalpha and IL-1beta, reduced the content of proteoglycans in the extracellular matrix of normal cartilage. CONCLUSION: These results show that both TNFalpha and IL-1beta regulate mitochondrial function in human articular chondrocytes. Furthermore, the inhibition of complex I by both cytokines could play a key role in cartilage degradation induced by TNFalpha and IL-1beta. These data could be important for understanding of the OA pathogenesis.

Protein calorie restriction has opposite effects on glucose metabolism and insulin gene expression in fetal and adult rat endocrine pancreas.

We previously demonstrated that fetuses from undernourished pregnant rats exhibited increased beta-cell mass and hyperinsulinemia, whereas keeping food restriction until adult age caused reduced beta-cell mass, hypoinsulinemia, and decreased insulin secretion. Because these alterations can be related to insulin availability, we have now investigated early and long-term effects of protein calorie food restriction on insulin mRNA levels as well as the possible mechanisms that could modulate the endogenous insulin mRNA content. We used fetuses at 21.5 days of gestation proceeding from food-restricted rats during the last week of pregnancy and 70-day-old rats undernourished from day 14 of gestation until adult age and with respective controls. Insulin mRNA levels, glucose transporters, and total glycolysis and mitochondrial oxidative fluxes were evaluated. We additionally analyzed undernutrition effects on signals implicated in glucose-mediated insulin gene expression, especially pancreatic duodenal homeobox-1 (PDX-1), stress-activated protein kinase-2 (p38/SAPK2), and phosphatidylinositol 3-kinase. Undernourished fetuses showed increased insulin mRNA, oxidative glucose metabolism, and p38/SAPK2 levels, whereas undernutrition until adult age provoked a decrease in insulin gene expression, oxidative glucose metabolism, and PDX-1 levels. The results indicate that food restriction caused changes in insulin gene expression and content leading to alterations in glucose-stimulated insulin secretion. The molecular events, increased p38/SAPK2 levels in fetuses and decreased PDX-1 levels in adults, seem to be the responsible for the altered insulin mRNA expression. Moreover, because PDX-1 activation appears to be regulated by glucose-derived metabolite(s), the altered glucose oxidation caused by undernutrition could in some manner affect insulin mRNA expression.