Effects of heat stress abatement on systemic and mammary inflammation in lactating dairy cows.

To examine the effects of evaporative cooling on systemic and mammary inflammation of lactating dairy cows, 30 multiparous Holstein cows (parity = 2.4, 156 d in milk) were randomly assigned to 1 of 2 treatments: cooling (CL) with fans and misters or not (NC). The experiment was divided into a 10-d baseline when all cows were cooled, followed by a 36-d environmental challenge when cooling was terminated for NC cows. The onset of environmental challenge was considered as d 1. Temperature-humidity index averaged 78.4 during the environmental challenge. Milk yield and dry matter intake (DMI) were recorded daily. Blood and milk samples were collected from a subset of cows (n = 9/treatment) on d -3, 1, 3, 7, 14, and 28 of the experiment to measure cortisol, interleukin 10 (IL10), tumor necrosis factor-α (TNF-α), haptoglobin, and lipopolysaccharide binding protein (LBP). Mammary biopsies were collected from a second subset of cows (n = 6/treatment) on d -9, 2, 10, and 36 to analyze gene expression of cytokines and haptoglobin. A subset of cows (n = 7/treatment) who were not subjected to mammary biopsy collection received a bolus of lipopolysaccharides (LPS) in the left rear quarter on d 30 of the experiment. Blood was sampled from cows and milk samples from the LPS-infused quarter were collected at -4, 0, 3, 6, 12, 24, 48, and 96 h relative to infusion, for analyses of inflammatory products. Deprivation of cooling decreased milk yield and DMI. Compared with CL cows, plasma cortisol concentration of NC cows was higher on d 1 but lower on d 28 of the experiment (cooling × time). Deprivation of cooling did not affect circulating TNF-α, IL10, haptoglobin, or LBP. Compared with CL cows, NC cows tended to have higher milk IL10 concentrations but did not show effects in TNF-α, haptoglobin, or LBP. No differences were observed in mammary tissue gene expression of TNF-α, IL10, and haptoglobin. Milk yield declined after LPS infusion but was not affected by treatment. Compared with CL cows, NC cows had greater milk somatic cell count following intramammary LPS infusion. Non-cooled cows had lower circulating TNF-α and IL10 concentrations and tended to have lower circulating haptoglobin concentrations than CL cows. Milk IL10 and TNF-⍺ concentrations were higher 3 h after LPS infusion for NC cows compared with CL cows. Additionally, NC cows tended to have higher milk haptoglobin concentration after LPS infusion than CL cows. In conclusion, deprivation of evaporative cooling had minimal effects on lactating cows' basal inflammatory status, but upregulated mammary inflammatory responses after intramammary LPS infusion.

Effects of evaporative cooling and dietary zinc source on heat shock responses and mammary gland development in lactating dairy cows during summer.

The objective of this study was to examine the effects of evaporative cooling and dietary supplemental Zn source on heat shock responses and mammary gland development of lactating dairy cows during summer. Seventy-two multiparous lactating Holstein cows were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement. Cows were either cooled (CL) or not cooled (NC) and fed diets supplemented with 75 mg of Zn/kg of dry matter (DM) from Zn hydroxychloride (IOZ) or 35 mg of Zn/kg of DM from Zn hydroxychloride plus 40 mg of Zn/kg of DM from Zn-Met complex (ZMC). The 168-d trial included a 12-wk baseline phase when all cows were cooled and fed respective dietary treatments, and a subsequent 12-wk environmental challenge phase when NC cows were deprived of evaporative cooling. Plasma was collected from a subset of cows (n = 24) at 1, 3, 5, 12, 26, 41, 54, 68, 81 d of the environmental challenge to measure heat shock protein (HSP) 70 concentration. Mammary biopsies were collected from another subset of cows (n = 30) at enrollment (baseline samples) and at d 7 and 56 of the environmental challenge to analyze gene expression related to heat shock response, apoptosis and anti-oxidative enzymes, and to examine apoptosis and cell proliferation using immunohistochemistry. Supplemental Zn source did not affect milk yield but NC cows produced less milk than CL cows. Supplemental Zn source had no effect on mammary gene expression of HSP27, 70, and 90 or plasma concentrations of HSP70. The NC cows had greater mammary gene expression of HSP than CL cows. Circulating HSP70 of NC cows gradually increased and was higher at 81 d of environmental challenge compared with CL cows. Relative to IOZ, ZMC cows tended to have lower total mammary cell proliferation but greater mammary apoptosis. There was a tendency of greater TNFRSF1A mRNA expression for ZMC compared with IOZ cows, which may suggest upregulated extrinsic apoptosis. At d 7 of environmental challenge, NC cows had numerically higher mammary apoptosis than CL cows although not statistically significant. The NC cows tended to have greater mRNA expression of CAT and SOD3 regardless of time, and had greater mRNA expression of GPX1 at d 56 and FAS at d 7 of the environmental challenge than CL cows. Relative to CL cows, mammary cell proliferation rate was higher for NC cows at d 56 of the environmental challenge. In conclusion, dietary source of supplemental Zn has substantial effect on mammary cell turnover in lactating dairy cows, and prolonged exposure to heat stress increases mammary cell proliferation.

Effects of milk replacer feeding levels on performance and metabolism of preweaned dairy calves during summer.

The objective of this study was to evaluate the effect of milk replacer (MR) feeding programs on performance and metabolism during summer. At 3 d of age (DOA), calves were randomly assigned to 1 of 4 dietary treatments: control [CON; 0.55 kg dry matter (DM) of a 20% crude protein (CP) and 20% fat MR per day], intermediate (IL; 0.66 kg DM of a 26% CP and 17% fat MR per day), high (HL; 0.77 kg DM of a 26% CP and 17% fat MR per day), or aggressive (AL; 0.87 kg DM of a 26% CP and 17% fat MR per day). Calves were managed similarly and housed in individual polyethylene hutches using sand as a bedding material. Because 3 calves fed the AL diet developed abomasum bloating during the first 30 DOA, the AL treatment was terminated. Milk replacer (12.5% solids) was offered twice daily until 42 DOA, when MR was fed once daily to reduce its intake by 50%. Calves were weaned at 49 DOA and remained in hutches until 56 DOA. Calf starter and water were offered ad libitum. Ambient temperature and relative humidity in and outside the hutches were assessed hourly. Starter and MR intakes were recorded daily. Respiration rate and rectal temperature were determined 3 times each week. Body weight was measured at 3, 14, 28, 42, and 56 DOA. Plasma was collected at 5, 10, 14, 28, 42, 43, 45, 47, 49, 51, and 56 DOA for analysis of glucose, ß-hydroxybutyrate, triglycerides, nonesterified fatty acids, urea nitrogen, and insulin concentrations. There were no treatment effects on starter intake, rectal temperature, or respiration rate. By 7 DOA, calves fed the IL and HL diets consumed the same amount of MR and a higher amount of MR than the CON calves. At wk 2, calves from all treatments had similar MR consumption before returning to the projected intake by design at wk 4. Calves fed the IL and HL treatments had similar body weight but were heavier than those fed the CON diet at wk 6, 7, and 8. Calves fed the IL and HL diets had similar average daily gain, which was higher than that of calves fed the CON diet. There was no difference in plasma metabolites among treatments, but insulin concentration increased as milk allowance increased. In summary, feeding an intermediate level of MR during summer improved calf growth compared with the CON diet, but a higher MR allowance did not support further improvements in calf performance.

Prevalence of Spasticity and Postural Patterns in the Upper Extremity Post Stroke.

INTRODUCTION: A high number of patients with stroke develop upper extremity spasticity, causing abnormal postures and patterns. These alterations limit the use of arm in functional activities and affect social participation. AIM: To determine the prevalence of spasticity and postural patterns of the upper extremity post stroke. MATERIALS AND METHODS: A cross-sectional descriptive design was used with a prospective follow-up. The sample included 136 patients. The study included 3 measuring times; at 10 days (T1), applying a record with sociodemographic-clinical data, the evaluation of muscle tone in the elbow and wrist and the postural patterns of the UE, and at 3 months (T2) and 12 months (T3) post stroke, re-evaluating tone and patterns. Prevalence was calculated through the one-sample chi-squared (χ2) test followed by inspection of the standardized residuals (z) in each cell. The Kappa coefficient evaluated the degree of agreement in elbow and wrist tone. RESULTS: The prevalence of spasticity in the elbow was 37.5% at T1, 57.4% at T2, and 57.4% at T3. At each time there was a high degree of agreement between elbow and wrist tone. Patients developed increased elbow tone between T1 and T2, with maintained tone between T2 and T3. Postural pattern III was the most prevalent according to Hefter's classification. CONCLUSION: The prevalence of spasticity in the elbow and wrist increases between 10 days and 3 months post stroke, and is maintained between 3 and 12 months. The onset of spasticity occurs in almost half of patients during the first 10 days post stroke. Postural pattern III according to Hefter's classification presented the greatest prevalence in the spastic UE.

Impact of Physiological Signals Acquisition in the Emotional Support Provided in Learning Scenarios.

Physiological sensors can be used to detect changes in the emotional state of users with affective computing. This has lately been applied in the educational domain, aimed to better support learners during the learning process. For this purpose, we have developed the AICARP (Ambient Intelligence Context-aware Affective Recommender Platform) infrastructure, which detects changes in the emotional state of the user and provides personalized multisensorial support to help manage the emotional state by taking advantage of ambient intelligence features. We have developed a third version of this infrastructure, AICARP.V3, which addresses several problems detected in the data acquisition stage of the second version, (i.e., intrusion of the pulse sensor, poor resolution and low signal to noise ratio in the galvanic skin response sensor and slow response time of the temperature sensor) and extends the capabilities to integrate new actuators. This improved incorporates a new acquisition platform (shield) called PhyAS (Physiological Acquisition Shield), which reduces the number of control units to only one, and supports both gathering physiological signals with better precision and delivering multisensory feedback with more flexibility, by means of new actuators that can be added/discarded on top of just that single shield. The improvements in the quality of the acquired signals allow better recognition of the emotional states. Thereof, AICARP.V3 gives a more accurate personalized emotional support to the user, based on a rule-based approach that triggers multisensorial feedback, if necessary. This represents progress in solving an open problem: develop systems that perform as effectively as a human expert in a complex task such as the recognition of emotional states.

Experimental study of the susceptibility of a European Aedes albopictus strain to dengue virus under a simulated Mediterranean temperature regime.

Dengue virus (DENV) has re-emerged in Europe driven by the geographic expansion of the mosquito species Aedes albopictus and Aedes aegypti (Diptera: Culicidae) and the introduction of the virus by viraemic travellers. In the present study, the vector competence (VC) of Ae. albopictus collected in Catalonia (northeast Spain) was evaluated for two different DENV strains, DENV-1 and DENV-2, the serotypes responsible for all outbreaks of dengue that have occurred in Europe. Mosquitoes were reared under environmental conditions mimicking the mean temperature and humidity recorded in July on the Mediterranean coast of Catalonia. Mosquitoes were fed on an artificial infectious bloodmeal and, at 14 days post-exposure, infection, disseminated infection and transmission rates (IR, DIR, TR) and transmission efficiency (TE) were determined by testing the virus in the body, legs and saliva. The tested Ae. albopictus strain was found to be susceptible to both DENV-1 and DENV-2 and to be able to transmit DENV-1. This is the first time that the VC of Ae. albopictus for DENV has been tested in Europe in this specific context (i.e. mimicking the Mediterranean temperature and humidity recorded in Catalonia in July). This study confirms the potential of Ae. albopictus to start autochthonous DENV transmission cycles in the Mediterranean basin.

[Rate of readmissions of patients with heart failure: effects of multifactorial educational interventions]. / Efectividad de las intervenciones educativas con enfoque multifactorial y seguimiento vía telefónica sobre la rehospitalización en pacientes con insuficiencia cardiaca: revisión sistemática y meta-análisis.

We performed a systematic review of the literature to analyze the effect on readmission rates of multifactorial educational interventions at the moment of discharge from the hospital and telephone follow up for patients with heart failure. The quality of reports was analyzed using the CONSORT verification list and the GRADE guide. Using Rev Manager 5.3, the relative risk heterogeneity (RR) of readmission was assessed using I2 and Q statistics, and those considered as homogeneous were combined into a single RR as the confidence interval. We included four reports in the review and three were meta-analyzed. The RR for readmission was 1.01 (95% confidence interval (CI): 0.73-1.40; p = 0.96) with educational interventions and telephone follow-up among patients who visited an emergency room (ER). When studies that included readmission (without ER visit) were combined, the RR was 0.83 (95% CI:0.58-1.17). When studies with visits to ER and readmissions were combined, the RR was 0.91 (95% CI:0.73-1.14). We conclude that educational interventions did not have a significant effect on the rate of readmissions of patients with heart failure.

Epidemiological and pathological investigation of fowl aviadenovirus serotypes 8b and 11 isolated from chickens with inclusion body hepatitis in Spain (2011-2013).

Inclusion body hepatitis caused by different fowl aviadenovirus (FAdV) serotypes has been described in several countries in recent years. In Spain, from the spring of 2011 to 2013, an increased number of outbreaks in broiler and broiler breeder flocks from different regions occurred. The objectives of the present work were to carry out the molecular characterization of FAdV strains from Spanish inclusion body hepatitis cases and to study the pathogenicity and viral dynamics of these strains in specific pathogen-free (SPF) chickens. A total of 52 inclusion body hepatitis clinical cases, including 45 from broiler farms and seven from broiler breeder farms, were analysed by conventional polymerase chain reaction and sequencing targeting the FAdV hexon gene. From these, 37 strains were classified as FAdV type 8b, while the remaining 15 were classified as FAdV types 11 (n = 10), 2 (n = 4) and 8a (n = 1). In addition, two different FAdVs belonging to the genotypes 8b and 11 were used for experimental infection. Specific pathogen-free five-day-old birds were inoculated intramuscularly with a high (106.5 tissue culture infective dose (TCID)50/ml) or low (104 TCID50/ml) dose of the above-mentioned FAdVs. No mortality was observed in any of the experimental groups, and only one bird showed evident clinical signs. However, macroscopic and microscopic hepatic lesions, as well as viral DNA, were detected in birds from all infection groups. Inclusion bodies and viral DNA were also detected in the pancreas and in the small and the large intestine in some birds. Long-lasting shedding and transmission to contact birds were confirmed in all infected groups.

Rift Valley fever virus and European mosquitoes: vector competence of Culex pipiens and Stegomyia albopicta (= Aedes albopictus).

Rift Valley fever (RVF) is a mosquito-borne disease caused by the Rift Valley fever virus (RVFV). Rift Valley fever affects a large number of species, including human, and has severe impact on public health and the economy, especially in African countries. The present study examined the vector competence of three different European mosquito species, Culex pipiens (Linnaeus, 1758) form molestus (Diptera: Culicidae), Culex pipiens hybrid form and Stegomyia albopicta (= Aedes albopictus) (Skuse, 1894) (Diptera: Culicidae). Mosquitoes were artificially fed with blood containing RVFV. Infection, disseminated infection and transmission efficiency were evaluated. This is the first study to assess the transmission efficiency of European mosquito species using a virulent RVFV strain. The virus disseminated in Cx. pipiens hybrid form and in S. albopicta. Moreover, infectious viral particles were isolated from saliva of both species, showing their RVFV transmission capacity. The presence of competent Cx. pipiens and S. albopicta in Spain indicates that an autochthonous outbreak of RVF may occur if the virus is introduced. These findings provide information that will help health authorities to set up efficient entomological surveillance and RVFV vector control programmes.

Functional characterization of two new STAT3 mutations associated with hyper-IgE syndrome in a Mexican cohort.

Hyper-IgE syndrome (HIES) is an immunodeficiency disorder that is characterized by distinctive immunologic and non-immunologic manifestations. Although mutations in signal transducer and activator of transcription 3 (STAT3) have been associated with HIES, the exact nature of the relationship is unknown. Here, we characterized the functional activity of STAT3 and its mutations in 11 Mexican patients with autosomal dominant HIES. STAT3 phosphorylation was evaluated by flow cytometry, and in silico analyses were performed to estimate the impact of allelic mutations on the DNA binding and SH2 domains of the STAT3 protein. Electrophoretic mobility shift assays were used to assess whether the STAT3 mutants could bind to the consensus oligonucleotide target in vitro. Two novel mutations [g.58891A>T (Asn395Tyr) and g.59078A>T (Asn425Tyr)] as well as one possible somatic mosaicism were found in several of the patients who bore some remarkable features. However, there were no direct correlations between genotypes and HIES clinical features. STAT3 phosphorylation was found to be lower in the patient cohort than in healthy controls. Moreover, the mutated STAT3 proteins could bind to the Sp1, but not to the STAT3, consensus sequence. From these functional studies, the STAT3 mutations found in our patient cohort were concluded to be deleterious for normal STAT3 function.

Culex pipiens and Stegomyia albopicta (= Aedes albopictus) populations as vectors for lineage 1 and 2 West Nile virus in Europe.

The emerging disease West Nile fever is caused by West Nile virus (WNV), one of the most widespread arboviruses. This study represents the first test of the vectorial competence of European Culex pipiens Linnaeus 1758 and Stegomyia albopicta (= Aedes albopictus) (both: Diptera: Culicidae) populations for lineage 1 and 2 WNV isolated in Europe. Culex pipiens and S. albopicta populations were susceptible to WNV infection, had disseminated infection, and were capable of transmitting both WNV lineages. This is the first WNV competence assay to maintain mosquito specimens under environmental conditions mimicking the field (day/night) conditions associated with the period of maximum expected WNV activity. The importance of environmental conditions is discussed and the issue of how previous experiments conducted in fixed high temperatures may have overestimated WNV vector competence results with respect to natural environmental conditions is analysed. The information presented should be useful to policymakers and public health authorities for establishing effective WNV surveillance and vector control programmes. This would improve preparedness to prevent future outbreaks.

Effects of Steroid Hormone in Avian Follicles.

The aim of the present study was to examine the effects of testosterone (T) and estradiol-17ß (E2) on the production of progesterone (P4) by granulosa cells, and of the E2 on the production of P4 and T by theca internal cells. In the first experiment, granulosa cells isolated from the largest (F1) and third largest (F3) preovulatory follicle were incubated for 4 h in short-term culture system, P4 production by granulosa cells of both F1 and F3 was increased in a dose-dependent manner by ovine luteinizing hormone (oLH), but not T or E2. In the second experiment, F1 and F3 granulosa cells cultured for 48 h in the developed monolayer culture system were recultured for an additional 48 h with increasing doses of various physiological active substances existing in the ovary, including T and E2. Basal P4 production for 48 h during 48 to 96 h of the cultured was about nine fold greater by F1 granulosa cells than by F3 granulosa cells. In substances examined oLH, chicken vasoactive intestinal polypeptide (cVIP) and T, but not E2, stimulated in a dose-dependent manner P4 production in both F1 and F3 granulosa cells. In addition, when the time course of P4 production by F1 granulosa cells in response to oLH, cVIP, T and E2 was examined for 48 h during 48 to 96 h of culture, although E2 had no effect on P4 production by granulosa cells of F1 during the period from 48 to 96 h of culture, P4 production with oLH was found to be increased at 4 h of the culture, with a maximal 9.14 fold level at 6 h. By contrast, P4 production with cVIP and T increased significantly (p<0.05) from 8 and 12 h of the culture, respectively, with maximal 6.50 fold response at 12 h and 6, 48 fold responses at 36 h. Furthermore, when F1 granulosa cells were precultured with E2 for various times before 4 h culture with oLH at 96 h of culture, the increase in P4 production in response to oLH with a dose-related manner was only found at a pretreatment time of more than 12 h. In the third experiment, theca internal cells of F1, F2 and the largest third to fifth preovulatory follicles (F3-5) were incubated for 4 h in short-term culture system with increasing doses of E2. The production of P4 and T by theca internal cells were increased with the addition of E2 of 10(-6) M. These increases were greater in smaller follicles. These results indicate that, in granulosa cells of the hen, T may have a direct stimulatory action in the long term on P4 production, and on E2 in long-term action which may enhance the sensitivity to LH for P4 production, and thus, in theca internal cells, E2 in short term action may stimulate the production of P4 and T.

Characteristics and outcomes of patients admitted to Spanish ICU: A prospective observational study from the ENVIN-HELICS registry (2006-2011).

OBJECTIVE: To describe the case-mix of patients admitted to intensive care units (ICUs) in Spain during the period 2006-2011 and to assess changes in ICU mortality according to severity level. DESIGN: Secondary analysis of data obtained from the ENVN-HELICS registry. Observational prospective study. SETTING: Spanish ICU. PATIENTS: Patients admitted for over 24h. INTERVENTIONS: None. VARIABLES: Data for each of the participating hospitals and ICUs were recorded, as well as data that allowed to knowing the case-mix and the individual outcome of each patient. The study period was divided into two intervals, from 2006 to 2008 (period 1) and from 2009 to 2011 (period 2). Multilevel and multivariate models were used for the analysis of mortality and were performed in each stratum of severity level. RESULTS: The study population included 142,859 patients admitted to 188 adult ICUs. There was an increase in the mean age of the patients and in the percentage of patients >79 years (11.2% vs. 12.7%, P<0.001). Also, the mean APACHE II score increased from 14.35±8.29 to 14.72±8.43 (P<0.001). The crude overall intra-UCI mortality remained unchanged (11.4%) but adjusted mortality rate in patients with APACHE II score between 11 and 25 decreased modestly in recent years (12.3% vs. 11.6%, odds ratio=0.931, 95% CI 0.883-0.982; P=0.008). CONCLUSION: This study provides observational longitudinal data on case-mix of patients admitted to Spanish ICUs. A slight reduction in ICU mortality rate was observed among patients with intermediate severity level.

[Comparative analysis of patients admitted to Spanish Intensive Care Units due to medical and surgical disease]. / Análisis comparativo de pacientes ingresados en Unidades de Cuidados Intensivos españolas por causa médica y quirúrgica.

OBJECTIVE: To describe the characteristics of the patients case-mix admitted to ICUs due to medical and surgical disease, and to compare both groups. DESIGN: Analysis of data covering the period 2006-2011 in the ENVIN-HELICS registry. An observational, prospective, multicenter and voluntary participation study. SETTING: A total of 188 Spanish ICUs. PATIENTS: All patients admitted for more than 24 hours. MAIN VARIABLES: Demographic data, cause of admission, severity scores, length of stay, mortality. RESULTS: A total of 138,999 patients were analyzed. Of these, 65,467 (47.1%) were admitted due to a non-coronary medical cause, 27,785 (20,0%) due to coronary-related illness, 28,044 (20,2%) after elective surgery and 17,613 (12.7%) after urgent surgery. Use of devices, nosocomial infections and isolation of multirresistant organisms were more prevalent in urgent surgery patients. Longer length of stay (median 5 days; interquartile range 2-11) as well as higher severity scale values (APACHE II and SAPS II) corresponded to this same group of patients. Mortality was higher in non-coronay medical patients. On categorizing the patients according to the APACHE II score, mortality was seen to be higher in urgent surgery cases than in elective surgery patients in all groups. The largest difference was observed in the APACHE II score 6-10 group (3% vs. 0.9%) (OR: 2.14, 95% CI 1.825-2.513; p<0.001). CONCLUSIONS: The mortality rate is higher in non-coronary medical patients, though resource use per patient is greater in the urgent surgery cases. The APACHE II scale underestimates mortality in emergency surgery patients.

[Impact of the Interdisciplinary Chronic Pain Rehabilitation Programme in patients with and without sleep disorders]. / Impacto del Programa de Rehabilitación Interdisciplinario de Dolor Crónico en pacientes sin y con trastornos del sueño.

INTRODUCTION: Sleep disorders and chronic pain are linked to each other bidirectionally. Both are related to affective disorders, fatigue, depression, anxiety and drug abuse, and have a significant effect on quality of life. The Interdisciplinary Pain Programme (IDP) aims to relieve the patients' pain and improve their functionality by incorporating healthy postural, sleep and nutritional habits, relaxation techniques, physical exercise and cognitive-behavioural mechanisms. PATIENTS AND METHODS: A retrospective, observational, cross-sectional study was conducted. A total of 323 patients with chronic pain who completed the IDP were examined. They were assessed at the beginning and at the end of the programme with pain, depression, quality of life and insomnia scales, and were then compared between groups with and without insomnia, that is, with an insomnia severity index (ISI) less than 15 versus greater than or equal to 15. Fifty-eight patients were studied by means of polysomnography. RESULTS: A significant improvement (p < 0.0001) in pain, depression and quality of life, as assessed by the visual analogue scale (VAS), the Beck inventory and the Short Form-36 (SF-36) questionnaire was observed in chronic pain patients with an ISI below 15 and in those with an ISI greater than or equal to 15. The results were superior in the group of patients with insomnia. The presence of a high apnoea and hypopnoea index and periodic lower limb movements in patients was not related to improvements on the Beck, SF-36, ISI and VAS scales. CONCLUSIONS: In conclusion, IDP benefits patients with chronic non-cancer-induced pain in several affected areas, in addition to pain, due to a comprehensive treatment. Polysomnography can help diagnose specific pathologies and individualise pharmacological treatment.

TITLE: Impacto del Programa de Rehabilitación Interdisciplinario de Dolor Crónico en pacientes sin y con trastornos del sueño.Introducción. Los trastornos del sueño y el dolor crónico están relacionados bidireccionalmente. Ambos están relacionados con trastornos afectivos, fatiga, depresión, ansiedad y abuso de fármacos, y afectan significativamente a la calidad de vida. El objetivo del Programa Interdisciplinario de Dolor (PRID) es aliviar el dolor del paciente y mejorar su funcionalidad a través de la incorporación de hábitos posturales, del sueño y nutricionales saludables, técnicas de relajación, ejercicio físico y mecanismos cognitivoconductuales. Pacientes y métodos. Se realizó un estudio retrospectivo, observacional y transversal. Se examinó a 323 pacientes con dolor crónico que completaron el PRID. Se les evaluó al principio y al final del programa con escalas de dolor, depresión, calidad de vida e insomnio, y se les comparó entre grupos con y sin insomnio ­índice de gravedad del insomnio (ISI) menor de 15 frente a mayor o igual a 15­. Se estudió a 58 pacientes con polisomnografía. Resultados. Se observó una mejoría significativa (p < 0,0001) del dolor, la depresión y la calidad de vida evaluados mediante la escala analógica visual (EVA), el inventario de Beck y el cuestionario Short Form-36 (SF-36), tanto en pacientes con dolor crónico con ISI menor de 15 como ISI mayor o igual a 15. Los resultados fueron superiores en el grupo de pacientes con insomnio. La presencia de un índice de apneas e hipopneas elevado y movimientos periódicos de los miembros inferiores en los pacientes no se relacionó con la mejoría de las escalas de Beck, SF-36, ISI y EVA. Conclusiones. En conclusión, el PRID beneficia a los pacientes con dolor crónico no oncológico en varias esferas afectadas, además del dolor, mediante un tratamiento integral. La polisomnografía puede ayudar a diagnosticar patologías específicas e individualizar el tratamiento farmacológico.

Prevalence of digestive disorders associated with imported Chagas disease (PADChI study): an observational study.

BACKGROUND: Chagas disease (CD) is a parasitic disease caused by Trypanosoma cruzi, in which up to 10-20% of those affected may suffer digestive disorders. Multiple studies have been carried out on CD in non-endemic countries, mainly related to cardiological involvement. However, digestive disorders have not been analyzed in such depth. The objective of the study was to determine the prevalence of digestive disorders in imported CD at the time of first care. METHODS: An observational cross-sectional descriptive analysis of imported CD was performed. Chagasic structural damage and infectious digestive comorbidity were evaluated. The association between Chagasic structural damage and heart disease in Chagas patients was also investigated. RESULTS: After reviewing a total of 1,216 medical records, those of 464 patients were selected for analysis. Globally, the prevalence of digestive disorders in imported Chagas was 57.76%, 95% CI (53.25-62.27). The prevalence of comorbidity of infectious diseases was 40.73% CI 95% (36.25-45.22). Colonic abnormalities were found in 84 of 378 barium enema patients. CD-related esophageal abnormalities were present in 63 of 380 patients studied with esophagogram. CONCLUSIONS: The prevalence of digestive disorders associated with CD is high, so the presence of infectious diseases (mainly parasitic and H. pylori infection) should be ruled out. It is important to exclude structural involvement in all symptomatic patients, and asymptomatic patients should also be considered and offered.

Role of Rhizobium endoglucanase CelC2 in cellulose biosynthesis and biofilm formation on plant roots and abiotic surfaces.

BACKGROUND: The synthesis of cellulose is among the most important but poorly understood biochemical processes, especially in bacteria, due to its complexity and high degree of regulation. In this study, we analyzed both the production of cellulose by all known members of the Rhizobiaceae and the diversity of Rhizobium celABC operon predicted to be involved in cellulose biosynthesis. We also investigated the involvement in cellulose production and biofilm formation of celC gene encoding an endoglucanase (CelC2) that is required for canonical symbiotic root hair infection by Rhizobium leguminosarum bv. trifolii. RESULTS: ANU843 celC mutants lacking (ANU843ΔC2) or overproducing cellulase (ANU843C2+) produced greatly increased or reduced amounts of external cellulose micro fibrils, respectively. Calcofluor-stained cellulose micro fibrils were considerably longer when formed by ANU843ΔC2 bacteria rather than by the wild-type strain, in correlation with a significant increase in their flocculation in batch culture. In contrast, neither calcofluor-stained extracellular micro fibrils nor flocculation was detectable in ANU843C2+ cells. To clarify the role of cellulose synthesis in Rhizobium cell aggregation and attachment, we analyzed the ability of these mutants to produce biofilms on different surfaces. Alteration of wild-type CelC2 levels resulted in a reduced ability of bacteria to form biofilms both in abiotic surfaces and in planta. CONCLUSIONS: Our results support a key role of the CelC2 cellulase in cellulose biosynthesis by modulating the length of the cellulose fibrils that mediate firm adhesion among Rhizobium bacteria leading to biofilm formation. Rhizobium cellulose is an essential component of the biofilm polysaccharidic matrix architecture and either an excess or a defect of this "building material" seem to collapse the biofilm structure. These results position cellulose hydrolytic enzymes as excellent anti-biofilm candidates.

A functional study of nucleocytoplasmic transport signals of the EhNCABP166 protein from Entamoeba histolytica.

EhNCABP166 is an Entamoeba histolytica actin-binding protein that localizes to the nucleus and cytoplasm. Bioinformatic analysis of the EhNCABP166 amino acid sequence shows the presence of 3 bipartite nuclear localization signals (NLS) and a nuclear export signal (NES). The present study aimed to investigate the functionality of these signals in 3 ways. First, we fused each potential NLS to a cytoplasmic domain of ehFLN to determine whether the localization of this domain could be altered by the presence of the NLSs. Furthermore, the localization of each domain of EhNCABP166 was determined. Similarly, we generated mutations in the first block of bipartite signals from the domains that contained these signals. Additionally, we added an NES to 2 constructs that were then evaluated. We confirmed the intranuclear localization of EhNCABP166 using transmission electron microscopy. Fusion of each NLS resulted in shuttling of the cytoplasmic domain to the nucleus. With the exception of 2 domains, all of the evaluated domains localized within the nucleus. A mutation in the first block of bipartite signals affected the localization of the domains containing an NLS. The addition of an NES shifted the localization of these domains to the cytoplasm. The results presented here establish EhNCABP166 as a protein containing functional nuclear localization signals and a nuclear export signal.

Identification and evaluation of inhibitors of the EhGEF1 protein from Entamoeba histolytica.

The Dbl family of guanine nucleotide exchange factors (GEFs) is made up of a vast array of members that participate in the activation of the Rho family of small GTPases. Dbl-family proteins promote the exchange of guanosine diphosphate/guanosine triphosphate (GDP/GTP) in their target molecules, resulting in the activation of a variety of signaling pathways involved in diverse cellular events, such as actin-cytoskeleton remodeling, cellular invasion, cell movement, and other functions. It has been reported that members of the Dbl family have important roles in several cellular events in Entamoeba histolytica. These include activation of the actin cytoskeleton, cytokinesis, capping, uroid formation, cellular proliferation, erythrophagocytosis, cell migration, and chemotaxis. Here, we report the identification and testing of inhibitors of the E. histolytica guanine nucleotide exchange factor 1 (EhGEF1) protein (the research compounds 2BYRF, 2BY05, 2BYT6, 2BYLX, and 2BYPD), which decreased the in vitro ability of the protein to exchange GDP/GTP at its target GTPases, EhRacG and EhRho1, by 14.9-85.2%. Interestingly, the drug 1,1'-(1,2-phenylene)-bis-(1H-pyrrole-2,5-dione), which completely inhibits the GEF activity of the Trio protein in human cells, decreases the GEF activity of the EhGEF1 protein on the EhRacG and EhRho1 GTPases by 55.7% and 3.2%, respectively. The identification and evaluation of such inhibitors opens up the possibility of obtaining a new pharmacological tool to study the function of amoeba GEF proteins, their roles in various Rho GTPase-mediated signaling pathways, and the repercussions of modulating their activities with respect to several mechanisms related to E. histolytica pathogenesis.

Characterization of root-nodulating bacteria associated to Prosopis farcta growing in the arid regions of Tunisia.

Diversity of 50 bacterial isolates recovered from root nodules of Prosopis farcta grown in different arid soils in Tunisia, was investigated. Characterization of isolates was assessed using a polyphasic approach including phenotypic characteristics, 16S rRNA gene PCR--RFLP and sequencing, nodA gene sequencing and MLSA. It was found that most of isolates are tolerant to high temperature (40°C) and salinity (3%). Genetic characterization emphasizes that isolates were assigned to the genus Ensifer (80%), Mesorhizobium (4%) and non-nodulating endophytic bacteria (16%). Forty isolates belonging to the genus Ensifer were affiliated to Ensifer meliloti, Ensifer xinjiangense/Ensifer fredii and Ensifer numidicus species. Two isolates belonged to the genus Mesorhizobium. Eight isolates failing to renodulate their host plant were endophytic bacteria and belonged to Bacillus, Paenibacillus and Acinetobacter genera. Symbiotic properties of nodulating isolates showed a diversity in their capacity to infect their host plant and fix atmospheric nitrogen. Isolate PG29 identified as Ensifer meliloti was the most effective one. Ability of Prosopis farcta to establish symbiosis with rhizobial species confers an important advantage for this species to be used in reforestation programs. This study offered the first systematic information about the diversity of microsymbionts nodulating Prosopis farcta in the arid regions of Tunisia.