Substrates' and products' inhibition in fructanase production by a new Kluyveromyces marxianus CF15 from Agave tequilana fructan in a batch reactor.

This study focuses on fructanase production in a batch reactor by a new strain isolated from agave juice (K. marxianus var. drosophilarum) employing different Agave tequilana fructan (ATF) concentrations as substrate. The experimental data suggest that the fructanase production may be inhibited or repressed by high substrate (50 g/L) and ethanol (20.7 g/L) concentrations present in culture medium. To further analyze these phenomena an unstructured kinetic mathematical model taking into account substrate and products inhibition was proposed and fitted. The mathematical model considers six reaction kinetics and the ethanol evaporation, and predicts satisfactorily the biomass, fructan, glucose, fructose, ethanol, and fructanase behavior for different raw material initial concentrations. The proposed model is the first to satisfactorily describe the production of fructanase from branched ATF with a new strain of K. marxianus.

Physical performance of elite and subelite Spanish female futsal players.

The aim of this study was to determine the parameters distinguishing top female futsal players from lower level players. Twenty-seven female futsal players participated in the study, composed of professional first division (elite; n = 15) players and semi-professional second division players (sub-elite; n = 14). Active and passive straight leg raise tests, isokinetic strength of the knee extensor and flexor muscles at 60° · s(-1) angular velocity, squat and counter movement jumps, 30 m sprint, 30 m agility, repeated sprint ability test and maximum ball speed during shooting were measured. The elite players were more agile and kicked harder than sub-elite players in maximum ball speed during the shooting test (P ≤ 0.05). However, no significant differences between teams were observed in active and passive hamstring flexibility, jumping ability, repeated sprint ability test, 30 m sprint time, H/Q ratio and absolute and relative torque from 60° · s(-1) angular velocity. Based on these findings we conclude that agility and maximum ball speed during shooting may be important determinant factors of Spanish female futsal actions and could distinguish ability at high-level games.

Production of fructanase by a wild strain of Saccharomyces cerevisiae on tequila agave fructan.

A new wild strain of Saccharomyces cerevisiae (CF3) isolated from tequila must was evaluated for production of fructanase on Agave tequilana Weber fructan (FT). Fructanase activity (F) was assessed by a 3(3) factorial design (substrate, temperature and pH). High enzymatic activity (31.1 U/ml) was found at 30 °C, pH 5, using FT (10 g/l) as substrate. The effect of initial substrate concentration on F (FT0, 5.7-66 g/l) was studied and it was found that F was highest (44.8 U/ml) at FT0 25 g/l. A 2(2) factorial experimental design with five central points was utilized to study the effect of stirring and aeration on fructanase activity; stirring exhibited a stronger effect on F. The ratio fructanase to invertase (F/S) was 0.57, which confirms that the enzymes are fructanase. Crude fructanase reached high substrate hydrolysis (48 wt%) in 10 h. It is shown that S. cerevisiae CF3 was able to produce large amounts of fructanase by growing it on fructan from A. tequilana.

Development of a new oxygen consumption rate assay in cultures of Acanthamoeba (Protozoa: Lobosea) and its application to evaluate viability and amoebicidal activity in vitro.

A new fluorometric method has been developed for measuring the oxygen consumption rate (OCR) of Acanthamoeba cultures in microplates and for screening molecules with amoebicidal activity against this microorganism. The use of a biofunctional matrix (containing an oxygen-sensitive fluorogenic probe) attached to the microplate wells allowed continuous measurement of OCR in the medium, hence assessment of amoebic growth. The new OCR method applied to cell viability yielded a linear relationship and monitoring was much quicker than with indirect viability assays previously used. In addition, two drugs were tested in a cytotoxicity assay monitored by the new OCR viability test. With this procedure, the standard amoebicidal drug chlorhexidine digluconate showed an IC50 of 3.53 + 1.3 mg/l against Acanthamoeba polyphaga and 3.19 + 1.2 mg/l against Acanthamoeba castellanii, whereas a cationic dendrimer [G1Si(NMe3+)4] showed an IC50 of 6.42 + 1.3 mg/l against A. polyphaga. These data agree with previous studies conducted in our laboratory. Therefore, the new OCR method has proven powerful and quick for amoebicidal drug screening and is likely to be applied in biochemical studies concerning protozoa respiration and metabolism.

Pressure pain sensitivity of the scalp in patients with nummular headache: a cartographic study.

Nummular headache (NH) is characterized by focal pain fixed within a small round or elliptical area of the head surface. Sensory dysfunction is apparently restricted to the symptomatic area, but a thorough analysis of cranial pain sensitivity has not been performed. Pressure pain sensitivity maps were constructed for 21 patients with NH and 21 matched healthy controls. In each subject pressure pain thresholds (PPT) were measured on 21 points distributed over the scalp. In each patient PPT were also measured in the symptomatic area and at a non-symptomatic symmetrical point. In both groups an anterior to posterior gradient was found on each side, with no significant differences of PPT measurements between sides or groups. In patients with NH, only the symptomatic area showed a local decrease of PPT (significant in comparison with the non-symptomatic symmetrical point, P < 0.001). These findings further support that NH is a non-generalized disorder with a peripheral source.

Ultrastructural and cytochemical demonstration of peroxisomes in cultured fibroblasts from patients with peroxisomal deficiency disorders.

The oxidation of very long chain fatty acids and synthesis of ether glycerolipids (plasmalogens) occurs mainly in peroxisomes. Zellweger's cerebrohepatorenal syndrome (CHRS) is a rare, inherited metabolic disease characterized by an apparent absence of peroxisomes, an accumulation of very long chain fatty acids, and a decrease of plasmalogens in tissues and cultured fibroblasts from these patients. As peroxisomes are ubiquitous in mammalian cells, we examined normal and CHRS-cultured fibroblasts for their presence, using an electron microscopic histochemical procedure for the subcellular localization of catalase, a peroxisomal marker enzyme. Small (0.08-0.20 micron) round or slightly oval peroxisomes were seen in both normal and CHRS fibroblasts. The number of peroxisomes was analyzed morphometrically and found to be significantly reduced in all CHRS cell lines. These results are discussed in relation to the underlying defect in peroxisomal function and biogenesis in this disease.

Migrainous corpalgia: body pain and allodynia associated with migraine attacks.

Cephalic and extracephalic allodynia are recognized as a common sign of sensory sensitization during migraine episodes. However, the occurrence of body pain in migraine has not been thoroughly explored. Here we report three patients presenting with spontaneous body pain in association with migraine attacks. A 41-year-old woman experienced face and limb pain along with migraine headaches; it started before, during or after headache, was usually ipsilateral to head pain, and could last from minutes to days. A 39-year-old woman had pain in her right limbs, back and neck for 30-60 min prior to right-sided migraine headaches. A 30-year-old woman perceived pain in her left upper limb for 24-48 h prior to left-sided migraine headaches. All patients had allodynia to mechanical stimuli over the painful areas. Spontaneous body pain may be associated with migraine attacks. Together with allodynia, this might be a consequence of central sensitization.

Effects of whole-body vibration training in patients with multiple sclerosis: A systematic review. / Efectos del entrenamiento vibratorio de cuerpo completo en pacientes con esclerosis múltiple: una revisión sistemática.

INTRODUCTION: Multiple sclerosis (MS) is an autoimmune inflammatory disease of the central nervous system. MS is characterised by nerve demyelination that can alter nerve transmission and lead to such symptoms as fatigue, muscle weakness, and impaired motor function. There are 47 000 people with MS in Spain. Vibration training can be an effective and complementary alternative to traditional exercise to treat patients with MS. The aim of this study was to analyse the effectiveness of vibration training programmes in patients with MS. DEVELOPMENT: We searched 5 electronic databases (PubMed, SPORTDiscus, SciELO, Lilacs, IBECS, and ISI Web of Knowledge) in August 2015. By using a set of keywords, we found studies linking vibration training and MS and included randomised controlled trials that applied vibration training to patients with MS. Our search yielded 71 studies. Only 9 of them were included after removing duplicate studies and those which were not relevant according to our selection criteria. These studies obtained different outcomes. CONCLUSIONS: Some studies found improvements in muscle strength, functional capacity, coordination, resistance, balance, and some areas of MSSS-88. However, we identified limitations in some of these studies and there are still few publications on vibration training and multiple sclerosis to ensure training effectiveness.

Bilateral deficit in explosive force related to sit-to-stand performance in older postmenopausal women.

The purpose of this study was to determine whether the bilateral deficit (BLD) for maximal voluntary force (MVF) and rate of force development (RFD) influences sit-to-stand in older postmenopausal women. Fourteen women performed unilateral and bilateral maximal voluntary contractions during isometric leg-extension. The MVF and RFD over consecutive 50ms periods (0-50, 50-100 and 100-150ms) after force onset and the time to sit-to-stand test were calculated. There was only a BLD for RFD 0-50ms and 50-100ms. The time of sit-to-stand was moderately correlated to BLD for RFD 0-50ms (r=0.505; 95% CI: -0.035 to 0.817; P=0.06), but after controlling for physical activity level the relationship was stronger and statistically significant (r=0.605; 95% CI: 0.109 to 0.859; P=0.029). These results suggest that the BLD for explosive force (0-50ms) might represent a performance-limiting factor for sit-to-stand transfer in postmenopausal women and could be dependent of the physical activity level. Trial registered at Clinical Trials Gov.: NCT02434185.

Health-related quality of life of rheumatic disease patients treated in a specialized IPS in Medellin, Colombia.

PURPOSE: To compare Health-Related Quality of Life (HRQOL) profiles and their associated factors in people with rheumatoid arthritis (RA), spondyloarthritis (SpA), fibromyalgia (FM) and rheumatoid comorbidity treated in a specialized health center (SHC) in Medellin, Colombia. METHODS: A cross-sectional analytical study was performed with 93 RA patients, 30 SpA patients, 41 primary FM patients and 48 secondary FM patients with a concurrent diagnosis of RA or SpA. A demographic, socioeconomic, and clinical survey (the IPAQ, International Physical Activity Questionnaire) and the SF-36 survey validated in Medellin were administered. The statistical analyses were executed using SPSS 21.0 Results. Significant differences were found in all HRQOL dimensions among the patients, with physical performance perceived as the worst in the four groups. FM had the worst HRQOL profile, whereas the least affected group was SpA. Patients with RA and rheumatoid comorbidity shared similar HRQOL scores. The years of study, age and economic satisfaction variables were associated with the physical performance, vitality, social functioning, and mental health domains. CONCLUSION: The HRQOL profile was negative in patients with rheumatic diseases and lower in patients with FM. Additionally, variables or subgroups with greater deterioration were identified. This information will be useful for health activities and generate evidence in favor of incorporating HRQOL measurements into rheumatology services to complement clinical evaluations.

Effectiveness of pelvic floor muscle training in treating urinary incontinence in women: A current review. / Efectividad del entrenamiento de la musculatura del suelo pélvico en el tratamiento de la incontinencia urinaria en la mujer: una revisión actual.

OBJECTIVE: To analyse the content of various published studies related to physical exercise and its effects on urinary incontinence and to determine the effectiveness of pelvic floor training programmes. METHOD: We conducted a search in the databases of PubMed, CINAHL, the Cochrane Plus Library, The Cochrane Library, WOS and SPORTDiscus and a manual search in the Google Scholar metasearcher using the search descriptors for documents published in the last 10 years in Spanish or English. The documents needed to have an abstract or complete text on the treatment of urinary incontinence in female athletes and in women in general. RESULTS: We selected 3 full-text articles on treating urinary incontinence in female athletes and 6 full-text articles and 1 abstract on treating urinary incontinence in women in general. The 9 studies included in the review achieved positive results, i.e., there was improvement in the disease in all of the studies. CONCLUSIONS: Physical exercise, specifically pelvic floor muscle training programmes, has positive effects on urinary incontinence. This type of training has been shown to be an effective programme for treating urinary incontinence, especially stress urinary incontinence.

The effects of intermittent hypoxia training on hematological and aerobic performance in triathletes.

The aim of the present research was to analyze modifications on hematological and aerobic performance parameters after a 7-week intermittent hypoxia training (IHT) program. Eighteen male trained triathletes were divided in two groups: an intermittent hypoxia training group (IHTG: n: 9; 26.0 ± 6.7 years; 173.3 ± 5.9 cm; 66.4 ± 5.9 kg; VO2max: 59.5 ± 5.0 ml/kg/min) that conducted a normoxic training plus an IHT and a control group (CG: n: 9; 29.3 ± 6.8 years; 174.9 ± 4.6 cm; 59.7 ± 6.8 kg; VO2max: 58.9 ± 4.5 ml/kg/min) that performed only a normoxic training. Training process was standardized across the two groups. The IHT program consisted of two 60-min sessions per week at intensities over the anaerobic threshold and atmospheric conditions between 14.5 and 15% FiO2. Before and after the 7-week training, aerobic performance in an incremental running test and hematological parameters were analyzed. After this training program, the IHTG showed higher hemoglobin and erythrocytes (p < 0.05) values than in the CG. In terms of physiological and performance variables, between the two groups no changes were found. The addition of an IHT program to normoxic training caused an improvement in hematological parameters but aerobic performance and physiological variables compared to similar training under normoxic conditions did not increase.

Effect of 6 weeks of whole body vibration training on total and segmental body composition in healthy young adults.

The applied use of new technologies to enhance performance and improve health has been increasing. Initially, whole body vibration training (WBVT) was used as system to improve elite athlete performance. However, this is also used to improve body composition, especially there is a great attention on the effectiveness of WBVT to reduce fat and body weight, with a potential increase in muscle tissue. The aim of this study was to investigate the effects of a 6-week vibration-training program on total and segmental body composition in a group of physically healthy participants. The final study sample included 64 healthy young adults. Subjects were randomly allocated into the control group (CG: n = 26; 16 males and 10 females) and the experimental group (EGWBVT: n = 38; 19 males and 19 females). The program lasted six weeks with a frequency of three sessions per week and each session varied in intensity. There were not found statistically significant differences in any of the body composition variables analysed. This study suggests that a six-week vibration-training program with an increasing intensity (7.2 g-32.6 g) in healthy young adults that are not overweight did not alter total and segmental body composition.

Modulation of epinephrine-stimulated gluconeogenesis by insulin in hepatocytes isolated from genetically obese (fa/fa) Zucker rats.

Genetically obese (fa/fa) Zucker rats present an impaired response of hepatic glucose production to the inhibition by insulin. In this work, we have investigated the modulation by this hormone of epinephrine-stimulated gluconeogenesis, in hepatocytes isolated from obese (fa/fa) rats and their lean (Fa/-) littermates. Epinephrine (1 microM) caused a maximal stimulation of [14C]lactate conversion to [14C]glucose in hepatocytes isolated from either obese or lean animals. The stimulation of gluconeogenesis by epinephrine was accompanied by a significant reduction of fructose 2,6-bisphosphate levels, an inactivation of both pyruvate kinase and 6-phosphofructo 2-kinase, and by a 2-fold increase in the cellular concentrations of cAMP. The presence of insulin in the incubation medium antagonized, in a concentration-dependent manner, the effects of epinephrine. In hepatocytes isolated from lean rats, the reversion caused by insulin was complete, the concentration required for half-maximal insulin action ranging from 0.22 to 0.56 nM. In contrast, in obese rat hepatocytes, insulin only partially blocked epinephrine-mediated effects, and the sensitivity to insulin was 2- to 4-fold lower, as indicated by the corresponding half-maximal insulin action values. Furthermore, insulin (10 nM) almost completely blocked the increase in cAMP levels induced by epinephrine in lean rat hepatocytes, whereas it only provoked a small and nonsignificant reduction of epinephrine-stimulated levels of the cyclic nucleotide in hepatocytes obtained from obese rats.

Impairment of the modulation by glucose of hepatic gluconeogenesis in the genetically obese (fa/fa) Zucker rat.

Genetically obese (fa/fa) Zucker rats show oral glucose intolerance, an alteration that has been attributed at least in part to an impaired suppression of hepatic glucose output after the ingestion of glucose. In this work, we studied the influence of different concentrations of glucose (2.5-30 mM) on gluconeogenesis from a mixture of [14C]lactate-pyruvate as well as on fructose 2,6-bisphosphate levels, pyruvate kinase activity, and flux through the reaction catalyzed by 6-phosphofructo-1-kinase, in hepatocytes isolated from fed obese (fa/fa) or lean (Fa/-) rats. In hepatocytes isolated from lean rats, incubation with increasing concentrations of glucose caused a dose-dependent inhibition of gluconeogenesis (5.02 +/- 0.54 and 1.82 +/- 0.33 mumol lactate converted to glucose/g cells.20 min in hepatocytes incubated in the presence of 2.5 and 30 mM glucose, respectively; n = 4 experiments; P < 0.01) together with a significant elevation of the fructose 2,6-bisphosphate content and a stimulation of the flux through 6-phosphofructo-1-kinase reaction. Glucose also provoked a dose-dependent activation of pyruvate kinase in the absence of changes in the cellular concentration of cAMP. In liver cells from obese animals, gluconeogenesis was not significantly modified by raising the glucose concentration in the incubation medium (1.26 +/- 0.11 and 0.83 +/- 0.14 mumol lactate converted to glucose/g cells.20 min in hepatocytes incubated with 2.5 and 30 mM glucose, respectively; n = 4 experiments; P = 0.11) despite significant increases in both fructose 2,6-bisphosphate levels and flux through the 6-phosphofructo-1-kinase reaction. In these cells, pyruvate kinase was only slightly activated by high glucose concentrations. These results indicate that, unlike fructose 2,6-bisphosphate levels and flux through the 6-phosphofructo-1-kinase reaction, hepatic gluconeogenesis is unresponsive to high glucose concentrations in genetically obese (fa/fa) rats.

Decreased responsiveness of basal gluconeogenesis to insulin action in hepatocytes isolated from genetically obese (fa/fa) Zucker rats.

In vivo studies have demonstrated that hepatic glucose production is poorly responsive to insulin in genetically obese Zucker rats. In this work, we have investigated the modulation by insulin of basal gluconeogenesis, fructose 2,6-bisphosphate levels, and pyruvate kinase and 6-phosphofructo 2-kinase activities in hepatocytes isolated from fed obese (fa/fa) or lean (Fa/-) rats. Gluconeogenesis was estimated by the conversion of a mixture of [14C]lactate-pyruvate to [14C]glucose. Basal gluconeogenesis was significantly reduced in hepatocytes isolated from obese rats compared to that measured in hepatocytes from lean animals (0.63 +/- 0.09 vs. 1.47 +/- 0.05 mumol lactate converted to glucose/g cells.20 min; n = 3-4; P < 0.001). In hepatocytes isolated from lean rats, insulin, without affecting the cellular cAMP concentration, caused a dose-dependent inhibition of the rate of gluconeogenesis, which was accompanied by a significant increase in fructose 2,6-bisphosphate levels and activation of both pyruvate kinase and 6-phosphofructo 2-kinase. In contrast, in hepatocytes isolated from obese (fa/fa) rats, neither basal gluconeogenesis nor any of the other metabolic parameters mentioned were significantly modified by insulin, even when assayed at high hormonal concentrations (10 nM). These results demonstrate a lack of responsiveness of hepatic gluconeogenesis to short term insulin action in genetically obese (fa/fa) rats.

Insulin resistance in genetically obese (fa/fa) rats: changes in the glycosyl-phosphatidylinositol signaling system in isolated hepatocytes.

In different types of mammalian cells, insulin has been shown to promote the release of an inositol phosphate glycan (InsP-glycan) through the hydrolysis of a glycosyl-phosphatidylinositol (glycosyl-PtdIns). This InsP-glycan, which has been demonstrated to be taken up by intact cells, may mediate some of the biological effects of insulin. We have investigated how the insulin resistance expressed in genetically obese (fa/fa) rats affects the glycosyl-PtdIns signaling system in isolated hepatocytes compared to what occurs in hepatocytes isolated from lean (Fa/-) rats. The hepatocyte content of glycosyl-PtdIns was reduced by about 30% in obese rats, with respect to that measured in lean rats (2553 +/- 138 vs. 3334 +/- 115 dpm/mg protein; P < 0.01; n = 5). This reduction was accompanied by a marked blockade of the insulin-mediated glycosyl-PtdIns hydrolysis as well as a decrease (approximately 30%) in the rate of InsP-glycan uptake by the isolated liver cells. Obese Zucker rat hepatocytes also showed a significant decrease in the effects of both insulin and InsP-glycan on the stimulation of glycogen synthesis and the activation of glycogen synthase compared to hepatocytes isolated from lean rats. Our results demonstrate that genetic obesity in Zucker (fa/fa) rats is associated with an impairment of the glycosyl-PtdIns-dependent insulin signaling system.

Impairment of glycosyl-phosphatidylinositol-dependent insulin signaling system in isolated rat hepatocytes by streptozotocin-induced diabetes.

The addition to different types of cells of an inositol-phosphate glycan, generated by the phospholipase C-catalyzed hydrolysis of a insulin-sensitive glycosyl-phosphatidylinositol (glycosyl-PI), mimics some of the biological effects of this hormone. Recently, a specific, time-, dose-, and energy-dependent transport system for this inositol-phosphate glycan has been identified in isolated rat hepatocytes. Here, we show that streptozotocin-induced diabetes mellitus reduced (by about 60%) the basal content of the insulin-sensitive glycosyl-PI in isolated rat hepatocytes. Moreover, streptozotocin-induced diabetes blocked the hydrolysis of the glycosyl-PI in response to insulin, diminished inositol phosphate-glycan uptake by the hepatocytes, and abolished the stimulatory effect of this compound on glycogen synthesis. All these metabolic changes caused by streptozotocin administration were reversed by treatment of the animals with insulin. Our results support the hypothesis that insulin resistance in streptozotocin-induced diabetic rats is related to the impairment of glycosyl-PI metabolism.

Effect of adrenalectomy and glucocorticoid treatment on the levels of an insulin-sensitive glycosyl-phosphatidylinositol in isolated rat hepatocytes.

Insulin resistance caused by dexamethasone administration to rats was accompanied by a marked decrease in the hepatocyte content of an insulin-sensitive glycosyl-phosphatidylinositol, as well as by a blockade of its hydrolysis in response to this hormone. In contrast, bilateral adrenalectomy provoked a significant increase of the cellular glycosyl-phosphatidylinositol levels. Under all the assayed metabolic conditions, a close direct correlation was established between the basal content of this compound and the number of insulin receptors present in the isolated hepatocytes.