Isolation and Selection of Streptomyces Species from Semi-arid Agricultural Soils and Their Potential as Producers of Xylanases and Cellulases.

The Mezquital Valley (MV), Mexico, is a semi-arid region whose main economic activity is agriculture, this zone is characterized by the use of wastewater for crop irrigation. This condition has increased the amount nutrients in soils, organic carbon content and native microorganisms. The Streptomyces species are a group of saprophytic bacteria that represent between 20 and 60% of the total microbial population in soils, capable of producing metabolites of commercial importance. In this work, Streptomyces species were isolated from agricultural soils of the MV and was evaluated the production of endoglucanases (CMCase) and xylanases (Xyl) in Solid-State Cultivation (SSC). From soil samples, 73 possible strains of Streptomyces species were isolated for their ability to produce CMCase and Xyl in SSC. The study also included its characterization by morphological characteristics. Of the isolated microorganisms, 38 strains were selected as strong enzyme producers according to the measurement of the halo generated in plate and by growth on barley straw as only carbon source. Two different sizes of barley straw particle were tested, finding that the greatest enzymatic activity was observed in particle size 12. Three strains of Streptomyces species were chosen which presented the best catalytic capacities, a maximum of 100.69 AU Xyl/gram dry matter (gdm), 82 AU Xyl/gdm and 26.02 AU CMCase/gdm for strains 30, 28 and 12, respectively. The strains were identified by ribosomal gen16s sequence and identified as S. flavogriseus, S. virginiae and S. griseoaurantiacus. It is the first report of endogluconase and xylanolytic activity by S. virginiae isolated from a semi-arid soil.

3-Methyl-2-benzothiazolinone hydrazone and 3-dimethylamino benzoic acid as substrates for the development of polyphenoloxidase and phenoloxidase activity by zymograms.

In the present study, a sequential staining process of polyphenoloxidase and phenoloxidase enzymes was designed by the zymography technique. As a first step, electrophoresis was carried out under native conditions, and later, first staining was carried out with a revealing solution of 3-methyl-2-benzothiazoline hydrazone (MBTH)-3-dimethylamino benzoic acid (DMAB) that allowed the visualization of polyphenoloxidase enzymes, and later and using the same gel, we proceeded to the differential staining of phenoloxidase, adding a solution of H2O2. The technique was standardized using commercial enzymes of laccase (T. versicolor) and horseradish. The technique was used to identify polyphenoloxidases (laccases) and phenoloxidases (lignin peroxidase) of crude extracts obtained from the growth of the basidiomycete Lentinus strigosus on Pinus radiata. The technique showed great sensitivity to detect the different enzymatic activities (1.56 Activity Unit/mL minimum) in the same gel without interference between the enzymes and the solutions used. On the other hand, the efficiency of the technique was compared with the substrates that are commonly used for the detection of this type of activities such as 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and guaiacol, observing greater sensitivity and minimal interference, so that the present method will allow in the same gel, and visualize polyphenoloxidase and phenoloxidase activities simultaneously facilitating expression studies.

Use of barley straw as a support for the production of conidiospores of Trichoderma harzianum.

In this work was to evaluate the conidiospores production of Trichoderma harzianum using barley straw as substrate. Four growth conditions were used; washed and unwashed barley straw and washed and unwashed barley straw supplemented with mineral salts. The highest spore production was observed when washed barley straw supplemented with mineral salts with 1.56 × 1010 conidiospores/gram of dry matter (gdm) at 216 h of cultivation was used. The effect of substrate moisture on spore production was studied, three initial moisture levels of the substrate were tested and it was observed that a humidity of 80 % of the substrate improves the production of conidiospores reaching a concentration of 2.35 × 1010 conidiospores/gdm at 136 h. Finally, conidiospores viability was evaluated for 12 months by keeping them on the conidia and substrate, and viability of 71 % of the conidiospores was observed, so this maintenance method is an excellent means of conserving the conidiospores viability.

Addenbrooke's Cognitive Examination validation in Parkinson's disease.

BACKGROUND: There is a clear need for brief, sensitive and specific cognitive screening instruments in Parkinson's disease (PD). OBJECTIVES: To study Addenbrooke's Cognitive Examination (ACE) validity for cognitive assessment of PD patient's using the Mattis Dementia Rating Scale (MDRS) as reference method. A specific scale for cognitive evaluation in PD, in this instance the Scales for Outcomes of Parkinson's disease-Cognition (SCOPA-COG), as well as a general use scale the Mini-mental state examination (MMSE) were also studied for further correlation. METHODS: Forty-four PD patients were studied, of these 27 were males (61%), with a mean (SD) age of 69.5 (11.8) years, mean (SD) disease duration of 7.6 (6.4) years (range 1-25), mean (SD) total Unified Parkinson's Disease Rating Scale (UPDRS) score 37 (24) points, UPDRS III 16.5 (11.3) points. MDRS, ACE and SCOPA-COG scales were administered in random order. All patients remained in on-state during the study. RESULTS: Addenbrooke's Cognitive Examination correlated with SCOPA-COG (r = 0.93, P < 0.0001), and MDRS (r = 0.91 P < 0.0001) and also with MMSE (r = 0.84, P < 0.001). Area under the receiver-operating curve, taking MDRS as the reference test, was 0.97 [95% confidence interval (CI): 0.92-1.00] for ACE, 0.92 (95% CI: 0.83-1.00) for SCOPA-COG and 0.91 (95% CI: 0.83-1.00) for MMSE. Best cut-off value for ACE was 83 points [Sensitivity (Se) = 92%; Specificity (Sp) = 91%; Kappa concordance (K) = 0.79], 20 points for the SCOPA-COG (Se = 92%; Sp = 87%; K = 0.74) and 26 points for MMSE (Se = 61%; Sp = 100%; K = 0.69). CONCLUSION: Addenbrooke's Cognitive Examination appears to be a valid tool for dementia evaluation in PD, with a cut-off point which should probably be set at 83 points, displaying good correlation with both the scale specifically designed for cognitive deficits in PD namely SCOPA-COG, as well as with less specific tests such as MMSE.

A test method for evaluating the thermal environment of underground coal mine refuge alternatives.

Since 2009, the Mine Safety and Health Administration (MSHA) has required mines to install refuge alternatives (RAs) in underground coal mines. One of the biggest concerns with occupied RAs is the possible severity of the resulting thermal environment. In 30 CFR 7.504, the maximum allowable apparent temperature (AT) for an occupied RA is specified as 35 °C (95 °F). Manufacturers must conduct heat/humidity tests to demonstrate that their RAs meet the 35 °C (95 °F) AT limit. For these tests, heat input devices are used to input the metabolic heat of actual miners. A wide variety of test methods, sensors, and heat input devices could be used when conducting such tests. Since 2012, the National Institute for Occupational Safety and Health (NIOSH) has conducted over thirty 96-hour heat/humidity tests on four different RAs. This paper discusses the test equipment and procedures used during these investigations. This information is useful for RA manufacturers conducting RA heat/humidity tests, for other researchers investigating RA heat/humidity buildup, and for those who need to assess the thermal environment of any confined space where people may be trapped or are seeking refuge.

Hyperhemolytic transfusion reaction attributable to anti-Fy3 in a patient with sickle cell disease.

A case of hyperhemolytic transfusion reaction attributable to anti-Fy3 in a 30-year-oldAfricanAmerican woman with a history of sickle cell disease is reported. The patient was admitted for vaso-occlusive sickle cell crisis and received 4 units of packed RBCs secondary to worsening symptomatic anemia (Hb 5.0 g/dL). On admission, the patient's antibody screen and identification showed anti-V and anti-E, and her antibody history included anti-E,-C,-Jk(b),-N, -V,-S,-Sl(a), and a cold agglutinin with possible anti-I specificity. A DAT performed on her RBCs was negative. RBC units that lacked E, C, Jk(b), N,V, and S were transfused. Posttransfusion Hb was 8.9 g/dL. On day 10 she developed a fever of 103 degrees F, and on day 11 her Hb decreased to 6.4 g/dL. She complained of severe back pain and dark urine. In addition, she became hypertensive, tachycardic, and jaundiced. The DAT indicated the presence of IgG on the patient's RBCs. Anti-Fy3 was identified in the serum and eluate. During the next 24 hours, her Hb decreased to 2.4 g/dL. The LDH level was 1687 U/L, and her reticulocyte count was 2.6%. A delayed hemolytic transfusion reaction with hyperhemolysis secondary to anti-Fy3 was suspected and was successfully treated with IVIG and high-dose prednisone. To the best of our knowledge, this is the first published case of hyperhemolysis in sickle cell disease attributable to anti-Fy3.

Intestinal cell migration damage induced by enteropathogenic Escherichia coli strains.

Epithelial cell migration is an essential response to enteric pathogens such as enteropathogenic Escherichia coli (EPEC). This study aimed to investigate the effects of EPEC infection on intestinal epithelial cell migration in vitro, as well as the involvement of type III secretion system (T3SS) and Rho GTPases. Crypt intestinal epithelial cells (IEC-6) were infected with EPEC strains (E2348/69, ΔescF, and the LDI001 strain isolated from a malnourished Brazilian child) and commensal E. coli HS. Wound migration and cell death assays were performed at different time-points. Transcription and expression of Rho GTPases were evaluated using real-time PCR and western blotting. Overall, EPEC E2348/69 reduced migration and increased apoptosis and necrosis levels compared to EPEC LDI001 and E. coli HS strains. Moreover, EPEC LDI001 impaired cell migration at a higher level than E. coli HS and increased necrosis after 24 hours compared to the control group. The different profiles of virulence genes between the two wild-type EPEC strains, characterized by the absence of espL and nleE genes in the LDI001, might explain the phenotypic results, playing significant roles on cell migration impairment and cell death-related events. Moreover, the type III secretion system is determinant for the inhibition of intestinal epithelial cell migration by EPEC 2348/69, as its deletion prevented the effect. Active Rac1 concentrations were increased in E2348/69 and LDI001-infected cells, while the T3SS-deficient strain did not demonstrate this activation. This study contributes with valuable insight to characterize the mechanisms involved in the impairment of intestinal cell migration induced by EPEC.

Effect of age, malnutrition and renutrition on free secretory component and IgA in secretions.

The development of free secretory component (FSC) was studied in the tears of normal infants, children and adults. The level of FSC in tears was higher in older adults than in children. Free secretory component was also measured in the tears of normal, moderately and severely malnourished Colombian children. Children suffering from kwashiorkor, combined protein-calorie malnutrition or marasmus were studied before and after renutrition. No change was detected in the concentration of FSC in tears of moderately malnourished (Grade I and II) children. There was a significant difference between normal and severely malnourished children which improved with renutrition. The levels of tear IgA were decreased in the moderately malnourished children. These results indicate that reduction in secretory IgA levels in moderate malnutrition may not be explained by a lack of available free secretory component in tears, but that severe malnutrition may impair the S-IgA system by significantly reducing the availability of free secretory component.

Effect of renutrition on humoral and cell-mediated immunity in severely malnourished children.

Forty-three Colombian children suffering from either kwashiorkor (21), combined protein-calorie malnutrition (11), or maramus (11) were hospitalized and provided a high protein, high calorie diet for 4 to 5 wk. Improvement in clinical and nutritional status was accompanied by significant increases in levels of serum immunoglobulins G and M and C3 complement and by significant decreases in serum immunoglobulin A concentrations, especially in infants with kwashiorkor. Skin test reactions to purified protein derivative and candidin improved during renutrition. Lymphocyte blastogenesis after stimulation in vitro with phytohemagglutinin and pokeweed mitogen increased rapidly during hospitalization. After 1 yr posttreatment, cell-mediated immune responses, both in vivo and in vitro, had diminished. These results indicate that some aspects of the immune response are affected to a different degree in kwashiorkor, maramus, and combined malnutrition. Short-term nutritional rehabilitation has a differential effect on the long-term restoration of various aspects of immunity.

Pancreatic and salivary amylase activity in undernourished Colombian children.

Amylase activities were quantitated in secretions of marginally and severely malnourished Colombian children. In young children with a mean age of 21 months, the relative pancreatic and salivary amylase isozyme activities of urine were significantly changed in marginally malnourished children compared to normal children. There was a relative increase in salivary and decrease in pancreatic amylase activity in the undernourished children and total amylase activity was somewhat decreased. Amylase activity in saliva and tears was significantly lower in these malnourished children. Older children who were more severely malnourished had significantly lower amylase activity in their sera and tears. Thus marginal and severe malnutrition affects the production of amylase by the pancreas and salivary glands of young children distinctly. It significantly suppresses amylase activity in tears, saliva, and serum.

Etiology of respiratory tract infections in children in Cali, Colombia.

One hundred eighty children hospitalized for acute respiratory disease were studied in Cali, Colombia. In the majority of patients, pneumonia was the reason for hospitalization and remained the final diagnosis. Fifty-one cases of pneumonia of indeterminate etiology comprised the largest single diagnostic category, followed by 38 cases of pneumonia associated with measles, and 22 cases assocaited with serologic evidence of infection with other viral agents or Mycoplasma pneumoniae. Etiologic diagnosis could be assigned with a reasonable degree of confidence in 116 of the 180 patients (64%). The laboratory procedure found most likely to provide the etiologic diagnosis in this series was paired sera specimens for demonstration of rise in antibody titer against the common viral respiratory pathogens. Those most frequently implicated serologically as etiologic agents in the cases studied were, in order of decreasing frequency, measles, influenza, parainfluenza, and adenoviruses.

Enzootic Brugia infection in a two-year-old Colombian girl.

Mature male and female filarial worms were present in a lymphatic vessel near a retro-auricular lymph node removed from a 2-year old Colombian girl. Although apparently mature microfilariae were present in the uterus of the female, none was detected in blood samples (Knott's) of peripheral blood taken during day or night. The identification of the worms as a species of Brugia is suggested by their location in a dilated lymphatic vessel, morphology of adults, and the distribution pattern of tail nuclei of the microfilariae. This case is similar to another Colombian case reported from the Departamento del Santander del Sur, and suggests that zoonotic filarial infections may not be uncommon in Colombia.

Bilateral tonic-clonic epileptic seizures in non-benign familial neonatal convulsions.

We report an electroclinical pattern considered characteristic of benign familial neonatal convulsions (BFNC) in two neonates without BFNC. Both neonates were products of uncomplicated pregnancies, labor, and deliveries. The cause of the seizures was not established. There was no family history of seizures or epilepsy. Seizures started on the second and third days after birth. Ten seizures were captured by continuous video-EEG telemetry. The electroclinical events began with generalized tonic posturing coinciding with the appearance of diffuse attenuation of the EEG activity. After several seconds, bilateral clonic movements accompanied by bilateral repetitive sharp waves or spikes occurred in the EEG. One patient had normal development; the other became autistic. We conclude that the electroclinical pattern occurring in BFNC can occur in other types of neonatal seizures.

Biological Behavior and Partial Molecular Characterization of Six Chilean Isolates of Plum pox virus.

Plum pox virus (PPV) strain D was first detected in Chile in 1992 infecting Prunus trees including peaches, nectarines, apricots, and plums. Since then, quarantine efforts have included periodic surveys in the central zone of the country, the main region for stone fruit production. This work describes the characterization of six PPV isolates from this area of Chile, using biological and molecular approaches. PPV isolates were introduced into Prunus tomentosa and Nicotiana benthamiana hosts by grafting and mechanical inoculation, respectively. Symptoms were evaluated by following the appearance of circular necrotic spots and mosaic in leaves of P. tomentosa and mosaic and some leaf deformation in N. benthamiana. Molecular analysis was carried out using reverse transcription-polymerase chain reaction, allowing the cloning and sequencing of 1.34-kb fragments corresponding to the 3' region of the replicase gene, the complete coat protein (CP) gene, and the 3' nontranslated region of the PPV genome. Evolutionary distance analysis of these nucleotide sequences and their deduced coat protein amino acid sequences grouped the six Chilean isolates among strain D isolates, with closest genetic distances to those of Central Germany and Poland. Representative sources of these isolates suggest that strain D could be the only type of PPV currently present in Chile.

[Hypocaloric peripheral parenteral nutrition]. / Nutrición parenteral periférica hipocalórica.

We carried out a prospective study of a group of 84 patients entering the Digestive Surgery Service for surgery, dividing them at random into two groups, one fed with peripheral parenteral nutrition (PPN) and the other with conventional serum therapy during the period of post-operative fasting. The aim of this study was to see whether PNN can provide significant benefits from a nutritional stand-point in comparison with the traditional serum treatment used until now. We appraised the nutritional state immediately prior to the operation, at the time of hospital admission, and compared it with that one week following the operation. We found improvements in the PNN group. The deterioration following surgery occurred in the two groups in significant form, although less in patients treated with PNN, in whom there was no variation in the nutritional state on admission and one week following surgery, while such a variation did occur in those undergoing serum therapy. PNN is a good option where the digestive pipe cannot be used as nutrient input, for a period of time which is not prolonged, but different solutions must continue to be examined which provide a greater nutritional substrate that that used by us.

[Incidence of malnutrition in surgical departments of the Reina Sofía Hospital of Córdoba]. / Incidencia de la desnutrición en los servicios quirúrgicos del Hospital Reina Sofía de Córdoba.

We conducted a prospective study of the incidence of malnutrition in the surgical departments of the Reina Sofia University Hospital, on a total of 329 patients at the time of admittance (154 women and 175 men). As was denounced by Bristian more than 20 years ago, more than 50% of the patients presented some form of malnutrition. There are a series of factors, detected by the Chi2 test, which are as divers as sex, marital status, level of education, profession, smoker or chronic medication user, which are of influence on the nutritional status of the patients. The type of malnutrition is conditioned by the marital status, the job situation, the nature of the pathology, or the use of tobacco. Malnutrition still exists in developed countries, induced by many factors, and it is often found in the hospitals, which is even worse.

[Nutritional status of patients undergoing digestive surgery]. / Estado nutricional de los pacientes de cirugía digestiva.

A retrospective study of 102 patients, analysing the incidence of malnutrition, was done in the Digestive Surgery Department of the Queen Sophia University Hospital, Cordoba. Patients were subjected to a protocol of nutritional evaluation to find out their nutritional condition and type and degree of malnutrition. The high incidence of malnutrition sufferers (> 80%) at the time of hospital admission was confirmed, being more noticeable in neoplastic patients.

[Complications of TPN in gastrointestinal surgery patients]. / Complicaciones de la NPT en pacientes sometidos a cirugía gastrointestinal.

Retrospective study of a group of 77 patients subjected to gastrointestinal surgery, who received total parenteral nutrition (TPN) during the postoperative period, During the evolution of the nutrition, 76 cases with technical complications arose, one case of sepsis, 10 cases of metabolic complications and one patient with haematological complications (acute medular failure). Although a higher percentage of metabolic and haematological alterations not requiring medical or surgical treatment for correction was observed, these were indicative of the tendencies of the metabolic and haematological states of the patients. An analysis was also made of the variance, observing a direct relationship between the components of TPN and the variations of the metabolic and haematological values studied, However, these values did not correlate with the number of days during which the patients was on nutrition, and thus the technique and guidelines for administration are the most important factors which determine the presence or absence of complications during TPN.

Intestinal cell migration damage induced by enteropathogenic Escherichia coli strains

Epithelial cell migration is an essential response to enteric pathogens such as enteropathogenic Escherichia coli (EPEC). This study aimed to investigate the effects of EPEC infection on intestinal epithelial cell migration in vitro, as well as the involvement of type III secretion system (T3SS) and Rho GTPases. Crypt intestinal epithelial cells (IEC-6) were infected with EPEC strains (E2348/69, ΔescF, and the LDI001 strain isolated from a malnourished Brazilian child) and commensal E. coli HS. Wound migration and cell death assays were performed at different time-points. Transcription and expression of Rho GTPases were evaluated using real-time PCR and western blotting. Overall, EPEC E2348/69 reduced migration and increased apoptosis and necrosis levels compared to EPEC LDI001 and E. coli HS strains. Moreover, EPEC LDI001 impaired cell migration at a higher level than E. coli HS and increased necrosis after 24 hours compared to the control group. The different profiles of virulence genes between the two wild-type EPEC strains, characterized by the absence of espL and nleE genes in the LDI001, might explain the phenotypic results, playing significant roles on cell migration impairment and cell death-related events. Moreover, the type III secretion system is determinant for the inhibition of intestinal epithelial cell migration by EPEC 2348/69, as its deletion prevented the effect. Active Rac1 concentrations were increased in E2348/69 and LDI001-infected cells, while the T3SS-deficient strain did not demonstrate this activation. This study contributes with valuable insight to characterize the mechanisms involved in the impairment of intestinal cell migration induced by EPEC.