Immunological Cross-Reactivity and Preclinical Assessment of a Colombian Anticoral Antivenom against the Venoms of Three Micrurus Species.

Snakebite accident treatment requires the administration of antivenoms that provide efficacy and effectiveness against several snake venoms of the same genus or family. The low number of immunogenic components in venom mixtures that allow the production of antivenoms consequently gives them partial neutralization and a suboptimal pharmacological response. This study evaluates the immunorecognition and neutralizing efficacy of the polyvalent anticoral antivenom from the Instituto Nacional de Salud (INS) of Colombia against the heterologous endemic venoms of Micrurus medemi, and M. sangilensis, and M. helleri by assessing immunoreactivity through affinity chromatography, ELISA, Western blot, and neutralization capability. Immunorecognition towards the venoms of M. medemi and M. sangilensis showed values of 62% and 68% of the protein composition according to the immunoaffinity matrix, respectively. The analysis by Western blot depicted the highest recognition patterns for M. medemi, followed by M. sangilensis, and finally by M. helleri. These findings suggest that the venom compositions are closely related and exhibit similar recognition by the antivenom. According to enzyme immunoassays, M. helleri requires a higher amount of antivenom to achieve recognition than the others. Besides reinforcing the evaluation of INS antivenom capability, this work recommends the use of M. helleri in the production of Colombian antisera.

Inhibition of the NMDA Currents by Probenecid in Amygdaloid Kindling Epilepsy Model.

Epilepsy is characterized by a sustained depolarization and repeated discharge of neurons, attributed to overstimulation of N-methyl-D-aspartate receptors (NMDAr). Herein, we propose that probenecid (PROB), an inhibitor of the activity of some ATP binding-cassette transporters (ABC-transporters) can modify NMDAr activity and expression in amygdaloid kindled model. Some studies have suggested that NMDAr expression could be regulated by inhibiting the activity of P-glycoprotein (MDR1) and drug resistance protein-1 (MRP1). Besides, PROB was found to interact with other proteins with proven activity in the kindling model, such as TRPV2 channels, OAT1, and Panx1. Administering PROB at two doses (100 and 300 mg/kg/d) for 5 d decreased after-discharge duration and Racine behavioral scores. It also reduced the expression of NR2B and the activity of total NOS and the expression of nNOS with respect to the kindling group. In a second protocol, voltage-clamp measurements of NMDA-evoked currents were performed in CA1 hippocampal cells dissociated from control and kindled rats. PROB produced a dose-dependent reduction in NMDA-evoked currents. In neurons from kindled rats, a residual NMDA-evoked current was registered with respect to control animals, while a reduction in NMDA-evoked currents was observed in the presence of 20 mM PROB. Finally, we evaluated the expression of MRP1 and MDR1 in order to establish a relationship between the reduction of kindling parameters, the inhibition of NMDA-type currents, and the expression of these transporters. Based on our results, we conclude that at the concentrations used, PROB inhibits currents evoked by NMDA in dissociated neurons of control and kindled rats. In the kindling model, at the tested doses, PROB decreases the after-discharge duration and Racine behavioral score in the kindling model. We propose a mechanism that could be dependent on the expression of ABC-type transporters.

Melatonin in Combination with Albendazole or Albendazole Sulfoxide Produces a Synergistic Cytotoxicity against Malignant Glioma Cells through Autophagy and Apoptosis.

Glioblastoma is the most aggressive and lethal brain tumor in adults, presenting diffuse brain infiltration, necrosis, and drug resistance. Although new drugs have been approved for recurrent patients, the median survival rate is two years; therefore, new alternatives to treat these patients are required. Previous studies have reported the anticancer activity of albendazole, its active metabolite albendazole sulfoxide, and melatonin; therefore, the present study was performed to evaluate if the combination of melatonin with albendazole or with albendazole sulfoxide induces an additive or synergistic cytotoxic effect on C6 and RG2 rat glioma cells, as well as on U87 human glioblastoma cells. Drug interaction was determined by the Chou-Talalay method. We evaluated the mechanism of cell death by flow cytometry, immunofluorescence, and crystal violet staining. The cytotoxicity of the combinations was mainly synergistic. The combined treatments induced significantly more apoptotic and autophagic cell death on the glioma cell lines. Additionally, albendazole and albendazole sulfoxide inhibited proliferation independently of melatonin. Our data justify continuing with the evaluation of this proposal since the combinations could be a potential strategy to aid in the treatment of glioblastoma.

Chronic Consumption of Fructose Induces Behavioral Alterations by Increasing Orexin and Dopamine Levels in the Rat Brain.

It has been widely described that chronic intake of fructose causes metabolic alterations which can be associated with brain function impairment. In this study, we evaluated the effects of fructose intake on the sleep⁻wake cycle, locomotion, and neurochemical parameters in Wistar rats. The experimental group was fed with 10% fructose in drinking water for five weeks. After treatment, metabolic indicators were quantified in blood. Electroencephalographic recordings were used to evaluate the sleep architecture and the spectral power of frequency bands. Likewise, the locomotor activity and the concentrations of orexin A and monoamines were estimated. Our results show that fructose diet significantly increased the blood levels of glucose, cholesterol, and triglycerides. Fructose modified the sleep⁻wake cycle of rats, increasing the waking duration and conversely decreasing the non-rapid eye movement sleep. Furthermore, these effects were accompanied by increases of the spectral power at different frequency bands. Chronic consumption of fructose caused a slight increase in the locomotor activity as well as an increase of orexin A and dopamine levels in the hypothalamus and brainstem. Specifically, immunoreactivity for orexin A was increased in the ventral tegmental area after the intake of fructose. Our study suggests that fructose induces metabolic changes and stimulates the activity of orexinergic and dopaminergic neurons, which may be responsible for alterations of the sleep⁻wake cycle.

[Grooming, Cyberbullying and Sexting in Chile according of sex and school management or administrative dependency]. / Grooming, Ciberbullying y Sexting en estudiantes en Chile según sexo y tipo de administración escolar.

INTRODUCTION: Children, teenagers and young men are increasingly experiencing their well-being related to the internet and the new digital technologies. The objective of this study is to describe the presence of Cyberbullying, Sexting and Grooming in students in Chile according to gender and type of school management or administrative dependency. SUBJECTS AND METHOD: Exploratory and descriptive study. The sample design was non-probabilistic by quotas in 60 transactional establish ments. The sample was weighted considering the age range and gender according to national data. The Digital Literacy Questionnaire "Divergente-SerDigital" (2010) was applied to a sample of 12,926 students, aged 5 to 18 years. 4,790 men and 8,136 women. Average age 13.17 years. Frequencies were analyzed and the Chi-squared contrast statistic was used to determine statistically significant differences. RESULTS: The item Total Grooming (cheating) is presented as the main risk, 12.6% in municipal dependent schools (MDS), 8.2% in subsidized private schools (SPS), and 8.4% in private schools (PS). When considering gender, Grooming is observed mainly in Men, 20.4% in MDS, 19.9% in SPS and, 16.9% in PS. It is noteworthy that Women perform less Cyberbullying (active) according to school administration with 4.2% in MDS, 2.4% in SPS and, 2.6% in PS, with statistically significant differences (p < 0.05) in relation to Men. It also highlights the indicator Sexting (send) in Men, higher in PS with 10.6%. CONCLUSION: Grooming, Cyberbullying and Sexting risks are presented in the three types of administration with specific characteristics. These data can be a guide to work in promotion and prevention as well as in the schematization of cases according to type of school administration.

Grooming, ciberbullying y sexting en estudiantes en Chile según sexo y tipo de administración escolar / Grooming, cyberbullying and sexting in Chile according of sex and school management or administrative dependency

INTRODUCCIÓN: Niños y adolescentes experimentan su bienestar cada día más relacionado con internet y las nuevas tecnologías digitales. El objetivo del manuscrito es describir la presencia de Ciberbullying (acoso o agresión entre menores o pares en internet), Sexting (difundir intimidad sexual) y Groo ming (engaño online a menores de edad por parte de adultos) en los estudiantes en Chile según sexo y tipo de administración escolar. SUJETOS Y MÉTODO: Estudio de carácter exploratorio y descriptivo. El diseño muestral fue no probabilístico por cuotas en 60 establecimientos de carácter transaccional. La muestra se ponderó considerando rango de edad y sexo según datos nacionales. Se aplicó el Cuestionario de Alfabetización Digital "Divergente-SerDigital" (2010) a una muestra de 12.926 estudiantes, rango de edad: 5 a 18 años. 4.790 hombres y 8.136 mujeres. Edad promedio 13,17 años. Se analizaron frecuencias y se utilizó el estadístico de contraste Chi cuadrado para determinar diferencias estadísticamente significativas. RESULTADOS: El ítem Grooming total (engaño) se presenta como el principal riesgo, 12,6% en Colegios Municipales (CM), 8,2% en Colegios Particulares Subvencionados (CPS) y 8,4% Colegios Particulares Privados (CPP). Al considerar el sexo se observa Grooming principal mente en Hombres, 20,4% en CM, 19,9% CPS y 16,9% CPP. Destaca que las Mujeres realizan menos Ciberbullying (activo) según administración escolar con 4,2% en CM, 2,4% CPS y 2,6% CPP, con diferencias estadísticamente significativa (p < 0,05) en relación a los Hombres. Además destaca el indicador Sexting (enviar) en Hombres, más alto en los CPP con 10,6%. CONCLUSIÓN: Los riesgos Grooming, Ciberbullying y Sexting se presentan en los tres tipos de administración con características específicas. Estos datos pueden ser guía del trabajo en promoción y prevención como en la tematización de casos según tipo de administración escolar.

INTRODUCTION: Children, teenagers and young men are increasingly experiencing their well-being related to the internet and the new digital technologies. The objective of this study is to describe the presence of Cyberbullying, Sexting and Grooming in students in Chile according to gender and type of school management or administrative dependency. SUBJECTS AND METHOD: Exploratory and descriptive study. The sample design was non-probabilistic by quotas in 60 transactional establish ments. The sample was weighted considering the age range and gender according to national data. The Digital Literacy Questionnaire "Divergente-SerDigital" (2010) was applied to a sample of 12,926 students, aged 5 to 18 years. 4,790 men and 8,136 women. Average age 13.17 years. Frequencies were analyzed and the Chi-squared contrast statistic was used to determine statistically significant differences. RESULTS: The item Total Grooming (cheating) is presented as the main risk, 12.6% in municipal dependent schools (MDS), 8.2% in subsidized private schools (SPS), and 8.4% in private schools (PS). When considering gender, Grooming is observed mainly in Men, 20.4% in MDS, 19.9% in SPS and, 16.9% in PS. It is noteworthy that Women perform less Cyberbullying (active) according to school administration with 4.2% in MDS, 2.4% in SPS and, 2.6% in PS, with statistically significant differences (p < 0.05) in relation to Men. It also highlights the indicator Sexting (send) in Men, higher in PS with 10.6%. CONCLUSION: Grooming, Cyberbullying and Sexting risks are presented in the three types of administration with specific characteristics. These data can be a guide to work in promotion and prevention as well as in the schematization of cases according to type of school administration.

Participation of the dentate-rubral pathway in the kindling model of epilepsy.

Lesions of the cerebellar dentate nucleus (DN) reduce the after-discharge duration induced by repetitive kindling stimulation and decrease seizures to a lower rank according to Racine's scale. The DN sends cholinergic and glutamatergic fibers to the red nucleus (RN), which is composed of glutamatergic and GABAergic cells. To test the participation of these neurotransmitters in seizures, we compared the levels of glutamate and gamma-aminobutyric acid (GABA) at the RN in a control condition, a kindled stage, and a kindled stage followed by DN lesions. We found that the kindled stage was associated with significant reductions in glutamate and GABA in the RN and that the lesions of the DN in kindled rats reversed the severity of seizures and restored the GABA levels. GAD65 , a GABA-synthesizing enzyme, was increased in kindled rats and decreased after DN lesions. GAD65 commonly appears localized at nerve terminals and synapses, and it is only activated when GABA neurotransmission occurs. Thus, it is possible that the increased expression of GAD65 found in kindled rats could be due to an exacerbated demand for GABA due to kindled seizures. It is known that GABA maintains the inhibitory tone that counterbalances neuronal excitation. The decreased expression of GAD65 found after the DN lesions indicated that the GABA-synthesizing enzyme was no longer required once it eliminated the excitatory glutamate input to the RN. We thus conclude that DN lesions and their consequent biochemical changes are capable of decreasing the generalized seizures induced by kindling stimulation. © 2016 Wiley Periodicals, Inc.

Exposure to ozone induces a systemic inflammatory response: possible source of the neurological alterations induced by this gas.

The World Health Organization identified urban outdoor air pollution as the eighth highest mortality risk factor in high-income countries. Exposure to ambient pollutants such as ozone (O3) increases the number of hospital admissions. O3 is a highly reactive gas that reacts with cells lining the airways, producing the formation of reactive oxygen species and inflammation. Beyond the respiratory system, O3 exposure also produces fatigue, lethargy, headaches, and significant decrease in rapid-eye-movement sleep related to an increase in slow-wave sleep. Interestingly, these sleep changes can be significantly mitigated by treatment with indomethacin, which suggests that an inflammatory mechanism may be responsible for these neurological symptoms. To characterize the inflammatory mechanisms by which O3 affects tissues outside the pulmonary system, we evaluated inflammatory factors in both lung and brain. Rats exposed to 1 part per million O3 for 1, 3 or 6 h, as well as rats exposed daily for 1 or 3 h over five consecutive days, showed increases in TNF-α and IL-6 levels within the lungs as well as increases in TNF-α, IL-6, NF-κB p50 and GFAP levels in the cerebral cortex. These results support the hypothesis that the neuroinflammatory response may be responsible for the central nervous system effects of O3 exposure.

The effects of ozone exposure and associated injury mechanisms on the central nervous system.

Ozone (O3) is a component of photochemical smog, which is a major air pollutant and demonstrates properties that are harmful to health because of the toxic properties that are inherent to its powerful oxidizing capabilities. Environmental O3 exposure is associated with many symptoms related to respiratory disorders, which include loss of lung function, exacerbation of asthma, airway damage, and lung inflammation. The effects of O3 are not restricted to the respiratory system or function - adverse effects within the central nervous system (CNS) such as decreased cognitive response, decrease in motor activity, headaches, disturbances in the sleep-wake cycle, neuronal dysfunctions, cell degeneration, and neurochemical alterations have also been described; furthermore, it has also been proposed that O3 could have epigenetic effects. O3 exposure induces the reactive chemical species in the lungs, but the short half-life of these chemical species has led some authors to attribute the injurious mechanisms observed within the lungs to inflammatory processes. However, the damage to the CNS induced by O3 exposure is not well understood. In this review, the basic mechanisms of inflammation and activation of the immune system by O3 exposure are described and the potential mechanisms of damage, which include neuroinflammation and oxidative stress, and the signs and symptoms of disturbances within the CNS caused by environmental O3 exposure are discussed.

Parallel microwave-assisted synthesis of ionic liquids and screening for denitrogenation of straight-run diesel feed by liquid-liquid extraction.

Fifty-six ionic liquids were efficiently synthesized in parallel format under one-pot, solvent-free microwave-assisted synthesis. These compounds were evaluated as extracting agents of nitrogen-containing compounds from a real Diesel feed before being submitted to the hydrodesulfurization process to obtain ultralow sulfur Diesel. Our results showed that halogenated ionic liquids are an excellent alternative due to these ionic liquids are relatively inexpensive, presenting a high selectivity for the extraction of nitrogen-containing compounds and can be regenerated and recycled.

Insulin analogs versus human insulin in the treatment of patients with diabetic ketoacidosis: a randomized controlled trial.

OBJECTIVE: To compare the safety and efficacy of insulin analogs and human insulins both during acute intravenous treatment and during the transition to subcutaneous insulin in patients with diabetic ketoacidosis (DKA). RESEARCH DESIGN AND METHODS: In a controlled multicenter and open-label trial, we randomly assigned patients with DKA to receive intravenous treatment with regular or glulisine insulin until resolution of DKA. After resolution of ketoacidosis, patients treated with intravenous regular insulin were transitioned to subcutaneous NPH and regular insulin twice daily (n = 34). Patients treated with intravenous glulisine insulin were transitioned to subcutaneous glargine once daily and glulisine before meals (n = 34). RESULTS: There were no differences in the mean duration of treatment or in the amount of insulin infusion until resolution of DKA between intravenous treatment with regular and glulisine insulin. After transition to subcutaneous insulin, there were no differences in mean daily blood glucose levels, but patients treated with NPH and regular insulin had a higher rate of hypoglycemia (blood glucose <70 mg/dl). Fourteen patients (41%) treated with NPH and regular insulin had 26 episodes of hypoglycemia and 5 patients (15%) in the glargine and glulisine group had 8 episodes of hypoglycemia (P = 0.03). CONCLUSIONS: Regular and glulisine insulin are equally effective during the acute treatment of DKA. A transition to subcutaneous glargine and glulisine after resolution of DKA resulted in similar glycemic control but in a lower rate of hypoglycemia than with NPH and regular insulin. Thus, a basal bolus regimen with glargine and glulisine is safer and should be preferred over NPH and regular insulin after the resolution of DKA.

Comparison of inpatient insulin regimens with detemir plus aspart versus neutral protamine hagedorn plus regular in medical patients with type 2 diabetes.

BACKGROUND: Studies comparing the use of basal bolus with insulin analogs vs. split-mixed regimens with human insulins in hospitalized patients with type 2 diabetes are lacking. RESEARCH DESIGN AND METHODS: In a controlled multicenter trial, we randomized 130 nonsurgical patients with blood glucose (BG) between 140 and 400 mg/dl to receive detemir once daily and aspart before meals (n = 67) or neutral protamine Hagedorn (NPH) and regular insulin twice daily (n = 63). Insulin dose was started at 0.4 U/kg.d for BG between 140 and 200 mg/dl or 0.5 U/kg.d for BG 201-400 mg/dl. Major study outcomes included differences in mean daily BG levels and frequency of hypoglycemic events between treatment groups. RESULTS: Glycemic control improved similarly in both groups from a mean daily BG of 228 +/- 54 and 223 +/- 58 mg/dl (P = 0.61) to a mean daily BG level after the first day of 160 +/- 38 and 158 +/- 51 mg/dl in the detemir/aspart and NPH/regular insulin groups, respectively (P = 0.80). A BG target below 140 mg/dl before meals was achieved in 45% of patients in the detemir/aspart group and 48% in the NPH/regular group (P = 0.86). During treatment, 22 patients (32.8%) in the detemir/aspart group and 16 patients (25.4%) in the NPH/regular group had at least one episode of hypoglycemia (BG < 60 mg/dl) during the hospital stay (P = 0.34). CONCLUSIONS: Treatment with basal/bolus regimen with detemir once daily and aspart before meals results in equivalent glycemic control and no differences in the frequency of hypoglycemia compared to a split-mixed regimen of NPH and regular insulin in patients with type 2 diabetes.

Prevalence and clinical outcome of inpatient hyperglycemia in a community pediatric hospital.

BACKGROUND: Inpatient hyperglycemia in adult patients with and without a history of diabetes is a predictor of poor clinical outcome. No previous studies, however, have examined the association of hyperglycemia and clinical outcome in children admitted to a community pediatric hospital. METHODS: The study was a retrospective observational cohort of pediatric patients admitted to a community children's hospital from January 2004 to August 2004. Medical records of 903 consecutive children admitted to critical and non-critical care areas were reviewed. Of them, 342 patients (38%) had no blood glucose measurements during their hospital stay. In the remaining patients, we determined the prevalence of hyperglycemia and examined the association of hyperglycemia with clinical outcome. RESULTS: A total of 406 patients (75%) had an admission blood glucose < or =120 mg/dL (mean +/- SEM 98 +/- 1 mg/dL), 103 children (19%) had an admission blood glucose level of 121-179 mg/dL (mean 143 +/- 2 mg/dL), and 32 patients (5.9%) had a blood glucose level > or =180 mg/dL (mean 260 +/- 18 mg/dL). Seventeen patients (13%) had a known history of diabetes prior to admission. Children with hyperglycemia were more likely to be admitted to the ICU (P < .001) and had a longer length of ICU stay (P < .001), but admission hyperglycemia was not associated with longer hospital stay or higher hospital mortality. CONCLUSIONS: Hyperglycemia is present in one-fourth of children admitted to the hospital, most of them without a history of diabetes prior to admission. Hyperglycemia was associated with a greater need for ICU care and longer ICU stay but not with increased in-hospital mortality.

Randomized study of basal-bolus insulin therapy in the inpatient management of patients with type 2 diabetes (RABBIT 2 trial).

OBJECTIVE: We sought to study the optimal management of hyperglycemia in non-intensive care unit patients with type 2 diabetes, as few studies thus far have focused on the subject. RESEARCH DESIGN AND METHODS: We conducted a prospective, multicenter, randomized trial to compare the efficacy and safety of a basal-bolus insulin regimen with that of sliding-scale regular insulin (SSI) in patients with type 2 diabetes. A total of 130 insulin-naive patients were randomized to receive glargine and glulisine (n = 65) or a standard SSI protocol (n = 65). Glargine was given once daily and glulisine before meals at a starting dose of 0.4 units x kg(-1) x day(-1) for blood glucose 140-200 mg/dl or 0.5 units x kg(-1) x day(-1) for blood glucose 201-400 mg/dl. SSI was given four times per day for blood glucose >140 mg/dl. RESULTS: The mean admission blood glucose was 229 +/- 6 mg/dl and A1C 8.8 +/- 2%. A blood glucose target of <140 mg/dl was achieved in 66% of patients in the glargine and glulisine group and in 38% of those in the SSI group. The mean daily blood glucose between groups ranged from 23 to 58 mg/dl, with an overall blood glucose difference of 27 mg/dl (P < 0.01). Despite increasing insulin doses, 14% of patients treated with SSI remained with blood glucose >240 mg/dl. There were no differences in the rate of hypoglycemia or length of hospital stay. CONCLUSIONS: Treatment with insulin glargine and glulisine resulted in significant improvement in glycemic control compared with that achieved with the use of SSI alone. Our study indicates that a basal-bolus insulin regimen is preferred over SSI in the management of non-critically ill, hospitalized patients with type 2 diabetes.

The 19-kDa antigen of Mycobacterium tuberculosis is a major adhesin that binds the mannose receptor of THP-1 monocytic cells and promotes phagocytosis of mycobacteria.

Identification of mycobacterial adhesins is needed to understand better the pathogenesis of tuberculosis and to develop new strategies to fight this infection. In this work, THP-1 monocytic cells were incubated with Mycobacterium tuberculosis culture filtrate proteins labelled with biotin and a dominant 19-kDa adhesin was found. This adhesin was characterized as the glycosylated and acylated 19-kDa antigen (Rv 3763). These findings were confirmed in assays with culture filtrate proteins and cell-wall fractions from a recombinant Mycobacterium smegmatis strain that overexpresses the 19-kDa antigen. Further, fluorescent microspheres coated with recombinant culture filtrate proteins adhere to cells in higher numbers than microspheres coated with native M. smegmatis proteins. The binding of the 19-kDa antigen to cells was inhibited with mannose receptor competitor sugars, Ca(2+) chelators and with a monoclonal antibody to the human mannose receptor. Phagocytosis assays showed high-level binding of bacilli to THP-1 cells that was inhibited with alpha-methyl-mannoside, mannan, EDTA and mAbs to the mannose receptor and to the 19-kDa M. tuberculosis antigen. Immunoprecipitation, cell-surface ELISA and immunostaining confirmed the expression of the mannose receptor by THP-1 cells. In conclusion, here we show that the macrophage mannose receptor, considered a pathogen pattern recognition receptor, may interact with mannose residues of mycobacterial glycoproteins that could promote the phagocytosis of mycobacteria.

Fasceitis necrotizante después del nacimiento por cesárea / Necrotizing fasciitis later of the birth by cesarean

Informamos de un caso de fasceitis necrotizante posterior a una cesárea.