Alterations of the erythrocyte membrane by in vitro action of Trichinella spiralis muscle larvae.

The complex life cycle of Trichinella spiralis includes the migration of newborn larvae through the bloodstream to their encystment in muscle. The parasite establishes an intimate contact with the erythrocytes of the host both during the migration of the newborn larvae and when encysting, as this parasite causes intense vascularization in the muscle cell. The goal of this work was to study the effects of various concentrations of T. spiralis muscle larvae (ML) on erythrocyte membranes. The treatment was performed by incubating human erythrocytes with equal volume of different concentrations of ML for 30 minutes, with controlled agitation (37°C). The control erythrocytes (with no contact with the larvae) were incubated in the same way with an equal volume of physiological solution. To evaluate the alterations to the erythrocytes by the action of the larvae and in the respective controls, an Erythrocyte Rheometer and a Digital Image Analysis technique were used. The results indicated that when the larval concentration was higher, the aggregation and erythrocyte membrane alterations were also higher. Also, the erythrocyte deformability index and the erythrocyte elasticity increased. The values of isolated cell coefficient varied from 0.51 in the treatment with 100 larvae/ml to 0.91 in the incubation with 1000 larvae/ml. This experiment shows that T. spiralis muscle larvae affect significantly the red blood cell aggregation and the erythrocyte viscoelastic properties.

Human presence impacts fungal diversity of inflated lunar/Mars analog habitat.

BACKGROUND: An inflatable lunar/Mars analog habitat (ILMAH), simulated closed system isolated by HEPA filtration, mimics International Space Station (ISS) conditions and future human habitation on other planets except for the exchange of air between outdoor and indoor environments. The ILMAH was primarily commissioned to measure physiological, psychological, and immunological characteristics of human inhabiting in isolation, but it was also available for other studies such as examining its microbiological aspects. Characterizing and understanding possible changes and succession of fungal species is of high importance since fungi are not only hazardous to inhabitants but also deteriorate the habitats. Observing the mycobiome changes in the presence of human will enable developing appropriate countermeasures with reference to crew health in a future closed habitat. RESULTS: Succession of fungi was characterized utilizing both traditional and state-of-the-art molecular techniques during the 30-day human occupation of the ILMAH. Surface samples were collected at various time points and locations to observe both the total and viable fungal populations of common environmental and opportunistic pathogenic species. To estimate the cultivable fungal population, potato dextrose agar plate counts method was utilized. The internal transcribed spacer region-based iTag Illumina sequencing was employed to measure the community structure and fluctuation of the mycobiome over time in various locations. Treatment of samples with propidium monoazide (PMA; a DNA intercalating dye for selective detection of viable microbial populations) had a significant effect on the microbial diversity compared to non-PMA-treated samples. Statistical analysis confirmed that viable fungal community structure changed (increase in diversity and decrease in fungal burden) over the occupation time. Samples collected at day 20 showed distinct fungal profiles from samples collected at any other time point (before or after). Viable fungal families like Davidiellaceae, Teratosphaeriaceae, Pleosporales, and Pleosporaceae were shown to increase during the occupation time. CONCLUSIONS: The results of this study revealed that the overall fungal diversity in the closed habitat changed during human presence; therefore, it is crucial to properly maintain a closed habitat to preserve it from deteriorating and keep it safe for its inhabitants. Differences in community profiles were observed when statistically treated, especially of the mycobiome of samples collected at day 20. On a genus level Epiccocum, Alternaria, Pleosporales, Davidiella, and Cryptococcus showed increased abundance over the occupation time.

Isolated, well-defined organovanadium(iii) on silica: single-site catalyst for hydrogenation of alkenes and alkynes.

A well-defined, isolated, single-site organovanadium(iii) catalyst on SiO2 [(SiO2)V(Mes)(THF)] was synthesized via surface organometallic chemistry, and fully characterized using a combination of analytical and spectroscopic techniques (EA, ICP, 1H NMR, TGA-MS, EPR, XPS, DR-UV/Vis, UV-Raman, DRIFTS, XAS). The catalyst exhibits unprecedented reactivity in liquid- and gas-phase alkene/alkyne hydrogenation. Kinetic poisoning experiments revealed that 100% of the V sites are active for hydrogenation.

Modulation of IL-12 and IFNγ by probiotic supplementation promotes protection against Toxocara canis infection in mice.

In this study, supplementation with the probiotic Saccharomyces boulardii promoted a reduction in intensity of infection by Toxocara canis and modulates cytokines mRNA expression in experimentally infected mice. IL-12 gene transcription had 40-fold increase in S. boulardii supplemented uninfected mice and sevenfold increase in supplemented infected mice comparing with not supplemented group. Regarding IFNγ, similar results were observed, since probiotic supplementation induced approximately 43-fold increase, but only in uninfected mice (P < 0·05). T. canis infection upregulated IL-10 expression while S. boulardii downregulated it and no change was observed for IL-4. Thus, based in these findings; we suggest that one possible mechanism responsible for S. boulardii protection effect against T. canis infection is by the modulation of cytokines expression, especially IL-12.

First record of a spawning aggregation for the tropical eastern Pacific endemic grouper Mycteroperca olfax in the Galapagos Marine Reserve.

This study provides direct and indirect evidence of temporally and spatially consistent spawning aggregations for the grouper Mycteroperca olfax. Recently reported declines in population numbers, probably related to the direct targeting of aggregations by artisanal fishermen, highlight the urgent need for species-specific management actions in the Galapagos Marine Reserve, such as minimum and maximum landing sizes, and the importance of protecting key aggregation sites with the declaration of no-take areas and the establishment of total fishing bans during the reproductive season.

Cryopreservation of immature equine oocytes, comparing a solid surface vitrification process with open pulled straws and the use of a synthetic ice blocker.

The objective was to evaluate the effect of three cryopreservation methods on the in vitro maturation (IVM) and membrane integrity (MIn) of immature equine oocytes. An open pulled straw (OPS) method, a novel solid surface vitrification (SSV) process, and the addition of a synthetic ice blocker were evaluated. Compared with the control group (N=269), the OPS (N=159) and the SSV (N=202) cryopreservation methods decreased both IVM (50.9 vs. 13.3 and 9.4%, respectively; P<0.001) and MIn (76.6 vs. 31.1 and 33.7%; P<0.001) of immature equine oocytes. However, inclusion of 0.1% ice blocker in the OPS vitrification process increased the rates of both IVM (30.5%; P<0.01) and MIn (45.8%; P<0.05) of the oocytes (N=59). Including 0.1% ice blocker in the SSV process improved the IVM rate (20.9%; P<0.05), whereas MIn remained compromised in this group (N=67). However, increasing the concentration of the ice blocker (to 1.0%) in the cryopreservation methods did not significantly improve rates of IVM. In conclusion, the addition of a synthetic ice blocker (0.1%) to both cryopreservation processes significantly increased rates of both IVM and MIn of immature equine oocytes cryopreserved by OPS.

NanoSMGT: transfection of exogenous DNA on sex-sorted bovine sperm using nanopolymer.

The objective was to introduce exogenous DNA into commercially sex-sorted bovine sperm using nanopolymer for transfection. In the first experiment, the optimal concentration and ratio of linear-to-circular plasmid was determined for NanoSMGT in unsorted sperm. A second experiment was conducted to transfect exogenous DNA into sex-sorted sperm. Exogenous DNA uptake occurred in a dose-dependent manner (P < 0.05). The optimal amount of DNA was 10 µg/10(6) cells. The ratios of linear-to-circular plasmid do not influence the uptake by unsorted sperm cells and none of the tested treatments affected sperm motility and viability. Commercially sex-sorted bovine sperm were able to uptake exogenous DNA using nanopolymer; however, both X- and Y-sorted sperm had decreased DNA uptake in comparison to unsorted sperm (P < 0.05). Neither sperm motility nor viability were affected by nanotransfection. In conclusion, nanopolymer efficiently introduced exogenous DNA into commercially sex-sorted bovine sperm; we inferred that these sperm could be used for production of embryos of the desired sex, a technique named NanoSMGT.

Evaluación de la litiasis renal asociada a alteraciones metabólicas / Evaluation of renal stones related to metabolic changes

Introducción. Valorar en un grupo de pacientes con litiasis renal y alteraciones metabólicas (hipercalciuria, hiperuricosuria, hipocitraturia, hiperoxaluria), cómo influye el tratamiento médico en el balance metabólico, en el número de eventos clínicos y en la necesidad de procedimientos urológicos, en seguimiento a largo plazo. Material y métodos. Estudio observacional retrospectivo. Los datos se obtuvieron de la revisión de 93 historias clínicas de pacientes que acudieron a consultas externas de medicina interna del Hospital La Princesa, Madrid, entre enero y junio de 2008. Los criterios de inclusión fueron: enfermedad litiásica renal recurrente, alteración metabólica y un mínimo de seguimiento de tres años. Se seleccionaron 68 pacientes. Resultados. Presentan hipercalciuria sola o asociada a otras alteraciones metabólicas 51/68 (75%) pacientes. Nunca habían recibido tratamiento farmacológico 57/68 (83,8%) pacientes. La necesidad de tratamiento instrumental disminuye de forma estadísticamente significativa. Se reduce el número de eventos clínicos y el balance metabólico mejora. Discusión. El tratamiento médico y el seguimiento a largo plazo favorecen la buena evolución clínica y analítica, y disminuyen la necesidad de procedimientos instrumentales (AU)

Introduction. Observational study on a group of patients with renal lithiasis and metabolic changes e.g: hypercalciuria, hyperuricosuria, hyperoxaluria, hypocitraturia. How the medical treatment would influence the restoration of the metabolic balance, the number of symptomatic events; the need for urological surgical procedures; and their long-term follow-up. Material and methods. This observational study was carried out on the data obtained from 93 medical records of patients who attended General Medicine outpatient clinics at La Princesa Hospital in Madrid between January and June 2008. The inclusion criteria were: (1) patient suffering from recurrent renal lithiasis; (2) showing metabolic changes; and (3) under survelliance for 3 or more years. Material and methods. Only 68 patients out of 93 were included in this study. Results. A total of 48/68 (70.6%) patients had hypercalciuria alone or in association with other metabolics changes. 57/68 (83.8%) patients never had appropriate pharmacological treatment. There is a statistically significant reduction in the number of cases treated by surgical procedures. There is also a reduction of clinical events and an improvement in the metabolic balance. Discussion. The medical treatment and long-term survelliance favour the good clinical and biochemical progress, and reduces the need for surgical procedures (AU)

Estudio de anticuerpos de inmunización ABO en niños con ascariosis / Study of ABO immune antibodies in children with ascariasis

Los anticuerpos inmunes tienen una gran importancia clínica, pues pueden producir reacción hemolítica o enfermedad hemolítica del recién nacido. El objetivo de este trabajo era detectar anticuerpos de inmunización ABO en niños con ascariosis. Se trabajó con muestras de suero obtenidas de una población de niños con ascariosis y de otra población control de niños sanos. Se determinó el grupo ABO en los sueros y se registró si el niño había recibido tratamiento antiparasitario en el momento de la extracción de la muestra. El estudio de los anticuerpos anti-Ay anti-B comprendió: prueba de hemólisis cualitativa, tiempo hemolítico medio, aglutinación y titulación en medio salino y enzimático, titulación en medio enzimático antes y después del tratamiento con 2-mercaptoetanol, y estudio de amplitud térmica. Se realizó también la inhibición con agarosa para los anti-B. El estudio de los anticuerpos ABO en la población de niños parasitados demostró que el 52,63% de los anti-A y el 31,03%de los anti-B tenían características de anticuerpos inmunes. No se encontró ningún anticuerpo ABO inmune en la población control. Se comprobó la existencia de 6 anticuerpos antigalactosa de clase IgG en el grupo de niños parasitados. Todos los sueros con anticuerpos inmunes fueron obtenidos antes o durante el tratamiento específico. La ausencia de anticuerpos de inmunización en los niños que concluyeron el tratamiento y en los niños de la población control sugeriría que la ascariosis es el estímulo externo para su aparición. El seguimiento de los anticuerpos inmunes sería útil para evaluar la evolución de la infección después del tratamiento (AU)

Immune antibodies are of clinical importance because they can produce a hemolytic reaction or hemolytic disease of the newborn. The aim of this study was to detect ABO immune antibodies in children with ascariasis. Serum samples were collected from a population of children with ascariasis and from a control population of healthy children. The ABO group was determined in the sera, and it was recorded if the child had received antiparasitic treatment at the time of the sample collection. The study of anti-A and anti-B antibodies involved a qualitative hemolysis test, mean hemolysis time, agglutination and titration in a saline medium and an enzyme medium, titration before and after 2- mercaptoethanol treatment in an enzyme medium, and a thermal amplitude test. An inhibition test for anti-B antibodies was also carried out in agarose. The study of ABO antibodies in the population of children with parasitemia showed that 52.63% of the anti-A and 31.03%of the anti-B antibodies had characteristics of immune antibodies. No ABO immune antibodies were found in the control population. The presence of 6 anti-galactose antibodies was demonstrated in the group with ascariasis. All the sera with immune antibodies were collected before or during the specific treatment. The absence of immune antibodies in the children that completed the treatment and in the children of the control population would suggest that ascariasis was the external stimulus for their development. The monitoring of these antibodies would be useful for the evaluation of the course of the infection following treatment (AU)

Frizzled-related protein variants are risk factors for hip osteoarthritis.

OBJECTIVE: To examine the association of the Arg200Trp and Arg324Gly variants of FRZB with the risk and phenotype of radiographic osteoarthritis (OA) of the hip and serum levels of Frizzled-related protein (FRP) in a prospective cohort of elderly Caucasian women. METHODS: Radiographic hip OA status of patients was defined by the presence of severe joint space narrowing (JSN) (feature grade>or=3), a summary grade>or=3, or definite osteophytes (grade>or=2) and JSN (grade>or=2) in the same hip. Genotypes were obtained in 569 patients with radiographic OA of the hip and in 1,317 and 4,136 controls for the Arg200Trp and Arg324Gly variants, respectively. Serum FRP levels were measured by enzyme-linked immunosorbent assay. Multivariate logistic regression was performed. RESULTS: The minor allele frequency for the Arg200Trp polymorphism was 0.12 in the control group compared with 0.14 in the group with radiographic OA of the hip (P=0.12), and the minor allele frequency for the Arg324Gly variant was 0.083 in the control group compared with 0.088 in the group with radiographic OA of the hip (P=0.63). The multilocus genotypes available in 1,886 subjects suggested that inheritance of both minor alleles was a risk factor for developing OA characterized by JSN (P<0.01). Patients with radiographic OA of the hip who were homozygous for the Arg200Trp minor allele had higher serum FRP levels than controls who were homozygous for the major allele. CONCLUSION: Our data confirm findings of another study, that a rare haplotype with both Arg200Trp and Arg324Gly FRZB variants contributes to the genetic susceptibility to hip OA among Caucasian women, and that these polymorphisms may contribute to increased serum levels of proteins as biomarkers of OA.

Ascariasis infantil: estudio de anticuerpos inmunes antigalactosa mediante glóbulos rojos desializados / Ascariasis in children: a study of anti-galactose antibodies using desialylated red blood cells

La presencia de anticuerpos inmunes del sistema ABO ha sido asociada con enfermedad hemolítica. Los anticuerpos inmunes desaparecen cuando disminuye el estímulo antigénico, por lo que su determinación cuantitativa permitiría la evaluación del curso clínico y la eficacia del tratamiento. El objetivo de este trabajo es estudiar la presencia de anticuerpos inmunes antigalactosa en sueros de niños con ascariasis. Se trabajó con 15 sueros de niños con infección por Ascaris lumbricoides. Se estudiaron los anticuerpos anti-B en el suero: actividad en medio enzimático y en medio salino; inhibición con agarosa; capacidad hemolítica y tratamiento con 2-mercaptoetanol para determinar la clase de inmunoglobulina (lg). Los anticuerpos anti-B mostraron mayor actividad en medio enzimático, presentaron capacidad hemolítica y la mitad de ellos fueron de clase IgG. Se evidenció la presencia de residuos de galactosil por inhibición con agarosa. Se demuestra la presencia de anticuerpos inmunes antigalactosa en niños con ascariasis

The presence of antibodies of the ABO system has been associated with hemolytic disease. The immune antibodies disappear when the antigenic stimulation decreases. Thus, their quantitative determination would allow the evaluation of the clinical course and treatment efficacy. The objective was to study the presence of anti-galactose antibodies in sera of children with ascariasis. Anti-B antibodies were studied in the sera of 15 chil" dren infected with Ascaris lumbricoides to assess their activity in saline and an enzymatic medium, inhibition by agarose, hemolytic capacity and treatment with 2-mercaptoethanol to determine the Ig class. Anti-B antibodies showed more activity in the enzymatic medium and presented hemolytic capacity; half of them were of the IgG class. Agarose inhibition revealed the presence of galactosyl residues. The presence of anti-galactose antibodies was demonstrated in children with ascariasis

Estudio de hemolisinas del sistema ABO por técnica fotométrica simple en niños con ascariasis / Study of hemolysins of the ABO blood group system in children with ascariasis using a simple photometric technique

La heteroinmunización en el sistema ABO, a través de sustancias de origen animal o bacteriano, puede provocar la aparición de hemolisinas de importancia clínica en transfusión o embarazo. El objetivo de este trabajo era estudiar la presencia de hemolisinas ABO en niños con ascariasis utilizando una técnica fotométrica simple. Se trabajó con sueros de 23 niños (19 antes del tratamiento antiparasitario y 4 después). Se determinó el grupo sanguíneo ABO por técnicas convencionales. La técnica fotométrica usada para demostrar la presencia de hemolisinas ABO es una modificación del tiempo hemolítico (tH) 50. El tiempo hemolítico medido tiene significado clínico cuando es inferior a 300 segundos. El 57,89% de los sueros de los niños sin tratamiento presentó hemolisinas ABO, con tiempos hemolíticos comprendidos entre 100 y 210 segundos. Ninguno de los sueros de los niños tratados presentó hemolisinas ABO. Los resultados sugieren que la infección parasitaria puede ser el estímulo externo para la aparición de hemolisinas ABO. La técnica fotométrica usada es simple, rápida y accesible al laboratorio de rutina

Heteroimmunization in the ABO system by animal or bacterial products can provoke the development of hemolysins of clinical importance in transfusions or pregnancy. We proposed to study the presence of ABO hemolysins in children with ascariasis using a simple photometric technique. Serum samples were collected from 23 children (19 prior to and 4 after treatment with antiparasitic agents). The ABO blood group was determined by conventional techniques. The photometric technique employed to demonstrate the presence of ABO hemolysins is a modification of tH 50. A hemolysis time of less than 300 seconds was considered to be clinically significant. We detected ABO hemolysins in 57.89% of the sera of untreated children, with hemolysis times ranging between 100 and 210 seconds. None of the sera from the treated children presented ABO hemolysins. The results suggest that parasitic infection may be the external stimulus that provokes the appearance of ABO hemolysins. The photometric technique employed is simple and rapid, and is available to any routine laboratory

"Giardia lamblia" en una población infantil / "Giardia lamblia" in a pediatric population

Se estudiaron 8.717 niños menores de 13 años de edad, de ambos sexos durante los años 1992-1996. Objetivos: determinar la frecuencia de Giardia lamblia e una población infantil, la distribución de este agente etiológico de acuerdo a la edad de los niños parasitados y la variación de la frecuencia de la infección en los meses del año. Material y métodos: a todos los pacientes se les solicitó una muestra de heces recogida de forma seriada durante 8 días consecutivos conservadas en formol al 10 por ciento. Las heces se examinaron por microscopía directa en X100 y X400 aumentos para la búsqueda de trofozoítos y/o quistes de G. lamblia. Resultados: de las 8.717 muestras de materia fecal estudiadas 2.290 (26,27 por ciento) presentaron G. lamblia. Los porcentajes de infección variaron del 22,78 al 32,30 por ciento, según la edad, mostrando un pico máximo entre los 3 y 4 años, y del 18,36 al 21,74 por ciento en los menores de 1 año y 12 años, respectivamente. no se observó tendencia en las variaciones mensuales. Conclusiones: el presente estudio sugiere que G. lamblia constituye un patógeno de alta prevalencia en niños, con una frecuencia similar a la comunicada por otros autore, lo que hace necesario su diagnóstico para la aplicación de una terapia adecuada (AU)

Leaves of Cercocarpus mixteca n. sp. (Rosaceae) from Oligocene sediments, near Tepexi de Rodríguez, Puebla.

A new species of Cercocarpus, Cercocarpus mixteca Velasco de León & Cevallos-Ferriz, is described based on leaf impressions from the Los Ahuehuetes locality, near Tepexi de Rodríguez, Puebla, Mexico. The lamina is obovate, 1.3cm in length by 0.5cm in width, has a serrate margin in its distal fourth, craspedodromous venation with a single straight mid-vein and two to four pairs of secondary ones, and areols that tend to be quadrangular in shape. A phenetic analysis of the agglomerative, non-hierarchical type, with mean linkage, is applied using 22 OTUs and 34 character states. The morphological characters observed on the leaves of the new fossil plants support the recognition of a new taxon closely related to the extant Cercocarpus paucidentatus growing naturally in northern Mexico. Its microphyll size corresponds with the temperate to xeric climate postulated for the Los Ahuehuetes locality; this further suggests that some taxa, like Cercocarpus, have a long history in low latitude North America. In this particular case, the extant Cercocarpus fothergilloides and Cercocarpus macrophyllus could, as they were able to colonise new humid and xeric areas, represent descendants of C. mixteca.

Large-scale purification of antisense oligonucleotides by high-performance membrane adsorber chromatography.

Very high flux ion-exchange membranes were utilized for a novel purification of antisense oligonucleotides (20-mer). Strong anion-exchange membranes were produced by attaching polymeric ligands onto a microporous cellulosic matrix. The oligonucleotides purified were therapeutic single-stranded phosphorothioates deoxyribonucleotides. Although small-scale membrane devices (15 cm2) had similar resolution to traditional chromatographic columns; their throughputs were superior. Greater than a 1300-fold scale-up produced very similar purity and yields of the phosphorothionate product. Scale-up experiments were conducted with a 2 m2 surface area membrane module. These modules were easily capable of very high throughputs of 0.5 to 2 l/min. High purity and yields were achieved by both step and linear gradient elution.

Genomic variability of Haemophilus influenzae isolated from Mexican children determined by using enterobacterial repetitive intergenic consensus sequences and PCR.

Genomic fingerprints from 92 capsulated and noncapsulated strains of Haemophilus influenzae from Mexican children with different diseases and healthy carriers were generated by PCR using the enterobacterial repetitive intergenic consensus (ERIC) sequences. A cluster analysis by the unweighted pair-group method with arithmetic averages based on the overall similarity as estimated from the characteristics of the genomic fingerprints, was conducted to group the strains. A total of 69 fingerprint patterns were detected in the H. influenzae strains. Isolates from patients with different diseases were represented by a variety of patterns, which clustered into two major groups. Of the 37 strains isolated from cases of meningitis, 24 shared patterns and were clustered into five groups within a similarity level of 1.0. One fragment of 1.25 kb was common to all meningitis strains. H. influenzae strains from healthy carriers presented fingerprint patterns different from those found in strains from sick children. Isolates from healthy individuals were more variable and were distributed differently from those from patients. The results show that ERIC-PCR provides a powerful tool for the determination of the distinctive pathogenicity potentials of H. influenzae strains and encourage its use for molecular epidemiology investigations.