OM-85 BV for primary prevention of recurrent airway infections: a pilot randomized, double-blind, placebo-controlled study / OM-85 BV para prevenção primária de infecções recorrentes das vias aéreas: estudo piloto, randomizado, duplo-cego, placebo-controlado

ABSTRACT Objective To compare the frequency of respiratory tract infections in children treated with OM-85 BV and placebo during the 3-month therapy period, and observation for a further 3 months after treatment. Methods A randomized, double-blind, placebo-controlled trial was conducted with 54 children (6 months to 5 years old) with no past history of recurrent respiratory infections attending daycare center. Family members were instructed to administer one capsule per day for 10 consecutive days, for 3 months of OM-85 BV or placebo. Telephone interviews were conducted every 30 days. Results There was no significant difference in the number of respiratory infections between the groups. The mean number of respiratory tract infection in the OM-85 BV Group in the first 3 months was 0.92±0.87, and in the Placebo Group was 0.74±1.02, and at 6 months it was 1.62±1.47 and 1.03±1.34, respectively. Conclusion OM-85 BV was not effective in the primary prevention of respiratory tract infections. Although most authors recommend the use of this immunostimulant in children with a history of recurrent respiratory infections, more studies are needed to define its usefulness in the primary prevention of respiratory infections in healthy children exposed to few risk factors.

RESUMO Objetivo Comparar a frequência de infecções do trato respiratório em crianças tratadas com OM-85 BV e placebo durante o período de terapia de 3 meses, e observação por mais 3 meses após o tratamento. Métodos Foi realizado estudo randomizado, duplo-cego, controlado por placebo com 54 crianças (6 meses a 5 anos) sem história prévia de infecções respiratórias recorrentes, que frequentavam creches. Os membros da família foram instruídos a administrar uma cápsula por dia durante 10 dias consecutivos, durante 3 meses, de OM-85 BV ou placebo. Entrevistas telefônicas foram realizadas a cada 30 dias. Resultados Não houve diferença significativa no número de infecções respiratórias entre os grupos. O número médio de infecções do trato respiratório no Grupo OM-85 BV nos primeiros 3 meses foi de 0,92±0,87 e, no Grupo Placebo, de 0,74±1,02, e aos 6 meses foi de 1,62±1,47 e 1,03±1,34, respectivamente. Conclusão O OM-85 BV não foi eficaz na prevenção primária de infecções do trato respiratório. Embora a maioria dos autores recomende o uso deste imunoestimulante em crianças com história de infecções respiratórias recorrentes, mais estudos são necessários para definir sua utilidade na prevenção primária de infecções respiratórias em crianças saudáveis expostas a poucos fatores de risco.

Utilization of a waste glycerol fraction using and reusing immobilized Gluconobacter oxydans ATCC 621 cell extract

Background: Depletion of petroleum resources has enforced the search for alternative sources of renewable energy. Introduction of biofuels into the market was expected to become a solution to this disadvantageous situation. Attempts to cover fuel demand have, however, caused another severe problem­the waste glycerol generated during biodiesel production at a concentration of approximately 10% w/w. This, in turn, prompted a global search for effective methods of valorization of the waste fraction of glycerol. Results: Utilization of the waste fraction at 48 h with an initial glycerol concentration of 30 g·L-1 and proceeding with 62% efficiency enabled the production of 9 g·L-1 dihydroxyacetone at 50% substrate consumption. The re-use of the immobilized biocatalyst resulted in a similar concentration of dihydroxyacetone (8.7 g·L-1) in two-fold shorter time, with an efficiency of 85% and lower substrate consumption (35%). Conclusions: The method proposed in this work is based on the conversion of waste glycerol to dihydroxyacetone in a reaction catalyzed by immobilized Gluconobacter oxydans cell extract with glycerol dehydrogenase activity, and it could be an effective way to convert waste glycerol into a valuable product.

Homocysteine induces glyceraldehyde-3-phosphate dehydrogenase acetylation and apoptosis in the neuroblastoma cell line Neuro2a

High plasma levels of homocysteine (Hcy) promote the progression of neurodegenerative diseases. However, the mechanism by which Hcy mediates neurotoxicity has not been elucidated. We observed that upon incubation with Hcy, the viability of a neuroblastoma cell line Neuro2a declined in a dose-dependent manner, and apoptosis was induced within 48 h. The median effective concentration (EC50) of Hcy was approximately 5 mM. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) nuclear translocation and acylation has been implicated in the regulation of apoptosis. We found that nuclear translocation and acetylation of GAPDH increased in the presence of 5 mM Hcy and that higher levels of acetyltransferase p300/CBP were detected in Neuro2a cells. These findings implicate the involvement of GAPDH in the mechanism whereby Hcy induces apoptosis in neurons. This study highlights a potentially important pathway in neurodegenerative disorders, and a novel target pathway for neuroprotective therapy.

Silver nanoparticle production by the fungus Fusarium oxysporum: nanoparticle characterisation and analysis of antifungal activity against pathogenic yeasts

The microbial synthesis of nanoparticles is a green chemistry approach that combines nanotechnology and microbial biotechnology. The aim of this study was to obtain silver nanoparticles (SNPs) using aqueous extract from the filamentous fungus Fusarium oxysporum as an alternative to chemical procedures and to evaluate its antifungal activity. SNPs production increased in a concentration-dependent way up to 1 mM silver nitrate until 30 days of reaction. Monodispersed and spherical SNPs were predominantly produced. After 60 days, it was possible to observe degenerated SNPs with in additional needle morphology. The SNPs showed a high antifungal activity against Candida and Cryptococcus , with minimum inhibitory concentration values ≤ 1.68 µg/mL for both genera. Morphological alterations of Cryptococcus neoformans treated with SNPs were observed such as disruption of the cell wall and cytoplasmic membrane and lost of the cytoplasm content. This work revealed that SNPs can be easily produced by F. oxysporum aqueous extracts and may be a feasible, low-cost, environmentally friendly method for generating stable and uniformly sized SNPs. Finally, we have demonstrated that these SNPs are active against pathogenic fungi, such as Candida and Cryptococcus .

Assessment of the phytochemical constituents and antioxidant activity of a bloom forming microalgae Euglena tuba

BACKGROUND: Unstable generation of free radicals in the body are responsible for many degenerative diseases. A bloom forming algae Euglena tuba growing abundantly in the aquatic habitats of Cachar district in the state of Assam in North-East India was analysed for its phytochemical contents, antioxidant activity as well as free radical scavenging potentials. RESULTS: Based on the ability of the extract in ABTS•+ radical cation inhibition and Fe3+ reducing power, the obtained results revealed the prominent antioxidant activity of the algae, with high correlation coefficient of its TEAC values to the respective phenolic and flavonoid contents. The extract had shown its scavenging activity for different free radicals and 41.89 ± 0.41 µg/ml, 5.83 ± 0.07 µg/ml, 278.46 ± 15.02 µg/ml and 223.25 ± 4.19 µg/ml were determined as the IC50 values for hydroxyl, superoxide, nitric oxide and hypochlorous acid respectively, which are lower than that of the corresponding reference standards. The phytochemical analysis also revealed that the phenolics, flavonoids, alkaloids, tannins and carbohydrates are present in adequate amount in the extract which was confirmed by HPLC analysis. CONCLUSIONS: The results showed that 70% methanol extract of the algae possesses excellent antioxidant and free radical scavenging properties.

TAPCells, the Chilean dendritic cell vaccine against melanoma and prostate cancer

Here we summarize 10 years of effort in the development of a biomedical innovation with global projections. This innovation consists of a novel method for the production of therapeutic dendritic-like cells called Tumor Antigen Presenting Cells (TAPCells®). TAPCells-based immunotherapy was tested in more than 120 stage III and IV melanoma patients and 20 castration-resistant prostate cancer patients in a series of phase I and I/II clinical trials. TAPCells vaccines induced T cell-mediated memory immune responses that correlated with increased survival in melanoma patients and prolonged prostate-specific antigen doubling time in prostate cancer patients. Importantly, more than 60% of tested patients showed a Delayed Type Hypersensitivity (DTH) reaction against the lysates, indicating the development of anti-tumor immunological memory that correlates with clinical benefits. The in vitro analysis of the lysate mix showed that it contains damage-associated molecular patterns such as HMBG-1 protein which are capable to improve, through Toll-like receptor-4, maturation and antigen cross-presentation of the dendritic cells (DC). In fact, a Toll-like receptor-4 polymorphism correlates with patient clinical outcomes. Moreover, Concholepas concholepas hemocyanin (CCH) used as adjuvant proved to be safe and capable of enhancing the immunological response. Furthermore, we observed that DC vaccination resulted in a three-fold increase of T helper-1 lymphocytes releasing IFN-γ and a two-fold increase of T helper-17 lymphocytes capable of producing IL-17 in DTH+ with respect to DTH- patients. Important steps have been accomplished for TAPCells technology transfer, including patenting, packaging and technology assessment. Altogether, our results indicate that TAPCells vaccines constitute an exceptional Chilean national innovation of international value.

Diferencias proteómicas entre una cepa argentina de Mycobacterium bovis con la cepa de referencia AN5 / Proteomic differences between an argentine mycobacterium bovis strain and the reference AN5 strain

La tuberculosis bovina es en Argentina una enfermedad que provoca graves pérdidas económicas y que afecta a un 5 por ciento del ganado. En un trabajo previo de tipificación de cepas por RFLP mediante el uso de las sondas PGRS y DR se identificó una cepa altamente predominante que se llamó AA. A diferencia de la cepa salvaje AA, la cepa de referencia AN5, de origen europeo, que se utiliza para elaborar la tuberculina, es una cepa adaptada al crecimiento en laboratorio, que puede haber sufrido mutaciones en genes de antígenos o de virulencia. Para ello se analizó la producción de proteínas secretadas y del extracto celular, de la cepa salvaje AA comparada con la cepa de referencia AN5, con el propósito de identificar diferencias que puedan dar cuenta de la virulencia y para identificar nuevos antígenos. Se utilizaron técnicas como electroforesis en geles de policrilamida, geles de 2 dimensiones y Western blot utilizando antisueros específicos contra antígenos ya caracterizados y sueros de bovinos infectados con tuberculosis confirmada por aislamiento de M. bovis, empleando proteínas celulares y secretadas (a los 25 y 100 días de cultivo) de ambas cepas. Se pudieron identificar, una proteína secretada de aproximadamente 29 kDa y otra de 28 kDa del estracto celular que parecen ser exclusivas o producidas en mayor cantidad por la cepa AA. También, se identificaron otras pero cuyas bandas eran más débiles. En conclusión, algunas de las proteínas identificadas pueden servir para mejorar el diagnóstico de la tuberculosis bovina

Glicoprotéinas de Klebsiella pneumoniae como coadyuvantes en la prevención de infecciones recurrentes respiratorias en niños de 1 a 6 años de edad / The extract of Klebsiella pneumoniae can be used as a coadyuvant in the treatment of pediatric patients with recurrent respiratory infections

Antecedentes. Las infecciones recurrentes de las vías respiratorias altas y bajas representan los padecimientos más comunes de la edad pediátrica. Material y método. Estudio doble ciego, pareado y en paralelo. Se comparó la mejoría clínica y de laboratorio con un grupo de 44 pacientes control, a quienes se administró placebo, con un diagnóstico semejante, lo mismo que edades y distribución por sexo. Resultados. En el grupo de estudio, comparado con el de control (p<0.001 en todos los parámetros), a los 6, 9 y 12 meses posteriores al inicio del tratamiento hubo una disminución estadísticamente significativa del número de infecciones por año, duración de éstas y días de ausencia escolar. No hubo cambios estadísticamente significativos en las concentraciones de complemento hemolítico, C3, C4 e inmunoglobulinas IgA, IgG, IgM e IgE, cuando se compararon los valores basales y a los seis meses de iniciado el tratamiento. No se reportó reacción adversa a la administración oral de la glicoproteína de Klebsiella pneumoniae. Conclusión. Estos resultados sugieren que ésta puede usarse como coadyuvante en la prevención y tratamiento de pacientes con infecciones respiratorias bajas y altas recurrentes

Expression of the Mycobacterium bovis P36 gene in Mycobacterium smegmatis and the baculovirus/insect cell system

In the present study we evaluated different systems for the expression of mycobacterial antigen P36 secreted by Mycobacterium bovis. P36 was detected by Western blot using a specific antiserum. The P36 gene was initially expressed in E. coli, under the control of the T7 promoter, but severe proteolysis prevented its purification. We then tried to express P36 in M. smegmatis and insect cells. For M. smegmatis, we used three different plasmid vectors differing in copy number and in the presence of a promoter for expression of heterologous proteins. P36 was detected in the cell extract and culture supernatant in both expression systems and was recognized by sera from M. bovis-infected cattle. To compare the expression level and compartmentalization, the MPB70 antigen was also expressed. The highest production was reached in insect cell supernatants. In conclusion, M. smegmatis and especially the baculovirus expression system are good choices for the production of proteins from pathogenic mycobacteria for the development of mycobacterial vaccines and diagnostic reagents

Efecto in vitro del extracto bacteriano de S. aureus en la quimiocinesis y quimiotaxis de células polimorfonucleares / Effect the extract of Staphylococcus aureus in vitro can modify chemokinesis and induce chemotaxis of polimorphonuclear cells

El propósito del trabajo fue determinar si el extracto de Staphylococcus aureus in vitro puede modificar la quimiocinesis e inducir la quimiotaxis de las células polimorfonucleares de la sangre periférica en donadores sanos. Se determinó la quimiocinesis y la quimiotaxis de los polimorfonucleares de la sangre periférica de 30 donadores sanos de uno y otro sexo con un límite de edad entre 18 y 40 años. Se les extrajeron 5 mL de sangre periférica separando los polimorfonucleares por el método de Boyum y se retaron con extracto de Staphylococcus aureus y C5a como quimiotácticos, y solución de Hank para medir quimiocinesis. Esta tuvo un promedio de 54.6ñ8.8 µm, la respuesta quimiotáctica a C5a fue 89ñ12.5 µm y con el extracto bacteriano fue 103ñ20.1 µm (p<0.001. Análisis estadístico: prueba de Wilcoxon. Se concluye que el extracto completo de Staphylococcus aureus estímula in vitro la quimiotaxis de polimorfonucleares de donadores sanos y que esta estimulación es comparable con lo quimioatrayentes conocidos como C5a

Apertura palpebral prematura experimental y desarrollo neuronal en la rata recién nacida / Experimental premature eyelid openingand neuron development in new-born rats

Los extractos de glándula submaxilar (EGS) de ratón induce apertura prematura de los párpados en ratas recién nacidas. En animales sometidos a ciclos alternos de luz-obscuridad y con EGS, las neuronas de la corteza visual exhiben, en la dendrita principal, más espinas y un mayor grado de ramificación que los controles, en tanto que en animales mantenidos en obscuridad total y con EGS, únicamente disminuyen las espinas dendríticas, mientras que el grado de ramificación es mayor comparado con los controles.

Virulence of Candida albicans "clones"

Com base em características morfo-fisiológicas e cromossomais foram selecionados 5 clones oriundos de diferentes cepas de Candida albicans. Por meio de inoculaçöes em camundongos investigou-se a atividade letal de blastósporos e de extratos intra e extracelulares das fases leveduriforme (Y) e micelial (M) dos 5 clones. A inoculaçäo de 5.10(4ª) blastósporos de qualquer dos clones estudados näo foi letal para os animais. Por outro lado, a inoculaçäo de 10(6ª) mostrou que o clone Ca 3.7 foram letais para todos os animais, logo após a inoculaçäo. Entretanto, o extrato extracelular da forma M deste clone näo mostrou toxicidade. A análise dos extratos oriundos do clone Ca 5.5 näo apresentaramtoxicidade. A análise dos extratos dos diferentes clones por SDS-PAGE näo mostrou ocorrência de uma nítida associaçäo entre virulência dos clones e a toxicidade dos extratos. A partir do estudo com macrófagos "in vitro" obteve-se para o clone Ca 5.5 o maior índice de fagocitose. Entretanto, a percentagem de macrófagos com células mortas foi a mesma para os clones Ca 3.7 e Ca 5.5

Solubilización de la membrana de espermatozoide humano con n-dodecil sarcosinato de sodio: II. Estudios sobre el extracto / Spermatozoa submitted to ionic detergent treatment: studen on the sodium n-dodceyl sarcosinate extract

El n-dodecil sarcosinato de sodio produce la solubilización del espermatozoide humano con algo de contaminación acrosomal. El extracto está compuesto por una mezcla de productos con pesos moleculares y características inmunológicas similares a las del plasma seminal humano. El hecho de que la antigenicidad se mantenga luego de la solubilización, hace que ésta sea un comienzo útil en el aislamiento de moléculas que juegan un papel en la infertilidad por causas inmunológicas

The immunodominant surface antigen of Plasmodium gallinaceum is present in both the salivary gland and oocyst sporozoites

1. Monoclonal antibodies (MAbs) against surface antigens of Plasmodium gallinaceum sporozoites, an avian malaria parasite, were produced using spleen cells from mice immunized with sporozoites from mosquito salivary glands (SGS) or from midadgusts containing oocysts (OoS). 2. All of the 15 MAbs teted (11 anti-SGS and 4 anti-OoS) reacted with SGS and OoS by indirect immunofluorescence and circumsporozoiter precipitation reactions. Fourteen of these MAbs (11 anti-SGS and 3 anti-OoS) produced a Western blot (WB) patten identical to that produced with serum from mice lyperimmunized with viable intacts sporozoites. 3. All MAbs and the immune sera recognized only two polypeptide bands of approximate molecutlar weight 78 and 64KDa. 4. No difference in the WB pattern was observed when-or 12-day SGS or OoS extracts were used as antigens in WB. This antigenic similary was confirmed when the total protein extracts were visualized on silver-stained SDS-PAGE gel

Acción del extracto leucocitario dializado sobre la maduración de timocitos in vitro / Action of dialyzed leukocyte extract on in vitro maturation of thymocytes

Basado en las sugerencias tempranas de Lawrence, acerca de que los efectos inmunológicos específicos y no específicos del extracto leucocitario dializado (ELD) puedan derivar de su capacidad de iniciar la maduración de una célula primitiva de la serie T, se estudiaron los efectos del ELD sobre la adquisición de la capacidad de respuesta blastogénica de los timocitos de ratón a los fitomitógenos: PHA, con A y PWM y sobre la conversión de los timocitos hidrocortisona-sensibles en hidrocortisona-resistentes como indicadores reconocidos de la maduración de células T. Nuestros resultados destacan el ELD como posible inductor de la maduración y diferenciación terminal de las células tímicas, propiedad exclusiva del ELD respecto a otros inmunosupresores y fuente de importantes implicaciones terapéuticas