Dermal toxicity, eye and dermal irritation and skin sensitization evaluation of a new formulation of Bacillus thuringiensis var israelensis SH-14.

Bacillus thuringiensis (Bt) is the best known and most widely used of all pesticidal microbes. The aim of this study was to assess the toxicity of a new formulation of Bacillus thuringiensis var israelensis SH-14 in rats through acute dermal toxicity, dermal and eye irritation experiments. The acute dermal toxicity and dermal and eye irritation studies were performed using rabbits according to the United States Environmental Protection Agency guidelines 885.3100, 870.2500 and 870.2500, respectively. The skin sensitization study was carried out in accordance to the EPA OPPTS 870.2600 using guinea pigs. There was no mortality and no evidence of treatment-related toxicity in acute dermal toxicity test. No dermal responses, including erythema/eschar or edema, were found in rabbits treated with the new formulation of Bti SH-14. Minimum response was observed after eye application of test substance. No skin sensitization reactions were observed after the challenge with the new formulation of Bti SH-14 in the Bti SH-14-treated guinea pigs. In summary, the present study demonstrated that the new formulation of Bti SH-14 is not acutely toxic via dermal route, has low eye irritation and would not cause dermal irritation or hypersensitivity to tested animals.

Repeated dose (14 days) rat intramuscular toxicology study of Her1 vaccine.

Our goal was to assess the toxicity of two strengths (200 and 400 µg) of HER1 cancer vaccine (Center of Molecular Immunology, Cuba), presented in two different formulations, in Sprague Dawley rats after repeated intramuscular administration (14 days). Four groups (5 animals/sex) were established: Control, Placebo (adjuvant), and two Treated groups receiving a dose representing ten times of human total dose (10×), 28.6 and 57.1 µg/kg. Clinical observations, body weight and rectal temperature were measured during the study. Clinical pathology analysis was performed, besides gross necropsy and histological examination of tissues on animals at the end of the assay. The assay ended with a 100% survival. Injection site damage, with the presence of cysts and granulomas, was observed in adjuvant and vaccine treated groups, with most severe cases predominating at higher strength. Administration of Placebo and Her1 vaccine induced increase in polymorphonuclear cells, with relative lymphopenia conditioned by primary neutrophilia. In summary, results suggest that Her1 immunization was capable of inducing an inflammatory effect at the injection site, leading to systemic alterations, more significant at higher strength (400 µg, 57.1 µg/kg), probably affected by the immunizations' schedule used. The vaccine was shown to be well tolerated without any obvious signs of systemic toxicity, with findings largely attributable to the adjuvant used.

Pregnancy after percutaneous epididymal sperm aspiration in an 81-year-old man with obstructive azoospermia.

There has been observed a trend to delay childbearing, reflecting couple's desire to have children at older ages. Maternal age is a well-known factor that influences the achievement of a pregnancy. In contrast, there are few studies examining the effect of paternal age on the outcomes of assisted reproductive technologies (ART), and results are conflicting. Our patient was vasectomised at the age of 60 years, and his wife was 38 years old. A total of four metaphase II oocytes were inseminated 4 h later (day 0) by intracytoplasmic sperm injection using spermatozoa from percutaneous epididymal sperm aspiration. On day 2, the three resulting embryos were transferred (two had four cells, and one had two cells). Two healthy infants were born at the 37th gestational week by caesarean section. Although the effect of paternal age on ART outcomes and results are conflicting, this case report contributes to point out that advanced age of the man has probably lesser negative influence in the reproductive function.

Repeated dose intramuscular injection of the CIMAvax-EGF vaccine in Sprague Dawley rats induces local and systemic toxicity.

CIMAvax-EGF consists of a human recombinant epidermal growth factor (EGF), coupled to P64k, a recombinant carrier protein from N. meningitis, and Montanide ISA 51 as adjuvant. The vaccine immunization induces a specific antibody production, inhibiting the EGF/EGF-R interaction through EGF deprivation. The objective of this study was to assess the CIMAvax-EGF toxicity in Sprague Dawley rats after intramuscular administration of repeated doses (6 months) and at the same time to determine if rat is a relevant species for studying CIMAvax-EGF vaccine. Rats were randomly distributed into four groups: control, Montanide ISA 51, treated with 1× and 15× of human total dose of the antigen. Animals were immunized weekly during 9 weeks, plus 9 immunizations every 14 days. Rats were inspected daily for clinical signs. Body weight, food consumption, and rectal temperature were measured during the administration of doses. Blood samples were collected for hematological, serum biochemical determinations and EGF titles at the beginning, three months and at the end of experimentation. Gross necropsy and histological examination of tissues were performed on animals at the end of the assay. Vaccine provoked the apparition of antibodies against EGF in the rats, demonstrating rat species relevance in these studies. Body weight gain, food and water consumption were not affected. CIMAvax-EGF and Montanide ISA 51 produced local damage at the administration site, showing multiple cysts and granulomas. Both vaccine-treated groups showed neutrophil elevation, besides an AST increase probably related to the damage at the administration site. Rectal temperature was found to be significantly higher in 15× treated group after immunizations, probably induced by the inflammatory process at the injection site. In summary, the clinical pathology findings together with the body temperature results, appear to be caused by the inflammatory reaction at the administration site of the vaccine, mainly mediated by the oil-based adjuvant Montanide ISA 51, probably enhanced by the immunological properties of the antigen. This study showed evidences that intramuscular administration during 26 weeks of CIMAvax-EGF at doses up to 15× human total dose is well tolerated in rats and it has a clinical importance since this long lasting study in relevant species allows to treat cancer patients with tumors during long periods with relative weight safety margin.

Acute oral, pulmonary and intravenous toxicity/pathogenicity testing of a new formulation of Bacillus thuringiensis var israelensis SH-14 in rats.

During the last decades, efforts are being made to develop microbial insecticides as biological control agents. Bacillus thuringiensis has been one of the most consistent and significant biopesticides for using on crops as an insecticidal spray. The aim of this study was to assess and to compare the pathogenicity of a new formulation of B.thuringiensis var israelensis SH-14 in rats through oral, intranasal and intravenous single dosing. Through 21 days after administration, clinical examinations were performed daily, and body weight gain was evaluated. Clearance was estimated by means of collection of feces or examination of lungs and blood, and infectivity was evaluated enumerating microorganisms from organs of Bti SH-14 treated animals sacrificed at intervals. Gross necropsy of animals was performed at interim or final sacrifice. There were no treatment-related mortalities, and no evidence of pathogenicity or treatment related toxicity, although in the intravenous study, the microorganism was capable of achieving persistence in organs after administration, and the Bti SH-14 treated animals developed skin ulcerations and hemorrhages at the injection site. It could be concluded that the tested microorganism was not toxic or pathogenic to rats via oral or intranasal route, although it was capable of achieving persistence in organs after intravenous administration, eliciting local effects at the injection site.

Indolent evolution of an episode of acute gastroenteritis complicated with portal pneumatosis.

The discovery of gas in the portal system related to indolent clinical settings has increased in recent years due to a growth in the performance of imaging tests. We report the first case of spontaneous resolution of portal pneumatosis due to acute gastroenteritis described in the literature.

Resolución espontánea de neumatosis portal tras un episodio de gastroenteritis aguda / No disponible

El hallazgo de gas en el sistema portal en relación a cuadros de evolución indolente ha aumentado en los últimos años por la mayor realización de pruebas de imagen. Creemos importante destacar que, a pesar de los hallazgos radiológicos, el sentido clínico es lo más importante a la hora de enfocar el diagnóstico. Presentamos el primer caso en la literatura de neumatosis portal secundaria a un cuadro de gastroenteritis aguda, con resolución espontánea(AU)

The discovery of gas in the portal system related to indolent clinical settings has increased in recent years due to a growth in the performance of imaging tests. We report the first case of spontaneous resolution of portal pneumatosis due to acute gastroenteritis described in the literature(AU)

Free radical production from the interaction of 2-chloroethyl vesicants (mustard gas) with pyridine nucleotide-driven flavoprotein electron transport systems.

The biochemical sequelae to chloroethyl mustard exposure correspond very well to toxic processes initiated by free radicals. Additionally, mustard solutions contain spontaneously formed cyclic onium ions which produce carbon free radicals when reduced electrochemically. Therefore, we hypothesized that the onium ions of sulfur or nitrogen mustards might produce carbon free radicals upon being reduced enzymatically, and that these radicals might constitute a metabolic activation. We set out to document radical production using an in vitro metabolic system and electron paramagnetic resonance (EPR). Our system consisted of NADPH, one of several pyridine nucleotide-driven flavoprotein reductases, cytochrome c as a terminal electron acceptor, various sulfur or nitrogen mustards and the spin trap alpha-[4-pyridyl-1-oxide]-N-tert-butylnitrone in buffer. Reactions were started by adding the reductase to the other materials, vortexing and immediately transferring the mixture to a 10 mm EPR flat cell. Repeated scans on a Bruker ESP 300E EPR spectrometer produced a triplet of doublets with hyperfine splitting constants of a(N)=15.483 G and a(H)=2.512 G. The outcome supported our hypothesis that carbon-centered free radicals are produced when mustard-related onium ions are enzymatically reduced. The EPR results varied little with the chloroethyl compound used or with porcine or human cytochrome P450 reductase, the reductase domain of rat brain neuronal nitric oxide synthase or rat liver thioredoxin reductase. Our results offer new insight into the basis for mustard-induced vesication and the outcome of exposure to different mustards. The free radical model provides an explanation for similarities in the lesions arising from mustard exposure and energy-based lesions such as those from heat, ultraviolet and nuclear radiation as well as damage across tissue types such as skin, eyes or airway epithelium.

Evaluación de la patogenicidad en ratas del Paecilomyces lilacinus LPL-01 utilizando vías diferentes de exposición / Pathogenicity evaluation of Paecilomyces lilacinus to rats

Los nemátodos parásitos de las plantas, conocidos como plagas agrícolas desde el siglo XIX, causan un 9% de pérdidas de cultivos en los países desarrollados y un 14% en los países en desarrollo. El Paecilomyces lilacinus es un hongo parásito que ataca formas sedentarias de los nemátodos, como los huevos. Su valoración como agente microbiano de control debe incluir una evaluación de su virulencia hacia organismos no-diana, tomando en consideración las vías posibles de exposición de los humanos. Para evaluar la patogenicidad de la cepa LPL-01 del P. lilacinus en ratas, se administró por las vías oral, pulmonar e intravenosa. Las observaciones clínicas fueron diarias, y se evaluó el comportamiento del peso corporal. Se estimó el aclaramiento mediante recolección de las heces fecales y análisis de muestras de los pulmones y de la sangre, según la vía de administración, y se evaluó la infectividad mediante toma de muestras de órganos de animales inoculados sacrificados a intervalos. Durante estos sacrificios, y al final de los ensayos, se realizó la necropsia de los animales. No ocurrieron mortalidades, ni evidencias de patogenicidad relacionada con el tratamiento en los ensayos oral y pulmonar, no provocando el hongo una infestación significativa. Por vía endovenosa, el microorganismo provocó alteraciones anátomo-patológicas en hígado y bazo, coincidiendo con el período de máxima infestación. Se concluyó que la cepa LPL-01 del P. lilacinus, a las dosis evaluadas, no es patogénica por las vías oral y pulmonar, siendo levemente patogénica por vía endovenosa

Plant parasitic nematodes have been recognized as agricultural pests in Europe as early as the late 19th century. It has been estimated that plant parasitic nematodes cause crop yield losses of nearly 9% in the developed world, and over 14% in developing countries. The Paecilomyces lilacinus is a parasitic fungi attacking sedentary stages of nematodes, e.g. eggs. Evaluation of this fungus as a microbial control agent, must include an evaluation of its virulence towards non-target organisms, especially vertebrates, with consideration given to potential human exposure scenarios. With the aim of assessing the pathogenicity in rats of the strain LPL-01 of Paecilomyces lilacinus, this fungus was given using several routes of exposure (oral, pulmonary and intravenous route). In all of the assays, clinical examinations were performed daily after administration, and body weight gain of animals was evaluated. Clearance was estimated by means of collection of feces and examination of lungs and blood, depending on the route used, and ineffectiveness was evaluated by enumerating microorganisms from organs and corporal fluids in animals sacrifice at intervals. A gross necropsy of all animals was performed at interim or final sacrifice. There were no mortalities, and no evidence of pathogenicity or treatment-related toxicity either in oral or pulmonary toxicity/pathogenicity tests, without significant infection of test animals. In the intravenous toxicity/pathogenicity test, P. lilacinus caused anatomopathological changes in liver and spleen at the same period when higher infectivity was achieved. It was concluded that P. lilacinus is not pathogenic by oral and pulmonary route, but has some pathogenic effects when intravenous injection is performed

Evaluación de la toxicidad/patogenicidad de una formulación de Bacillus thuringiensis var israelensis (Bactivec) / Toxicity/pathogenicity evaluation of a Bacillus sphaericus 2362 formulation (Griselesf)

Existen varios agentes microbianos, incluyendo hongos, protozoos, virus y bacterias, que han sido utilizados como mosquiticidas. No obstante, entre esos agentes, el Bacillus thuringiensis es el más potente y ha sido el más ampliamente utilizado. El Bactivec es un biolarvicida producido en Cuba, que contiene Bacillus thuringiensis variedad israelensis como agente activo. Con el objetivo de evaluar su toxicidad/patogenicidad, se administró el Bactivec en una dosis única de 5 x 108 unidades formadoras de colonias por vía oral a ratas, y por vía dérmica a dos grupos de conejos albinos, uno recibiendo una dosis de Bactivec de l x 109 unidades formadoras de colonia por animal, y el otro una dosis de 9.2 x 108 de unidades formadoras de colonia de Bacillus thuringiensis variedad israelensis por animal. En ambos ensayos las observaciones clínicas fueron diarias, y se evaluó el comportamiento del peso corporal. Además, en el ensayo por vía oral se estimó el aclaramiento mediante recolección de las heces fecales, y se evaluó la infectividad mediante toma de muestras de fluidos y órganos. Al final de los ensayos se realizó la necropsia ä todos los animales. No ocurrieron mortalidades, ni evidencias de patogenicidad o toxicidad relacionada con el tratamiento en ninguno de los ensayos. En el ensayo de toxicidad/patogenicidad aguda oral se obtuvo que el microorganismo fue eliminado rápidamente sin provocar infección significativa. Se concluyó que el Bactivec no es patogénico por las vías oral y dérmica a las dosis evaluadas. (AU)

Ausencia de toxicidad / patogenicidad de una formulación de Bacillus sphaericus 2362 (Griselesf) / Lack of toxicity/pathogenicity of a Bacillus sphaericus 2362 formulation (Griselesf)

Los insecticidas microbianos ofrecen alternativas efectivas para el control de muchas plagas de insectos, siendo uno de sus mayores atractivos su especificidad. El Griselesf es un biolarvicida producido en Cuba, que contiene Bacillus sphaericus cepa 2362 como ingrediente activo. Con el objetivo de evaluar su toxicidad/patogenicidad, se administró el Griselesf en una dosis única de 5.6 x 108 unidades formadoras de colonia por vía oral a ratas, y por vía dérmica a dos grupos de conejos albinos, uno recibiendo una dosis de Griselesf de 1.12 x 109 unidades formadoras de colonia por animal, y el otro una dosis de 8.4 x 108 unidades formadoras de colonia de Bacillus sphaericus cepa 2362 por animal. En ambos ensayos, las observaciones clínicas fueron diarias, y se evaluó el comportamiento del peso corporal. Además, en el ensayo por vía oral se estimó el aclaramiento mediante recolección de las heces fecales, y se evaluó la infectividad mediante toma de muestras de fluidos y órganos. Al final de los ensayos se realizó la necropsia a todos los animales. No ocurrieron mortalidades, ni evidencias de patogenicidad o toxicidad relacionada con el tratamiento en ninguno de los ensayos. En el ensayo de toxicidad/patogenicidad aguda oral, el microorganismo fue aclarado rápidamente sin provocar infección significativa. Se concluyó que el Griselesf no es patogénico por las vías oral y dérmica. (AU)

Ensayo de toxicidad a dosis repetidas (28 días) por vía oral del extracto acuoso de Morinda citrifolia en ratas Sprague Dawley / Repeated dose oral toxicity assay (28 days) of the aqueous extract of Morinda citrifolia in Sprague Dawley rats

Se evaluó la toxicidad oral a dosis repetida del extracto acuoso de Morinda citrifolia, obtenido a partir de la planta del mismo nombre, y con reportadas propiedades antitumorales. Nuestro objetivo fue determinar los signos de toxicidad manifiestos tras la administración diaria de 1000 mg/kg de peso corporal a ratas Sprague Dawley durante 28 días, para lo cual se establecieron un grupo control y uno tratado. Se realizaron observaciones clínicas diarias, determinaciones semanales del peso corporal, del consumo de agua y alimento, realizándose al final del estudio la necropsia completa de todos los animales, así como el procesamiento histológico de diferentes órganos, y la determinación de los parámetros hematológicos y bioquímicos. No se produjeron muertes ni alteraciones de signos clínicos, del peso corporal, ni del consumo de agua y alimentos. Los resultados hematológicos y bioquímicos reflejaron ligeras variaciones entre grupos sin significación biológica, ya que todos los parámetros se encontraron dentro del rango fisiológico normal establecido a partir de los controles. No se encontraron lesiones macroscópicas ni microscópicas (AU)

Lung tumor in a patient with congenital unilateral hypoplasia of the pulmonary artery.

The occurrence of an undifferentiated carcinoma in the affected lung of a cigarette smoker with asymptomatic congenital hypoplasia of the right pulmonary artery is reported.