Phenolic acid protects of renal damage induced by ochratoxin A in a 28-days-oral treatment in rats.

The present study aimed to characterize the chlorogenic acid (ChlA) capacity to reverse the toxic effects induced by ochratoxin A (OTA) in a subacute toxicity test in rats. Male Wistar rats were fed orally by gavage for 28 days with OTA (0.4mg/kg bw/day), ChlA (5mg/kg bw/day) or the combination OTA (0.4mg/kg bw/day)+ChlA (5mg/kg bw/day). No deaths, no decrease in feed intake or body weight in any experimental group were recorded. The negative control group and the animals treated with ChlA alone showed no changes in any parameters evaluated. In OTA-treated group significant changes such as decrease in urine volume, proteinuria, occult blood, increase in serum creatinine values; decrease in absolute and relative kidney weight and characteristics histopathological lesions that indicated kidney damage were observed. However, limited effect on oxidative stress parameters were detected in kidneys of OTA-treated group. Animals treated with the combination OTA+ChlA were showed as negative control group in the evaluation of several parameters of toxicity. In conclusion, ChlA, at given concentration, improved biochemical parameters altered in urine and serum and pathological damages in kidneys induced by OTA exposure, showing a good protective activity, but not by an apparent antioxidant mechanism.

In Vitro and In Vivo Cytogenotoxic Effects of Hot Aqueous Extract of Achyrocline satureioides (Lam.) DC.

In this work we extend the toxicological studies of hot aqueous extract of A. satureioides (As-HAE) evaluating cytotoxic and apoptotic effects on human peripheral blood mononuclear cells (PBMCs). We also determine genotoxic action of this extract in vivo. In addition, the extract was chemically characterized. Finally, we established a comparison with previous data of cold aqueous extract. The As-HAE induced cytotoxicity on PBMCs determined by trypan blue dye exclusion (CC50 = 653 µg/mL) and MTT (CC50 = 588 µg/mL) assays being more toxic than cold extract. However, As-HAE as well as cold extract did not induce apoptosis measured by Hoechst 33258 staining, TUNEL assay, and DNA fragmentation analysis. The in vivo micronucleus test showed that As-HAE exerted cytogenotoxic effects on bone marrow of mice, contrary to what was observed with cold extract. The chemical study of As-HAE allowed identifying the flavonoids found in cold extract: luteolin, quercetin, and 3-O-methylquercetin, but at higher concentrations. We suggest that toxic effects induced by As-HAE could be due to high concentrations of these flavonoids. Given that As-HAE is the most used in folkloric medicine, its administration should be controlled in order to prevent potential cell damage.

Polyphenols as possible bioprotectors against cytotoxicity and DNA damage induced by ochratoxin A.

The present study aimed to investigate the protective effects of luteolin (L), chlorogenic acid (ChlA) and caffeic acid (CafA) against cyto-genotoxic effects caused by OTA. Vero cells and rat lymphocytes were used and viability was measured by neutral red uptake, MTT and trypan blue dye exclusion method. L (50 and 100µg/mL), ChlA (100 and 200µg/mL) and CafA (10-50µg/mL) reduced the damage induced by OTA (10µg/mL) on both cells type shown a good protective effect. The comet and micronucleus tests in Balb/c mice were performed. ChlA (10mg/kg bw) reduced OTA (0.85mg/kg bw)-induced DNA damage on blood and bone marrow cells, CafA (10mg/kg bw) showed protective effect only in blood cells and luteolin (2.5mg/kg bw) failed to protect DNA integrity on cells. In conclusion, polyphenols tested reduced the toxicity caused by OTA on different target cells with good protective effect, being ChlA the compound that showed the best effects.

Evaluation of the cytotoxicity, genotoxicity and apoptotic induction of an aqueous extract of Achyrocline satureioides (Lam.) DC.

Achyrocline satureioides is widely consumed as infusion or aperitif and shows important therapeutic properties. Previously, we reported absence of genotoxicity of cold aqueous extract (CAE) of A. satureioides by Allium test. However, one test cannot predict the genotoxic effects of a substance. Thus, the aim of this work was to investigate cytotoxicity, genotoxicity and apoptotic ability of CAE of A. satureioides. In addition, CAE was chemically characterized. The cytotoxicity was evaluated by Trypan blue and MTT assays. The apoptotic capacity was evaluated by Hoechst staining and DNA fragmentation-analysis. The genotoxicity was studied by comet assay (CA) and micronucleus test. The identification and quantification of flavonoids were performed by HPLC-ESI-MS/MS. The cytotoxicity studies indicated low toxicity of CAE. In addition, CAE did not induce apoptotic effects on human PBMCs. CAE did not show genotoxicity in vitro against Vero cells, at 10-50 µg/mL. CAE did not induce in vivo genotoxic effects, but it showed at high concentrations cytotoxicity by micronucleus assay. CAE presented flavonoids such as quercetin, 3-O-methylquercetin and luteolin. In conclusion, A. satureioides at popularly concentrations used, in aperitif or infusion, can be consumed safely because did not show any cytotoxic or genotoxic effects.

Propiedades inmunológicas y mitogénicas de productos derivados de Minthostachys verticillata sobre linfocitos de niños alérgicos con infección viral / Immunological and mitogenis properties of Minthostachys verticillata derivates products on lymphocytes from allergic children with viral infection

Antecedentes. La decocción y el aceite esencial de Minthostachys verticillata han demostrado efectos antimicrobianos sobre el género Staphylococcus y propiedad antiherpéticas. Objetivos. Determinar la capacidad inmunogénica de la decocción vegetal en inmunizaciones experimentales. Investigar la capacidad proliferativa de linfocitos estimulados con derivados de M. verticillata. Evaluar los efectos de la dococción sobre el mitógeno pokeweed (PWM). Detectar y cuantificar LT CD8+ y LB en células proliferadas provenientes de niños alérgicos con infección VHS-1. Materiales y métodos. Se inmunizaron 2 conejos con la decocción. Se evaluaron los anticuerpos por precipitación y hemaglutinación pasiva. Se estudió la proliferación de linfocitos de 25 pacientes de 2 a 14 años, 15 alérgicos con lesiones de Herpes Simplex Tipo 1 (VHS-1) y 10 controles. Las células fueron estimuladas con a) decocción, b) aceite esencial, c) PWM, d) decocción + PWM, e) control. Se caracterizaron y cuantificaron LT CD8+ y LB por IFD. Resultados. La decocción indujo la síntesis de anticuerpos no precipitantes y hemaglutinantes de glóbulos rojos de carnero. Los linfocitos de los pacientes con VHS-1 mostraron niveles linfoproliferativos más elevados(p<0,01). La decocción y el aceite mostraron efectos mitogénicos similares a PWM. El agregado simultáneo de decocción y PWM redujo la proliferación alcamzada por cada uno (p<0,001) tanto en infectados como en controles. Entre células proliferadas se caracterizaron LT CD8+ y LB. Conclusión. La decocción fue inmunogénica, los anticuerpos tuvieron características de coprecipitantes. Los derivados de M. verticillata mostraron propiedades mitogénicas similares a PWM. La decocción inhibió los efectos de PWM

Propiedades inmunológicas y mitogénicas de productos derivados de Minthostachys verticillata sobre linfocitos de niños alérgicos con infección viral / Immunological and mitogenis properties of Minthostachys verticillata derivates products on lymphocytes from allergic children with viral infection

Antecedentes. La decocción y el aceite esencial de Minthostachys verticillata han demostrado efectos antimicrobianos sobre el género Staphylococcus y propiedad antiherpéticas. Objetivos. Determinar la capacidad inmunogénica de la decocción vegetal en inmunizaciones experimentales. Investigar la capacidad proliferativa de linfocitos estimulados con derivados de M. verticillata. Evaluar los efectos de la dococción sobre el mitógeno pokeweed (PWM). Detectar y cuantificar LT CD8+ y LB en células proliferadas provenientes de niños alérgicos con infección VHS-1. Materiales y métodos. Se inmunizaron 2 conejos con la decocción. Se evaluaron los anticuerpos por precipitación y hemaglutinación pasiva. Se estudió la proliferación de linfocitos de 25 pacientes de 2 a 14 años, 15 alérgicos con lesiones de Herpes Simplex Tipo 1 (VHS-1) y 10 controles. Las células fueron estimuladas con a) decocción, b) aceite esencial, c) PWM, d) decocción + PWM, e) control. Se caracterizaron y cuantificaron LT CD8+ y LB por IFD. Resultados. La decocción indujo la síntesis de anticuerpos no precipitantes y hemaglutinantes de glóbulos rojos de carnero. Los linfocitos de los pacientes con VHS-1 mostraron niveles linfoproliferativos más elevados(p<0,01). La decocción y el aceite mostraron efectos mitogénicos similares a PWM. El agregado simultáneo de decocción y PWM redujo la proliferación alcamzada por cada uno (p<0,001) tanto en infectados como en controles. Entre células proliferadas se caracterizaron LT CD8+ y LB. Conclusión. La decocción fue inmunogénica, los anticuerpos tuvieron características de coprecipitantes. Los derivados de M. verticillata mostraron propiedades mitogénicas similares a PWM. La decocción inhibió los efectos de PWM (AU)

Potency analysis of inactivated vaccines for Aujeszky's disease virus (ADV), strain RC/79: lymphocyte stimulation in immunized pigs.

Mitogenic and antigenic lymphocyte stimulation was examined in pigs that had been immunized with 2 inactivated vaccines which had been prepared with Aujesky's disease virus (ADV), strain RC/79. One vaccine was partially purified by ultra-centrifugation (Vaccine A) the other one was not (Vaccine B). A second dose of vaccine had no influence over the blastogenic response when the lymphocytes were stimulated with phytohemoagglutinin (PHA). Lymphocyte response to the ADV antigen in the immunized pigs was significantly higher at day 30 post inoculation than at day 0 indicating that it was highly specific. Cellular antigens contained in the viral cultures produced a slight non-specific response as shown by a low increase in the levels of lymphocyte blastic transformation (LBT) in the control group at day 30 p.i., this group only received a non infected Vero cell suspension. This was the case in pigs that received vaccine A as well as in those that were vaccinated with vaccine B. Vaccine B contains a greater quantity of contaminating cellular antigens, since it is an impure vaccine. Such antigens could act as non-specific immunomodulators, potentiating cell-mediated immunity (CMI). This assay demonstrated that inactivated vaccines produced with VPR-RC/79, partially purified and unpurified are capable of inducing a humoral immune response. The blastogenic reaction of the peripheral blood lymphocytes to antigens of ADV strain RC/79, indicated that the employed immunogens also induced the CMI. Results indicate that the analyzed immunogens could be considered for the possible implementation of epidemiological measures, which imply the use of vaccines to prevent pseudo-rabies in Argentina.

[Herpes simplex: generalized infection in a newborn]. / Virus herpes simple: infección generalizada en un neonato.

BACKGROUND: Most mucose-cutaneous infections can be diagnosed by clinical history with explorative technique. Nevertheless the definitive etiologic diagnostic can only be established with the help of the isolation and identification of the causal agent. PATIENT AND METHODS: We reported a case of generalized infection in a new born, with is clinical characteristics, virological diagnostic techniques and treatment of the disease. RESULTS: Herpes simplex virus was isolated in Vero cellular culture from CRL and vesiculose lesions of the child's thorax region and from the mothers endocervical scrapes. The agent was identified by seroneutralization test. This confirmed a perinatal transmission of Herpes simplex virus type 2. The tomographic studies revealed characteristic alterations of a viral encephalitis. An antiviral treatment was useful in this therapy. CONCLUSIONS: Our results demonstrate the importance of an early viral diagnostic which permits the applications of specific treatment and thus the prevention of severe complications.

[Experimental infection of pregnant gilts with Aujeszky's disease virus strain RC/79]. / Infección experimental de cerdas preñadas con virus de la enfermedad de Aujeszky cepa RC/79.

The RC/79 strain of the Aujeszky's disease virus was able to induce reproductive failure of pregnant gilts intranasally inoculated at different gestation periods. Four gilts 40-46 days pregnant (group A) and 6 gilts 70-73 days pregnant (group B) were instilled with 0.2 ml x 10(5) tissue culture infectious dose 50 (TCID50/0.2 ml) of the RC/79 strain into each nostril. Two gilts 70-73 days pregnant (group C) were used as non exposed controls. The three groups were kept in separated boxes and they were observed for clinical signs of infections and samples were collected for determination of viral shedding every day. Viral isolation was attempted in Vero cells (figure 1). From the 2nd to 7th day after inoculation, groups A and B showed fever anorexia, sneezing, coughing and depression; and viral isolation from nasal swabs was possible in 7 gilts at days 4 to 11, 9 gilts developed neutralizing antibodies. The virus caused fetal reabsorption in swine during the first period of pregnancy (group A), while infection during late pregnancy resulted in still birth or normal pigs and one mummification (group B). The entire a live litter was composed of no more than 8 suckling pigs in both groups. At necropsy virus from turbinates, ovary , placenta, spleen and lung could be isolated only from 3 gilts (group B, table 1). In 5 of 35 stillbirth and alive fetuses virus could be isolated from spleen (100%), lung (80%), liver (60%) and brain (40%) indicating that the virus has the ability to cross the placental barrier thus producing lesions in porcine fetuses and causing reproductive failure in sows (table 2). Tissue specimens from these 35 fetuses were fixed in 10% formalin, included in paraffin sectioned and stained with hematoxylin and eosin. In 13 fetuses microscopic lesions i.e. necrotic foci were found in lung (60%), liver (40%) and spleen (20%), these alterations were coincident with gross lesions in most of them. Inclusion bodies were absent. The gilts organs did not present gross lesions.

Infección experimental de cerdas preñadas con virus de la enfermedad de Aujeszky cepa RC/79 / [Experimental infection of pregnant gilts with Aujeszky’s disease virus strain RC/79]

The RC/79 strain of the Aujeszky’s disease virus was able to induce reproductive failure of pregnant gilts intranasally inoculated at different gestation periods. Four gilts 40-46 days pregnant (group A) and 6 gilts 70-73 days pregnant (group B) were instilled with 0.2 ml x 10(5) tissue culture infectious dose 50 (TCID50/0.2 ml) of the RC/79 strain into each nostril. Two gilts 70-73 days pregnant (group C) were used as non exposed controls. The three groups were kept in separated boxes and they were observed for clinical signs of infections and samples were collected for determination of viral shedding every day. Viral isolation was attempted in Vero cells (figure 1). From the 2nd to 7th day after inoculation, groups A and B showed fever anorexia, sneezing, coughing and depression; and viral isolation from nasal swabs was possible in 7 gilts at days 4 to 11, 9 gilts developed neutralizing antibodies. The virus caused fetal reabsorption in swine during the first period of pregnancy (group A), while infection during late pregnancy resulted in still birth or normal pigs and one mummification (group B). The entire a live litter was composed of no more than 8 suckling pigs in both groups. At necropsy virus from turbinates, ovary , placenta, spleen and lung could be isolated only from 3 gilts (group B, table 1). In 5 of 35 stillbirth and alive fetuses virus could be isolated from spleen (100

) and brain (40

) indicating that the virus has the ability to cross the placental barrier thus producing lesions in porcine fetuses and causing reproductive failure in sows (table 2). Tissue specimens from these 35 fetuses were fixed in 10

formalin, included in paraffin sectioned and stained with hematoxylin and eosin. In 13 fetuses microscopic lesions i.e. necrotic foci were found in lung (60

) and spleen (20

), these alterations were coincident with gross lesions in most of them. Inclusion bodies were absent. The gilts organs did not present gross lesions.

Infección experimental de cerdas preñadas con virus de la enfermedad de Aujeszky cepa RC/79 / [Experimental infection of pregnant gilts with Aujeszky’s disease virus strain RC/79]

The RC/79 strain of the Aujeszky’s disease virus was able to induce reproductive failure of pregnant gilts intranasally inoculated at different gestation periods. Four gilts 40-46 days pregnant (group A) and 6 gilts 70-73 days pregnant (group B) were instilled with 0.2 ml x 10(5) tissue culture infectious dose 50 (TCID50/0.2 ml) of the RC/79 strain into each nostril. Two gilts 70-73 days pregnant (group C) were used as non exposed controls. The three groups were kept in separated boxes and they were observed for clinical signs of infections and samples were collected for determination of viral shedding every day. Viral isolation was attempted in Vero cells (figure 1). From the 2nd to 7th day after inoculation, groups A and B showed fever anorexia, sneezing, coughing and depression; and viral isolation from nasal swabs was possible in 7 gilts at days 4 to 11, 9 gilts developed neutralizing antibodies. The virus caused fetal reabsorption in swine during the first period of pregnancy (group A), while infection during late pregnancy resulted in still birth or normal pigs and one mummification (group B). The entire a live litter was composed of no more than 8 suckling pigs in both groups. At necropsy virus from turbinates, ovary , placenta, spleen and lung could be isolated only from 3 gilts (group B, table 1). In 5 of 35 stillbirth and alive fetuses virus could be isolated from spleen (100

) and brain (40

) indicating that the virus has the ability to cross the placental barrier thus producing lesions in porcine fetuses and causing reproductive failure in sows (table 2). Tissue specimens from these 35 fetuses were fixed in 10

formalin, included in paraffin sectioned and stained with hematoxylin and eosin. In 13 fetuses microscopic lesions i.e. necrotic foci were found in lung (60

) and spleen (20

), these alterations were coincident with gross lesions in most of them. Inclusion bodies were absent. The gilts organs did not present gross lesions.

Infección experimental de cerdas preñadas con virus de la enfermedad de Aujeszky cepa RC/79. / [Experimental infection of pregnant gilts with Aujeszkys disease virus strain RC/79]

The RC/79 strain of the Aujeszkys disease virus was able to induce reproductive failure of pregnant gilts intranasally inoculated at different gestation periods. Four gilts 40-46 days pregnant (group A) and 6 gilts 70-73 days pregnant (group B) were instilled with 0.2 ml x 10(5) tissue culture infectious dose 50 (TCID50/0.2 ml) of the RC/79 strain into each nostril. Two gilts 70-73 days pregnant (group C) were used as non exposed controls. The three groups were kept in separated boxes and they were observed for clinical signs of infections and samples were collected for determination of viral shedding every day. Viral isolation was attempted in Vero cells (figure 1). From the 2nd to 7th day after inoculation, groups A and B showed fever anorexia, sneezing, coughing and depression; and viral isolation from nasal swabs was possible in 7 gilts at days 4 to 11, 9 gilts developed neutralizing antibodies. The virus caused fetal reabsorption in swine during the first period of pregnancy (group A), while infection during late pregnancy resulted in still birth or normal pigs and one mummification (group B). The entire a live litter was composed of no more than 8 suckling pigs in both groups. At necropsy virus from turbinates, ovary , placenta, spleen and lung could be isolated only from 3 gilts (group B, table 1). In 5 of 35 stillbirth and alive fetuses virus could be isolated from spleen (100

), lung (80

), liver (60

) and brain (40

) indicating that the virus has the ability to cross the placental barrier thus producing lesions in porcine fetuses and causing reproductive failure in sows (table 2). Tissue specimens from these 35 fetuses were fixed in 10

formalin, included in paraffin sectioned and stained with hematoxylin and eosin. In 13 fetuses microscopic lesions i.e. necrotic foci were found in lung (60

), liver (40

) and spleen (20

), these alterations were coincident with gross lesions in most of them. Inclusion bodies were absent. The gilts organs did not present gross lesions.

Infección experimental de cerdas preñadas con virus de la enfermedad de Aujeszky cepa RC/79. / [Experimental infection of pregnant gilts with Aujeszkys disease virus strain RC/79]

The RC/79 strain of the Aujeszkys disease virus was able to induce reproductive failure of pregnant gilts intranasally inoculated at different gestation periods. Four gilts 40-46 days pregnant (group A) and 6 gilts 70-73 days pregnant (group B) were instilled with 0.2 ml x 10(5) tissue culture infectious dose 50 (TCID50/0.2 ml) of the RC/79 strain into each nostril. Two gilts 70-73 days pregnant (group C) were used as non exposed controls. The three groups were kept in separated boxes and they were observed for clinical signs of infections and samples were collected for determination of viral shedding every day. Viral isolation was attempted in Vero cells (figure 1). From the 2nd to 7th day after inoculation, groups A and B showed fever anorexia, sneezing, coughing and depression; and viral isolation from nasal swabs was possible in 7 gilts at days 4 to 11, 9 gilts developed neutralizing antibodies. The virus caused fetal reabsorption in swine during the first period of pregnancy (group A), while infection during late pregnancy resulted in still birth or normal pigs and one mummification (group B). The entire a live litter was composed of no more than 8 suckling pigs in both groups. At necropsy virus from turbinates, ovary , placenta, spleen and lung could be isolated only from 3 gilts (group B, table 1). In 5 of 35 stillbirth and alive fetuses virus could be isolated from spleen (100

), lung (80

), liver (60

) and brain (40

) indicating that the virus has the ability to cross the placental barrier thus producing lesions in porcine fetuses and causing reproductive failure in sows (table 2). Tissue specimens from these 35 fetuses were fixed in 10

formalin, included in paraffin sectioned and stained with hematoxylin and eosin. In 13 fetuses microscopic lesions i.e. necrotic foci were found in lung (60

), liver (40

) and spleen (20

), these alterations were coincident with gross lesions in most of them. Inclusion bodies were absent. The gilts organs did not present gross lesions.