Effect of Lactiplantibacillus plantarum cell-free culture on bacterial pathogens isolated from cystic fibrosis patients: in vitro and in vivo studies.

This study aimed to investigate the effects of the cell-free supernatant of Lactiplantibacillus plantarum ATCC® 10241TM on the biofilm-forming capacity of Pseudomonas aeruginosa strains isolated from cystic fibrosis (CF) patients. In addition, the study evaluated the in vivo potential of the cell-free supernatant to modulate inflammation and reduce lung damage in mice infected with P. aeruginosa strains or co-challenged with P. aeruginosa and the Streptococcus milleri group (SMG). The results showed that CF-derived P. aeruginosa strains can infect the respiratory tract of adult mice, inducing local inflammation and lung damage. The severity of these infections was exacerbated when P. aeruginosa was co-administered with SMG. Notably, nebulization with the cell-free supernatant of L. plantarum produced beneficial effects, reducing respiratory infection severity and inflammatory responses induced by P. aeruginosa, both alone or in combination with SMG. Reduced bacterial loads and lung damage were observed in supernatant-treated mice compared to controls. Although further mechanistic studies are necessary, the results show that the cell-free supernatant of L. plantarum ATCC® 10241TM is an interesting adjuvant alternative to treat P. aeruginosa respiratory infections and superinfections in CF patients.

Novel topical formulation for ischemic chronic wounds. Technological design, quality control and safety evaluation.

Ulceration of the foot in diabetes is common and disabling, and frequently leads to amputation of the leg. The pathogenesis of foot ulceration is complex, clinical presentation variable and management requires early expert assessment. Despite treatment, ulcers readily become chronic wounds. Chronic wounds are those that remain in a chronic inflammatory state failing a normal healing process patterns. This is partially caused by inefficient eradication of opportunistic pathogens like Pseudomonas aeruginosa. We propose its control or eradication will promote wound healing. Lactobacillus plantarum cultures supernatants (LAPS) shows antipathogenic and pro-healing properties. The main objective was to design two pharmaceutical dosage forms by using LAPS as active pharmaceutical ingredient and to perform its quality control, in vitro activity conservation tests and human trials (safety evaluation). Both selected formulations reach the technological quality expected for 120 days, shows adequate occlusive characteristics and proper adhesion to human skin. From the in vitro release assays were found that LAPS shows adequate release from matrix and maintain its antimicrobial and anti-biofilm activity. First human trials were developed and neither edema nor erythema on healthy skin voluntaries was found. We conclude that C80 and C100 are adequate for their use in future clinical trials to demonstrate a comprehensive therapeutic effectiveness in ischemic chronic wounds.

Equipment and method for in vitro release measurements on topical dosage forms.

CONTEXT: In countries where research budgets are meager as Argentina, the tendency to innovation and improvements in the designs prototypes "made in Argentina" marks a growing trend adopted by researchers. This article presents a diffusion cell of original design, for release studies of Active Pharmaceutical Ingredient (API) from classical topical dosage forms, also includes the methodology for its optimization and validation. The objective was to evaluate and validate a system designed and to compare it to the Franz cells system. METHODS: Parameters, reproducibility and robustness were performed included factors as, stirring conditions, membrane stabilization treatment and temperature variation. Release and retention on membrane assay were performed using two different API and formulations. RESULTS: The method is reproducible and robust for the parameters tested. Release assays show that no significative difference with the Franz Cells system. Our system allows the simultaneous measurement of different parameters, representing an innovation on these methodologies. The LMC was used for assays of in vitro retention on membrane and the values obtained were reproducible and coincident whit values obtained for other authors. CONCLUSIONS: The system designed and the methodology employed, are acceptable for in vitro release studies. The device and method has the characteristics required.

Formulation and quality control of semi-solid containing harmless bacteria by-products: chronic wounds pro-healing activity.

Chronic wounds are those that remain in a chronic inflammatory state and fail to follow normal healing process. Infection is one of the most important causes of chronicity. A frequent pathogen isolated from chronic infections is Pseudomonas aeruginosa; refractory to therapy and host immune attack in its biofilm phenotype. Lactobacillus plantarum cultures supernatants (LAPS) interfere with its pathogenic capacity. In addition, LAPS showed bacteriostatic and bactericide properties and is neither cytotoxic nor an inductor of necrosis-apoptosis. LAPSs chemical composition was determined; allowing us to propose a correlation between its constituents and their biological activity. This article shows a pharmaceutical dosage form designed by using LAPS as an API with pro-healing activity and its quality control. Pharmacotechnical and anti-microbial assays were adapted to demonstrate that the vehicle used does not modify LAPS activities. Selected formulation (F100) showed fair mechanical and technological properties. From the in vitro release assays was found an adequate release from the carrier matrix and maintains its anti-pathogenic activity for 6 months. We propose F100 for chronic wounds treatment. The use of harmless bacteria by-products, such as LAPS, to antagonize infectious pathogens that have ability to form biofilm is an efficient and economic approach to treat infected chronic wounds.

HPV knowledge in Mexican college students: implications for intervention programmes.

In order to promote new human papillomavirus (HPV) prevention and detection methods effectively in Mexico, it is important to understand how much the population knows about the virus. This study aimed to determine the demographic and behavioural factors associated with HPV awareness and knowledge in a population of Mexican college students. With a response rate of 77%, data were collected from 1109 college students aged 17-25 years old at the Autonomous University of the State of Morelos in 2006. Students completed a questionnaire that assessed demographic and behavioural characteristics along with questions about HPV. A small percentage (16.9%) of the college students had never heard about HPV. Characteristics associated with not having heard about HPV included being male, not having running water, not having health insurance and not having sexual experience. Students had a median score of 5 out of 10 on an HPV knowledge index based on 10 yes/no questions about HPV developed for this study. Students had higher HPV knowledge scores if they studied health science, or science and engineering, were a fourth year student, had running water at home, had health insurance, or were a female who had had a previous Pap smear. Although most of these Mexican college students had heard of HPV, they had limited knowledge about the virus and prevention strategies. Further research in Mexican college students is needed to explain the variations in HPV knowledge to create appropriate health education programmes.

Effects of plant lactones on the production of biofilm of Pseudomonas aeruginosa.

Sixteen plant sesquiterpene lactones, thirteen from four species of the Family Asteraceae, and three from a species of Hepaticae, as well as seven annonaceous acetogenins isolated from the seeds of the tropical tree Annona cherimolia (Family Annonaceae), were evaluated for their ability to inhibit or stimulate the production of biofilm by a strain of Pseudomonas aeruginosa. The tested compounds carry a gamma-lactone moiety in their structures. This structural feature is similar to the lactone moiety present in N-acyl homoserine lactones, compounds that play the important role of "quorum sensors" in the mechanisms of biofilm formation observed in many gram-negative bacteria. A special assay was employed to evaluate the influence of the tested plant compounds to inhibit or stimulate the production of biofilm in a P. aeruginosa wild strain. Most of the tested compounds affected the biofilm formation mechanism. Six sesquiterpene lactones isolated from Acanthospermum hispidum and one from Enydra anagallis as well as an acetogenin from Annona cherimolia strongly inhibited (69-77%) the biofilm formation when incorporated to a bacterial culture at a concentration of 2.5 microg/ml. However, one of the acetogenins, squamocin, stimulated the biofilm formation even at a concentration of 0.25 microg/ml. The study of substances affecting the biofilm formation can lead to the design of new strategies to control P. aeruginosa infections.

Efecto de la carragenina e indometacina sobre el crecimiento de un fibrosarcoma murino / Effect of carrageena and indomethacin on the growth of a murine fibrosarcoma

Los tumores sólidos para crecer más de 2 mm, necesitan desarrollar nuevos vasos sanguíneos. Las células neoplásicas secretan factores de crecimiento que estimulan la angiogénesis y el crecimiento tumoral. La Carragenina bloquea un vitro la unión de los factores de crecimiento a sus receptores. Ensayamos in vivo su acción con el objetivo de analizar si inhibe el desarrollo de un fibrosarcoma murino. Para neutralizar la acción inflamatoria de la Carragenina usamos la Indometacina, que es un antiinflamatorio no esteroide. El tumor usado fue en fibrosarcoma inducido con metilcolanterno en ratones Balb/c y mantenido por pasaje seriado de células tumorales, en ratones de la misma cepa. El volumen de los tumores fue evaluado midiendo dos dimensiones y aplicando la fórmula V = 0.4 x d(2) x D. Los ratones con tumores fueron separados en grupos, uno de los cuales se usó como testigo y los otros tratados en forma separada con indometacina, Carragenina y Carragenina-Indometacina. Se comparó el volumen de los tumores de los ratones tratados con respecto al testigo y el de los tratados entre sí, utilizando el Test t de Student. Se demostró que la Carregenina y la indometacina, inhiben el desarrollo del fibrosarcoma. La acción inhibitoria de la Carragenina sobre el crecimiento tumoral es significativamente mayor que el efecto antitumoral de la Indometacina y el de la Corragenina-Indometacina. (AU)

Efecto de la carragenina e indometacina sobre el crecimiento de un fibrosarcoma murino / Effect of carrageena and indomethacin on the growth of a murine fibrosarcoma

Los tumores sólidos para crecer más de 2 mm, necesitan desarrollar nuevos vasos sanguíneos. Las células neoplásicas secretan factores de crecimiento que estimulan la angiogénesis y el crecimiento tumoral. La Carragenina bloquea un vitro la unión de los factores de crecimiento a sus receptores. Ensayamos in vivo su acción con el objetivo de analizar si inhibe el desarrollo de un fibrosarcoma murino. Para neutralizar la acción inflamatoria de la Carragenina usamos la Indometacina, que es un antiinflamatorio no esteroide. El tumor usado fue en fibrosarcoma inducido con metilcolanterno en ratones Balb/c y mantenido por pasaje seriado de células tumorales, en ratones de la misma cepa. El volumen de los tumores fue evaluado midiendo dos dimensiones y aplicando la fórmula V = 0.4 x d(2) x D. Los ratones con tumores fueron separados en grupos, uno de los cuales se usó como testigo y los otros tratados en forma separada con indometacina, Carragenina y Carragenina-Indometacina. Se comparó el volumen de los tumores de los ratones tratados con respecto al testigo y el de los tratados entre sí, utilizando el Test t de Student. Se demostró que la Carregenina y la indometacina, inhiben el desarrollo del fibrosarcoma. La acción inhibitoria de la Carragenina sobre el crecimiento tumoral es significativamente mayor que el efecto antitumoral de la Indometacina y el de la Corragenina-Indometacina.