Action of methicillin on the "in vitro" growth of bacteria Staphyloccocus aureus methicillin-resistance previously treated with homeopathic dilutions

Background: Methicillin-resistant Staphylococcus aureus (MRSA) causes nosocomial infections, and it has been considered as a worldwide epidemic. The medical system seeks new strategies to fight against MRSA that do not generate resistant strains to antibiotics. Homeopathy has been explored as one of these new strategies, which may play a pivotal role. In this context, we conducted studies on the action of homeopathy on growth of MRSA bacteria in vitro. The results showed a decrease in growth of bacterial strains with homeopathic dilutions of Belladonna and the S. aureus nosode. Now we have proposed to evaluate the minimum inhibitory concentration (MIC) of the antibiotic methicillin or oxacillin on S. aureus MRSA, previously incubated with the homeopathic dilutions of Belladonna or S. aureus nosode.&#x0D; Methods: The Clinical and Laboratory Standards Institute (CLSI 2014) standards were followed according to the determination of the minimum inhibitory concentration (MIC). In 5 mL of cation adjusted Mueller Hinton (CAMH) broth, it was added 420 µl of 30% alcohol or Belladonna and S. aureus’ nosode in the dilutions 6cH, 12cH and 30cH. Then a 20µl of bacterial suspension of MRSA was added to 0.5 McFarland range and diluted to 1/10. The tubes were incubated in an oven at 37⁰C for three hours. The plates were previously prepared with 50µl per well of serial dilutions of the antibiotic oxacillin in concentrations of 128 µg/mL to 0.5 µg/mL in CAMH broth. Then it was added 50 µl per well of bacterial cultures. The plate was incubated in an oven at 37⁰C for 24 hours and the bacterial growth measured in a spectrophotometer 600nm. The point of the MIC of oxacillin for S. aureus is 4 µg/mL, according to CLSI 2014 criteria.&#x0D; Results: We did not observe the total inhibition of bacterial growth when incubated with the homeopathic medicine and oxacillin. In evaluation of the spectrophotometer culture, we observed significant changes in the growth, compared to the control (30% alcohol). Cultures treated with Belladonna 6cH and the antibiotic in the dilution 4 µg/mL showed a decrease of 40% of the growth, while in the 30cH the drop was of 75%. Cultures treated with the S. aureus nosode 30cH and the antibiotic at 4 µg/mL dilution, showed a decrease of 60% in bacterial growth in vitro.&#x0D; Conclusion: The results suggest that bacterial cultures the S. aureus (MRSA) incubated with the homeopathic medicines would be more susceptible to oxacillin’s antimicrobial action.</jats:p

Identification of calgranulin B interacting proteins and network analysis in gastrointestinal cancer cells

Calgranulin B is known to be involved in tumor development, but the underlying molecular mechanism is not clear. To gain insight into possible roles of calgranulin B, we screened for calgranulin B-interacting molecules in the SNU-484 gastric cancer and the SNU-81 colon cancer cells. Calgranulin B-interacting partners were identified by yeast two-hybrid and functional information was obtained by computational analysis. Most of the calgranulin B-interacting partners were involved in metabolic and cellular processes, and found to have molecular function of binding and catalytic activities. Interestingly, 46 molecules in the network of the calgranulin B-interacting proteins are known to be associated with cancer and FKBP2 was found to interact with calgranulin B in both SNU-484 and SNU-81 cells. Polyubiquitin-C encoded by UBC, which exhibited an interaction with calgranulin B, has been associated with various molecules of the extracellular space and plasma membrane identified in our screening, including Na-K-Cl cotransporter 1 and dystonin in SNU-484 cells, and ATPase subunit beta-1 in SNU-81 cells. Our data provide novel insight into the roles of calgranulin B of gastrointestinal cancer cells, and offer new clues suggesting calgranulin B acts as an effector molecule through which the cell can communicate with the tumor microenvironment via polyubiquitin-C

Expression of Cellular Components in Granulomatous Inflammatory Response in Piaractus mesopotamicus Model

The present study aimed to describe and characterize the cellular components during the evolution of chronic granulomatous inflammation in the teleost fish pacus (P. mesopotamicus) induced by Bacillus Calmette-Guerin (BCG), using S-100, iNOS and cytokeratin antibodies. 50 fish (120 +/- 5.0 g) were anesthetized and 45 inoculated with 20 mu L (40 mg/mL) (2.0 x 10(6) CFU/mg) and five inoculated with saline (0,65%) into muscle tissue in the laterodorsal region. To evaluate the inflammatory process, nine fish inoculated with BCG and one control were sampled in five periods: 3rd, 7th, 14th, 21st and 33rd days post-inoculation (DPI). Immunohistochemical examination showed that the marking with anti-S-100 protein and anti-iNOS antibodies was weak, with a diffuse pattern, between the third and seventh DPI. From the 14th to the 33rd day, the marking became stronger and marked the cytoplasm of the macrophages. Positivity for cytokeratin was initially observed in the 14th DPI, and the stronger immunostaining in the 33rd day, period in which the epithelioid cells were more evident and the granuloma was fully formed. Also after the 14th day, a certain degree of cellular organization was observed, due to the arrangement of the macrophages around the inoculated material, with little evidence of edema. The arrangement of the macrophages around the inoculum, the fibroblasts, the lymphocytes and, in most cases, the presence of melanomacrophages formed the granuloma and kept the inoculum isolated in the 33rd DPI. The present study suggested that the granulomatous experimental model using teleost fish P. mesopotamicus presented a similar response to those observed in mammals, confirming its importance for studies of chronic inflammatory reaction.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Sao Paulo State Univ, Sch Agrarian &Vet Sci, Dept Vet Pathol, Sao Paulo, BrazilUniv Santiago de Compostela, Coll Vet, Dept Pathol Anat, Lugo, SpainSao Paulo State Univ, Sch Agrarian &Vet Sci, Dept Vet Pathol, Sao Paulo, BrazilFAPESP: 2010/086245FAPESP: 2009/17640-