IN VITRO ACTIVITY OF VARIOUS POTENCIES OF HOMEOPATHIC DRUG THUJA AGAINST MOLDS INVOLVED IN MYCOTIC KERATITIS

Objective: Isolation and characterisation of clinically isolated fungi from mycotic keratitis and exploration of the in vitro efficacy of various potencies of homeopathic preparations of Thuja occidentalis on the ocular fungal isolates. Methods: Clinical samples were collected from fungal keratitis patients attending a tertiary eye care hospital in Coimbatore, Tamilnadu state, India. The scrapings were also subjected to Gram staining and 10% KOH mount to detect the presence of fungal hyphae. The fungal isolates were subjected to lacto phenol cotton blue (LCB) mount employing cello tape flag method. Homeopathic drug Thuja occidentalis with various potencies viz., Q, 30 C, 200 C, 1 M, 10 M and 50 M were investigated for the growth inhibition of various fungal isolates by plate assay method. Further, a follow up analyses with varying dilutions of Q and 10 M homeopathic potencies of Thuja was carried out for the determination of minimum inhibitory concentration of both microdilution and minimum fungicidal concentration for the Biopolaris isolates. Results: Out of 35 samples analysed, Fusarium spp. (n=5), Aspergillus flavus (n=6), Bipolaris spp. (n=3), Exserohilum spp. (n=3) and Curvularia spp (n=3) were identified. All the potencies of Thuja had good inhibitory activity against Bipolaris spp., followed by Curvularia spp., Exserohilum spp. and Aspergillus flavus. Statistical analysis revealed significant inhibition of all the test isolates by Thuja Q and 50 M. Significant growth inhibition was exhibited by Thuja 30, 200 C, I M for Bipolaris, Exserohilum & A. flavus isolate and Thuja 10 M for all the isolates tested except Fusaria. It was revealed that for the Biopolaris isolates BS1, BS2, BS3 Thuja 10 M and Thuja Q had MIC and MFC of 0.125/10[20,00]0& 0.0625/10 and 0.25/10[20][00]0 & 0.125/10, respectively. Conclusion: The present investigation concludes that homeopathic drug Thuja has good inhibitory activity against the fungi causing keratitis, irrespective of the potencies. It is evident that no definite co-relation exists between various potencies of the same homeopathic drug with regard to their antimycotic properties

Identification of novel regulators of the Nuclear Factor Kappa B pathway in human macrophages

The nuclear factor-kappa B (NF-κB) family of transcription factors has a central role in coordinating the expression of genes that control inflammation, immune responses, cell-proliferation, and a variety of other processes. Ever since its discovery in 1986 in David Baltimore’s lab, the NF-κB pathway has been the prime model of inducible transcription in various cell types, and in response to multiple stimuli. Despite being one of the most well-studied pathway in biology, it still has a lot of unanswered questions associated with it including the events that lead up to its activation in the cytoplasm as well as the sequence of events that lead to the transcription of hundreds of its target genes in the nucleus. Given the pathway’s implication in development and diseases, it has become increasingly important to answer these questions. Here, we present two whole-genome RNAi screens conducted to find novel regulators of this pathway in the physiologically relevant human macrophages in response to Lipopolysaccharides (LPS) and Tumor Necrosis Factor Alpha (TNF). After three levels of screening we have found over 25 potential novel regulators of this pathway, summarized in Chapters 2 and 4. The top hit is the splicing factor and transcriptional co-activator SNW1. We have further validated it as a regulator of the NF-κB pathway in response to multiple stimuli and in five different cell lines (THP-1, U87, 293T, A549, and U2-OS). SNW1 does not seem to affect general constitutive transcription in THP-1 cells but does seem to repress some transcription programs e.g. CREB and NFE2. SNW1 does not regulate the cytoplasmic part of the NF-κB pathway but does complex with the NF-κB hetero dimer in the nucleus on pathway activation. We have shown that it binds to NF-κB’s transcriptional elongation partner p-TEFb and helps recruit it to the NF-κB nuclear complex that contains RNA Polymerase II. We have also shown that SNW1 loses binding from its splicing complex (SNRNP200, SNRNP220) on NF-κB activation. SNW1 is a unique protein shown to be involved in both splicing and transcription and in the case of NF-κB, its role seems to involve recruitment of p-TEFb for effective transcriptional elongation of NF-κB target genes

Identification of novel regulators of the Nuclear Factor Kappa B pathway in human macrophages

The nuclear factor-kappa B (NF-κB) family of transcription factors has a central role in coordinating the expression of genes that control inflammation, immune responses, cell-proliferation, and a variety of other processes. Ever since its discovery in 1986 in David Baltimore’s lab, the NF-κB pathway has been the prime model of inducible transcription in various cell types, and in response to multiple stimuli. Despite being one of the most well-studied pathway in biology, it still has a lot of unanswered questions associated with it including the events that lead up to its activation in the cytoplasm as well as the sequence of events that lead to the transcription of hundreds of its target genes in the nucleus. Given the pathway’s implication in development and diseases, it has become increasingly important to answer these questions. Here, we present two whole-genome RNAi screens conducted to find novel regulators of this pathway in the physiologically relevant human macrophages in response to Lipopolysaccharides (LPS) and Tumor Necrosis Factor Alpha (TNF). After three levels of screening we have found over 25 potential novel regulators of this pathway, summarized in Chapters 2 and 4. The top hit is the splicing factor and transcriptional co-activator SNW1. We have further validated it as a regulator of the NF-κB pathway in response to multiple stimuli and in five different cell lines (THP-1, U87, 293T, A549, and U2-OS). SNW1 does not seem to affect general constitutive transcription in THP-1 cells but does seem to repress some transcription programs e.g. CREB and NFE2. SNW1 does not regulate the cytoplasmic part of the NF-κB pathway but does complex with the NF-κB hetero dimer in the nucleus on pathway activation. We have shown that it binds to NF-κB’s transcriptional elongation partner p-TEFb and helps recruit it to the NF-κB nuclear complex that contains RNA Polymerase II. We have also shown that SNW1 loses binding from its splicing complex (SNRNP200, SNRNP220) on NF-κB activation. SNW1 is a unique protein shown to be involved in both splicing and transcription and in the case of NF-κB, its role seems to involve recruitment of p-TEFb for effective transcriptional elongation of NF-κB target genes

Modeling the bending stiffness of point bonded non-woven fabrics

M.S.Prashant Desa