'In vitro' antiviral activity of neem (Azadirachta indica. A. Juss) leaf extract against group B coxsackieviruses

The antiviral and virucidal effect of methanolic extract fraction of leaves of neem (Azadirachta indica A. Juss) (NCL-11) was studied regarding its activity and possible mechanism of action against Coxsackie B group of viruses. NCL-11 inhibited plaque formation in 6 antigenic types of Coxsackie virus B at a concentration of 1000 micrograms/ml at 96 hrs. 'in vitro'. Additionally virus inactivation, yield reduction and effect of time of addition assays suggested that NCL-11 was most effective against coxsackie virus B-4 as a virucidal agent besides interfering at an early event of its replicative cycle. The evidence suggested that presence of a battery of compounds besides flavonoids, triterpenoids and their glycosides in NCL-11 have antiviral action for coxsackie B group of viruses 'in vitro.' The minimal inhibitory concentrations were not toxic to Vero (African green monkey kidney), cells; subtoxic concentration was 8,000 micrograms/ml and cytotoxic concentration 10,000 micrograms/ml, which was confirmed by trypan blue dye exclusion test.     

Meta-analysis on pharmacological therapies in the management of xerostomia in patients with Sjogren’s syndrome

Introduction: Sjogren’s syndrome is an immunologic disorder, characterized by symptoms of dry mouth and dry eyes. Management of xerostomia is more difficult and challenging, various pharmacologic agents have been tried and evaluated in the management of xerostomia in these patients, but the results were inconsistent and variable. Hence, the present study is aimed at evaluation and comparison of different pharmacological agents in the management of xerostomia in patients with Sjogren’s syndrome.

Materials and methods: A meta-analysis of case–control studies was conducted on pharmacological management of xerostomia in patients with Sjogren’s Syndrome and the collected data are subjected to exclusion and inclusion criteria and standard mean difference (SMD), ODD’s ratio and confidence intervals (95% CI) were calculated by Review Manager software using fixed and random effects model from the data of five studies.

Results: Both objective response and subjective response evaluation favored experimental group suggesting an increase in unstimulated salivary flow rate using pharmacological agents. Interferon alpha 150?IU three times daily had a good effect in increasing the unstimulated whole saliva flow rate with SMD 2.72 at 95% CI [2.43, 3.00] p?<?.00001. Cevimeline vs placebo showed good response with ODDS ratio 2.74 at 95% CI [1.58, 4.76] p?=?.0003.

Conclusion: Interferon – α 150?IU thrice daily was proven to be effective in increasing salivary flow rate and also has an advantage of disease modification in SS patients attributing to its immunomodulatory action. Cevimeline 30?mg thrice daily also had a considerable therapeutic effect in SS patients compared to Pilocarpine.     

In vitro antiviral activity of bael (Aegle marmelos Corr) upon human coxsackieviruses B1-B6

The in-vitro antiviral activity of a series of compounds in samples extracted from various parts of the Indian holy tree, Bael (Aegle marmelos corr.) were evaluated for their efficacy against human coxsackieviruses B1-B6. The inhibitory concentrations (IC50) for leaves (L1 and L2) stem and stem bark (S1, S2, S3 and S4) fruit (F1 and F2micro) root and root bark (R1 and R2) and pure compound, the marmelide were 1000 microg/ml (for L1 and L2), 1000 microg/ml (for S1, S2, S3 and S4), 1000 microg/ml (for F1) and 500 microg/ml (for F2) 250 microg/ml (for R1) and 500 microg/ml (for R2) and 62.5 microg/ml for marmelide respectively by plaque inhibition assay at 96 hrs. On the other hand, the corresponding value for Ribavirin, a standard antiviral drug, was 2000 microg/ml for the same viruses at the same time period. These concentrations did not exhibit any toxicity to Vero cells, the host subtoxic concentrations were 5000 microg/ml for leaf and stem fractions 2000 microg/ml for fruit fractions 500 and 1000 microg/ml for root fractions 250 microg/ml for marmelide and 2000 microg/ml for Ribavirin. The cytotoxic concentrations were 8000 microg/ml for leaf and stem compounds 4000 mg/ml for fruit; 1000 microg/ml and 2000 microg/ml for root 500 microg/ml for marmelide and 4000 microg/ml for ribavirin at 96 hrs. These were also confirmed by trypan blue dye exclusion test and further passaging of cells. Additionally pretreatment of host cells, virus inactivation, yield reduction and effect of time of addition assays against coxsackievirus B3 suggested that marmelide was most effective as a virucidal agent besides interfering at early events of its replicative cycle like adsorption, penetration, at various steps in single cycle growth curve and effect of time of addition.     

Homez,a homeobox leucine zipper gene specific to the vertebrate lineage

This work describes a vertebrate homeobox gene, designated Homez (homeodomain leucine zipper-encoding gene), that encodes a protein with an unusual structural organization. There are several regions within Homez, including three atypical homeodomains, two leucine zipper-like motifs, and an acidic domain. The gene is ubiquitously expressed in human and murine tissues, although the expression pattern is more restricted during mouse development. Genomic analysis revealed that human and mouse genes are located at 14q11.2 and 14C, respectively, and are composed of two exons. The zebrafish and pufferfish homologs share high similarity to mammalian sequences, particularly within the homeodomain sequences. Based on homology of homeodomains and on the similarity in overall protein structure, we delineate Homez and members of ZHX family of zinc finger homeodomain factors as a subset within the superfamily of homeobox-containing proteins. The type and composition of homeodomains in the Homez subfamily are vertebrate-specific. Phylogenetic analysis indicates that Homez lineage was separated from related genes >400 million years ago before separation of ray- and lobe-finned fishes. We apply a duplication-degeneration-complementation model to explain how this family of genes has evolved.     

Detection of Coxsackievirus B 4 antibodies in cases of suspected encephalopathy by microneutralization test

A retrospective study of serum and cerebrospinal fluid (CSF) samples collected from suspected viral encephalitis and encephalopathy cases was carried-out and it included 100 CSF and 89 serum samples from Goa, collected during 1990-1994. These samples which were negative for antibodies to Japanese encephalitis (JE), West Nile (WN), Dengue-2 (DN-2) and herpes viruses, were tested for Coxsackievirus B 4 specific antibodies by 'in vitro' microneutralization technique along with 80 negative control serum samples. Out of 189 specimens (100 CSF and 89 serum), 23 CSF and 41 serum samples were positive for Coxsackievirus B 4 neutralizing antibodies. Antibody profile seemed to be IgG as revealed by mercaptoethanol treatment. The presence of neutralizing antibodies to coxsackievirus B 4 with titres as high as 1:512 in 8 CSF and 19 serum samples seemed to be suggestive of viral meningitis due to Cox B-4 viruses.     

GTF2IRD2 is located in the Williams-Beuren syndrome critical region 7q11.23 and encodes a protein with two TFII-I-like helix-loop-helix repeats

Williams-Beuren syndrome (also known as Williams syndrome) is caused by a deletion of a 1.55- to 1.84-megabase region from chromosome band 7q11.23. GTF2IRD1 and GTF2I, located within this critical region, encode proteins of the TFII-I family with multiple helix-loop-helix domains known as I repeats. In the present work, we characterize a third member, GTF2IRD2, which has sequence and structural similarity to the GTF2I and GTF2IRD1 paralogs. The ORF encodes a protein with several features characteristic of regulatory factors, including two I repeats, two leucine zippers, and a single Cys-2/His-2 zinc finger. The genomic organization of human, baboon, rat, and mouse genes is well conserved. Our exon-by-exon comparison has revealed that GTF2IRD2 is more closely related to GTF2I than to GTF2IRD1 and apparently is derived from the GTF2I sequence. The comparison of GTF2I and GTF2IRD2 genes revealed two distinct regions of homology, indicating that the helix-loop-helix domain structure of the GTF2IRD2 gene has been generated by two independent genomic duplications. We speculate that GTF2I is derived from GTF2IRD1 as a result of local duplication and the further evolution of its structure was associated with its functional specialization. Comparison of genomic sequences surrounding GTF2IRD2 genes in mice and humans allows refinement of the centromeric breakpoint position of the primate-specific inversion within the Williams-Beuren syndrome critical region.