In vitro plant regeneration and metabolite profiling of an aromatic medicinal plant Ruta graveolens L. by using GC-MS

Ruta graveolens L. is an endemic plant of the Mediterranean region. It has been used for centuries as a medical preparation and has a variety of roles because of its varied chemical composition. In vitro culture is a useful tool for both multiplication and study of important secondary metabolites. The present study was aimed to develop an effective and reproducible protocol for callus induction and indirect plant regeneration of Ruta graveolens (L.) by using leaf explants and to analyze chemical components present in different extracts of Ruta graveolens. The leaf explants were cultured on MS medium augmented with different combinations and concentrations of auxins and cytokinins for callus induction, shoot multiplication and rooting. The optimum plant growth regulator concentration for callus induction, shoot multiplication and root formation was recorded in MSM+2,4-D(1.5mg/L)+NAA(1.5 mg/L), MSM+ BAP (1.5mg/L) +IBA (1.0 mg/L) and half strength MSM+IBA(0.50 mg/L) respectively. The rooted Plantlets were successfully acclimatized and established in earthen pots. The leaves, stem, roots and callus of Ruta graveolens were extracted by using Acetone and Ethyl acetate solvents followed by volatile compound analysis using GC-MS. The phytochemical assay showed that extracts of Ruta graveolens contain various phytoconstituents having potential bioactivity. The major compounds found were 1, 3-Dioxolane-4-propanol, 2, 2-Dimetheyl-, kokusaginine, Bergaptene, 2 Undecanone, 3-Hexene-2-one, Alpha, - 1- Arabinopyranose, 1, 2:3,4-bis-o, and 1- (1,3-Benzodioxol-5-ylmethyl)-3-Nitro-1 which might be primarily contributing in the biological activity of the plant. The results of this study will make a way for the production of herbal medicines for various ailments by using callus cultures of Ruta graveolens L

Use of Moringa oleifera Flower Pod Extract as Natural Preservative and Development of SCAR Marker for Its DNA Based Identification

The use of Moringa oleifera as natural food preservative has been evaluated in the present study. In addition, for quality assurance, the study has also been focused on the shelf life of product to authenticate the identification of plant by development of DNA based marker. Among the different extracts prepared from flower pods of Moringa oleifera, methanol and aqueous extract exhibited high antibacterial and antioxidant activity, respectively. The high phenolic contents (53.5±0.169 mg GAE/g) and flavonoid contents (10.9±0.094 mg QE/g) were also recorded in methanol and aqueous extract, respectively. Due to instability of bioactive compounds in aqueous extract, methanol extract is considered as potent natural preservative. The shelf life of methanol extract was observed for two months at 4°C under dark conditions. The developed SCAR primers (MOF217/317/MOR317) specifically amplified a fragment of 317 bp from DNA of Moringa oleifera samples collected from different regions of Punjab province of Pakistan. The methanol extract of Moringa oleifera flower pods has great potential to be used as natural preservative and nutraceutical in food industry

Molecular typing of HLA-class II alleles reveals an association with autoantibodies and disease subsets of systemic sclerosis in a North Indian (Kashmir) population

Aim of the work: To identify specific human leukocyte antigen (HLA)-Class II (DRB/DQB1/DPB1) alleles associated with systemic sclerosis (SSc) and to explore their relation with SSc autoantibodies, clinical manifestations, and disease subsets. Patients and methods: HLA-class II alleles (DRB1/DRB3/DRB4/DRB5/DQB1) were determined by DNA typing in 80 SSc cases and 60 matched controls and HLA-DPB1 in 40 SSc patients and 30 controls by allele-specific-polymerase chain reaction with sequence-specific primers (PCR-SSP). Results: The mean age of SSc patients was 36.9 ± 9.4 years; 76 females and 4 males (F:M 19:1) and a disease duration of 5.3 ± 3.3 years, they were 43(53.7%) limited and 37(46.2%) diffuse subtypes. SSc was significantly associated with DRB1*11, DRB1*01, DQB1*04, and DQB1*03*03 in a >4-fold manner, whereas DPB1*04 had a >7-fold increased risk compared to controls. There was a strong association between DRB1*11 (p = 0.04), DQB1*03*03 (p = 0.005), and DPB1*13 (p = 0.009) with anti-topoisomerase I (anti-topoI) whereas the frequency of DRB1*01 (p < 0.0001) was increased in patients with anti-centromere (ACA) positive SSc compared those negative (56% vs 25%; p < 0.0001). DRB1*03, DRB1*15, and DQB1*03*01 were SSc protective alleles in patients with positive ACA. Anti-topo I was associated with interstitial lung disease (ILD) (p < 0.01), whereas ACA with pulmonary arterial hypertension (PAH) (p = 0.01) and protection against ILD (p < 0.001). In addition, HLA-DRB1*03, DQB1*03*01and DPB1*03 were more frequent in patients with ILD than in patients without. Conclusion: Associations between specific HLA-class II alleles with certain SSc-specific autoantibodies (anti-topo I and ACA) were identified. Specific HLA associations with clinical and serological subtypes could serve as biomarkers of disease severity and progression in SSc