Molecular identification of Saraca asoca from its substituents and adulterants.

ABSTRACT: Saraca asoca (Roxb.) De Wilde is an important medicinal plant from the Western Ghats of India, traditionally used in treatment of various gynecological disorders. Increasing commercial demand and decreasing numbers has resulted in this plant becoming endangered with crude drug materials being extensively substituted/adulterated with other plant species. The present study was undertaken with the objective of development and evaluation of multivariate cluster analysis of ISSR fingerprints against rbcL-based DNA barcodes as tool to understand the relationships and to differentiate common adulterants and substituents from S. asoca. ISSR-based Hierarchical Cluster Analysis was carried out on 41 samples of S. asoca and 5 each of the 5 common substituent/adulterant plants and the clustering patterns were evaluated against DNA-sequence-based barcoding of rbcL region of their plastids. Factorial analysis and Principal Coordinate Analysis revealed distinct groups of genetic pools of respective taxa thereby confirming the utility of ISSR fingerprinting as a useful tool for differentiation between the genuine and the adulterants/substituents. NCBI-BLAST search on DNA barcode rbcL region confirmed the results of ISSR assays. Therefore, our study demonstrated the utility of simple, cost-effective method of ISSR fingerprinting coupled with rbcL barcoding in differentiating this important medicinal plant from its common adulterants/substituents.

Inhibitory effect of polyunsaturated fatty acids on apoptosis induced by Streptococcus pneumoniae in alveolar macrophages.

BACKGROUND & OBJECTIVES: Apoptosis is considered as a major defense mechanism of the body. Multiple pathogens induce macrophage apoptosis as a mode of immune evasion. In earlier studies, n-3 polyunsaturated fatty acids (PUFA) have been reported to be protective against neuronal apoptosis and neuronal degeneration, seen after spinal cord injury. In this study, we tried to evaluate the role of n-3 polyunsaturated fatty acids on the process of macrophage phagocytic activity and apoptosis in mice. METHODS: Mice were divided into three groups (n=60); Group I was fed on sea cod oil; Group II on flaxseed oil supplementation for 9 wk along with standard laboratory chow diet. Group III was fed on standard diet and served as control. After supplementation, phagocytic and apoptotic (morphological staining: acridine orange plus ethidium bromide; H-33342 plus propidium iodide staining and DNA ladder formation) activities of mouse alveolar macrophages were assessed. RESULTS: Alveolar macrophages (obtained from sea cod oil and flaxseed oil fed group mice) showed significant increase in bacterial uptake as well as intracellular killing (P 0.05) of Streptococcus pneumoniae. Significant decrease (P<0.05) in apoptotic cells was observed among alveolar macrophages from sea cod and flaxseed oil fed mice whereas maximum apoptosis was observed in control alveolar macrophages on interaction with bacteria in vitro which was confirmed by DNA laddering. INTERPRETATION & CONCLUSIONS: These findings suggest that dietary supplementation with n-3 polyunsaturated fatty acids to mice led to enhanced phagocytic capability of their alveolar macrophages as well as provided protection against apoptosis upon challenge with S. pneumoniae.

Long-term flaxseed oil supplementation diet protects BALB/c mice against Streptococcus pneumoniae infection.

Intense host immune response to infection contributes significantly to the pathology of pneumococcal pneumonia. Therefore, the regulation of host immune response is critical for the successful outcome of pneumonia in such patients. The aim of the present study was to investigate the effect of n-3 PUFA, i.e. flaxseed oil supplementation for short (4 weeks) as well as long (9 weeks) term, on the course of S. pneumoniae D39 serotype 2 infection in mice. The efficacy of flaxseed oil supplementation was investigated in terms of survival of animals and production of various inflammatory molecules (malondialdehyde, myeloperoxidase, nitric oxide) in the lung homogenate of animals. This was correlated with bacteriological and histopathological parameters. The immunomodulation was studied in terms of cytokines in the lungs following infection with Streptococcus pneumoniae. Results suggest that long-term flaxseed supplementation protected the mice against bacterial colonization of lungs with Streptococcus pneumoniae with reduced histopathological involvement of lung tissue. Moderate pneumonia was observed in supplemented, infected mice compared to severe pneumonia seen in control mice. This was accompanied by decreased inflammatory markers (malondialdehyde, myeloperoxidase, nitric oxide) as the disease progressed. In addition, difference in the levels of pro-inflammatory (TNF-alpha and IL-1beta) and anti-inflammatory (IL-10) cytokines was observed in the flaxseed fed animals. On the contrary, short-term supplementation did not show such an effect on lung colonization.

Induction of resistance to respiratory tract infection with Klebsiella pneumoniae in mice fed on a diet supplemented with tulsi (Ocimum sanctum) and clove (Syzgium aromaticum) oils.

BACKGROUND AND PURPOSE: The impact of diet and specific food groups on respiratory tract infections has been widely recognized in recent years. This study was conducted to study the effect of tulsi (Ocimum sanctum) oil and clove (Syzgium aromaticum) oil on the susceptibility of experimental mice to respiratory tract infection. METHODS: The effect of 2 different regimens of short-term (15 days) and long-term (30 days) feeding with tulsi oil and clove oil on the course of Klebsiella pneumoniae American Type Culture Collection 43816 infection in the lungs of mice was analyzed. The operative mechanisms of lipid peroxidation/nitrite production were studied by estimating their levels in bronchoalveolar lavage fluid (BALF). Bacterial colonization, malondialdehyde (MDA) and nitrite production in BALF, and tumor necrosis factor-alpha level in serum were assessed. RESULTS: The results showed that there was a significant decrease in bacterial colonization after short-term feeding with clove oil compared with the controls (p < 0.05). For tulsi oil-fed mice, the decrease in bacterial load was significant with long-term feeding (p < 0.01). The maximum decrease in MDA levels and increase in nitrite levels were noted with long-term feeding. CONCLUSION: Dietary supplementation with tulsi and clove oils protects against bacterial colonization of the lungs.