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To design novel antimicrobial peptides by utilizing the sequence of the human host defense protein, chemerin, a seven-residue amphipathic stretch located in the amino acid region, 109-115, was identified, which possesses the highest density of hydrophobic and positively charged residues. Although this 7-mer peptide was inactive toward microorganisms, its 14-mer tandem repeat (Chem-KVL) was highly active against different bacteria including methicillin-resistant Staphylococcus aureus, a multidrug-resistant Staphylococcus aureus strain, and slow- and fast-growing mycobacterial species. The selective enantiomeric substitutions of its two l-lysine residues were attempted to confer cell selectivity and proteolytic stability to Chem-KVL. Chem-8dK with a d-lysine replacement in its middle (eighth position) showed the lowest hemolytic activity against human red blood cells among Chem-KVL analogues and maintained high antimicrobial properties. Chem-8dK showed in vivo efficacy against Pseudomonas aeruginosa infection in BALB/c mice and inhibited the development of resistance in this microorganism up to 30 serial passages and growth of intracellular mycobacteria in THP-1 cells.

Aim: This study is aimed to compare and evaluate the changes in the microflora in immediate and delayed placed implants. Materials and Methods: In this study, the implant site sample was taken and assessed during different phases of treatment for delayed and immediate implants. They were looked for Streptococcus, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, and Fusobacterium nucleatum. Results: The results showed that Streptococci were found in a higher number in all the phases of the treatment. The presence of pathogenic organisms such as P. gingivalis and Fusobacterium, in considerable amounts, was seen in both the groups. Conclusion: Thus, we conclude that implant mode of placement, delayed or immediate placement does not alter the flora of the oral cavity. Organisms present remains the same in all the phase of the treatment. To prevent the disease, one must continuously monitor the implant, with the increasing age changes, the microflora is continually changing in the oral cavity. The periodontal health should be assessed before the placement of the implant, followed by follow-ups after a set period for a better prognosis.

A technology for systemic and repeated administration of osteogenic factors for orthopedic use is an unmet medical need. Lactoferrin (∼80 kDa), present in milk, is known to support bone growth. We discovered a lactoferrin-mimetic peptide, LP2 (an 18-residue fragment from the N-terminus of the N-lobe of human lactoferrin), which self-assembles into a nano-globular assembly with a β-sheet structure in an aqueous environment. LP2 is non-hemolytic and non-cytotoxic against human red blood cells and 3T3 fibroblasts, respectively, and appreciably stable in the human serum. LP2 through the bone morphogenetic protein-dependent mechanism stimulates osteoblast differentiation more potently than the full-length protein as well as the osteoblastic production of osteoprotegerin (an anti-osteoclastogenic factor). Consequently, daily subcutaneous administration of LP2 to rats and rabbits with osteotomy resulted in faster bone healing and stimulated bone formation in rats with a low bone mass more potently than that with teriparatide, the standard-of-care osteogenic peptide for osteoporosis. LP2 has skeletal bioavailability and is safe at the 15× osteogenic dose. Thus, LP2 is a novel peptide that can be administered systemically for the medical management of hard-to-heal fractures.

Mitogen-Activated Protein Kinase (MAPK)-Activated Protein Kinase 2 (MK2), downstream to p38MAPK (p38mitogen-activated protein kinase), regulates cellular inflammation and proliferation. So far, the role of MK2 has been studied in many cardiovascular diseases, but it remains unexplored in pulmonary hypertension (PH). Therefore, to investigate the role of MK2 in the PH pathogenesis, human pulmonary artery smooth muscle cells were exposed to hypoxia (1% O 2 ) for 72 hours, and MK2 was inhibited by siRNA. We observed significantly increased MK2 expression, inflammatory cytokines, proliferation, mitochondrial dysfunction, and apoptosis resistance in hypoxic human pulmonary artery smooth muscle cells, which were reversed by treatment with MK2 siRNA. For in vivo studies, male Sprague Dawley rats were treated with monocrotaline (60 mg/kg, SC, once) to induce PH. To inhibit MK2, a peptide MMI-0100 (40 μg/kg, IP daily, 5 weeks for preventive and 3 weeks for curative study) was administered. MMI-0100 treatment decreased right ventricle pressure and hypertrophy, hallmarks of PH, in both preventive and curative study. MMI-0100-treated rats showed better cardiac functions as revealed by 2-dimensional echocardiography study. Furthermore, MMI-0100 reversed pulmonary vascular remodeling and improved pulmonary vascular relaxation in monocrotaline-treated rats. Finally, the above results were confirmed in MK2 knockout mice. MK2 knockout mice, received 600 mg/kg monocrotaline, subcutaneous weekly for 5 weeks, failed to develop PH and showed no increase in right ventricle pressure and hypertrophy. This study, therefore, proved that MK2 is involved in PH, and its inhibition may be a novel target for PH treatment.

Chrysanthemum is the next only to rose in importance among the flower crops in the world. In India it has been recognised as fifth commercially important potent flower crop. It is versatile, it can be planted in pots, used for garland making and also as cut flower for flower arrangement. In India, large flowered varieties are grown for exhibition purpose while small flowered varieties are grown for cut flower, making garland, wreaths, veni and religious offerings. Apart from their ornamental flower, Chrysanthemum is a good source of nutrients, vitamins and minerals. The most important chemical extracts of Chrysanthemum include flavonoids, betaine, choline and vitamin B1. Because elite cultivars are typically propagated vegetatively, the risk of disseminating fungal, bacterial, phytoplasmal and viral pathogens is high. A number of viruses are known to infect chrysanthemum that cause benign, stunt, chlorosis, mosaic symptoms, and mottle. Consequently, this has led to qualitative and quantitative losses, resulting in serious problems in chrysanthemum production worldwide. Some viruses which affect the production of chrysanthemum are: - Chrysanthemum virus B (CVB), Chrysanthemum stem necrosis virus (CSNV), Chrysanthemum stunt viroid (CSVd), Cucumber mosaic virus (CMV), Tomato aspermy virus (TAV), Tomato spotted wilt virus (TSWV), Potato virus X and Begomovirus. Management of viral diseases is generally similar to those used for other pathogens, except that as yet chemicals find little application in viral diseases, although they may be used for vectors. Moreover, vectors involved in viral diseases have also complicated the problem in their management. The production of flowers and ornamental plants has been continuously growing due to improvement in market structure, growth in population’s and producers’ purchasing power, diversification of species, diffusion of new production technologies and professionalization of the members of the Brazilian production chain. Factors, such as, number of plants, genetics and environmental, are determinant for the production of floral stems with quality. This quality can be evaluated using the length, stiffness and healthy floral stem, diameter and degree of inflorescence opening.

From the functional standpoint, broadly, three kinds of functional DNA sequences exist: one that encodes proteins, second that encodes only RNA (noncoding DNA), and third that does not transcribe at all (Noncoding DNA) historically people have paid attention to the protein-coding genes. For the last two-and-a-half decades, the noncoding RNA has taken center stage. However, the role of noncoding DNA (the dark matter of genome) is still largely unknown. We asked a simple question: How did nature decide to allocate protein-coding and RNA-coding jobs to a specific set of sequences? Did she sample all possibilities in the neighborhood of protein-coding genes, retaining good results (protein- and RNA-encoding genes), retiring not-so-relevant results (pseudogenes), and leaving some genome sequences untouched (nonexpressive genome) To answer these, we expressed some noncoding intergenic sequences from Escherichia coli into functional proteins. The success of this pilot experiment led us toward building a knowledge base that predicts the outcome of making user-defined genes from nonexpressive DNA sequences. In this chapter, the idea of lab-made genes shall be presented from the original concept to its present state where novel therapeutic molecules against cancer, infectious diseases, and neurodegenerative diseases have been found. We call this new platform “FLAGSHIP”—a novel drug discovery platform originating from the dark matter of the genome.

The effect of simulated physiological oocyte maturation on the developmental competence, reactive oxygen species production and apoptosis rate of in vitro produced goat embryos were studied in the present experiment. Oocytes and spermatozoa were recovered from ovaries and epididymis, respectively, procured from a local small animal abattoir. The oocytes aspirated from the ovaries were allocated into two groups, control (subjected to routine in vitro maturation, fertilization and culture) and simulated physiological oocyte maturation (SPOM) group (subjected to prematuration, followed by routine in vitro maturation, fertilization and culture). The SPOM group showed a significantly (p 0.05) higher maturation and blastocyst rates (90.60 ± 0.46% and 29.09 ± 2.59%, respectively) as compared to the control group (85.29 ± 0.98% and 24.09 ± 1.08%). The intensity of reactive oxygen species of the embryos in the control group (14.98 ± 0.83 pixels/embryo) was significantly (p 0.05) higher than the SPOM group (9.60 ± 0.76 pixels/embryo). The apoptosis rate was also significantly (p 0.05) higher in the embryos of the control group (9.18 ± 1.07%) as compared to the SPOM group (5.71 ± 0.90%). In conclusion, the simulated physiological oocyte maturation system significantly increases the developmental competence of the oocytes and decreases the intensity of reactive oxygen species and embryonic apoptosis in abattoir derived goat embryos.

In the present study effect of asiaticoside, on healing of skin wounds in Cirrhinus mrigala is reported. Skin wound, approx. 2mm in diameter was excised using sterile disposable biopsy punch. Immediately after infliction of the wound, epidermis from wound edge starts migrating as thin sheet toward wound gap. Fronts of migrating epidermis gradually advance, and results in complete epithelialization of wound. Experiments were conducted for 30days and fishes were divided into control, sham, vehicle control and asiaticoside treated groups. Immunohistochemical localization of proliferating cell nuclear antigen positive cells indicating cellular proliferation and caspase 3 positive cells reflecting apoptosis was carried out and their density at different post wound intervals in each fish group was analyzed. Significant increase in cellular proliferation as well as decrease in apoptosis in both epidermis and dermis in fish treated with asiaticoside compared to sham and vehicle control fish is observed at different intervals of wound repair. This suggests that in treated group healing of skin wounds in fish is enhanced than in sham and vehicle control groups. Asiaticoside treatment in healing of skin wounds would greatly be beneficial to fish farmers as it could protect fish from invasion of pathogens and check fish mortality.

Present scanning electron microscope study, reports healing of excised skin wounds in Cirrhinus mrigala. Healing process of wounds, inflicted on head skin, using biopsy punch was observed at intervals-0 hour (h), 1, 2, 4, 6, 12 h, 1 day (d) 2 and 4 d. Accumulation of mucus in wound region within 1h after infliction of wound has been considered an immediate measure to provide protection to injured skin from microbial invasion and other external environmental hazards. On infliction of wound, mobilization of epithelial cells at wound edge is associated with disturbance of coaptive relationship of epithelial cells till original coaptive stability is reached. At 6-12 h appearance of epidermal ridge in region of contact of migrating fronts is due to piling up of epithelial cells. This is associated with cessation of migration of epithelial cells and their simultaneous continual arrival in the region. Speedy epithelialization of skin wounds in C. mrigala like in other fishes, compared to that of mammals and other higher vertebrates, is possibly facilitated owing to surrounding wet external environment. Microridges in initial stages of wound healing appear fragmented without particular orientation. Further, epithelial cells in epithelium in wound region and in region adjacent to wound elongate. These changes are associated with the stretching of epithelial cells indicating their streaming and migration, toward wound. Presence of superficial neuromasts, smallest functional units of lateral line system, a hydrodynamic sensory system, has been associated with important functional significance in fish.

The antioxidative, cardioprotective, and anticancer potential of extracts of the edible lichens Everniastrum cirrhatum and Parmotrema reticulatum were evaluated. Acetone extracts of P. reticulatum exhibited nitric oxide scavenging and anti-lipid peroxidation in the range of 35.0–97.0%. IC50 values of extracts for angiotensin-converting enzyme and HMG-CoA reductase inhibition were observed in the range of 97–375 µg.mL–1 and 89–118 µg.mL–1, respectively, against standard inhibitors captopril (32 µg.mL–1) and pravastatin (26 µg.mL–1); methanol extract of P. reticulatum displayed the highest cytotoxicity, reducing HCT-116 cell viability to 40%, suggesting potential use of these lichens as nutraceuticals.

Surface ultrastructure of the gills of the angler catfish Chaca chaca was investigated to unravel the adaptive modifications associated with the feeding ecology of the fish. The fish is often found in mud or in soft substrates where they remain buried both for protection and to feed. Gill rakers present on the gill arch in most fish species are absent in this fish. The absence of gill rakers are associated with the feeding habit of the fish and is considered to facilitate the swallowing of captured prey smoothly without any hindrance. Highly corrugated surface of the gill arch and gill filaments could be associated to retain water/mucus to prevent dessicassion of the fish. Papillae like epithelial protuberances each bearing a taste bud at its summit toward the pharyngeal side of the gill arch is associated with the sorting of the food. Large number of mucous goblet cells on the gill arch epithelium are considered to secret copious mucus to lubricate the prey for easy swallowing. In C. chaca the gill septa between gill filaments are reduced. This could enhance the flexibility and permit the free movement of the gill filaments. Extensive secondary lamellae and infrequent mucous goblet cells on secondary lamellae are associated to increase the surface area to enhance efficiency of gaseous exchange.

In this investigation, the effect of dietary administration of curcumin on the healing of skin wound in fish, Labeo rohita, has been reported. Fish were divided into three groups: control group (fish without skin wound), sham group (fish with skin wound without curcumin treatment) and curcumin-treated group (fish with skin wound and subjected to dietary administration of 1% curcumin). Experiments were conducted for 30 days to assess the healing of skin wounds at different time intervals using scanning electron microscopy, histology, and mucopolysaccharide and enzyme histochemistry. In the curcumin-treated group, healing of skin wounds was found to be enhanced than in the sham group as indicated by early restoration of morphology of the surface layer of epithelial cells; the density of the mucous goblet cells; the density of club cells in epidermal layer; and early granular tissue formation, collagen deposition and tissue remodelling in dermal layer. Furthermore, peroxidase and catalase enzyme activity showed increased endogenous defence system in the curcumin-treated group compared with the sham group. It could be concluded that dietary administration of curcumin is beneficial in rapid healing of skin wounds in fish. Early healing of wounds could be considered to prevent the invasion of pathogens and to maintain the integrity of the surrounding tissue.

MicroRNAs (miRNAs) are small non-coding RNA molecules of around 22 nucleotides in length. They are crucially involved in the post transcriptional regulation and thus play a significant role in the modulation of different diseases. Several studies have suggested that miRNA expression is dysregulated in various cancers through different mechanisms and the dysregulated miRNA in return affects different cancer hallmarks including cell proliferation, cell death suppression, metastasis and angiogenesis. Compilation of the available miRNA data can be a stimulator for proper understanding of the correlation between the miRNA expression and cancer progression. In this review, we have focussed on the role of miR-760 in the progression of different cancer. MicroRNA-760 (miR-760) has been found to be down regulated in various cancers, thus it can be utilized as a possible prognostic marker for cancer detection. Here, we have tried to fill a gap regarding the role of miR-760 in relation to cervical cancer also. Moreover, unravelling the role of miR-760 in different cancers will enlighten the researchers with proper understanding of biology of miR-760 in regulation of different cancers.

Anthraquinones (AQs) of the Cassia occidentalis (CO) seeds like Rhein, Emodin, Aloe-emodin, Chrysophanol, Physcion are known to contribute to the etiology of hepatomyoencephalopathy in children. The present study was carried out to investigate the binding affinity of these AQs with serum albumin, as this protein is mainly involved in the bio-distribution of xenobiotics. Initially, molecular docking was carried for the five AQ ligands with bovine serum albumin (BSA) both by AUTODOCK and CDOCKER docking tools. Subsequently, the binding affinity and mechanism of interaction of these AQs with BSA was studied by fluorescence and UV–visible spectroscopy. Additionally, in vitro cytotoxicity studies of these AQs were carried out in rat primary hepatocytes and HepG2 cells. Results both from molecular docking and fluorescence spectroscopic studies suggest that the interaction energy and binding affinity of above AQs with BSA is in the following order, i.e. Rhein > Emodin > Aloe-emodin > Chrysophanol > Physcion. Interestingly, the observed cytotoxicity of the five AQs in rat primary hepatocyte and HepG2 cells were found to fall in line with the above order. Rhein showing highest toxicity among the AQs showed highest binding affinity to BSA, whereas chrysophanol & physcion, being less toxic exhibited minimal affinity to BSA. Hence, these studies indicate that toxicity of AQs of CO seeds are directly proportional to the protein binding affinity, which may be responsible for the death of children and cattle fed of CO seeds.

Histopathological changes and alterations in the activity of certain metabolic and antioxidant enzymes were analyzed in the head skin of Labeo rohita, exposed to sublethal test concentrations of the azo dye, Eriochrome black T for 4 days, using 24 h renewal bioassay method. Hypertrophied epithelial cells, increased density of mucous goblet cells, and profuse mucous secretion at the surface were considered to protect the skin from toxic impact of the azo dye. Degenerative changes including vacuolization, shrinkage, decrease in dimension, and density of club cells with simultaneous release of their contents in the intercellular spaces were associated to plug them, preventing indiscriminate entry of foreign matter. On exposure of fish to the dye, significant decline in the activity of enzymes—alkaline phosphatase, acid phosphatase, carboxylesterase, succinate dehydrogenase, catalase, and peroxidase—was associated with the binding of dye to the enzymes. Gradual increase in the activity of lactate dehydrogenase was considered to reflect a shift from aerobic to anaerobic metabolism. On transfer of azo dye exposed fish to freshwater, skin gradually recovers and, by 8 days, density and area of mucous goblet cells, club cells, and activity of the enzymes appear similar to that of controls. Alteration in histopathology and enzyme activity could be considered beneficial tool in monitoring environmental toxicity, valuable in the sustenance of fish populations.

RuBisCo activase (Rca) is a catalytic chaperone involved in modulating the activity of RuBisCo (key enzyme of photosynthetic pathway). Here, we identified eight novel transcripts from wheat through data mining predicted to be Rca and cloned a transcript of 1.4 kb from cv. HD2985, named as TaRca1 (GenBank acc. no. KC776912). Single copy number of TaRca1 was observed in wheat genome. Expression analysis in diverse wheat genotypes (HD2985, Halna, PBW621 and HD2329) showed very high relative expression of TaRca1 in Halna under control and HS-treated, as compared to other cultivars at different stages of growth. TaRca1 protein was predicted to be chloroplast-localized with numerous potential phosphorylation sites. Nothern blot analysis showed maximum accumulation of TaRca1 transcript in thermotolerant cv. during mealy-ripe stage, as compared to thermosusceptible. Decrease in the photosynthetic parameters was observed in all the cultivars, except PBW621 in response to HS. We observed significant increase in the Rca activity in all the cultivars under HS at different stages of growth. HS causes decrease in the RuBisCo activity; maximum reduction was observed during pollination stage in thermosusceptible cvs. as validated through immunoblotting. We observed uniform carbon distribution in different tissues of thermotolerant cvs., as compared to thermosusceptible. Similarly, tolerance level of leaf was observed maximum in Halna having high Rca activity under HS. A positive correlation was observed between the transcript and activity of TaRca1 in HS-treated Halna. Similarly, TaRca1 enzyme showed positive correlation with the activity of RuBisCo. There is, however, need to manipulate the thermal stability of TaRca1 enzyme through protein engineering for sustaining the photosynthetic rate under HS – a novel approach towards development of ‘climate-smart’ crop.

ObjectiveTo investigate the antihyperglycemic and antihyperlipidemic properties of hydroalcoholic extract of fruits of Sapindus mukorossi Gaerten and its beneficial effect on haematological parameters with histopathological analysis in streptozotocin induced diabetic rats.MethodsSapindus mukorossi fruits extract (250 and 500 mg/kg body weight) and standard drug glybenclamide (0.5 mg/kg body weight) were administered to diabetic rats. Effect of extract on hyperglycemia, hyperlipidemia and hematological parameters was studied in diabetic rats. Histopathological changes in diabetic rat pancreas were also observed after extract and glybenclamide treatment.ResultsDaily oral administration of Sapindus mukorossi fruits extract (250 and 500 mg/kg body weight) and glybenclamide for 20 days showed beneficial effects on blood glucose level (P

ObjectiveTo evaluate the hepatoprotective potential of ethanolic (50%) extract of Ziziphus oenoplia (L.) Mill (Z. oenoplia) root against isoniazid (INH) and rifampicin (RIF) induced liver damage in animal models.MethodsFive groups of six rats each were selected for the study. Ethanolic extract at a dose of 150 and 300 mg/kg as well as silymarin (100 mg/kg) were administered orally once daily for 21 d in INH + RIF treated groups. The serum levels of glutamic oxaloacetic transaminase (SGOT), glutamate pyruvate transaminase (SGPT), alkaline phosphatase (SALP), and bilirubin were estimated along with activities of superoxide dismutase, catalase, glutathione S-transferase, glutathione peroxidase, and hepatic melondialdehyde formation. Histopathological analysis was carried out to assess injury to the liver.ResultsThe considerably elevated serum enzymatic activities of glutamic oxaloacetic transaminase, glutamate pyruvate transaminase, alkaline phosphatase and bilirubin due to INH + RIF treatment were restored towards normal in a dose dependent manner after the treatment with ethanolic extract of Z. oenoplia roots. Meanwhile, the decreased activities of superoxide dismutase, catalase, glutathione S-transferase and glutathione peroxidase were also restored towards normal dose dependently. In addition, ethanolic extract also significantly prevented the elevation of hepatic melondialdehyde formation in the liver of INH + RIF intoxicated rats in a dose dependent manner. The biochemical observations were supplemented with histopathological examination of rat liver sections.ConclusionsThe results of this study strongly indicate that ethanolic extract of Z. oenoplia has a potent hepatoprotective action against INH + RIF induced hepatic damage in rats.

Objective: To evaluate the hepatoprotective potential of ethyl acetate fraction of Rhododendron arboreum (Family: Ericaceae) in Wistar rats against carbon tetrachloride (CCl 4 )-induced liver damage in preventive and curative models. Materials and Methods: Fraction at a dose of 100, 200, and 400 mg/kg was administered orally once daily for 14 days in CCl 4 -treated groups (II, III, IV, V and VI). The serum levels of glutamic oxaloacetic transaminase (SGOT), glutamate pyruvate transaminase (SGPT), alkaline phosphatase (SALP), γ-glutamyltransferase (γ -GT), and bilirubin were estimated along with activities of glutathione S-transferase (GST), glutathione reductase, hepatic malondialdehyde formation, and glutathione content. Result and Discussion: The substantially elevated serum enzymatic activities of SGOT, SGPT, SALP, γ-GT, and bilirubin due to CCl 4 treatment were restored toward normal in a dose-dependent manner. Meanwhile, the decreased activities of GST and glutathione reductase were also restored toward normal. In addition, ethyl acetate fraction also significantly prevented the elevation of hepatic malondialdehyde formation and depletion of reduced glutathione content in the liver of CCl 4 -intoxicated rats in a dose-dependent manner. Silymarin used as standard reference also exhibited significant hepatoprotective activity on post-treatment against CCl 4 -induced hepatotoxicity in rats. The biochemical observations were supplemented with histopathological examination of rat liver sections. The results of this study strongly indicate that ethyl acetate fraction has a potent hepatoprotective action against CCl 4 -induced hepatic damage in rats.

Cytochromes are membrane-bound hemoproteins responsible for the generation of ATP via the electron transport system to fuel the metabolic processes of the organism for their growth. This study reports the properties of cytochromes present in the isolated lichenized fungi of the cultured lichen Usnea ghattensis under optimized conditions. The fungal partner of the cultured lichen Usnea ghattensis contains a, b and c types of cytochromes. The concentrations of a, b and c type cytochromes were found to be significantly high (0.0967, 0.0900, and 0.1030 mM/mg protein, respectively) in the isolated fungal symbiont of cultured lichen grown in malt-yeast extract medium supplemented with 0.01 mol/l sucrose and 0.01 mol/l polyethylglycol. The results suggest that supplementation of additional carbon sources may play a role in optimizing the growth via activating the cytochrome respiratory system in lichenized fungi.

This article has been retracted at the request of the editors and authors. Please see Elsevier Policy on Article Withdrawal ( http://www.elsevier.com/locate/withdrawalpolicy ). Reason: This paper appears to be very similar to another paper by the same authors which was published about the same time: “Tissue Culture of Some Lichens and Screening of Their Antioxidant, Antityrosinase and Antibacterial Properties”, B.C. Behera, Neeraj Verma, Anjali Sonone and Urmila Makhija, Phytother. Res., 21 (2007) 1159–1170. The similarity is too great to justify two primary publications.

Lichen species Usnea ghattensis, Heterodermia podocarpa, Arthothelium awasthii and Parmotrema tinctorum have been cultured in vitro and screened for their antioxidant, antimicrobial and antityrosinase potential using different assay systems. The methanol extract of lichens showed antioxidant, antimicrobial and antityrosinase activities according to the order Usnea ghattensis > Arthothelium awasthii > Heterodermia podocarpa > Parmotrema tinctorum. The IC(50) values for the antioxidant and antityrosinase activities of U. ghattensis and A. awasthii were less than or equivalent to that of standard antioxidants and tyrosinase inhibitors. The methanol extracts of the lichen cultures Usnea ghattensis and Arthothelium awasthii were effective against Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis and Staphylococcus aureus. The minimum inhibitory concentration (MIC) of the extract was found to be between 5 and 10 microg extract/mL. The results suggested that the extract of lichen cultures, U. ghattensis and A. awasthii, could be of use as an easily accessible source of natural antioxidants, tyrosinase inhibitory and antimicrobial properties as possible food supplements or in the pharmaceutical industry.

Lichens are source of natural bioactive compounds which are traditionally used to cure a variety of ailments.The objective of this study is to assess free radical scavenging, prolyl endopeptidase inhibitory (PEPI), and antimicrobial potential of a high altitude lichen species Cetrelia olivetorum (Nyl.) W. L. Culb.C. F. Culb (Parmeliaceae).Lichen C. olivetorum has been cultured in vitro, and optimized culture conditions were implemented in bioreactor to obtain high quantity of biomass for the study of radical scavenging, PEPI, and antimicrobial activities. Radical scavenging activity of methanol extract of Cetrelia olivetorum (MECO) was tested at 100 µg/mL, PEPI activity at 25 and 50 µg/mL, and antimicrobial activity at 5, 25, 50, and 100 µg/mL conc. All the biological activities of natural thallus extract and its derived culture extract were evaluated spectrophotometrically.Murashige and Skoog medium supplemented with 3% glucose and 100 ppb indole-3-butyric acid (IBA) supported biomass growth at flask level and yielded 5.095 g biomass in bioreactor. MECO of both the cultured and the natural lichen exhibited half inhibiting concentration (IC50) for radical scavenging activities in the range of 50-60 µg/mL, whereas the IC50 value of standard antioxidants was found to be in the range of 12-29 µg/mL. The IC50 value of lichen extract for PEPI activity was 144-288 µg/mL, whereas the IC50 value of standard prolyl endopeptidase inhibitor, Z-pro-prolinal, was 57.73 µg/mL. As far as the antimicrobial activity of MECO is concerned, minimum inhibitory concentration (MIC) value of lichen extracts against tested microorganisms was obtained in the range of 50-104 µg/mL and found to be more effective than commercially available standard erythromycin.Murashige and Skoog medium containing IBA was found to be suitable for maximum biomass production of C. olivetorum under bioreactor conditions. The cultured lichen biomass extract also showed antioxidant, PEPI, and antimicrobial potential.The present study indicates therapeutic potential of Himalayan lichen C. olivetorum against neurodegenerative diseases owing to its radical scavenging, PEPI, and antimicrobial activities. Further, the result encourages its commercial exploitation through mass culture for production of its bioactive components and their use in pharmaceutical and nutraceutical industries.

In the world, lichens are slowest growing symbiotic organism associations between fungi and a photosynthetic alga and/or cyanobacteria. Lichens produce a wide array of secondary metabolites which are unique and forms under lichenized conditions. Secondary metabolites of lichens demonstrated significant inhibition of various biological activities at very low concentrations. Although lichens are reservoir for various biologically active compounds, only few lichens and their compounds have been tested for their biological significance and still there are many more yet to be tested. Hence, there is clearly an urgent need for expanding research in this area including in-depth studies of those compounds which have shown promising results. A strong focus is also needed on the most promising lichen-based drug therapies followed by large scale production of the best of those compounds. One of the main issues related to the limited use of lichen compounds in modern medicine is the slow growth rate of lichen thalli which remains as a challenge to enhance their biomass with in vitro culture. Advance research in the field of cultivating lichens and their symbionts will enable the mass production of lichen substances and their pharmaceutical applications. The present review on tissue culture of lichens of various groups, biosynthesis of their unique secondary compounds, physiological conditions required for their synthesis, and biological activities will significantly contribute to the present knowledge in the field of experimental lichenology and will also attract the attention of industry/society/nation as a whole.

A viral disease was identified on geraniums (Pelargonium spp.) grown in a greenhouse at the Institute of Himalayan Bioresource Technology (IHBT), Palampur, exhibiting mild mottling and stunting. The causal virus (Cucumber mosaic virus, CMV) was identified and characterized on the basis of host range, aphid transmission, enzyme linked immunosorbent assay (ELISA), DNA-RNA hybridization and reverse transcription polymerase chain reaction (RT-PCR). A complete coat protein (CP) gene was amplified using degenerate primers and sequenced. The CP gene showed nucleotide and amino acid homology up to 97%-98% and 96%-99%, respectively with the sequences of CMV subgroup II. The CP gene also showed homologies of 75%-97% in nucleotide and 77%-96% in amino acid with the CMV Indian isolates infecting various crops. On the basis of sequence homology, it was concluded that CMV-infecting geraniums in India belong to subgroup II.

The study was aimed at evaluating the antioxidant activities of extract of Usnea ghattensis. The antioxidant activity, reducing power, superoxide anion radical scavenging and free radical scavenging activities were studied. The antioxidant activity increased with the increasing amount of extracts (2–20 mg/ml) added to the linoleic acid emulsion. Lipid peroxidation upto 73.3% was inhibited by the extract of 20 mg/ml and 39.2% by α-tocopherol at the same concentration. These effects were statistically significant ( r 2 = 0.876 , P 0.01 ) when compared with control. However, the extract had no significant effect on superoxide anion scavenging by the PMS/NADH-NBT method. Like antioxidant activity, the reducing power and free radical scavenging activity of extract depends on concentration and increasing with increased amount of sample. The reducing power and DPPH radical-scavenging activity of U. ghattensis extract were found greater than the BHA and BHT. The results obtained in the present study indicate that U. ghattensis is a potential source of natural antioxidant.

Various solvent extracts of the lichen Usnea ghattensis showed good antioxidant activity. A methanol extract prevented lipid peroxidation by 87% followed by 65% in Trolox at 20 microg/ml. It also showed superoxide anion scavenging activity and free radical scavenging activity 56% and 73%, respectively. The known antioxidants butylated hydroxytoluene (BHT), butylated hydroxyanisol (BHA) and quercetin at similar concentrations showed superoxide anion scavenging activity of 68, 59 and 47% and free radical scavenging activity 83, 77 and 69%, respectively. In addition, these extracts were inhibitory against Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis and Staphylococcus aureus with MIC values of 5-10 microg/ml.

Prunus necrotic ringspot virus (PNRSV)-free Begonia spp. plants were raised from petioles of virus-infected plants using in vitro techniques. The petioles were grown on MS medium supplemented with 0.2 mg/l NAA and 0.2 mg/l BAP (pH 5.8). For rooting, half-strength MS medium without any plant growth regulators was used. On rooting medium, shoots were subjected to chemotherapy (virazole, 2-thiouracil or 6-azauracil) and thermotherapy (38 °C for 16 h light period and 22 °C for 8 h dark period) separately or in combination. Regenerated plants (treated with chemo- and thermotherapy) were indexed for PNRSV by DAS-ELISA and RT-PCR. An amplified product of 785 bp was obtained by RT-PCR in PNRSV-infected plants. Virazole at a concentration of 20 mg/l was found to be more effective (30 and 20% of PNRSV-free plants as indexed by ELISA and RT-PCR, respectively) in comparison to the other chemicals. Thermotherapy for 25 days gave 35 and 25% PNRSV-free plants as indexed by DAS-ELISA and RT-PCR, respectively. A combination of both treatments gave a good number of PNRSV-free plants (67.5 and 57.5% as indexed by DAS-ELISA and RT-PCR, respectively). At higher concentrations all three chemicals were found to be toxic. Thermotherapy for more than 25 days caused browning of leaves and shoots died.

Background: Diabetes is a metabolic disease prevalent worldwide in all age group of people. The source of diabetes is due to an oxidation process that can produce free radicals. An increase in oxidative free radicals in the body is reported to be one of the several causes of diabetes. The best remedy to combat oxidative stress is the use of antioxidants, which inhibit and scavenge free radicals. Aim: This study has been undertaken to evaluate the antioxidant activity and antidiabetic effect of mulberry leaf extract in diabetic mice. Materials and Methods: Antioxidant activity of mulberry leaves was determined by 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and ferric reducing/antioxidant power (FRAP) assay. Antidiabetic assay of mulberry leaf extract was analyzed by oral administration of leaf extract up to 3 weeks in diabetic mice induced by streptozotocin. Results: In vitro antioxidant activity in both DPPH and FRAP assays showed significantly (P < 0.05) higher inhibition of free radicals than that with ascorbic acid. Diabetic mice fed with mulberry leaf extract showed increment (+25.88%) in body weight and a significant reduction in blood glucose concentration (−71.58%). Further, glucose-6-phosphate dehydrogenase enzyme activity was significantly (P < 0.05) increased, whereas activities of other enzymes particularly catalase, serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase were decreased in diabetic mice after oral administration of mulberry leaf extracts. Histology of liver revealed regeneration of hepatocytes, central vein, and nucleus. Conclusion: This study demonstrated that S-1708 mulberry variety has a potential therapeutic value in diabetes and related complications. SUMMARY Diabetes mellitus is a grave metabolic deviations and responsible for many complications affecting various organs in the human body. In spite of the known antidiabetic medicine available in the market, diabetes and the associated impediments sustained to be a major medical crisis. Medicinal plants have been proven to be useful in diabetes due to their rich therapeutic value. In the current study, S-1708 mulberry variety not only authenticated the earlier results obtained from other medicinal plants but also turn out to be known as a potential source for treating diabetes by demonstrating tremendous ant- diabetic properties. Abbreviations used: S-1708, DPPH, FRAP.

The present study deals with the isolation of mollugin, furomollugin and dehydro-a-lapchone from the chloroform fraction of Rubia cordifolia (Linn.) roots. Mollugin showed inhibition of passive cut...

In the present study, the extract of four natural lichen species Canoparmelia eruptens, Everniastrum cirrhatum, Parmotrema austrosinense and Rimelia cetrata were studied for the source of natural antioxidant and their purified secondary metabolites were evaluated for growth promoting effects on probiotic bacteria Lactobacillus casei. The methanolic fraction of lichen species showed moderate to high antioxidant activity in the order P. austrosinense > E. cirrhatum > C. eruptens > R. cetrata. The lichen metabolites showed antioxidant activity with an IC50 values (μg/ml); lecanoric acid 79–95, salazinic 88–108, atranorin 100–116 and consalazinic acid 119–125. As far as the growth promoting effects of lichen metabolites on L. casei is concerned, lecanoric acid at 100 μg/ml conc. showed high growth stimulating activity in terms of increased dry matter of biomass (56.08 mg) of L. casei. Other lichen metabolites; salazinic acid, atranorin and consalazinic acid produced relatively less dry biomass 43.98 mg, 41.1 mg, 40.68 mg, respectively. However, standard antioxidants butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and Trolox after 36 h produced 39.04–47.81 mg dry biomass. At lower pH the growth promoting activity of lichen metabolites was found stable.

Poly(A) polymerase from wheat embryos was shown to be a phosphoprotein by labelling the enzyme in vivo with [32P]orthophosphate. Excised germinated wheat embryos (48 h) were incubated (12 h) in a sterile solution of sucrose (2%) containing [32P]orthophosphate (25 mCi) for labelling phosphoproteins. Poly(A) polymerase was then purified by ATP-Sepharose affinity chromatography. The ATP-Sepharose fraction showed a single protein-stained band on native PAGE that matched with the activity peak of the unlabelled enzyme. This proved the electrophoretic homogeneity of the enzyme preparation. Serial scanning of the parallel unstained gel showed a single 32P-labelled radioactive peak that matched the protein-stained band. Analysis of 32P-labelled purified poly(A) polymerase on SDS-PAGE also showed a single radioactive band that coincided with the protein-stained band (65 kDa) of a monomeric poly(A) polymerase. Chemical characterization of the 32P-labelled purified poly(A) polymerase by acid hydrolysis showed two radioactive spots by paper chromatography that matched authentic phosphoserine and phosphothreonine. These results suggested that wheat embryo poly(A) polymerase is phosphorylated in vivo at the serine and threonine residues. A significant amount of dephosphorylation (∼ 74%) of poly(A) polymerase was achieved by treating the 32P-labelled purified enzyme with alkaline phosphatase. Addition of alkaline phosphatase along with sodium ortho-vanadate, however, showed a meagre dephosphorylation (∼8%) of 32P-labelled poly(A) polymerase. This is because ortho-vanadate virtually blocked the activity of phosphatase (∼90%) and thus prevented the dephosphorylation of poly(A) polymerase. This enabled us to compare the biological activity of phosphorylated and substantially dephosphorylated forms of poly(A) polymerase. Thus, purified poly(A) polymerase was dephosphorylated with alkaline phosphatase and thereafter, phosphatase activity was blocked in situ by the addition of sodium ortho-vanadate. We observed a substantial decrease (∼ 60%) in poly(A) polymerase activity following treatment with alkaline phosphatase. However, poly(A) polymerase activity was not inhibited to any significant level by alkaline phosphatase in the simultaneous presence of sodium ortho-vanadate. We consider that the partially dephosphorylated form of poly(A) polymerase is less active than the phosphorylated form in wheat embryos.

In the present report, nutritional requirement for the culture of two lichen species Ramalina nervulosa and Ramalina pacifica were studied in order to enhance their growth rate and antioxidant metabolite production. Extract of R. nervulosa cultured in Bold’s basal medium (BBM) showed higher antioxidant activity than R. pacifica cultured in Murashige and Skoog (MS) medium. The lichen species were sub-cultured in standardized nutrient media. R. nervulosa in BBM (1% glucose, 50 ppb asparagines, pH 6.5) yielded 2.76 g biomass with 26.18 mg sekikaic acid, 24.32 mg usnic acid/g dry biomass in a period of 60 days. R. pacifica in MS media (3% sucrose, 100 ppb thiamine, pH 5.9) yielded 3.54 g biomass and 58.92 mg salazinic acid, 40.16 mg usnic acid in the same time period. The standardized culture conditions implemented on bioreactor, R. nervulosa yielded 17.7 g biomass with the production of sekikaic acid 122.8 mg, usnic acid 75.4 mg in 4.5 days. R. pacifica produced 10.3 g biomass along with salazinic acid 200 mg and usnic acid 136.8 mg in the same duration. Lichen secondary metabolites produced in bioreactor showed moderate antioxidant activity; sekikaic acid 42% to 56.4%; salazinic acid 33.6% to 41.9% and usnic acid 19.9% to 29.5%.

Wheat embryo poly(A) polymerase was purified (1348-fold) to electrophoretic homogeneity by adenosine triphosphate(ATP)-Sepharose and concanavalin A(Con A)-agarose affinity chromatography. The purified polymerase was devoid of cryptic nuclease activity after Con A-agarose affinity chromatography. Thus the Con A-agarose fraction showed a linear increase in poly(A) polymerase activity as a function of time up to 90 min. Fractionation of purified enzyme on native PAGE showed a single protein stained band that coincided with the activity peak of poly(A) polymerase. The molecular weight of poly(A) polymerase was 65 000–70 000 as determined by molecular sieve chromatography. A single subunit of purified poly(A) polymerase (mol. wt., 64 000) was observed on SDS-PAGE. This proved the monomeric nature of the enzyme. The binding of poly(A) polymerase to Con A-agarose and its elution with α-methyl mannopyranoside suggested its glycoprotein nature. The apparent Km of poly(A) polymerase for ATP was 86 μM. The 3H-labelled reaction product of purified enzyme binds to oligo(dT)-cellulose affinity matrix. In addition, the putative poly(A) product was not hydrolysed by RNase A and RNase Ti. Thus, it was proved that poly(A) polymerase catalyzes the synthesis of poly(A) sequences.

Antioxidative and hepatoprotective activity of a cultured lichen Usnea ghattensis has been studied. The methanolic extract of cultured lichen U. ghattensis showed good antioxidant activity by preventing lipid peroxidation by 67% and 86% in Trolox-equivalent antioxidant capacity at 20 microg/ml. It also showed superoxide, 1,1-diphenyl-2-picrylhydrazyl, nitric oxide, and hydroxyl radical-scavenging activity, 89%, 89.6%, 94.8%, and 89.6%, respectively, and found levels higher then that known for the synthetic antioxidants butylated hydroxytoluene, butylated hydroxyanisol, and quercetin at 20 microg/ml concentration. The cultured lichen extract also showed hepatoprotection against ethanol-induced toxicity in the mice liver slice culture model by a significant decrease in the antioxidant enzymes, glutathione peroxidase, catalase, and superoxide dismutase, along with a decrease in lipid peroxidation and lactate dehydrogenase release.

Summary The study was aimed to optimize the culture conditions for the production of usnic acid in the cultured cell aggregates composed of symbionts in lichen Usnea ghattensis in vitro. The cultured lichen tissue composed of symbionts appeared after about 2–3 weeks of inoculation in water–agar and malt–yeast extract (MYE) media and shown the production of usnic acid after 2–3 months of inoculation. However, the growth of symbionts was strongly affected by different culture conditions. The addition of excess carbon and nitrogen sources in the media has significantly enhanced the growth as well as usnic acid content. The cultured symbionts in MYE medium having 4% sucrose, 4% polyethyl glycol (PEG) gave 7.63 g dry biomass with 3.9 μg usnic acid/g dry biomass. In water–agar medium having 4% sucrose and 4% PEG gave 3.08 g dry biomass with 1.11 μg usnic acid/g dry biomass. The positive effects of medium on the growth of symbionts and the production of usnic acid are seemed to be due to nutritional factors.

The biologically active compounds, antioxidant activities and free radical scavenging effects of dry matter of cultural medium (DMCM), filtrate (DMF) and different solvent extracts of mycobiont from lichen species Usnea ghattensis in submerged culture (UGSC) were investigated. DMF showed the strongest inhibition of lipid peroxidation as a function of its concentration, and was comparable to the antioxidant activity of BHA at the same concentration of 0.2 mg/mL. The hexane extract of mycobiont had the weakest antioxidant ability, whereas other mycobiont extracts exhibited a modest to strong inhibition of lipid peroxidation and in free radical scavenging activity. The antioxidant activities of mycobiont extracts were correlated with the presence of total polyphenol content in the extract. It was found that antioxidant components in methanol, acetone, ethanol and hexane extracts must be derived from the secondary metabolites produced by the lichen mycobiont. The results presented herein indicate that extracts of cultured mycobiont of lichen Usnea ghattensis could possibly act as chemopreventing agent with respect to free radical-related diseases.

A survey for distribution and abundance of plant parasitic nematodes in fields grown to Lilium in Himachal Pradesh, India at four study sites viz. Nagrota (at 810 m a.s.l.), Palampur (at 1270 m a.s.l.), Sunder Nagar (at 1400 m a.s.l.) and Chail (at 2250 m a.s.l.) was carried out. Moderate (101-500/200 ml soil) to high (501-1000/200 ml soil) populations of phytonematodes including the vectors for plant viruses (Aphelenchoides avenae, Criconemoides spp., Hoplolaimus spp., Longidorus spp., Paratylenchus spp., Pratylenchus spp., Rhabditis spp., Trichodorus spp., Tylenchoryhnchus spp., Tylenchulus spp. and Xiphinema diversicaudatum) were recorded. Mean population of nematodes was positively correlated with pH in all the study sites, negatively correlated with electrical conductivity (EC), percent organic matter (OM%), available potassium (K) and positively correlated with percent carbon (C%), available nitrogen (N) and available phosphorus (P) in all but one study site. The highest incidence of virus-vector nematodes viz. X. diversicaudatum, Longidorus spp. and Trichodorus spp. was recorded at Palampur. Only Strawberry latent ringspot nepovirus (SLRSV) was detected in Lilium cvs. Star Gazer Max and Galeili by Enzyme Linked Immunosorbent Assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) and in X. diversicaudatum associated with the cultivars by RT-PCR. Cucumis sativus used as bait plants showed SLRSV symptoms after 15 days of nematode inoculation and further SLRSV was again detected by ELISA and RT-PCR in C. sativus plants confirming the transmission of SLRSV by X. diversicaudatum in Lilium.