Facile synthesis and nanoscale features of a nanostructured nordihydroguaiaretic acid analog for therapeutic applications.

BACKGROUND: Nordihydroguaiaretic acid (NDGA) is a plant lignan obtained from creosote bush, known to possess anti-oxidant, anti-cancer and anti-viral activities and is being used in traditional medicine. However, toxicity studies indicated liver and kidney damage despite its immense medicinal properties. There has been a recent increase of curiosity in the chemical synthesis of NDGA derivatives for therapeutic applications. NDGA derivatives have been developed as better alternatives to NDGA and for targeted delivery to the site of tissue by chemical derivatives. In this regard, an analog of NDGA, Acetyl NDGA (Ac-NDGA), has been synthesized based on a previous procedure and formulated as a nanostructured complex with Polycaprolactone/Polyethylene glycol polymer matrices, by o/w solvent evaporation method. RESULTS: The drug-incorporated polymeric nanospheres exhibited a drug load of 10.0 ± 0.5 µg drug per mg of nanospheres in acetonitrile solvent with 49.95 ± 10% encapsulation efficiency and 33-41% drug loading capacity with different batches of nanospheres preparation. The in vitro drug release characteristics indicated 82 ± 0.25% drug release at 6 h in methanol. Further, the nanospheres have been characterized extensively to evaluate their suitability for therapeutic delivery. CONCLUSIONS: The present studies indicate a new and efficient formulation of the nanostructured AcNDGA with good therapeutic potential.

Nordihydroguaiaretic Acid in Therapeutics: Beneficial to Toxicity Profiles and the Search for its Analogs.

Nordihydroguaiaretic acid (NDGA) is a plant lignan obtained from creosote bush, Larrea tridentata and is known to possess antioxidant, anticancer activities and is used in traditional medicine in North America and Mexico. However, its prolonged consumption leads to liver damage and kidney dysfunction. Despite its toxicity and side effects, there is little awareness to forbid its consumption and its use in the treatment of medical ailments has continued over the years. Several reports discuss its therapeutic efficiency and its medical applications have tremendously been on the rise to date. There has been a recent surge of interest in the chemical synthesis of NDGA derivatives for therapeutic applications. NDGA derivatives have been developed as better alternatives to NDGA. Although several NDGA derivatives have been chemically synthesized as evidenced by recent literature, there is a paucity of information on their therapeutic efficacies. This review is to highlight the medicinal applications of NDGA, its toxicity evaluations and discuss the chemical derivatives of NDGA synthesized and studied so far and suggest to continue research interests in the development of NDGA analogs for therapeutic applications. We suggest that NDGA derivatives should be investigated more in terms of chemical synthesis with preferred conformational structures and exploit their biological potentials with future insights to explore in this direction to design and develop structurally modified NDGA derivatives for potential pharmacological properties.

Study on Incentives for Glaucoma Medication Adherence (SIGMA): study protocol for a randomized controlled trial to increase glaucoma medication adherence using value pricing.

BACKGROUND: Many glaucoma patients do not adhere to their medication regimens because they fail to internalize the (health) costs of non-adherence, which may not occur until years or decades later. Behavioural economic theory suggests that adherence rates can be improved by offering patients a near-term benefit. Our proposed strategy is to offer adherence-contingent rebates on medication and check-up costs. This form of value pricing (VP) ensures that rebates are granted only to those most likely to benefit. Moreover, by leveraging loss aversion, rebates are expected to generate a stronger behavioural response than equivalent financial rewards. METHODS/DESIGN: The main objective of the Study on Incentives for Glaucoma Medication Adherence (SIGMA) is to test the VP approach relative to usual care (UC) in improving medication adherence. SIGMA is a randomized, controlled, open-label, single-centre superiority trial with two parallel arms. A total of 100 non-adherent (Morisky Medication Adherence Scale ≤6) glaucoma patients from the Singapore National Eye Centre are block-randomized (blocking factor: single versus multiple medications users) into the VP and UC arms in a 1:1 ratio. The treatment received by VP patients will be strictly identical to that received by UC patients, with the only exception being that VP patients can earn either a 50 % or 25 % rebate on their glaucoma-related healthcare costs conditional on being adherent on at least 90 % or 75 % of days as measured by a medication event monitoring system. Masking the arm allocation will be precluded by the behavioural nature of the intervention but blocking size will not be disclosed to protect concealment. The primary outcome is the mean change from baseline in percentage of adherent days at month 6. A day will be counted as adherent when the patients take all their medication(s) within the appropriate dosing windows. DISCUSSION: This trial will provide evidence on whether adherence-contingent rebates can improve medication adherence among non-adherent glaucoma patients, and more generally whether this approach represents a promising strategy to cost-effectively improve chronic disease management. TRIAL REGISTRATION: NCT02271269 . Registered on 19 October 2014.

Design and baseline characteristics of participants in the TRial of Economic Incentives to Promote Physical Activity (TRIPPA): a randomized controlled trial of a six month pedometer program with financial incentives.

Non-communicable diseases (NCDs) are emerging as the predominant global health challenge of this century. Physical inactivity is one of the primary risk factors for NCDs. Therefore, increasing physical activity levels is a public health imperative. The arrival of affordable wearable technologies, such as wireless pedometers, provides one strategy for encouraging walking. However, the effectiveness of these technologies in promoting sustained behavior change has not been established. Insights from economics suggest that incentives may be a useful strategy for increasing maintenance and effectiveness of behavior change interventions, including physical activity interventions that rely on wearable technologies. The aim of this trial is to test the effectiveness of a common wireless pedometer with or without one of two types of incentives (cash or donations to charity) for reaching weekly physical activity goals. We present here the design and baseline characteristics of participants of this four arm randomized controlled trial. 800 full-time employees (desk-bound office workers) belonging to 15 different worksites (on average, 53 (sd: 37) employees at each worksite) were successfully randomized to one of four study arms. If shown to be effective, wearable technologies in concert with financial incentives may provide a scalable and affordable health promotion strategy for governments and employers seeking to increase the physical activity levels of their constituents.

Inducible chromate reductase exhibiting extracellular activity in Bacillus methylotrophicus for chromium bioremediation.

Chromium toxicity is one of the major causes of environmental pollution due to its heavy discharge in industrial wastewaters. Chromate reduction is a viable method to detoxify hexavalent chromium to nontoxic trivalent species mediated by enzymes and metabolites. A new Bacillus methylotrophicus strain was isolated from tannery sludge and was an efficient candidate for chromate reduction. An initial chromate reductase activity of 212.84 U/mg protein was obtained at 48 h in a low-cost defined medium formulation with 0.25 mM chromate. The extracellular enzyme was inducible at 12h substrate addition with 312.99 U/mg at 48 h. Reduced glutathione was required for enhanced specific activity of 356.48 U/mg. Enzyme activity was optimum at pH 7.0 and at 30 °C, and was stable in the presence of EDTA, DTT and metal ions. The enzyme exhibited a Vmax of 59.89 µM/min/mg protein and a Km of 86.5 µM, suggesting feasibility of the reaction with K2Cr2O7 as substrate. Application of the crude reductase in tannery effluent resulted in 91.3% chromate reduction at 48 h. An enzyme-mediated chromate reduction process has therefore been developed for bioremediation of toxic chromium sp. in industrial effluents.

Facile production of ZnS quantum dot nanoparticles by Saccharomyces cerevisiae MTCC 2918.

Microbial synthesis of nanoparticles is a green route towards ecofriendly measures to overcome the toxicity and non-applicability of nanomaterials in clinical uses obtained by conventional physical and chemical approaches. Nanoparticles in the quantum regime have remarkable characteristics with excellent applicability in bioimaging. Yeasts have been commercially exploited for several industrial applications. ZnS nanoparticles as semiconductor quantum dots have mostly been synthesized by bacterial species. Here in, we have attempted to produce ZnS nanoparticles in quantum regime by Saccharomyces cerevisiae MTCC 2918 fungus and characterize its size and spectroscopic properties. Intracellular ZnS nanoparticles were produced by a facile procedure and freeze thaw extraction using 1mM zinc sulfate. The ZnS nanoparticles showed surface plasmon resonance band at 302.57nm. The ZnS nanoparticles were in low yield and in the size range of 30-40nm. Powder XRD analysis revealed that the nanoparticles were in the sphalerite phase. Photoluminescence spectra excited at 280nm and 325nm revealed quantum confinement effects. This suggests that yeasts have inherent sulfate metabolizing systems and are capable fungal sources to assimilate sulfate. Further insights are required to identify the transport/reducing processes that may have caused the synthesis of ZnS nanoparticles such as an oxidoreductase enzyme-mediated mechanism.

Osmotic shock augments ethanol stress in Saccharomyces cerevisiae MTCC 2918.

Yeast cells sense and respond to hypertonicity. Saccharomyces cerevisiae MTCC 2918 was tested for its metabolic status in 1 M NaCl by cell viability analysis, intracellular glycerol content and total antioxidant capacity. Yeast cell viability was maximum in 1 M NaCl and 24 h addition of 1 M NaCl was effective in induction of hyperosmolarity. Increased glycerol contents in cells treated with salt indicated adaptation to osmotic stress with a maximum of 240.87 ± 0.38 mg/g dry weight (DW) at 72 h. The total antioxidant status with 1 M NaCl was 9.29 ± 0.39 mM/g DW at 96 h reflecting free radical quenching to overcome stress with increasing growth period. Considering that pre-adaptation to one type of stress evoked a protective response to other stress factors, we have attempted the cross adaptation of osmotic shock to high ethanol concentrations. In effect, we observed that osmotic shock lowered the cell survival by augmentation of cell toxicity by ethanol due to stress induction during exponential phase. Glycerol accumulation to an order of 470.27 ± 0.53 mg/g DW at 48 h in 1 M NaCl and 12% ethanol indicated that both stresses culminated in membrane disruption further leading to cell burst and contributed to the stress overload.

Molecular cloning of OSP94: A significant biomarker protein of hypertensive human heart and a member of HSP110 family.

Heat shock proteins (HSPs) are upregulated in response to stress and play a protective function in refolding of cellular proteins. In hypertension, the heart is a vital organ that requires examination and investigation, and primary induction of HSPs is predominantly effected. Hypertension results from osmotic imbalance during renin-angiotensin cycle inefficiency. Osmotic stress protein 94 (OSP94) is a stress protein induced upon osmotic imbalance. It is therefore necessary to analyze its precise role in the hypertensive heart. We have first reported the cloning and expression of human heart OSP94 followed by an analysis of gene sequence and protein homology. Directional cloning of OSP94 by PCR amplification yielded a 2.5 kb amplicon and was cloned into pET-15b. Site-directed mutagenesis was essentially followed. mRNA expression levels were evaluated in correlation with HSPs. Gene analysis indicated a 2520 bp sequence with an 838-amino acid protein complement. Protein homology revealed highly conserved sequence similarity among mammalian sequences. Structural predictions of OSP94 protein were also investigated. OSP94 is therefore recognized as a significant stress protein for investigations in hypertensive heart tissues and is a highly conserved protein in the HSP110 subfamily.

Understanding structural features of microbial lipases--an overview.

The structural elucidations of microbial lipases have been of prime interest since the 1980s. Knowledge of structural features plays an important role in designing and engineering lipases for specific purposes. Significant structural data have been presented for few microbial lipases, while, there is still a structure-deficit, that is, most lipase structures are yet to be resolved. A search for 'lipase structure' in the RCSB Protein Data Bank (http://www.rcsb.org/pdb/) returns only 93 hits (as of September 2007) and, the NCBI database (http://www.ncbi.nlm.nih.gov) reports 89 lipase structures as compared to 14719 core nucleotide records. It is therefore worthwhile to consider investigations on the structural analysis of microbial lipases. This review is intended to provide a collection of resources on the instrumental, chemical and bioinformatics approaches for structure analyses. X-ray crystallography is a versatile tool for the structural biochemists and is been exploited till today. The chemical methods of recent interests include molecular modeling and combinatorial designs. Bioinformatics has surged striking interests in protein structural analysis with the advent of innumerable tools. Furthermore, a literature platform of the structural elucidations so far investigated has been presented with detailed descriptions as applicable to microbial lipases. A case study of Candida rugosa lipase (CRL) has also been discussed which highlights important structural features also common to most lipases. A general profile of lipase has been vividly described with an overview of lipase research reviewed in the past.

Mixed substrate solid state fermentation for production and extraction of lipase from Aspergillus niger MTCC 2594.

A novel mixed substrate solid-state fermentation (SSF) process has been developed for Aspergillus niger MTCC 2594 using wheat bran (WB) and gingelly oil cake (GOC) and the results showed that addition of GOC to WB (WB : GOC, 3 : 1, w/w) increased the lipase activity by 36.0% and the activity was 384.3+/-4.5 U/g dry substrate at 30 degrees C and 72 h. Scale up of lipase production to 100 g and 1 kg tray-level batch fermentation resulted in 95.0% and 84.0% of enzyme activities respectively at 72 h. A three-stage multiple contact counter-current extraction yielded 97% enzyme recovery with a contact time of 60 min. However, extraction by simple percolation and plug-flow methods resulted in decreased enzyme recoveries. The mixed substrate SSF process has resulted in a significant increase in specific activity (58.9%) when compared to a submerged fermentation (SmF) system. Furthermore, an efficient process of extraction has been standardized with this process. Use of GOC along with WB as potential raw materials for enzyme production could be of great commercial significance. This is the first report on the production and extraction of lipase from Aspergillus niger using mixed solid substrates, WB and GOC, which are potential raw materials for the production of enzymes and other value-added products.

Bioaccumulation and biosorption of chromium by Aspergillus niger MTCC 2594.

Chromium toxicity is of prime concern due to chrome tanning processes in the leather sector. Chrome tanning results in the discharge of toxic levels of chromium causing pollution hazards. Chromium levels of Cr(III) and Cr(VI) were high above permissible limits in chrome samples after chrome tanning. The potential of Aspergillus niger MTCC 2594 to accumulate chromium as well as its biosorption capacity is investigated in this study. Bioaccumulation of Cr(III) and Cr(VI) in the spent chrome liquor has resulted in a 75-78% reduction of the initial Cr content in 24-36 h. A. niger biomass is found to be very effective in the biosorption of Cr(III) and Cr(VI) in spent chrome liquor. Maximum adsorption of 83% for biosorption of Cr(III) at 48 h and 79% of Cr(VI) at 36 h in spent chrome liquor is observed. The biosorption characteristics fit well with Langmuir and Freundlich isotherms and the adsorption parameters are evaluated. The biosorption of Cr also follows Lagergren kinetics. A. niger biomass is effectively used for the biosorption of chromium with 79-83% Cr removal in 36-48 h.

Strain improvement of Aspergillus niger for enhanced lipase production.

The enhancement of lipase production from Aspergillus niger was attempted by ultraviolet (UV) and nitrous acid mutagenesis, and the mutants were selected on media containing bile salts. Nitrous acid mutants exhibited increased efficiency for lipase production when compared with UV mutants in submerged fermentation. The hyperproducing UV and nitrous acid mutants were further subjected to a second step of mutagenesis to devise an economical and ecofriendly technique for lipase production by the effective use of hydrocarbons. One percent kerosene was found to be optimal for lipase production, and one of the mutant strains NAII exhibited 2.53 times more increased lipase activity than the parental strain did. This investigation indicates a possible role for the A. niger mutant strains in the biodegradation of oil-polluted environments for the development of ecofriendly technologies.