Adhatoda vasica and Calotropis procera as a resource of novel chemical compounds, their biological bioluminescence assay, and investigation of morphological features of bacterial growth through advanced technologies.

This article reports the three principal groups of compounds for the first time from Adhatoda vasica and Calotropis procera plants species using nuclear magnetic resonance methods in which aliphatic, oxy heterocyclic, and tannins compounds were detected from these plants. The leaves of both species were subjected to testing tyrosinase inhibition and antioxidant activities. ATP bioluminescence use for indirect measurement of the amount of organic residue on the surface of the leaves that provide support to microbial growth. The distinguishing characteristics and intraoperative findings of bacterial diseases involved in treatments were conducted against the positive and negative microbial strains using a scanning electron microscope (SEM). The methanolic extracts of leaves of both species were applied to bacterial strains through broth microdilution method to determine the minimum inhabitation concentrations (MICs) for both species. It was concluded that both plants are a rich resource of bioactive compounds. Their extract may also be used to treat various bacterial diseases and in drug manufacturing. HIGHLIGHTS: New chemical compounds of oxy-heterocyclic, aliphatic, and tannins derivatives are isolated from herbal plants as a source of various drugs. 1 H NMR spectrum and 13 C NMR spectrum of each new derivate were calculated. NMR-spectral analysis of new compound of chemistry class was studied and further applied in various bacterial strains. Tyrosinase inhibition property of bacteria strains by application of active compounds on these strains. Agar overlay bioassays were used to evaluate intercellular morphological features of strains applied on extracts by electron microscope (SEM). a-Glucosidase inhibition assay determined with antioxidants activity through FRAP assay methods.

Selenium Alleviates the Adverse Effect of Drought in Oilseed Crops Camelina (Camelina sativa L.) and Canola (Brassica napus L.).

Drought poses a serious threat to oilseed crops by lowering yield and crop failures under prolonged spells. A multi-year field investigation was conducted to enhance the drought tolerance in four genotypes of Camelina and canola by selenium (Se) application. The principal aim of the research was to optimize the crop yield by eliciting the physio-biochemical attributes by alleviating the adverse effects of drought stress. Both crops were cultivated under control (normal irrigation) and drought stress (skipping irrigation at stages i.e., vegetative and reproductive) conditions. Four different treatments of Se viz., seed priming with Se (75 µM), foliar application of Se (7.06 µM), foliar application of Se + Seed priming with Se (7.06 µM and 75 µM, respectively) and control (without Se), were implemented at the vegetative and reproductive stages of both crops. Sodium selenite (Na2SeO3), an inorganic compound was used as Se sources for both seed priming and foliar application. Data regarding physiochemical, antioxidants, and yield components were recorded as response variables at crop maturity. Results indicated that WP, OP, TP, proline, TSS, TFAA, TPr, TS, total chlorophyll contents, osmoprotectant (GB, anthocyanin, TPC, and flavonoids), antioxidants (APX, SOD, POD, and CAT), and yield components (number of branches per plant, thousand seed weight, seed, and biological yields were significantly improved by foliar Se + priming Se in both crops under drought stress. Moreover, this treatment was also helpful in boosting yield attributes under irrigated (non-stress) conditions. Camelina genotypes responded better to Se application as seed priming and foliar spray than canola for both years. It has concluded that Se application (either foliar or priming) can potentially alleviate adverse effects of drought stress in camelina and canola by eliciting various physio-biochemicals attributes under drought stress. Furthermore, Se application was also helpful for crop health under irrigated condition.

Selenium Biofortification: Roles, Mechanisms, Responses and Prospects.

The trace element selenium (Se) is a crucial element for many living organisms, including soil microorganisms, plants and animals, including humans. Generally, in Nature Se is taken up in the living cells of microorganisms, plants, animals and humans in several inorganic forms such as selenate, selenite, elemental Se and selenide. These forms are converted to organic forms by biological process, mostly as the two selenoamino acids selenocysteine (SeCys) and selenomethionine (SeMet). The biological systems of plants, animals and humans can fix these amino acids into Se-containing proteins by a modest replacement of methionine with SeMet. While the form SeCys is usually present in the active site of enzymes, which is essential for catalytic activity. Within human cells, organic forms of Se are significant for the accurate functioning of the immune and reproductive systems, the thyroid and the brain, and to enzyme activity within cells. Humans ingest Se through plant and animal foods rich in the element. The concentration of Se in foodstuffs depends on the presence of available forms of Se in soils and its uptake and accumulation by plants and herbivorous animals. Therefore, improving the availability of Se to plants is, therefore, a potential pathway to overcoming human Se deficiencies. Among these prospective pathways, the Se-biofortification of plants has already been established as a pioneering approach for producing Se-enriched agricultural products. To achieve this desirable aim of Se-biofortification, molecular breeding and genetic engineering in combination with novel agronomic and edaphic management approaches should be combined. This current review summarizes the roles, responses, prospects and mechanisms of Se in human nutrition. It also elaborates how biofortification is a plausible approach to resolving Se-deficiency in humans and other animals.

Enhanced Nicotiana benthamiana immune responses caused by heterologous plant genes from Pinellia ternata.

BACKGROUND: Pinellia ternata is a Chinese traditional medicinal herb, used to cure diseases including insomnia, eclampsia and cervical carcinoma, for hundreds of years. Non-self-recognition in multicellular organisms can initiate the innate immunity to avoid the invasion of pathogens. A design for pathogen independent, heterosis based, fresh resistance can be generated in F1 hybrid was proposed. RESULTS: By library functional screening, we found that P. ternata genes, named as ptHR375 and ptHR941, were identified with the potential to trigger a hypersensitive response in Nicotiana benthamiana. Significant induction of ROS and Callose deposition in N. benthamiana leaves along with activation of pathogenesis-related genes viz.; PR-1a, PR-5, PDF1.2, NPR1, PAL, RBOHB and ERF1 and antioxidant enzymes was observed. After transformation into N. benthamiana, expression of pathogenesis related genes was significantly up-regulated to generate high level of resistance against Phytophthora capsici without affecting the normal seed germination and morphological characters of the transformed N. benthamiana. UPLC-QTOF-MS analysis of ptHR375 transformed N. benthamiana revealed the induction of Oxytetracycline, Cuelure, Allantoin, Diethylstilbestrol and 1,2-Benzisothiazol-3(2H)-one as bioactive compounds. Here we also proved that F1 hybrids, produced by crossing of the ptHR375 and ptHR941 transformed and non-transformed N. benthamiana, show significant high levels of PR-gene expressions and pathogen resistance. CONCLUSIONS: Heterologous plant genes can activate disease resistance in another plant species and furthermore, by generating F1 hybrids, fresh pathogen independent plant immunity can be obtained. It is also concluded that ptHR375 and ptHR941 play their role in SA and JA/ET defense pathways to activate the resistance against invading pathogens.

Antimicrobial investigation of selected soil actinomycetes isolated from unexplored regions of Kashmir Himalayas, India.

The aim of the present study was to isolate and evaluate the antimicrobial potential of soil actinomycetes of Kashmir Himalayas. The secondary metabolites of actinomycetes are the prominent source of antibiotics. A total of 121 morphologically different actinomycete strains were isolated and screened for antimicrobial activity against various human pathogens. The ethyl acetate extract of fermented broth an actinomycete strain, identified as Streptomyces pratensis exhibited significant antimicrobial activity against Staphylococcus aureus ATCC 29213 with MIC 0.25 µg/ml and Mycobacterium tuberculosis Strain H37Rv with MIC 0.062 µg/ml. The strain S. pratensis IIIM06 was grown on large scale and their broth was extracted with ethyl acetate. The extract was subjected to various chromatography techniques which led to the isolation of four compounds whose structures were established as actinomycin C1, actinomycin C2, actinomycin C3 and actiphenol on the basis of spectral data analysis. Actinomycin C1, C2 and C3 exhibited potent antimicrobial activity against S. aureus as well as M. tuberculosis. The isolated indigenous actinomycetes exhibited good antibacterial activity and the study reveals that IIIM06 is a promising strain and could be of great potential for industrial applications.

Contribution of moxifloxacin or levofloxacin in second-line regimens with or without continuation of pyrazinamide in murine tuberculosis.

RATIONALE: High-dose levofloxacin (L) (1,000 mg) was as active as moxifloxacin (M) (400 mg) in an early bactericidal activity trial, suggesting these fluoroquinolones could be used interchangeably. Whether pyrazinamide (Z) contributes sterilizing activity beyond the first 2 months in fluoroquinolone-containing second-line regimens remains unknown. OBJECTIVES: We compared the efficacy of M and high-dose L alone or in combination with ethionamide (Et), amikacin (A), and Z given for 2 or 7 months. METHODS: A pharmacokinetic study was performed to determine the L dose equivalent to 1,000 mg in humans. Treatment started 2 weeks after aerosol infection with Mycobacterium tuberculosis H37Rv. Mice received M or L alone or in combination with 2 months of EtZA followed by 5 months of Et or EtZ. MEASUREMENTS AND MAIN RESULTS: After 2 months of treatment, lung colony-forming unit (CFU) counts were similar in mice receiving either fluoroquinolone alone, but, after 4 and 5 months, CFU counts were 2 log10 lower in mice receiving M. Mice receiving 2MEtZA/3MEt and 2LEtZA/3LEt had 1.0 and 2.7 log10 lung CFUs, respectively. When Z was given throughout, both regimens rendered mice culture negative by 5 months, and most mice did not relapse after 7 months of treatment, with fewer relapses observed in the M group after 6 and 7 months of treatment. CONCLUSIONS: In murine tuberculosis, M had superior efficacy compared with L despite lower serum drug exposures and may remain the fluoroquinolone of choice for second-line regimens. Z contributed substantial sterilizing activity beyond 2 months in fluoroquinolone-containing second-line regimens, largely compensating for L's weaker activity.

An evaluation of aerobic and anaerobic composting of banana peels treated with different inoculums for soil nutrient replenishment.

This study sought to evaluate the efficacy of aerobic and anaerobic composting of inoculated banana peels, and assess the agronomic value of banana peel-based compost. Changes in the chemical composition under aerobic and anaerobic conditions were examined for four formulations of banana peel-based wastes over a period of 12 weeks. The formulations i.e. plain banana peel (B), and a mixture with either cow dung (BC), poultry litter (BP) or earthworm (BE) were separately composted under aerobic and anaerobic conditions under laboratory conditions. Inoculation with either cow dung or poultry litter significantly facilitated mineralization in the order: BP>BC>B. The rate of decomposition was significantly faster under aerobic than in anaerobic composting conditions. The final composts contained high K (>100 g kg(-1)) and TN (>2%), indicating high potential as a source of K and N fertilizer.

Activity of 5-chloro-pyrazinamide in mice infected with Mycobacterium tuberculosis or Mycobacterium bovis.

BACKGROUND & OBJECTIVES: Pyrazinamide is an essential component of first line anti-tuberculosis regimen as well as most of the second line regimens. This drug has a unique sterilizing activity against Mycobacterium tuberculosis. Its unique role in tuberculosis treatment has lead to the search and development of its structural analogues. One such analogue is 5-chloro-pyrazinamide (5-Cl-PZA) that has been tested under in vitro conditions against M. tuberculosis. The present study was designed with an aim to assess the activity of 5-Cl-PZA, alone and in combination with first-line drugs, against murine tuberculosis. METHODS: The minimum inhibitory concentration (MIC) of 5-Cl-PZA in Middlebrook 7H9 broth (neutral pH) and the inhibitory titre of serum from mice that received a 300 mg/kg oral dose of 5-Cl-PZA 30 min before cardiac puncture were determined. To test the tolerability of orally administered 5-Cl-PZA, uninfected mice received doses up to 300 mg/kg for 2 wk. Four weeks after low-dose aerosol infection either with M. tuberculosis or M. bovis, mice were treated 5 days/wk with 5-Cl-PZA, at doses ranging from 37.5 to 150 mg/kg, either alone or in combination with isoniazid and rifampicin. Antimicrobial activity was assessed by colony-forming unit counts in lungs after 4 and 8 wk of treatment. RESULTS: The MIC of 5-Cl-PZA against M. tuberculosis was between 12.5 and 25 µg/ml and the serum inhibitory titre was 1:4. Under the same experimental conditions, the MIC of pyrazinamide was >100 µg/ml and mouse serum had no inhibitory activity after a 300 mg/kg dose; 5-Cl-PZA was well tolerated in uninfected and infected mice up to 300 and 150 mg/kg, respectively. While PZA alone and in combination exhibited its usual antimicrobial activity in mice infected with M. tuberculosis and no activity in mice infected with M. bovis, 5-Cl-PZA exhibited antimicrobial activity neither in mice infected with M. tuberculosis nor in mice infected with M. bovis. INTERPRETATION & CONCLUSION: Our findings showed that 5-Cl-PZA at doses up to 150 mg/kg was not active in chronic murine TB model. Further studies need to be done to understand the mechanism and mode of inactivation in murine model of tuberculosis.

Leeching in the history--a review.

Leeching has enjoyed a good reputation in the past where it had been used in various ailments from fevers to flatulence. The present day scientists have limited its role and the leeching nowadays is being used only in microsurgeries to relieve the venous congestion. This study was designed to explore the possibility of revival of the leech therapy (leeching) which is still being used traditionally as therapeutic agent in various ailments. Leeching is not the outcome of the medieval period but has been in use during the times when there was no concept of the disease and medicament. The earliest clearly documented record of leeches being used for remedial purpose appears in a painting in an Egyptian Tomb of around 1500 BC. The journey of the leech therapy reached its zenith in 17th and 18th century AD in Europe while as during the Arab era the leeches were used medicinally but only for the bloodletting. During the 17th and 18th century AD there was shortage of leeches in certain European countries due to its rigorous use. During early 20th century AD when germ theory was put forward and medical fraternity believed that every disease has its origin from germs and the era of antibacterial therapy gained a pace, the leech therapy was considered the myth of the past. It was in 1970's that the leech therapy was revived by only limited to the microsurgeries to relieve venous congestions. During the 21st century there were certain studies when the leeches were tried in certain ailments like arthritis etc. and the Food and Drug Authority of USA (FDA) gave permission for sale and use of leeches in USA but limited its use in microsurgeries and plastic surgeries only. The aim of this study was to explore different diseases where the role of leech therapy can be seen and clinical trials can be started in this direction. Although the researchers in Regional Research Institute of Unani Medicine, Srinagar, Kashmir, India working under the aegis of Central Council for Research in Unani Medicine started the clinical trials for leech therapy in frost bite way back in 1999, but there are certain unexplored areas where leech therapy can prove beneficial and need is to take up the studies on other disease on larger sample size.

Leachability and phytoavailability of nitrogen, phosphorus, and potassium from different bio-composts under chloride- and sulfate-dominated irrigation water.

Concerns over increased phosphorus (P) application with nitrogen (N)-based compost application have shifted the trend to P-based composed application, but focusing on one or two nutritional elements does not serve the goals of sustainable agriculture. The need to understand the nutrient release and uptake from different composts has been further aggravated by the use of saline irrigation water in the recent scenario of fresh water shortage. Therefore, we evaluated the leachability and phytoavailability of P, N, and K from a sandy loam soil amended with animal, poultry, and sludge composts when applied on a total P-equivalent basis (200 kg ha(-1)) under Cl(-) (NaCl)- and SO4(2-) (Na2SO4)-dominated irrigation water. Our results showed that the concentration of dissolved reactive P (DRP) was higher in leachates under SO(4)(2-) than Cl(-) treatments. Compost amendments differed for DRP leaching in the following pattern: sludge > animal > poultry > control. Maize (Zea mays L.) growth and P uptake were severely suppressed under Cl(-) irrigation compared with SO4(2-) and non-saline treatments. All composts were applied on a total P-equivalent basis, but maximum plant (shoot + root) P uptake was observed under sludge compost amendment (73.4 mg DW(-1)), followed by poultry (39.3 mg DW(-1)), animal (15.0 mg DW(-1)), and control (1.2 mg DW(-1)) treatment. Results of this study reveal that irrigation water dominated by SO4(2-) has greater ability to replace/leach P, other anions (NO3(-)), and cations (K+). Variability in P release from different bio-composts applied on a total P-equivalent basis suggested that P availability is highly dependent on compost source.

Novel chemotherapy for tuberculosis: chemotherapeutic potential of econazole- and moxifloxacin-loaded PLG nanoparticles.

The potential of econazole (ECZ) and moxifloxacin (MOX) individually against tuberculosis (TB) caused by multidrug-resistant and latent Mycobacterium tuberculosis has been demonstrated. In this study, poly-(dl-lactide-co-glycolide) (PLG) nanoparticle-encapsulated ECZ and MOX were evaluated against murine TB (drug susceptible) in order to develop a more potent regimen for TB. PLG nanoparticles were prepared by the multiple emulsion and solvent evaporation technique and were administered orally to mice. A single oral dose of PLG nanoparticles resulted in therapeutic drug concentrations in plasma for up to 5 days (ECZ) or 4 days (MOX), whilst in the organs (lungs, liver and spleen) it was up to 6 days. In comparison, free drugs were cleared from the same organs within 12-24h. In M. tuberculosis-infected mice, eight oral doses of the formulation administered weekly were found to be equipotent to 56 doses (MOX administered daily) or 112 doses (ECZ administered twice daily) of free drugs. Furthermore, the combination of MOX+ECZ proved to be significantly efficacious compared with individual drugs. Addition of rifampicin (RIF) to this combination resulted in total bacterial clearance from the organs of mice in 8 weeks. PLG nanoparticles appear to have the potential for intermittent therapy of TB, and combination of MOX, ECZ and RIF is the most potent.

Chemotherapeutic evaluation of alginate nanoparticle-encapsulated azole antifungal and antitubercular drugs against murine tuberculosis.

The present study was designed to evaluate the chemotherapeutic potential of alginate nanoparticle-encapsulated econazole and antitubercular drugs (ATDs) against murine tuberculosis. Alginate nanoparticles encapsulating econazole and ATDs were prepared by the cation-induced controlled gelification of alginate and were characterized. Drugs were analyzed by high-performance liquid chromatography. All the ATDs were detected above minimum inhibitory concentrations for as long as 15 days and econazole until the day 8 in organs (lungs, liver, and spleen) after administration of encapsulated drugs, whereas free drugs remained detectable for only 12 to 24 hours. Eight doses of alginate nanoparticle-encapsulated econazole or 112 doses of free econazole reduced bacterial burden by more than 90% in the lungs and spleen of mice infected with Mycobacterium tuberculosis. Econazole (free or encapsulated) could replace rifampicin and isoniazid during chemotherapy of murine tuberculosis. Alginate nanoparticles reduced the dosing frequency of azoles and ATDs by 15-fold. Alginate nanoparticles are the ideal carriers of azole and antitubercular drugs, which can reduce dosing frequency of azoles as well as ATDs for the better management of tuberculosis.

The potential of azole antifungals against latent/persistent tuberculosis.

The aim of the present study was to evaluate the chemotherapeutic potential of econazole against latent tuberculosis. The activity of econazole and clotrimazole was tested against the latent bacilli (Mycobacterium tuberculosis H(37)Rv) developed by nutrient starvation under in vitro conditions and by drugs under in vivo conditions. The latent bacteria developed under in vitro latent conditions were acid-fast negative, nonreplicating, resistant to conventional antitubercular drugs and showed low respiration rates. Econazole as well as clotrimazole were found to have strong antimycobacterial potential against latent Mycobacterium tuberculosis under in vitro conditions as seen by reductions in colony-forming units. Further, econazole prevented the formation of drug-induced latency and significantly reduced bacterial burden from lungs and spleens of latent tuberculosis-infected mice. We conclude that azole drugs bear significant therapeutic potential against latent tuberculosis.

Inhalable alginate nanoparticles as antitubercular drug carriers against experimental tuberculosis.

Pharmacokinetic and chemotherapeutic studies have been carried out with aerosolised alginate nanoparticles encapsulating isoniazid (INH), rifampicin (RIF) and pyrazinamide (PZA). The nanoparticles were prepared by cation-induced gelification of alginate and were 235.5 +/- 0 nm in size, with drug encapsulation efficiencies of 70-90% for INH and PZA and 80-90% for RIF. The majority of particles (80.5%) were in the respirable range, with mass median aerodynamic diameter of 1.1 +/- 0.4 microm and geometric standard deviation of 1.71 +/- 0.1 microm. The relative bioavailabilities of all drugs encapsulated in alginate nanoparticles were significantly higher compared with oral free drugs. All drugs were detected in organs (lungs, liver and spleen) above the minimum inhibitory concentration until 15 days post nebulisation, whilst free drugs stayed up to day 1. The chemotherapeutic efficacy of three doses of drug-loaded alginate nanoparticles nebulised 15 days apart was comparable with 45 daily doses of oral free drugs. Thus, inhalable alginate nanoparticles can serve as an ideal carrier for the controlled release of antitubercular drugs.