Shikonin and Alkannin inhibit ATP synthase and impede the cell growth in Escherichia coli.

Naturally occurring naphthoquinones, shikonin and alkannin, are important ingredients of traditional Chinese medicine Zicao. These constituents are reported to have many therapeutic uses, such as wound healing; scar treatment; and anti-inflammation, anti-acne, anti-ulcer, anti-HIV, anticancer, and antibacterial properties. The primary objective of this investigation was to explore the effect of shikonin and alkannin on Escherichia coli ATP synthase and its cell growth. Shikonin caused complete (100 %) inhibition, and alkannin caused partial (79 %) inhibition of wild-type E. coli ATP synthase. Both caused partial (4 %-27 %) inhibition of ATP synthase with genetically modified phytochemical binding site. The growth inhibition of strains expressing normal, deficient, and mutant ATP synthase by shikonin and alkannin, corroborated the inhibition observed in isolated normal wild-type and mutant ATP synthase. Trivial inhibition of mutant enzymes indicated αR283D, αE284R, ßV265Q, and γT273A are essential for formation of the phytochemical binding site where shikonin and alkannin bind. Further, shikonin was a potent inhibitor of ATP synthase than alkannin. The antimicrobial properties of shikonin and alkannin were tied to the binding at phytochemical site of microbial ATP synthase. Selective targeting of bacterial ATP synthase by shikonin and alkannin may be an advantageous alternative to address the antibiotic resistance issue.

Probing the physicochemical characteristics of carrot sauce during storage.

Globally, the prevalence of vit-A deficiency disorders i.e., xerophthalmia and nyctalopia is increasing especially in teenagers due to lifestyle shifts and undernutrition. This research was designed to develop carrot-supplemented tomato sauce to overcome vit-A deficiency and its related disorders. The carrot sauce was formulated with the addition of 50, 60, and 70% carrot pulp in tomato paste. The prepared sauce samples were tested for physical and biochemical changes in beta carotene (BC), lycopene, viscosity, pH, total soluble solids, titratable acidity, total plate count, and sensory parameters for 12 weeks. A non-significant effect of storage on BC, lycopene, and total soluble solids was observed. The total plate count, acidity, pH, and viscosity were influenced significantly. Sauce containing 60% of the carrot paste showed good sensory characteristics and 42.39 µg/g BC for the whole period of storage. It is concluded that carrot sauce can be used as tomato ketchup replacers to boost the overall quality of life by fighting against vit-A deficiency disorders.

Nutritional characterization of the extrusion-processed micronutrient-fortified corn snacks enriched with protein and dietary fiber.

The current study focused on developing protein- and dietary fiber-enriched, micronutrient-fortified corn snacks using extrusion technology. Corn, soybean, and chickpea flour were used to develop micronutrient-fortified (Fe, Zn, I, and vitamin A, and C) extruded snacks, followed by an exploration of their nutritional traits. Soybean and chickpea were supplemented discretely (20-40/100 g) or in a combination of both (10:10, 15:15, and 20:20/100 g). According to the results, the relative proportion of the raw material composition was reflected in corn snacks' proximate composition and mineral and vitamin levels. Corn snacks with 40/100 g soy flour showed the best nutrient profile, with a maximum percent increase in protein (171.9%) and fiber (106%), as compared to the snacks developed using chickpea and/or mixed supplementation with soy and chickpea. Total dietary fiber (18.44 ± 0.34%), soluble dietary fiber (10.65 ± 0.13%), and insoluble dietary fiber (7.76 ± 0.38%) were also found to be highest in the soy-supplemented snacks (40/100 g). It was discovered that 100 g of corn snacks could provide 115-127% of the RDA for iron, 77-82% of the RDA for zinc, 90-100% of the RDA for vitamin A, and 45-50% of the RDA for vitamin C. The results for the effect of extrusion processing on amino acids showed a 2.55-45.1% reduction in essential amino acids, with cysteine and valine showing the greatest decrease and leucine and tryptophan remaining relatively stable during extrusion.

Production, functional stability, and effect of rhamnolipid biosurfactant from Klebsiella sp. on phenanthrene degradation in various medium systems.

The present study investigated the stability and efficacy of a biosurfactant produced by Klebsiella sp. KOD36 under extreme conditions and its potential for enhancing the solubilization and degradation of phenanthrene in various environmental matrices. Klebsiella sp. KOD36 produced a mono-rhamnolipids biosurfactant with a low critical micelle concentration (CMC) value. The biosurfactant was stable under extreme conditions (60 °C, pH 10 and 10% salinity) and could lower surface tension by 30% and maintained an emulsification index of > 40%. The emulsion index was also higher (17-43%) in the presence of petroleum hydrocarbons compared to synthetic surfactant Triton X-100. Investigation on phenanthrene degradation in three different environmental matrices (aqueous, soil-slurry and soil) confirmed that the biosurfactant enhanced the solubilization and biodegradation of phenanthrene in all matrices. The high functional stability and performance of the biosurfactant under extreme conditions on phenanthrene degradation show the great potential of the biosurfactant for remediation applications under harsh environmental conditions.

α-Glucosidase Inhibitory, Anti-Oxidant, and Anti-Hyperglycemic Effects of Euphorbia nivulia-Ham. in STZ-Induced Diabetic Rats.

In this study, we aimed to investigate the antidiabetic effects of Euphorbia nivulia (En), native to Cholistan Desert area of Bahawalpur, Pakistan. First, we performed high-performance liquid chromatography analysis and found that this plant contains ferulic acid, gallic acid, quercetin, benzoic acid, polyphenols, and flavonoids. Then, we performed in vitro and in vivo studies to assess its effects on diabetic Wistar rat model. The experiments were performed and compared with control drug glibenclamide. The 70% hydroalcoholic extract of En exhibited 97.8% in vitro α-glucosidase inhibitory effect at a dose of 1.0 mg/mL. We orally administered the extract of En and control drug to the streptozotocin (STZ)-induced diabetic rats and analyzed its antidiabetic effects. We found that the extract of En with a dose of 500 mg/kg/body weight exhibited significant effect to reduce blood glucose in STZ-induced rats as compared with the control group (P < .001). Our histological data also showed that the extract significantly improved the histopathology of pancreas. Collectively, both in vitro and in vivo studies revealed that En possesses α-glucosidase inhibitory, antioxidant, and anti-hyperglycemic effect in STZ-induced diabetic rats.

Application of Rice Grain Husk Derived Biochar in Ameliorating Toxicity Impacts of Cu and Zn on Growth, Physiology and Enzymatic Functioning of Wheat Seedlings.

Livestock and poultry manure containing high levels of copper and zinc have led to contamination of farmland and products which could have an impact on human health. Biochar is an inexpensive and efficient heavy metal absorbent. In the present study, we have used biochar to mitigate the effects of heavy metals on the growth of wheat seedlings. The study showed that the effects of heavy metals on wheat seedlings growth were mitigated by increasing exposure to biochar. Compared to the control group, the germination potential, germination rate, germination index and vigor index of wheat seedlings with supplemented biochar increased significantly. Moreover, the specific activity of catalase, peroxidase, superoxide dismutase decreased and chlorophyll contents increased, which promote wheat growth and suggests that the addition of biochar could reduce the effects of heavy metals on wheat seedlings.

Does posttreatment thymoquinone reverse high-dose atorvastatin-induced hepatic oxidative injury in rats?

Atorvastatin (ATO) was commonly used to lower blood cholesterol, but it caused harmful effects to organs, including the liver. Thymoquinone (TQ), a prominent constituent of Nigella sativa, has antioxidant, antiinflammatory, antiapoptotic, antimicrobial, and anticancer activity. The current study investigated the mechanism of ATO-induced hepatotoxicity, whether posttreatment TQ could reverse ATO-induced hepatic injury, and the mechanism of action of TQ as a hepatoprotective agent. Forty adult male Sprague Dawley rats were divided into four equal groups: control, TQ-treated, ATO-treated, and combined ATO/TQ-treated. Rats were treated for 8 weeks and 10 days and euthanized by cervical dislocation 3 days after the last treatment. Blood samples and livers were tested for liver enzymes, oxidative stress, and apoptosis markers and used for histopathological and ultrastructural examination. The ATO-treated group showed an increase in liver enzymes, decreases in reduced glutathione and catalase, and increases in the malondialdehyde lipid peroxidation marker, protein carbonylation, and caspase 3 activity. Posttreatment TQ in the ATO/TQ-treated group seemed to reverse these changes. Histopathological and ultrastructural examination supported these data. Results from the current study suggested that posttreatment TQ may reverse oxidative stress injury in rat liver produced by ATO, suggesting a potential clinical application of using TQ to prevent ATO-induced hepatic injury.

A Therapeutic Connection between Dietary Phytochemicals and ATP Synthase.

For centuries, phytochemicals have been used to prevent and cure multiple health ailments. Phytochemicals have been reported to have antioxidant, antidiabetic, antitussive, antiparasitic, anticancer, and antimicrobial properties. Generally, the therapeutic use of phytochemicals is based on tradition or word of mouth with few evidence-based studies. Moreover, molecular level interactions or molecular targets for the majority of phytochemicals are unknown. In recent years, antibiotic resistance by microbes has become a major healthcare concern. As such, the use of phytochemicals with antimicrobial properties has become pertinent. Natural compounds from plants, vegetables, herbs, and spices with strong antimicrobial properties present an excellent opportunity for preventing and combating antibiotic resistant microbial infections. ATP synthase is the fundamental means of cellular energy. Inhibition of ATP synthase may deprive cells of required energy leading to cell death, and a variety of dietary phytochemicals are known to inhibit ATP synthase. Structural modifications of phytochemicals have been shown to increase the inhibitory potency and extent of inhibition. Sitedirected mutagenic analysis has elucidated the binding site(s) for some phytochemicals on ATP synthase. Amino acid variations in and around the phytochemical binding sites can result in selective binding and inhibition of microbial ATP synthase. In this review, the therapeutic connection between dietary phytochemicals and ATP synthase is summarized based on the inhibition of ATP synthase by dietary phytochemicals. Research suggests selective targeting of ATP synthase is a valuable alternative molecular level approach to combat antibiotic resistant microbial infections.

Does silybin protect against toxicity induced by polymyxin E in rat kidney?

AIMS: Although once a common antibiotic, polymyxin E fell out of favor after reports of its nephrotoxicity. However, recent concerns with gram-negative bacteria, which are resistant to multiple antibiotics, have resulted in increased interest in polymyxin E. Silybin is a known antihepatotoxic drug and may have potential for protecting the kidney from polymyxin E. Therefore, the aim of the current study was to evaluate whether silybin affected the damages produced by polymyxin E on the rat kidney. METHODS: Four groups of rats with 10 rats per group were included in the study: control (no treatment, group I), vehicle (control vehicle treatment, group II), polymyxin E treatment (group III), and polymyxin E and silybin treatment (group IV). Groups II-IV received intravenous treatment twice a day for 7 days. All rats were euthanized after 7 days. Histological, ultrastructural, and morphometric analyses were performed on the rats' kidney tissues. RESULTS: Analysis of tissues from group III showed differences from groups I and II, such as glomerular and tubular affection and changes in morphometric measures. Results for group IV were more similar to those of groups I and II than those of group III. CONCLUSIONS: Our results suggested that administering silybin with polymyxin E alleviated polymyxin E-induced nephrotoxicity in the rat kidney. Future biochemical studies should investigate whether silybin could ameliorate the nephrotoxicity caused by polymyxin E in rats and whether concomitant administration of silybin could be an effective clinical pharmacological strategy to protect against polymyxin E-induced insult in human kidneys.

Significance of αThr-349 in the catalytic sites of Escherichia coli ATP synthase.

This paper describes the role of α-subunit VISIT-DG sequence residue αThr-349 in the catalytic sites of Escherichia coli F1Fo ATP synthase. X-ray structures show the highly conserved αThr-349 in the proximity (2.68 Å) of the conserved phosphate binding residue ßR182 in the phosphate binding subdomain. αT349A, -D, -Q, and -R mutations caused 90-100-fold losses of oxidative phosphorylation and reduced ATPase activity of F1Fo in membranes. Double mutation αT349R/ßR182A was able to partially compensate for the absence of known phosphate binding residue ßR182. Azide, fluoroaluminate, and fluoroscandium caused insignificant inhibition of αT349A, -D, and -Q mutants, slight inhibition of the αT349R mutant, partial inhibition of the αT349R/ßR182A double mutant, and complete inhibition of the wild type. Whereas NBD-Cl (7-chloro-4-nitrobenzo-2-oxa-1,3-diazole) inhibited wild-type ATPase and its αT349A, -D, -R, and -Q mutants essentially completely, ßR182A ATPase and double mutant αT349A/ßR182A were inhibited partially. Inhibition characteristics supported the conclusion that NBD-Cl reacts in ßE (empty) catalytic sites, as shown previously by X-ray structure analysis. Phosphate protected against NBD-Cl inhibition in the wild type, αT349R, and double mutant αT349R/ßR182A but not in αT349A, αT349D, or αT349Q. The results demonstrate that αThr-349 is a supplementary residue involved in phosphate binding and transition state stabilization in ATP synthase catalytic sites through its interaction with ßR182.

Highlighting the implications of selenium dispersion from disposal of Kahota Industrial Triangle area, Islamabad, Pakistan using three-dimension solute transport model.

This study highlights the implications of selenium (Se) dispersion in groundwater flow regimes of Kahota Industrial Triangle area located adjacent to the Soan River, Islamabad. Initially, a regional groundwater 3-D flow model has been developed, calibrated to the known observed heads of 24 water wells, verified, and confirmed that convergence has actually arrived to satisfy the steady state condition. Later, the transient simulation was carried out adding in the known recharge, storage factor, porosity, and observed drawdown matched with the simulated drawdown that appears to fall in close agreement with a difference of 0.25 m. As such the steady state groundwater model has facilitated to understand the mechanism of groundwater flow regimes in reference to the implications of selenium dispersion from disposal of Kahota Industrial Triangle area. Thirty-five water samples were collected mainly from the industrial water wells for the evaluation of heavy metals. Selenium being the major contributor of pollution has been short listed to monitor its dispersion using a solute transport model modular three-dimensional transport model (MT3D). Chemical parameters related to selenium characteristics including horizontal and vertical transverse dispersivity/longitudinal dispersivity, effective molecular diffusion coefficient and bulk density of the porous medium of aquifers have been used in MT3D contaminant transport model. MT3D is run for 30 years in steady state condition. As usual first run did not produce the exact field conditions. Therefore, the contaminant transport model is calibrated against the 32 values of observed selenium concentrations in boreholes by minor adjustments in the chemical parameter values. The final calibration has been achieved with residual value of 3.88x10(-5) Kg/m3. Seven hypothetical observation wells are used to monitor the selenium concentrations over a long-term period of time.

Determination of toxic inorganic elements pollution in ground waters of Kahuta Industrial Triangle Islamabad, Pakistan using inductively coupled plasma mass spectrometry.

The present study deals with the ground water quality assessment in Kahuta Industrial Triangle Islamabad, Pakistan. The objective of the study was to assess ground water quality against the drinking water standards for various toxic inorganic elements. Representative groundwater samples were collected and analyzed in the Water Quality Laboratory of Pakistan Council of Research in Water Resources (PCRWR) at Islamabad, Pakistan. The samples were run on ICP-MS (Inductively coupled plasma mass spectrometry), which has the capability to separate and quantify 70 elements at a time. One of the finding of study is that ICP-MS is a very good tool to analyze broad range of toxic inorganic elements to the level of parts per billion (ppb). World Health Organization drinking water standards shows that these toxic inorganic elements such as heavy metals even at this concentration level (ppb) are injurious to human health. This analysis indicated pollution of various toxic elements including Selenium. Vertical leachate through industrial waste septic tanks is identified as major cause of groundwater pollution in the Industrial Triangle. Monitoring of the septic tanks and groundwater quality in study area is suggested along with remedial measures.