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Increased physical activity has been advocated as the first-line intervention for preventing and treating patients with prehypertension and as a treatment strategy for patients with stage 1 or stage 2 HTN, according to the Duthe American College of Sports Medicine, the United States Joint Nations Committee on Prevention, Detection, Evaluation, and Treatment of High BP, the World Health Organization and International Society of Hypertension, and The National Heart Foundation [ 36]. Exercise can consequently help prevent prehypertension from progressing and can help reduce or stop medications prescribed for the treatment of stage 1 HTN [ 37]. A conserved region of merA gene encoding bacterial mercuric reductase enzyme was detected using the above isolated genomic DNA ( Joshi etal., 2021). The PCR reactions were carried out using the primer set F1merA (5’-TCGTGATGTTCGACCGCT-3’) and F2merA (5’-TACTCCCGCCGTTTCCAAT-3’) containing 1 U/μl Taq polymerase (Sigma-Aldrich), 1 × Enzyme buffer, 200 μM of each dNTP (Sigma-Aldrich), 1.25 mM MgCl 2, and 0.5 μM of each primer and 50 ng template DNA in a thermal cycler (Applied Biosystems). PCR reaction was carried out under the following amplification conditions: pre-denaturation step at 94°C for 2 min followed by 30 cycles of 94°C for 1 min, 52°C for 1 min and an extension step at 72°C for 1 min and final extension at 72°C for 7 min as described by Dash and Das, (2014). For the isolates that showed negative PCR reactions with F1merA and F2merA primer set, a re-attempt has been made for amplifying the merA gene by gradient PCR method (annealing at 54 ± 5°C) and by using another set of primer A1F (5′-ACCATCGGCGGCACCTGCGT-3’) and A5R (5′-ACCATCGTCAGGTAGGGGACCAA3-′) as described by De etal. (2008). Amplification of the merA gene was confirmed by visualizing it under the Gel Doc system (BioRad). Confirmation of Hg 2+ to Hg 0 reduction by the isolates The modification in functional groups present in the culture pellets was identified by measuring the spectra in the range of 400 to 4000 cm -1 using Fourier transform infrared (FT-IR) spectroscopy (IR Affinity-I spectrometer, Shimadzu, Japan), as described by Joshi etal. (2021). In brief, 48 h grown cultures in ZMB medium without Hg supplementation were used as a control, whereas cultures with 50 mg/L of Hg supplementation were used as Hg treated. The mixtures (lyophilized cells and 2% KBr) were fixed in the FT-IR spectrometer after compressing them into translucent sample discs, followed by analyzing in ATR-FT-IR mode by following the manufacturer’s protocol. Scanning Electron Microscopy (SEM) analysis of MRB In context to these four isolates, the shift in wavenumbers, changes in peak height, and the appearance/disappearance of new peaks in the presence of Hg 2+; suggest alterations in functional groups (especially -SH group), which might play an important role in the Hg detoxification. SEM analysis of Hg treated and untreated isolates
Hg volatilization by bacteria involves the reduction of Hg 2+ via the mer system to Hg 0, which is less toxic. It is appealing from the essential and practical viewpoints that halophilic bacteria can also grow in the presence of a higher amount of Hg and volatilized Hg efficiently similar to the non-halophilic bacteria. The ICP-MS analysis did not detect significant Hg loss in the control sample (without bacteria), which confirms the volatility of Hg being biotic ( Table S2). Thus, our results suggested that halophilic bacteria can also remove the Hg from the growth media, indicating that it may be potentially applied in contaminated environments. The Hg comes into the ocean either through atmospheric deposition or other several natural and active or passive processes. The results of this study imply that the two marine strains of Halomonas sp., isolated from ERIO may have the ability to remove Hg 2+ from the growth medium. Identification of functional groups changed during Hg(II) reduction The statistical analysis of Hg removal potential among the different species i.e., Alcanivorax xenomutans, Halomonas sp., Marinobacter hydrocarbonoclasticus and within the Halomonas spp. revealed that the Hg removal by NIOT-EQR_J251 was significantly higher (p< 0.01) than NIOT-EQR_J7 and NIOT-EQR_J258. There was a significant difference (p< 0.05) in the removal of Hg between NIOT-EQR_J248 vs. NIOT-EQR_J258 and NIOT-EQR_J248 vs. NIOT-EQR_J251. Al-Mailem etal. (2011) reported Haloferax sp. (HA1 and HA2), Halobacterium sp. HA3, and Halococcus sp. HA4 effectively volatilized (from 40 to 65%) the available 100 mg/L of Hg after 8 days. Many other isolates such as Bacillus sp., Pseudomonas stutzeri, Pseudomonas putida, Vibrio fluvialis could volatilize 60%-95%, 94%, 100%, 60% of Hg 2+, respectively, from culture ( Zhang etal., 2012; Dash etal., 2013; Giri etal., 2014; Saranya etal., 2017; Zheng etal., 2018).Now the top of the interior is clean, it's time to shake and beat the mats. That will deal with the larger bits of dirt. HG car dashboard cleaner makes the interior of the car look like new. This dash cleaner cleans and protects synthetic materials, plastic, rubber, vinyl and leather. Also, the car dash cleaner intensifies the colour and restores the shine of the polished surface. The HG dashboard spray also leaves a wonderfully fresh fragrance after the treatment, but not a detectable greasy coating. How do you use HG car dashboard cleaner?
The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/ Supplementary Material. Author contributions Genus Halomonas is reported from most of the saline environments regardless of their geographical location, including the marine environment, salterns, saline lakes, and soil ( Llamas etal., 2006). The MRB isolated from the coastal areas of Kuwait were identified as Alcanivorax borkumensis, Marinobacter hydrocarbonoclasticus, and Halomonas taeheungii ( Sorkhoh etal., 2010). The 16S rDNA analysis and phenotypic characteristics revealed that the heavy metal-resistant halophilic bacteria WQL9 belong to the genus Halomonas sp ( Abdel-Razik etal., 2020). The Marinobacter genus is comprised of widespread marine bacteria found exceptionally in marine or terrestrial environments rich in sodium salts, in the deep sea, coastal seawater, sediment, oceanic basalt, etc. ( Handley and Lloyd, 2013). Alcanivorax xenomutans are also rich in the saline environment as they are halophilic and favorable to living in the marine environment. A comparative analysis of the partial sequence of the 16S rDNA of the four strains revealed a high level of similarity to the corresponding sequence of environmental isolates. Detection of mercury reductase gene GC-MS analysis was performed to characterize the bacterial response and the metabolomic changes leading to Hg tolerance. The solvent extraction method was used to extract the bacterial metabolites. In brief, the freshly inoculated and exponentially grown MRB and MMRB bacterial cells in ZMB medium (non-exposed and exposed to 50 mg/L of Hg) were freeze-dried using BENCHTOP lyophilizer (VIRTIS Instrument, Gardiner, NY). For extraction of compounds, 50 mg of lyophilized bacterial cells were suspended in ethyl acetate and chloroform (1:1; v/v) and homogenized. After homogenization, the solution (crude extract) containing the metabolites was transferred to the clean glass vial by pipetting. These steps were repeated two-three times to obtain a pure and ample amount of sample. The separated organic fractions (crude extract) were treated with anhydrous NaSO 4 (Sigma-Aldrich) to remove moisture, which was again concentrated on the rotary evaporator (BUCHI Rotavapor R-215/V advanced, Switzerland) at RT and stored at -80°C until further analysis. The concentrated crude extract was re-suspended in 1 mL of Dichloromethane (DCM) and 5 µL of the sample was injected into the GC-MS analyzer (Agilent Technologies Instrument 7890A GC System, 240 Ion Trap GC/MS, USA). The GC-MS analysis was carried out under external ionization mode using a fused silica column HP 5 MS column (30 m × 0.320 mm × 0.25 µm). High purity helium was used as a carrier gas at a constant flow rate of 1 mL/min. For analysis, the chromatographic conditions i.e. initial injector and detector temperature, were set at 250°C and 330°C, respectively. The temperature of the column was programmed from 50°C (hold for 2 min) to 320°C (2 min hold), with a constant 5°C increment per minute and 1 min hold at 330°C. A metabolic library of all the separated compounds found via GC-MS analysis of bacterial extract was created and identified using NIST mass spectral library match. The PubChem CID, structures, names, and molecular weight of those bioactive compounds were obtained from the PubChem database. In-silico analysis Spray the HG car dashboard cleaner thinly and evenly onto the surface to be treated. Keep the can upright and approximately 20cm away from the dashboard.
How do you use HG car dashboard cleaner?
Patients can find it increasingly difficult to maintain long-term dietary changes like low caloric intake, low salt intake, and limiting processed foods, which in turn may lead to a shift towards preintervention practices in some individuals, for example, increased calorie intake [ 28]. This implies the need for interventions to focus on ways to help patients maintain healthy dietary patterns [ 28]. The Mediterranean diet (Med Diet) has also helped lower BP. As per the Maine-Syracuse Longitudinal Study conducted in the United States in 2020 by a group of researchers who followed 851 US older adults, for every one unit increase in the Med Diet score in participants, it was found that there was a corresponding reduction of 0.69 units in SBP, a reduction of 0.33 in DBP, and a reduction of 0.45 on mean arterial pressure (MAP) [ 32]. Although this seems to be small, this change can have a noteworthy effect at the level of the population; that is, a decrease of 2 mm Hg in SBP can lead to a decrease of 10% when it comes to the population [ 32]. According to the observational studies conducted in Mediterranean countries, higher adherence to a Med Diet is associated with a decreased risk of cardiovascular disease, overall mortality as well as neoplastic disease [ 33, 34]. The Med Diet consists of higher consumption of extra virgin olive oil, vegetables, fruits, whole grains, nuts, cereals, as well as seeds; moderate consumption of fish, poultry, red wine, and dairy; and lower consumption of processed foods and red meat [ 35]. Science and Technology for Islands, National Institute of Ocean Technology, Ministry of Earth Sciences, Government of India, Chennai, India
Hg tolerance of CMRBs was calculated by the broth dilution method ( Konopka and Zakharova, 1999; Santos-Gandelman etal., 2014) with some modifications. CMRB cultures grown in Zobell Marine Broth medium (ZMB, Himedia, Mumbai) without Hg supplementation were used as a positive control, whereas ZMB treated with Hg (without bacteria) was used as a negative control in this study. To ensure the uniform bioavailability of Hg to the bacteria, uniform culture conditions were maintained across different experimental treatments. Based on the growth behavior of cultures in the presence of a higher concentration of HgCl 2, the selected isolates were classified into two different categories i.e. MRB and moderate MRB (MMRB). The detailed procedure is elaborated in the SI. To understand the effects of different Hg concentrations on bacterial growth, two qualitative analyses were carried out, i.e. monitoring the optical density (OD) at 600 nm and dry biomass. 16S rDNA based identificationTo inspect the consequence of Hg 2+ concentration on the morphology, the bacterial cells were freshly grown in the presence (50 and 100 mg/L - test samples) and absence (control) of Hg 2+ as HgCl 2; and cell pellets were harvested by centrifugation (5000 rpm at 4°C for 5 min) after 48 h. SEM analysis has been carried out, as reported by Joshi etal. (2021). In brief, the bacterial cells were fixed with 2.5% glutaraldehyde followed by the post-fixing in 1% osmium tetroxide and dehydration of the cells with graded ethanol series (25%, 50%, 80%, 90%, and absolute). The processed samples were scanned using SEM (JEOL-JSM-IT500). Extraction of metabolites and Gas Chromatography-Mass Spectrometry (GC-MS) analysis The pathogenesis of HTN involves oxidative stress. Another mechanism involved is the decreased bioavailability of nitric oxide (NO) [ 38]. Physical exercise could be a potential lifestyle intervention to treat HTN due to its beneficial effects on endothelial function and oxidative stress [ 38]. Exercise exerts an anti-inflammatory action via the hypothalamic-pituitary-adrenal axis and via the sympathetic nervous system, thus affecting BP directly [ 9]. The physiologic effects of exercise are further divided into acute, post-exercise, and chronic [ 9]. Aerobic exercises like speed walking, jogging, running, cycling, dancing, and swimming have been shown to decrease resting BP and BP reactivity to stressors [ 9]. A study by Ozemek et al. revealed the following about how diverse types of exercises affect BP: (1) Aerobic exercise of 90 to 150 minutes per week with 65%-75% heart rate reserve has been shown to impact SBP by −5/8 mm Hg in hypertensive individuals and by −2/4 mm Hg in normotensive individuals. (2) Dynamic resistance exercise of 90 to 150 minutes per week with 50%-80% one rep maximum, six exercises, three sets/exercise, and ten repetitions/set has been shown to decrease SBP by 4 mm Hg in hypertensive individuals and 2 mm Hg in normotensive individuals. (3) Isometric resistance exercise of 4 × 2 min (hand grip), 1 min rest between exercises, 30%-40% maximum voluntary contraction, and three sessions per week for 8-10 weeks have been shown to lower SBP by 5 mm Hg in hypertensive individuals and 4 mm Hg in normotensive individuals [ 11]. It will have your employee’s name as held in the Payroll system, associated with this 'HG' account: Manual entry code method A simplified X-ray film method was adopted to look into the Hg reduction by bacterial volatilization in the presence of 50 and 100 mg/L of Hg as HgCl 2 ( Nakamura and Nakahara, 1988; Joshi etal., 2021). Briefly, the bacterial cells were collected by centrifugation at 5,000 rpm for 10 min. and washed with 0.07 M phosphate buffer (0.5 mM EDTA, 0.2 mM magnesium acetate, 5 mM sodium thioglycolate; pH 7.0) and transferred to the microplate. The cells were suspended into 50 µL of 0.07 M phosphate buffer containing 50 and 100 mg/L of Hg as HgCl 2 in a microplate. The phosphate buffers (pH 7.0) with 50 and 100 mg/L of HgCl 2 (without bacterial cells) were used as a negative control. The plate was covered with X-ray film and incubated in the dark for 2 h at 35°C. Determination of Hg(II) depletion potential by isolates The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Publisher’s note
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