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According to a study performed by Fuchs et al. in 2021, it has been found that, especially in women, the risk has been found to start at moderate alcohol consumption, and alcohol withdrawal has been found to promote the reduction in BP in short-term trials [ 52].Suliga et al., in their cross-sectional study of 12,285 individuals in the age group 37 to 66, described that daily alcohol consumption ranging from 0.1 g to 15 g was inversely related to the development of HTN in women, with an odds ratio of 0.67, 95% confidence interval of 0.59 to 0.75, p-value <0.001 [ 53]. However, in men, this relation was not observed [ 53]. Many studies suggest that mer operon is located on any one of either location such as mobile elements like plasmids, transposons, genomic or chromosomal DNA ( Liebert etal., 1999; Schelert etal., 2004; Zeng etal., 2010). The mer operon contains many functional genes along with the operator, promoter, and regulatory elements. All functional genes encrypt for the diverse proteins that contribute to the detection, transportation, and reduction of Hg ions ( Barkay etal., 2003; Dash and Das, 2012; Naguib etal., 2018). An extremely noteworthy positive correlation was found among the harboring of mer gene with the phenotypic resistance to Hg and concentration of the Hg in the source environment ( Osborn etal., 1997). In this study, we did not find any Hg from the ecosystem (detectable limit - 0.73 µg/L), but the amplification of mer genes and available literature suggested that these are extensively dispersed in a non-highly contaminated or contamination-free environment such as an open ocean, Antarctic sea-ice, high Arctic snow, sea ice brine etc. and plays a key role in the biogeochemical cycle of Hg ( Christakis etal., 2021). Thus, the Hg adaptation by these isolates is reasonably accepted. Confirmation of Hg 2+ reduction by the isolates 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).
Science and Technology for Islands, National Institute of Ocean Technology, Ministry of Earth Sciences, Government of India, Chennai, India To 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 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 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]. According to the American College of Cardiology, a BP higher than 180/120 mm Hg is considered a hypertensive emergency or crisis. Patients with these blood pressures need emergency medical help. Untreated high BP may increase the risk of myocardial infarction, stroke, and other serious complications. Monitoring BP every two years, starting at age 18, is important to diagnose and treat hypertension timely to prevent complications. HTN is diagnosed by performing repeated careful measurements of blood pressure. Blood pressure is categorized as follows: Normal blood pressure, defined as systolic blood pressure (SBP) less than 120, and diastolic blood pressure (DBP) less than 80. An elevated BP is an SBP of 120 to 129 and a DBP of less than 80. HTN is defined as a systolic pressure more than or equal to 130 or a diastolic pressure more than or equal to 80.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 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 Physicians play a key role in encouraging and helping patients achieve smoking cessation [ 56]. HTN has also been associated with second-hand smoke. According to research done by Bernabe-Ortiz et al. on 897 individuals in Peru in 2021 to assess the association of second-hand smoke with HTN and cardiovascular risk, 15% of adults reported second-hand smoke overall, and this emphasizes the necessity to keep places smoke-free to reduce the risk of cardiovascular disease [ 57]. 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.
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 upholstery suffers most. We have HG car upholstery cleaner. Or take a look at our tips for cleaning fabric car upholstery. For leather seats, we have HG deep cleaner for leather. The health effects of alcohol intake are variable and are based on the amount of intake (low, moderate, or heavy) and intake pattern (acute, chronic, or binge) [ 45- 47]. In the US, the amount of pure alcohol in one standard drink is 14 g, which can be found in 12 oz of regular beer (about 5% alcohol), 5 oz of wine (about 12% alcohol), and 1.5 oz of distilled spirits (about 40% alcohol) [ 48]. Moderate alcohol intake recommendations according to the Dietary Guidelines for Americans are the consumption of two standard drinks per day for men and one standard drink per day for women [ 49]. Moderate alcoholic intake has been shown to reduce the risk of chronic disease [ 49].Intermittent fasting (IF) is an effective way to lose weight and thus helps lower blood pressure [ 10]. The mechanism by which IF lowers BP may be due to a brain-derived neurotrophic factor (BDNF)-induced increase in parasympathetic activity [ 16]. Increased excretion of norepinephrine and increased sensitivity of insulin and natriuretic peptides also play a role [ 16]. The activation of glutamatergic receptors produces BDNF [ 10]. IF also stimulates the release of BDNF [ 10]. BDNF, in turn, stimulates the cholinergic neurons to release acetylcholine, which via the vagus nerve, controls the cardiac function to the sinoatrial (SA) node, causing a reduction of heart rate [ 17]. Also, blood vessels are expanded by the neurotransmitter, leading to a reduction in BP [ 18]. The pathogenesis of blood pressure lowering by activation of the parasympathetic nervous system involves the role of the cerebrospinal stem in the activation of cholinergic neurons [ 19, 20]. However, cardiovascular health benefits have only been observed to last as long as the IF diet lasted and pressures returned to initial values after the completion of the IF diet [ 21]. Toledo et al. performed a study in Germany in which 1422 participants on the IF diet were followed up for one year [ 16]. These participants had a fasting period of four to 21 days, which involved 200-250 kcal daily meals [ 16]. In participants who fasted for a longer time, a reduction of SBP and DBP was observed [ 16].
In our recent study ( Joshi etal., 2021), we isolated and characterized several MRBs from the deeper depth of the Central Indian Ocean and evaluated their ability to remove Hg from the culture media. However, no bacterial strain was observed from the surface seawater samples that were resistant to Hg up to 100 mg/L. Though various bacterial strains have been isolated from the coastal and oceanic region, so far. MRB from the equatorial region of the Indian Ocean (ERIO) is not studied. It is hypothesized that the marine bacteria from ERIO could be a potential resource for the reduction of Hg 2+ as ERIO is highly dynamic in nature due to high current and intense climatological precipitation ( Annamalai, 2010).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
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