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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 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 Weight loss is one of the most important non-pharmacological interventions to lower BP. Numerous interrelated pathophysiologic mechanisms stimulate higher BP in obesity [ 12]. In overweight/obese individuals, accelerated vascular aging can lead to HTN due to inflammation, oxidative stress, and insulin resistance [ 13]. Obese individuals also experience increased activity of the sympathetic nervous system and the renin-angiotensin-aldosterone system [ 14]. The combined effect results in increased sodium resorption by the kidney, impaired vasodilation, volume expansion, and decreased natriuresis, thus leading to elevated BP [ 15]. Ozemek et al. trials have shown that weight loss helped lower systolic BP in hypertensive individuals by 5 mm Hg and in normotensive individuals by 2 to 3 mm Hg. According to the Centers for Disease Control (CDC), a BMI of 25 to 29.9 is considered overweight, and a BMI of 30 or higher is considered obese. In overweight or obese individuals, achieving their ideal body weight is best, but it is good to aim for at least a 1 kg reduction in body weight. The study by Ozemek et al. also revealed that for every 1 kg reduction in body weight, we can expect about a 1 mm Hg reduction in blood pressure. 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). One of the quintessential steps to manage HTN is lifestyle modifications like exercise, weight loss, dietary interventions, a low-sodium diet, limiting alcohol consumption, smoking cessation, and stress management to help control BP. The treatment of HTN is based on specific characteristics like stage of disease, compliance, and comorbid conditions. For proper management of HTN, we may need to include pharmacological and non-pharmacological interventions [ 5]. Despite advancements in medicine and treatment options, the global burden of HTN has been increasing due to the advancing age of the population and an increasing prevalence of obesity. HTN is an insidious disease that, if not treated promptly, predisposes us to cardiovascular complications and various other complications [ 6]. Increased physical activity, limiting salt intake, minimizing alcohol consumption, smoking cessation, and stress management together support the management of patients with HTN and as preventive measures in the general population [ 7]. However, it is important to note that lifestyle modification is a process that requires patients to adhere continuously [ 5].
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 contributionsMercury (Hg) pollution poses a global threat to human and environmental health due to its noxiousness, mobility, and lengthy residence duration in the atmosphere ( Raphael etal., 2011). Toxic metals rapidly accumulate in the food chain, impacting higher trophic levels, hence these are of principal concern nowadays ( Raphael etal., 2011). According to recent research, oceanic release and biomass burning (organic compounds) account for the majority of worldwide Hg emissions, with anthropogenic activities accounting for the remaining significant percentage ( Pirrone etal., 2010; Nelson etal., 2012; Serrano etal., 2013). Hg exists in elemental, inorganic, and organic forms in both land and water systems, depending on oxidation-reduction conditions. In the atmosphere, Hg with valence +2 is more extensively spread ( Wang etal., 2004). In the biogeochemical cycle, a considerable part of Hg accumulates in seas and oceans by its atmospheric deposition ( Bindler, 2003; Wang etal., 2004). The effects of human-induced sources on the Hg contents and its forms are extensively larger in the photic layers of oceans ( Strode etal., 2007).
Based on the results obtained by the x-ray film method, ICP-MS analysis has been carried out to confirm the Hg removal potential of the isolates in the presence of 50 mg/L of Hg 2+. The ICP-MS outcomes showed significant changes in the level of Hg depletion. Among all, the marine culture NIOT-EQR_J251 possessed the highest potential (70.62%) to volatilize Hg. NIOT-EQR_J258 showed the least removal of Hg (17.48%), whereas NIOT-EQR_J7 and NIOT-EQR_J248 were capable to volatilize 29.18% and 52.17% of Hg 2+ under similar conditions, respectively. ( Figure4B). NIOT-EQR_J251 could volatilize 32.54 mg of the initial inoculum of 46.07 mg of HgCl 2, whereas NIOT-EQR_J258 removes 8.03 mg of the initial supplement of 45.74 mg of Hg 2+. In presence of 50 mg/L of Hg 2+, NIOT-EQR_J7 and NIOT-EQR_J248 dominantly developed the fog on the x-ray film, whereas, in the ICP-MS analysis, NIOT-EQR_J251 showed the highest Hg removal as compared to the other isolates. This suggests that volatilization was not solely responsible for removing the Hg 2+ from the culture media and the contribution of different processes like bioaccumulation and bio-adsorption may be involved. It can be observed that high Hg concentration causes structural deformities which may be due to its toxic nature. In our previous study, we reported that cultures accustomed to grow in the presence of HgCl 2 manifested major morphological abnormalities ( Joshi etal., 2021). We also suggested several structural asymmetries allied with the cell wall and cytoplasmic membrane, significantly affecting the cellular mechanism. Resultantly, there is a substantial delay at the beginning of growth and cell division in a higher concentration of Hg 2+. Metabolites formed during Hg (II) reduction Science and Technology for Islands, National Institute of Ocean Technology, Ministry of Earth Sciences, Government of India, Chennai, IndiaBriasoulis et al., in their meta-analysis of 16 prospective long-term studies, revealed that alcohol consumption of more than 20 g per day increases the risk of HTN significantly in women, whereas consumption of 31 to 40 g in men significantly increases the risk of HTN [ 54]. Nevertheless, a higher risk of HTN was observed in all individuals who consumed more than 20 g of alcohol per day, irrespective of their gender [ 54]. Ozemek et al. revealed in their study that, in persons who consume alcohol, a reduction to two standard drinks or less daily in men and one drink or less daily in women has been shown to reduce SBP by 4 mm Hg in hypertensive individuals and by 3 mm Hg in normotensive individuals [ 11]. 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]. According to a study by Ozemek et al., healthy diets like DASH dietary patterns have been shown to decrease SBP by 11 mm Hg in hypertensive individuals and by 3 mm Hg in normotensive individuals. Ideally, the goal is to reduce sodium to <1500 mg/dl, but it is good to aim for at least a 1000 mg/day reduction, which has been shown to reduce SBP approximately by 5/6 mm Hg in hypertensive individuals and by 2/3 mm Hg in normotensive individuals [ 11]. Furthermore, Ozemek et al. revealed in their study that increasing dietary potassium intake with a goal of 3500-5000 mg/day has been shown to reduce SBP by −4/5 mm Hg in hypertensive individuals and by −2 mm Hg in normotensive individuals [ 11]. 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 Enter [email protected] as the Account name and then enter or paste the manual entry code in from above as the 'Secret key'.
In prior studies, many pathways have been reported for Hg 2+ detoxification in microbes using various tools and technologies ( Chang etal., 2020; Cursino etal., 2000; Chang etal., 2021). In the present study, a metabolic pathway is hypothesized based on the GC-MS metabolites and protein-ligand interaction ( Figure S6) and an attempt has been made to correlate the proposed pathway with the different components of the already existing pathways. In this preliminary study, we found that general resistance/detoxification mechanisms of NIOT-EQR_J248 and NIOT-EQR_J251 in response to inorganic mercury (Hg 2+) exposure were a multisystem combined process. Based on phenotype characteristics, a total of 162 bacterial colonies grown on the ZMA media supplemented with 10 mg/L of Hg as HgCl 2 were selected for further analysis. In the presence of 25 mg/L of Hg 2+, only 63 isolates out of 162 showed resistance. Further, these 63 isolates were characterized based on their growth pattern in the presence of more than 25 mg/L i.e. 50, 75, and 100 mg/L of Hg 2+ concentration. Among the 63 isolates, 21 isolates grew in the presence of 50 mg/L, followed by 9 and 4 isolates at 75 and 100 mg/L of HgCl 2, respectively. The growth pattern of 4 isolates in the presence of HgCl 2 is shown in Figure S1. 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].
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 GJ: Conceptualization, Methodology, Formal analysis, Investigation, Writing-original draft, Writing-review & editing. PV: Investigation, Formal analysis, Writing-review & editing. BM: Investigation. PG: Formal analysis. DMP: Investigation. DKJ: Writing-review & editing. NVV: Supervision, Project administration, Writing-review & editing. GD: Supervision, Project administration, Writing-review & editing. All authors contributed to the article and approved the submitted version. Funding Three experimental replicates were determined for each analysis and the statistical significance of the results was analyzed by one way ANOVA test. All Statistical analyses were carried out using SPSS (version 17) and GraphPad Prism 7 software. Differences were considered significant at p< 0.05 and values are reported as mean ± standard deviation (SD). Results and discussion
The GC-MS study was included to get insight into the metabolic changes in the MRB isolates under Hg stress. It revealed that the metabolic profiles of MRB isolates are diverse with different biological properties. GC-MS metabolic profiles revealed that the number of compounds increased in the presence of Hg compared to the control isolates, where some compounds were found to be different in Hg-treated samples. A total of 50 metabolites were identified in the absence of Hg, whereas with 50 mg/L of Hg, 64 compounds were identified. The minimum metabolites (6) were identified in NIOT-EQR_J258 without Hg, whereas the maximum metabolites (22) was identified in NIOT-EQR_J251 with Hg 2+. The NIOT-EQR_J7 metabolic profile was almost the same in both control and Hg-treated samples. A total of 14 metabolites were present in the control sample, whereas 15 metabolites were identified with Hg. In the case of NIOT-EQR_J248, NIOT-EQR_J251, and NIOT-EQR_J258, a total of 10, 20, and 6 compounds were identified in the control sample, whereas 16, 22, and 11 metabolites were present with Hg, correspondingly. 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]. 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.
References
The following four randomized controlled trials have been conducted to show that exercise helped lower both systolic and diastolic BP in participants: In the meta-analyses of randomized controlled trials conducted by Fagard et al., 72 trials have been conducted with an average of 40 participants per trial, which involved a 16-week study of 40 minutes of exercise sessions three times/week with an average intensity of 65% of heart rate [ 39]. This study showed a decrease in SBP of 6.9 mm Hg and a lowering of DBP of 4.9 mm Hg [ 39]. In 27 randomized controlled trials by Lee et al. with 1842 participants, the exercise regimen involved walking for 26.5 min/day for 4.4 days/week for a mean of 19 weeks [ 40]. This study shows that there is a larger effect with more intense and frequent exercise regimens for a longer duration [ 40]. There was a mean decrease in SBP of 5.2 to 11 mm Hg and in DBP of 3.8 to 7.7 mm Hg [ 40]. In the meta-analysis of randomized control trials performed by Cornelissen et al., 15 trials with 633 participants involving the exercise of 30-60 min, two to five times/week, at 50% to 75% HR reserve for six to 52 weeks showed a daytime decrease in SBP of 3.2 mm Hg and in DBP of 2.7 mm Hg [ 41]. However, no blood pressure reduction was seen at night [ 42]. The meta-analysis of randomized controlled trials performed by Cornelissen and Smart included 105 trials with 3957 participants [ 42]. This study concluded that moderate aerobic exercise involving walking and jogging for 30 to 60 min/session three to five times/week for four to 52 weeks showed a reduction in SBP of 3.5 mm Hg and in DBP of 2.5 mm Hg [ 42]. 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].
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