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The DASH diet has been funded by the National Institute of Health (NIH) in various research projects to know whether specific dietary interventions helped treat HTN [ 30]. The subjects included in the study have been told to follow only dietary interventions and no other lifestyle modifications to avoid any confounding [ 30]. It was found in both hypertensive and normotensive individuals that dietary intervention alone helped reduce SBP by 6 to 11 mm Hg [ 30]. Since these results, the DASH diet, along with lifestyle modifications, has been advised as the first line of pharmacologic therapy in some instances. Several other clinical trials have shown that the DASH diet helps lower BP, cholesterol, and saturated fats as well as there is evidence that it lowers the risk of adverse cardiac events, type 2 diabetes, stroke, and obesity [ 30]. Therefore, it is crucial for clinicians, nurses, and pharmacists to educate patients about the benefits of the DASH diet [ 30]. 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. 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 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].
Centre for Ocean Science and Technology for Islands, National Institute of Ocean Technology, Ministry of Earth Sciences, Government of India, Port Blair, India
Conclusions
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 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. Enter [email protected] as the Account name and then enter or paste the manual entry code in from above as the 'Secret key'.
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 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 According to the National Institute on Alcohol Abuse and Alcoholism (NIAAA), low-risk drinking is considered four drinks per day or less than fourteen drinks per week for men and less than or equal to three drinks per day and less than seven drinks per day for women [ 49]. Binge drinking is the consumption of five drinks by men and four drinks by women in a period of two hours [ 47, 50]. According to studies performed by O’Keefe et al. and Peng et al., it was shown that excessive consumption of alcohol has contributed to 16% of HTN cases worldwide [ 45, 51]. The pathophysiology of HTN in chronic alcohol consumption is by vagal inhibition and sympathetic stimulation [ 52].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. 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.
Briasoulis 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]. 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 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].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
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]. 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). 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). Our preliminary study suggests that the detoxification of Hg 2+ is the immediate result of the GST and merA function as shown in Figure S6. The existence of the glutathione reductase gene in some bacterial mer operons also supports the role of LMW thiols in Hg 2+ detoxification ( Norambuena etal., 2018). The E. coli, together with the integrated merA - GST gene, was able to survive in the high Hg stress environment and transform Hg 2+ to Hg 0 ( Cursino etal., 2000). It is correlated that IOTG may act as Hg(II)-buffering agents and subsequently, Hg 2+ is reduced by merA. In the case of merA, mercuric ion was uptaken by the active process i.e. mer mediated transport (merP and merT) and converted to Hg 0 form by mercuric reductase. On the other hand, the Hg 2+ that was uptaken through the passive process could subsequently be inactivated by GST or conjugated to merA. Thus, an alternative method is proposed via which Hg resistance level may be augmented in bacteria: the sequestering of Hg given a protein-ligand interaction may lead to an improved way for the volatilization process. ConclusionsAssorted studies have shown that IF lowers BP. Harvie et al., in their study involving 107 overweight or obese premenopausal women, showed that IF for six months helped lower SBP (p= 0.99) and DBP (p= 0.84) [ 22]. Varady et al., in their study performed for 12 weeks with IF involving 15 overweight individuals (five males, 10 females with a BMI of 20-29.9 kg/m 2 showed that IF helped lower BP with a p-value of 0.51 [ 23]. A study by Bhutani et al. involving 83 obese individuals (three males and 80 females) with a BMI of 30-39.9 kg/m 2revealed that 12 weeks of IF helped lower SBP (p = 0.254) and DBP (p = 0.570) [ 24]. In a study by Eshghinia et al., 15 overweight or obese women with a BMI ≥25 kg/m 2 who followed IF for eight weeks showed a lowering of SBP (p<0.001) and DBP (p<0.05) [ 25]. Teng et al., in their 12 weeks IF study with 28 Malay men with a BMI of 23-29.9 kg/m 2,showed a lowering in SBP (p<0.05) and DBP (p<0.05) [ 26]. Erdem et al., in their study involving 60 participants from the Cappadocia cohort with pre-HTN and HTN with SBP of 120-139 and more than or equal to 140 mm Hg, DBP of 80-80 and more than or equal to 90 mm Hg, revealed that IF helps lower SBP (p<0.001) and DBP (p<0.039) [ 27]. 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. Although weight loss plays a significant role in lowering BP, it may be challenging to stay constantly motivated for long-term results [ 28]. Developing strategies to identify individuals who are unable to maintain lifestyle changes may be crucial to help them stay motivated to achieve weight management goals [ 28]. Furthermore, in recent times, an effective tool to promote the maintenance of healthy lifestyle changes like weight loss can be the use of mobile technology and personal digital devices, especially when individuals no longer have the availability of support and accountability through active interventions [ 29]. In hypertensive individuals with normal weight, other interventions like the DASH diet help lower BP [ 28]. 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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