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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 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 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 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 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 identification

Science and Technology for Islands, National Institute of Ocean Technology, Ministry of Earth Sciences, Government of India, Chennai, India

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 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 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. 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.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 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 selected isolates were further analyzed by subjecting them to 16S rDNA sequencing. In brief, DNA was isolated and the polymerase chain reaction (PCR) amplification was executed with the universal 16S rDNA primers of 27F (5’-AGAGTTTGATCMTGGCTCAG-3’) and 1495R (5’-GGHTACCTTGTTACGACTT-3’) as described by Dash and Das (2014). The amplified PCR products were sequenced and submitted to NCBI, as described by Kumar etal. (2018). The detailed procedure is elaborated in the SI. Genotyping of merA gene