CRISPR-Cas9: Role in Processing of Modular Metabolic Engineered Bio-Based Products

Biogenetic engineering is a significant technology to sensibly manage microbial metabolic product factories. Genome modification methods for efficiently controlling and modifying genes at the genome level have progressed in biogenetic engineering during the last decade. CRISPR is genome editing technology that allows for the modification of organisms’ genomes. CRISPR and its related RNA-guided endonuclease are versatile advanced immune system frameworks for defending against foreign DNA and RNAs. CRISPR is efficient, accessible, and trustworthy genomic modification tool in unparalleled resolution. At present, CRISPR-Cas9 method is expanded to industrially manipulate cells. Metabolically modified organisms are quickly becoming interested in the production of different bio-based components. Here, chapter explore about the control productivity of targeted biomolecules in divergent cells based on the use of different CRISPR-related Cas9

Drug Utilization Study in Patients Attending Hypertension Clinic of a Tertiary Care Hospital, Rajasthan, India

Background: Hypertension is a major public health problem in India, affecting a significant proportion of the population. Drug utilization research is important for assessing the rationality of drug treatment and for identifying areas for improvement. This study examined the drug utilization pattern of antihypertensive drugs in hypertensive patients at an Outpatient Department (OPD) in Udaipur, Rajasthan, India, based on JNC-8 classification. Materials and Methods: The observational cross-sectional study included 300 hypertension patients aged above 25 attending the hypertension clinic. Exclusions were made for inpatients, age < 25, uncertain diagnosis, pregnant/lactating mothers, and patients subsequently admitted after OPD visits. Results: The majority of affected patients were above 60 years old, with most having hypertension for 2-5 years, often accompanied by Type II Diabetes Mellitus. Oral administration was the primary drug delivery route. On average, patients received 1.93 antihypertensive drugs per encounter (range: 1 to 4), with an average of 5.82 drugs per encounter (range: 1 to 12). Losartan (72%) and amlodipine (46%) were the most prescribed drugs, with Angiotensin Receptor Blockers (ARBs) being the most prescribed drug class (79.3%), followed by Calcium Channel Blockers (CCBs) at 46.6%. The most common therapy was a two-drug combination (44.6%), followed by single-drug therapy (33%). Approximately 26.3% of prescriptions indicated potential drug interactions. Conclusions: This study provides valuable baseline data on the prescribing pattern of antihypertensive drugs. Rational prescribing practices are being followed, with the majority of patients being prescribed a combination of two or three antihypertensive drugs. However, there is a need to educate patients about the risks associated with uncontrolled high blood pressure and the benefits of lifestyle changes

Synergistic Activity of Rhamnolipid Biosurfactant and Nanoparticles Synthesized Using Fungal Origin Chitosan Against Phytopathogens

Phytopathogens pose severe implications in the quantity and quality of food production by instigating several diseases. Biocontrol strategies comprising the application of biomaterials have offered endless opportunities for sustainable agriculture. We explored multifarious potentials of rhamnolipid-BS (RH-BS: commercial), fungal chitosan (FCH), and FCH-derived nanoparticles (FCHNPs). The high-quality FCH was extracted from Cunninghamella echinulata NCIM 691 followed by the synthesis of FCHNPs. Both, FCH and FCHNPs were characterized by UV-visible spectroscopy, DLS, zeta potential, FTIR, SEM, and Nanoparticle Tracking Analysis (NTA). The commercial chitosan (CH) and synthesized chitosan nanoparticles (CHNPs) were used along with test compounds (FCH and FCHNPs). SEM analysis revealed the spherical shape of the nanomaterials (CHNPs and FCHNPs). NTA provided high-resolution visual validation of particle size distribution for CHNPs (256.33 ± 18.80 nm) and FCHNPs (144.33 ± 10.20 nm). The antibacterial and antifungal assays conducted for RH-BS, FCH, and FCHNPs were supportive to propose their efficacies against phytopathogens. The lower MIC of RH-BS (256 μg/ml) was observed than that of FCH and FCHNPs (>1,024 μg/ml) against Xanthomonas campestris NCIM 5028, whereas a combination study of RH-BS with FCHNPs showed a reduction in MIC up to 128 and 4 μg/ml, respectively, indicating their synergistic activity. The other combination of RH-BS with FCH resulted in an additive effect reducing MIC up to 128 and 256 μg/ml, respectively. Microdilution plate assay conducted for three test compounds demonstrated inhibition of fungi, FI: Fusarium moniliforme ITCC 191, FII: Fusarium moniliforme ITCC 4432, and FIII: Fusarium graminearum ITCC 5334 (at 0.015% and 0.020% concentration). Furthermore, potency of test compounds performed through the in vitro model (poisoned food technique) displayed dose-dependent (0.005%, 0.010%, 0.015%, and 0.020% w/v) antifungal activity. Moreover, RH-BS and FCHNPs inhibited spore germination (61–90%) of the same fungi. Our efforts toward utilizing the combination of RH-BS with FCHNPs are significant to develop eco-friendly, low cytotoxic formulations in future

A prospective study evaluating the effect of a ‘Diagnostic Stewardship Care-Bundle’ for automated blood culture diagnostics

ABSTRACT: Objectives: We prospectively implemented a diagnostic stewardship care-bundle checklist, ‘Sepsis-48 DSB’, with the aim of reducing intervening duration of key steps of automated blood culture diagnostics (aBCD). Methods: Sepsis-48 DSB was implemented for automated blood culture bottles (BCBs) received from adult intensive care units (AICUs) during the intervention period (P2; July 2020–June 2021) and intervening durations were compared with those during the retrospective, pre-intervention period (P1; March–June 2020). During both periods, provisional blood culture reports (pBCR) were issued wherein direct microbial identification (dID) was performed in BCBs with Gram-negatives by directly inoculating conventional biochemical tests and direct antimicrobial susceptibility testing (dAST) using EUCAST RAST method. The results were compared with the standard of care (SoC) method (i.e. full incubation followed by identification and AST by VITEKⓇ-2 Compact). Results: During P2, significant reductions in loading time (LT) [median: 63.5 vs. 32 minutes, P < 0.001], time to dID+dAST performance (TTD) [186 vs. 115 minutes, P = 0.0018] and an increase in compliance to bundle targets [LT ≤45: 44% vs. 66%, P = 0.006 and TTD ≤120: 34% vs. 51.7%, P = 0.03] were observed. Using dID+dAST method, results were read 694 minutes earlier than SoC method. Of 176 pBCR, 165 (94%) were concordant with SoC in microbial identification of species. Categorical agreement for any drug-bug combination was 92.7% (1079/1164) and corresponding major, very major, and minor error rates were 8.8% (19/216), 4.9% (45/921), and 1.8% (21/1164), respectively. Conclusion: The ‘diagnostic stewardship care-bundle’ strategy was successfully implemented with considerable diagnostic accuracy leading to significant reductions in duration of targeted steps of aBCD

A prospective study evaluating the effect of a “Diagnostic Stewardship Care-Bundle” for automated blood culture diagnostics

ABSTRACT: Objectives: We prospectively implemented a diagnostic stewardship care-bundle checklist, ‘Sepsis-48 DSB’, with the aim of reducing intervening duration of key steps of automated blood culture diagnostics (aBCD). Methods: Sepsis-48 DSB was implemented for automated blood culture bottles (BCBs) received from adult intensive care units (AICUs) during the intervention period (P2; July 2020–June 2021) and intervening durations were compared with those during the retrospective, pre-intervention period (P1; March–June 2020). During both periods, provisional blood culture reports (pBCR) were issued wherein direct microbial identification (dID) was performed in BCBs with Gram-negatives by directly inoculating conventional biochemical tests and direct antimicrobial susceptibility testing (dAST) using EUCAST RAST method. The results were compared with the standard of care (SoC) method (i.e. full incubation followed by identification and AST by VITEKⓇ-2 Compact). Results: During P2, significant reductions in loading time (LT; median: 63.5 vs. 32 minutes, P < 0.001), time to dID+dAST performance (TTD; 186 vs. 115 minutes, P = 0.0018) and an increase in compliance to bundle targets (LT ≤45: 44% vs. 66%, P = 0.006 and TTD ≤120: 34% vs. 51.7%, P = 0.03) were observed. Using dID+dAST method, results were read 694 minutes earlier than SoC method. Of 176 pBCR, 165 (94%) were concordant with SoC in microbial identification of species. Categorical agreement for any drug-bug combination was 92.7% (1079/1164) and corresponding major, very major, and minor error rates were 8.8% (19/216), 4.9% (45/921), and 1.8% (21/1164), respectively. Conclusion: The ‘diagnostic stewardship care-bundle’ strategy was successfully implemented with considerable diagnostic accuracy leading to significant reductions in duration of targeted steps of aBCD