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Technologies capable of monitoring product quality attributes and process parameters in real time are becoming popular due to the endorsement of regulatory agencies and also to support the agile development of biotherapeutic pipelines. The utility of vibrational spectroscopic techniques such as Fourier transform mid-infrared (Mid-IR) and multivariate data analysis (MVDA) models allows the prediction of multiple critical attributes simultaneously in real time. This study reports the use of Mid-IR and MVDA model sensors for monitoring of multiple attributes (excipients and protein concentrations) in real time (measurement frequency of every 40 s) at ultrafiltration and diafiltration (UF/DF) unit operation of biologics manufacturing. The platform features integration of fiber optic Mid-IR probe sensors to UF/DF set up at the bulk solution and through a flow cell at the retentate line followed by automated Mid-IR data piping into a process monitoring software platform with pre-loaded partial least square regression (PLS) chemometric models. Data visualization infrastructure is also built-in to the platform so that upon automated PLS prediction of excipients and protein concentrations, the results were projected in a graphical or numerical format in real time. The Mid-IR predicted concentrations of excipients and protein show excellent correlation with the offline measurements by traditional analytical methods. Absolute percent difference values between Mid-IR predicted results and offline reference assay results were ≤5% across all the excipients and the protein of interest; which shows a great promise as a reliable process analytical technology tool.

Artificial intelligence (AI) is a huge headway in innovation that has everybody talking about its energizing guarantees in the innovation world. With regards to AI, it additionally incorporates its territories, for example, AI (ML). While AI could be portrayed as the capacity of machines to settle on shrewd human-like choices and improve over the long run, ML includes building models, generally statistical models that give prescient outcomes and can be developed. Many are not extremely educated about this area. While this is true, there is something else entirely to it from face recognition, fingerprints recognition, chat-bots, predictive business models, and sentimental analysis. Beforehand, AI joining in the product advancement was simply conceivable to the huge organizations that had the assets to recruit exceptionally qualified experts. Over the long run, AI structures with high deliberation levels have been created, and with few coding lines in any programming language of the decision, one can have the option to enter in various fields.

In many bacteria and eukaryotes, replication fork establishment requires the controlled loading of hexameric, ring-shaped helicases around DNA by AAA+(ATPases Associated with various cellular Activities) ATPases. How loading factors use ATP to control helicase deposition is poorly understood. Here, we dissect how specific ATPase elements ofEscherichia coliDnaC, an archetypal loader for the bacterial DnaB helicase, play distinct roles in helicase loading and the activation of DNA unwinding. We have identified a new element, the arginine-coupler, which regulates the switch-like behavior of DnaC to prevent futile ATPase cycling and maintains loader responsiveness to replication restart systems. Our data help explain how the ATPase cycle of a AAA+-family helicase loader is channeled into productive action on its target; comparative studies indicate that elements analogous to the Arg-coupler are present in related, switch-like AAA+ proteins that control replicative helicase loading in eukaryotes, as well as in polymerase clamp loading and certain classes of DNA transposases.

The biopharmaceutical industry is transitioning from currently deployed batch-mode bioprocessing to a highly efficient and agile next-generation bioprocessing with the adaptation of continuous bioprocessing, which reduces capital investment and operational costs. Continuous bioprocessing, aligned with FDA's quality-by-design platform, is designed to develop robust processes to deliver safe and effective drugs. With the deployment of knowledge-based operations, product quality can be built into the process to achieve desired critical quality attributes (CQAs) with reduced variability. To facilitate next-generation continuous bioprocessing, it is essential to embrace a fundamental shift-in-paradigm from "quality-by-testing" to "quality-by-design," which requires the deployment of process analytical technologies (PAT). With the adaptation of PAT, a systematic approach of process and product understanding and timely process control are feasible. Deployment of PAT tools for real-time monitoring of CQAs and feedback control is critical for continuous bioprocessing. Given the current deficiency in PAT tools to support continuous bioprocessing, we have integrated Infinity 2D-LC with a post-flow-splitter in conjunction with the SegFlow autosampler to the bioreactors. With this integrated system, we have established a platform for online measurements of titer and CQAs of monoclonal antibodies as well as amino acid analysis of bioreactor cell culture.

Process analytical technology (PAT) is a fast-growing field within bioprocessing that enables innovation in biological drug manufacturing. This study demonstrates novel PAT methods for monitoring multiple quality attributes simultaneously during the ultrafiltration and diafiltration (UF/DF) process operation, the final step of monoclonal antibody (mAb) purification. Size exclusion chromatography (SEC) methods were developed to measure excipients arginine, histidine, and high molecular weight (HMW) species using a liquid chromatography (LC) system with autosampler for both on-line and at-line PAT modes. The methods were applied in UF/DF studies for the comparison of single-use tangential flow filtration (TFF) cassettes to standard reusable cassettes to achieve very high concentration mAb drug substance (DS) in the order of 100-200 g/L. These case studies demonstrated that single-use TFF cassettes are a functionally equivalent, low-cost alternative to standard reusable cassettes, and that the on-line PAT measurement of purity and excipient concentration was comparable to orthogonal offline methods. These PAT applications using an on-line LC system equipped with onboard sample dilution can become a platform system for monitoring of multiple attributes over a wide dynamic range, a potentially valuable tool for biological drug development and manufacturing.

The biopharmaceutical industry is transitioning from currently deployed batch-mode bioprocessing to a highly efficient and agile next generation bioprocessing with the adaptation of continuous bioprocessing, which reduces the capital investment and operational costs. Continuous bioprocessing, aligned with FDA’s quality-by-design (QbD) platform, is designed to develop robust processes to deliver safe and effective drugs. With the deployment of knowledge based operations, product quality can be built into the process to achieve desired critical quality attributes (CQAs) with reduced variability. To facilitate next generation continuous bio-processing, it is essential to embrace a fundamental shift-in-paradigm from “quality-by-testing” to “quality-by-design”, which requires the deployment of process analytical technologies (PAT). With the adaptation of PAT, a systematic approach of process and product understanding and timely process control are feasible. Deployment of PAT tools for real-time monitoring of CQAs and feedback control is critical for continuous bioprocessing. Given the current deficiency in PAT tools to support continuous bioprocessing, we have integrated Agilent 2D-LC with a post-flow-splitter in conjunction with the SegFlow automated sampler to the bioreactors. With this integrated system, we have established a platform for online measurements of titer and CQAs of monoclonal antibodies (mAbs) as well as amino acid concentrations of bioreactor cell culture.

Essential commodities are always on demand by people at a nominal rate during normal operations but due to COVID-19 and various other reasons, there is a sudden rise in demand for essentials goods. Every marketer who was not associated with essential goods tried to reposition their marketing strategy and tried to incorporate essential goods. During COVID-19, many people have turned towards essential goods sector for a living, and because of the rising demand and supply challenge, the government has acted to make essential products available to even the poorest parts of society. Artificial Intelligence (AI) would be essential to the supply chains of the next decade, effortlessly combining long haul to last-mile capabilities with commodity production, availability, and markets. Supply chains facilitated by AI can also help to respond to emergencies and pandemics quicker and faster, saving lives. The present study determines the important key points to be considered for formulating the utility and traceability of Artificial Intelligence in making the availability of essential goods.

The COVID-19 pandemic has propelled a tech-savvy route into the education sector. While it is clear that online courses and tests cannot replace traditional teaching techniques, it is not possible to ignore the role of technology and its widespread use in teaching. The study specifically projects the value of technology for academic assistance of online training and evaluation. Moreover, this pandemic has impacted future teaching and learning in many ways. As a result, educators have been facing many challenges during online sessions like no knowledge about online teaching applications. Collaborations between government, technology firms, and higher education institutions in the public, private, and higher education collaboration modes will solve the problem of technological infrastructure.

In many bacteria and in eukaryotes, replication fork establishment requires the controlled loading of hexameric, ring-shaped helicases around DNA by AAA+ ATPases. How loading factors use ATP to control helicase deposition is poorly understood. Here, we dissect how specific ATPase elements ofE. coliDnaC, an archetypal loader for the bacterial DnaB helicase, play distinct roles in helicase loading and the activation of DNA unwinding. We identify a new element, the arginine-coupler, which regulates the switch-like behavior of DnaC to prevent futile ATPase cycling and maintains loader responsiveness to replication restart systems. Our data help explain how the ATPase cycle of a AAA+-family helicase loader is channeled into productive action on its target; comparative studies indicate elements analogous to the Arg-coupler are present in related, switch-like AAA+ proteins that control replicative helicase loading in eukaryotes, as well as polymerase clamp loading and certain classes of DNA transposases.

Indian corporations provide financial statements in a unique way according to the nation’s necessity. Making uniformity in accounting standards will arise as a huge job. This has fixed by the implementation of IFRS, as IFRS is a popular accounting language for the companies located worldwide. IFRS across the globe is comprehensible and similar. Within the modern globalization, technology in which the Indian financial system has also flourished, adopting IFRS would render Indian business enterprise on an equal footing with other global businesses. This will lead to widening the scope of the Indian market’s foreign financing. To converge with IFRS, India also decided by creating an Ind-AS standard. This study examines the perception of accounting professionals toward the awareness and adoption of IFRS.

Background: Acute decompensated heart failure is a leading cause of hospitalizations in adults older than 65 years. While randomized trials have shown medical therapy to be effective in reducing heart failure hospitalisations, real-world data showing the effectiveness of their implementation and impact on readmissions is scarce. This study aimed to evaluate ACC/AHA directed discharge practices for patients with heart failure with reduced ejection fraction (HFrEF) at a community hospital and its effect on 30-day readmission rates. Methods: This was a retrospective analysis of 868 patients admitted to a community hospital with HFrEF. HFrEF was confirmed for patients based on their most recent Echocardiogram. Data collection points included age, comorbidities (HTN, DM, CKD) and ejection fraction on last Echo. There were 2 primary measured outcomes: medications at discharge and 30-day readmission rates for HFrEF. Discharge medications were subdivided into a combination of either {ACEI/ARB, Beta-blocker (BB) and Spironolactone} or {ACEI/ARB and BB} or {Nitrates/Hydralazine}-all combinations recognised as guideline directed medical therapy (GDMT); or BB alone or ACEI alone. Secondary outcomes included fluid and salt restriction recommendations on discharge and its effect on 30-day readmission rates. Results: After applying exclusion criteria, 320 patients admitted from 2016-2019 for HFrEF were assessed. Using descriptive analyses, it was found that 77% of patients (247/320) were appropriately discharged on GDMT including a combination of {ACEI/ARB, BB and Spironolactone (59/320)}, {ACEI/ARB and BB (173/320)} and {Nitrates/Hydralazine (15/320)}, respectively. The remaining 23% patients were discharged on ACEI or BB alone (73/320). Only 4.38% of patients discharged on ACEI/ARB, BB and Spironolactone were readmitted within 30 days. Similar results were seen across the ACEI/ARB and BB arm (9.38%) and the Nitrates/Hydralazine arm (1.88%). Readmission was significantly higher for patients discharged on a BB alone (27.78%) or ACEI alone (28.57%). Readmission rates for patients recommended fluid and salt restriction versus those who were not, were not statistically significant. Conclusion: While ACC/AHA guidelines in accordance with Target HF, recommend early initiation of GDMT in HFrEF, our study aimed to assess real world implementation of these guidelines at a community hospital. Our study found that while our readmission rates remained higher than the national average (21.9%) in patients discharged on ACEI or BB alone, in agreement with trials, readmission rates were significantly lower in patients discharged in each combination of GDMT. Our study highlights the importance of greater focus on providing high-value care with high compliance with GDMT to reduce readmissions and improve patient outcomes.

Substrate-free and -bound states of Lon reveal distinct operational modes used by the enzyme to process protein substrates., Substrate-bound structures of AAA+ protein translocases reveal a conserved asymmetric spiral staircase architecture wherein a sequential ATP hydrolysis cycle drives hand-over-hand substrate translocation. However, this configuration is unlikely to represent the full conformational landscape of these enzymes, as biochemical studies suggest distinct conformational states depending on the presence or absence of substrate. Here, we used cryo–electron microscopy to determine structures of the Yersinia pestis Lon AAA+ protease in the absence and presence of substrate, uncovering the mechanistic basis for two distinct operational modes. In the absence of substrate, Lon adopts a left-handed, “open” spiral organization with autoinhibited proteolytic active sites. Upon the addition of substrate, Lon undergoes a reorganization to assemble an enzymatically active, right-handed “closed” conformer with active protease sites. These findings define the mechanistic principles underlying the operational plasticity required for processing diverse protein substrates.

In the present scenario, high-speed living people need to understand more suitable and safe devices to improve their quality of life. For acquiring this target there is an advanced financial product known as Mobile Banking. This is fully secured as well as much simpler to be utilized specifically by the senior citizens and children. This research has proposed an innovative model of Mobile Banking and which is empirically tested its adoption in BRICS nation’s customers by acquiring the data via questionnaire amongst 160 BRICS nations’ reversers. In this research, the essential nature of this product will be examined in the situation of the BRICS nation as well as the readiness of the BRICS nations to accept this progress with the use of survey methodology through questionnaires. The results are articulated by using the software named as SPSS and results show that many BRICS nationals who have already been presented as respondents are willing to adopt this product if it is more secure as well as results show that Indians prefer security features over other features whereas other countries prefer convenience features of Mobile Banking.

81 p.- 7 fig.-1 tab.-7 fig. supl.-6 vid., In cells, dedicated AAA+ ATPases deposit hexameric, ring-shaped helicases onto DNA to initiate chromosomal replication. To better understand the mechanisms by which helicase loading can occur, we used cryo-EM to determine sub-4-Å-resolution structures of the E. coli DnaB⋅DnaC helicase⋅loader complex with nucleotide in pre- and post-DNA engagement states. In the absence of DNA, six DnaC protomers latch onto and crack open a DnaB hexamer using an extended N-terminal domain, stabilizing this conformation through nucleotide-dependent ATPase interactions. Upon binding DNA, DnaC hydrolyzes ATP, allowing DnaB to isomerize into a topologically closed, pre-translocation state competent to bind primase. Our data show how DnaC opens the DnaB ring and represses the helicase prior to DNA binding and how DnaC ATPase activity is reciprocally regulated by DnaB and DNA. Comparative analyses reveal how the helicase loading mechanism of DnaC parallels and diverges from homologous AAA+ systems involved in DNA replication and transposition., This work was supported by the NIGMS (to J.M.B., R37-071747), and by the Spanish MINECO/AEI, the ERDF ESF, (to E.A.-P., BFU2017-89143-P and RYC-2015-19059)., Access to eBIC was supported by iNEXT, grant number 653706, funded by the Horizon 2020 program of the European Commission.

Background Carcinoma cervix accounts for 12% of all malignancies worldwide. Pap smear is the most popular screening tool to detect cervical lesion since 1960s. However, this screening method requires an established laboratory a trained cytologist and a repeat visit for its report and further evaluation. In 1982 visual inspection after application of acetic acid on the cervix was reported as an alternative method for screening of cervical cancer. Application of 3–5% acetic acid causes dehydration of cells and surface coagulation of proteins in active cells, giving its white appearance called acetowhite areas. Method We conducted a cross sectional study in the outpatient Department of Obstetrics and Gynaecology, Sir Ganga Ram Hospital from November 2008 to November 2010. The objectives of this study were to evaluate the utility of visual inspection with acetic acid (VIA) for screening of precancerous or cancerous lesion of cervix and to compare it with papanicolaou smear. Results The prevalence of abnormal smear (more than CIN II) amongst women screened with Pap smear in our study was 5.8%. The sensitivity of VIA and Pap smear in our study to diagnose precancerous and invasive disease was comparable 93.1% vs 86.2%. Both screening tests had low specificity VIA 32.2% and Pap smear 60%. Conclusion We suggest that VIA with high sensitivity is comparable to Pap smear can be used as an alternative screening test, especially in low resource settings where the expertise for definite tests is not available. The screen and refer approach for further tests can be used, although there are limitations of this method in terms of low specificity and lack of standardization.

t is estimated that 5-8 million patients present to the emergency department annually for chest pain and a differentiating acute coronary syndrome from non-cardiac chest pain is the primary diagnostic challenge. Serial cardiac troponins are measured to aid the diagnosis. Low-risk patients (TIMI

Summary Through targeted inactivation of the ssrA and smpB genes, we establish that the trans-translation process is necessary for normal growth, adaptation to cellular stress and virulence by the bacterial pathogen Francisella tularensis. The mutant bacteria grow slower, have reduced resistance to heat and cold shocks, and are more sensitive to oxidative stress and sublethal concentrations of antibiotics. Modifications of the tmRNA tag and use of higher-resolution mass spectrometry approaches enabled the identification of a large number of native tmRNA substrates. Of particular significance to understanding the mechanism of trans-translation, we report the discovery of an extended tmRNA tag and extensive ladder-like pattern of endogenous protein-tagging events in F. tularensis that are likely to be a universal feature of tmRNA activity in eubacteria. Furthermore, the structural integrity and the proteolytic function of the tmRNA tag are both crucial for normal growth and virulence of F. tularensis. Significantly, trans-translation mutants of F. tularensis are impaired in replication within macrophages and are avirulent in mouse models of tularemia. By exploiting these attenuated phenotypes, we find that the mutant strains provide effective immune protection in mice against lethal intradermal, intraperitoneal and intranasal challenges with the fully virulent parental strain.

We present proof-of-concept results for the elimination of waterborne bacteria by reactive minerals. We exposed Escherichia coli MG1655 suspended in water to the reactive mineral pyrite (FeS2) at room temperature and ambient light. This slurry eliminates 99.9% of bacteria in fewer than 4 hours. We also exposed Escherichia coli to pyrite leachate (supernatant liquid from slurry after 24 hours), which eliminates 99.99% of bacteria over the same time-scale. Unlike SOlar water DISinfection (SODIS), our results do not depend on the presence of ultraviolet (UV) light. We confirmed this by testing proposed SODIS additive and known photo-catalyst anatase (TiO2) for antibacterial properties and found that, in contrast to pyrite, it does not eliminate E. coli under our experimental conditions. Previous investigations of naturally antibiotic minerals have focused on the medical applications of antibiotic clays, and thus have not been conducted under experimental conditions resembling those found in water purification. In our examination of the relevant literature, we have not found previously reported evidence for the use of reactive minerals in water sanitization. The results from this proof-of-concept experiment may have important implications for future directions in household water purification research.

The AAA+ Lon protease is conserved from bacteria to humans, performs crucial roles in protein homeostasis, and is implicated in bacterial pathogenesis and human disease. We investigated how Lon selectively degrades specific substrates among a diverse array of potential targets. We report the discovery of HspQ as a new Lon substrate, unique specificity-enhancing factor, and potent allosteric activator. Lon recognizes HspQ via a C-terminal degron, whose precise presentation, in synergy with multipartite contacts with the native core of HspQ, is required for allosteric Lon activation. Productive HspQ-Lon engagement enhances degradation of multiple new and known Lon substrates. Our studies reveal the existence and simultaneous utilization of two distinct substrate recognition sites on Lon, an HspQ binding site and an HspQ-modulated allosteric site. Our investigations unveil an unprecedented regulatory use of an evolutionarily conserved heat shock protein and present a distinctive mechanism for how Lon protease achieves temporally enhanced substrate selectivity.