Dual Inhibition of Cathepsin L and 3CL-Pro by GC-376 Constrains SARS Cov2 Infection Including Omicron Variant

Recurrent waves of SARS CoV2 infections remain a major global health concern. Emergence of highly infectious variants with reduced sensitivity to neutralization by vaccines and monoclonal antibodies (mAb) necessitates a deeper understanding of factors involved in SARS CoV2 infections and identification of drug candidates to halt infection. Here, we determined the primacy of endosomal protease cathepsin-L in mediating SARS CoV2 entry and screened a library of well-annotated bioactive compounds for potent cathepsin-L inhibitory activity. Whilst the potent cathepsin-L inhibitors were capable of inhibiting SARS CoV2 entry and cytopathic effect (CPE) in less susceptible cell lines such as human ACE2 expressing 293T cells, these drugs failed to inhibit SARS CoV2 in highly susceptible cell lines such as human TMPRSS2 or human-ACE2-TMPRSS2 overexpressing Vero E6 cells. Only drugs with dual inhibitory effect on both host cathepsin-L and virus 3CL-Protease enzymes such as Z-FA-FMK and GC-376 were capable of inhibiting prototypic (USA-WA1/2020, Lineage A) SARS CoV2 induced CPE in highly susceptible cell lines. Moreover, these drugs inhibited delta (Lineage-B.1.617.2) and omicron (Lineage-B.1.1.529) infection with equal potency showing that the newer mutations harbored in these variants did not affect the mechanism of action of these drugs such as cathepsin-L or 3CL-Pro inhibition. Moreover, our early evidence that 3CL-Pro inhibition can effectively inhibit omicron-induced CPE in highly susceptible cell lines suggests that the recently FDA-approved oral drug, a 3CL-Pro inhibitor which is a combination of nirmatrelvir/ritonavir (Paxlovid) could be effective against omicron variant which shows reduced sensitivity to vaccines and mAb. ImportanceWe report that cathepsin-L and 3CL-Pro as major targets for designing antivirals against SARS CoV2. Dual inhibition of cathepsin-L and 3CL-Pro by GC-376 renders it effective in inhibiting SARS CoV2-induced cytopathic effect in highly susceptible cell lines. Moreover, this candidate drug is equally effective against prototypic SARS CoV2 lineage A and emerging variants such as delta and omicron which show reduced sensitivity to vaccines and monoclonal antibodies. Given the recent wave of SARS CoV2 omicron variant infection around the world, and 3CL-Pro inhibitor nirmatrelvir is one of the components of the FDA-approved Paxlovid, our findings are timely, important and should be of broad interest.

A Simulated Graphical Interface for Integrating Patient-Generated Health Data From Smartwatches With Electronic Health Records: Usability Study.

BACKGROUND: Wearable technology, such as smartwatches, can capture valuable patient-generated data and help inform patient care. Electronic health records provide logical and practical platforms for including such data, but it is necessary to evaluate the way the data are presented and visualized. OBJECTIVE: The aim of this study is to evaluate a graphical interface that displays patients' health data from smartwatches, mimicking the integration within the environment of electronic health records. METHODS: A total of 12 health care professionals evaluated a simulated interface using a usability scale questionnaire, testing the clarity of the interface, colors, usefulness of information, navigation, and readability of text. RESULTS: The interface was positively received, with 14 out of the 16 questions generating a score of 5 or greater among at least 75% of participants (9/12). On an 8-point Likert scale, the highest rated features of the interface were quick turnaround times (mean score 7.1), readability of the text (mean score 6.8), and use of terminology/abbreviations (mean score 6.75). CONCLUSIONS: Collaborating with health care professionals to develop and refine a graphical interface for visualizing patients' health data from smartwatches revealed that the key elements of the interface were acceptable. The implementation of such data from smartwatches and other mobile devices within electronic health records should consider the opinions of key stakeholders as the development of this platform progresses.

Secondary care provider attitudes towards patient generated health data from smartwatches.

Wearable devices, like smartwatches, are increasingly used for tracking physical activity, community mobility, and monitoring symptoms. Data generated from smartwatches (PGHD_SW) is a form of patient-generated health data, which can benefit providers by supplying frequent temporal information about patients. The goal of this study was to understand providers' perceptions towards PGHD_SW adoption and its integration with electronic medical records. In-depth, semi-structured qualitative interviews were conducted with 12 providers from internal medicine, family medicine, geriatric medicine, nursing, surgery, rehabilitation, and anesthesiology. Diffusion of Innovations was used as a framework to develop questions and guide data analysis. The constant comparative method was utilized to formulate salient themes from the interviews. Four main themes emerged: (1) PGHD_SW is perceived as a relative advantage; (2) data are viewed as compatible with current practices; (3) barriers to overcome to effectively use PGHD_SW; (4) assessments from viewing sample data. Overall, PGHD_SW was valued because it enabled access to information about patients that were traditionally unattainable. It also can initiate discussions between patients and providers. Providers consider PGHD_SW important, but data preferences varied by specialty. The successful adoption of PGHD_SW will depend on tailoring data, frequencies of reports, and visualization preferences to correspond with the demands of providers.

Quantification of uncertainties in conventional plan evaluation methods in Intensity Modulated Radiation Therapy.

PURPOSE: In Intensity Modulated Radiation Therapy (IMRT) dose distributions tend to be more complex and heterogeneous because of the modulated fluences in each beamlet of every single beam. These dose-volume (DV) parameters derived from the dose volume histogram (DVH) are physical quantities, thought to correlate with the biological response of the tissues. The aim of this study was to quantify the uncertainty of physical dose metrics to predict clinical outcomes of radiotherapy. METHODS: The radiobiological estimates such as tumor control probability (TCP) and Normal Tissue Complication Probability (NTCP) were made for a cohort of 40 cancer patients (10 brain;19 head & neck;11 cervix) using the DV parameters. Statistical analysis was performed to determine the correlation of physical plan quality indicators with radiobiological estimates. RESULTS: The correlation between conformity index (CI) and TCP was found to be good and the dosimetric parameters for optic nerves, optic chiasm, brain stem, normal brain and parotids correlated well with the NTCP estimates. A follow up study (median duration 18 months) was also performed. There was no grade 3 or 4 normal tissue complications observed. Local tumor control was found to be higher in brain (90%) and pelvic cases (95%), whereas a decline of 70% was noted with head & neck cancer cases. CONCLUSIONS: The equivalent uniform dose (EUD) concept of radiobiological model used in the software determines TCP and NTCP values which can predict outcomes precisely using DV data in the voxel level. The uncertainty of using physical dose metrics for plan evaluation is quantified with the statistical analysis. Radiobiological evaluation is helpful in ranking the rival treatment plans also.

Cerebrospinal fluid profile of amyloid ß42 (Aß42), hTau and ubiquitin in North Indian Alzheimer's disease patients.

Alzheimer's disease (AD) is the most common form of dementia, and is characterized by the degeneration of neurons and their synapses, and a higher number of amyloid plaques and neurofibrillary tangles (NFTs) compared with that found in non-demented individuals. Amyloid-ß-peptides (Aß) are major components of amyloid plaques in AD brain whereas NFTs are composed of Tau and associated with ubiquitin. The aim of the present study was to analyze the levels of Aß42, hTau (total Tau) and ubiquitin in CSF of North Indian population. CSF Aß42, Tau and ubiquitin were measured in CSF of AD patients as well as controls using ELISA assays. Here we report low Aß42 levels in AD patients (324.24±76.38pg/ml) as compared to those in non-AD (NAD) (668.34±43.13pg/ml), neurological controls (NCs) (727.28±46.49pg/ml) and healthy controls (HCs) (976.47±124.46pg/ml). In contrast, hTau and ubiquitin levels were significantly high (568.65±48.89pg/ml and 36.82±4.34ng/ml, respectively) in AD patients compared to those in NAD, NC and HC. The hTau levels were 267.37±36.64pg/ml, 167.34±44.27pg/ml and 107.62±24.27pg/ml in NAD, NC and HC, respectively. Similarly, ubiquitin levels were 23.57±2.32ng/ml, 19.76±3.64ng/ml and 13.24±4.56ng/ml in NAD, NC and HC, respectively. In conclusion, low Aß42 and high Tau-ubiquitin levels were found in North Indian AD patients.

Efficacy of SPECT over planar bone scan in the diagnosis of solitary vertebral lesions in patients with low back pain.

BACKGROUND: The purpose of our study has been to evaluate the efficacy of single photon emission computed tomography (SPECT) over planar bone scan in identifying solitary vertebral lesions in patients with low backache and its ability to differentiate various pathologies according to the uptake pattern. MATERIALS AND METHODS: The study included twenty patients out of whom six patients presented with known carcinoma and fourteen patients with low back pain. SPECT was done in all following planar skeletal survey. Benign and malignant lesions were identified according to the uptake pattern in vertebral elements, based on Gary F. Gates observations. Final diagnosis was obtained by means of biopsy or correlation with radiograph or computed tomography (CT) or magnetic resonance imaging (MRI), and / or follow up. RESULTS: SPECT detected additional 30% of solitary vertebral lesions that were obscured on planar scan. Seven out of twenty were localized in anterior vertebral body and were diagnosed as benign ostophytes in six and osteoma in one substantiating the previous observations. Out of six cases of known carcinoma, three were having solitary metastases and showed posterior vertebral body uptake with pedicle involvement. SPECT could localize specific lesions as source of pain in eleven patients with low back pain (78%) and identified various etiologies including benign tumors (osteoid osteoma and osteoma), facet arthritis, discitis, transverse process fractures and spondylolysis. CONCLUSION: Our study highlighted the higher diagnostic value of SPECT over planar skeletal scintigraphy in localizing solitary vertebral lesions in low backache patients. Based on SPECT pattern, malignant and benign lesions could be differentiated in the given clinical context.

Detection of lead in vegetables with new chromogenic reagent by spectrophotometry.

A new simple, rapid selective and highly sensitive chromogenic reagent dibromo-p-methyl-carboxyazo (DBMCA) was synthesized and studied in detail for the spectrophotometric detection of lead. In 0.25 M phosphoric acid medium, which greatly increases the selectivity, Lead reacts with DBMCA to form a 1:2 blue complex having a sensitivity absorption peak at 646 nm. Under optimal conditions, Beer's Law is obeyed over the range from 0.09 to 0.8 microg mL(-1) Pb (II) and the apparent molar absorptivity is 1.03 x 10(5) mL(-1) cm(-1). The detection limit and the variation coefficient were found to be 2.12 microg mL(-1) and 1.0% respectively. It is found that, except for Ca (II) and Ba (II) all foreign ions studied do not interfere with detection. The interference caused by Ca (II) and Ba (II) can be easily eliminated by prior extraction with potassiumiodide-methylisobutylketone. The proposed method has been applied successfully for to the detection of Lead in vegetable leaves with good results.