Pharmacological Evaluations of Select Herbal Extracts on TLR7/8-induced Cytokine and Chemokine Production in Macrophage-like Cells (preprint)

Inflammation is an innate immune response triggered by harmful stimuli, such as pathogens, tissue injury, or toxins. The purpose is to eliminate the source of infection and initiate the healing process. However, an excessive acute inflammatory response can lead to severe and life-threatening complications, as seen during recent pandemics. In the context of viral infections, the activation of the TLR7/8 signaling pathway has been implicated in excessive cytokine secretion. In this study, we aimed to replicate the perturbed inflammatory environment resulting from the activation of the TLR7/8 specific agonists, imiquimod, and resiquimod. We utilized macrophage-like cells, as macrophages are the first responders during infections and secrete key pro-inflammatory cytokines (TNF-, IL-6, and IL-1{beta}) to recruit immune cells to the site of infection. Herbal medicines have been traditionally used for centuries to enhance respiratory immune function. In the present study, we employed a prophylactic approach, where macrophage-like THP1 cells, differentiated with PMA, were pre-treated with select herbal extracts/formulations prior to TLR7/8 activation in the presence of agonists. Several medicinal plants and formulations known for their therapeutic potential in respiratory ailments were investigated, including Withania somnifera, Tinospora cordifolia, Glycyrrhiza glabra, and AYUSH-64, an herbal formulation. The gene expression and corresponding secreted levels of various inflammatory mediators were measured using RT-PCR and ELISA methods, respectively, in treated and untreated differentiated THP1 cells induced with TLR7/8 agonists. Comparatively, the gene expression of inflammatory markers was significantly higher in resiquimod-induced cells than in imiquimod-treated cells. Notably, Withania somnifera demonstrated pronounced prophylactic efficacy compared to other herbs/formulations, as evidenced by reduction in expression of majority of investigated inflammatory marker genes.

An in silico investigation of the phytomolecules present in Piper longum roots as a potential treatment for SARS-CoV-2 (preprint)

The epidemic of COVID-19 brought on by SARS-CoV-2 (coronavirus related to SARS) as well as its variants is still a life-threatening and critical risk to public health around the world. Due to the COVID-19 virus's propensity for mutation, the pandemic is still spreading havoc compared to SARS-CoV-2. Several phytochemicals are now undergoing in silico analysis for their anti-SARS-CoV-2 efficacy. Scientists all over the world are on the lookout for novel lead phytochemicals that can effectively block SARS-CoV-2 entrance and infection in host cells. Piper longum phytomolecules suggest that it may be more effective to use natural remedies and herbal extracts to improve immunity and fight the infection. In this current in silico study of different Piper longum phytochemicals like sesamin, piperine, gamma-sitosterol, and epizonarene, Utilizing Lipinski's rule, study the drugability guidelines and oral bioavailability of these compounds. Four putative SARS-CoV-2 therapeutic targets were explored using the top four ligands in a comparative study. Our finding shows that sesamin, piperine, gamma-sitostenone, and epizonarene all displayed binding affinity values of -8.9, -7.8, -7.4, and − 7.2 kcal/mol, respectively. Sesamin was found to have a greater negative binding energy toward the SARS-CoV-2 targets, which is a significant finding. We concluded that sesamin may be effective as a treatment for SARS-CoV-2.

Coronavirus disease 2019 and mechanical circulatory support devices: A comprehensive review.

Coronavirus disease (COVID-19) can cause circulatory shock refractory to medical therapy. Such patients can be managed with mechanical circulatory support (MCS) devices like IABP, Impella, VA ECMO, and Left Ventricular Assist Devices (LVADs). Moreover, patients on long-term durable LVADs are a special population having increased susceptibility and mortality to COVID-19 infection. In this narrative review, we searched PubMed and Medline for studies on COVID-19 patients on short-term MCS devices. We found 36 papers with 110 patients who met our review criteria, including 89 LVAD patients and 21 COVID-19 patients who needed MCS device therapy. These studies were used to extract patient demographics, clinical presentation, MCS device details, management, and outcomes. Mean age of patients with COVID-19 infection on LVADs was 60, 73% were male, and HeartMate 3 was the most common device (53%). Most patients (77.5%) needed hospitalization, and mortality was 23.6%. Among the 21 reported cases of critically ill COVID-19 patients who required MCS, the mean age was 49.8 years, 52% were women, and the most common MCS device used was VA ECMO (62%) in conjunction with an Impella for LV venting. Comorbidities were not present in 43%, but 71% had abnormal ventricular function on echocardiography. MCS is a viable option for managing severe COVID-19 infection with shock, with many reported cases of favorable outcomes.

Correlating the differences in the receptor binding domain of SARS-CoV-2 spike variants on their interactions with human ACE2 receptor.

Spike glycoprotein of SARS-CoV-2 variants plays a critical role in infection and transmission through its interaction with human angiotensin converting enzyme 2 (hACE2) receptors. Prior findings using molecular docking and biomolecular studies reported varied findings on the difference in the interactions among the spike variants with the hACE2 receptors. Hence, it is a prerequisite to understand these interactions in a more precise manner. To this end, firstly, we performed ELISA with trimeric spike glycoproteins of SARS-CoV-2 variants including Wuhan Hu-1(Wild), Delta, C.1.2 and Omicron. Further, to study the interactions in a more specific manner by mimicking the natural infection, we developed hACE2 receptors expressing HEK-293T cell line, evaluated their binding efficiencies and competitive binding of spike variants with D614G spike pseudotyped virus. In line with the existing findings, we observed that Omicron had higher binding efficiency compared to Delta in both ELISA and Cellular models. Intriguingly, we found that cellular models could differentiate the subtle differences between the closely related C.1.2 and Delta in their binding to hACE2 receptors. Our study using the cellular model provides a precise method to evaluate the binding interactions between spike sub-lineages to hACE2 receptors.

Intravenous BCAA infusion does not lead to a sustained recovery from overt HE in ACLF – An open label randomized clinical trial

Background –Hepatic encephalopathy(HE) in acute-on-chronic liver failure(ACLF) is associated with significant morbidity and mortality. We conducted a prospective, randomized controlled clinical trial to study efficacy of intravenous branched chain amino acids(IV-BCAA) with lactulose versus lactulose alone for improvement in HE at 24h, day 3 & day 7. Primary outcome was improvement in encephalopathy by ≥ 1 grade at 72 hours. Patients and Methods –EASL defined ACLF patients with overt HE were assessed and randomized into experimental arm (IV-BCAA - 500mL/day for 3 days + Lactulose;n=39) and comparator arm (Lactulose alone;n=37). Six patients developed COVID-19 after randomization & were excluded (4-experimental arm & 2-comparator arm). Results –Of 222 screened patients, 70 (35 in each arm) were included in analysis. Baseline characteristics including HE grade (2.9 ± 0.7 vs 2.8 ± 0.7;P = 0.86) and CLIF-C ACLF score (54.2 ± 5.6 vs 54.8 ± 5.7;P = 0.65) were similar. Overall survival was 40% at 28 days (48.5% vs 31.4%;P=0.14). Improvement in HESA by ≥1 grade at 24h occurred in 14 patients (40%) in BCAA arm and 6 patients (17.1%) in control group (P=0.03) which translated to shorter ICU stay. Median change in HESA at 24h was more in BCAA arm than control arm(P=0.006) which was not sustained at day 3 or 7. Ammonia levels did not correlate with grade of HE (Spearman's correlation coefficient(ρ) = - 0.0843;P=0.29). Conclusion Intravenous BCAA does not lead to a sustained improvement in HE grade in ACLF. Trial registration no NCT04238416 (clinicaltrials.gov)

A review of the antiviral activity of cationic antimicrobial peptides.

Viral epidemics are occurring frequently, and the COVID-19 viral pandemic has resulted in at least 6.5 million deaths worldwide. Although antiviral therapeutics are available, these may not have sufficient effect. The emergence of resistant or novel viruses requires new therapies. Cationic antimicrobial peptides are agents of the innate immune system that may offer a promising solution to viral infections. These peptides are gaining attention as possible therapies for viral infections or for use as prophylactic agents to prevent viral spread. This narrative review examines antiviral peptides, their structural features, and mechanism of activity. A total of 156 cationic antiviral peptides were examined for information of their mechanism of action against both enveloped and non-enveloped viruses. Antiviral peptides can be isolated from various natural sources or can be generated synthetically. The latter tend to be more specific and effective and can be made to have a broad spectrum of activity with minimal side effects. Their unique properties of being positively charged and amphipathic enable their main mode of action which is to target and disrupt viral lipid envelopes, thereby inhibiting viral entry and replication. This review offers a comprehensive summary of the current understanding of antiviral peptides, which could potentially aid in the design and creation of novel antiviral medications.

Venous Thromboembolism and COVID-19-an Epidemiological Perspective.

The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was declared as pandemic by World Health Organization (WHO) in March 2020. The outbreak has caused 5,232,562 deaths worldwide until December 3rd, 2021. Though primarily affecting the respiratory system, involvement of other organ systems have been reported in severe disease. Venous thromboembolism (VTE) has been recognized as an important complication. Previous studies have reported the prevalence of VTE in intensive care unit (ICU) patients between 7 and 85% and in non-ICU patients between 0 and 19%. COVID-19 patients that are at high risk for VTE are also at increased risk for bleeding. In such cases, anticoagulation may potentially be harmful. Thereby, it is important to understand the risk factors for VTE predisposition in the COVID-19 patients, timing of VTE, and the rate of occurrence of VTE in hospitalized patients post-discharge. Comparison of the rate of occurrence of VTE in COVID-19 patients with the non-COVID-19 patients with similar disease severity is required to truly interpret the reportedly high rates of VTE in COVID-19 patients. Several pathophysiological mechanisms have been reported for the development of VTE in COVID-19. Autopsy-based studies have contributed to the existing knowledge. d-dimer, presently, seems to be the most suitable investigation for risk-identification of VTE supported by Doppler studies and overall clinical context. Further, prospective studies and clinical trials are essentially required to fill the gaps in evidence for occurrence, risk prediction and management of VTE in COVID-19 patients.

FINANCING INDIA'S URBAN INFRASTRUCTURE: ISSUES AND CHALLENGES

Indian economy is expected to be one of the top performers in the world economy in the post covid era. While all the major economies of the world are finding it difficult to come back to pre-covid level, the Indian economy is on the path of a speedy recovery. With significant urbanization taking place, India will need to make a significant investment in its urban infrastructure. It is expected that $840 billion of investments is required in urban infrastructure over the next 15 years. This private player will have a significant role to play. This article attempts to identify the significant constraints in financing such a massive infrastructure needs of India.

An Analysis of Profitability and Key Hurdles for the Housing Finance Sector in India

There is a fear of recession worldwide as is evident from the decline in the economic activities in all major economies of the world. As per the prediction of the IMF, the Indian economy looks somewhat better placed compared to other major economies for the current financial year. The real estate sector has always been a significant contributor to the process of growth of India and depends on the soundness of the Housing Finance sector to a great extent. The study tries to examine the profitability of the housing finance sector as well as identifies significant hurdles, especially in the post Covid period when the fear of global recession looms large. Three leading housing finance companies based on market capitalization are taken as the sample companies and one-way Anova is used as a significant tool of analysis. The study points that the favourable time ahead for housing finance sector in near future.

Low dose (ICMR) hydroxychloroquine prophylaxis for COVID-19 fails - a prospective study

Since the COVID-19 pandemic, the world began a frantic search for possible prophylactic options. We conducted a study to assess the role of hydroxychloroquine for COVID-19 prophylaxis in health-care workers. The study was a prospective cohort with four arms (high, medium, low dose, and control) of HCQ prophylaxis. Participants were grouped as per their opting for any one arm on a voluntary basis as per institute policy. The outcomes studied were COVID-19 positivity by RT-PCR and its severity assessed by WHO COVID-19 severity scale. Total 486 participants were enrolled, of which 29 (6%) opted for low dose, 2 (<1%) medium dose, and none for high dose HCQ while 455 (93.6%) were in the control arm. Of the 164 participants who underwent RT-PCR, 96 (58.2%) tested positive. Out of these 96 positive cases, 79 [82.3%] were ambulatory and were managed conservatively at home. Only 17.7% participants, all from the control group, required hospitalization with the mild-moderate disease. None of the participants had severe disease, COVID-related complications, ICU stay, or death. The difference in the outcome was statistically insignificant (p value >0.05). This single-centre study demonstrated that HCQ prophylaxis in healthcare workers does not cause a significant reduction in COVID-19 as well as mitigating its severity in those infected. At present, most of the trials have not shown any benefit. Though COVID-19 vaccines have reduced the need for prophylaxis, the search for a safe and reasonable chemoprophylaxis should continue until a large population of individuals gets vaccinated, especially in underdeveloped countries.

Flavan-based phytoconstituents inhibit Mpro, a SARS-COV-2 molecular target, in silico.

A well-validated in-silico approach can provide promising drug candidates for the treatment of the ongoing CoVID19 pandemic. In this study, we have screened 32 phytochemical constituents (PCCs) with Mpro binding site (PDB:6W63) based on which we identified three possible candidates that are likely to be effective against CoVID19-viz., licoleafol (binding energy: -8.1 kcal/mol), epicatechin gallate (-8.5 kcal/mol) and silibinin (-8.4 kcal/mol) that result in higher binding affinity than the known inhibitor, X77 (-7.7 kcal/mol). Molecular dynamics (MD) simulations of PCCs-Mpro complex confirmed molecular docking results with high structural and dynamical stability. The selected compounds were found to exhibit low mean squared displacements (licoleafol: 2.25 ± 0.43 Å, epicatechin gallate: 1.93 ± 0.35 Å, and silibinin: 1.39 ± 0.19 Å) and overall low fluctuations of the binding complexes (root mean squared fluctuations below 2 Å). Visualization of the MD trajectories and structural analyses revealed that they remain confined to the initial binding region, with mean fluctuations lower than 3 Å. To access the collective motion of the atoms, we performed principal component analysis demonstrating that the first 10 principal components are the major contributors (approximate contribution of 80%) and are responsible for the overall PCCs motion. Considering that the three selected PCCs share the same flavan backbone and exhibit antiviral activity against hepatitis C, we opine that licoleafol, epi-catechin gallate, and silibinin can be promising anti-CoVID19 drug candidates.Communicated by Ramaswamy H. Sarma.

Redefining Lobe-Wise Ground-Glass Opacity in COVID-19 Through Deep Learning and its Correlation With Biochemical Parameters.

During COVID-19 pandemic qRT-PCR, CT scans and biochemical parameters were studied to understand the patients' physiological changes and disease progression. There is a lack of clear understanding of the correlation of lung inflammation with biochemical parameters available. Among the 1136 patients studied, C-reactive-protein (CRP) is the most critical parameter for classifying symptomatic and asymptomatic groups. Elevated CRP is corroborated with increased D-dimer, Gamma-glutamyl-transferase (GGT), and urea levels in COVID-19 patients. To overcome the limitations of manual chest CT scoring system, we segmented the lungs and detected ground-glass-opacity (GGO) in specific lobes from 2D CT images by 2D U-Net-based deep learning (DL) approach. Our method shows accuracy, compared to the manual method (  âˆ¼ 80%), which is subjected to the radiologist's experience. We determined a positive correlation of GGO in the right upper-middle (0.34) and lower (0.26) lobe with D-dimer. However, a modest correlation was observed with CRP, ferritin and other studied parameters. The final Dice Coefficient (or the F1 score) and Intersection-Over-Union for testing accuracy are 95.44% and 91.95%, respectively. This study can help reduce the burden and manual bias besides increasing the accuracy of GGO scoring. Further study on geographically diverse large populations may help to understand the association of the biochemical parameters and pattern of GGO in lung lobes with different SARS-CoV-2 Variants of Concern's disease pathogenesis in these populations.

Use of hydroxychloroquine and azithromycin combination to treat the COVID-19 infection.

Coronavirus disease 2019 (COVID-19) infection is unequivocally the worst crisis in recent decades, which is caused by a severe acute respiratory virus 2. Currently, there is no effective therapy for the COVID-19 infection. Different countries have different guidelines for treating COVID-19 in the absence of an approved therapy for COVID-19. Therefore, there is an imminent need to identify effective treatments, and several clinical trials have been conducted worldwide. Both hydroxychloroquine [HCQS], chloroquine, and azithromycin (AZ) have been widely used for management based on in vitro studies favoring antiviral effects against the COVID-19 virus. However, there is evidence both in favor and against the use of hydroxychloroquine and azithromycin (HCQS+AZ) combination therapy to manage the COVID-19 infection. The combination of hydroxychloroquine and azithromycin was significantly associated with increased adverse events. However, the inference of these findings was from observational studies. Therefore, large randomized trials are imperative to show the future path for the use of HCQS+AZ combination therapy. However, owing to the ban on HCQS use in COVID-19, this may no longer be essential. This review is on the pharmacology, trials, regimens, and side effects of hydroxychloroquine and azithromycin combination therapy.

Bispecific antibodies and its applications: a novel approach for targeting SARS-Cov-2.

The COVID-19 pandemic remains a severe global threat, with the world engulfed in the struggle against the disease's second or third waves, which are approaching frightening proportions in terms of cases and mortality in many nations. Despite the critical need for effective therapy, there is still uncertainty about the optimal practices for treating COVID-19 with various pharmaceutical approaches. This being third year, global immunity and eradication of SARS-CoV-2 is currently seems to be out of reach. Efforts to produce safe and effective vaccinations have shown promise, and progress is being made. Additional therapeutic modalities, as well as vaccine testing in children, are required for prophylaxis and treatment of high-risk individuals. As a result, neutralising antibodies and other comparable therapeutic options offer a lot of promise as immediate and direct antiviral medications. Bispecific antibodies offer a lot of potential in COVID-19 treatment because of their qualities including stability, small size and ease of manufacture. These can be used to control the virus's infection of the lungs because they are available in an inhalational form. To combat the COVID-19 pandemic, innovative approaches with effective nanobodies, high-expression yield and acceptable costs may be required.

A cross-sectional study to determine the psychological distress among pulmonary tuberculosis patients during COVID-19 pandemic.

COVID-19 pandemic had adversely affected the services of the National Tuberculosis (TB) Elimination Programme, resulting in psychological distress among pulmonary tuberculosis patients (PTB). This cross-sectional, hospital-based study included 361 PTB patients. Three pre-defined questionnaires were used for the analysis, a questionnaire to evaluate anxiety related to COVID-19, a patient health questionnaire (PHQ-9) for depression, and a fear of COVID-19 scale (FCV-19S) questionnaire. Among 361 PTB patients, 13% (n=47) had COVID-19 infection. Out of the total patients, 69% (n=250) were DR-TB (drug resistance-tuberculosis) cases. Proportion of anxiety, fear and depression due to COVID-19 was found in 49% (n=177), 23% (n=83), 67% (n=247) respectively. Delay in the initiation of anti-tubercular treatment was found in 58% (n=210) of the cases, among which the majority, i.e., 69% (n=172, p=0.011), were DR-TB. This pandemic has led to a sudden step-down of PTB. Trend analysis of the psychological distress showed a peak following the COVID-19 pandemic. Most DR-TB patients had delayed initiation of the anti-tubercular treatment during the pandemic. The preponderance of the younger age group was seen in the pulmonary tuberculosis patients, and a majority of them had DR-TB. Depression was the predominant psychological distress among the study subjects during the pandemic.

Caregiver Burden in Caregivers of Stroke Survivors: A Hospital-Based Study.

Background: A principal caregiver (CG) is directly affected by the patient's health problems leading to CG strain. In the present study, we evaluated the different factors related to increased CG burden in stroke survivors and assessed the relationship between patient's personal and clinical characteristics and CG's stress. Material and Methods: In this prospective, follow-up study, a total of 141 principle CGs of 164 First-ever stroke (FES) survivors were subjected to the Caregivers Strain Index (CSI) and Oberst Caregiving Burden Scale (OCBS) at 30 days (n = 141), 90 days (n = 129), and 6 months (n = 119) after informed consent. Patients were subjected to modified Rankin Scale (mRS) and Barthel index score assessment at the end of 30 days. Results: The mean age of CG was 49.8 ± 21.0 years, approximately 20 years lesser than that of the patients. 102 (72.34%) CGs were females. Urinary incontinence (p < 0.006) morbidity at 30 days, mRS (p = 0.004), and moderate to the severe neurological deficit on admission (p = 0.003) were the patient factors in FES cases leading to significant CGs stress. CG factors responsible for major stress were long caregiving hours (P < 0.001), anxiety (P < 0.001), disturbed night sleep (P < 0.001), financial stress (P < 0.001), younger age (P = 0.002), and CGs being daughters-in-law (P = 0.039). Conclusion: CG burden increases with increased severity of stroke. Integrated stroke rehabilitation services should also address CGs issues along with patients.

Agronomic biofortification of food crops: An emerging opportunity for global food and nutritional security.

Fortification of food with mineral micronutrients and micronutrient supplementation occupied the center stage during the two-year-long Corona Pandemic, highlighting the urgent need to focus on micronutrition. Focus has also been intensified on the biofortification (natural assimilation) of mineral micronutrients into food crops using various techniques like agronomic, genetic, or transgenic. Agronomic biofortification is a time-tested method and has been found useful in the fortification of several nutrients in several crops, yet the nutrient use and uptake efficiency of crops has been noted to vary due to different growing conditions like soil type, crop management, fertilizer type, etc. Agronomic biofortification can be an important tool in achieving nutritional security and its importance has recently increased because of climate change related issues, and pandemics such as COVID-19. The introduction of high specialty fertilizers like nano-fertilizers, chelated fertilizers, and water-soluble fertilizers that have high nutrient uptake efficiency and better nutrient translocation to the consumable parts of a crop plant has further improved the effectiveness of agronomic biofortification. Several new agronomic biofortification techniques like nutripriming, foliar application, soilless activation, and mechanized application techniques have further increased the relevance of agronomic biofortification. These new technological advances, along with an increased realization of mineral micronutrient nutrition have reinforced the relevance of agronomic biofortification for global food and nutritional security. The review highlights the advances made in the field of agronomic biofortification via the improved new fertilizer forms, and the emerging techniques that achieve better micronutrient use efficiency of crop plants.