Role of auto-antibodies in the mechanisms of dengue pathogenesis and its progression: a comprehensive review.

A complex interaction among virulence factors, host-genes and host immune system is considered to be responsible for dengue virus (DENV) infection and disease progression. Generation of auto-antibodies during DENV infection is a major phenomenon that plays a role in the pathophysiology of dengue hemorrhagic fever and dengue shock syndrome. Hemostasis, thrombocytopenia, hepatic endothelial dysfunction, and autoimmune blistering skin disease (pemphigus) are different clinical manifestations of dengue pathogenesis; produced due to the molecular mimicry of DENV proteins with self-antigens like coagulation factors, platelets and endothelial cell proteins. This review elaborately describes the current advancements in auto-antibody-mediated immunopathogenesis which inhibits coagulation cascade and promotes hyperfibrinolysis. Auto-antibodies like anti-endothelial cell antibodies-mediated hepatic inflammation during severe DENV infection have also been discussed. Overall, this comprehensive review provides insight to target auto-antibodies that may act as potential biomarkers for disease severity, and a ground for the development of therapeutic strategy against DENV.

Homeopathic preparation of Allium sativum abrogates OxLDL mediated atherogenic events in macrophages: An invitro and in silico approach.

BACKGROUND: Oxidized LDL (OxLDL), the key player in atherogenesis modulates endothelial dysfunction, initiates monocyte recruitment, accentuates foam cell formation, and flares up inflammatory and apoptotic events. Even though homeopathic preparation of Allium sativum has been proved to be an anti-inflammatory, anti-apoptotic and anti-atherogenic agent, its mechanism of action on abrogating OxLDL mediated foam cell formation is yet to be explored. OBJECTIVE: This study was designed to bring out the role of homeopathic preparation of Allium sativum in curbing OxLDL mediated cellular inflammation in IC-21 cells exposed with OxLDL. MATERIALS AND METHODS: OxLDL was used to induce oxidative damage in the IC-21 macrophage cells. Assessment of inflammatory cytokines, localization of NFκB, detection of apoptosis and the in silico analysis were performed in this study. RESULTS: The current study portrays the efficacy of homeopathy medicine as an anti-inflammatory agent, in reducing the levels of inflammatory cytokines and its mRNA expression, suppressing the activity of NFκB and preventing apoptosis in OxLDL treated IC-21 cells. CONCLUSION: To conclude, homeopathic preparation of Allium sativum 6C and 30C potencies are capable of controlling the transcriptional activity of NFκB and apoptosis in IC-21 cells exposed to OxLDL. These results implicate that Allium sativum homeopathic drug can be used as anti-inflammatory agent in reducing atherogenic events as it is capable of preventing OxLDL-mediated injury to macrophages.

Arsenic album 30C exhibits crystalline nano structure of arsenic trioxide and modulates innate immune markers in murine macrophage cell lines.

Macrophages are associated with innate immune response and M1-polarized macrophages exhibit pro-inflammatory functions. Nanoparticles of natural or synthetic compounds are potential triggers of innate immunity. As2O3 is the major component of the homeopathic drug, Arsenic album 30C.This has been claimed to have immune-boosting activities, however, has not been validated experimentally. Here we elucidated the underlying mechanism of Ars. alb 30C-mediated immune priming in murine macrophage cell line. Transmission Electron Microscopy (TEM) and X-ray diffraction (XRD) used for the structural analysis of the drug reveals the presence of crystalline As2O3 nanoparticles of cubic structure. Similarly, signatures of M1-macrophage polarization were observed by surface enhanced Raman scattering (SERS) in RAW 264.7 cells with concomitant over expression of M1 cell surface marker, CD80 and transcription factor, NF-κB, respectively. We also observed a significant increase in pro-inflammatory cytokines like iNOS, TNF-α, IL-6, and COX-2 expression with unaltered ROS and apoptosis in drug-treated cells. Enhanced expression of Toll-like receptors 3 and 7 were observed both in transcriptional and translational levels after the drug treatment. In sum, our findings for the first time indicated the presence of crystalline As2O3 cubic nanostructure in Ars. alb 30C which facilitates modulation of innate immunity by activating macrophage polarization.

Effectiveness of Eupatorium perfoliatum 30C in Prevention of Dengue Fever and Acute Febrile Illness during 2017 Dengue Outbreak in Urban Slums of Delhi: A Prospective, Open-Label, Community-Based, Parallel Cohort Study.

OBJECTIVE: This study was undertaken to assess the effectiveness of Eupatorium perfoliatum (EP) 30C on the incidence of dengue fever and acute febrile illness (AFI) during the 2017 dengue outbreak. METHODS: We conducted a prospective, open-label, community-based parallel cohort study involving apparently healthy individuals residing in 06 urban slums (JJ colony) of Delhi. The participants were enrolled in two cohorts - the medicine cohort (MC) and the control cohort (CC). Participants in MC were given weekly one dose of EP 30C for 10 weeks along with Information, Education and Communication (IEC) material regarding dengue. Participants in the CC were provided with the IEC material only. The primary outcome measure was the incidence of dengue fever as per case definitions notified in the national guidelines for clinical management of dengue fever by the Government of India during the 10 weeks follow-up period. The secondary outcome measures were the incidence of AFI and the hospitalization of confirmed dengue cases. RESULTS: The analysis included 40,769 participants residing in 06 slum clusters of Delhi out of which 28,321 participants were in MC and 12,448 participants were in CC. The incidence of laboratory-confirmed dengue in the MC was 2.57 per 10,000 person-weeks (95% confidence interval [CI], 2.02-3.22) in comparison with 7.55 per 10,000 person-weeks (95% CI, 6.12-9.21) in the CC. The incidence of AFI in the MC was 19.66 per 10,000 person-weeks (95% CI, 18.07-21.36) in comparison with 40.96 per 10,000 person-weeks (95% CI, 37.48-44.67) in the CC. The overall protective effect of EP against laboratory-confirmed dengue was 65.77% (95% CI, 53.37-74.87; p = 0.0001) and against AFI was 52.58% (95% CI, 46.37-58.07; p = 0.0001). Hospitalization reported in the MC was nil as against 4.35% in the CC. No dengue-related case fatalities were reported from either cohort. None of the participants from the MC reported any adverse events owing to the prophylactic intervention. CONCLUSION: The study concludes that EP 30C was able to prevent dengue significantly. Randomized controlled trials are needed to confirm or refute our findings.Das Ziel dieser Studie war die Beurteilung der Wirksamkeit von Eupatorium perfoliatum (EP) 30C auf die Inzidenz von Dengue-Fieber und akuter fiebriger Erkrankung (AFE) während des Dengue-Ausbruchs 2017.Wir führten eine prospektive, unverblindete, Bevölkerungs-Parallelgruppen-Kohortenstudie mit augenscheinlich gesunden Bewohnern von 6 städtischen Slums (JJ-Kolonie) in Delhi durch. Die Teilnehmer wurden in 2 Kohorten aufgenommen, einer Medizinkohorte (MK) und einer Kontrollkohorte (KK). Die Teilnehmer in der MK erhielten 10 Wochen lang wöchentlich eine Dosis EP 30C und dazu Aufklärungsmaterialien über Dengue. Die Teilnehmer in der KK erhielten nur die Aufklärungsmaterialien. Die primäre Zielgröße war die Dengue-Fieber-Inzidenz laut der in den nationalen Leitlinien für das klinische Management des Dengue-Fiebers von der indischen Regierung bekannt gegebenen Falldefinition in dem zehnwöchigen Beobachtungszeitraum. Die sekundären Zielgrößen waren die Inzidenz von AFE und die Anzahl hospitalisierter bestätigter Dengue-Fälle.In die Analyse wurden 40,769 Bewohner von 6 Slum-Clustern in Delhi einbezogen, davon wurden 28,321 Teilnehmer in die MK aufgenommen und 12,448 Teilnehmer in die KK. Die Inzidenz von im Labor bestätigter Dengue betrug in der MK 2,57 pro 10,000 Personen/Woche (95%-Konfidenzintervall [KI]: 2,02­3,22), verglichen mit 7,55 pro 10,000 Personen/Woche (95%-KI: 6,12­9,21) in der KK. Die Inzidenz von AFI betrug in der MK 19,66 pro 10,000 Personen/Woche (95%-Konfidenzintervall [KI]: 18,07­21,36), verglichen mit 40,96 pro 10,000 Personen/Woche (95%-KI: 37,48­44,67) in der KK. Der Schutzeffekt (SE) von EP betrug gegen im Labor bestätigte Dengue 65,77% (95%-KI: 53,37­74,87; p = 0,0001) und gegen AFI 52,58% (95%-KI: 46,37­58,07; p = 0,0001). Die Hospitalisierungsrate war in der MK gleich Null versus 4,35% in der KK. In keiner Kohorte waren Dengue-bedingte Todesfälle zu verzeichnen. Bei keinem der Teilnehmer in der MK traten jegliche unerwünschten Ereignisse infolge der prophylaktischen Maßnahme auf.Die Studie gelangt zu dem Schluss, dass Eupatorium perfoliatum 30C in signifikantem Maße Dengue vorbeugen konnte. Randomisierte kontrollierte Studien sind erforderlich, um unsere Ergebnisse zu bestätigen bzw. zu widerlegen.

Tumor-associated macrophages: an effective player of the tumor microenvironment.

Cancer progression is primarily caused by interactions between transformed cells and the components of the tumor microenvironment (TME). TAMs (tumor-associated macrophages) make up the majority of the invading immune components, which are further categorized as anti-tumor M1 and pro-tumor M2 subtypes. While M1 is known to have anti-cancer properties, M2 is recognized to extend a protective role to the tumor. As a result, the tumor manipulates the TME in such a way that it induces macrophage infiltration and M1 to M2 switching bias to secure its survival. This M2-TAM bias in the TME promotes cancer cell proliferation, neoangiogenesis, lymphangiogenesis, epithelial-to-mesenchymal transition, matrix remodeling for metastatic support, and TME manipulation to an immunosuppressive state. TAMs additionally promote the emergence of cancer stem cells (CSCs), which are known for their ability to originate, metastasize, and relapse into tumors. CSCs also help M2-TAM by revealing immune escape and survival strategies during the initiation and relapse phases. This review describes the reasons for immunotherapy failure and, thereby, devises better strategies to impair the tumor-TAM crosstalk. This study will shed light on the understudied TAM-mediated tumor progression and address the much-needed holistic approach to anti-cancer therapy, which encompasses targeting cancer cells, CSCs, and TAMs all at the same time.

Prevalence and correlates of multimorbidity among patients attending AYUSH primary care settings in Delhi-National Capital Region, India.

INTRODUCTION: India has a multifaceted healthcare system and recognizes complementary and alternative systems of medicine (AYUSH) that cater to the healthcare needs of people. Multimorbidity requires frequent visits to physicians and long-term use of medications, due to which people tend to prefer AYUSH systems as they provide holistic patient-centered treatment. Hence, we aimed to estimate the prevalence of multimorbidity and assess its correlates among patients attending AYUSH primary care clinics in Delhi. METHODS: A cross-sectional study was conducted among 943 patients aged ≥ 18 years attending various AYUSH primary care clinics in Delhi from September 2021 to February 2022, employing a stratified random sampling technique. Descriptive statistics such as frequency and proportion were used to report the prevalence of multimorbidity (two or more chronic conditions in an individual out of the 33 conditions listed as per the Multimorbidity Assessment Questionnaire for Primary Care). A multivariable logistic regression assessed the association between various socio-demographic characteristics and multimorbidity, presented as an adjusted odds ratio (AOR) with a 95% confidence interval (CI). RESULTS: The prevalence of diabetes (14.7%) was found to be the highest (out of all included chronic conditions) among the patients attending various AYUSH primary care settings. The overall prevalence of multimorbidity was observed to be around 39.4%. We observed a higher likelihood of having multimorbidity among participants aged ≥ 70 years [AOR: 9.19 (95% CI: 3.75-22.54)], females [AOR: 1.57 (95% CI: 1.04-2.37)], and middle class [AOR: 2.23 (95% CI: 1.45-3.43)]. CONCLUSION: Multimorbidity was evidently prevalent across AYUSH primary care settings, which cannot be overlooked. The results suggest behavioral change communication may be aimed at older individuals, females, and the middle class.

Ultradiluted SARS-CoV-2 Spike Protein mitigates hyperinflammation in lung via ferritin and MMP-9 regulation in BALB/c mice.

AIM: This study investigated the prophylactic and therapeutic role of ultradiluted preparation of the Delta variant of SARS-CoV-2 recombinant spike (S) protein during S antigen-induced inflammatory process of disease progression along with the probable mechanism of action. MAIN METHODS: Ultradiluted S protein (UDSP) was prepared and administered orally to adult BALB/c mice before and after administration of S antigen intranasally. After an observation period of 72 h, animals were sacrificed and expression level of ferritin was assayed through ELISA. The genetic expressions of cytokines, IL-6, IL-10, IL-1ß, TNFα, IL-17, MMP-9, TIMP-1, ferritin light and heavy chains, and mitochondrial ferritin from lung tissues were investigated through RT-PCR. Formalin-fixed lung tissue sections were stained with hematoxylin and eosin to observe the degree of pathological changes. The activity of MMP-9 in lung tissues was investigated through gelatin zymography and immunofluorescence of MMP-9 in lung tissue sections was performed to revalidate the finding from gelatin zymography. Systems biology approach was used to elucidate a probable pathway where UDSP attenuated the inflammation through the regulation of pro- and anti-inflammatory cytokines. KEY FINDINGS: UDSP attenuated the S antigen-induced hyperinflammation in the lung by regulating pro- and anti-inflammatory cytokines, calming cytokine storm, reducing ferritin level both in transcriptional and translational levels, and restoring critical ratio of MMP-9: TIMP-1. SIGNIFICANCE: Our findings suggest a probable pathway by which UDSP might have attenuated inflammation through the regulation of cytokines, receptors, and other molecules. This proclaims UDSP as a promising antiviral agent in the treatment of COVID-19-induced immunopathogenesis.

Antiviral and anti-inflammatory activity of novel belladonna formulation against Japanese encephalitis virus via inhibition of p65 nuclear translocation and TNF-α mediated NF-kB signaling.

Japanese encephalitis (JE) is a mosquito-borne flavivirus infection, a major cause of viral encephalitis in South-East Asia with a CFR of ~30% and no specific treatment. Therefore, a novel belladonna formulation (BCT) was prepared and its antiviral and anti-inflammatory activity was elucidated during Japanese encephalitis virus (JEV) infection. Anti-JEV role of BCT was investigated aiming to prevent the infection in the peripheral immune cells. Antiviral activity of BCT was evaluated by plaque reduction assay, cell survival and apoptosis assay. BCT-mediated reduction in JEV-envelope expression was measured by indirect immunofluorescence, RT-PCR and Western blot assays. NF-κB expression and p65 nuclear translocation assays were determined to explore the mechanism of the action of BCT. TNF-α level was measured to evaluate the anti-inflammatory role of BCT during JEV infection. Consequently, molecular docking was performed with the TRAF2-TRADD complex. Our data suggested that BCT treatment reduces the JEV-plaque formation, JEV-induced cytopathic effects and increases cell survival. The antiviral effect of BCT was confirmed by reduction in the JEV-envelope protein expression. Moreover, BCT treatment and prevents the NF-κB activation via preventing the nuclear translocation of p65 and reduces the TNF-α levels. Our molecular docking analysis suggested that belladonna alkaloids interfere with the TRAF2-TRADD complex that results in inhibition of TNF-induced NF-κB signaling. For the first time, our data suggested that BCT reduces JEV expression and interferes with TNF-induced NF-κB signaling, thereby increasing cell survival via preventing the p65 nuclear translocation and may be used for the treatment and prevention of JE.Abbreviation: CFR: Case fatality rate; CAM: Complementary and alternative medicines; COX-2: Cyclooxygenase-2; IκB: Inhibitor kappa B; JE: Japanese encephalitis; JEV: Japanese encephalitis virus; NF-κB: Nuclear factor kappa-light-chain-enhancer of activated B cells; ORF: Open reading frame; TNFR: Tumor necrosis factor receptor; TNF-α: Tumor necrosis factor-α; TRADD: TNFR1-associated death domain protein; TRAF2: TNF Receptor Associated Factor 2.

Impact of COVID-19 Pandemic on Patients with Non-Communicable Disease: An Observational Cross-Sectional Study at AYUSH Set-Ups of Krishna and Darjeeling District, India.

OBJECTIVE: The objective of this study was to determine the impact of COVID-19 in patients suffering from NCDs in terms of their knowledge, awareness, perception about COVID-19, use of AYUSH immune boosters (AIB), and management of chronic condition during the pandemic. METHOD: During the unlock down period (October 2020), a cross-sectional study was conducted in the Krishna and Darjeeling district of Andhra Pradesh and West Bengal, India. 499 individuals suffering from at least one chronic disease were interviewed using a structured questionnaire. Logistic regression was applied to investigate the relationship of socio-demographic characteristics, AIB, and morbidity with pandemic-related care challenges. Principal component analysis was applied to minimize the dimensionality of factors related to COVID care challenges. RESULTS: 499 individuals were surveyed. 91% identified at least three correct COVID appropriate behaviours. 92.2% considered the coronavirus to be a potential threat (mean ± SD: 5.8 ± 2.6). 44.7% and 55.3% lived with one and 2 or more chronic conditions, respectively. Hypertension alone (27.4%) and diabetes with hypertension (33%) were leading presentations. Out of 499, participants, 88.8% had at least one form of AIB. 52% took Ars. alb. with other AIB and 40% took Ars. alb. alone. Only 9 participants were infected with COVID-19. CONCLUSION: In the interest of a densely populated country like India, the inclusion of simple and safe AYUSH measures is realistic, ethical, and cost-effective. AYUSH interventions as COVID-19 prophylactic and treatment as well as integrative care of chronic illnesses such as NCDs are suggested.

Efficacy of Arsenicum album 30C in the Prevention of COVID-19 in Individuals Residing in Containment Areas: A Prospective, Multicenter, Cluster-Randomized, Parallel-Arm, Community-Based, Open-Label Study.

OBJECTIVE: During the early part of the COVID-19 pandemic, non-pharmacologic interventions were the strategies for the prevention of severe acute respiratory syndrome coronavirus 2. The Ministry of Ayush, Govt. of India, had advised Arsenicum album 30C as a prophylactic to prevent COVID-19. This study was undertaken to evaluate the protective efficacy and safety of the Arsenicum album 30C. METHODS: We conducted a prospective, multicenter, cluster-randomized, parallel-arm, community-based, open-label study involving apparently healthy individuals residing in containment areas of 7 cities in India. Clusters are defined as the population residing in the containment areas, who are under restriction for movement. Forty-two clusters were randomly assigned at 2:1 to the Arsenicum album 30C group (30 clusters) or to the control group (12 clusters, which received no specific therapy). The medicine was given twice daily for 7 days. The primary outcome was the incidence of COVID-19, as per the case definition notified by the National Centre for Disease Control, Government of India, during 3-week follow-up period. RESULTS: The analysis included 32,186 individuals residing in 42 clusters (containment areas). A total of 22,693 individuals from 30 clusters received Arsenicum album 30C, and 9,493 individuals from 12 clusters were observed in the control group. The overall protective effect of the Arsenicum album 30C was 80.22% (95% confidence interval [CI], 71.16-86.44; 40 cases per 22,693 [6.04 per 10,000 person-weeks] in the Arsenicum album 30C group vs. 84 cases per 9,493 [29.78 per 10,000 person-weeks] in the control group). The protective effect of the Arsenicum album 30C against laboratory-confirmed COVID-19 was 68.22% (95% [CI], 49.64-80; 32 cases per 22,693 [4.83 per 10,000 person-weeks] in the Arsenicum album 30C group vs. 42 cases per 9,493 [14.93 per 10,000 person-weeks] in the control group). Adverse effects observed in both groups were mild and resolved without medication and sequelae. CONCLUSION: Homeopathic medicine Arsenicum album 30C was associated with a decrease in the incidence and provided some protection against COVID-19 as compared to nontreatment. Further randomized, double-blind, placebo-controlled trials may be conducted to validate the results of this study.In der Anfangszeit der COVID-19-Pandemie setzte man auf nicht-medikamentöse Massnahmen zur Prävention von Infektionen mit SARS-CoV-2 (Schweres-Akutes-Respiratorisches-Syndrom-Coronavirus 2). Das Ayush-Ministerium der indischen Regierung empfahl Arsenicum album 30C als Prophylaxe zur COVID-19-Prävention. Das Ziel dieser Studie war die Beurteilung der Schutzwirkung und Sicherheit von Arsenicum album 30C.An unserer prospektiven, multizentrischen, Cluster-randomisierten, unverblindeten Parallelgruppen-Bevölkerungs-Studie nahmen gesund erscheinende Bewohner von Sperrzonen in sieben indischen Städten teil. Die Cluster waren definiert als die Bewohnerpopulation je einer Sperrzone, für welche Mobilitätsbeschränkungen galten. 42 Cluster wurden per Randomisierung im Verhältnis 2:1 dem Arsenicum-album-30C-Arm (30 Cluster) oder dem Kontrollarm (12 Cluster, die keine spezifische Behandlung erhielten) zugeteilt. Das Medikament wurde sieben Tage lang zweimal täglich angewendet. Die primäre Zielgröße war die COVID-19-Inzidenz laut der vom National Centre for Disease Control der indischen Regierung bekannt gegebenen Falldefinition in einem Nachbeobachtungszeitraum von drei Wochen.Die Analyse umfasste 32.186 Personen in 42 Clustern (Sperrzonen). Insgesamt 22.693 Personen in 30 Clustern erhielten Arsenicum album 30C, und 9.493 Personen in 12 Clustern wurden als Kontrollarm beobachtet. Die Ergebnisse für Alter, Geschlecht und Komorbidität waren im Behandlungs- und Kontrollarm vergleichbar. Die Gesamt-Schutzwirkung von Arsenicum album 30C betrug 80,22 % (95-%-Konfidenzintervall [KI]: 71,16­86,44; 40 Fälle unter 22.693 [6,04 pro 10.000 Personen-Wochen] im Arsenicum-album-30C-Arm vs. 84 Fälle unter 9.493 [29,78 pro 10.000 Personen-Wochen] im Kontrollarm). Die Schutzwirkung von Arsenicum album 30C gegen laboranalytisch bestätigte COVID-19-Erkrankung betrug 68,22 % (95-%-KI: 49,64­80; 32 Fälle unter 22.693 [4,83 pro 10.000 Personen-Wochen] im Arsenicum-album-30C-Arm vs. 42 Fälle unter 9.493 [14,93 pro 10.000 Personen-Wochen] im Kontrollarm). Die beobachteten unerwünschten Wirkungen waren in beiden Gruppen von geringer Schwere und klangen ohne Medikation folgenlos ab.Das Homöopathikum Arsenicum album 30C war im Vergleich zur Nichtbehandlung mit einer Verringerung der Inzidenz assoziiert und bot gewissen Schutz vor COVID-19. Weitere randomisierte, doppelblinde, placebokontrollierte Studien könnten durchgeführt werden, um die Ergebnisse dieser Studie zu validieren.

Homoeopathic Medicine Arsenicum album 30 C for Covid-19 in 2020: A Retrospective Analysis from Mass-level Data.

Background and Objective: Homoeopathy has played a notable role in managing epidemics in the past. The Ministry of Ayush, Government of India, declared Arsenicum album 30 C as a prophylactic for Covid-19, which was followed by the distribution of the medicine across India. The Central Council for Research in Homoeopathy (CCRH) collected post-prophylactic consumption data of individuals from various colleges over months, which created a data pool. Considering the importance of these mass-level data and their possible impact on public healthcare decisions, the information gathered from this heterogeneous population cohort was subjected to a retrospective data analysis to observe the incidence of Covid-19 in the community. Methods: Data from 50 colleges from February-August 2020 showed that 10.6 million people in 13 states of India received prophylactic medicine during the study period. The data was collected from individuals three weeks following prophylactic consumption for a retrospective analysis. The incidence of Covid-19 was assessed. Results: The data of 584 980 individuals who met the study criteria were included in the analysis. The incidence of Covid-19 in the population cohort was 13.58 per 10 000-person weeks (95% CI, 13.04 to 14.14), which remained near-constant over time despite the increasing disease burden in the country (12.87 to 14.52 per 10 000-person weeks). Consumption of the prophylactic significantly reduced the risk of contracting Covid-19 in high-risk groups as compared to their counterparts. Conclusion: The study concludes that Arsenicum album 30 C has a potential prophylactic effect against Covid-19. Further controlled studies are recommended to establish a causal relation.

Effectiveness of Arsenicum album 30C in Prevention of COVID-19 in Individuals Residing in Containment Zones of Delhi-A Prospective, Community-based, Parallel Cohort Study.

OBJECTIVE: This work was undertaken to evaluate the protective effect of Arsenicum album 30C against COVID-19. DESIGN: The work was designed as a prospective parallel cluster cohort study. INTERVENTION: Participants were enrolled in a homeopathy intervention (HI) cohort (who received Arsenicum album) or in a non-intervention (NI) cohort (who received no systematic intervention) from COVID-19 containment areas of Delhi. Individuals of age 5 years or above were given four medicated pills of Arsenicum album 30C, while those from 1 to 5 years old were given two medicated pills in each dose. RESULTS: The analysis included 10,180 individuals residing in 11 COVID-19 containment areas in Delhi, out of which 6,590 individuals were in the HI cohort and 3,590 individuals were in the NI cohort. The overall protective effect of Arsenicum album 30C was 83.43% (95% confidence interval [CI], 76.77 to 88.17): 45 cases per 6,590 (8.34 per 10,000 person-weeks) in the Arsenicum album 30C group versus 143 cases per 3,590 (45.01 per 10,000 person-weeks) in the NI cohort. The protective effect of Arsenicum album 30C against laboratory confirmed COVID-19 was 74.40% (95% CI, 55.08 to 85.41): 18 cases per 6,590 (3.32 per 10,000 person-weeks) in the Arsenicum album 30C group versus 38 cases per 3,590 (11.85 per 10,000 person-weeks) in the NI cohort. CONCLUSION: The use of Arsenicum album 30C was associated with some protection against probable and laboratory-confirmed COVID-19 in a containment-zone setting. Randomized controlled trials are needed to confirm or refute these results.

Preparation, Standardization and Anti-plasmodial Efficacy of Novel Malaria Nosodes.

BACKGROUND: Resistance to artemisinin and its partner drugs has threatened the sustainability of continuing the global efforts to curb malaria, which urges the need to look for newer therapies to control the disease without any adverse side effects. In the present study, novel homeopathic nosodes were prepared from Plasmodium falciparum and also assessed for their in vitro and in vivo anti-plasmodial activity. METHODS: Three nosodes were prepared from P. falciparum (chloroquine [CQ]-sensitive [3D7] and CQ-resistant [RKL-9] strains) as per the Homeopathic Pharmacopoeia of India, viz. cell-free parasite nosode, infected RBCs nosode, mixture nosode. In vitro anti-malarial activity was assessed by schizont maturation inhibition assay. The in vitro cytotoxicity was evaluated by MTT assay. Knight and Peter's method was used to determine in vivo suppressive activity. Mice were inoculated with P. berghei-infected erythrocytes on day 1 and treatment was initiated on the same day. Biochemical, cytokine and histopathological analyses were carried out using standard methods. RESULTS: In vitro: the nosodes exhibited considerable activity against P. falciparum with maximum 71.42% (3D7) and 68.57% (RKL-9) inhibition by mixture nosode followed by cell-free parasite nosode (62.85% 3D7 and 60% RKL-9) and infected RBCs nosode (60.61% 3D7 and 57.14% RKL-9). The nosodes were non-toxic to RAW macrophage cell line with >70% cell viability. In vivo: Considerable suppressive efficacy was observed in mixture nosode-treated mice, with 0.005 ± 0.001% parasitemia on day 35. Levels of liver and kidney function biomarkers were within the normal range in the mixture nosode-treated groups. Cytokine analysis revealed increased levels of IL-4 and IL-10, whilst a decline in IL-17 and IFN-γ was evident in the mixture nosode-treated mice. CONCLUSION: The mixture nosode exhibited promising anti-malarial activity against P. falciparum and P. berghei. Biochemical and histopathological studies also highlighted the safety of the nosode for the rodent host. The study provides valuable insight into a novel medicament that has potential for use in the treatment of malaria.

Efficacy of Chininum Sulphuricum 30C against Malaria: An in vitro and in vivo Study.

BACKGROUND: New effective, economical and safe antimalarial drugs are urgently needed due to the development of multi-drug-resistant strains of the parasite. Homeopathy uses ultra-diluted doses of various substances to stimulate autoregulatory and self-healing processes to cure various ailments. The aim of the study was to evaluate the in vitro and in vivo antimalarial efficacy of a homeopathic drug, Chininum sulphuricum 30C. METHODS: In vitro antiplasmodial activity was screened against the P. falciparum chloroquine-sensitive (3D7) strain, and cell viability was assessed against normal human dermal fibroblasts and HepG2 cells. Suppressive, preventive and curative studies were carried out against P. berghei-infected mice in vivo. RESULTS: Chininum sulphuricum (30C) revealed good antiplasmodial activity in vitro, with 92.79 ± 6.93% inhibition against the 3D7 strain. The cell viability was 83.6 ± 0.6% against normal human dermal fibroblasts and 95.22 ± 5.1% against HepG2 cells. It also exhibited suppressive efficacy with 95.56% chemosuppression on day 7 with no mortality throughout the follow-up period of 28 days. It also showed preventive activity against the disease. Drug treatment was also safe to the liver and kidney function of the host as evidenced by biochemical studies. CONCLUSION: Chininum sulphuricum 30C exhibited considerable antimalarial activity along with safety to the liver and kidney function of the host.

In-vitro antiviral action of Eupatorium perfoliatum against dengue virus infection: Modulation of mTOR signaling and autophagy.

ETHNOPHARMACOLOGICAL RELEVANCE: Dengue virus (DENV) is a re-emerging mosquito-borne flavivirus that has recently engendered large epidemics around the world. Consequently antivirals with effective anti-DENV therapeutic activity are urgently required. In the 18th century, Europeans, as well as native inhabitants of North America, were known to adapt the medicinal property of the common perennial plant Eupatorium perfoliatum L. to treat fever and infections. Previous studies have shown that Eupatorium perfoliatum L. possesses anti-inflammatory, anti-oxidative, anti-plasmodial, anti-bacterial and antiviral activities. However, to the best of our knowledge, no anti-DENV activity of E. perfoliatum L. has been investigated at the molecular level so far. AIM OF STUDY: Here, for the first time we have attempted to study the action of E. perfoliatum extract and its few bioactive components i.e., quercetin, caffeic acid and eupafolin against wild primary clinical isolate of DENV-2 infection in an in vitro model. MATERIALS AND METHODS: The presence of the bioactive components in the E. perfoliatum extract, were analyzed by HPLC- DAD. Then, CC50 as well as IC50 values of the extract and its bioactive components were measured against DENV in HepG2 cell line. After that, the antiviral activity was studied by Time of addition assay using qRT-PCR. Further, the downstream signalling action of E. perfoliatum extract, was studied by Human phosphorylation MAPK antibody array, followed by immunofluorescence microscopy. Moreover, a molecular docking analysis was done to study the binding affinity of bioactive components of E. perfoliatum extract with TIM-1 transmembrane receptor protein, which is known for viral internalization. RESULT: We found that E. perfoliatum extract has marked antiviral activity during pre-treatment against DENV infection in HepG2 cell line. The extract also significantly reduced the DENV induced autophagy in HepG2 cell line as detected by LC3 II localization. The presence of different bioactive compounds in E. perfoliatum extract were confirmed by HPLC-DAD. In the bioactive components, in parallel to earlier studies, quercetin showed the most significant preventive action against DENV infection. Further, in molecular docking analysis also, quercetin showed the strongest binding affinity towards DENV membrane receptor TIM-1 protein. CONCLUSION: Our findings suggests that E. perfoliatum extract has significant potential to be an anti-DENV therapeutic agent. Moreover, among the bioactive components, quercetin may have a prophylaxis role in executing the antiviral activity of E. perfoliatum extract against DENV infection.

Evidence for Reversal of Immunosuppression by Homeopathic Medicine to a Predominant Th1-type Immune Response in BALB/c Mice Infected with Leishmania donovani.

BACKGROUND: Visceral leishmaniasis (VL) is a neglected tropical disease that is fatal if treatment is not given. The available chemotherapeutic options are unsatisfactory, and so complementary therapies like homeopathy might be a promising approach. METHODS: A nosode from a pure axenic culture of Leishmania donovani was prepared and screened for its anti-leishmanial potential both in an in-vitro and an in-vivo experimental approach. RESULTS: Leishmania donovani amastigote promastigote nosode (LdAPN 30C) exhibited significant anti-leishmanial activity against the promastigote forms of Leishmania donovani and was found to be safe. A study conducted on VL-infected mice revealed that LdAPN 30C resolved the disease by modulating the host immune response toward the Th1 type through upregulating the pro-inflammatory cytokines (IFN-γ and IL-17) and inducing nitric oxide (NO) levels in the infected macrophages. The hepatic parasite load was also found to be significantly decreased. The nosode was found to be safe, as no histological alterations in the liver or kidney were observed in the animals treated with the LdAPN 30C. CONCLUSION: This is the first study in which an axenic culture of Leishmania donovani has been used for the preparation of a homeopathic medication. The study highlights the anti-leishmanial and immunomodulatory potential of a homeopathic nosode in experimental VL.

Stem cell therapy: A paradigm shift in breast cancer treatment.

As per the latest Globocan statistics, the high prevalence rate of breast cancer in low- and middle-income countries has led to it becoming the most common cancer to be diagnosed, hence posing a major public health challenge. As per this data, more than 11.7% of the estimated new cancer cases in 2020 were due to breast cancer. A small but significant subpopulation of cells with self- renewing ability are present in the tumor stroma and have been given the nomenclature of cancer stem cells (CSCs). These cells display a high degree of plasticity owing to their ability to transition from the slowly cycling quiescent phase to the actively proliferating phenotype. This attribute of CSCs allows them to differentiate into various cell types having diverse functions. Breast CSCs have a pivotal role in development, metastasis, treatment resistance and relapse of breast cancers. This review focuses on the pathways regulating breast CSC maintenance and the current strategies that are being explored for directing the development of novel, targeted, therapeutic approaches for limiting and eradicating this aberrant stem cell population.

Clinical Repurposing of Medicines is Intrinsic to Homeopathy: Research Initiatives on COVID-19 in India.

As at mid-October 2020, the coronavirus disease 2019 (COVID-19) pandemic has been continuing on the rise across the globe, including in India. Historically, homeopathy has been used in a number of epidemics/pandemics. The development of homeopathic medicines is approached uniquely through "drug provings" and clinical verification; these two intrinsic processes establish the background for the application of homeopathic medicines, regardless of nosological diagnosis. This article reflects research initiatives on COVID-19 in India and identifies studies listed in the Clinical Trial Registry-India database. We identified 29 studies being undertaken in different settings, including those in conventional medicine: 20 randomized controlled trials (RCTs) and 9 observational studies. Fifteen studies are aimed at prophylaxis and 14 are aimed at treatment. Amongst the treatment studies, 11 are focused on efficacy or comparative effectiveness. The findings might provide evidence for clinically repurposing some of homeopathy's medicines, an approach that is intrinsic to the therapy, enabling their use in COVID-19 as an adjuvant or stand-alone to help reduce costs and improve patient recovery.