Ayurvedic Management of Hypothyroidism - A Case Report

Hypothyroidism is the significant endocrine problem found in general public. It is characterized by insufficient production of thyroid hormones, leading to abnormal levels. This condition prompts the decrease in Basal metabolic rate of the body. Ayurveda places a significant part to preclude the basic reason and to treat it. In Ayurveda, it tends to be associated with Kapha-vata dosha dushti, Rasavaha Strotasadushti, Medadushti and Agnimandya. In present study, a 22 years old female patient suffering from Sthaulya (~Weight gain), Duarbalya (~Weakness), Twak rukshata (~Dry skin), Kesh patana (hair loss), puffiness around eyes, mood swings, impaired memory, emotional depression, drowsiness, headache, Amalapitta (~Hyperacidity) and lethargy from last three years. She had been taking allopathic medicine without experiencing any satisfactory relief in her symptoms. In the present study, group of Herbo-mineral preparation was prescribed to the patient for a period of 4 months. After four months patient experienced relief, and her allopathic medicine was completely stopped. Keywords: Hypothyroidism, Agnimandya, Sthauly

Clinical Study and Assessment of Efficacy of Polyherbal Combination (KNDBHU) in COVID 19 Patients

Even three years after the COVID-19 pandemic initially appeared in December of the year 2019, it is still raging over the globe. There have been numerous attempts to use well-known medications in new ways to treat COVID-19 infection. Many nations, including India, implemented a series of lock downs in an effort to limit the pandemic. Sanitation practices, social seclusion, routine mask wearing, testing, and quarantining of suspected carriers were among the recommended methods for limiting the illness. Numerous treatment strategies have also been used to prevent and treat the illness, but no formal studies using well-known Ayurvedic formulations or any polyherbal combinations have been conducted. A clinical study was planned to test the noble polyherbal combination containing Withania somnifera, Tinospora cordifolia, Moringa oleifera, Adhatoda vasica, Piper longum, Glycyrrhiza glabra, Ocimum sanctum and Curcuma longa. The study checked the rate of symptom remission in individuals receiving the polyherbal combination in addition to standard of care (SoC) to that in patients with mild and moderate symptoms of COVID-19 infections receiving SoC alone. A prospective randomized interventional clinical study was planned comparing outcomes in 2 cohorts with mild to moderate COVID-19 as under. Cohort-1 was trialled with conventional treatment as per government advisory. Cohort-2 was trialled with polyherbal combination and with conventional treatment as per government advisory. With the current add-on Polyherbal regimen, an early clinical improvement in breathlessness was seen along with early ageusia and cough reduction when compared with conventional treatment. The polyherbal combination (KNDBHU) enhanced recovery in COVID-19 Patients. Given the lack of hospital beds in India, the median length of hospital stays was shortened; this development is significant. Keywords: Covid­19, Add on polyherbal combinationregimen for Corona virus, Early Clinical Improvement, Shortened hospital stay

Evaluation of Rasayaṇa activity of Rudanti (Capparis moonii Wight.) in the Management of Rajayakshma (Pulmonary tuberculosis)

It is estimated that about 1/3rd world’s population is infected by Mycobacterium tuberculosis. It kills more adults in India than any other infectious diseases. In 2018, India was able to achieve a Total Notification of 21.5 Lakh TB cases of which 25 % was from the private sector. Majority of the TB burden is among the working age group. The 89% of TB cases come from the age group of 15-69 years. About 2/3 of the TB cases are Males. Uttar Pradesh, with 17% of population of the country, is the largest contributor to the TB cases in with 20% of the total notifications, accounting to about 4.2 Lakh cases (187 cases/ lakh population). Aim and objective: This study was intended to evaluate the rasayaṇa effect of Rudanti (Capparis moonii wight.) as an adjunct to anti-tubercular drugs in the management of Rajayakshma (PTB). Methodology: This is a single blind therapeutic control randomized clinical trial consisting of 100 patients divided in to two groups. Group A patients were treated with DOTS and group B were treated with DOTS along with Rudanti powder 5 gm BD for 6 months. Conclusion: This study shows that by incorporating an herbal rejuvenative in the management of PTB, a chronic debilitating disorder, had an additive effect. Rudanti powder can be easily administered as an adjunct to DOTS. Rudanti has hepatoprotective properties, which remunerate the hepatotoxicity known to cause by DOTS drugs. Improved weight gain in TG patients in comparison to CG indicates the accessory effect of Rudanti

The scientific basis of rasa (taste) of a substance as a tool to explore its pharmacological behavior

Background: A rational and well-developed pharmacological basis forms the foundation of therapeutics in Ayurveda. The principles and theories of Ayurveda need to be validated in the scientific context in order to harness the millennia old knowledge. Rasa (taste) of the substance is the foremost tool in Ayurveda to assess and determine the pharmacological properties and actions of the substance. Similarity in rasa is said to signify similar structure and consequently similar pharmacological behavior. Depending on skills developed over the course of long-term clinical experience one, can register the minute variations in rasa of substances and accordingly the possible variations in pharmacological actions. Thus, rasa can be used as a scientific tool in the drug discovery process to limit and focus the target areas. Aim: To sensitize scientific community to the utility of rasa as a tool in the process of drug discovery. Materials and Methods: All relevant ancient and contemporary literature was reviewed critically to form a scientific basis of the Ayurvedic concept of rasa as a tool to identify the pharmacological behavior of a substance. Conclusion: The review finds that rasa (taste) can be used as a guide to identify potential targets in drug discovery

HSP70 Domain II of <i>Mycobacterium tuberculosis</i> Modulates Immune Response and Protective Potential of F1 and LcrV Antigens of <i>Yersinia pestis</i> in a Mouse Model

<div><p>No ideal vaccine exists to control plague, a deadly dangerous disease caused by <i>Yersinia pestis</i>. In this context, we cloned, expressed and purified recombinant F1, LcrV antigens of <i>Y. pestis</i> and heat shock protein70 (HSP70) domain II of <i>M. tuberculosis</i> in <i>E. coli</i>. To evaluate the protective potential of each purified protein alone or in combination, Balb/C mice were immunized. Humoral and cell mediated immune responses were evaluated. Immunized animals were challenged with 100 LD₅₀ of <i>Y. pestis</i> via intra-peritoneal route. Vaccine candidates <i>i.e.</i>, F1 and LcrV generated highly significant titres of anti-F1 and anti-LcrV IgG antibodies. A significant difference was noticed in the expression level of IL-2, IFN-γ and TNF-α in splenocytes of immunized animals. Significantly increased percentages of CD4+ and CD8+ T cells producing IFN-γ in spleen of vaccinated animals were observed in comparison to control group by flow cytometric analysis. We investigated whether the F1, LcrV and HSP70(II) antigens alone or in combination can effectively protect immunized animals from any histopathological changes. Signs of histopathological lesions noticed in lung, liver, kidney and spleen of immunized animals on 3^rd day post challenge whereas no lesions in animals that survived to day 20 post-infection were observed. Immunohistochemistry showed bacteria in lung, liver, spleen and kidney on 3^rd day post-infection whereas no bacteria was observed on day 20 post-infection in surviving animals in LcrV, LcrV+HSP70(II), F1+LcrV, and F1+LcrV+HSP70(II) vaccinated groups. A significant difference was observed in the expression of IL-2, IFN-γ, TNF-α, and CD4+/CD8+ T cells secreting IFN-γ in the F1+LcrV+HSP70(II) vaccinated group in comparison to the F1+LcrV vaccinated group. Three combinations that included LcrV+HSP70(II), F1+LcrV or F1+LcrV+HSP70(II) provided 100% protection, whereas LcrV alone provided only 75% protection. These findings suggest that HSP70(II) of <i>M. tuberculosis</i> can be a potent immunomodulator for F1 and LcrV containing vaccine candidates against plague.</p></div

Histopathology of the organs collected from the immunized group animals on 3^rd and 20^th day post infection with <i>Y. pestis</i> and the naive control animals that were neither immunized nor challenged with <i>Y. pestis</i>.

<p>Tissue sections were stained with hematoxylin and eosin for pathological examination. Tissue section collected from naive control and immunized animals on 3^rd day post infection <i>i.e.</i>, Naive control (A); PBS control (B); HSP70(II) (C); F1 (D); F1+HSP70(II) (E); LcrV (F); LcrV+HSP70(II) (G); F1+LcrV (H); F1+LcrV+HSP70(II) (I). Tissue sections were collected from the survived animal groups on 20^th day post infection <i>i.e.</i>, LcrV (J); LcrV+HSP70(II) (K); F1+LcrV (L); F1+LcrV+HSP70(II) (M). Photomicrograph represents the histopathology of Lung[<b>a</b>]: the arrows in the panel B indicate the infiltration of neutrophils. Photomicrograph of spleen [<b>b</b>]: in the panel B, reduced density of white pulp follicle and congestion in the red pulp, lymphoid follicle depletion shown by arrow and the presence of megakaryocytes shown by bold arrow. Photomicrograph of kidney [<b>c</b>]: the granular degeneration of parenchyma was observed in the panel B (bold arrows) and swelling in renal tubules (arrows). Photomicrograph of liver [<b>d</b>]: in the panel B, the hepatocytes degeneration was observed as indicated by arrow.</p

Measurement of serum IgG antibody titers in immunized Balb/C mice.

<p>[A] Sera collected after first booster (14^th day) and second boosters (21^st day) from immunized groups (F1, F1+HSP70(II), F1+LcrV, F1+LcrV+HSP70(II)) were measured for F1-specific IgG by indirect ELISA. [B] Determination of LcrV-specific serum IgG antibody titer in the sera from immunized groups (LcrV, LcrV+HSP70(II), F1+LcrV, F1+LcrV+HSP70(II)). Data are represented in mean antibody titers with SD of eight Balb/C mice in each group. The cut-off value for the assays was calculated as the mean OD (+2 SD) from sera of control group assayed at 1∶100 dilution. Serum endpoint IgG titers were calculated as the reciprocal of the highest serum dilution giving an OD more than the cut-off. Analysis was done by one way ANOVA, All Pairwise Multiple Comparison Procedure (Fisher LSD Method). ** P<0.01; *** P<0.001; ^# P<0.001.</p

Immunohistochemistry (IHC) staining for localization of <i>Y. pestis</i> in the organs collected from immunized group animals on 3^rd and 20^th day post infection with <i>Y. pestis</i> and the naive control animals that were neither immunized nor challenged.

<p>The F1 antigen of <i>Y. pestis</i> was identified with anti-mouse FITC conjugated secondary antibody in the tissue sections collected from immunized animal groups on 3^rd day post infection including naive control <i>i.e.</i>, Naive control (A); PBS control (B); HSP70(II) (C); F1 (D); F1+HSP70(II) (E); LcrV (F); LcrV+HSP70(II) (G); F1+LcrV (H); F1+LcrV+HSP70(II) (I). Tissue sections were collected from the survived animal groups on 20^th day post infection <i>i.e.</i>, LcrV (J); LcrV+HSP70(II) (K); F1+LcrV (L); F1+LcrV+HSP70(II) (M). Fluorescent images representing the localization of <i>Y. pestis</i> in tissue sections of Lung [<b>a</b>]; Spleen [<b>b</b>]; Kidney [<b>c</b>]; and Liver [<b>d</b>].</p

a. Schematic diagram of three recombinant vaccine candidates; F1, LcrV and HSP70(II) showing the histidine tag and orientation of the open reading frame.

<p><b>b</b>. 16% SDS-PAGE analysis of F1 protein expression [<b>A</b>]. 12% SDS–PAGE analysis of LcrV [<b>B</b>] and of HSP70(II) domain II of <i>M. tuberculosis</i> protein expression in <i>E. coli</i> [<b>C</b>]. The panels depict protein molecular mass marker (lane M), and Coomassie-stained polypeptide profiles of <i>E. coli</i> lysates un-induced (lane U) and induced with IPTG (lane I). The arrows at the right of the panels indicate the position of expressed recombinant proteins. <b>c</b>. SDS-PAGE analysis of purified F1 [<b>A</b>], LcrV [<b>B</b>] and HSP70(II) domain II of <i>M. tuberculosis</i> [<b>C</b>] metal affinity chromatography using Ni-NTA column. Each purified protein (3 µg/well) was analysed on SDS-PAGE. <b>d</b>. The humoral and cell mediated immune responses, protective potential and histopathological examinations of F1 and LcrV from <i>Y. pestis</i> with or without HSP70(II) of <i>M. tuberculosis</i> were evaluated in a mouse model. [<b>A</b>] Balb/C mice (8/group) were immunized with plague vaccine candidates with HSP70(II) as an immunomodulator in formulation aluminium hydroxide gel. [<b>B</b>] Schematic representation of immunization schedule following challenge experiments.</p

List of oligos used for Cloning of <i>caf1, lcrV</i> and <i>hsp70(II)</i> genes in pET28a⁺ vector.

<p>Homologous nucleotide sequences of <i>caf1, lcrV</i> and <i>hsp70(II)</i> in capital case and the engineered sequences (ata) at the 5′ ends are shown in small case. The in-frame initiator codon in the forward primer is shown in bold and the compatible restriction sites are underlined.</p><p>List of oligos used for Cloning of <i>caf1, lcrV</i> and <i>hsp70(II)</i> genes in pET28a⁺ vector.</p