Prashanth Varkey’s research while affiliated with Jubilee Mission Medical College and Research Institute and other places

Dermal substitutes are crucial in soft-tissue reconstruction, minimising donor-site morbidity. However, conventional protocols often involve multi-staged procedures. This case series explores the effectiveness of integrating dermal substitutes with skin grafting in a one-stage process for soft-tissue coverage, supplemented by hyperbaric oxygen (HBO) therapy. The study, conducted in the plastic surgery and burns department from June 2022 to December 2022, included 10 patients with soft-tissue defects. All underwent single-stage dermal substitute and skin grafting, accompanied by HBO therapy for soft-tissue reconstruction. Among the 10 patients, seven had soft-tissue defects in the leg region, comprising three with chronic ulcers, three with post-traumatic leg defects and one with a chronic ulcer due to peripheral vascular disease. Two patients had post-burn contracture release and reconstruction, while one presented with a soft-tissue defect following sternal wound dehiscence post-irradiation. All wounds healed successfully without complications, and all patients received six sessions of HBO therapy. The incorporation of HBO therapy as an adjunct proved beneficial in achieving successful single-stage reconstruction of soft-tissue defects using dermal substitutes and skin grafting.

Background: Urinary tract infections (UTIs) are some of the most commonly encountered infections in clinical practice that affects 150 million people each year worldwide. The emergence of resistance, adverse effects of antimicrobial agents, and other related issues have prompted the establishment of a research framework to explore alternative methods for managing UTIs. Cardamom exhibits unique botanical characteristics and biochemical compositions, contributing to their diverse culinary and medicinal applications. Techniques like gas chromatography-mass spectrometry analysis have identified 1,8-cineole and a-terpineol as main bioactive components in both black and green cardamom. Present study addresses urgent, unmet analytical and clinical care needs by exploring adoption of alternative therapies, specifically nutraceuticals viz, 1, 8-cineole and α-terpineol, major bioactive from cardamon for controlling and treating UTIs. Methods: Different concentrations of 1,8-cineole, α-terpineol and 1,8-cineole/α-terpineol (1:1) were used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against multi-drug resistant (MDR) E. coli isolated from UTIs. Results: The mean MIC and MBC of 1,8-cineole, α-terpineol and 1,8-cineole/α-terpineol (1:1) for MDR E. coli strains were 7.73 µg/ml, 0.32 µg/ml and 0.32 µg/ml respectively. For non MDR E. coli strains the mean MIC and MBC of the 1, 8-cineole, α-terpineol and 1,8-cineole/α-terpineol (1:1) were 2.14 µg/ml, 0.2 µg/ml and the 0.2 µg/ml respectively. Conclusions: 1,8-cineole and α-terpineol exhibited good antimicrobial activity against urinary E. coli isolates tested, including MDR strains.

Aim: To investigate anti-inflammatory, cytotoxic effects and viability of Recovereez- 51% 1,8-Cineol and 1,8-Cineol (natural), and to determine safety for both of the compounds and its use in new product range of nutraceuticals. Methodology: Cell line was divided into two groups; one was treated with Recovereez(R) and another one with 1,8-Cineol(C). The ELISA assay method was used to measure the COX, LOX, MPO, NO, and nitrate levels. Cellular viability was performed by observing cells under the inverted phase contrast microscope, followed by MTT assay. Results: R&C showed a reduction in levels of inflammatory markers such as IL-6, IL-1β, TNF-α, LOX, MPO, COX, iNOS, and nitrate levels. The inhibition as observed in the levels of COX and LOX enzymes were 65%±0.1 and 59%±0.3µg/ml, and 72%±0.3 and 67%±0.4µg/ml for R&C, respectively. iNOS production showed inhibition by 70%±0.3 and 65%±0.8µg/ml and NO production were reduced by 193 and 209.5µg/ml by R&C, respectively. Again, there was a decrease in the activity of MPO by 0.35 and 0.39U/ml for R&C, respectively. The results were significant with p<0.001 compared to the Diclofenac sodium. Cell viability of 60% was observed upon treating L929 cells with R&C. Conclusion: R&C exerted an anti-inflammatory effect on RAW 264.7 cells without exerting cytotoxicity in a dose-dependent manner. Therefore, Recovereez could be used as an alternative treatment or to prevent inflammatory diseases. Moreover, R&C are expected to treat different types of cancer. The findings could potentially lead to the discovery of safe and effective bioactive compounds that can prevent or cure the occurrence of cancerous cells. Keywords: dietary supplements, nutraceutical, COX, anti-inflammation, cancer

Aim Oral candidiasis is a fungal infection of the oral cavity caused by Candida albicans. Currently used antifungal drugs fail to treat oropharyngeal candidiasis. Herbal remedies are found to be safe and effective to treat bleeding gums, halitosis, mouth ulcers, and reduce tooth decay. Cardamom is known to possess antimicrobial, antioxidant, and immunomodulatory activity. In an in vitro study, we aim to evaluate the antifungal potential of cardamom-based Denteez TM mouthwash against opportunistic fungal infections in the oral cavity. Materials and Methods Five Candida species were included in the study: namely C. albicans, C. tropicalis, C. krusei, C. parapsilosis, and C. glabrata. Additionally, 54 clinical isolates obtained from oral cavity of healthy individuals and patients suffering from oral candidiasis were randomly selected and included. Various antifungal agents tested included fluconazole, itraconazole, ketoconazole, miconazole, and amphotericin B. Minimum inhibitory concentration (MIC) and time-kill assay were performed as per standard guidelines. Results The MIC obtained for fluconazole against different Candida species was 1.25 µg/ml. The MIC of azole drugs was in the range of 0.312–0.62 µg/ml. The MIC of Denteez TM mouthwash was determined as 1.25%. The clinical isolates of Candida showed major resistance to fluconazole (75.9%) and absolute susceptibility to amphotericin B. Denteez TM was found effective in inhibiting 98.2% of the clinical isolates. Denteez TM mouthwash showed time-dependent inhibition of all Candida species. Conclusion The mouthwash has comparable efficacy to that of other routinely used antifungal agents. Denteez TM can be effectively used to deal with oral candidiasis and antifungal resistance.

The SARS-CoV-2 instigated “cytokine storm” elicited upon infection is known to majorly cause lung injury and even mortality in severe cases. Early clinical prognosis to alleviate the exaggerated release of inflammatory cytokines is thus looked upon. Considering the recent attention and advantages of saliva as a clinical specimen, i.e. ease and painlessness of collection, which does not require trained staff and could allow self-sampling, the present study attempts to explore saliva for detection of IL-6, TNF-α and IL-10 which constitute major inflammatory genes that are elevated in COVID-19 using RT-PCR. Blood specimens of the same patients were also parallelly assessed to compare and validate the inflammatory marker expression. A total of 64 COVID-19 subjects who met the inclusion criteria were enrolled in this pilot study. Paired samples of blood and saliva from each patient were collected as per standard sampling protocols. RNA from all specimens were extracted using Qiagen RNA Blood Mini Kit and subjected to RT-PCR. IL-6, TNF-α and IL-10 expression were assessed in Ct (cycle threshold) values. It was observed that all 64 (100%) patients expressed IL-6 gene and TNF-α gene, whereas only 7 (5.19%) patients expressed IL-10 in both blood and saliva samples. The mean Ct values of IL-6 gene expressed in blood and saliva were 26.68 ± 2.26 and 28.53 ± 3.11 respectively. Similarly, the mean Ct values of TNF-α gene expressed in blood and saliva were 27.98 ± 2.45 and 28.92 ± 3.70 respectively. The observed mean Ct values of IL-10 gene expressed in blood and saliva were 31.26 ± 3.96 and 30.11 ± 4.12 respectively. Accordingly, the results indicate that inflammatory genes IL-6, TNF-α and IL-10 were detectable in both patient saliva as well as in blood. Moreover, mean Ct values of IL-6, TNF-α and IL-10 in both samples were found to be comparable. This finding thus suggests the possible use of saliva as an alternative specimen to blood for monitoring inflammation in COVID-19 patients.

Introduction Molecular diagnostics using RT-PCR has now emerged as the new diagnostic method for clinicians with the dawn of SARS-CoV-2. In India, the popularity and awareness of RT-PCR and particularly the increased availability of testing machines across hospitals has now opened up possibilities of diagnostic tests with RT-PCR. In view of the cytokine storm which is the significant reason for morbidity and mortality of COVID-19 patients, we proposed to test the usefulness of a multiplex RT-PCR test kit that simultaneously measured inflammatory markers namely, IL-6, TNF– α and IL-10 (IFM) all in one tube. The study included a group of patients who were equally allotted to two treatment arms one of which received standard of care along with a food supplement capsule as a natural anti-inflammatory (RECOVEREEZ FORTE™), and the other group received standard of care that included oral Prednisolone tablets in tapered dosage. RECOVEREEZ FORTE™ consists of potent biomolecules from cardamom extract. A natural product with substantial anti-inflammatory action when consumed early at the onset of symptoms is hereby proven by comparing gene expression profile of inflammatory markers with routinely tested inflammatory parameters such as serum IL-6, CRP and LDH. In addition to predicting worse disease outcomes beforehand, RT-PCR assay tests provides an opportunity for identifying therapeutic window aiding in practicing effective treatment strategy for COVID-19. RT-PCR analysis of IFM together being used in a single multiplex kit is being first reported and such a test as a prognosticator for disease progression does seem promising and worthy of clinicians adopting novel testing modalities in clinical practice. Aim of the study To study the anti-inflammatory response of RECOVEREEZ FORTE™ using RT-PCR based multiplex gene profiling of inflammatory markers in disease prognosis and show its predictability of worsening outcomes and its role in identifying therapeutic window for RECOVEREEZ FORTE™. To show the potentiality of RECOVEREEZ FORTE™ in reducing COVID-19 symptoms and SARS-CoV-2 RT-PCR Ct values of the treatment group. Materials and methods This was a short study of a 10-day period where the end point was the negativity of COVID-19 on RT-PCR test or the decline in cycle threshold (Ct) values of the test performed on day 10 when compared to day 0. During the course of the study, patients were given standard treatment and an oral dose of 500 mg of RECOVEREEZ FORTE™ thrice daily, or standard of care and oral Prednisolone in tapered dosage (control group). All essential interventions were included in the standard of care as decided by the attending physician. The RT-PCR results of inflammatory markers were compared to routinely tested parameters such as IL-6, CRP and LDH. Time to clinical improvement was in terms of SARS-CoV-2 RT-PCR test negativity or recovery of COVID-19 symptoms. We enrolled 64 patients, of which 32 were allocated to RECOVEREEZ FORTE ™ group and 32 to the control group. Results The RT-PCR analysis of elevated IFM on day 0 corresponded to above normal protein levels of routinely tested parameters such as serum IL-6, CRP and LDH on day 5. Similarly, elevated IFM on day 5 corresponded to above normal protein levels on day 10. Such an association was equally prevalent in both the control group and RECOVEREEZ FORTE™ group, stipulating that RECOVEREEZ FORTE™ may be used as an alternative to steroids. The obtained results indicate that the RT-PCR assessment predicts worse outcomes 5 days earlier. But, the RT-PCR analysis of elevated IFM on day 0 did not correspond to the above normal protein levels of other parameters on day 10. Hence, indicating that the IFM RT-PCR test cannot predict worse outcomes 10 days earlier. Also, RECOVEREEZ FORTE™ when consumed for a period of 10 days normalized LDH values, compared to the control group. Moreover, IFM RT-PCR test identified a 5-day therapeutic window for RECOVEREEZ FORTE™ against inflammation experienced by patients. In addition to the above findings, the authors also observed that majority of the patients belonging to the treatment group showed recovery from symptoms such as fever, cough, sore throat and breathlessness compared to control group by day 5. 12 (37.5%) out of 32 patients and 6 (18.75%) out of 32 patients belonging to the treatment group and control group, respectively, became SARS-CoV-2 negative by day 5, indicating a probable anti-viral action of RECOVEREEZ FORTE™ against SARS-CoV-2. Conclusion The IFM RT-PCR test possess 5-day early prediction ability and lacks 10-day prediction ability. Treatment with RECOVEREEZ FORTE™ indicates good anti-inflammatory action which is equivalent to steroids. Intake of RECOVEREEZ FORTE ™ for a period of 5 days depicts persistent anti-inflammatory action, recovery of COVID-19 symptoms and a probable anti-viral action. Moreover, an effective normalization of LDH may be rendered by RECOVEREEZ FORTE™ when consumed for a period of 10 days.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) provokes demanding immune and inflammatory events. Despite numerous reports on the use and testing of several potent therapeutic options against coronavirus disease 2019 (COVID-19), satisfactory treatment has not so far been determined. RECOVEREEZ FORTE TM , consisting of a standardized cardamom extract is a natural product with substantial indications of immunomodulatory and anti-inflammatory actions along with inhibitory capacity averse to viral targets. In this context, we speculated that RECOVEREEZ FORTE TM may ameliorate adverse effects in COVID-19 patients. Accordingly, in a multicenter prospective, open label, randomized trial, adult COVID-19 patients having mild to moderate symptoms were treated with RECOVEREEZ FORTE TM as an adjunct therapy. Patients were assigned to obtain standard of care along with a three times per day oral dose of 500 mg of RECOVEREEZ FORTE TM for ten days, or standard of care alone. Standard of care comprised all essential interventions, as per the discretion of the attending physician. Time to clinical improvement in terms of biochemical parameters such as IL6, CRP, DDIMER, LDH and an RTPCR COVID-19 test negativity in the treated patients was considered to be the primary end point. We enrolled 60 patients; of which 30 were allocated to RECOVEREEZ FORTE TM and 30 to the control group. The duration of COVID-19 positivity post-intervention was significantly shorter in RECOVEREEZ FORTE TM treated group than in the control group (p<0.001). RECOVEREEZ FORTE TM group also showed significant decrease in the levels of IL6, LDH, D-dimer along with increased lymphocytes confirming its immunomodulatory and anti-inflammatory action. We conclude that RECOVEREEZ FORTE TM can reduce the impact of COVID-19 manifestations and could serve as an alternative to oral steroids. We also suggest that RECOVEREEZ FORTE TM may aid faster recovery mediated by its immunomodulatory and anti-inflammatory action in diseases other than COVID-19. The trial was registered in Clinical Trials Registry India (CTRI - CTRI/2021/04/033143) in compliance with the International Council on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use – Good Clinical Practice (ICH–GCP) guidelines.

Naturally derived ingredients are becoming more prevalent in therapeutic drug formulations due to consumers’ concerns about chemical side effects. In the context of wound care, despite the impressive progress in therapeutic product development, drugs dispensed to treat impaired healing challenged by biofilms; excessive inflammation and oxidation are not yet really effective. Thus, the hunts for improved drug formulations preferably using natural ingredients that are cost-effective in accelerating the wound-healing process are of constant demand. The grape seed extract is extensively studied and is reported to be rich in phenolic compounds, unsaturated fatty acids and vitamins which exhibit numerous therapeutic benefits owing to their anti-inflammatory, anti-microbial, and anti-oxidative properties that support its potential use in the development of wound-healing products. We conducted a literature study using Scopus, PubMed, and Google Scholar including the keywords “grape seed extract” and “wound healing”. We also scanned all the references cited by the retrieved articles. Accordingly, this review is aimed to (i) explore the various phytochemical constituents found in grape seed extracts along with their mechanism of action that instigate wound healing, (ii) to highlight the latest pre-clinical and clinical assessments of grape seed extract in wound models, and (iii) to encourage innovation scientists in the field to address current limitations and to effectively develop grape seed extract-based wound care product formulations for commercialization.

Nowadays human saliva is more frequently studied as a non-invasive, stress-free, and preferable diagnostic material than blood. Supporting evidences acknowledge saliva as a mirror that reflects the body’s physical state. Numerous studies have also demonstrated the presence and use of RNA derived from saliva in the early diagnosis of disease by real-time reverse transcriptase polymerase chain reaction (RT-PCR). Assessing the host inflammatory response in patients and its resolution at an early stage can serve as a prognostic and predictive method in determining therapeutic response or disease progression. In this context, the potential of saliva as a specimen to diagnose early inflammatory biomarkers using RT-PCR seems fascinating and useful. Here, we review inflammatory biomarkers within the saliva, focusing on early detection of these biomarkers using RT-PCR and the factors influencing the quality of saliva specimen.