Prashanth Varkey

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.

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.

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.

Head and neck cancer remains a significant cause of morbidity worldwide. Multimodality treatment is often the only way to achieve improved function, quality of life, and survival, calling for a multidisciplinary team approach, particularly in view of the rapid advances being made in various fields. The roles of the head and neck surgeon and reconstructive surgeon are discussed, together with the input afforded by specialists in areas such as diagnostic imaging, radiation therapy, medical oncology, and gene therapy. Telemedicine is of importance in centers where multidisciplinary expertise is not available.

Anorexia and cachexia frequently complicate the late stages of malignancy and can be a prominent feature of early disease. The resulting weight loss significantly affects the morbidity and mortality of the cancer patient. A fundamental understanding of nutrition and the pathophysiology of cancer cachexia will aid in diligent treatment decisions to achieve optimal results. The pathophysiology of cancer cachexia is discussed, together with methods of nutritional assessment, nutritional requirements, and postoperative nutritional support. The advantages and disadvantages of the various modes of parenteral and enteral feeding are presented, together with information about enteral feeding in the home.

Microsurgical free tissue transfer is progressing rapidly and is being used more frequently the world over. Monitoring these cases is crucial for a satisfactory outcome. More sophisticated methods are available for monitoring, though they are often expensive. We propose a novel technique using digital photography and the Internet to offer a reliable and cost-effective method to monitor free-tissue transfers. During an 8-month period, 163 microvascular procedures were monitored using this technique. Serial photographs taken were stored in a separate case folder and sent to the surgeon as deemed necessary. Analysis of the 67 cases is presented. Five reexplorations were done. The early diagnosis of venous congestion was possible using this technique. Timely intervention contributed to the success of the reexplorations, and these flaps could be salvaged. The file size of images was in the range of 1 MB to 6 MB. The file size of a set of photographs was usually around 7 MB to 9 MB. These were sent across the asymmetric digital subscriber line Internet lines. The use of the digital images and the Internet allows reconstructive surgeons to have a reliable picture of the state of their free-tissue transfers. This allows decreasing observer error and saves valuable time which otherwise needs to be spent to verify situations of doubt.