Exploring the immunomodulatory potential of Brahmi (Bacopa monnieri) in the treatment of invasive ductal carcinoma.

Bacopa monnieri (L) Wettst, commonly known as Brahmi, stands as a medicinal plant integral to India's traditional medical system, Ayurveda, where it is recognized as a "medhya rasayana"-a botanical entity believed to enhance intellect and mental clarity. Its significant role in numerous Ayurvedic formulations designed to address conditions such as anxiety, memory loss, impaired cognition, and diminished concentration underscores its prominence. Beyond its application in cognitive health, Brahmi has historically been employed in Ayurvedic practices for the treatment of inflammatory diseases, including arthritis. In contemporary biomedical research, Bacopa monnieri can attenuate the release of pro-inflammatory cytokines TNF-α and IL-6 in animal models. However, there remains a paucity of information regarding Bacopa's potential as an anticancer agent, warranting further investigation in this domain. Based on previous findings with Brahmi (Bacopa monnieri), the current study aims to find out the role of Brahmi plant preparation (BPP) in immunomodulatory actions on IDC. Employing a specific BPP concentration, we conducted a comprehensive study using MTT assay, ELISA, DNA methylation analysis, Western blotting, ChIP, and mRNA profiling to assess BPP's immunomodulatory properties. Our research finding showed the role of BPP in augmenting the action of T helper 1 (TH1) cells which secreted interferon-γ (IFN-γ) which in turn activated cytotoxic T-lymphocytes (CTL) to kill the cells of IDC (*p < 0.05). Moreover, we found out that treatment with BPP not only increased the activities of tumor-suppressor genes (p53 and BRCA1) but also decreased the activities of oncogenes (Notch1 and DNAPKcs) in IDC (*p < 0.05). BPP had an immense significance in controlling the epigenetic dysregulation in IDC through the downregulation of Histone demethylation & Histone deacetylation and upregulation of Histone methylation and Histone acetylation (*p < 0.05). Our Chromatin immunoprecipitation (ChIP)-qPCR data showed BPP treatment increased percentage enrichment of STAT1 & BRCA1 (*p < 0.05) and decreased percentage enrichment of STAT3, STAT5 & NF ΚB (*p < 0.05) on both TBX21 and BRCA1 gene loci in IDC. In addition, BPP treatment reduced the hypermethylation of the BRCA1-associated-DNA, which is believed to be a major factor in IDC (*p < 0.05). BPP not only escalates the secretion of type 1 specific cytokines but also escalates tumor suppression and harmonizes various epigenetic regulators and transcription factors associated with Signal Transducer and Activator of Transcription (STAT) to evoke tumor protective immunity in IDC.

Wiskott Aldrich syndrome protein (WASp)-deficient Th1 cells promote R-loop-driven transcriptional insufficiency and transcription-coupled nucleotide excision repair factor (TC-NER)-driven genome-instability in the pathogenesis of T cell acute lymphoblastic leukemia.

BACKGROUND: T-ALL is an aggressive hematological tumor that develops as the result of a multi-step oncogenic process which causes expansion of hematopoietic progenitors that are primed for T cell development to undergo malignant transformation and growth. Even though first-line therapy has a significant response rate, 40% of adult patients and 20% of pediatric patients will relapse. Therefore, there is an unmet need for treatment for relapsed/refractory T-ALL to develop potential targeted therapies. METHODS: Pediatric T-ALL patient derived T cells were grown under either nonskewingTh0 or Th1-skewing conditions to further process for ChIP-qPCR, RDIP-qPCR and other RT-PCR assays. Endogenous WASp was knocked out using CRISPR-Cas9 and was confirmed using flow cytometry and western blotting. LC-MS/MS was performed to find out proteomic dataset of WASp-interactors generated from Th1-skewed, human primary Th-cells. DNA-damage was assessed by immunofluorescence confocal-imaging and single-cell gel electrophoresis (comet assay). Overexpression of RNaseH1 was also done to restore normal Th1-transcription in WASp-deficient Th1-skewed cells. RESULTS: We discovered that nuclear-WASp is required for suppressing R-loop production (RNA/DNA-hybrids) at Th1-network genes by ribonucleaseH2 (RNH2) and topoisomerase1. Nuclear-WASp is associated with the factors involved in preventing and dissolving R-loops in Th1 cells. In nuclear- WASp-reduced malignant Th1-cells, R-loops accumulate in vivo and are processed into DNA-breaks by transcription-coupled-nucleotide-excision repair (TC-NER). Several epigenetic modifications were also found to be involved at Th1 gene locus which are responsible for active/repressive marks of particular genes. By demonstrating WASp as a physiologic regulator of programmed versus unprogrammed R-loops, we suggest that the transcriptional role of WASp in vivo extends also to prevent transcription-linked DNA damage during malignancy and through modification of epigenetic dysregulations. CONCLUSION: Our findings present a provocative possibility of resetting R-loops as a therapeutic intervention to correct both immune deficiency and malignancy in T-cell acute lymphoblastic leukemia patients and a novel role of WASp in the epigenetic regulation of T helper cell differentiation in T-ALL patients, anticipating WASp's requirement for the suppression of T-ALL progression.

Immunomodulatory effects of Diospyros peregrina fruit preparation (DFP) in non-small cell lung cancer (NSCLC) by utilizing dendritic cell-mediated antigen presentation and T helper (TH) cell differentiation.

Diospyros peregrina is a dioecious plant which is native to India. It belongs to the family of Ebenaceae and is extensively used to treat various ailments, such as leucorrhoea and other uterine-related problems. Though few studies have been on D. peregrina for their anti-tumour response, little is known. Therefore, this intrigued us to understand its immunomodulator capabilities on various types of cancer extensively. Our primary focus is on NSCLC (Non-Small Cell Lung Cancer), which is ranked as the second largest form of cancer in the world, and the treatments demand non-invasive agents to target NSCLC effectively. In an objective to generate an efficient Lung Cancer Associated Antigen (LCA) specific anti-tumour immune response, LCA was presented using dendritic cells (DCs) in the presence of D. peregrina fruit preparation (DFP). Moreover, we also investigated DFP's role in the differentiation of T-helper (TH) cells. Therefore, this study aimed at better LCA presentation mediated by DFP by activating the LCA pulsed DCs and T helper cell differentiation for better immune response. DCs were pulsed with LCA for tumour antigen presentation in vitro, with and without DFP. Differentially pulsed DCs were irradiated to co-culture with autologous and allogeneic lymphocytes. Extracellular supernatants were collected for the estimation of cytokine levels by ELISA. LDH release assay was performed to test Cytotoxic T lymphocytes (CTLs) mediated lung tumour cell cytotoxicity. Thus, DFP may be a potential vaccine to generate anti-LCA immune responses to restrict NSCLC.

Comparison of Efficacy of Platelet-Rich Plasma With and Without Topical Minoxidil for Hair Growth in Patients With Androgenetic Alopecia: A Prospective Study.

Introduction Androgenetic pattern of alopecia is a common problem occurring in men, which mostly arises from their younger age. There are many therapies advocated in the literature for hair loss reduction, and one of them is platelet-rich plasma (PRP) therapy. This study aimed to assess the efficacy of combined PRP therapy with topical minoxidil over PRP as monotherapy in hair loss reduction and regeneration of new hair. Materials and methods The study was conducted at our institute in the Department of Oral and Maxillofacial Surgery at Saveetha Dental College and Hospital. The study consisted of 40 participants, 20 of whom had only PRP therapy as part of their treatment, while the other 20 participants received PRP combined with topical minoxidil as treatment. Both group participants were evaluated for postoperative hair shaft diameter and hair follicle density. Parameters were measured preoperatively and postoperatively after one month, two months, and three months. Data analysis was done with the help of SPSS, with P-values less than 0.05 considered statistically significant. The Mann-Whitney U test was used to compare the two groups for measurement of hair shaft diameter, and for comparison between hair follicle density, an unpaired t-test was used. Results It was found that the mean hair shaft diameter in the PRP with minoxidil group was higher than that of the PRP group for one month (P = 0.023), two months (P = 0.001), and three months (P = 0.001) postoperative periods, and the results were statistically significant. Hair follicle density (mean hair quantity) was higher in the PRP group than in the PRP with the minoxidil group in the first postoperative month. However, this difference was not statistically significant (P = 0.08). While the mean hair quantity in the PRP with minoxidil group was higher than that in the PRP group for two months (P = 0.45) and three months (P = 0.001) postoperative periods, the results were statistically significant only at the three-month postoperative period. Conclusion It can be concluded that injectable autologous PRP with minoxidil as a topical agent is a better treatment option for the improvement of both hair quality (hair shaft diameter) and hair quantity (hair follicle density) compared to plain autologous injectable PRP monotherapy.

Doxorubicin induced epigenetic regulation of dendritic cell maturation in association with T cell activation facilitates tumor protective immune response in non-small cell lung cancer (NSCLC).

BACKGROUND: NSCLC is one of the leading causes of death and is often diagnosed at late stages with no alternative therapeutic approach. DCs are professional antigen-presenting cells and DC-based immunotherapy has been under the spotlight for its anti-cancer properties. Epigenetic modifications including DNA methylation and histone modification in DCs play a crucial role in regulating their functions such as maturation and activation,innate immune responses, T cell priming, antigen presentation, and cytokine production. In the current study, we investigated the anti-cancer properties of Doxorubicin at a noncytotoxic concentration that could be extrapolated as an epigenetic regulator for DC maturation to elicit anti-tumor activity. METHODOLOGIES: PBMCs from normal and NSCLC blood samples were isolated and treated with growth factors. DCs were matured with low dose Doxorubicin and the DC maturation markers were checked by using flow-cytometry. Further, ELISA was performed and low dose Doxorubicin-induced DCs were pulsed with LCA (Lung Cancer Antigen) and primed with CD4 +T helper (Th) cells for cytotoxicity assessment. Further, epigenetic markers of T: DC conjugation were immunofluorescently visualized under a microscope. ChIP-qPCR and Invitro assays such as histone methylation, DNA methylation, and m6A methylation were performed to study the epigenetic changes under low dose Dox treatment. IL-12 neutralization assay was performed to check for the IL-12 dependency of DCs and their effect under Dox at low dose treatment. This was further followed by a Western Blotting analysis for histone and non-histone proteins. RESULTS: Low dose Doxorubicin induces epigenetic changes in DCs to elicit an anti-tumor response in NSCLC through the generation of CTLs with a concomitant increase in the extracellular secretions of anti-inflammatory cytokines. We also found that low dosage of Doxorubicin matured DCs when pulsed with LCA and primed with CD4 +T helper cells, secrete IFN-γ which is important in orchestrating adaptive immunity by activating CD8 + cytotoxic T-lymphocytes. Also, the secretions of IL-12 help us infer that protective immunity is also induced via Th1 response which triggered selectively the translocation of PKCθ to immunological synapse in between DC and Th. Further, methylation and acetylation markers H3K4me3 and H3K14Ac respectively upregulated whereas levels of STAT5, NFkB, NOTCH1, and DNAPKcs were downregulated. DNA and RNA methylation assays then lead to confirmations about the epigenetic changes caused by low dose Dox treatment. DNA methylation was reduced which resulted in the activation of tumor suppressor gene p53 and Th1-associated transcription factor TBX21. On the other hand, both absolute and relative RNA methylation quantification increased in the presence of Dox at a low dose. CONCLUSION: From this study, we understand that non-cytotoxic concentration of Doxorubicin increases the Ag-presenting ability of DCs via an IL-12-dependent mechanism and causes epigenetic modifications in NSCLC.

Association of Wiskott-Aldrich syndrome protein (WASp) in epigenetic regulation of B cell differentiation in non-small-cell lung cancer (NSCLC).

Non-small-cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer which is the deadliest type of cancer for both men and women. Previous studies already showed that cell-intrinsic loss of WASp causes B cell tolerance and WASp deficiency in T helper (TH) cells is linked to negative effects on cytokine gene transcription necessary for TH1 differentiation. In the current study, we investigated the molecular mechanisms involved in WASp-mediated epigenetic regulation of B cell differentiation during NSCLC. Our ChIP-qPCR data suggest the less percentage enrichment of the B cell differentiating factors (Ikaros, Pax5, PU.1, BATF) and WASp across the WAS gene in the B cells of NSCLC patients in comparison with normal healthy donors and overexpression of WASp showed the reverse effects. WASp-depleted B cells while co-culturing with respective PBMCs isolated from normal healthy donors and NSCLC patients, we observed upregulation of TH2-, TH17-, and Treg-specific cytokines (IL4, ILI7A, IL10) & transcription factors (GATA3, RORC, FOXP3) and downregulation of TH1-specific cytokine (IFNγ) & transcription factor (TBX21). Our study showed that the overexpression of WASp resulted into upregulation of B cell differentiating factors, tumor suppressor protein (p53), histone methylation marker (H3K4me3) with concomitant downregulation of tumor-promoting factors (Notch 1, ß-Catenin, DNAPKcs) and histone deacetylation marker (HDAC2) and increase in percentage cytotoxicity of NSCLC-specific cells (A549). Successful overexpression of WASp not only helps in epigenetic regulation of B cell differentiation but also supports tumor suppression in NSCLC. Thus, WASp can be targeted for therapeutic intervention of NSCLC.

Efficacy of Dexamethasone Diluted Saline Irrigant on Postoperative Sequelae in Patients Undergoing Lower Third Molar Surgery: A Prospective Clinical Study.

Background Third molar impaction surgery is one of the most common yet challenging procedures done as a part of minor oral surgery. Years of research and improvisation of techniques have been done, yet there are still a lot of postoperative sequelae after surgical removal of the impacted tooth. In our study, we have compared the efficacy of dexamethasone diluted saline solution over plain saline solution used as an irrigant in the reduction of postoperative sequelae for lower third molar surgery. Aim The aim of the study was to evaluate the efficacy of dexamethasone diluted saline solution over plain saline solution in the reduction of postoperative sequelae for lower third molar surgery. Materials and methods The research was conducted at Saveetha Dental College and Hospital in the Department of Oral and Maxillofacial Surgery. The study consisted of 48 individuals, 24 of whom had dexamethasone saline as an irrigant (8 mg of dexamethasone was diluted in 100 ml of plain saline) (Group 1), and 24 in whom plain saline was used as an irrigant (Group 2) in the lower third molar surgery. Patients were evaluated postoperatively for pain and swelling. The postoperative swelling was measured on postoperative day two and day seven. Postoperative pain was measured on day two, day four, and day seven after surgery using a visual analog scale. Data were analyzed using SPSS (IBM Corp., Armonk, NY) with P-values less than 0.05 considered statistically significant. The statistical test used to compare the outcomes between the two groups was the independent samples t-test. Results It was found that study participants in the dexamethasone saline irrigation group reported statistically significantly lesser pain than participants receiving plain saline irrigation on day two (P = 0.001), day four (P = 0.001), and day seven (P = 0.001), respectively. Also, there was a reduction in swelling among participants in the dexamethasone saline irrigation group when compared to the normal saline irrigation group, which was statistically significant (P = 0.001) on day two, while the postoperative swelling was not statistically significant on day seven (P = 0.08) between the two study groups. Conclusion Based on the results obtained, it can be concluded that dexamethasone saline solution (8 mg/100 mL) was more effective as an irrigant in reducing the postoperative sequelae than regular saline solution in the lower third molar surgery.

Preemptive Analgesic Efficacy of Dexamethasone and Diclofenac in Mitigating Post-surgical Complications After Mandibular Third-Molar Surgery: A Systematic Review.

Mandibular third-molar extraction is a frequently executed minor oral surgical procedure, with a subsequent recovery period lasting several days. Typically, preemptive administration of non-steroid anti-inflammatory drugs (NSAIDs) and steroids has been employed, resulting in a notable decrease in postoperative complications like pain, facial swelling, trismus, and alveolar osteitis. This systematic review's primary goal was to investigate the efficacy of preemptive analgesia with dexamethasone and diclofenac in minimizing the post-surgical complications following the surgical extraction of the mandibular third molars. The systematic search was carried out to identify relevant literature in digital databases including PubMed®, Cochrane Library, Web of Science, and Scopus, from January 1990 to January 2022. The search used specific keywords. The randomized clinical trials assessing the efficacy of dexamethasone and diclofenac or dexamethasone alone compared to diclofenac or placebo as preemptive analgesics were considered inclusion criteria for this systematic review. Case reports, literature reviews, letters to the editor, and non-English publications were not included. Two authors screened the titles and abstracts, and articles fulfilling the study criteria were included. After reading the full text and data collection, analysis was performed. The included article's bias was evaluated by the Risk of Bias 2 (RoB 2) tool. A digital database search yielded a total of 207 articles. After excluding duplicates and articles written in languages other than English, 90 were removed. Based on the title and abstract, out of 177, 95 studies were excluded. After full-text reading of 22 articles, 17 were eliminated because they did not meet the inclusion and exclusion criteria. The remaining five studies were found eligible and included in the systematic review. Four studies were of low risk, while one study had some concerns. Two studies evaluated the combination of dexamethasone with diclofenac, while three evaluated dexamethasone alone. Total samples included samples of 436 third-molar surgeries in 420 patients. There was a substantial decrease in the mean pain score and swelling measurement when diclofenac alone was compared with coadministration of diclofenac and dexamethasone. Preemptive administration of dexamethasone and diclofenac has been shown to effectively reduce pain and facial swelling, with the exception of trismus, in third-molar surgeries when compared to diclofenac alone. As a result, it is recommended to administer these drugs prior to the commencement of third-molar extraction. However, further research is mandatory, specifically good quality randomized controlled trials involving large cohorts, in order to assess any significant variations and validate these findings.

Prediction of maternal and foetal outcomes among patients with preeclampsia using circulatory biomarkers (MMP-9 & ST2) - A prospective cohort study.

INTRODUCTION: Preeclampsia is one of the major causes of maternal and foetal morbidity and mortality worldwide. Its complications include but are not limited to eclampsia, intracerebral haemorrhage and cardiovascular diseases in the later stages of life. The combination of clinical and risk variables and a panel of multiple biomarkers will help clinicians in risk stratification and prognostication of clinical outcomes among preeclamptic women. We evaluated MMP-9 (matrix metalloproteinase - 9) and ST2 (suppression of tumorigenicity 2) for utility as biomarkers and for predicting maternal and foetal outcomes in women with preeclampsia. METHODS: This prospective cohort study involved 49 preeclamptic women and 80 healthy controls. Biomarkers were measured in plasma using ELISA. The patients were followed up to assess maternal and foetal outcomes. RESULTS: The mean value of MMP-9 was 2.42 ng/mL in the preeclamptic group and 2.67 ng/mL in controls. The mean value of ST2 (1937.4 ± 747.81) in the preeclamptic group was high compared to the control group (1005.7 ± 683.6) and the difference was significant (P = 0.0001). The study population was divided into those with high and low MMP-9 and those with high and low ST2. Lower levels of MMP-9 seemed to be related to both early and late onset preeclampsia. The ROC (Receiver Operating Characteristic) curve did not show the ability to predict maternal and foetal outcomes. DISCUSSION: Our study demonstrated that women with preeclampsia had low MMP-9 and high ST2 compared to healthy pregnant women. But neither of the biomarkers could predict complications of preeclampsia.

Segmental Resection With Primary Reconstruction Using a Patient-Specific Implant for Unicystic Ameloblastoma: A One-Year Follow-Up Case Report.

Unicystic ameloblastoma is a slow-growing tumor originating from the odontogenic epithelium that can be localized within the lining of a cyst. It commonly affects younger individuals and is frequently found in the posterior mandible. The classification of this tumor is based on histopathological characteristics, distinguishing between the luminal, intraluminal, and mural proliferation of the odontogenic epithelium. Treatment options vary depending on the histology and can range from enucleation to resection with secondary reconstruction. In recent years, patient-specific implants have gained popularity in reconstructive surgeries, particularly in craniomaxillofacial surgery. This case report focuses on a 22-year-old female patient with a mural-type unicystic ameloblastoma. The treatment involved segmental mandibular resection with primary reconstruction using a patient-specific implant to address the mandibular defect. The postoperative healing process and condylar movement were evaluated, and the patient achieved satisfactory results. This case report provides valuable insights into the management of primary reconstruction using a patient-specific implant.

HDAC inhibition regulates oxidative stress in CD4+Thelper cells of chronic obstructive pulmonary disease and non-small cell lung cancer patients via mitochondrial transcription factor a (mtTFA) modulating NF-κB/HIF1α axis.

Histone deacetylases (HDACs) play a crucial role in the epigenetic regulation of gene expression by remodelling chromatin. Isoenzymes of the HDAC family exhibit aberrant regulation in a wide variety of cancers as well as several inflammatory lung disorders like chronic obstructive pulmonary disease (COPD). Inhibition of HDACs is a potential therapeutic strategy that could be used to reverse epigenetic modification. Trichostatin A (TSA), a powerful histone deacetylase (HDAC) inhibitor, has anti-cancer effects in numerous cancer types. However, it is not yet apparent how HDAC inhibitors affect human non-small cell lung cancer cells (NSCLC) and COPD. This study aims to investigate TSA's role in restoring mitochondrial dysfunction and its effect on hypoxia and inflammation in CD4+T cells obtained from patients with COPD and lung cancer. As a result of treatment with TSA, there is a reduction in the expression of inflammatory cytokines and a decreased enrichment of transcriptional factors associated with inflammation at VEGFA gene loci. We have seen a substantial decrease in the expression of NF-κB and HIF1α, which are the critical mediators of inflammation and hypoxia, respectively. Following TSA treatment, mtTFA expression was increased, facilitating patients with COPD and NSCLC in the recovery of their dysfunctional mitochondria. Furthermore, we have discovered that TSA treatment in patients with COPD and NSCLC may lead to immunoprotective ness by inducing Th1ness. Our finding gives a new insight into the existing body of knowledge regarding TSA-based therapeutic methods and highlights the necessity of epigenetic therapy for these devastating lung disorders.

Genome-wide identification and expression analysis of the polygalacturonase gene family in sweetpotato.

BACKGROUND: Polygalacturonase (PG), a crucial enzyme involved in pectin degradation, is associated with various plants' developmental and physiological processes such as seed germination, fruit ripening, fruit softening and plant organ abscission. However, the members of PG gene family in sweetpotato (Ipomoea batatas) have not been extensively identified. RESULTS: In this study, there were 103 PG genes identified in sweetpotato genome, which were phylogenetically clustered into divergent six clades. The gene structure characteristics of each clade were basically conserved. Subsequently, we renamed these PGs according to their locations of the chromosomes. The investigation of collinearity between the PGs in sweetpotato and other four species, contained Arabidopsis thaliana, Solanum lycopersicum, Malus domestica and Ziziphus jujuba, revealed important clues about the potential evolution of the PG family in sweetpotato. Gene duplication analysis showed that IbPGs with collinearity relationships were all derived from segmental duplications, and these genes were under purifying selection. In addition, each promoter region of IbPG proteins contained cis-acting elements related to plant growth and development processes, environmental stress responses and hormone responses. Furthermore, the 103 IbPGs were differentially expressed in various tissues (leaf, stem, proximal end, distal end, root body, root stalk, initiative storage root and fibrous root) and under different abiotic stresses (salt, drought, cold, SA, MeJa and ABA treatment). IbPG038 and IbPG039 were down-regulated with salt, SA and MeJa treatment. According to the further investigation, we found that IbPG006, IbPG034 and IbPG099 had different patterns under the drought and salt stress in fibrous root of sweetpotato, which provided insights into functional differences among these genes. CONCLUSION: A total of 103 IbPGs were identified and classified into six clades from sweetpotato genome. The results of RNA-Seq and qRT-PCR suggested that IbPG006, IbPG034 and IbPG099 might play a significant role in tissue specificity as well as drought and salt stress responses, which showed valuable information for further functional characterization and application of the IbPGs.

Diospyros malabarica fruit preparation mediates immunotherapeutic modulation and epigenetic regulation to evoke protection against non-small cell lung cancer (NSCLC).

ETHNOPHARMACOLOGICAL RELEVANCE: Diospyros malabarica is an ethnomedicinal plant with hypoglycaemic, anti-bacterial, and anti-cancer properties and it belongs to the Ebenaceae family which is well known for its medicinal uses since ancient times and application of its bark and unripened fruit has been significantly mentioned in Ayurvedic texts. The Diospyros malabarica species which is known as the Gaub in Hindi and Indian Persimmon in English is native to India, however, it is distributed throughout the tropics. AIM OF THE STUDY: As Diospyros malabarica fruit preparation (DFP) possesses medicinal values, the study aims to evaluate its role as natural, non-toxic, and cost-effective dendritic cells (DCs) maturing immunomodulatory agent and also as an epigenetic regulator to combat Non-small cell lung cancer (NSCLC) which is a type of lung cancer whose treatment options such as chemotherapy, radiation therapy, etc. are accompanied with some adverse side effects. Thus, immunotherapeutic strategies are in high demand to evoke tumor protective immunity against NSCLC without causing such side effects. MATERIALS AND METHODS: Peripheral Mononuclear Cells (PBMCs) derived monocytes of normal subjects and NSCLC patients were utilized to generate DCs matured with either LPS (LPSDC) or DFP (DFPDC). Mixed Lymphocyte Reaction (MLR) was carried out with the differentially matured DCs co-culturing T cells and cytotoxicity of lung cancer cells (A549) was measured through LDH release assay and cytokine profiling was carried out via ELISA respectively. PBMCs of normal subjects and NSCLC patients have transfected separately in vitrowith CRISPR-activation plasmid of p53 and CRISPR-Cas9 knockout plasmid of c-Myc to analyze epigenetic mechanism(s) in the presence and absence of DFP. RESULTS: Diospyros malabarica fruit preparation (DFP) treated DC upregulates the secretion of T helper (TH)1 cell specific cytokines (IFN-γ and IL-12) and signal transducer and activator of transcription molecules (STAT1 and STAT4). Furthermore, it also downregulates the secretion of TH2-specific cytokines (IL-4 and IL-10). Diospyros malabarica fruit preparation (DFP) enhances p53 expression by reducing methylation levels at the CpG island of the promoter region. Upon c-Myc knockout, epigenetic markers such as H3K4Me3, p53, H3K14Ac, BRCA1, and WASp were enhanced whereas H3K27Me3, JMJD3, and NOTCH1 were downregulated. CONCLUSION: Diospyros malabarica fruit preparation (DFP) not only increases the expression of type 1 specific cytokines but also augments tumor suppression modulating various epigenetic markers to evoke tumor protective immunity without any toxic activities.

Depletion of enhancer zeste homolog 2 (EZH2) directs transcription factors associated with T cell differentiation through epigenetic regulation of Yin Yang 1(YY1) in combating non-small cell lung cancer (NSCLC).

Non-Small Cell Lung Cancer (NSCLC) is the leading cause of death in all countries alike. In the current study, we have found out that Histone H3Lys4trimethylation is abnormal on YY1 in CD4+T Helper (TH) cells of NSCLC patients which is evident by Histone H3Lys27 trimethylation mediated via EZH2. We investigated the status of Yin Yang 1 (YY1) and the involvement of certain transcription factors that lead to tumorigenesis after depleting endogenous EZH2 in vitro by CRISPR/Cas9 in the CD4+TH1-or-TH2-polarized cells isolated initially as CD4+TH0 cells from the PBMC of the control subjects and patients suffering from NSCLC. After depletion of endogenous EZH2, RT-qPCR based mRNA expression analysis showed that there was an increase in the expression of TH1 specific genes and a decrease in the expression of TH2 specific genes in NSCLC patients CD4+TH cells. We can conclude that this group of NSCLC patients may have the tendency at least in vitro to elucidate adaptive/protective immunity through the depletion of endogenous EZH2 along with the reduction in the expression of YY1. Moreover, depletion of EZH2 not only suppressed the CD4+CD25+FOXP3+Regulatory T cells (Treg) but also it aided the generation of CD8+Cytotoxic T Lymphocytes (CTL) which were involved in killing of the NSCLC cells. Thus the transcription factors involved in EZH2 mediated T cell differentiation linked to malignancies offers us an appealing avenue of targeted therapeutic intervention for NSCLC.

Evaluation of Endpoints Used in Clinical Trials on Adenomyosis - A Systematic Review.

Adenomyosis lacks approved pharmacological treatment even after decades of its identification. We performed this study to review the status of clinical research on adenomyosis for finding an effective drug therapy and to identify the most common endpoints used in adenomyosis trials. A systematic search was performed in the PubMed and Clinicaltrials.gov registries to identify interventional trials for analysis without any time and language restrictions. Our search revealed that barely 15 drugs have been assessed for the management of adenomyosis from 2001 to 2021. Among these, LNG-IUS was found to be the most evaluated drug, followed by dienogest. In these trials, the most commonly assessed endpoints included VAS, NPRS for pain, haemoglobin and PBAC for menstrual bleeding, uterine volume, and serum estradiol. There appears to be a need for developing a comprehensive score that takes into consideration all disease symptoms as well as incorporates some objective elements to evaluate the disease.

Differential Expression of Caveolin-3, Suppression of Tumorigenicity 2, and Growth Differentiation Factor-15 Genes and Their Association with Acute Myocardial Infarction: A Cross-Sectional Study in a Multi-Specialty Hospital in Tamil Nadu.

Background: Acute myocardial infarction (AMI) is one of the world's leading causes of cardiovascular death. Recent studies have reported the influence of the genes caveolin-3 (CAV3), suppression of tumorigenicity 2, and growth differentiating factor-15 in cardiovascular diseases, especially myocardial infarction, but their role and function remain unclear. Hence, this study was designed to evaluate the expression levels of these three genes in AMI and understanding the role of CAV-3 in the pathogenesis of AMI. Methods and Results: Blood samples were collected from 50 AMI patients and 50 non-AMI controls in this cross-sectional study. Relative expression levels of the three genes were performed using real-time PCR. Bioinformatics tools were used for functional gene enrichment and protein-protein interactions. CAV-3 was significantly upregulated among AMI patients compared to controls. In silico analyses identified CAV-3 as playing critical roles in smooth muscle contraction, cardiac conduction, and calcium-mediated transport via binding with essential proteins including dysferlin and annexins Conclusion: This study is a first of its kind, reporting an upregulation of CAV-3 in AMI patients. The expression of all three genes significantly influenced the systolic function of the heart in AMI patients. A more in-depth understanding of CAV-3 in the pathophysiology of AMI is essential and it may prove to be a novel.

Immunomodulatory Role of Diospyros peregrina Fruit Preparation in Breast Cancer by Utilizing Macrophage Mediated Antigen Presentation and T Helper Cell (Th) Differentiation.

BACKGROUND: Diospyros peregrina is dioecious plant native to India and belonging to the family of Ebenaceae, is largely utilized in treatment of various ailments. Little has been known about the antitumor activity of Diospyros peregrina with only 1 previous study on Ehrlich Ascites Carcinoma in mice. Therefore, it prompted us to extensively explore the immunomodulatory effect in various cancer forms. The focal point of this study revolves around breast cancer, which is the second most common cancer in the world. In view of the increasing demands for noninvasive treatments, natural plant-based agents open up promising applications in cancer immunotherapy METHODS: CD4+ lymphocytes were isolated from the peripheral blood mononuclear cells (PBMCs) of breast cancer patients and normal donor blood samples using magnetic-activated cell sorting (MACS) and cultured separately. Utilizing the plastic surface adherence property, the macrophages were isolated from CD4 negative lymphocytes of both breast cancer patients and normal donors. For the presentation of tumor antigens invitro, macrophages were pulsed with breast tumor associated antigen (BTAA) in presence or absence of Diospyros peregrina fruit preparation (DFP). Differentially pulsed and irradiated macrophages were co-cultured with autologous and allogenic lymphocytes. Supernatants hence collected from CD4+ lymphocytes were utilized for cytokine profiling using ELISA and proliferation was assessed by MTT assay. Cytotoxic T lymphocytes (CTLs) generated from CD4 negative lymphocytes culture (2 × 105) was incubated with MCF-7 (2 × 104) to check cytotoxicity using LDH release assay. CD4+ lymphocytes were treated in presence or absence of DFP, were analyzed using immunoblotting and RT-qPCR, to check DFP mediated T helper (Th) cell differentiation through investigation of signatory cytokines and transcription factors. RESULTS: It was found that DFP elevated the proliferation of CD4+ T lymphocytes (Th) in response to BTAA. DFP also helped in presenting BTAA pulsed macrophages directing in the cytotoxic T-lymphocyte mediated immune response. Results indicated that DFP preferentially highlighted Th1 commitment with type-1 specific cytokines IFN-g and IL-12 and was indifferent in Th2 manifestation. DFP was not only involved in the upregulation of Tbet mounted type-1 mediated immune response and activation of STAT1 but also it downregulated STAT6 and GATA3, the functional activators and regulators of type-2 immune response. Moreover, it was observed that DFP inhibited the tumor-promoting environment modulated through Tregs by downregulating Foxp3 and STAT5. Further, it was detected that DFP directs Th1 bias and results in attainment of better suppression of breast tumor CONCLUSION: The results collectively pointed out that DFP favored cell-mediated immune response from BTAA antigen presentation on macrophages and also helping in the robust proliferation of an entire spectrum of T helper lymphocytes which furthermore strengthen the underlying immune responses, hence, fencing the body, of the progression of breast cancer.

Genome-Wide Identification and Expression Analysis of the Xyloglucan Endotransglucosylase/Hydrolase Gene Family in Sweet Potato [Ipomoea batatas (L.) Lam].

The xyloglucan endotransglucosylase/hydrolase (XET/XEH, also named XTH) family is a multigene family, the function of which plays a significant role in cell-wall rebuilding and stress tolerance in plants. However, the specific traits of the XTH gene family members and their expression pattern in different tissues and under stress have not been carried out in sweet potato. Thirty-six XTH genes were identified in I. batatas, all of which had conserved structures (Glyco_hydro_16). Based on Neighbor-Joining phylogenetic analysis the IbXTHs can be divided into three subfamilies-the I/II, IIIA, and IIIB subfamilies, which were unevenly distributed on 13 chromosomes, with the exception of Chr9 and Chr15. Multiple cis-acting regions related to growth and development, as well as stress responses, may be found in the IbXTH gene promoters. The segmental duplication occurrences greatly aided the evolution of IbXTHs. The results of a collinearity analysis showed that the XTH genes of sweet potato shared evolutionary history with three additional species, including A. thaliana, G. max, and O. sativa. Additionally, based on the transcriptome sequencing data, the results revealed that the IbXTHs have different expression patterns in leaves, stems, the root body (RB), the distal end (DE), the root stock (RS), the proximal end (PE), the initiative storage root (ISR), and the fibrous root (FR), and many of them are well expressed in the roots. Differentially expressed gene (DEG) analysis of FRs after hormone treatment of the roots indicated that IbXTH28 and IbXTH30 are up-regulated under salicylic acid (SA) treatment but down-regulated under methyl jasmonate (MeJA) treatment. Attentionally, there were only two genes showing down-regulation under the cold and drought treatment. Collectively, all of the findings suggested that genes from the XTH family are crucial for root specificity. This study could provide a theoretical basis for further research on the molecular function of sweet potato XTH genes.

Effect of Heat Treatment on the Quality and Soft Rot Resistance of Sweet Potato during Long-Term Storage.

Heat treatment is a widely applied technique in the preservation of fruits and vegetables, effectively addressing issues such as disease management, rot prevention, and browning. In this study, we investigated the impact of heat treatment at 35 °C for 24 h on the quality characteristics and disease resistance of two sweet potato varieties, P32/P (Ipomoea batatas (L.) Lam. cv 'Pushu13') and Xinxiang (Ipomoea batatas (L.) Lam. cv 'Xinxiang'). The growth in vitro and reproduction of Rhizopus stolonifer were significantly inhibited at 35 °C. However, it resumed when returned to suitable growth conditions. The heat treatment (at 35 °C for 24 h) was found to mitigate nutrient loss during storage while enhancing the structural characteristics and free radical scavenging capacity of sweet potato. Additionally, it led to increased enzyme activities for APX, PPO, and POD, alongside decreased activities for Cx and PG, thereby enhancing the disease resistance of sweet potato against soft rot. As a result, the heat treatment provided a theoretical basis for the prevention of sweet potato soft rot and had guiding significance for improving the resistance against sweet potato soft rot.