Solid and liquid state characterization of tetrahydrocurcumin using XRPD, FT-IR, DSC, TGA, LC-MS, GC-MS, and NMR and its biological activities.

Tetrahydrocurcumin (THC) is one of the major metabolites of curcumin (CUR), an ancient bioactive natural polyphenolic compound. This research article describes both the solid and liquid state characterization of THC using advanced spectroscopic and thermo-analytical techniques. Anti-inflammatory, anti-oxidant, and neuroprotective activities of THC were investigated using in vitro cell lines. Liquid chromatography-mass spectrometry analysis revealed that our sample comprised 95.15% THC, 0.51% tetrahydrodemethoxycurcumin (THDC), 3.40% hexahydrocurcumin, and 0.94% octahydrocurcumin. Gas chromatography-mass spectrometry analysis indicated the presence of 96.68% THC and 3.32% THDC. THC in solution existed as keto-enol tautomers in three different forms at different retention time, but the enol form was found to be dominant, which was also supported by nuclear magnetic resonance analysis. THC was thermally stable up to 335.55 °C. THC exhibited more suppression of cytokines (TNF-α, IL-1ß, and MIP-1α) than CUR in a concentration-dependent manner in mouse splenocytes, while NK-cell and phagocytosis activity was increased in macrophages. THC showed a significant reduction of free radicals (LPO) along with improved antioxidant enzymes (SOD and catalase) and increased free radical scavenging activity against ABTS+ radicals in HepG2 cells. THC displayed higher protection capability than CUR from oxidative stress and neuronal damage by improving cell viability against H2O2 induced HepG2 cells and MPP+ induced SH-SY5Y cells, respectively, in a concentration-dependent manner. Thus, a variation of the biological activities of THC might rely on its keto-enol form and the presence of other THC analogs as impurities. The present study could be advantageous for further research on THC for better understanding its physicochemical properties and biological variation.

Role of Vital Trace Elements in Nanocurcumin-Centered Formulation: A Novel Approach to Resuscitate the Immune System.

The present paper described the immunomodulatory potential of novel nanocurcumin-based formulation enriched with trace elements and vitamins on cyclophosphamide-induced immunosuppression in rat model. Major immune-related assays were monitored such as hemagglutination assay, delayed-type hypersensitivity (DTH) reaction, cellular immune response, IgG, IgE, IgM, cerebrospinal fluid biomarkers, hematological study, antioxidant profile, and lipid biomarkers. Chemical characterization of novel formulation showed retention time (R t ) 18.98 of curcumin, while LC-MS data revealed the presence of the curcumin mass at m/z 369.01 [M + H]+ (calculated for C21H21O6+, 369.13). This novel formulation exhibited significantly (p ≤ 0.001) increased primary and secondary antibody titer by 72.41% and 33.25%, respectively, while DTH response being improved by 87.50% (p ≤ 0.01). However, CD4+, CD8+, and CD28+ counts were significantly (p ≤ 0.05) increased by 76.46%, 68.21%, and 19.29%, respectively, while the concentrations of IgE, IgM, and IgG were significantly (p ≤ 0.05) increased by 40%, 28.43%, and 38.75%, respectively. CSF biomarkers analysis showed a decreased level of corticosterone, dopamine, serotonin, and tau protein by 29.38%, 51.73%, 29.93%, and 4.87%, respectively. Antioxidant enzymes such as CAT, GPx, and SOD were increased by 43.74%, 49.00%, and 40.84%, respectively, and non-enzymatic component, GSH, was increased by 55.52%. Similarly, free radical LPO was significantly (p ≤ 0.05) decreased by 40.37%, and acute inflammatory marker, MPO concentration, was reduced by 31.14%, compared with the disease control group. In addition, supportive hematology and lipid profile analysis showed promising results with improved overall animal profile. Thus, trace elements in novel formulation can be used in the various pharmacological activities and as dietary supplement due to its wide properties.

Immunomodulatory potential of nanocurcumin-based formulation.

Vitamins, minerals, and nanocurcumin play a substantial role in various nutraceutical/pharmaceutical formulations that are widely used in therapeutics, cosmetics, and dietary supplements. The current study aimed to investigate the comparative in vitro immunomodulatory effect of a novel nanocurcumin-based formulation with curcumin in LPS-induced cytokine expression, NK cells' activity, and phagocytosis. The proinflammatory cytokines (TNF-α, IL-1ß, and MIP-1α) and NK cells' activity were measured in cell supernatants using ELISA assay; however, phagocytosis activity was performed using colorimetric analysis. The chemical characterization of novel nanocurcumin-based formulation using LC-MS (R t 19.02 min) and mass spectra analysis (m/z 369.04) confirmed the presence of the curcumin in highest peak concentration. MTT assay in three tested cell-lines showed that the formulation was found non-toxic at all the tested concentrations. The expression of TNF-α, IL-1ß, and MIP-1α in splenocytes was significantly (p ≤ 0.001) inhibited. Besides, the NK cells' activity and phagocytosis (macrophage) were increased significantly (p ≤ 0.001). Overall, the promising results of this study indicated the significant immunomodulatory effect of nanocurcumin-based formulation compared to the curcumin, which could be used against various inflammatory disorders such as allergy, asthma, autoimmune diseases, coeliac disease, inflammatory bowel disease, etc.

Effect of a Novel Ashwagandha-based Herbomineral Formulation on Pro-inflammatory Cytokines Expression in Mouse Splenocyte Cells: A Potential Immunomodulator.

BACKGROUND: Herbomineral formulations are momentous in an audience of worldwide by virtue of their holistic approach to life. These formulations are widely used as complementary therapies in immunocompromised patients including cancer. Still, there is the need of cost-effective and safe herbomineral-based formulation that can modulate immune response by the regulation of cytokines cascades. OBJECTIVE: Current study, we investigated immunomodulatory effect of TEBEH in LPS-induced cytokines expression levels in mouse splenocytes in vitro. MATERIALS AND METHODS: The most effective and safe concentrations of TEBEH were chosen by determining the cell viability of splenocytes using MTT assay. The pro-inflammatory cytokines such as TNF-α, IL-1ß, MIP-1α, and IFN-γ were measured in cell supernatants using ELISA. RESULTS: MTT data showed TEBEH formulation was found safe up to 10.53 µg/mL. At noncytotoxic concentrations (0.00001053-10.53 µg/mL), TEBEH significantly (P ≤ 0.001) inhibited the expressions of TNF-α, IL-1ß, and MIP-1α in mouse splenocytes as compared with vehicle control. CONCLUSION: In summary, TEBEH may indeed promote an anti-inflammatory environment by suppression of pro-inflammatory cytokines. These observations indicated that TEBEH has potential effects in downregulating the immune system and might be developed as a useful anti-inflammatory product for various inflammatory disorders. SUMMARY: The present study was undertaken to evaluate an immunomodulatory effect of the herbomineral formulation in LPS-induced mouse splenocytes with the measurement of cytokines expression such as TNF-α, IL-1ß, MIP-1α and IFN-γ. The results showed that the expression of TNF-α, IL-1ß, and MIP-1α was significantly down-regulated while, IFN-γ was significantly up-regulated in mouse splenocytes. It is hypothesized that modulation of the proinflammatory cytokines might occur via NF-κB pathway. Therefore, the herbomineral test formulation might act as an effective anti-inflammatory and immunomodulatory product, and this can be used as a complementary and alternative treatment for the prevention of various types of inflammatory and auto-immune disorders Abbreviations used: LPS: Lipopolysaccharide, IL: Interleukin; NF-κB: Nuclear factor kappa-B, TNF-α: Tumor necrosis factor alpha, MIP-1α: Macrophage inflammatory protein-1α, IFN-γ: Interferon, MTT: 3-(4,5-diamethyl-2-thiazolyl)-2, 5-diphenyl-2Htetrazolium), ELISA: Enzyme linked immune sorbent assay, ANOVA: Analysis of variance.

Protective effects of tetrahydrocurcumin (THC) on fibroblast and melanoma cell lines in vitro: it's implication for wound healing.

The aim of this study was to investigate the role of tetrahydrocurcumin (THC) against various skin health parameters using in vitro human foreskin fibroblast and melanoma cell lines (i.e. HFF-1 and B16-F10). The study was assessed using cell viability by MTT assay, identification of extracellular matrix component in HFF-1 cell line (i.e. collagen, elastin and hyaluronic acid), melanin synthesis in B16-F10 cells, cell viability against UVB-induced stress in HFF-1 cells, and in vitro wound healing by the scratch assay. THC was found to be safe and nontoxic up to the concentration of 10 µg/mL with improved level of collagen (37.90%), elastin (90.1%), and hyaluronic acid (74.19%) at 1 µg/mL. Besides, melanin was significantly inhibition by 78.5% at the lowest THC concentration of 0.1 µg/mL. UVB-protection rate was significantly improved by 61.2% and improved cell viability by THC in HFF-1 cells, which indicated protection from photoaging. In addition, THC showed significant wound healing activity (78.51%) and greater migration of fibroblast in HFF-1 cells at different time period. It can be concluded from the study that THC can protect the skin matrix with improved extracellular component synthesis and would healing via collagen synthesis in the skin, which improved the skin elasticity and tightness. Overall, it might be suggested that THC can be used as a safe skin whitening agent, wounds management, cosmetic applications, and treating various skin-related disorders.

Erythromycin-induced genotoxicity and hepatotoxicity in mice pups treated during prenatal and postnatal period.

The main aim of this study was to investigate the oxidative stress, genotoxicity and cytotoxicity of erythromycin (EMC) in pups of treated dams in gestation as well as the lactation period (LP). The two doses of EMC were compared using intraperitoneal (i.p.) route in two different periods, that is, in gestation period and in the LP. The rationale behind selection of i.p. route is because of the fact that EMC gets degrades in acidic pH of the stomach. The doses of EMC used were clinically equivalent dose (CED; EMC 14.2 mg/kg, i.p.) and a lower dose (EMC 10 mg/kg, i.p.) than CED. EMC toxicities in mice pups were evaluated using various parameters such as micronucleus (MN) test in peripheral blood and bone marrow, malondialdehyde (MDA) assay, glutathione (GSH reduced) assay and histopathological assessment in liver tissue. The CED of EMC led to a significant increase in MDA and decreased in GSH concentration in pups' liver tissue in both gestation and LPs and also to a significant increase in MN frequency in both peripheral blood and bone marrow cells of pups. There were no significant toxicities at a lower dose than CED. There were also some chronic findings with liver histopathological examination at CED. It is thus concluded that EMC accentuates the oxidative stress, cytotoxicity and DNA damage in pups of their postnatal life; hence, EMC should be avoided in the pregnancy and also in the LP.

Platelet-activating factor (PAF)-antagonists of natural origin.

Presently herbal medicines are being used by about 80% of the world population for primary health care as they stood the test of time for their safety, efficacy, cultural acceptability and lesser side effects. The discovery of platelet activating factor antagonists (PAF antagonists) during these decades are going on with different framework, but the researchers led their efficiency in studying in vitro test models. Since it is assumed that PAF play a central role in etiology of many diseases in humans such as asthma, neuronal damage, migraine, cardiac diseases, inflammatory, headache etc. Present days instinctively occurring PAF antagonist exists as a specific grade of therapeutic agents for the humans against these and different diseases either laid hold of immunological or non-immunological types. Ginkgolide, cedrol and many other natural PAF antagonists such as andrographolide, α-bulnesene, cinchonine, piperine, kadsurenone, different Piper species' natural products and marine origin plants extracts or even crude drugs having PAF antagonist properties are being used currently against different inflammatory pathologies. This review is an attempt to summarize the data on PAF and action of natural PAF antagonists on it, which were evaluated by in vivo and in vitro assays.

Furosemide-induced genotoxicity and cytotoxicity in the hepatocytes, but weak genotoxicity in the bone marrow cells of mice.

Furosemide (FS) is a potent loop diuretic widely used in the management of fluid retention associated with cardiac, renal, and hepatic failure as well as for the treatment of hypertension. FS is a well-characterized and known hepatotoxin in both human and animal test systems. In this study, an attempt has been made to investigate the in vivo genotoxicity of FS at the hepatotoxic equivalent doses using the chromosomal aberration and the comet assay in the bone marrow cells of mice as the endpoints of evaluation. The animals were treated with FS at the doses of 2.5, 5, 10, 20, 40, and 80 mg/kg/body weight (bw) intraperitoneal (ip) for both single (24 h) and repeated dose (seven consecutive days) toxicity studies. FS toxicity in the hepatocytes was evaluated using the parameters, such as, alanine-/aspartate-aminotransferase (ALT/AST), single cell gel electrophoresis (comet), tissue histology, DNA fragmentation, and TUNEL assay as the endpoints. The results clearly demonstrate that FS produced toxic responses in the hepatocytes as evident from increased ALT/AST level, DNA damage, TUNEL positive cells and increased DNA fragmentation in mice in vivo. However, it is interesting that in bone marrow cells, FS did not induced structural chromosomal abberations, but produced mild DNA strand breaks as observed by the comet assay. So it is considered as weak genotoxic toward the bone marrow cells when compared to the hepatocytes of mice.

The target-specific transporter and current status of diuretics as antihypertensive.

The currently available diuretics increase the urinary excretion of sodium chloride by selective inhibition of specific sodium transporters in the loop of Henle and distal nephron. In recent years, the molecular cloning of the diuretic-sensitive sodium transporters at distal convoluted tubule has improved our understanding of the cellular mechanisms of action of each class of diuretics. Diuretics are tools of considerable therapeutic importance. First, they effectively reduce blood pressure. Loop and thiazide diuretics are secreted from the proximal tubule via the organic anion transporter-1 and exert their diuretic action by binding to the Na(+)-K(+)-2Cl(-) co-transporter type 2 in the thick ascending limb and the Na(+)-Cl(-) co-transporter in the distal convoluted tubule, respectively. Recent studies in animal models suggest that abundance of these ion transporters is affected by long-term diuretic administration. The WHO/ISH guidelines point out that diuretics enhance the efficacy of antihypertensive drugs and will most often be a component of combination therapy.

Huge ameloblastoma of jaw-A case report.

Ameloblastoma is a tumor of odontogenic epithelium. It is a tumour of intermediate malignant potential which lies in the gray zone between benign and malignant neoplasm. A huge ameloblastoma revealing benign cytological features in FNAC is being reported.Ameloblastoma arises from odontogenic epithelium. This tumor can occur at any age. Though traditionally divided as solid and cystic, nearly all ameloblatomas show some cystic change. This tumor shows invasive property and a remarkable tendency of recurrence. The cases showing distant metastasis are recognized as malignant ameloblastoma. Ameloblastic carcinoma is a tumor with microscopic features of ameloblastoma that displays malignant features at cytological level.([2]) It usually has aggressive course. A case of large ameloblastoma with slow clinical course and benign cytological as well as histological features is being reported.