Phloretin induces G2/M arrest and apoptosis by suppressing the ß-catenin signaling pathway in colorectal carcinoma cells.

Colorectal carcinoma (CRC) is the third most prevalent cancer, causing a significant mortality worldwide. Present available therapies are surgery, chemotherapy including radiotherapy, and these are known to be associated with heavy side effects. Therefore, nutritional intervention in the form of natural polyphenols has been well recognised to prevent CRC. Phloretin, a known dihydrochalcone is present in apple, pear and strawberry. This has been proven to induce apoptosis in cancer cells and also exhibited anti-inflammatory activity, thus can be explored as a potential anticancer nutraceutical agent. This study demonstrated phloretin's significant in vitro anticancer activity against CRC. Phloretin suppressed cell proliferation, colony forming ability and cellular migration in human colorectal cancer HCT-116 and SW-480 cells. Results also revealed that phloretin generated reactive oxygen species (ROS) which provoked depolarization of mitochondrial membrane potential (MMP) and further contributed to cytotoxicity in colon cancer cells. Phloretin also modulated the cell cycle regulators including cyclins and cyclin-dependent kinases (CDKs) and halted cell cycle at G2/M phase. Moreover, it also induced apoptosis by regulating the expression of Bax and BCl-2. The Wnt/ß-catenin signaling is inactivated by phloretin by targeting the downstream oncogenes namely CyclinD1, c-Myc and Survivin which are involved in the proliferation and apoptosis of colon cancer cells. In our study we showed that lithium chloride (LiCl) induced the expression of ß-catenin and its target genes and the co-treatment of phloretin circumvent its effect and downregulated the Wnt/ß-catenin signaling. In conclusion, our results strongly suggested that phloretin can be utilized as a nutraceutical anticancer agent for combating CRC.

Phloretin suppresses intestinal inflammation and maintained epithelial tight junction integrity by modulating cytokines secretion in in vitro model of gut inflammation.

Ulcerative colitis is a type of inflammatory bowel disease which in long run can lead to colorectal cancer (CRC). Chronic inflammation can be a key factor for the occurrence of CRC thus mitigating an inflammation can be a preventive strategy for the occurrence of CRC. In this study we have explored the anti-inflammatory potential of phloretin, in in vitro gut inflammation model, developed by co-culture of Caco2 (intestinal epithelial) cells and RAW264.7 macrophages (immune cells). Phloretin is a dihydrochalcone present in apple, pear and strawberries. An anti-inflammatory effect of phloretin in reducing LPS induced inflammation and maintenance of transepithelial electric resistance (TEER) in Caco2 cells was examined. Paracellular permeability assay was performed using Lucifer yellow dye to evaluate the effect of phloretin in inhibiting gut leakiness caused by inflammatory mediators secreted by activated macrophages. Phloretin attenuated LPS induced nitric oxide levels, oxidative stress, depolarization of mitochondrial membrane potential in Caco2 cells as evidenced by reduction in reactive oxygen species (ROS), and enhancement of MMP, and decrease in inflammatory cytokines IL8, TNFα, IL1ß and IL6. It exhibited anti-inflammatory activity by inhibiting the expression of NFκB, iNOS and Cox2. Phloretin maintained the epithelial integrity by regulating the expression of tight junction proteins ZO1, occludin, Claudin1 and JAM. Phloretin reduced LPS induced levels of Cox2 along with the reduction in Src expression which further regulated an expression of tight junction protein occludin. Phloretin in combination to sodium pyruvate exhibited potential anti-inflammatory activity via targeting NFkB signaling. Our findings paved a way to position phloretin as nutraceutical in preventing the occurrence of colitis and culmination of disease into colitis associated colorectal cancer.

Comparative Studies of Essential Oil Composition and Biological Activities of Callistemon citrinus from Western Himalaya.

This work described a comprehensive study to estimate chemical constituents of essential oils (EOs) extracted from different parts of the Callistemon citrinus viz: fruits, leaves and aerial part. The EOs were characterized using physicochemical parameters, and GC-FID/MS. It was observed that among different parts, aerial part has the highest oil yield (0.90 %) followed by leaves and fruits. Further, seventeen compounds were characterized, and represented total amount (97.2-99.5 %) with domination of monoterpenes (12.5-34.6 %) and oxygenated monoterpenes hydrocarbon (61.8-86.8 %). α-pinene (11.8-24.7 %), α-phellandrene (1.2-3.0 %), p-cymene (3.3-3.9 %) and 1,8-cineole (58.3-85.1 %) were found as major compounds in C. citrinus samples. These major compounds are the quality chemical markers of C. citrinus oil. The findings revealed significant quantitative variations in EO composition of samples and were also clearly supported by multivariate statistical analysis. Moreover, EOs were evaluated for glucosidase and colon cancer cell lines inhibitory activities, which were found promising.

Lysine and citric acid based pegylated polymeric dendritic nano drug delivery carrier and their bioactivity evaluation.

The main objective of this work is to synthesize multifunctional nanodendritic structural molecules that can effectively encapsulate hydrophilic as well as hydrophobic therapeutic agents. Four different types of fourth-generation lysine-citric acid based dendrimer have been synthesized in this work: PE-MC-Lys-CA-PEG, TMP-MC-Lys-CA-PEG, PE-MS-Lys-CA-PEG, and TMP-MS-Lys-CA-PEG. The antibacterial drug cefotaxime (CFTX) was further conjugated to these dendrimers. The dendrimer and drug-dendrimer conjugate structures were characterized with the help of FTIR,1H-NMR, and 13C-NMR spectroscopy. Zeta sizer, AFM, and HR-TEM techniques were used to investigate the particle size, surface topography, and structural characteristics of drug-dendrimer conjugates. In vitro drug release was then investigated using dialysis method. Various kinetic drug release models were examined to evaluate the type of kinetic drug release mechanism of the formulations. Cytotoxicity study revealed that the dendrimers encapsulated with CFTX exhibited 2-3% toxicity against healthy epithelial cells, indicating their safe use. Plain dendrimers show 10-15% hemolytic toxicity against red blood cells (RBC), and the toxicity was reduced to 2-3% when CFTX was conjugated to the same dendrimers. The 3rd and 4th generation synthesized drug-dendrimer conjugates exhibit a significantly effective zone of inhibition (ZOI) against both Gram-positive and Gram-negative bacteria. For Gram-positive bacteria, the lower concentration of 0.1 mg/mL showed more than 98% inhibition of drug-dendrimer conjugate samples against B. subtilis and more than 50% inhibition against S. aureus using 0.2 mg/mL, respectively. Moreover, samples with concentrations of 0.5 and 1.0 mg/mL exhibited more than 50% inhibition against S. typhimurium and E. coli, respectively.

Comprehensive polyphenolic profiling, antioxidant, antimicrobial, and cytotoxicity studies of Rhynchostylis retusa (L.) Blume: a medicinal orchid.

Rhynchostylis retusa (L.) Blume, commonly known as the Foxtail orchid, has garnered worldwide attention for its diverse medicinal properties. In this study, root extract and its fractions were evaluated for total polyphenols, flavonoids, targeted polyphenols, and antioxidant potential. The antimicrobial activity was assessed against Gram-positive and Gram-negative bacterial strains while cytotoxicity was assessed using the A549 and HCT-116 cell lines. The investigations showed that chloroform and ethyl acetate are the most effective solvents for fractionation of polyphenols from the parent extract. These fractions also exhibited strong antioxidant and cytotoxic potentials. The chloroform fraction showed maximum cell death of 87.35 and 92.36% in A549 and HCT- 116 cell lines respectively. All samples showed growth inhibition against bacterial strains except the n-hexane fraction, whereas the n-butanol fraction showed comparable antimicrobial activity with the tetracycline standard. The possible health benefits and thereby, application of R. retusa were thus revealed in this investigation.

Low-dose atropine 0.01% for the treatment of childhood myopia: a pan-India multicentric retrospective study.

OBJECTIVE: The objective of this study was to assess the efficacy of low-dose atropine 0.01% in controlling myopia progression among Indian children over a 2-year period. METHODS: This retrospective study, conducted across 20 centres in India, monitored the progression of myopia over 2 years after initiating treatment with 0.01% atropine eye drops. This included children between 6 and 14 years with baseline myopia ranging from -0.5 D to -6 D, astigmatism≤-1.5 D, anisometropia ≤ -1 D and documented myopia progression of ≥0.5 D in the year prior to starting atropine. Subjects with any other ocular pathologies were excluded. RESULTS: A total of 732 children were included in the data analysis. The mean age of the subjects was 9.3±2.7 years. The mean myopia progression at baseline (1 year before starting atropine) was -0.75±0.31 D. The rate of myopia progression was higher in younger subjects and those with higher baseline myopic error. After initiating atropine, myopia progression significantly decreased to -0.27±0.14 D at the end of the first year and -0.24±0.15 D at the end of the second year (p<0.001). Younger children (p<0.001) and higher baseline myopia (p<0.001) was associated with greater myopia progression and poor treatment response (p<0.001 for both). CONCLUSION: Low-dose atropine (0.01%) effectively reduces myopia progression over 2 years in Indian children.

Nuclear magnetic resonance-based metabolomics and cytotoxicity (HT-29 and HCT-116 cell lines) studies insight the potential of less utilized parts of Camellia sinensis (Kangra tea).

Camellia sinensis (tea) is an evergreen plant having bioactive compounds associated with various pharmacological effects, including anti-cancerous activity. These phytochemicals are variedly distributed in plant tissues. A detailed study to understand chemical composition within the economically underutilized tea tissues is required to generate value. Therefore, a comprehensive chemical profiling of underutilized C. sinensis parts [coarse leaves, flowers, fruits (immature);n = 9] was performed by NMR techniques. NMR (1D and 2D) spectroscopy ambiguously identified and quantified fifty-seven metabolites (Coarse leaves: 35, flowers; 42, immature fruits; 45). The statistical analysis showed apparent tissue-specific similarities (26 metabolites) and variations. Further, HPLC-DAD revealed absolute quantification of catechins, caffeine and theanine among the different parts of C. sinensis. Moreover, cytotoxicity studies of tea tissues against colorectal cancer cell lines showed anticancer potentials. This chemical information and anticancer activity of underutilized C. sinensis parts will help to develop value added nutraceutical and cosmeceutical products.

Seasonal variability in essential oil composition and biological activity of Rosmarinus officinalis L. accessions in the western Himalaya.

Rosmarinus officinalis L. is an imperative herb used in pharmaceutical yet knowledge on chemical and activity profile of essential oil (EO) to harvest seasons and accessions from the Himalayan region is limited. Thus, accessions were evaluated to determine the EO content, compositional, antimicrobial, and cytotoxic potential of rosemary in different harvest seasons during 2018‒2019. EO content was 30.5% higher in IHBT/RMAc-1 compared with IHBT/RMAc-2 accession while 27.9% and 41.6% higher in the autumn as compared with summer and rainy season, respectively. Major EO compound was 1,8-cineole; ranged from 32.50‒51.79% during harvest seasons and 38.70‒42.20% in accessions. EO was active against both the tested Gram-positive bacteria (Micrococcus luteus MTCC 2470 and Staphylococcus aureus MTCC 96). EOs showed inhibition of Gram-negative bacteria (Salmonella typhi MTCC 733), while Klebsiella pneumoniae MTCC 109 was found to be resistant. The rosemary EO of T1 (Rainy season IHBT/RMAc-1) was most effective against S. aureus MTCC 96 with the minimum inhibitory concentration (MIC) of 4% (v/v). In vitro cytotoxicity evaluation showed no potential anti-proliferative activity of EO. The rosemary EO profile in the western Himalayan region was influenced by harvesting seasons and genetic variability within the accessions; furthermore, a promising antibacterial agent in pharmaceutical and flavour industries.

Retrospective Study of Visual Outcomes and Complications After Sutureless, Flapless, and Glueless Intrascleral Fixation of Posterior Chamber Intraocular Lens in Children and Young Adults.

PURPOSE: To evaluate the visual outcomes and complications after sutureless, flapless, and glueless intrascleral fixation of a posterior chamber intraocular lens (IOL) in patients between the ages of 5 and 20 years. METHODS: Children with gross subluxated lenses and aphakia following intracapsular cataract extraction with inadequate posterior capsular support underwent intrascleral fixation of a standard three-piece posterior chamber IOL without sutures, flaps, or glue. A retrospective study of 15 eyes of 11 patients with a mean follow-up period of 6 months was performed. The main outcome measures were improvement in visual acuity (best corrected [BCVA] and uncorrected [UCVA]) and early and late postoperative complications. RESULTS: Preoperative to postoperative mean UCVA changed from 1.34 ± 0.35 to 0.36 ± 0.18 logarithm of the minimum angle of resolution (logMAR), respectively (P < .001). Preoperative to postoperative mean BCVA changed from 0.69 ± 0.37 to 0.19 ± 0.19 logMAR, respectively (P < .001). Two patients showed early postoperative complications within 1 week. One patient presented with vitreous hemorrhage and the other with hyphema on postoperative day 1. Both patients were treated conservatively and recovered by the first follow-up visit. None of the patients showed late postoperative complications during the 6-month follow-up period. The final visual outcome between the complication (n = 2) and no complication (n = 13) groups showed no statistically significant difference (P = .91). The final visual outcome between the eyes with primary and secondary implantation was similar with no statistically significant difference (P = .857). CONCLUSIONS: The technique of flapless, glueless, and sutureless scleral fixation of a three-piece IOL has shown significant improvement in visual acuity and does not cause significant complications postoperatively. [J Pediatr Ophthalmol Strabismus. 2017;54(6):381-386.].

Central retinal artery occlusion as a presenting manifestation of polycythaemia vera.

A 60-year-old woman with no premorbidities presented with symptoms of sudden painless vision loss in the left eye (LE). Best-corrected visual acuity in the LE was counting fingers close to face. A relative afferent pupillary defect was observed in the LE. Ocular fundus examination of LE was suggestive of central retinal artery occlusion. Systemic evaluation revealed splenomegaly and normal cardiac and carotid arteries. Haematological investigations revealed increased haemoglobin, haematocrit, platelet count and leucocytosis with low erythrocyte sedimentation rate (ESR). Features of myeloproliferative neoplasm were noted on bone marrow aspiration. An assay for JAK2 mutation was positive. Since erythropoietin levels were normal, a diagnosis of primary polycythaemia vera was made and treated with aspirin and phlebotomy twice weekly until the target haematocrit of under 45% was achieved.

Morphometric Analysis of Peripapillary Thickness in Hypertensives and Normotensives using Optical Coherence Tomography (OCT).

INTRODUCTION: Hypotension following antihypertensive treatment is associated with retinal nerve fibre loss. There are studies that have reported the changes in RNFL in hypertensives. AIM: To compare the Retinal Nerve Fibre Layer (RNFL) thickness using Optical Coherence Tomography (OCT) in hypertensive and normotensive individuals. MATERIALS AND METHODS: The cross-sectional study was conducted from October 2012 to September 2014 to compare the RNFL thickness using SD OCT in hypertensive {no other ocular or systemic co-morbidity, vision better or equal to 6/9 (n=30)} and normotensive (n=30) individuals. RESULTS: Statistically significant RNFL loss was detected in hypertensives (98.31 ± 7.01) when compared to the normotensives (102.51 ± 8.72) p=0.001. CONCLUSION: Significant loss of RNFL in hypertensives point towards the altered autoregulation and warrants larger studies to assess the clinical relevance.

Development and characterization of a new cell line CF from caudal fin of knifefish, Chitala chitala (Hamilton-Buchanan).

A new cell line was successfully obtained from caudal fin tissue of the economically important freshwater fish Chitala chitala. The cell line was optimally maintained at 28°C in Leibovitz's L-15 supplemented with 20% fetal bovine serum (FBS). The effects of temperature and concentration of FBS on the growth of CF cells were examined. The CF cell line consisted predominantly of fibroblastic-like cells. Moderately low plating efficiencies 8%, 11%, and 17% were observed, with CF cell line in L-15 Medium with 20% FBS. Chromosomal analysis of the cell line revealed a diploid number of 42 chromosomes in C. chitala. Molecular characterization of mitochondrial 16S rRNA and cytochrome oxidase subunit I confirmed the origin of the cell line. The cells were successfully cryopreserved in liquid nitrogen (-196°C) for 6 mo, and more than 85-90% of CF cells were revived.

Development and characterization of three new diploid cell lines from Labeo rohita (Ham.).

Development of cell lines from fish for identifying the pathogenesis of viral diseases and for vaccine production against viral and bacterial diseases is imperative where they are of commercial importance. Three new diploid fish cell lines (RF, RH, and RSB) were developed from fin, heart, and swim bladder of an Indian major carp, Labeo rohita, commonly called Rohu. All the cell lines were optimally maintained at 28 degrees C in Leibovitz-15 medium supplemented with 10% FBS. The propagation of RH and RSB cells was serum dependent, with a low plating efficiency (<16%), whereas RF cells showed 20% efficiency. The cytogenetic analysis revealed a diploid count of 50 chromosomes. The cells of RF and RSB were found to be epithelial, where as the cells of RH were mostly fibroblastic. The viability of the RF, RH, and RSB cell lines was 75, 70 and 72%, respectively after 6 months of storage in liquid nitrogen. The origin of the cell lines was confirmed by the amplification of 496 and 655 bp fragments of 16S rRNA and Cytochrome Oxidase Subunit I (COI) of mtDNA. The new cell lines would facilitate viral disease diagnosis and genomic studies.