Development of nanofunctionalized oxovanadium(IV) complex and its anticancer, antidiabetic, DNA cleavage and cell imaging studies.

VO(IV) complex is little toxic and highly effective than vanadium salts. A vanadyl metal complex from 8-formyl-7-hydroxy-4-methyl coumarin derivative has been synthesized and functionalized with copper nanoparticles. The Spectrochemical studies such as UV, FTIR, 1NMR and ESR spectra were recorded to characterize the ligand(CUAP), Vanadyl complex[VO(CUAP)SO4] and nano Cu-VO(IV)complex efficiently. The structural studies of vanadyl complex confirmed that the ligand coordinate with metal through nitrogen atom of azomethine, carbonyl oxygen and phenolic oxygen. ESR spectrum of vanadyl complex revealed the covalent nature. XRD pattern of nano Cu-VO(IV) complex indicated the crystalline nature and the average particle size was 20.91 nm. SEM image of nano Cu-VO(IV) complex showed that the nano particles accumulated to form spherical shaped particles. The particle size obtained from Transmission Electron Microscopy of nano functionalized metal complex is âˆ¼ 20 nm. It is closely matched to the particle size calculated from XRD results. Fluorescence of vanadyl complex and nano Cu-VO(IV) complex exhibit the emission from 270 to 900 nm range with significant fluorescence at âˆ¼ 750 nm. The DNA cleavage of all the compounds was evaluated using Agarose gel electrophoresis technique and showed greater cleavage of vanadyl complex. The anticancer activity of compounds was carried out against two cancer cell lines viz Human Breast Cancer Cell line (MCF-7) and Human Leukemia Cancer Cell Line(K-562). Oxovanadium complex exhibited good anticancer activities than ligand and nano-functionalized complex. The antidiabetic activities of vanadyl and nano functionalized complexes were studied against α-Amylase and ß-Glucosidase inhibition assay. In this study vanadyl complex showed higher inhibition activity on α-Amylase compared with standard Acarbose. The bioimaging of nano-functionalized metal complex showed high fluorescent properties. The molecular docking study of ligand and vanadyl complex showed greater docking results with CDK2 receptor.

Nano zinc Oxide-Ruthenium (III) complex of novel coumarin derivative: Synthesis, Characterization, DNA Cleavage, anticancer and bioimaging activities.

The uniqueness of nanofunctionalized metal complexes would play a prominent role in medicinal chemistry, especially in cancer chemotherapy. Among the metal based chemotherapeutic drugs, ruthenium complexes exhibit different oxidation states, lower toxicity and remarkable antitumor activities than other anticancer drugs. In this investigation bioactive ruthenium(III) complex has been synthesized from novel coumarin derivative and it is functionalized with nanostructured zinc oxide (ZnO) under well defined condition. An octahedral geometry is proposed for the ruthenium complex based on analytical and spectrochemical characterization techniques (UV, FT-IR and NMR). The g|| and g⊥ values obtained from the ESR spectrum indicated high symmetry and octahedral field around ruthenium ion. The XRD patterns of all synthesized compounds explicit average particle sizes are nanometer range. The morphology and particle size of Nano ZnO-Ru(III) complex is also confirmed by using SEM and TEM analysis. The nanofunctionalized material exhibited enhanced fluorescence emissions at 674 nm and 681 nm. The evaluation of DNA cleavage study indicated compounds were effectively cleaved supercoiled pUC18 DNA by gel electrophoresis method. The examinations of in-vitro anticancer activities of compounds were studied against human breast (MCF-7) and leukemia (K-562) cancer cell lines. The ruthenium complex showed greater effect against MCF-7 with < 10 IC50 value. Nowadays, the cell imaging property of nanofunctionalized materials receive more attention. The fluorescence microscopic images of Nano ZnO-Ru(III) complex displayed higher luminescence at 100 µg/ mL against L6 rat skeletal muscle cell line.

Antimicrobial efficacy of nanosilver and chitosan against Streptococcus mutans, as an ingredient of toothpaste formulation: An in vitro study.

CONTEXT: Dental caries is the most prevalent oral infection affecting the humankind worldwide, and Streptococcus mutans is the major microrganism involved in its pathology. Mechanical plaque control in children is not performed efficiently and thus necessitates the inclusion of certain antimicrobial agents in the toothpaste. AIM: To analyze the antibacterial efficacy of nanosilver, chitosan, and fluoride as an ingredient in the dentifrices against S. mutans strains and comparing them with each other. SUBJECTS AND METHODS: Pure culture of S. mutans strain (MTCC 890) was obtained from the Institute of Microbial Technology, Chandigarh, India. Nanosilver (Group 1)- and chitosan (Group 2)-containing toothpastes were obtained from the respective dealers, and fluoride (Group 3) toothpaste was obtained from the local market. The antimicrobial activity of the three toothpastes was determined by modified agar well diffusion method. Saline was kept as the control. STATISTICAL ANALYSIS USED: Unpaired t-test was done for intergroup comparisons. RESULTS: Statistically significant differences between Groups 1 and 2, Groups 1 and 3, and Groups 2 and 3 were seen. CONCLUSIONS: Nanosilver-containing toothpaste has the highest antibacterial efficacy against S. mutans, followed by fluoride- and chitosan-containing toothpaste.

Role of Aquaporin 0 in lens biomechanics.

Maintenance of proper biomechanics of the eye lens is important for its structural integrity and for the process of accommodation to focus near and far objects. Several studies have shown that specialized cytoskeletal systems such as the beaded filament (BF) and spectrin-actin networks contribute to mammalian lens biomechanics; mutations or deletion in these proteins alters lens biomechanics. Aquaporin 0 (AQP0), which constitutes ∼45% of the total membrane proteins of lens fiber cells, has been shown to function as a water channel and a structural cell-to-cell adhesion (CTCA) protein. Our recent ex vivo study on AQP0 knockout (AQP0 KO) mouse lenses showed the CTCA function of AQP0 could be crucial for establishing the refractive index gradient. However, biomechanical studies on the role of AQP0 are lacking. The present investigation used wild type (WT), AQP5 KO (AQP5(-/-)), AQP0 KO (heterozygous KO: AQP0(+/-); homozygous KO: AQP0(-/-); all in C57BL/6J) and WT-FVB/N mouse lenses to learn more about the role of fiber cell AQPs in lens biomechanics. Electron microscopic images exhibited decreases in lens fiber cell compaction and increases in extracellular space due to deletion of even one allele of AQP0. Biomechanical assay revealed that loss of one or both alleles of AQP0 caused a significant reduction in the compressive load-bearing capacity of the lenses compared to WT lenses. Conversely, loss of AQP5 did not alter the lens load-bearing ability. Compressive load-bearing at the suture area of AQP0(+/-) lenses showed easy separation while WT lens suture remained intact. These data from KO mouse lenses in conjunction with previous studies on lens-specific BF proteins (CP49 and filensin) suggest that AQP0 and BF proteins could act co-operatively in establishing normal lens biomechanics. We hypothesize that AQP0, with its prolific expression at the fiber cell membrane, could provide anchorage for cytoskeletal structures like BFs and together they help to confer fiber cell shape, architecture and integrity. To our knowledge, this is the first report identifying the involvement of an aquaporin in lens biomechanics. Since accommodation is required in human lenses for proper focusing, alteration in the adhesion and/or water channel functions of AQP0 could contribute to presbyopia.

Intact and N- or C-terminal end truncated AQP0 function as open water channels and cell-to-cell adhesion proteins: end truncation could be a prelude for adjusting the refractive index of the lens to prevent spherical aberration.

BACKGROUND: Investigate the impact of natural N- or C-terminal post-translational truncations of lens mature fiber cell Aquaporin 0 (AQP0) on water permeability (Pw) and cell-to-cell adhesion (CTCA) functions. METHODS: The following deletions/truncations were created by site-directed mutagenesis (designations in parentheses): Amino acid residues (AA) 2-6 (AQP0-N-del-2-6), AA235-263 (AQP0-1-234), AA239-263 (AQP0-1-238), AA244-263 (AQP0-1-243), AA247-263 (AQP0-1-246), AA250-263 (AQP0-1-249) and AA260-263 (AQP0-1-259). Protein expression was studied using immunostaining, fluorescent tags and organelle-specific markers. Pw was tested by expressing the respective complementary ribonucleic acid (cRNA) in Xenopus oocytes and conducting osmotic swelling assay. CTCA was assessed by transfecting intact or mutant AQP0 into adhesion-deficient L-cells and performing cell aggregation and adhesion assays. RESULTS: AQP0-1-234 and AQP0-1-238 did not traffic to the plasma membrane. Trafficking of AQP0-N-del-2-6 and AQP0-1-243 was reduced causing decreased membrane Pw and CTCA. AQP0-1-246, AQP0-1-249 and AQP0-1-259 mutants trafficked properly and functioned normally. Pw and CTCA functions of the mutants were directly proportional to the respective amount of AQP0 expressed at the plasma membrane and remained comparable to those of intact AQP0 (AQP0-1-263). CONCLUSIONS: Post-translational truncation of N- or C-terminal end amino acids does not alter the basal water permeability of AQP0 or its adhesive functions. AQP0 may play a role in adjusting the refractive index to prevent spherical aberration in the constantly growing lens. GENERAL SIGNIFICANCE: Similar studies can be extended to other lens proteins which undergo post-translational truncations to find out how they assist the lens to maintain transparency and homeostasis for proper focusing of objects on to the retina.

Aquaporin 5 knockout mouse lens develops hyperglycemic cataract.

The scope of this investigation was to understand the role of aquaporin 5 (AQP5) for maintaining lens transparency and homeostasis. Studies were conducted using lenses of wild-type (WT) and AQP5 knockout (AQP5-KO) mice. Immunofluorescent staining verified AQP5 expression in WT lens sections and lack of expression in the knockout. In vivo and ex vivo, AQP5-KO lenses resembled WT lenses in morphology and transparency. Therefore, we subjected the lenses ex vivo under normal (5.6mM glucose) and hyperglycemic (55.6mM glucose) conditions to test for cataract formation. Twenty-four hours after incubation in hyperglycemic culture medium, AQP5-KO lenses showed mild opacification which was accelerated several fold at 48 h; in contrast, WT lenses remained clear even after 48 h of hyperglycemic treatment. AQP5-KO lenses displayed osmotic swelling due to increase in water content. Cellular contents began to leak into the culture medium after 48 h. We reason that water influx through glucose transporters and glucose cotransporters into the cells could mainly be responsible for creating hyperglycemic osmotic swelling; absence of AQP5 in fiber cells appears to cause lack of required water efflux, challenging cell volume regulation and adding to osmotic swelling. This study reveals that AQP5 could play a critical role in lens microcirculation for maintaining transparency and homeostasis, especially by providing protection under stressful conditions. To the best of our knowledge, this is the first report providing evidence that AQP5 facilitates maintenance of lens transparency and homeostasis by regulating osmotic swelling caused by glucose transporters and cotransporters under hyperglycemic stressful conditions.