Generation of Oxygen Free Radicals by Proflavine: Implication in Protein Degradation

Proflavine, an acridine dye, is a known DNA intercalating agent. In the present study, we show that proflavine alone on photoillumination can generate reactive oxygen species (ROS). These proflavine-derived ROS cause damage to proteins, and this effect is enhanced when the divalent metal ion Cu (II) is included in the reaction. Bathocuproine, a specific Cu (I) sequestering agent, when present in the reaction mixture containing Cu (II), was found to inhibit the protein degradation, showing that Cu (I) is an essential intermediate in the reaction. The effect of several scavengers of ROS such as superoxide dismutase, sodium azide, potassium iodide, and thiourea were examined on the protein damaging reaction. Potassium iodide was found to be the most effective in inhibiting protein damage followed by sodium azide and thiourea. Our results indicate the involvement of superoxide, singlet oxygen, triplet oxygen, and hydroxyl radicals in proflavine-induced damage to proteins

EXPLORATION OF PHYSICOCHEMICAL AND PHYTOCHEMICAL POTENTIAL OF MORINGA OLEIFERA LAM (SEHJANA) FRUITS/PODS

Background: The physicochemical evaluation is an important parameter in detecting adulteration or improper handling, therefore, physicochemical and phytochemical standardization is considered a prerequisite for the assessment of biological activity and determination of biological standards of the plant material, and it provides the analytical characteristics which may prove to be useful in fixing the physicochemical standard for herbal drugs. So, it becomes imperative to standardize the drugs to ensure their identity, quality and purity to ascertain therapeutic efficacy of herbal drugs. Objective: Therefore, the present study was aimed to evaluate the physicochemical and phytochemical standardization and quality control check of an important drug Sehjana (Moringa oleifera Lam) used for various diseases, Material and Methods: The test drugs, Sehjana (Moringa oleifera Lam.) were collected directly from the herbal garden of department of Ilmul Advia AMU, Aligarh. Which includes parameters recommended by National Unani Pharmacopeia Committee, Qualitative analysis and chromatographic studies (TLC) were performed for proper identification and quality control these parameter include Results: Ash values, Total ash, (6.44%) acid insoluble ash, (3.94%) water soluble ash, (2.53%) Successive extractive values in different solvent; petroleum ether (3.03%), diethyl ether (1.3%), chloroform (1.0%), acetone (1.5%), alcohol (13.1%), aqueous (18.56%), solubility in alcohol (15.2%) and water (22.93%), loss on drying (9.13%), pH at 1% (5.45), & 10% (4.81), bulk density (0.33%) and moisture content (8.6%). Conclusion: Preliminary phytochemical analysis of Sehjana (Moringa oleifera Lam.) showed presence of alkaloid, steroids, phenol, amino acid, and terpenoids, carbohydrate and protein, which may be active compound, responsible for its wide activities

Riboflavin Ameliorates Cisplatin Induced Toxicities under Photoillumination

BACKGROUND: Cisplatin is an effective anticancer drug that elicits many side effects mainly due to induction of oxidative and nitrosative stresses during prolonged chemotherapy. The severity of these side effects consequently restricts its clinical use under long term treatment. Riboflavin is an essential vitamin used in various metabolic redox reactions in the form of flavin adenine dinucleotide and flavin mononucleotide. Besides, it has excellent photosensitizing property that can be used to ameliorate these toxicities in mice under photodynamic therapy. METHODS AND FINDINGS: Riboflavin, cisplatin and their combinations were given to the separate groups of mice under photoilluminated condition under specific treatment regime. Their kidney and liver were excised for comet assay and histopathological studies. Furthermore, Fourier Transform Infrared Spectroscopy of riboflavin-cisplatin combination in vitro was also conducted to investigate any possible interaction between the two compounds. Their comet assay and histopathological examination revealed that riboflavin in combination with cisplatin was able to protect the tissues from cisplatin induced toxicities and damages. Moreover, Fourier Transform Infrared Spectroscopy analysis of the combination indicated a strong molecular interaction among their constituent groups that may be assigned for the protective effect of the combination in the treated animals. CONCLUSION: Inclusion of riboflavin diminishes cisplatin induced toxicities which may possibly make the cisplatin-riboflavin combination, an effective treatment strategy under chemoradiotherapy in pronouncing its antineoplastic activity and sensitivity towards the cancer cells as compared to cisplatin alone

On the emergence theme of physics

The book surveys mathematical relations between classical and quantum mechanics, gravity, time and thermodynamics from various points of view and many sources (with appropriate attribution). The emergence theme is developed with an emphasis on the meaning via mathematics. A background theme of Bohemian mechanics and connections to the quantum equivalence principle of Matone et al. is also developed in great detail. Some original work relating the quantum potential and Ricci flow is also included

A putative mechanism for calcitriol-Cu (II) interaction.

<p>(A) Production of hydroxl radicals, superoxide anions and hydrogen peroxide lead to DNA damage and consequent cell death. (B) Hetero-dimer of VDR (green) and RXR (orange) showing separation between the calcitriol binding site and bound DNA. Upper panel: View normal to axis of bound DNA. Lower panel: View along axis of bound DNA. Calcitriol (dark green sticks and spheres) bound to VDR, constitutes the free radical production site, is separated from the DNA molecule by ~39 Å. For reference, retinoic acid bound to RXR is also shown. The double-stranded DNA molecule is shown as thin black/blue sticks. Distance measurements and figure preparation were performed in PyMol (<u><a href="http://www.pymol.org" target="_blank">www.pymol.org</a></u>) using coordinates of the VDR/RXR hetero-dimer [Orlov et al. EMBO J. 2012] kindly provided by B. Klaholz.</p

Assay of DNA damage in COLs

<p>(A) DNA damage in control lymphocytes, vehicle control lymphocytes, COLs and COLs with Cu (II) (25µM). Values expressed as mean + SEM (n=3) ** P < 0.001as compared to control and vehicle control, P < 0.001 when * and ** are compared). (B) Removal of Cu (II) and DNA damage. COLs with Cu (II) (25µM) + neucuprione (Neo) (50µM) (membrane permeable Cu(II) chelator), desferroxamine (Desf) (50µM) (Fe(II) chelator), histidine (His) (50µM) (Zn(II) chelator) Values expressed as mean + SEM (n=3).*P < 0.001 when compared to control and COLs with Cu (II) (25µM). (C) COLs with Cu (II) (25µM) were incubated along with superoxide dismutase (SOD) (20µg/ml), catalase (Cat) (20µg/ml), and thiourea (Thio) (0.1mM) Values expressed as mean + SEM (n=3). (D) Representative images of Comet Assay (1) Control (2) Vehicle Control (3) COLs (4) COLs + Cu (II) (25µM) (5) COLs + Cu (II) (25µM) + Neo (50µM) (6) COLs+ Cu (II) (25µM) + Desf (50µM) (7) COLs+ Cu (II) (25µM) + His (50µM) (8) COLs+ Cu (II) (25µM) + SOD (20µg/ml) (9) COLs+ Cu (II) (25µM) + Cat (20µg/ml) (10) COLs+ Cu (II) (25µM) + Thio (0.1mM). *P < 0.001 when compared to control and COLs with Cu (II) (25µM). All incubations were carried out for 2 hours at 37º C.</p

Apoptosis: Number of Apoptotic cells (cells with nuclear fragmentation and/or with distinct apoptotic bodies) per hundred cells.

<p>(A) Control (B) Vehicle control (C) Cu (II) (25µM) [added to non calcitriol loaded, control lymphocytes] (D) Calcitriol overload (E) Calcitriol overload + Cu (II) (25µM) (F) Calcitriol overload + Cu (II) (25µM) + Bathocuprione (50µM) (membrane impermeable Cu(II) chelator) (G) Calcitriol overload + Cu (II) (25µM) + Neucuprione (50µM) (membrane permeable Cu(II) chelator) (H) Calcitriol overload + Cu (II) (25µM) + Desferoxamine (50µM) (Fe(II) chelator) (I) Calcitriol overload + Cu (II) (25µM) + Histidine (50µM) (Zn(II) chelator) (J) Calcitriol overload + Cu (II) (25µM) + SOD (20µg/ml) (K) Calcitriol overload + Cu (II) (25µM) + Catalase (20µg/ml) (L) Calcitriol overload + Cu (II) (25µM) + Thiourea (0.1mM). Values expressed as mean + SEM (n=3). P < 0.05 when compared to control and COLs with Cu (II) (25µM). All incubations were carried out for 2 hours at 37º C.</p

Histomicrographs of mice liver of major groups.

<p>Showing histomicrographs of liver samples of various groups indicated in parentheses. All the sections have been stained with Hematoxylin and Eosin stain and were snapped at 400×. (A) Control [group I] depicts normal hepatocellular structure and sinusoids (s). (B) Riboflavin treated [group II] shows features very akin to normal. (C) Cisplatin treated [group III] reveals microvesicular changes in the hepatocytes as well as regions devoid of hepatocytes (*). (D) The combination of cisplatin and riboflavin treated [group IV] again shows features quite similar to normal control.</p

AutoDock results (binding energy, inhibition constant and no. of hydrogen bonds) of ligand-L in B-DNA.

<p>AutoDock results (binding energy, inhibition constant and no. of hydrogen bonds) of ligand-L in B-DNA.</p