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The aim of this present study was to investigate the effect of chlorophyllin (CHL) on oxidative stress in Streptozotocine (STZ) induced diabetic mice. For the study, mice were divided into Group A: normal control, Group B: diabetic control, Group C: diabetic mice treated with the ascorbic acid, and Group D: diabetic mice treated with CHL. Levels of Reactive Oxygen Species (ROS), lipid peroxidation, protein carbonyl, superoxide dismutase (CuZn SOD &Mn-SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR) activities were examined in kidney and heart tissues of different experimental groups. Histological and ultrastructuralstudies were also carried out to evaluate any changes in tissues as well as sub-cellular organelles. ROS, lipid peroxidation, and protein carbonyl levels have been significantly decreased with concomitant increased of CuZn SOD, Mn-SOD, CAT, GPx, and GR activity in CHLtreated diabetic mice. The histological and ultrastructural studies showed that CHL attenuates the detrimental effect of oxidative stress and alleviated tissue injuries in STZ induced diabetic mice. These results suggested that CHL possesses antioxidative activity and has the potential to amelioratediabetes-associated oxidative stress in mice.

Protease inhibitor was isolated from the fruits of Solanum acculeatissimum Jacq. (SAPI) and purified by four steps. PI activity was assayed by estimating the residual hydrolytic activity of trypsin and chymotrypsin towards the substrates BAPNA (N-benzoyl-L-arginine-p-nitroanilide) and BTPNA (N-benzoyl-L-tyrosyl-p-nitroanilide), respectively, at pH 8.0 after pre-incubation with inhibitor. Molecular mass of purified PI was evaluated as 22.2 kDa by SDS–PAGE. Affinity column chromatography yielded a single prominent elution peak. The purity was further checked by RP-HPLC showing a single peak, coinciding with the protein peak. Specific activities of purified PI were 502 TlU and 433.7 CIU U/mg, towards the substrates of trypsin and chymotrypsin respectively with 0.95 mg protein content. Overall, the specific activities increased to 92.6 and 82.9 folds with 9.8 and 8.77 % yield with respect to the substrates. Subsequently, the larvicidal potential of different concentrations of purified SAPI was evaluated against the banana pest Odoiporus longicollis following the treatment on third, fourth and fifth instars larvae. The results of the total percent mortality (from 3rd to 5th instar and adult) demonstrated that the mortality rate progressively increased with the concentrations of SAPI (50 to 250 µM). A minimum of 8% mortality was observed with 50 µM concentration of the PI. It was significant to note that the percent deformity of adults noticed from 50 µM onwards was more than 2%. Thus, the present data suggests an eco-friendly approach for the control of agricultural pest, O. longicollis. Further studies are planned to trace the molecular mechanism of larvicidal potential of SAPI against the pest.

The antioxidant activity of methanolic (MTBL) and aqueous (ATBL) extracts of Terminalia bellerica leaves (TBL) was evaluated by various in vitro techniques. IC50 values of MTBL and ATBL were found to be 93 and 301 μg mL-1 for FRSA, 42 and 252 μg mL-1 for SARSA, 91 and 376 μg mL-1 for LPO, 53 and 540 μg mL-1 for HRSA, 142 and 544 μg mL-1 for FTC, respectively. RP was detected to be 2.72 and 10.49 ASE mL-1 for MTBL and ATBL. The MTBL showed better antioxidant activity than ATBL when compared with standard quercetin. TPC for MTBL and ATBL were 209.28 and 173.9 g kg−1 of GAE, respectively. In conclusion, methanolic extract of TB leaf has higher antioxidant activity than the aqueous extract and it could be a good source of natural antioxidant.

type:Journal Article, Electrons utilized in the heme oxygenase (HO) reaction are provided by NADPH-cytochrome P450 reductase (CPR). To investigate the electron transfer pathway from CPR to HO, we examined the reactions of heme and verdoheme, the second intermediate in the heme degradation, complexed with rat HO-1 (rHO-1) using a rat FMN-depleted CPR; the FMN-depleted CPR was prepared by dialyzing the CPR mutant, Y140A/Y178A, against 2 M KBr. Degradation of heme in complex with rHO-1 did not occur with FMN-depleted CPR, notwithstanding that the FMN-depleted CPR was able to associate with the heme-rHO-1 complex with a binding affinity comparable with that of the wild-type CPR. Thus, the first electron to reduce the ferric iron of heme complexed with rHO-1 must be transferred from FMN. In contrast, verdoheme was converted to the ferric biliverdin-iron chelate with FMN-depleted CPR, and this conversion was inhibited by ferricyanide, indicating that electrons are certainly required for conversion of verdoheme to a ferric biliverdin-iron chelate and that they can be supplied from the FMN-depleted CPR through a pathway not involving FMN, probably via FAD. This conclusion was supported by the observation that verdoheme dimethyl esters were accumulated in the reaction of the ferriprotoporphyrin IX dimethyl ester-rHO-1 complex with the wild-type CPR. Ferric biliverdin-iron chelate, generated with the FMN-depleted CPR, was converted to biliverdin by the addition of the wild-type CPR or desferrioxamine. Thus, the final electron for reducing ferric biliverdin-iron chelate to release ferrous iron and biliverdin is apparently provided by the FMN of CPR., source:http://www.jbc.org/content/281/42/31659.long

Metabolic syndrome, obesity and type 2 diabetes are metabolic disorders characterized by the insulin resistance and the impairment in the insulin secretion. Since impairment in the oxidative stress and adipocyte metabolism contribute to the formation of obesity and diabetes, targeting adipose tissue can be considered as an effective approach to fight against them. Rosiglitazone is used for treatment for patients with type 2 diabetes via inducing lipogenesis and transdifferentiation of white adipose tissue into brown adipose tissue. Since the development of such therapeutics is required to control the formation and function of brown fat cells, we aimed to reveal possible molecular mechanisms behind rosiglitazone induced biochemical changes in the adipose tissue. Cells were expanded in the adipocyte culture medium supplemented with 5 μg/mL insulin following 2 days' induction. After those cells were treated with rosiglitazone 0, 0.1 3 mol/L and 10 μmol/L rosiglitazone for 48 hours and at 8th day, cells were collected and stored at -80 °C. Then the cells were used to evaluate antioxidant enzyme activities, mineral and trace element levels and fatty acid composition. Glucose-6-phosphate dehydrogenase and glutathione reductase significantly reduced in rosiglitazone-treated groups compared to the control. Na, Mg, K, Ca, Cr, Fe, Ni, Cu, Zn, Rb, Sr, Cs, Ba and Pb were determined in the cell lysates via ICP-MS. Also, relative FAME content decreased in the rosiglitazone-treated groups compared to the control. Rosiglitazone treatment at low doses showed promising results which may promote brown adipose tissue formation.

Testis unique lipid environment is vital for fertility potential. Prenatal ethanol (EtOH) exposure impairs, but dietary docosahexaenoic acid (DHA) supports testis functions. However, these impacts on the developmental profile are less investigated. This study examined the effect of dietary DHA on testis function in relation to testis lipid composition in rats exposed to prenatal EtOH. Female Sprague-Dawley rats were exposed to EtOH (3g/kg twice a day) or dextrose throughout the gestational period, while dams and their pups were fed with a semi-purified diet supplemented with and without DHA (1.4%, w/w fatty acids) throughout the study. Samples were collected at gestational day (GD) 20, postnatal day (PD) 4, PD21, PD49, and PD90 for further analysis. As the testis developed, n-6 polyunsaturated fatty acids (PUFA), mainly docosapentaenoic acid (DPA) and n-6 very long chain fatty acids (VLCFA) increased markedly replacing n-3 PUFA, predominately DHA and MUFA. The triacylglycerides (TAG) fraction contained the highest proportion of both n-6 DPA and total n-6 VLCFA, increasing from PD21 to PD90. Dietary DHA increased the levels of fetal serum testosterone and sperm with normal morphology in adulthood, regardless of EtOH exposure. The testis DHA levels increased in DHA-treated groups, regardless of EtOH exposure and testis maturation, replacing n-6 DPA and total n-6 VLCFA only in immature testis up to PD21. Overall, the early provision of DHA in the diet can be a positive factor for male fertility by affecting sperm quality in adulthood, likely by influencing testis DHA content and testosterone levels in the fetal period. Marked increases in testicular n-6 DPA and n-6 VLCFA accumulation as the testis matures, mostly in TAG, indicates their potential role in testis development. The minimal impact of moderate levels of prenatal EtOH exposure on testis function might be related to an energy-dense diet used in this study that warrants further attention in future.

Collagen is the major structural scaffold in the body and serves as barrier between tissues, and thus its turnover is tightly regulated. Collagen triple-helical structure renders it resistant to general proteolysis. Several proteases are capable of cleaving the triplehelical regions of collagen, including several mammalian matrix metalloproteinases (MMPs) and bacterial collagenases. MMP-mediated collagenolysis is associated with numerous diseases and some bacterial collagenases have found clinical application use due to their efficiency in the hydrolysis of the collagen triple-helix. A selective Förster resonance energy transfer triple-helical peptide (fTHP) probe for monitoring the activity of Clostridial collagenase has been developed. The fTHP [sequence: Gly-mep-Flp-(Glyvi Pro-Hyp)4-Gly-Lys(Mca)-Thr-Gly-Pro-Leu-Gly-Pro-Pro-Gly-Lys(Dnp)-Ser-(Gly-Pro-Hyp)4-NH2] was stable at 37 °C and was efficiently hydrolyzed by bacterial collagenase (kcat/KM = 25,000 s -1 M-1) but not by clostripain, trypsin, neutral protease, thermolysin, or elastase. The bacterial collagenase fTHP assay can be utilized in applications where specific activity towards triple-helical collagen needs to be evaluated, such as isolation of cells from various tissues. An fTHP scaffold was also utilized to evaluate the sequence preferences of eight MMPs. Residues spanning from P3 to P11 investigated using a positional scanning synthetic combinatorial library. Deconvolution of the library data revealed distinct motifs for several MMPs and discrimination among closely related MMPs. The results of this study show that the P10 11 substrate play an important role in the collagenase-substrate interactions and that modifying these residues can drastically affect the affinity of MMPs towards THP substrates. The identified sequence preferences of MMPs will enable the design of selective triple-helical MMP probes that could be used for monitoring in vivo enzyme activity and enzyme-facilitated drug delivery., 2019, Includes bibliography., Degree granted: Dissertation (Ph.D.)--Florida Atlantic University, 2019., Collection: FAU Electronic Theses and Dissertations Collection

Objective: to investigate the diagnostic value of nesfatin-1 in cases of intestinal ischemia and ischemia/reperfusion. Study Design: An experimental study. Place and Duration of Study: the Experimental Animals Laboratory of Bezmialem University, in June 2018. Methodology: twenty-one healthy male Sprague Dawley rats were randomly divided into three groups of 7 rats each. In group 1: 1-hour intestinal ischemia followed by 5-hour reperfusion was performed. In group 2: rats were subjected to 6-hour intestinal ischemia. In group 3: rats underwent laparotomy and closure without performing any further procedure. Changes in leukocyte count, amylase, blood sugar, LDH, SGOT, CRP, and nesfatin-1 levels were determined. For histopathological examination, a small intestinal sample was taken and preserved in 10% formaldehyde. Results: nesfatin-1 value in group 2 was significantly higher than that in group 1 and group 3 (p=0.005, and p <0.001 respectively). Nesfatin-1 value in group 1 was significantly higher than that in group 3. A significant (r = 0.864/p <0.001) positive correlation was observed between nesfatin-1 value and pathology score. The pathology score of group 2 was significantly higher than that of group 1 and group 3 (p <0.001). Conclusion: serum nesfatin-1 can be a biomarker in acute mesenteric ischemia., NA

Butyl p-hydroxybenzoic acid, also known as butylparaben (BP), is one of the most common parabens absorbed by the skin and gastrointestinal tract and metabolised in the liver and kidney. Recent in vivo and in vitro studies have raised concern that BP causes reproductive, development, and teratogenic toxicity. However, BP-induced oxidative stress and its relation to tissue damage has not been widely investigated before. Therefore, we aimed to investigate the effects of butyl 4-hydroxybenzoate on enzyme activities related to the pentose phosphate pathway and on glutathione-dependent enzymes such as glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6-PGD), glutathione reductase (GR), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) in kidney, liver, brain, and testis tissues. Male rats were randomly divided into four groups to orally receive corn oil (control) or 200, 400, or 800 mg/kg/day of BP for 14 days. Then we measured G6PD, GR, GST, 6-PGD, and GPx enzyme activities in these tissues and studied histopathological changes. BP treatment caused imbalance in antioxidant enzyme activities and tissue damage in the liver, kidney, brain, and testis. These findings are the first to show the degenerative role of BP on the cellular level. The observed impairment of equivalent homeostasis and antioxidant defence points to oxidative stress as a mechanism behind tissue damage caused by BP., Scientific Research Projects Coordination Unit of Hacettepe University

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Di (2-ethylhexyl) phthalate (DEHP) is used as plasticizer in the industry and belongs to the phthalate family which can induce tissue damage including kidney, liver, and testis as a result of elevated oxidative stress levels. Glutathione reductase (GR), Glucose-6-phosphate dehydrogenase (G6PD), glutathione S-transferase (GST), 6-phosphogluconate dehydrogenase (6PGD), enzyme activities, trace element and mineral levels were evaluated in the brain and testis tissue samples. Our data revealed that, antioxidant enzyme activities in the brain and testis samples were statistically insignificant in the DEHP administered groups compared to the control group except 400 mg/kg/day DEHP dose group in the testis samples. DEHP can disrupt trace element and mineral levels unlike antioxidant enzyme levels that may due to blood-brain and testis-blood barrier and/or short-term exposure to the DEHP. For more detailed information than the data presented in this Journal article, please see the research Journal article “Impact of the di (2-Ethylhexyl) phthalate administration on trace element and mineral levels in relation of kidney and liver damage in rats” [1]., Scientific Research Projects Coordination Unit of Hacettepe University

Background: the study aimed to investigate whether timolol-treatment has a beneficial effect on pentose phosphate pathway enzyme activities such as glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGDH) enzyme activities and cAMP level in streptozotocin-induced diabetic rats in pancreatic tissues. Methods: diabetes was induced by streptozotocin (STZ) in 3-month old male Wistar rats. The diabetic rats were treated with timolol (5 mg/kg body weight, for 12 weeks) while the control group received saline. Enzyme activities were determined in pancreas tissue. To support our results, we performed in silico calculations, using Protein Data Bank structures. Results: timolol treatment of STZ-induced diabetic rats had no noteworthy effect on high blood-glucose levels. However, this treatment induced activities of G6PD and 6PGDH in diabetic rats. Timolol treatment significantly increased cAMP level in diabetic pancreatic tissue. We found that timolol cannot bind strongly to either G6PD or 6PGD, but there is a relatively higher binding affinity to adenylyl cyclase, responsible for cAMP production, serving as a regulatory signal via specific cAMP-binding proteins. Conclusions: our data point out that timolol treatment has beneficial effects on the antioxidant defence mechanism enzymes in the pancreas of STZ-induced diabetic rats. / Uvod: cilj istrazivanja je bio da se utvrdi da li tretman timololom ima pozitivan efekat na aktivnosti enzima pentoze fosfata, kao sto su aktivnosti glukoze-6-fosfat dehidrogenaze (G6PD), enzimske aktivnosti 6-fosfoglukonat dehidrogenaze i cAMP nivo u tkivu pankreasa kod pacova kojima je dijabetes izazvan streptozotocionom. Metode: dijabetes je izazvan streptozotocionom (STZ) kod tromesecnih muzjaka vistar pacova. Pacovi sa dijabetesom su tretirani timololom (5 mg/kg telesne tezine tokom 12 nedelja), dok je kontrolna grupa primila fizioloski rastvor. Enzimske aktivnosti su utvrivane u tkivu pankreasa. Da bismo potkrepili na, Research Infrastructure Support Program of the Ministry of Development of the Republic of Turkey

Introduction: Diabetes mellitus (DM) is a group of metabolic diseases characterized by hyperglycemia resulting from defects in insulin secretion, insulin action, or both. The chronic hyperglycemia is associated with long-term damage, dysfunction, and failure of various organs, especially the eyes, kidneys, nerves, heart, and blood vessels and is among the top ten causes of death in the world. Borassus aethiopum Mart. (family Arecaceae) is a plant species of Borassus palm found widely across Africa. It serves an important source of food, providing edible fruits, and nuts, and also has a number of pharmacological uses that have been reported in some parts of the world. Objective: The current study was aimed at assessing the phytochemical constituents as well as the antidiabetic and hypolipidaemic effect of fruit extract of B. aethiopum in alloxan-induced diabetic rats.Materials and Methods: The fruits extract was prepared (FEB) and phytochemical constituents evaluated using standard methods. The antidiabetic and hypolipidaemic properties in alloxanized rats assessed for 7 and 28 days. Normoglycaemic and alloxan-induced diabetic rats were treated with FEB at doses of 100 mg, 250 mg and 500 mg/kg body weight. Observations on body weight, relative organ weight, haematological and biochemical parameters were measured in both acute and sub-chronic circumstances. Results: The presence of tannins, saponins, glycosides, triterpenoids and alkaloids were detected. Fasting blood glucose was reduced significantly (p<0.05) in diabetic rats in acute study at a dose of 500 mg/kg body weight and at 250 mg and 500 mg/kg body weight in sub-chronic studies. WBC and PLT levels were significantly increased after treatment with 500 mg/kg body weight. Urea and ALT levels also reduced significantly in both acute and sub-chronic studies. Conclusion: The aqueous fruit extract of B. aethiopum is antidiabetic. It was also found to be nephron- and hepato-protective as well as boosting the immu

Cancer is a difficult disease to treat, and few effective drugs are available. Hence, it is of great importance to develop effective anti-cancer therapeutic agents with well-defined pharmacokinetic properties. Although, ginger (Zingiber officinale) has a number of proven pharmacological activities, its effect on colon cancer has not received much attention. This study therefore investigated the potential colon cancer drug of compounds found in ginger. Dihydropyrimidine dehydrogenase was modeled using comparative homology modeling and virtual screening was performed locally on a Linux platform using AutoDock Vina®. The results showed that human dihydropyrimidine dehydrogenase is a homolog of pig dihydropyrimidine dehydrogenase. The leads of potential colon cancer drugs were beta-sitosterol, 6-Shogoal, Alloaromadedrene, and Zingiberol. They had similar binding site with levamisole for tumor necrosis factor ligand superfamily member 6 with His 148 and Tyr 192 common at their binding site whereas they had different binding sites from 5-fluorouracil for dihydropyrimidine dehydrogenase. The leads had better bioactivities compared with reference drugs (5-flourouracil and Levamisole) approved clinically for the treatment of colon cancer. In vitro, ex vivo and/or in vivo validations of the leads against colon cancer are recommended.

Breast cancer is the most common cancer and a major cause of death in women. The present study was designed to evaluate the antioxidant and anticancer potential of cranberry extract against N-methyl-N-nitrosourea (MNU) induced mammary carcinoma in rats. The tumor was induced in Female virgin rats of age 50 days by single dose of MNU (50mg/kg.b.w i.p.). After 85 days; all rats developed at least one tumor. Animals were treated with cranberry extract (400 and 600 mg/kg.b.w.orally) and tamoxifen (2mg/kg.b.w. i.p) for 4 weeks (from day 86 to day 113). MNU treatment resulted in a significant decrease (p < 0.05) in blood hemoglobin (Hb), red blood cells (RBC), platelets (PLTs) as well as blood, liver and breast catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD). However, MNU treatment resulted in a significant increase in White blood cells (WBC) as well as plasma, liver and mammary tissue gamma glutamyl transferase (GGT), lactate dehydrogenase (LDH), hexosamine, sialic acid and thiobarbituric acid reactive substances (TBARs). Upon administration of the cranberry extract, the levels of WBC, GGT, LDH, hexosamine, sialic acid, TBARs, Hb, RBC, PLTs, CAT, GPx and SOD were significantly normalized. Histopathological changes also confirmed the formation of tumor tubules and neovascularization after the MNU treatment. Cranberry extract administration significantly reduces the growth of MNU-induced mammary tumors, and therefore has strong potential as a useful therapeutic regimen for inhibiting breast cancer development. Comparing the beneficial effect of cranberry extract with that of MNU-induced breast cancer, cranberry extract showed antitumor and antioxidant activity indicated by the measured biochemical parameters and the histopathological examination of mammary tissue. The results of the present study indicate that cranberry extract possesses strong anticancer effects through its role in modulating glycoprotein components and the levels of oxidat

Chemotherapy resistance in human epithelial ovarian cancer (EOC) is a significant reason for the high rate of death among patients. We hypothesized that chemotherapy- resistant EOC cells will be killed by novel drug treatments in non-adherent culture conditions. The objective of this study was to test the efficacy of novel drugs to affect platinum resistant EOC cell viability. To achieve this, the cell killing efficacy of several drugs were tested on drug-resistant EOCs growing in non-adherent cultures. Both EOC cell lines and primary EOC cells isolated from patient ascites were used for these studies. Two different classes of drugs were tested including multikinase inhibitors (dorsomorphin and LDN-193189), and an understudied class of novel chemotherapeutic agents called glycosylated antitumor ether lipids (GAELs). EOC cells were treated with the drugs at different doses alone or in combination with cisplatin. Because GAELs exhibited promising results in resistant EOC cells, the mechanism of GAEL-induced cell-death was evaluated.

Acridone is a unique naturally occurring alkaloid known to associate with several biological activities. 2,3-dimethoxy-10-methyl-10,8a-dihydroacridin-9(8aH)-one (4) and its precursor 2-((3,4-dimethoxyphenyl)methylamino)benzoic acid (3) were synthesized and investigated for potential antioxidant and inhibitory activity against acetylcholinestrase. The synthetic pathway involves reaction of 2-(methylamino) benzoic acid (1) with 4-chloro-1,2-dimethoxybenzene (2) in presence of CuO and K2CO3 to give the precursor 3. Subsequent, cyclcondensation of 3 with Conc. H2SO4 afforded the anticipated acridone 4. Furthermore, the dimethoxyacridone derivative 4 showed potent antiacetylcholinesterase (ACHE) activity at (100 uM) with IC50 = 9.25 uM that is as potent as the reference drug rivastigmine. Assessment of total antioxidant activity of compounds 3 & 4 in comparison to known standard compounds revealed the following order: α-tocopherol > Acridone 4 > trolox > butylated hydroxyl anisole (BHA) > butylated hydroxyl toluene (BHT) > compound 3. Molecular docking characteristics of 3 & 4 within the active site of AChE (PDB: 1ACJ) co-crystallized with 9-amino-tetrahydroacridine (Tacrine) have been studied. Interestingly, the results revealed comparable binding poses to the co-crystallized ligand and demonstrates good correlation of the binding energy (DG) with the observed IC50-values. This finding suggests that compounds 3 & 4 exhibit good antioxidant effect and inhibition of acetylcholinesterase, which might provide profitable candidates in management of Alzheimer’s disease.

Introduction: Indonesian male population has traditionally used Pimpinella alpina Molk (PaM) to prevent degenerative disease. However, the scientific evidence of PaM effect on increase in antioxidant and decrease in prooxidant level and their negative correlation remains unclear. Objective: To prove the effect of PaM on increase in antioxidant and decrease in prooxidant and their negative correlation following Ultraviolet B (UVB) exposed repeatedly. Methods: Forty male rats were assigned into 8 groups, treatment groups for 7 days: PaM 50 mg (PaM50-7), PaM 100 mg (PaM100-7), PaM 150 mg (PaM150-7), and for 15 days: PaM 100 mg (PaM100-15), PaM 150 mg (PaM150-15). The increase in antoxidant and decrease in prooxidant levels were measured by ELISA and Spectophotometer. Results: Statistical analysis indicated that antioxidant Catalase (CAT) and Super Oxyde Dismutase (SOD) activities in PaM groups were significantly higher, p < 0.001. In contrary, prooxidant levels marked by Malondialdehide (MDA) and 8-hydroxy-2-deoxyguanosine (8OHdG) concentrations in PaM groups were significantly lower, p < 0.001. There was also a negative correlation between antioxidant and prooxidant levels, p < 0.001. Conclusion: PaM administration with 50-150 mg daily dosage for 7-15 days capable of increasing antioxidant and decreasing Prooxidant levels, with a negative correlation following UVB irradiation repeatedly.

Introduction: Incidence of breast cancer is high and constitutes a significant cause of female mortality. One of breast cancer type is Estrogen and Progesteron Receptor (ER and PR) positive, therefore affected by estrogen and progesteron concentration. Treatment of Thyponium flageliforme in combination with Curcuma zedoaria, and Phyllanthus niruri (TCP) have been proven capable of inhibiting tumor cells proliferation. Objective: To evaluate the expression of protein estrogen receptor and caspase-9 in C3H mice were inoculated with adenocarcinoma cells. Methods: 18 mice were inoculated with one million cells of breast adenocarcinoma per mouse and assigned into three groups. TCP with dosage 85 and 125 mg were treated for 21 days to evaluate the expression of protein estrogen receptor and caspase-9 measured by immuno-histochemical staining. Results: Mann Whitney analysis demonstrated that expression of protein estrogen receptor and caspase-9 in TCP 125 was significant (p < 0.05). Conclusion: Expression of protein estrogen receptor was decreased and caspase-9 expression was increased by TCP administration.

Introduction: People worldwide drink Zamzam water either medicinally or religiously. However, the effect of zamzam water extract of CZ in breast cancer cells remains unclear. Objective: To evaluate the zamzam water extract of CZ effect on cell apoptosis, p53 and Bcl2 protein expression in T47 D cells line culture. Methods: In the control group design, 16 Wells which was filled with T47 D cell line culture in the density of 5 × 104/100 μl, were assigned into 4 groups. Control group (C-G), were only filled with T47D cell line culture; Zamzam 50 (ZZ-50); Zamzam 100 (ZZ-100), and Zamzam 500 (ZZ-500) groups, were filled with T47D cell line cultures and 50, 100, and 500 μg/ml zamzam water extract of CZ respectively, and then incubated for 48 hours. The number of apoptotic cell was counted by flow cytometry, while p53 and Bcl2 measured by immunohistochemistry staining method. Results: Mann Whitney analysis indicated that the percentage of apoptotic cell and p53 protein expression in ZZ-100 was significant higher compared to that of C-G (p < 0.05). In contrary the percentage number of Bcl2 protein expression in ZZ100-G was significantly lower when compared to that of C-G (p < 0.05). Conclusion: Treatment of zamzam water extract of CZ on T47D cell line cultured was capable of increasing the number of apoptotic cells, p53 protein expression, but decreased Bcl2 protein expression in T47D cell line culture. However the numbers of apoptotic cells were less than 10%.

This research is part of a long-term project aimed at elucidating important structural features, of both ligands and metals, that are needed to produce effective anti-cancer agents. The specific goal is the synthesis of organotin polymers containing amino acids, in this case the diamino acid diglycine. The desired materials were synthesized with percent yields ranging from 32-99%. The products were synthesized employing the interfacial polymerization technique. The polymers were then characterized utilizing the following physical characterization techniques: light scattering photometry (LS), Infrared spectroscopy (IR), nuclear magnetic resonance spectroscopy (NMR), and matrix assisted laser desorption mass spectroscopy (MALDI). Physical characterization showed evidence of formation of desired adducts in addition to data that was consistent with the formation of materials containing multiple repeat units. The materials were then analyzed for biological activity. The synthesized materials displayed the ability to inhibit tested cancer cell lines., 2018, Includes bibliography., Degree granted: Thesis (M.S.)--Florida Atlantic University, 2018., Collection: FAU Electronic Theses and Dissertations Collection

Raman spectroscopy based activities were developed and implemented into the first year chemistry undergraduate curriculum. The implementation of these experiences and experiments and the utility of Raman spectroscopy as a teaching tool to convey anchoring chemistry concepts using a hands-on Raman spectroscopy based approach are discussed. Fundamental principles of chemistry, such as the interaction of light with matter, molecular bonding, equilibrium, and acid base reactions are facilitated through use of these Raman spectroscopy based experiments and experiences. An assessment of student learning gains as a result of participation in a Raman spectroscopy experience was also conducted and is discussed., 2018, Includes bibliography., Degree granted: Thesis (M.S.)--Florida Atlantic University, 2018., Collection: FAU Electronic Theses and Dissertations Collection

This project was designed to investigate the effects of carbon dioxide (CO2) levels on the growth and pigment ratios (chemotaxonomy) of freshwater algal species typical to the south Florida surface waters. Green algae, diatoms, and cyanobacteria were cultured under 400 or 800 ppm CO3 in air for several weeks. Growth monitoring used a cell counter, hemocytometer, and chlorophyll fluorescence. Pigments were analyzed using HPLC-PDA. Experiments with certified CO2 concentrations (400, 600, 800, 1200 ppm) were conducted with helium degassed ultrapure water and each of three culture media. Theoretical and experimental pH values with water matched exactly. However, each culture media proved to exhibit significant buffer capacity. Cell growth monitoring was problematic except for the cyanobacterium Microcystis aeruginosa. That species responded to increased CO2 (800 ppm) with increased growth rates as predicted. The other species gave erratic results mainly due to difficulties in obtaining valid consistent cell counts., 2018, Includes bibliography., Degree granted: Thesis (M.S.)--Florida Atlantic University, 2018., Collection: FAU Electronic Theses and Dissertations Collection

The use of peptide drugs has gained popularity recently. Peptides are attractive drug targets due to their high specificity and potency towards their biological targets. A drawback for peptide drugs is a lack of stability for oral delivery. Two classes of disulfide-rich peptides, conotoxins and cyclotides, have been shown to have higher stability than linear peptides thanks to their disulfide connectivity. Conotoxins are present in the venom of cone snails, a carnivorous marine mollusk that preys upon fish, worms, or other mollusks. Conotoxins are promising drugs leads with great prospects in the treatment of diseases and disorders such as chronic pain, multiple sclerosis and Parkinson’s and Alzheimer’s diseases. Cyclotides, which are cyclic cysteine knot containing peptides, isolated from the Violaceae (violet), Rubiaceae (coffee), and Cucurbitaceae (cucurbit) families and they have a wide range of biological activities, such as anti-HIV, uterotonic, and antimicrobial. P-superfamily framework IX conotoxins (C-C- C-CXC- C) contain the same cysteine framework, homologous sequences, and similar 3D structures to cyclotides. The knot containing conotoxins have been identified in several Conus species, but this work focuses on those from Conus brunneus, Conus purpurascens, and Conus gloriamaris. The cysteine knot motif of cyclotides and P-superfamily conotoxins is characterized by a cyclic backbone and six-conserved cysteine residues that form the three-disulfide bridges of the “knot”. This motif provides cyclotides and conotoxins with superior stability against thermal, chemical, and enzymatic degradation; marking them as potential frameworks for peptide drug delivery. Presented are details on the isolation of conotoxins and cyclotides, from Viola tricolor, and the characterization of their activity in the well-characterized Drosophila melanogaster giant fiber system (GFS) neuronal circuit, which contains GAP, acetylcholine, and glutamate synapses. The transcriptomes of, 2018, Includes bibliography., Degree granted: Dissertation (Ph.D.)--Florida Atlantic University, 2018., Collection: FAU Electronic Theses and Dissertations Collection

Objective: This study investigated urinary nerve growth factor (NGF) and fractalkine levels in women with overactive bladder (OAB), as well as diagnostic and/or prognostic roles, and correlation of these urinary biomarkers with symptom severity of patients.Materials and Methods: Twenty-seven women with OAB and 26 healthy subjects were enrolled. Patients were diagnosed with OAB based on symptoms, a 3-day voiding diary and a validated Turkish version of the Overactive Bladder-Validated 8 (OAB-V8) questionnaire. The urinary baseline levels of NGF and fractalkine were compared between OAB patients and control group. The Turkish-validated International Consultation on Incontinence Questionnaire Short Form and OAB-V8 used to categorize patients according to disease severity to assess treatment efficacy. Further urinary NGF and fractalkine levels were compared before and after antimuscarinic treatment.Results: Urinary NGF/creatinine (Cr) and fractalkine/Cr were significantly elevated in OAB patients (0.40 ± 0.40 ng/mg and 4.63 ± 4.36 ng/mg, respectively) compared to healthy subjects (0.14 ± 0.08 ng/mg and 2.00 ± 1.29 ng/mg, P = 0.002 and P = 0.005, respectively). Sensitivity and specificity were 85.2%, 65.4% for NGF, and 74.1%, 65.4% for fractalkine, respectively. No significant differences in NGF/Cr and fractalkine/Cr compared to baseline (P = 0.063 and 0.162, respectively) were observed after trospium chloride treatment in OAB patients. NGF/Cr and fractalkine/Cr exhibited no correlations with symptom severity levels.Conclusions: Increased urinary NGF/Cr and fractalkine/Cr levels were found in OAB women. However, the sensitivity and specificity were not sufficient for diagnostic use. NGF and fractalkine levels were decreased after treatment insignificantly, and there was no correlation with symptom severity; therefore their prognostic worth was limited.

“Agnijith” a herbal preparation was evaluated against fifteen different microorganisms at different concentrations 120mg, 130mg, 150mg, 170mg and 190mg/100mg. Chloromphenicol was used as control. The in vitro antibacterial activity was performed by agar well diffusion method and minimum inhibitory concentration (MIC) method. “Agnijith” was found to be more potent in inhibiting the microorganisms such as Bacillus subtilis, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Pseudomonas flourescence, Proteus mirabilis and Shigella dysentriae, whereas no inhibition zone was found against Micrococcus luteus, Streptococcus pyrogens, Enterococcus faecalis, Proteus vulgaris and Salmonella typhimurium by well diffusion method. The zone of inhibition ranges from 12mm to 25mm. Bacillus subtilis, Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Pseudomonas flourescence, Shigella dysentriae were found to be sensitive at various concentrations by minimum inhibitory concentration method. The results clearly demonstrates that the “Agnijith” to be an effective antibacterial agent and could be used to treat diseases caused by microorganisms.

The research of the effects of different doses of ascorbic acid (AA) on activity of lactate dehydrogenase (LDG) in the organs of white nonlinear rats with hypoxia of closed space (HCS) has been conducted. Injections of АА to rats for 30 minutes of HCS reduced the LDG activity in all organs of hypoxic animals, Проведено дослідження дії різних доз аскорбінової кислоти (АК) на активність лактатдегідрогенази (ЛДГ) в органах білих нелінійних щурів при гіпоксії замкненого простору (ГЗП). АК, яку вводили щурам за 30 хв. до створення ГЗП, різним чином зменшувала активність ЛДГ у всіх органах у порівнянні з показниками гіпоксичних тварин.

Enicostemma littorale leaves are traditionally used for the treatment of several diseases, including inflammation and cancer. This study has taken effort to explore the antitumor initiating potential of E. littorale leaves (ElELet) by analyzing the expression pattern of apoptotic (p53, Bcl-2 and Bcl-2 associated X-protein), cell-proliferative (cyclin D1 and proliferating cell nuclear antigen), angiogenic (vascular endothelial growth factor), invasive (matrix metalloproteinase-2 and 9), and inflammatory (NF-κB and cyclooxygenase-2) markers during 7, 12-dimethylbenz (a) anthracene (DMBA) induced hamster buccal pouch carcinogenesis. Oral tumors were induced in the buccal pouches of hamsters using the potent site and organ specific carcinogen, DMBA. DMBA application 3 times a week for 14 weeks resulted in tumor formation in the buccal pouches. Hundred percent tumor formations with dysregulation in the expression pattern of apoptotic, cell proliferative, inflammatory, angiogenic, and invasive markers were observed in the buccal pouches of hamsters treated with DMBA alone. ElELet at a dose of 250 mg/kg body weight orally to DMBA treated hamsters significantly prevented the tumor formation as well as corrected the abnormalities in the expression pattern of above mentioned molecular markers. ElELet thus modulated the expression pattern of all the above mentioned molecular markers in favor of the suppression of cell proliferation occurring in DMBA induced hamster buccal pouch carcinogenesis.

Background: Pergularia daemia (Asclepiadacea) is a fetid- smelling perennial herb growing well along the river bang and road sides of India. Naturally the plant has powerful antioxidants including polyphenols, flavanoids, steroids and terpenoids. Objective: The aim of this study is to evaluate the in vitro antioxidant potential and to determine the median lethal dose (LD50) of crude ethyl acetate and methanol extracts of Pergularia daemia. The plant Pergularia daemia possess effective scavenging activity against 2, 2’ azino bis (3 ethylbenzothiazoline 6 sulfonic acid (ABTS), nitric oxide and reducing power radicals at different concentrations (100, 200, 300, 400 & 500 μg/mL) of both extracts. Results: From the above in vitro assay we have exposed that the methanolic extract exert higher antioxidant activity at 400 μg/mL than ethyl acetate extract. Acute toxicity study revealed that the extracts showed no signs of toxicity upto a dose level of 2500 mg/kg b.wt. Conclusion: Thus our findings provide that both extracts of Pergularia daemia possess a strong antioxidant capacity and are relatively has high margin of safety.

Calcium Adenosine Triphosphatase is a ubiquitous enzyme and key protein for the maintenance of calcium homeostasis in various cellular compartments. Moringa leave has been reported to be a very good source of dietary protein as well as pool of minerals. So far the effect of varying dietary protein on the activities of Ca2+ pump across cell membranes is yet to be fully explained. In this study, the effects of protein deficiency on Ca2+-ATPase activities were investigated. Twenty four Wistar rats (average weight 119±0.3 g) randomised into four groups of six (6) rats each were used for the study. Group A rats were treated with normal animal diet, group B were placed on protein-deficient diet, Group C were treated with protein deficient diet fortified with fish while Group D were administered with protein deficient diet fortified with Moringa oleifera leaves powder. They were maintained on respective diets for 6 weeks then sacrificed. The skeletal muscle, brain and liver were removed and homogenised. Vanadate is a known cellular Ca2+-ATPase inhibitor and was used to confirm the sensitivity of the enzyme in the homogenate. The protein concentration in each formulated diet was determined to be 9.71%, 2.19%, 23.92% and 31.54% for group A, B, C and D respectively. The enzyme’s activities were highest in group D and lowest in Group B. In the same line, the activities were also highest in the skeletal muscle and lowest in the liver of all the groups. The Km (ATP) and Vmax of the enzyme in all the studied organs were lowest in group D and highest in group B. The results indicate that protein deficiency reduces the activities of Ca2+-ATPase in the entire tissues’ cells and that M. oleifera leave powder is a better activator for the enzyme and therefore a good modulator of Ca2+-ATPase activities

Ammonia, a product of degradation of proteins and other nitrogen compounds is toxic at higher concentrations. Fisetin (a naturally occurring flavonoid) is known to exhibit a number of cytoprotective effects. In this study, chronotherapeutic effects of fisetin on ammonium chloride (AC) treated rats is aimed to establish the time point at which the maximum drug effect is achieved. Fisetin (50 mg/kg b.w oral) was administrated to rats at 06:00, 12:00, 18:00 and 00:00 h against AC (100 mg/kg.b.w. i.p) treated Wistar rats (180-200 g). Amelioration of pathophysiology of AC induced hyperammonaemic characteristics by fisetin at different time points were measured by assessing cellular architecture of liver and brain, lipid profile, lipid peroxidation products and antioxidants (superoxide dismutase, catalase, glutathione peroxidase and reduced glutathione) in serum, plasma, liver and brain. The normalization of lipid profile and markers of reactive oxygen species (ROS) homeostasis indicated the protective effect of fisetin. Fisetin administration at 00:00 h showed significant effects on these indices than at other time points (p<0.05; DMRT). This might be due to chronopharmacokinetic and chronopharmacodynamic properties of fisetin along with its action on altered rhythms of lipid profile and markers of ROS homeostasis.

The hydroglycolic extracts from Terminalia chebula Retzius, Terminalia bellerica Roxb and Rafflesia kerrii Meijer were investigated for total phenolic content (TPC), cytotoxicity, mutagenicity, antimutagenicity and antityrosinase for safety assessment as novel botanical-based cosmeceutical ingredients. These plant extracts showed TPC between 14.90 ± 0.02 and 112.40 ± 0.08 mg GAE g-1 of extract when using the Folin-Ciocalteu method. The cytotoxicity study revealed that the 50% cytotoxicity dose (CD50) towards normal mouse fibroblast L929 and mouse melanoma B16F10 cell lines was 5.43 ± 0.18 - 39.39 ± 0.14 mg mL-1 and 4.35 ± 0.33 - 58.23 ± 0.18 mg mL-1, respectively. In genotoxicity investigation, it was found that all extracts were not mutagenic at the concentrations up to 87.34 mg 0.1 mL-1 when tested with Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation (S9 microsomal fraction). The extracts were further tested for antimutagenic activity against 2-aminoanthracene (2-AA) and 2-(2-furyl)-3-(5-nitro-2-furyl) acrylamide (AF-2) which were used as the tested mutagens. Interestingly, all hydroglycolic extracts exhibited the inhibitory effect on the mutagenicity after being induced by 2AA and AF-2 in S. typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation. All plant extracts were further investigated for tyrosinase inhibitory activity. Results showed that all extracts possessed tyrosinase inhibitory activity with 50% inhibitory concentration values (IC50) of 1.27 ± 0.49 - 39.96 ± 0.21 mg mL-1. Overall studies including their antimutagenicity and antityrosinase activities suggest that the hydroglycolic extracts of these three plants may be used as potential candidates for skin-care cosmeceutical ingredients.

Combination of natural compounds and cytotoxic drugs represents a logical therapeutic approach for breast cancer. Brucine, a natural plant alkaloid is reported to possess cytotoxic, anti-inflammatory and anti-cancer activities. Gemcitabine is a nucleoside analog, commonly used in the treatment of several solid tumors, including breast cancer. In the present study we have examined the anticancer effect of brucine and gemcitabine on MDA MB-231 human breast cancer cells. Cell proliferation was assessed using MTT assay. Soft agar assay was used to evaluate the in-vitro clonogencity of MDA MB-231 cells. Cell migration was determined by in-vitro scratch assay and expression of p65 (NF-kB subunit) was evaluated by western blot analysis. Combination treatment with brucine and gemcitabine resulted in a significant inhibition of cell proliferation than either brucine or gemcitabine alone. Cells treated with combination of brucine and gemcitabine showed additive inhibition in colony formation and cell migration than treated with individual agents. The cells treated with brucine at 300 µM showed a significant decrease in p65-NF-kB expression but in combination treatment there was no additive inhibition of p65 expression compared to brucine treated cells. Overall, our results suggested that brucine in combination with gemcitabine showed supra-additive anticancer effects in MDA MB-231 cells and warrants further in-vivo studies in experimental animal models.