Histone Deacetylase Inhibitors (HDACi) Promote KLF5 Ubiquitination and Degradation in Basal-like Breast Cancer.

Basal-like breast cancer (BLBC) accounts for approximately 15% of all breast cancer cases, and patients with BLBC have a low survival rate. Our previous study demonstrated that the KLF5 transcription factor promotes BLBC cell proliferation and tumor growth. In this study, we demonstrated that the histone deacetylase inhibitors (HDACi), suberoylanilide hydroxamic acid (SAHA), and trichostatin A (TSA), increased KLF5 acetylation at lysine 369 (K369), downregulated KLF5 protein expression levels, and decreased cell viability in BLBC cell lines. HDACi target KLF5 for proteasomal degradation by promoting KLF5 protein ubiquitination. K369 acetylation of KLF5 decreases the binding between KLF5 and its deubiquitinase, BAP1. These findings revealed a novel mechanism by which HDACi suppress BLBC, and a novel crosstalk between KLF5 protein acetylation and ubiquitination.

Molecular investigation of possible relationships concerning bovine leukemia virus and breast cancer.

Worldwide, breast cancer has an eminent morbidity and mortality rate, as it is a neoplastic disease among females. The query of the prospective danger of bovine leukemia virus (BLV) to humans is an old but exceedingly topical focus of scientific debate. The objective of the current study was to determine the possible relationship between BLV and breast cancer. A total of 2710 formalin-fixed paraffin-embedded (FFPE) breast cancer samples were selected regardless of the age, ethnicity, or municipality origin of the subjects. The presence of BLV in human breast cancer was determined through nested PCR by amplifying tax and gag genes followed by partial sequencing. Homology was confirmed by using the online BLAST Tool. BLV genes were found to be positive in 26.8% (728/2710) of the samples from breast cancer patients and 10% (10/80) of the samples without cancer (negative control). The results indicated a correlation between the presence of the BLV gene and breast cancer (odds ratio = 0.3889; confidence interval = 1,18; p = 0.0029). The current findings suggest a possible link between BLV and human breast carcinoma. Therefore, screening cattle herds and milk products is suggested to reduce the viral transmission risk to humans.

Investigation of Chemopreventive and Antiproliferative Potential of Dicliptera roxburghiana.

CONTEXT: Carcinogenesis causes much human misery. It is a process involving multistage alterations. Medicinal plants are candidates for beneficial anticancer agents. OBJECTIVES: Investigation of anticancer proficiencies of the plant Dicliptera roxburghiana. MATERIAL AND METHODS: Crude extract and derived fractions were inspected for their inhibitory potential against nuclear factor KB (NFκB), nitric oxide synthase inhibition, aromatase inhibition and induction of quinone reductase 1 (QR 1). Antiproliferative activity was determined by using various cancer cell lines for example hormone responsive breast cancer cell line MCF-7, estrogen receptor negative breast cancer cell line MDA-MB-231, murine hepatoma cells Hepa 1c1c7, human neuroblastoma cells SK-N-SH and neuroblastoma cells MYCN-2. RESULTS: Ethyl acetate and n-butanol fractions of D. roxburghiana were strongly active against NFκB with IC50 of 16.6 ± 1.3 and 8.4 ± 0.7 µg/ml respectively with 100% survival. Chloroform fraction of the plant exhibited an induction ratio of 2.4 ± 0.09 with CD value of 17.7 µg/ml. Regarding the nitrite assay, the n-hexane fraction exhibited significant inhibition of NO activity with IC50 of 17.8 ± 1.25 µg/ml. The n-butanol fraction exhibited strong antiproliferative activity against IcIc-7 cell lines with IC50 values of 13.6 ± 1.91 µg/ml; against MYCN-2 a cytotoxic effect developed with dose dependence, with IC50 of 12.6 ± 1.24 µg/ml. In antiproliferative activity against SK-N-SH cell lines, chloroform, ethyl acetate and n-butanol fractions were efficiently active with IC50 values of 11.2 ± 0.84, 14.6 ± 1.71 and 16.3 ± 1.57 respectively. DISCUSSION AND CONCLUSION: It was demonstrated that various fractions of D. roxburghiana displayed appreciable anticancer characteristics and could be a potent source for the development of anticancer leads.

Phytochemical investigation and nephroprotective potential of Sida cordata in rat.

BACKGROUND: Plants are an efficient source of natural antioxidant against free radicals causing kidney damages. Sida cordata ethyl acetate fraction has been reported for strong in vitro antioxidant potency, previously. In the present study, our objective was to evaluate its in vivo antioxidant potency against CCl4 induced nephrotoxicity and investigates the bioactive phytochemicals by HPLC-DAD analysis. METHODS: Phytochemical analysis was performed by HPLC-DAD methodology. For in vivo study, 42 male Sprague-Dawley rats were treated with alternatively managed doses for 60 days. Group I animals were remained untreated. Group II animals were treated with vehicle (1 mL of olive oil) by intragastric route on alternate days. Group III was treated with 30% CCl4 (1 mL/kg b.w.) i.p. Group IV was treated with 30% CCl4 (1 mL/kg b.w.) i.p and silymarin intragastric. Group V and VI rats were treated with 30% CCl4 and SCEE (150 and 300 mg/kg b.w., respectively) intragastric. Group VII animals were treated with SCEE (300 mg/kg b.w.) intragastrically. Blood parameters, Serum proteins and urine profile were investigated. Activities of tissue enzyme i.e. catalase, peroxidase, superoxide dismutase, glutathione-S-transferase, glutathione reductase, GSH and γ-GT were evaluated. Histopathological observations, total protein contents, lipid peroxidation, DNA damage and relative weight were also analyzed. RESULTS: Gallic acid, catechin and caffeic acid were identified in SCEE fraction by HPLC-DAD. Decrease in the count of red blood cells, neutrophils, eosinophils and concentration of hemoglobin whereas increase in lymphocyte count and estimation of sedimentation rate (ESR) with 1 mL CCl4 (30% in Olive oil) administration (30 doses in 60 days) was restored dose dependently with co-treatment of SCEE (150 and 300 mg/kg b.w.). Treatment of rats with CCl4 markedly (P < 0.01) increased the count of urinary red blood cells and leucocytes, concentration of urea, creatinine and urobilinogen and specific gravity whereas creatinine clearance was reduced. Serum level of total protein, albumin, globulin, nitrite, creatinine and blood urea nitrogen (BUN) was significantly increased (P < 0.01) by CCl4 treatment. The activity of antioxidant enzymes; catalase, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase and glutathione reductase and content of reduced glutathione was decreased (P < 0.01) significantly. However, increased concentration (P < 0.01) of thiobarbituric acid reactive substances and histopathological injuries were noticed in the renal tissues of rats after the treatment with CCl4. Co-administration of SCEE, dose dependently, protected the alterations in the studied parameters of rats at 150 and 300 mg/kg b.w. The present study revealed that SCEE could be used as a possible remedy for renal toxicity abnormalities. CONCLUSION: These results are an evidence of the renal protective role of S.cordat ethyl acetate fraction against CCl4 induced nephrotoxicity in rats which may be due to its antioxidant compounds.

Pistacia chinensis: Strong antioxidant and potent testicular toxicity amelioration agent.

OBJECTIVES: To evaluate in vitro and in vivo antioxidant potency of Pistacia chinensis (P. chinensis) bark and leaves extracts along with its protective role against CCl4 induced toxicity in testis of the rat. METHODS: Various in vitro models such as DPPH, ABTS, hydrogen peroxide, superoxide, hydroxyl and nitric oxide scavenging activities, anti-lipid peroxidation activity, phospho-molybdenum activity, ß carotene bleaching assay was used for analysis of antioxidant potential. Experimental groups for in vivo study were: Group Ⅰ (control) untreated, Group Ⅱ (Vehicle control), Group Ⅲ (1 mL/kg b.w 30% CCl4), Group Ⅳ (1 mL/kg b.w CCl4 + Silymarin), Group Ⅴ (200 mg/kg b.w PCBE + CCl4), Group Ⅵ (400 mg/kg b.w PCBE + CCl4) and Group Ⅶ (400 mg/kg b.w PCBE alone). RESULTS: In vitro antioxidant assays displayed significant results and the highest activity was not specified to a specific extract. However, ethyl acetate extract of bark (PCBE) showed highest results in most of the antioxidant assays i.e. beta-carotene bleaching, hydroxyl radical scavenging, ABTS, lipid peroxidation and superoxide radical scavenging activity. On this base, this fraction was selected for in vivo antioxidant experiment. Testis tissues were analyzed to observe the protective effects of PCBE on antioxidant enzymes; catalase, superoxide dismutase, peroxidase, glutathione-S-transferase, glutathione reductase, glutathione peroxidase and quinone reductase activities and glutathione (GSH) as well as nitrite content. Profile of plasma testosterone was also compared to various treatments. Observation suggests a protective role of P. chinensis against CCl4 induced toxicity. CONCLUSIONS: It is concluded that some bioactive antioxidants of P. chinensis bark might be a good source to isolate the potent antioxidant components.

Increase of glutathione, testosterone and antioxidant effects of Jurenia dolomiaea on CCl4 induced testicular toxicity in rat.

BACKGROUND: Root of Jurenia dolomiaea is used traditionally in various disorders involving oxidative injuries i.e. rheumatism, gout and as stimulant. Earlier we have investigated in vitro antioxidant and DNA protective ability. In this investigation we have evaluated protective potential of J. dolomiaea root against the oxidative injuries induced with carbon tetrachloride (CCl4) in testes of rat. METHODS: Dried roots of J. dolomiaea were powdered and extracted with 95% methanol and residue was fractionated in escalating polarity of solvents. On the basis of potent antioxidant ability; the ethyl acetate fraction (JDEE) was selected to evaluate the in vivo antioxidant activity against CCl4 induced oxidative stress in rat. Sprague Dawley male rats (42) were equally divided in to 7 groups: control, vehicle control, JDEE (400 mg/kg; p.o.) alone, CCl4 (I ml/kg; 1:10 v/v in olive oil) alone, JDEE (200 mg/kg, 400 mg/kg) with CCl4, and silymarin (200 mg/kg) with CCl4 on alternate days for 60 days. Testes samples were investigated for antioxidant enzymes, biochemical markers and histopathology while the serum samples were analyzed for the testosterone level. RESULTS: Administration of CCl4 to rats depleted the activity level of antioxidant enzymes viz.; CAT, POD, SOD, GST, GPx, and GR, and the concentration of protein and GSH while enhanced the level of lipid peroxides (TBARS), H2O2 and nitrite in testes samples of rat. Concentration of testosterone in serum of rat decreased with CCl4 treatment. Co-treatment of silymarin and the JDEE (200 mg/kg, 400 mg/kg) lessened the toxic effects of CCl4 and reversed the level of these parameters towards the control group. An admirable increase (P < 0.05) in the level of GSH in testes, testosterone in serum and thickness of germinal layers in testes with JDEE (400 mg/kg) alone was recorded. Histopathological observation of testes samples endorsed the alterations induced with different treatments. CONCLUSIONS: JDEE co-treatment to rats ameliorated the toxic effects of CCl4 in testes samples. Enhanced level of GSH, thickness of germinal layers in testes and testosterone in serum with JDEE (400 mg/kg) treatment alone to rats demanded the evaluation of JDEE for sexual behavior.

Evaluation of phytochemicals, antioxidant activity and amelioration of pulmonary fibrosis with Phyllanthus emblica leaves.

BACKGROUND: In the present study the antioxidant potential of a methanol extract of Phyllanthus emblica leaves (PELE) was determined by in vitro methods as well as by an in vivo animal model, along with HPLC-DAD screening for phyto-constituents. METHODS: The in vitro antioxidant potential of PELE was assessed by scavenging of DPPH, nitric oxide and anti-lipid peroxidation assays. For in vivo evaluation, a 60-day experimental plan was followed in which Sprague Dawley rats were administered with 1 mL/kg of CCl4 (CCl4 : DMSO + Olive oil; 30 % v/v) alone or with different doses of PELE (200, 400 mg/kg p.o.). Silymarin (100 mg/kg) as standard drug was also administered to CCl4 treated rats. HPLC-DAD analysis was performed to quantify polyphenolic phytochemicals. RESULTS: PELE exhibited an appreciable in vitro antioxidant activity and scavenged the DPPH radical (IC50 = 39.73 ± 2.12 µg/mL) and nitric oxide (IC50 = 39.14 ± 2.31 µg/mL) while for anti-lipid peroxidation moderate antioxidant activity was noticed. Reduced levels of antioxidant enzyme activities viz., superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and reduced glutathione (GSH) whereas enhanced levels of total extractable proteins, lipid peroxides (TBARS), nitrite and H2O2 were induced by CCl4 administration in lungs of rat. Co-administration of PELE to rats exhibited a dose dependent decline in the oxidative injuries induced in these parameters. Histopathological damages such as disrupted alveoli, infiltration of macrophages and modified architecture of Clara cells was reversed to the normal state with co-administration of PELE. HPLC-DAD analysis indicated the presence of gallic acid, rutin, kaempferol and caffeic acid in the PELE. CONCLUSION: The findings of this study demonstrate that presence of polyphenolics and other active constituents in PELE might play a significant role in repairing the pulmonary damages instigated with CCl4.

Proficiencies of Artemisia scoparia against CCl4 induced DNA damages and renal toxicity in rat.

BACKGROUND: Artemisia scoparia is traditionally used in the local system of medicine in kidney disorders. This study aimed at scrutinizing the nephroprotective prospective of A. scoparia methanol extract against carbon tetrachloride (CCl4) provoked DNA damages and oxidative stress in kidneys of rat. METHODS: Dried aerial parts of A. scoparia were powdered and extracted with methanol to obtain the viscous material (ASM). Sprague Dawley male rats (42) were grouped (7) having 6 rats in each. Group I remained untreated and Group II treated intraperitoneally (i.p) with DMSO + olive oil (1 ml/kg body weight (bw). Rats of Group III - VI were treated with CCl4 (1 ml/kg bw; i.p 30 % v/v in olive oil). Animals of Group IV were co-administered with 100 mg/kg bw of silymarin whereas rats of Group V and VI with 150 mg/kg bw and 300 mg/kg bw of ASM at an interval of 48 h for four weeks. Animals of Group VII were administered with ASM (300 mg/kg bw) alone. DNA damages were investigated with comet assay in renal tissues while the oxidative injuries were estimated in serum and renal tissues. RESULTS: Co-administration of ASM to rats significantly reduced the DNA damages at 300 mg/kg dose as indicated in comet length (40.80 ± 2.60 µm), head length (34.70 ± 2.21 µm), tail length (7.43 ± 1.24 µm) and DNA content in head (88.03 ± 2.27 %) to that of CCl4 for comet length (63.16 ± 2.11 µm), head length (23.29 ± 1.50 µm), tail length (39.21 ± 2.81 µm) and DNA content of head (74.81 ± 2.18 %) in renal cell's nuclei. Increased level of urea, creatinine, bilirubin, blood urea nitrogen whereas decreased concentration of proteins in serum of CCl4 treated animals were restored towards the normal level with co-administration of ASM. CCl4 injection in rats decreased the activity level of CAT, POD, SOD, GST and γ-GT and GSH contents while elevated levels of TBARS, H2O2 and nitrite contents were observed in renal tissues. A noteworthy retrieval of all these parameters and the altered histopathological observations was notified near to the normal values after treatment with both the doses of ASM. CONCLUSION: Results obtained suggested the therapeutic role of ASM in oxidative stress related disorder of kidneys.

Antitumor characterization of various fractions of Launaea procumbens.

The antitumor activity of various six fractions of Launaea procumbens were studied using the potato disk bioassay technique. Three concentrations (10, 100, and 1000 ppm) of all these six fractions were used for characterization of antitumor activity. Among these fractions, methanolic fraction exhibited significant inhibition of crown gall tumors caused by Agrobacterium tumefaciens followed by butanolic fraction. These results revealed that methanolic and butanolic fractions contain bioactive constituents responsible for inhibition of tumor. Further purification and characterization are in progress in our laboratory.

Studies on phytochemical, antioxidant, anti-inflammatory and analgesic activities of Euphorbia dracunculoides.

BACKGROUND: Plants provide an alternative source to manage various human disorders due to diverse metabolites. Euphorbia dracunculoides of family Euphorbiaceae is used by local practitioners in rheumatism, epilepsy, edema, snake bite, warts and also possesses diuretic and purgative effects. The present study evaluated the antioxidant, anti-inflammatory and analgesic activities of various extracts of E. dracunculoides. Further, phytochemical constituents of the leading extracts were also investigated. METHODS: Dry powder of E. dracunculoides was extracted with n-hexane (EDH), acetone (EDA), ethanol (EDE), ethanol + water (1:1) (EDEW) and methanol (EDM) and screened for phytochemical classes, total phenolic (TPC) and flavonoid content (TFC). Antioxidant effects of the extracts were manifested by in vitro multidimensional assays. The anti-inflammatory and analgesic activities of the extracts were evaluated through carrageenan induced paw edema and hot plate test in rat. In addition, GC-MS analysis of EDH and HPLC-DAD analysis of EDEW was carried out to determine the presence of active constituents. RESULTS: Qualitative analysis of various extracts of E. dracunculoides assured the existence of tannins and coumarins while presence of anthraquinones and anthocyanins was not traced in these extracts. Maximum quantity of TPC and TFC was recorded in EDEW followed by EDE. EDEW and EDE showed significant antioxidant activities with therapeutic potential against hydroxyl and phosphomolybdate radicals, ß-carotene bleaching assay and in reducing of iron while moderate to low scavenging abilities were recorded for DPPH, nitric oxide and for iron chelation. During anti-inflammatory activity after 4 h of drug administration the 300 mg/kg body weight dose of EDH (68.660 ± 10.502%) and EDE (51.384 ± 8.623%) exhibited strong anti-inflammatory activity and reduced the carrageenan-induced paw edema in rat as compared to standard drug diclofenac sodium (78.823 ± 6.395%). Treatment of rats with EDH (70.206 ± 5.445%) and EDE (56.508 ± 6.363%) after 90 min showed significant increase in percent latency time in hot plate test as compared to morphine (63.632 ± 5.449%) treatment in rat. GC-MS analysis of EDH indicated the presence of 30 compounds predominantly of steroids and terpenoids. HPLC-DAD analysis against known standards established the presence of rutin, catechin, caffeic acid and myricetin in EDEW. CONCLUSION: Our results suggest that presence of various polyphenolics, terpenoids and steroids render E. dracunculoides with therapeutic potential for oxidative stress and inflammation related disorders.

Amelioration of carbon tetrachloride-induced pulmonary toxicity with Oxalis corniculata.

This research work was planned to investigate the antioxidant potential of methanolic crude extract of Oxalis corniculata (OCME) against lung injuries initiated by carbon tetrachloride (CCl4) in rats at histological and biochemical level. A total of 42 female Sprague Dawley rats were randomly distributed in to seven groups and each group comprised of six rats. Experiment was completed in 22 days (10 doses at alternate days). Group I was not treated (control rats), while group II was administered with vehicles (olive oil and dimethyl sulfoxide), groups III, IV, and V were treated with 1 ml kg(-1) body weight (b.w.) of CCl4 (20% in olive oil). Group III received only CCl4, whereas groups IV and V were administered with 100 and 200 mg kg(-1) b.w. of OCME, respectively. Group VI was administered with OCME (200 mg kg(-1) b.w.) alone. Group VII was treated with sylimarin (50 mg kg(-1) b.w.). CCl4 enhanced the lipid peroxidation while reduced the glutathione in lung samples. Activities of antioxidant enzymes, catalase, peroxidase, superoxide dismutase, and glutathione-S-transferase decreased in lung homogenates with CCl4. Treatment of CCl4 induced deleterious changes in the microanatomy of lungs by rupturing the alveolar septa, thickening of alveolar walls, and damaging the cells with subsequent collapse of blood vessels due to the accumulation of degenerated blood cells. OCME, dose dependently, prevented the alterations in these parameters. These results suggest that OCME protected the lungs due to its intrinsic properties by scavenging of free radicals generated by CCl4.

HPLC-DAD analysis, antioxidant potential and anti-urease activity of Asparagus gracilis collected from District Islamabad.

BACKGROUND: Asparagus gracilis subspecie of Asparagus capitatus Baker, is described as food and medicine for various ailments. In this study we investigated, its phenolic constituents, in vitro antioxidant potential against various free radicals and anti-urease potential. METHODS: Asparagus gracilis aerial parts collected from District Islamabad, Pakistan were extracted with crude methanol which was further fractionated into n-hexane, ethyl acetate, n-butanol and aqueous fraction. Total phenolic and flavonoid contents were estimated for extract and all the derived fractions. Diverse in vitro antioxidants assays such as DPPH, H2O2, •OH, ABTS, ß-carotene bleaching assay, superoxide radical, lipid peroxidation, reducing power, and total antioxidant capacity were studied to assess scavenging potential. Antiurease activity of methanol extract and its derived fractions was also investigated. HPLC-DAD analysis of crude methanol extract was performed by using different phenolic standards. RESULTS: Ethyl acetate fraction expressed maximum content of flavonoids (240.6 ± 6.1 mg RE/g dry sample), phenolics (615 ± 13 mg GAE/g dry sample) and best antioxidant potential among different fractions of crude methanol extract. Hydrogen peroxide assay and hydroxyl, supeoxide, nitric oxide free radicals antioxidant assays as well as beta carotene assay showed significant correlation with flavonoid content while hydrogen peroxide, ABTS and lipid peroxidation assay displayed significant correlation with phenolic content. HPLC analysis showed the presence of important phenolics i.e. catechin (4.04 ± 0.02 µg/mg sample), caffeic acid (0.89 ± 0.003 µg/mg sample), rutin (24.58 ± 0.1 µg/mg sample), myricetin (1.13 ± 0.07 µg/mg sample) and quercetin (14.91 ± 0.09 µg/mg sample). Ethyl acetate fraction expressed lowest IC50 in antiurease activity. Correlation analysis of antiurease activity expressed significant correlation with flavonoids (P < 0.004) and phenolics (P < 0.02) proposing multipotent activity of fractions. CONCLUSION: These results revealed the presence of some bioactive compound in the ethyl acetate fraction having both antioxidant as well as antiurease potential.

Estimation of flavoniods, antimicrobial, antitumor and anticancer activity of Carissa opaca fruits.

BACKGROUND: Carissa opaca Stapf ex Hanes fruits is traditionally used in the treatment of asthma, hepatitis and microbial infections. The present study was arranged to investigate the antimicrobial, cytotoxic and antitumor activity of various fractions of C. opaca extract and its bioactive metabolites responsible for that activity. METHODS: To characterize various fractions of C.opaca antibacterial, antifungal, cytotoxic and antitumor assays are used. Eight strains of bacteria including Bacillus subtilis, Enterobactor aerogenes, Escherichia coli, Klebsiella pneumoniae, Micrococcus luteus, Pseudomonas aeroginosa, Salmonella typhy, and Staphylococcus aureus and four strains of fungal viz: Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger and Fusarium solani are used. Brine shrimps and potato dics are used for anticancer and antitumor potency of extract. High performance liquid chromatography (HPLC) is utilized for determination of bioactive metabolites responsible for the activity. RESULTS: HPLC chromatogram revealed the presence of orientin, isoquercetin, myricetin and apigenin. Various fractions of C. oapca showed significant antibacterial, antitumor and anticancer activity. In case of C. opaca fruit inhibition growth of Aspergillus niger was ranged between 23.2 ± 1.36% to 43.3 ± 2.39%, Aspergillus flavus ranged between 27.6 ± 1.39% to 65.6 ± 3.44%, Aspergillus fumigatus ranged between 13.2 ± 1.00% to 52.4 ± 1.54% and Fusarium solani ranged between 10.5 ± 1.02% to 14.6 ± 1.74%. CONCLUSION: It can be concluded that, various fractions of C.opaca are accessible source of ethno pharmacy as they are consumed in different areas of Pakistan with ultimate health compensations.

Investigation on flavonoid composition and anti free radical potential of Sida cordata.

BACKGROUND: Sida cordata, a member of Family Malvaceae is used in folk medicine for various ailments including liver diseases. In this study we investigated, its flavonoid constituents, in vitro antioxidant potential against different free radicals and hepatoprotection against carbon tetrachloride (CCl4)-induced liver damage in rat. METHODS: Dried powder of S. cordata whole plant was extracted with methanol and the resultant (SCME) obtained was fractionated with escalating polarity to obtain n-hexane fraction (SCHE), ethyl acetate fraction (SCEE), n-butanol fraction (SCBE) and the remaining soluble portion as aqueous fraction (SCAE). Diverse in vitro antioxidants assays such as DPPH, H2O2, •OH, ABTS, ß-carotene bleaching assay, superoxide radical, lipid peroxidation, reducing power, and total antioxidant capacity were studied to assess scavenging potential of methanol extract and its derived fractions. On account of marked scavenging activity SCEE was selected to investigate the hepatoprotective potential against CCl4 induced toxicity in Sprague-Dawley male rats by assessing the level of serum markers (alkaline phosphatase, alanine transaminase, aspartate transaminase, lactate dehydrogenase, bilirubin, and γ-glutamyltransferase) and of liver antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione-S-transfers (GST), glutathione reductase (GSR), glutathione peroxidase (GSH-Px), and reduced glutathione (GSH) and lipid peroxidation (TBARS). Histology of the liver was performed to study alteration in histoarchitecture. Existence of active flavonoids was established by thin layer chromatographic studies. RESULTS: Considerable amount of flavonoid and phenolic contents were recorded in the methanol extract and its derived fractions. Although the extract and all its derived fractions exhibited good antioxidant activities however, the most distinguished scavenging potential was observed for SCEE. Treatment of SCEE decreased the elevated level of serum marker enzymes induced with CCl4 administration whereas increased the activity of hepatic antioxidant enzymes (CAT, SOD, POD, GST, GSR and GSH-Px). Hepatic concentration of GSH was increased while lipid peroxidation was decreased with SCEE administration in CCl4 intoxicated rats. Presence of apigenin with some unknown compounds was observed in SCEE by using thin layer chromatography. CONCLUSIONS: These results revealed the presence of some bioactive compound in the ethyl acetate fraction, confirming the utility of S. cordata against liver diseases in folk medicine.

Antitumor, antioxidant and antimicrobial studies of substituted pyridylguanidines.

A series of N-pivaloyl-N'-(alkyl/aryl)-N''-pyridylguanidine of general formula C4H9CONHC(NR¹R²)NPy have been synthesized and characterized using elemental analysis, FT-IR, multinuclear NMR spectroscopy, and in the case of compounds 7 and 11, by single crystal X-ray diffraction (XRD). The synthesized guanidines were tested for antitumor activities against potato tumor, and showed excellent inhibition against Agrobacterium tumefaciens (AT10)-induced tumor. The antioxidant and antimicrobial activities of these new compounds against various bacterial and fungal strains were also investigated.

Assessment of phytochemicals, antimicrobial and cytotoxic activities of extract and fractions from Fagonia olivieri (Zygophyllaceae).

BACKGROUND: In Pakistan Fagonia olivieri (Zygophyllaceae) is commonly used in the indigenous system of medicine for treatment of conditions like diabetes, cancer, fever, asthma, toothache, stomach troubles and kidney disorders. This study evaluated the crude methanol extract of F. olivieri (FOM) and its derived fractions for their antimicrobial and cytotoxic activities as well as the classes of phytochemical. METHODS: Dried powder of whole plant of F. olivieri was extracted with methanol (FOM) and the resultant was fractionated to give n-hexane fraction (FOH), chloroform fraction (FOC), ethyl acetate fraction (FOE), n-butanol fraction (FOB) and residual aqueous fraction (FOA). Methanol extract and its derived fractions were subjected to phytochemical screening using standard procedures. Also the extract and fractions were assayed for antibacterial, antifungal and cytotoxic activities using agar well diffusion technique, agar tube dilution method and brine shrimps lethality test, respectively. RESULTS: The results obtained for phytochemical analysis indicate the presence of saponins and alkaloids in all the tested extract and fractions while anthraquinones were not detected. The results showed that all the bacterial strains tested in this study were susceptible to at least one of the fractions tested. However, FOE and FOB were the best antibacterial fractions and showed antibacterial activity against maximum number of bacterial strains. The results showed that Escherichia coli was the most sensitive bacterium while Bordetella bronchiseptica and Enterobacter aerogenes were less susceptible against various fractions. Maximum percent inhibition for growth was recorded for the fungus Aspergillus flavus with FOE whereas growth of Aspergillus fumigatus and Fusarium solani was inhibited by FOM and its all derived fractions. Minimum LC50 (24.07 mg/L) for brine shrimp assay was recorded for FOE followed by LC50 of FOC (26.1 mg/L) and FOB (30.05 mg/L) whereas maximum LC50 was exhibited by FOH (1533 mg/L). CONCLUSION: These results indicated the use of F. olivieri to treat infections with emphasis to isolate and characterize the active principle responsible for antibacterial, antifungal and cytotoxic activities and its exploitation as therapeutic agent.

Synthesis, characterization, biological screenings and interaction with calf thymus DNA of a novel azomethine 3-((3,5-dimethylphenylimino)methyl)benzene-1,2-diol.

The novel azomethine, 3-((3,5-dimethylphenylimino)methyl)benzene-1,2-diol (HL) was synthesized and characterized by elemental analysis, FT-IR, (1)H, (13)C NMR spectroscopy and single crystal analysis. The title compound has been screened for its biological activities including enzymatic study, antibacterial, antifungal, cytotoxicity, antioxidant and interaction with CTDNA, and showed remarkable activities in each area of research. The titled compound interacts with DNA via two binding modes: intercalation and groove binding. In intercalation the compound inserts itself into the base pairs of DNA and the compound-DNA complex is stabilized by π-π stacking. Interaction via groove binding may be due to hydrogen bonding to bases, typically to N3 of adenine and O2 of thymine. The synthesized compound was also found to be an effective antioxidant of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and gives percent inhibition (%I) of 90.7 at a concentration level of 31.3µg/mL.