Genomic diversity and population structure of three autochthonous Greek sheep breeds assessed with genome-wide DNA arrays.

In the present study, genome-wide genotyping was applied to characterize the genetic diversity and population structure of three autochthonous Greek breeds: Boutsko, Karagouniko and Chios. Dairy sheep are among the most significant livestock species in Greece numbering approximately 9 million animals which are characterized by large phenotypic variation and reared under various farming systems. A total of 96 animals were genotyped with the Illumina's OvineSNP50K microarray beadchip, to study the population structure of the breeds and develop a specialized panel of single-nucleotide polymorphisms (SNPs), which could distinguish one breed from the others. Quality control on the dataset resulted in 46,125 SNPs, which were used to evaluate the genetic structure of the breeds. Population structure was assessed through principal component analysis (PCA) and admixture analysis, whereas inbreeding was estimated based on runs of homozygosity (ROHs) coefficients, genomic relationship matrix inbreeding coefficients (FGRM) and patterns of linkage disequilibrium (LD). Associations between SNPs and breeds were analyzed with different inheritance models, to identify SNPs that distinguish among the breeds. Results showed high levels of genetic heterogeneity in the three breeds. Genetic distances among breeds were modest, despite their different ancestries. Chios and Karagouniko breeds were more genetically related to each other compared to Boutsko. Analysis revealed 3802 candidate SNPs that can be used to identify two-breed crosses and purebred animals. The present study provides, for the first time, data on the genetic background of three Greek indigenous dairy sheep breeds as well as a specialized marker panel that can be applied for traceability purposes as well as targeted genetic improvement schemes and conservation programs.

Inhibition of lipoxygenase (LOX) and anticancer activity caused by gold(I) mixed ligands complexes of triphenylphosphine and thioamides.

Four mixed ligand gold(I) complexes with the thioamides 2-mercapto-thiazolidine (mtzdH), 2-mercapto-benzothiazole (mbztH) and 5-chloro-2-mercapto-benzothiazole (ClmbztH) and triphenylphosphine (tpp) of formulae [Au(tpp)Cl] (1) [Au(tpp)(mtzd)] (2), [Au(tpp)(mbzt)] (3) and [Au(tpp)(Clmbzt)] (4), already known, were used to study their mechanism of inhibition activity towards the catalytic oxidation of linoleic acid to hydroperoxylinoleic acid by the enzyme lipoxygenase (LOX), kinetically and theoretically. The results are compared to those of cisplatin. In addition, the anticancer cell screening results against leimyosarcoma (LMS) cells have shown that 2-4 complexes were more active than cisplatin. The uptake of complexes in LMS cells were also studied with electrospray ionisation mass spectrometry spectroscopy.

Structural properties, cytotoxicity, and anti-inflammatory activity of silver(I) complexes with tris(p-tolyl)phosphine and 5-chloro-2-mercaptobenzothiazole.

The synthesis and characterization of the silver(I) chloride complex of formula {[AgCI(CMBZT)(TPTP)(2)] . (MeOH)} (1) (CMBZT = 5-chloro-2-mercaptobenzothiazole, TPTP = tris(p-tolyl)phosphine) is described. Also the structure of the hydrate derivative {[AgCI(TPTP)(3)] . (0.5 . H(2)O)} (2) of the corresponding known anhydrous silver complex (Zartilas et al., 2009), and the polymorph 3 of the known [AgI(TPTP)(3)] complex (Zartilas et al., 2009) were determined and compared with the known ones. In addition, the structure of the known one silver(I) cluster {[AgI(TPTP)](4)} (4) (Meijboom et al., 2009) was re-determined at 120(2) K and possible Ag-Ag interactions were analyzed. The compounds 1-4 were characterized by X-ray crystallography at r.t (1) and 120 K (2-4). All these complexes and {[(Et(3)NH)(+)](2) . [Ag(6)(mu(3)-Hmna)(4)(mu(3)-mna)(2)](2-) . (DMSO)(2) . (H(2)O)} (5) (Hmna = 2-mercaptonicotinic acid) were evaluated for cytotoxic and anti-inflammatory activity. The in vitro testing of cytotoxic activity of 1-5 against leiomyosarcoma cancer cells (LMS), were evaluated with Trypan Blue and Thiazolyl Blue Tetrazolium Bromide or 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) assays. The flow cytometry assay for complex 1 and showed that at 15 muM of 1, 62.38% of LMS cells undergo apoptosis, while 7% of LMS cells undergo cell necrosis. The antitumor activity of 3 is comparable with that of its reported polymorph (Zartilas et al., 2009). The anti-inflammatory, activity of complexes 1-3 and 5 was also studied. The activity towards cell viability was 2 > 3 > 5 > 1 > 4, while the order of the inhibitory activity in cell growth proliferation follows the order, 2 > 3 > 1 > 4 > 5. The anti-inflammatory activity on the other hand is 1 > 2 > 5 > cdots, three dots, centered >3.

Synthesis, structural characterization and biological study of new organotin(IV), silver(I) and antimony(III) complexes with thioamides.

An overview of our work on the synthesis and biological activity of a series of tin(IV), silver(I) and antimony(III) complexes with thioamides is reported. Organotin(IV) complexes of formulae (n-Bu)2Sn(MBZT)2 (1), Me2Sn(CMBZT)(2) (2), {(Ph3Sn)2(MNA) (Me2CO)} (3), Ph3Sn(MBZT) (4), Ph3Sn(MBZO) (5), Ph3Sn(CMBZT) (6), Ph2Sn(CMBZT)2 (7) and (n-Bu)2Sn(CMBZT)2 (8), Me2Sn(PMT)2 (9), (n-Bu)2Sn(PMT)2 (10), Ph2Sn(PMT)2 (11), Ph3Sn(PMT) (12) {where MBZT=2-mercapto-benzothiazole, CMBZT=5-chloro-2-mercapto-benzothiazole, H2MNA=2-mercapto-nicotinic acid, MBZO=2-mercapto-benzoxazole and PMTH=2-mercapto-pyrimidine} were characterized by spectroscopic (NMR, IR, Mossbauer, etc.) and X-ray diffraction techniques and their influence on the peroxidation of oleic acid was studied. They were found to inhibit strongly the peroxidation of linoleic acid by the enzyme lipoxygenase. In addition, organotin(IV) complexes were found to exhibit stronger cytotoxic activity in vitro, against leiomyosarcoma cells, than cisplatin. The antiproliferative activity of the organotin complexes studied, against leiomyosarcoma cells follow the same order of LOX activity inhibition. This is, 3>>12>7>6 approximately 8 approximately 10>5 approximately 4>>2>9. Thus, among organotin(IV)-CMBZT complexes, 7 exhibits higher activity than the others and this is explained by a free radical mechanism, as it is revealed by an EPR study. The results are compared with the corresponding ones found for the silver(I) complexes of formulae complexes {[Ag6(mu3-HMNA)4(mu3-MNA)2](2-).[(Et(3)NH)+]2.(DMSO)2.(H2O)} (13), {[Ag4Cl4(mu3-STHPMH2)4]n} (14), {[Ag6(mu2-Br)6(mu2-STHPMH2)4(mu3-STHPMH2)2]n} (15), {[Ag4(mu2STHPMH2)6](NO3)4}(n) (16), {[AgCl(TPTP)]4} (17), [AgX(TPTP)3] with X=Cl (18), Br (19), I (20) (where STHPMH2=2-mercapto-3,4,5,6-tetrahydro-pyrimidine, TPTP=tri(p-toly)phosphine) and those of antimony(III) complexes {[SbCl2(MBZIM)4](+).Cl(-).2H2O.(CH3OH)} (21), {[SbCl2(MBZIM)4]+.Cl(-).3H2O.(CH3CN)} (22), [SbCl3(MBZIM)2] (23), [SbCl3(EMBZIM)2] (24), [SbCl3(MTZD)2] (25), {[SbCl3(THPMT)2]} (26) and {[Sb(PMT)3].0.5(CH3OH)} (27) (where MBZIM is 2-mercapto-benzimidazole, EMBZIM=5-ethoxy-2-mercapto-benzimidazole and MTZD is 2-mercapto-thiazolidine), which they have characterized with similar techniques as in case of organotin(IV) complexes. Silver(I) and antimony(III) complexes were found to be cytotoxic against various cancer cell lines.

Enhancement of the antineoplastic effect of anticarcinogens on benzo[a]pyrene-treated Wistar rats, in relation to their number and biological activity.

Naturally occurring anticarcinogens, such as vitamins C and E, and the microelement selenium were found to inhibit the induction of benzo[a]pyrene-induced malignant tumors in Wistar rats to various extends. The antineoplastic effect of the tested anticarcinogens is gradually increased according to the number of inhibitors selected. To date the maximum action against malignancy is manifested by use of the above three inhibitors. In the group of rats receiving vitamins C, E and selenium, the prolongation of life induced by adding more than one anticarcinogen to the treatment regime reached, and in some cases surpassed, the normal life expectancy of the rats. It is expected that by adding even more anticarcinogens, the antineoplastic potency (Ap) of the inhibitors will be further improved. These results encouraged us to conduct a clinical trial in terminal human cancer cases, in conjunction with the usual treatments of surgery or chemotherapy and irradiation.

Dose-related preventive and therapeutic effects of antioxidants-anticarcinogens on experimentally induced malignant tumors in Wistar rats.

A combination of antioxidants-anticarcinogens, consisting of vitamins C and E, selenium and 2-mercaptopropionyl glycine (2-MPG), was administered orally for the prevention (PRG) and treatment (TRG) of benzo(a)pyrene (BaP)-induced malignant tumors (leiomyosarcomas), in Wistar rats. In order to evaluate dose-related effects, a low dose vitamin (0.15 g/kg b.w. per day of vit.C and 0.05 g/kg b.w. per day of vit.E) and a high dose (1.5 g/kg b.w. per day of vit.C and 0.5 g/kg b.w. per day of vit.E) combination was administered, in prevention and treatment groups. Selenium was administered in doses of 2 microg/kg b.w. per day and 2-MPG in 15 mg/kg b.w. per day, in all groups. Daily estimations of 24 h urine volume levels of thiobarbituric acid reacting substances (MDA) were performed in 20 animals, divided into a control group, a BaP-injected group, a tricapryline-injected group and a BaP-injected and treated by the low dose combination group. Results revealed that the low dose combination failed to exert any beneficial effect on mean survival time of animals treated either preventitively or therapeutically. An increased number of animals bearing a second (lung) tumor was, in addition, found in autopsy and histological examination in the low dose combination (PRG and TRG) and the high dose TRG groups. The high dose combination groups manifested a significant prolongation of the mean survival time of animals; complete remission of tumors developed in 16.8% of the animals in the treatment group and a 5.2% prevention of tumor formation in the preventive group, without any evidence of an increased number of double tumor formation in the PRG group. Urine MDA increased significantly in animals injected by BaP during the first 10 days and since the 90th day (formation of palpable tumors) after injection, in relation to control and tricapryline-injected groups. Complete prevention of urine MDA-increased values was obtained in BaP-injected and treated by the low dose combination animals. Results indicate that high doses (megadoses) of the antioxidant-anticarcinogen vitamins C and E in combination with carefully selected other antioxidants possessing supplementary actions, are probably needed in order to achieve a sufficient prevention and treatment of malignant diseases.

Comparison of the therapeutic effects of two vanadium complexes administered at low dose on benzo[a]pyrene-induced malignant tumors in rats.

The antitumor effects of low dose administration of the vanadium(III) complexes with L-cysteine (complex 1) and N-(2-mercaptopropionyl)-glycine (complex 2) were compared on benzo[a]pyrene (BaP)-induced tumors in Wistar rats. Male Wistar rats, injected with 10.0 mg of BaP, were divided into one control (C-G) and two treatment (TR-G) groups of 17 animals each. Animals of the first treatment group were administered complex 2 (TR-2 group) and those of the second group were administered complex 1 (TR-1 group) at doses of 100 microg of vanadium per os daily, starting from the day a palpable tumor was developed till their death. BaP injection induced a 100% tumor (leiomyosarcomas) development in the animals of all groups. Administration of complex 1 to the animals resulted in a significant prolongation of the mean survival time, a complete remission of 17.6% of the tumors developed, a significant reduction of the carcinogenic potency (CP) of BaP and of the tumor growth rate (TGR) in TR-1 group animals, compared to the control and the TR-2 group. In marked contrast, complex 2 failed at the doses administered to exert any significant modulation of the above mentioned parameters. Results indicate that at low (100 micro/day) concentrations of vanadium, complex 1 exerts a significant anticarcinogenic effect on experimentally-induced leiomyosarcomas in rats, whereas complex 2 has no effect when administered at the same low concentrations of vanadium.

Beneficial effects of a vanadium complex with cysteine, administered at low doses on benzo(alpha)pyrene-induced leiomyosarcomas in Wistar rats.

BACKGROUND: Vanadium is a potent environmental and body metal, possessing remarkable antitumor and antidiabetic properties. Vanadium salts and complexes have been widely investigated for their anticarcinogenic properties in experimental carcinogenesis. In the present study the antitumor effects of a new vanadium complex with cysteine in relation to identical doses of vanadyl sulfate and cysteine, in tumor bearing rats are investigated. MATERIALS AND METHODS: Male wistar rats were injected with benzo(alpha)pyrene and divided into four groups of 21 rats each. Control group was treated only with BaP. The first group(TR-1) was treated by vanadyl sulfate per os at daily doses of 0.5 mg of V/kg b.w per day. The second (TR-2) by cysteine at doses of 4.5 mg/kg b.w per day and the third group (TR-3), by the complex V(III)-cysteine at daily doses of V 0.5 mg/kg b.w (containing cysteine at concentrations of 4.5 mg/b.w). Treatment was started when tumors were developed (evidenced from a palbable mass at the site of Bap injection) and went on till death. Toxicological tests were performed in 27 rats divided into a control group and two test groups; T-1 administered with vanadyl sulfate at daily doses of 18.5 mg V/kg b.w and T-2 group with V(III)-cysteine complex at daily doses of 18.5 V/kg b.w, for 9 weeks. Mean survival time, death rate, tumor growth rate, the carcinogenic potency of BaP, and the anticarcinogenic potency in relation to histological findings in each treatment group were calculated in each group in order to evaluate the antitumor effects of the substances used. RESULTS: Vanadyl sulfate, cysteine and V(III)-cysteine exerted antitumor effects on leiomyosarcoma bearing Wistar rats. However, V(III)-complex exerted much more potent effects than the other treatments, significantly prolonging mean survival time, retarding tumor growth rate and decreasing the carcinogenic potency of BaP in the TR-3 group, in comparison to the control and the TR-1 and TR-2 groups. Moreover V(III)-cysteine complex resulted in complete remission of 4 (19.7%) of the tumor bearing rats. Blood, urine, biochemical routine tests as well as autopsy did not reveal any toxic effects either of vanadyl sulafate or V(III)-cysteine complex. CONCLUSIONS: Vanadyl sulfate, cysteine and V(III)-cysteine complex exerted antitumor effects in tumor bearing rats. The V(III)-cysteine complex, however, exerts much more potent effects, as evident from the results of the present study. These beneficial effects of the above complex, in combination with its low toxicity provide evidence suggest its possible application in the treatment of human malignant diseases.

Antiproliferative and anticarcinogenic effects of an aqueous preparation of Abies alba and Viscum album se abies, on a L-1210 malignant cell line and tumor-bearing Wistar rats.

Extracts of plants have been widely tested for possible anticarcinogenic properties. In the present study a traditional remedy, consisting of an aqueous extract of mixed parts of the tree Abies alba and its mistletoe Viscum album se abies was tested on benzo(alpha)pyrene(BaP)-induced tumors in Wistar rats and on the L-1210 malignant cell line. Two main groups of male Wistar rats subcutaneously injected by 10 mg of BaP, a dose inducing 100% carcinogenesis, a control group (C-G, 15 rats) and a treatment group(TR-G, 18 rats), were used for the study. Five animals bearing BaP-induced tumors were also tested (TR-1-G). Animals of the TR-G were orally administered with the aqueous extract at doses of 50 ml/kg b.w, from the day of BaP injection and of the TR-1-G, from the 120th day of injection, till death. L-1210 malignant cells in cultivation, were administered with a powder obtained by condensation and lyophilization of the extract, at various concentrations and cytotoxicity was measured by the microculture tetrazolium assay. Autopsy of the rats, revealed metastasis in the lungs of the animals of all groups and the tumors developed were histologically identified as leiomyosarcomas. The results indicated that the extract of the above plants possess anticarcinogenic effects, documented by: a) its antiproliferative effects on L-1210 cells (IC50 = 49.6 +/- 1.4 micrograms/ml), b) the significant prolongation of life and reduction of tumor growth rate of the animals of the TR-G in comparison to the C-G, c) the inhibition by 16.6% of tumor induction in the TR-G and d) the prolongation of life and the necrotic effects of the extract on the tumors of the animals in the TR-1-G. The antiproliferative effects of the Abies alba and Viscum album se abies extract may be due to the lectins and thionins contained in Viscum album, as well as to the monoterpenes contained in Abies alba. Soft tissue tumors sensitive to the extract, are widespread among human organs, even in larynx, and are usually resistant to chemotherapy.

Is epinephrine-induced platelet aggregation autoregulated by its metabolic degradation products in vivo?

BACKGROUND: Catecholamines play an important role in platelet activation and aggregation, epinephrine being the most potent one. Catecholamines are substantially increased during stress, exercise or smoking and could result in clinically important platelet activation if their action was not rapidly regulated. In the present study the possible fast regulation of epinephrine-induced platelet aggregation by its metabolic degradation products is investigated. MATERIALS AND METHODS: Human platelet rich plasma (hPRP) and washed rabbit platelets(wRP) were used for the study. The platelets irreversible aggregation induced by epinephrine and ADP were monitored by an aggregometer prior to and after the addition of the catecholamines degradation products metanephrine, 3-methoxy-4-hydroxy-phenyl-glycole-aldehyde (MHPGA),3-methoxy-4-hydroxy-phenyl-mandelic acid (VMA) 3,4-dihydroxy phenyl glycole (DHPG),3,4-Dihydroxy-phenyl-glycole-aldehyde (DHPGA) and the trimethoxy-phenyl-methyl-piperazine(TMP), a known free radical scavenger and calcium antagonist. Linoleic acid-lipoxygenase reaction, in vitro was monitored in the presence and absence of VMA. RESULTS: Metabolic degradation products possessing a methoxy group at position 3 of the phenolic ring markedly inhibited epinephrine and ADP-induced platelet aggregation at microM concentrations. The most potent inhibitor of both agonists was metahephrine, followed by MHPGA and VMA. TMZ also inhibited platelet aggregation at concentrations similar to VMA. Dihydroxy-phenyl compounds failed to induce any inhibition. None of the substances tested induced any aggregatory effect even at high concentrations (1 mM). VMA significantly inhibited linoleic acid-lipoxygenase reaction at 0.1 microM. CONCLUSIONS: Results indicate that catecholamines' degradation products possessing methoxy (-OCH3) groups can rapidly inhibit in vitro and ex vivo epinephrine-induced platelet aggregation. The inhibitory effects of methoxy phenolic derivatives on epinephrine-induced platelet aggregation may possibly be attributed to their free radical scavenging properties. There is substantial evidence to conclude that an internal rapid autoregulation of epinephrine-induced platelet aggregation, caused by its metabolic degradation products, takes place in vivo.

Inhibition of benzo(a)pyrene-induced carcinogenesis by vitamin C alone and by vitamin C/vitamin E and selenium/glutathione.

The reduction on peroxidation caused by benzopyrenes by some naturally occurring antineoplastic agents was studied in this experimental work. Inhibition/reduction of experimental carcinogenesis induced by benzo(a)pyrene by vitamin C alone and by vitamin C/vitamin E and selenium/ glutathione was attempted in 224 female Wistar rats divided in four groups. Injected with 10.08 mL benzo(a)pyrene, the animals were treated with some naturally occurring substances like vitamin C alone and a combination of anticarcinogens. By calculating the carcinogenic potency of benzo(a)pyrene and the anticarcinogenic potency of substances used as well as histological examination of developed tumors and survival time of treated animals, it was found that vitamin C exerts a significant anticarcinogenic effect of 8.3 units and that the combination of the two anticarcinogens used produced a significant prolongation of the animals survival time with anticarcinogenic potency of 22.1 and 22.2 units, respectively. This is considered a potent anticarcinogenic effect. The question of an additional supportive administration of such agents complementary to the conventional cancer chemotherapy in humans is raised. Of course, further studies are needed.

The role of ascorbic acid, selenium, and glutathione on benzo[a]pyrene-induced carcinogenesis in wistar rats.

PURPOSE: The carcinogenic action of polycyclic aromatic hydrocarbons (PAHs) can be inhibited by endogenous or exogenous compounds. This study was designed to elucidate the modifying action of 3 endogenous inhibitors- ascorbic acid (vit C) used alone, and selenium (Se) used in combination with glutathione (GSH). MATERIALS AND METHODS: Chemical carcinogenesis was induced by benzo[a]pyrene(BaP). A hundred wistar rats were divided into 3 groups: the first group (G I) consisted of 42 animals, representing the control group. The two experimental groups (G II and G III) consisted of 38 and 20 rats, respectively. All groups were injected with BaP(10.08 mg subcutaneously-s.c). The first experimental G II was given only vit C (520 mg in 2% sugar solution per os - p.o.). The second experimental G III was given Se (0.1 mg p.o.) with GSH (200 mg p.o.). Tumor incidence and mean survival time were determined. Histological examination of the developed and excised tumors took place following death. The carcinogenic potency (CP) and anticarcinogenic potency (AP) of the substances used were calculated. RESULTS: A statistically significant difference regarding the mean survival time in the two experimental groups (238.4-/+31 days and 344.9-/+48 days, respectively) compared to the control group (183.8-/+28 days) was found (p < 0.001). The CP of each of the 3 groups was 54.3, 41.2, and 28.9 units, respectively. The AP of vit C used alone was 13.1 units, representing a significant anticarcinogenic effect. The combination of Se + GSH showed an AP of 25.4 units, resulting in a significant prolongation of the mean survival time, which is considered a potent anticarcinogenic effect. Furthermore, a statistically significant difference was found also when the mean survival time of G III animals was compared with G II. CONCLUSION: Vit C on its own and Se in combination with GSH represent strong endogenous inhibitors that can inhibit/reduce the carcinogenic action of BaP-induced carcinogenesis in wistar rats. The combination therapy used offered better in vivo results.

A simple procedure for estimation of total body surface area and determination of a new value of Meeh's constant in rats.

Precise calculation of total body surface area (TBSA) or premarked surface areas (P-MSAs) is of great importance in many biomedical applications. The aim of the paper was to present a simple procedure of measurement of P-MSAs in small animals and to determine a more accurate Meeh's constant (k), for a commonly used weight range of laboratory rats. A series of 30 Wistar rats, weighing 195-240 g, were anaesthetized and weighted. The TBSA of each animal was measured using a clear pocket and a planimeter. The data obtained were entered into the Meeh's formula (TBSA = kW(2/3)), the most commonly used for small experimental animals, so that a k value for each animal as well as a mean k value (9.83) were obtained. The TBSA of the animals was also calculated using the aforementioned mean k value and compared with that obtained using k values reported in previous studies. According to our findings, the new mean k value, determined with the use of our procedure of surface area measurement, ensured greater accuracy in the determination of the TBSA of experimental rats of a specific weight range. We also suggest a new procedure of surface area measurement which is easy, accurate and does not require animal sacrifice.

Intra-peritoneal application of catechins and EGCG as in vivo inhibitors of ozone-induced oxidative stress.

Oxidative stress is considered as a prominent feature of many acute and chronic diseases as well as of the normal aging process. We examined the effects of intra-peritoneal administration of catechins and EGCG as in vivo inhibitors of oxidative stress induced by ozone administration in two groups of Wistar rats. The first group was treated by intra-peritoneal administration of catechins and EGCG after the administration of ozone and the second group was pretreated by intra-peritoneal administration of catechins and EGCG prior to ozone administration. We determined in blood the activity of the enzymes superoxide dismutase and glutathione peroxidase, total antioxidant capacity, levels of copper and zinc and in urine malonaldehyde contents. Ozone administration resulted in significant reduction of glutathione peroxidase activity, plasma zinc levels and plasma and Red Blood Cells antioxidant capacity. Catechins and EGCG upregulate superoxide dismutase activity and maintain plasma and Red Blood Cells antioxidant capacity. Malonaldehyde levels at the end of the study were significantly increased only in the first group. Our data demonstrate that treatment with catechins and EGCG cannot reverse or prevent the effects of oxidative stress although some modulation occurs.

Anticarcinogenic and antiplatelet effects of carvacrol.

AIM: To investigate the effect of carvacrol on chemical carcinogenesis, cancer cell proliferation and platelet aggregation, and to find possible correlation between all these processes and the antioxidant properties of carvacrol. MATERIALS AND METHODS: 3,4-benzopyrene-induced carcinogenesis model using Wistar rats was used. Leiomyosarcoma cells from Wistar rats were used to study carvacrol antiproliferative activity in vitro. The carvacrol antiplatelet properties were investigated with platelet aggregation assay and flow cytometry technique. The production of thromboxane B2, final metabolite of platelet aggregation, was evaluated by radioimmunoassay. RESULTS: Our study revealed significant anticarcinogenic properties of carvacrol. We observed 30% decrease of 3,4 benzopyrene carcinogenic activity in vivo. Antiproliferative activity of carvacrol (IC(50)) was 90 microM and 67 microM for 24 h and 48 h of incubation of cells, respectively. Carvacrol possessed also mild antiplatelet effect, inducing the decrease of thromboxane A2 production in platelets and as a result - restrictive expression of the GPIIb/IIIa platelet receptor. CONCLUSION: Our data demonstrated that carvacrol possesses anticarcinogenic, antiproliferative and antiplatelet properties.

Effects of cigarette smoking on the antioxidant defence in young healthy male volunteers.

Cigarette smoking induces a significant oxidant effect related to variety of free radical-related diseases often affecting the upper respiratory tract, unless it is effectively compensated by the antioxidant barriers of the humans. In the present study, the evaluation of the antioxidant compensatory mechanisms, by estimating the antioxidant capacity of extracellular defence (saliva and plasma) and the intracellular resistance of peripheral lymphocytes to oxidative stress in young healthy smokers, was investigated. Twenty young healthy male smokers and 20 age-matched non-smokers with similar dietary profiles were enrolled in the study. Total saliva and plasma samples were collected from both groups, and total antioxidant capacity (TAC) and lag time were estimated. The latter was also repeated in smokers just after a cigarette smoking. Peripheral lymphocytes isolated from the subjects of both groups were also tested for their inherent DNA damage as well as for their ability to resist H2O2-induced DNA damage by using the comet assay. TAC of plasma was found significantly higher in smokers compared to non-smokers (p <0.004), whereas no difference was recorded in plasma lag time values. Lymphocytes of smokers manifested a significantly decreased oxidant resistance (increased DNA fragmentation) to H2O2, in comparison to non-smokers. Our results indicate that young smokers do not manifest different salivary antioxidant defence than non-smokers. They exhibit, however, a higher plasma antioxidant capacity, but a significantly reduced ability of blood lymphocytes, to resist to H2O2-induced DNA damage.

Trimetazidine for prevention of hepatic injury induced by ischaemia and reperfusion in rats.

OBJECTIVE: To assess the effect of trimetazidine (an anti-anginal drug that acts as a scavenger of oxygen free radicals) in the protection of hepatocytes after a 90 minute period of warm ischaemia followed by reperfusion in rats. DESIGN: Prospective study. MATERIAL: 80 Wistar rats. INTERVENTIONS: 20 Rats were given a single dose of trimetazidine 2.5 mg/kg intravenously 30 minutes before the induction of ischaemia; 20 received the same dose intraperitoneally twice a day for five days before the experiment and one dose intravenously 30 minutes before; 20 were given a single dose of 2.5 mg/kg intravenously after reperfusion had been started; and 20 acted as controls. All rats underwent liver biopsy through a laparotomy incision on postoperative days 2, 7, and 21, and the activities of liver enzymes in their blood were measured before induction of ischaemia and two, seven, 14, and 21 days afterwards. OUTCOME MEASURES: Histological changes and serum enzyme activities. RESULTS: The amount of centrilobular necrosis of hepatocytes, and the activity of hepatic enzymes were greatest on day 2, as was the reduction in superoxide dismutase activity in the erythrocytes. A single dose of trimetazidine, whether given before or after the ischaemic episode, gave significant protection to hepatocytes, but pretreatment for five days was even more effective. CONCLUSION: Trimetazidine protected rats' livers from injury after a period of warm ischaemia and reperfusion.

[Results of treatment of cystinuria and cystine lithiasis with alpha-mercaptopropionylglycine. Apropos of 40 patients]. / Résultats du traitement de la cystinurie et de la lithiase cystinique par l'alpha-mercaptopropionylglycine. A propos de 40 patients.

From 1987 to 1990, 719 subjects (385 males and 334 females) were screened, by means of urocystin-kit, for cystinuria and cystine lithiasis, in Epirus district (North-West Greece). The positive results were confirmed by chromatographic analysis of urine amino acids and laboratory tests. Forty patients, belonging to six cystinuric families, were identified. These patients were excreting 3.1 +/- 1.7 mmol/24 h of cystine in urine. All patients were treated by per os administration of alpha-MPG in daily doses of 400-1,200 mg/24 h). We therefore report the results of a 4 +/- 2 year follow-up of these patients. We conclude that treatment with alpha-MPG is very effective with minimal side effects in patients suffering from cystinuria or cystine urinary calculi.

Antitumour and toxic effects on Wistar rats of two new platinum complexes.

BACKGROUND: Cancer chemotherapy in humans based on metal complexes started at the clinical level in the late 1970s with the use of cisplatin, which forms intra-strand cross-links with DNA. METHODS: Two new platinum complexes of cis-geometry with the amino acids inosine (ino) and l-alanine (ala), Pt(ino)2Cl2 and cis-[Pt(NH3)2(ala)](NO3), respectively, were synthesized and pure samples were obtained by means of flash chromatography. These complexes were tested on benzo(a)pyrene-induced tumours in Wistar rats to detect their antitumour and toxic effects. RESULTS: There was a statistically significant prolongation of the mean survival time of the animals in the two groups tested (272 +/- 18 days and 246 +/- 26 days, respectively) compared to the control group (195 +/- 22 days) (P < 0.001). Toxic effects included a decrease in leucocyte cell count, mild haemolysis, mild haematuria, mild hepatotoxicity, elevated body temperature and hair loss. All of these effects were reversible after drug discontinuation. CONCLUSIONS: The two new platinum complexes described here appear to have an effective antitumour activity without severe toxicity when tested on Wistar rats.