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Epigallocatechin-3-gallate (EGCG) is widely used as a weight controlling supplement. Concerns about its safety evoked after cases of hepatotoxicity occurred upon its use. The underlying factors that could be involved in EGCG associated hepatotoxicity are not fully studied. In this study, we investigated the possible impact of lipopolysaccharide (LPS), as an inflammagen, on the effect of EGCG on hepatocytes. HepG2 cells were treated with different concentrations of EGCG (100, 200, 500 μM), with and without LPS (10 nM)-presensitization of the cells. Viability of HepG2 cells decreased with the increased concentrations of EGCG; the viability was even lesser in LPS-presensitized cells. Oxidative stress (Ox.LDL and CXCL16), the expression of nuclear retinoic receptors (RAR, RXR) and the biomarkers of hepatocellular injury (TNFα, TGFβ1) were all relatively higher in LPS-presensitized cells compared to non-sensitized cells upon treatment with EGCG. Sensitization of HepG2 cells with LPS alone did not affect the viability or any of the other biomarkers considered in this study. In conclusion, EGCG alone can be harmful to liver at high concentrations and this effect may become more pronounced under the influence of an inflammagen.

Since decades, it has been suggested that regular, moderate consumption of red wine, a major component of Mediterranean diet, at main meals, may contribute to explain the healthy properties attributed to this traditional dietary style. Despite preclinical in vitro/in vivo data have shown a significant cardioprotective activity of grape phytochemicals, mostly polyphenols, evidence in humans is still debated. This lack of consensus may be due to the equilibrium between the two main components of wine relevant for health: ethanol and bioactive compounds or phytochemicals, which include not only polyphenols, but also newly detected molecules, such as melatonin and phytosterols. The state of art related to this delicate equilibrium represents the starting point for designing future clinical trials, in perspective of clinical recommendations. A better comprehension of the wine chemistry complexity with its major components embodies a pivotal issue in biomedicine, involving the fields of diet-related environmental medicine as well as chronomedicine. In this paper, we briefly reviewed putative beneficial effects of moderate red wine intake in humans, focusing on the reduction of cardiovascular risk.

The prophylaxis against microbial airway infections of cystic fibrosis (CF) patients is an emerging application of chicken yolk antibody (IgY), however, no data on the effect of inhaled IgY have been published yet. Rats were daily (for 28 days) exposed to an aerosol of IgY, ovalbumin (OVA), Fab fragment of IgY, or PBS and their serum, bronchoalveolar lavage (BAL) and lung tissue were examined for inflammation signs. There were no marked changes in lung parenchyma, except for an elevated number of alveolar macrophages in the OVA-exposed group. While the administration of OVA or IgY aerosols slightly increased levels of cytokine TNF-α and GRO/KC in BAL fluid, a marked elevation of GM-CSF in serum was observed after the OVA inhalation. The administration of Fab induced expression of IL-1β > IL-18 in serum, in contrast no effect exerted by IgY. Our results suggest that the aerosolized IgY did not cause any deleterious effects in rat lungs.

Leptin down-regulates orexigenic and up-regulates anorexigenic neuropeptide gene expression in hypothalamus. Surgical removal of adipose tissue leads to decrease in circulating leptin concentrations in rats. In the present study, we tested: (a) regulation of neuropeptide gene expression in hypothalamus, (b) food intake, and (c) standard growth rate after removal of adipose tissue in rats. Partial lipectomy caused an approximately 10-fold reduction of subcutaneous, retroperitoneal and epididymal adipose tissue weight (at the end of experiments adipose tissue weight was 1.5 ± 0.9 in lipectomy and 15 ± 3.9 g in control rats; statistically significant). Compared to control rats, the animals subjected to lipectomy presented increased food intake, standard growth rate, and decreased serum leptin concentrations (2.6 ± 0.8 vs. 3.7 ± 1.2 ng/mL in the controls, statistically significant). These changes were associated with approximately twofold increase in neuropeptide Y, threefold increase in agouti-related peptide (orexigenic neuropeptides) and about 50% decrease in pro-opiomelanocortin and cocaine-amphetamine-regulated transcript peptide (anorexigenic neuropeptides) mRNA levels in the hypothalamus. These results suggest that partial lipectomy, leading to a decrease in circulating leptin concentrations, may exert an effect on hypothalamic orexigenic and anorexigenic neuropeptide gene expression, and consequently modulate food intake and standard growth rate in rats.

The genus Bothrops is responsible for approximately 90% of snakebites in Brazil. In the present study biomarkers of oxidative stress (OS) were evaluated in the blood of victims of snakebites from Bothrops jararaca and Bothrops jararacussu. Patient monitoring started from the emergency entrance at the hospital up to 30 days, groups divided as follows: time 0 (t₀), 24 hours (t_24h), 7 days (t_7d) and 30 days (t_30d). The activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), and myeloperoxidase (MPO), as well as the contents of reduced glutathione (GSH), vitamin E, lipid peroxidation (TBARS), protein carbonyls (PC) were examined in blood. Initial determinations revealed increased CAT, GR and GPx activities and decreased SOD and GST activities together with the depletion of GSH contents, while markers of oxidative damage showed increased TBARS levels and decreased PC concentrations in victims of snakebite compared to controls (blood donors). Regarding the temporal effect, no statistical differences among the groups were detected for the distinct parameters analyzed. The responses obtained in OS biomarkers in victims of snakebite compared to healthy subjects indicate that Bothrops envenomation promoted a pronounced and persistent systemic OS in the blood of those subjects.

Large screening programs of nasopharyngeal carcinoma (NPC) are still challenging due to the scarcity of the resources in higher prevalence parts of the world. In the present study, the development of a new low cost immunochromatography (ICG) strip test for screening of nasopharyngeal carcinoma is reported. A conjugate of gold nanoparticle-anti viral capsid antigen (VCA) immunoglobulin A (IgA) antibody was used as the detection probe. The test is based on a non-competitive immunoassay for rapid and reliable detection the serum specific IgA of Epstein-Barr virus (EBV) VCA. Patient serum samples were analyzed with the strip and compared with enzyme-linked immunosorbent assay (ELISA) and pathological evaluation. The obtained results from the ICG strips were well correlated with the two reference methods. An evaluation of stability, sensitivity and specificity suggested the strip suitable for NPC screening programs in high-risk peoples and also for the recurrence monitoring.

Silibinin is known to display high efficacy against cancer cells and for hepatic protection. Metformin, a well-known antidiabetic agent, has recently been reported to inhibit cancer. In the present study, we investigated the effect of metformin on silibinin-induced programmed cell death in cervical cancer cells (C-33A). MTT assay and Western blot assays were performed to quantify cell viability and the expression of signaling proteins, respectively. Combined treatment with metformin and silibinin decreased cell survival in synergistic manner in C-33A cells at a dose that did not affect nonmalignant cells (HUVECs). Silibinin and metformin increased PTEN and AMPK expression in C-33A cells, respectively. Combined treatment caused a greater increase in the expression of activated caspase-3 or AIF, indicating apoptosis. Combined treatment with silibinin and metformin may induce programmed cell death of human cervical cancer cells at a dose that does not affect HUVECs. This finding reveals a potential therapeutic strategy of cervical cancer.

Electrogastrography (EGG) is a non-invasive method for the assessment of gastric myoelectrical activity. Porcine EGG is comparable with human one. There are no data on the impact of moderate to high doses of atropine on EGG, neither in humans nor experimental pigs. The purpose of this study was to evaluate the effect of different doses of atropine on EGG in experimental pigs.
Six fasting pigs entered the study three times in a random order. The baseline EGG recording lasted 20 min, followed by a 105-min EGG trial recording. Intramuscular atropine 1.5 mg (part 1), 3.0 mg (part 2) and 4.5 mg (part 3) was administrated after the baseline EGG.
Atropine doses of 1.5 and 3.0 mg revealed a similar pattern in the EGG power course. After an initial increase (at the first 15-min interval), the areas of amplitudes decreased back to values comparable with basal levels and subsequently increased significantly again to the maximum at 105 min. The EGG power was quite different after the administration of 4.5 mg of atropine. Areas of amplitudes decreased gradually to the minimum values at 105 min after atropine administration.
In conclusion, different dose-dependent changes in the EGG pattern were found after moderate to high doses of atropine.

MSCs produce CD4(+)CD25(+)FOX3(+) regulatory T (Treg) cells from activated peripheral blood mononuclear cells (PBMC), T-CD4+ and T-CD8+ cells in vitro and in vivo. Here we investigated whether the deficiency of androgen/AR in MSCs influence Treg induction from total PBMC, splenocytes, CD4+CD25-through AR/TGF-β interaction. Eight to 12-week-old wild type and general androgen receptor knockout (ARKO) mice were used. MSCs were collected, characterized and function of Treg cells was studied. Our result showed that depletion of AR suppressed the immunosuppressive effect of MSCs, and demonstrated that WT-MSC-induced Treg cell expansion was partially impaired by blocking androgen receptor signal. Furthermore, the levels of TGF-β were lower in the T cell coculture with ARKO-MSC compared to WT-MSC. Exposure of ARKO-MSC cells to exogenous active TGF-β partially restored the induction of Treg cell expansion by ARKO-MSC cells. Our data suggest that ARKO-MSC hampers Treg cell expansion and function via androgen/AR and TGF-β signal pathways interaction. To the best of our knowledge, this study is the first investigating the interaction of MSCs from ARKO mice and WT Tregs in an allogeneic co-culture model. Together, these results might provide great insight into treatment of inflammatory and autoimmune diseases.

Citrulline ureidase (CTU, EC3.5.1.20) degrades citrulline into ornithine, carbon dioxide, and ammonia. Here, we present the report on expression of recombinant CTU in Escherichia coli. The soluble and active recombinant CTU was expressed in the periplasmic space with the vector pET-22b and the His-tagged CTU was purified with Ni-Affinity Chromatography. The yield of soluble recombinant protein was significantly increased when 1% sorbitol was supplemented in medium. By using phenylisothiocyanate (PITC) pre-column derivatization HPLC, the enzyme activity of recombinant CTU was determined via measuring of the substrate citrulline and the corresponding products. Our results could be useful in the study of CTU biochemical characteristics, enzymatic preparation of ornithine and development of an enzymatic detection method of citrulline.

The present work aimed at purifying the intracellular fungal metabolites, such as 16-methylheptadecanoic acid methyl ester (HDA) and 9,12-octadecadienoic acid (ODA) from marine Trichoderma, Hypocrea lixii TSK8, Hypocrea rufa SKS2 respectively, and investigating their anticancer and antioxidant effects. The two fungal metabolites were tested against two human cancer cell lines, namely oral cancer (KB) and skin carcinoma (A431) by using MTT assay. The inhibitory concentrations (IC₅₀) against KB oral cancer cells were found to be 18.75 ± 0.12 μg/mL for HDA and 75.50 ± 0.42 μg/mL for ODA. Whereas IC₅₀ values of HDA and ODA against A431 were found 37.5 ± 0.42 μg/mL and 72.89 ± 0.15 μg/mL, respectively. In addition, the down-regulation of heat shock protein 90 kDa (HSP90) was confirmed by using SDS-PAGE and Western blot analysis. The effect of HDA induced apoptosis via ROS-dependent internucleosomal DNA fragmentation was confirmed by AGE analysis. We further evaluated the in vivo anti-skin cancer activity of HDA in Swiss albino mice induced with skin cancer by 7,12-dimethylbenz(a)anthracene (DMBA) and croton oil (CO). The in vivo hematological, biochemical and histopathological results revealed that the fungal metabolite HDA was a highly potent anticancer compound against the skin cancer.

Stereospecific nucleation of mesoporous hybrid microspheres composed of CaCO₃ and carrageenan was appended to one side of bacterial cellulose membrane synthesized in static cultures of Gluconacetobacter hansenii to develop an implantable drug delivery device. The synthesis of the hybrid microparticles proceeds by self-assembly mechanism in the presence of calcium and contains tailorable amounts of doxorubicin. However, in the absence of the particles, doxorubicin was distributed along the BC film, but without control release of drug. Infrared spectroscopy, confocal and scanning electron microscopies analyses demonstrate that the doxorubicin is entrapped inside the hybrid particles with approximately 80% drug loading compared to the 11% obtained for native bacterial cellulose. Doxorubicin content in the hybrid particles can be increased by a factor of 10 (from 258.6 to 2586.3 nmol ml^-1), and also by the quantities of particles regulated by the CaCO₃-carrageenan content and the physicochemical microenvironment. The hybrid BC system can be considered as smart device since the kinetic release of doxorubicin from the hybrid cellulose system rise from 1.50 to 2.75 μg/membrane/day when the pH decreases from 7.4 to 5.8 at 37 °C, a pathologic simulated environment. The hybrid microparticle system can be potentially used as an implantable drug delivery system for personalized oncological therapies.

This study investigated the effects of sericin-derived oligopeptides on natural killer (NK) activity. In vitro exposure of human peripheral blood mononuclear cells with sericin-derived oligopeptides resulted in an augmentation of NK cell activity against K562 target cells and the effects appeared to be dose-related. Experiments designed to examine whether enhanced NK activity was due to direct or indirect activation of NK cells revealed that sericin oligopeptides did not induce activity of purified NK cells, and that sericin oligopeptides augmented NK activity indirectly by inducing the production of IL-2 and IFN-γ cytokines. In in vivo experimentation where mice were orally administered with sericin oligopeptides and splenic mononuclear cells tested against YAC-1 target cells, significant increase in NK activity was obtained compared to control mice. Elevated levels of IL-2 were also evident in all oligopeptides-treated groups. As demonstrated both in vitro and in vivo, these results indicate that sericin-derived oligopeptides have efficient NK-enhancing activity and suggest the potential therapeutic applications of such oligopeptides for functional improvement of NK cells, and possibly for treatment of tumor and infectious diseases in which NK activity contributes to host defense.

The aim of this study was to evaluate the effectiveness of individual (inactive) proenzymes and mixtures thereof in cancer treatment and to compare this treatment with more frequently used therapy based on active proteases. Experiments focused on explanation of possible mechanisms of proenzyme action against tumours are included.
Proenzyme therapy of sarcoma S-180 significantly reduced tumour growth and prolonged survival of mice. The effect of trypsinogen and chymotrypsinogen was synergistic. Proenzyme therapy of melanoma B16-F10 bearing mice reduced both tumour growth and prevalence of metastases. Active enzyme based therapy of melanoma B16-F10 was less effective. Severe combined immunodeficiency (SCID) mice bearing sarcoma S-180 did not respond to the proenzyme therapy, indicating that the effect of this therapy is dependent on fully developed acquired immunity. Measured decreased levels of TGF-β and an increased amount of alpha-2 macroglobulin in serum contributed to the elucidation of the cancer treatment mechanism.
Proenzyme therapy based on administration of a mixture of trypsinogen and chymotrypsinogen is effective in cancer treatment.

The possibility of regeneration in a damaged central nervous system (CNS) is limited and therapy of CNS diseases accompanied with neuronal loss is problematic.
Lurcher (Lc) mutant mice represent a model for olivocerebellar degeneration.
Heterozygotes suffer from a complete loss of cerebellar Purkinje cells and a secondary reduction of granule cells and inferior olive neuron number.

Elephantopus scaber Linn., family Asteraceae, is a small herb found in Neotropics, Europe, Asia, Africa and Australia. The plant parts of this herb have been used traditionally for the treatment of a number of diseases in many countries. Sesquiterpene lactones, triterpenoids, steroids, flavonoids and essential oil constituents have been reported from various parts of the plant. The plant has been extensively screened for anticancer activity. Sesquiterpene lactones such as deoxyelephantopin, isodeoxyelephantopin, scabertopin, and isoscabertopin have been found to be prominent anticancer constituents. Many other biological activities such as antimicrobial, hepatoprotective, antioxidant, antidiabetic, anti-inflammatory, analgesic, antiasthamatic, antiplatelet, and wound healing have been reported in various research papers. The present review has been envisaged with an intension to provide scientific information about the ethnomedical, phytochemical and pharmacological profile of E. scaber.

The neuroprotective effects of a novel oxime KR-22934, the oxime K203 and obidoxime in combination with atropine in rats poisoned with tabun at a sublethal dose (200 μg/kg i.m.; 80% LD₅₀) were studied. The tabun-induced neurotoxicity was monitored at 24 h following tabun challenge using a functional observational battery and an automatic measurement of motor activity. The results indicate that all tabun-poisoned rats treated with oximes in combination with atropine were able to survive within 24 h following tabun poisoning. One tabun-poisoned rat without antidotal treatment died within 24 h. The oximes KR-22934 and K203 combined with atropine showed a similar potency to decrease tabun-induced neurotoxicity at 24 h after tabun administration while the neuroprotective efficacy of obidoxime was slightly higher. However, no oxime was able to eliminate tabun-induced neurotoxicity completely. When atropine was administered alone, negligible neuroprotective efficacy was observed. Based on the results, a novel oxime KR-22934 did not bring any improvement of the neuroprotective efficacy of antidotal treatment of acute tabun poisonings.

Epidemiological data indicate that selenium status is inversely connected with cancer risk. Animal and human studies have demonstrated that most inorganic and organic forms of selenium compounds have an anticancer action. This work investigated the impact of organic selenium on the multiple signalling pathways involved in the inhibition of the viability of prostate cancer cells. Prostate adenocarcinoma cells (PC-3) were incubated with seleno-l-methionine (SeMet) at four concentrations and cell viability and programmed cell death were determined by the WST-1, BrdU assays and Tali image based cytometer. The expression of chosen cell-cycle regulatory genes was determined by real-time RT-PCR analysis and confirmed at the protein level. SeMet treatment of PC-3 cells resulted in an inhibition of cell proliferation in a dose- and time-dependent manner. The inhibition of proliferation correlated with the up-regulation of gene expression and the protein levels of CCNG1, CHEK1, CDKN1C and GADD45A, whereas SeMet down-regulated the expression of CCNA1 and CDK6 genes. Therefore SeMet inhibits the proliferative activity of prostate cancer cells by a direct influence on the expression of genes involved in the regulation of cell cycle progression.

PPARδ-dependent maintenance of inotropic function is mentioned as crucial for cardiomyocytes. However, change of PPARδ in endotoxins-induced cardiac dysfunction is still unclear. The present study is then designed to investigate the changes of PPARδ in rats showing LPS-induced cardiac dysfunction. In the in vivo experiments, adult Wistar rats were treated with intravenous injection of 10 mg/kg LPS for 6 h. The isolated heart determined in Langendorff apparatus and the hemodynamic analysis of rats used to measure the changes of cardiac function extra vivo and in vivo. We found that LPS decreased the cardiac contractility in isolated heart and lowered the hemodynamic dP/dt_max in rats. Also, this action of LPS was reversed by PPARδ agonist. In cultured neonatal rat cardiac cells incubated with LPS, the intracellular calcium concentration and troponin I phosphorylation were both reduced after the detection of intracellular calcium level and Western blotting analysis. PPARδ agonist also reversed both actions of LPS in cardiomyocyte. The obtained results suggest that LPS induced decreases in PPARδ expression and troponin I phosphorylation to result in acute heart failure similar to cardiac dysfunction in endotoxemia.

This work aims at studying in children the prevalence of some vascular variability anomalies (VVAs), i.e. indices of an increased risk of heart and vessel diseases revealed by chronobiologic analysis of cardiovascular monitoring data - systolic (SBP) and diastolic (DBP) MESOR-hypertension/hypotension, circadian hyper-amplitude-tension (CHAT), ecphasia and an excessive pulse pressure (EPP). A reference group of 138 healthy Moscow schoolchildren (age 12-17 years) was used for determination of 90% prediction limits for the circadian parameters of SBP and DBP, specified by gender and age (or height). The test group included another 194 children of the same age range with headache and abnormal BP values by casual measurement. BP data series from each child were analyzed by the COSINOR method. VVAs were found in 44.8% of children from the test group. In boys, VVAs occur more often than in girls (51.5% vs. 30.6%). One child may have up to 3 VVAs. The presence of VVAs may be related to an elevated BMI. The most common VVAs in childhood are the abnormalities of PP and systolic CHAT. Cosinor analysis can reveal a large number of children with PP higher than a threshold (60 mmHg) during a relatively long portion of the day (more than 1/3 of the 24 h span).

Recombinant human bone morphogenetic protein-2 (rhBMP-2) is an osteo-inductive reagent that can be used therapeutically to promote spinal arthrodesis and fracture healing. Despite its clinical use, the effect of rhBMP-2 on expression of endogenous osteogenic growth factors is poorly described. The goal of this study was to determine if local rhBMP-2 treatment modulates expression of pro-osteogenic growth factors during fracture healing. Rat femur fractures were wrapped with control or rhBMP-2 collagen soaked sponge. Radiographic observations demonstrated that rhBMP-2 treatment enhanced fracture callus formation and decreased bridging time. Expression of fracture site growth factor mRNAs was determined by quantitative polymerase chain reaction. The gene expression data demonstrated that rhBMP-2 treatment significantly increased mRNA levels of BMP-2 at day 4, BMP-4 at days 2, 4, 7, and 10, BMP-7 at day 2, and TNF-α at day 7. Treatment with rhBMP-2 was shown to significantly reduce mRNA levels of BMP-2 on day 2 and 10, TNF-α on day 14, and BMP-6 on days 2, 10, 14, and 21. TGFβ-1 was also significantly reduced on days 2 and 21. In conclusion, treatment with rhBMP-2 appears to enhance expression of other osteogenic factors that could contribute to bone formation.

Antibacterial activity of silver nanoparticles is widely known and used. The effect of application of therapeutical ultrasonic field in the presence of silver nanoparticles

Cholinesterase inhibitors are beneficial in the treatment of Alzheimer's disease via indirect increase of cholinergic neuro-transmission. The aim of the present study was to evaluate the potency of inhibitors tacrine, rivastigmine and donepezil to reverse cholinergic depletion induced by 3-quinuclidinyl benzilate (QNB, 2 mg kg^-1) in Wistar rats performing the multiple T-maze test. The effect of QNB on retention was compared to the effect of standard amnesic drug, scopolamine, at the dose of 0.3 mg kg^-1. Well-trained rats were treated intra-peritoneally with QNB, followed by another injection containing saline or tacrine (10 mg kg^-1) or rivastigmine (1.2 mg kg^-1) or donepezil (2.65 mg kg^-1) 15 min later. Rats were subjected to the T-maze task 30 min and 24 h following QNB administration. The passage time and number of errors were observed. QNB significantly impaired the performance of rats in both tested times in contrast to short-lasting effect of scopolamine (30 min only). The inhibitors rivastigmine and donepezil significantly attenuated QNB-induced behavioural impairment in the 30 min tests, whereas tacrine failed to have the same effect. Moreover, the performance of tacrine-treated rats was worse due to cholinergic over-stimulation. Beneficial effects of all tested inhibitors including tacrine were evident in the 24 h test.

Quaternary ammonium salts (chloride, bromide and iodide; QUATs) with n-alkyl chain lengths between C8 and C18 have been synthesized under optimized experimental conditions. These compounds were tested in vitro for antimicrobial activity against representative bacterial strains (Staphylococcus aureus CIP 4.83, Enterococcus hirae CIP 5855, Pseudomonas aeruginosa CIP 82118, Escherichia coli CIP 53126, Mycobacterium smegmatis CIP 7326) and fungal species (Aspergillus niger ATCC 16404, Candida albicans IP 118079, Trichophyton interdigitale IP 146583). While these compounds showed moderate antifungal activity, several of them (particularly C14-I^-) may be considered as highly potential antibacterial agents against S. aureus, E. hirae and E. coli with MIC values lower than that of commercial benzalkonium chloride and ciprofloxacin used as standards. The relationship between the lipophilicity and the antibacterial activity of the tested QUATs was quantified by a multiple linear regression method.

Platelets plays a central role in hemostatic processes and consequently are similarly involved in pathological processes, such as arterial thrombosis and atherosclerosis. In this study we investigate the effect of aqueous crude extracts of Scutia buxifolia on NTPDase and 5'-nucleotidase activity on platelets and lymphocytes as well as the profile of the platelet aggregation. The effect of the aqueous crude extract obtained from S. buxifolia leaves (SbL) and stem bark (SbS) on enzymatic activities and platelet aggregation was investigated by in vitro tests. The platelets and lymphocytes were exposed to aqueous extracts of S. buxifolia at concentrations 1-200 μg/mL in the presence of ATP, ADP, AMP as substrates. The results showed that SbS and SbL potentially inhibited the NTPDase and 5'-nucleotidase in platelets and lymphocytes. Moreover, we found that ADP-induced aggregation was only inhibited by the SbS. This data suggest the existence of compounds in S. buxifolia that may decrease the NTPDase and 5'-nucleotidase activity, causing peripheral alterations in adenine nucleotide levels and protection against ADP-induced platelet aggregation.

The aim of the study was to investigate clock gene expression in Bos taurus and the alteration of that during two pathological conditions, evaluating the daily expression pattern of four clock genes (Per2, Cry2, Bmal1, Clock) in peripheral blood cells. Five healthy cows, five affected by Brucellosis (BR) and five affected by Bovine Viral Diarrhoea-Mucosal Disease (BVD-MD) were housed in indoor stalls under natural spring conditions, blood samples were collected at 4 h intervals over a 24 h period. Statistical analysis showed rhythmic expression of clock genes mRNAs in healthy cows. Cows affected by BR did not show any rhythmic expression of clock genes mRNAs, cows affected by BDV mRNA levels of Bmal1, Clock and Cry2 changed during the day. These findings highlighted that circadian system could be involved in homeostasis alteration and that clock genes could be considerate as regulatory genes or early response genes during inflammation, so, their regulation should be evaluated in health research and treatment.

Diethyldithiocarbamate (ditiocarb), a metabolite of the old anti-alcoholic drug disulfiram (Antabuse), forms proteasome-inhibiting metal complexes with copper or zinc that suppress cancer cells both in vitro and in vivo. The drug has been used in a clinical trial (NCT00742911) along with copper gluconate as a dietary supplement in patients with cancer spreading to the liver. In this study, we demonstrate the effect of synthetic complexes of disulfiram with four various metals (Mn, Fe, Cr and Cu) used as food supplements. These complexes may be spontaneously formed in the blood during the use of disulfiram with divalent metals and thus may suppress the growth of cancer in vivo. The cytotoxic effect of the compounds and the compounds' ability to inhibit the cellular proteasome were tested in the osteosarcoma cell line U2OS. After 48 h, copper and manganese complexes exhibited cytotoxic effect on the cell line, in sharp contrast to both iron and chromium complexes.

New components of transdisciplinary spectra or known components in new variables in us, matching those around us, are being mapped. Their hardly trivial interactions associated with the good and bad around us - from religiosity to crime and war - are being rendered measurable, for the eventual development of countermeasures to the diseases of societies and nations. Internal cycles not only underlie life itself and underlie our evolving genetics at all levels of organization; they also constitute the essential control and reference information in all transdisciplinary science. In preparing for travel to Mars and other missions in space that may take more than a year, let us do what is immediately practicable. Transyears may have very small amplitudes yet are associated with sudden cardiac death in some terrestrial locations; if they should play a role in these electrical incidents of the heart, among others like myocardial infarction and stroke, they will jeopardize lengthy missions in extraterrestrial space, away from hospitals.
The likelihood of stroke or cardiac death can be immediately reduced by chronobiologically assessing blood pressure and heart rate variability and by optimizing the efficacy of timed treatment rather than relying on an unacceptable and often inaccurate spotcheck and treating by convenience rather than pertinence. Needed are: detection of nocturnal abnormality when medication may no longer be effective (or is too effective) neither seen during office visits by day; detection of circadian hyper-amplitude-tension (CHAT) associated with a risk of stroke and kidney disease greater than other risks (including "hypertension" when all risks are assessed concomitantly); detection of CHAT as high risk among normotensives who may not need anti-hypertensive medication; individualized inferential statistical testing to determine whether a drug or non-drug intervention such as autogenic training (relaxation) is effective and for how long (detecting any initial and later success or failure), some of which conditions otherwise are not found without chronobiology; individualization of treatment timing, since the same dose of the same medication can further lower the subject's blood pressure average and circadian amplitude when the timing of daily administration is optimized, as ascertained by sequential testing and parameter tests.
Thus, we save lives by monitoring and assessing, and if need be treating, vascular disease risk through chronobiologically interpreted 24-hour or preferably longer (24-hour/7-day) blood pressure and heart rate variability. Abnormalities in the variability of blood pressure and heart rate, impossible to find during a conventional office visit (the latter aiming at the fiction of a "true" blood pressure), can raise cardiovascular disease risk in the next six years from 4% to 100%.

Apigenin is a member of the flavone family of flavonoids and possesses anti-inflammatory, free radical scavenging and anti-carcinogenic properties. Hydrogen peroxide, which is generated during oxidative stress, is known to damage proteins, nucleic acids and cell membranes and also has been implicated in cancer, ageing and several chronic neurogenerative diseases. The present study focuses on the protective effect of apigenin against genotoxic doses of hydrogen peroxide (H₂O₂) using sister chromatid exchanges (SCEs) and cytokinesis blocked micronucleus (CBMN) assay. The treatment with 50, 100 and 150 μM of H₂O₂ results in a significant dose dependent increase in the frequency of SCEs and MN. The treatment with 100 μM of H₂O₂ along with 5, 10 and 20 μM of apigenin results in a dose dependent significant decrease in the frequency of SCEs and MN on cultured human lymphocytes. A similar result was obtained with treatment with 150 μM of H₂O₂ along with 5, 10 and 20 μM of apigenin. The results of the present study suggest a protective effect of apigenin against hydrogen peroxide induced genotoxic damage on cultured human lymphocytes.