Cornelian Cherry (Cornus mas L.) Extracts Exert Cytotoxicity in Two Selected Melanoma Cell Lines-A Factorial Analysis of Time-Dependent Alterations in Values Obtained with SRB and MTT Assays.

Despite the fact that phytochemicals of Cornaceae species have long been discussed as possible auxiliary agents in contemporary treatment, the insights on their properties remain relatively scarce. This study focuses on Cornus mas L. (Cornelian cherry), the extracts of which are reported to exert a pleiotropic effect shown in both in vivo and in vitro studies. This study aimed to explore the cytotoxic effect of extracts from fruits of red (Cornus mas L. 'Podolski') and yellow (Cornus mas L. 'Yantarnyi' and 'Flava') Cornelian cherries on two melanoma cell lines (A375 and MeWo). The extracts were characterized in the context of the concentration of bioactive compounds of antioxidative properties. Cytotoxicity was investigated with the use of the following two assays: SRB and MTT. An additional, alternative protocol for the SRB assay was used in this study so as to account for possible bias. Cytotoxicity was assessed as a difference in the whole time series of cell viability, instead of analyzing differences in raw values (often found in the literature). Both extracts from Cornus mas L. induced cytotoxicity in both A375 and MeWo cell lines, although the response of these cells was different. Moreover, based on this study, there is no evidence for claiming a different magnitude of cytotoxicity between these two extracts.

L-Arginine/Nitric Oxide Pathway Is Altered in Colorectal Cancer and Can Be Modulated by Novel Derivatives from Oxicam Class of Non-Steroidal Anti-Inflammatory Drugs.

L-arginine/nitric oxide pathway metabolites are altered in colorectal cancer (CRC). We evaluated underlying changes in pathway enzymes in 55 paired tumor/tumor-adjacent samples and 20 normal mucosa using quantitative-PCR and assessed the impact of classic and novel oxicam analogues on enzyme expression and intracellular metabolite concentration (LC-MS/MS) in Caco-2, HCT116, and HT-29 cells. Compared to normal mucosa, ARG1, PRMT1, and PRMT5 were overexpressed in both tumor and tumor-adjacent tissue and DDAH2 solely in tumor-adjacent tissue. Tumor-adjacent tissue had higher expression of ARG1, DDAH1, and DDAH2 and lower NOS2 than patients-matched tumors. The ARG1 expression in tumors increased along with tumor grade and reflected lymph node involvement. Novel oxicam analogues with arylpiperazine moiety at the thiazine ring were more effective in downregulating DDAHs and PRMTs and upregulating ARG2 than piroxicam and meloxicam. An analogue distinguished by propylene linker between thiazine's and piperazine's nitrogen atoms and containing two fluorine substituents was the strongest inhibitor of DDAHs and PRMTs expression, while an analogue containing propylene linker but no fluorine substituents was the strongest inhibitor of ARG2 expression. Metabolic reprogramming in CRC includes overexpression of DDAHs and PRMTs in addition to ARG1 and NOS2 and is not restricted to tumor tissue but can be modulated by novel oxicam analogues.

L-Arginine/NO Pathway Metabolites in Colorectal Cancer: Relevance as Disease Biomarkers and Predictors of Adverse Clinical Outcomes Following Surgery.

The L-Arginine/NO pathway is involved in carcinogenesis and immunity. Its diagnostic and prognostic value in colorectal cancer (CRC) was determined using tandem mass spectrometry in 199 individuals (137 with CRC) and, during a three-day follow up, in 60 patients undergoing colorectal surgery. Citrulline was decreased and asymmetric (ADMA) and symmetric (SDMA) dimethylarginines and dimethylamine (DMA) were increased in CRC. The DMA increase corresponded with CRC advancement while arginine, ADMA, and SDMA levels were higher in left-sided cancers. Arginine, citrulline, ADMA, and DMA dropped and SDMA increased post incision. Females experienced a more substantial drop in arginine. The arginine and ADMA dynamics depended on blood loss. The initial SDMA increase was higher in patients requiring transfusions. Postoperative dynamics in arginine and dimethylarginines differed in robot-assisted and open surgery. Concomitant SDMA, citrulline, and DMA quantification displayed a 92% accuracy in detecting CRC. Monitoring changes in arginine, ADMA, and SDMA in the early postoperative period predicted postoperative ileus with 84% and surgical site infections with 90% accuracy. Changes in ADMA predicted operative morbidity with 90% and anastomotic leakage with 77% accuracy. If positively validated, L-arginine/NO pathway metabolites may facilitate CRC screening and surveillance, support differential diagnosis, and assist in clinical decision-making regarding patients recovering from colorectal surgery.

Interleukins 4 and 13 and Their Receptors Are Differently Expressed in Gastrointestinal Tract Cancers, Depending on the Anatomical Site and Disease Advancement, and Improve Colon Cancer Cell Viability and Motility.

Immunosuppressive interleukins (IL)-4 and 13 may directly promote cancer but neither their status nor role in gastrointestinal tract is clarified. We aim at quantifying ILs and their receptors in paired normal-tumor samples (n = 49/51) and sera (n = 263), using immunoassays and RTqPCR, and screening for their effect on colonic cancer cells. Both ILs were elevated locally at protein level in all cancers but only IL13 transcripts in colon were upregulated. Interleukin and their receptor expression reflected cancer pathology to varying degrees, with the association frequently inverse and manifested in non-cancerous tissue. Positive correlation with cancer-promoting genes BCL2, BCLxL, HIF1A, VEGFA, ACTA2, CCL2, PTGS2, and CDKN1A, but not Ki67, was demonstrated, particularly for ILs' receptors. Circulating IL-4 was elevated in all, while IL-13 only in colorectal or esophageal cancers, reflecting their advancement. IL4Ra and IL13Ra1 transcripts were downregulated by hypoxia and, in Caco-2, also by IL-4. Interleukin stimulation slightly improved colonic cancer cell viability, weakly upregulating BCL2 and Ki67 in HCT116 and HT-29. It affected cell motility more markedly and was consistently accompanied by upregulation of claudin-2. Gastrointestinal tract cancers are associated with IL-4 and IL-13 upregulation, which may facilitate cancer growth. Targeting both interleukins as an antineoplastic strategy warrants further investigation.

Left extrastriate body area is sensitive to the meaning of symbolic gesture: evidence from fMRI repetition suppression.

Functional magnetic resonance imaging (fMRI) adaptation (a.k.a. repetition suppression) paradigm was used to test if semantic information contained in object-related (transitive) pantomimes and communicative (intransitive) gestures is represented differently in the occipito-temporal cortex. Participants watched 2.75 s back-to-back videos where the meaning of gesture was either repeated or changed. The just observed (typically second) gesture was then imitated. To maintain participants' attention, some trials contained a single video. fMRI adaptation -signal decreases- for watching both movement categories were observed particularly in the lateral occipital cortex, including the extrastriate body area (EBA). Yet, intransitive (vs. transitive) gesture specific repetition suppression was found mainly in the left rostral EBA and caudal middle temporal gyrus- the rEBA/cMTG complex. Repetition enhancement (signal increase) was revealed in the precuneus. While the whole brain and region-of-interest analyses indicate that the precuneus is involved only in visuospatial action processing for later imitation, the common EBA repetition suppression discloses sensitivity to the meaning of symbolic gesture, namely the "semantic what" of actions. Moreover, the rEBA/cMTG suppression reveals greater selectivity for conventionalized communicative gesture. Thus, fMRI adaptation shows higher-order functions of EBA, its role in the semantic network, and indicates that its functional repertoire is wider than previously thought.

The efficacy of an education-based secondary outpatient prevention programme after acute coronary syndrome hospitalisations and treatment in Poland. The Patient Club initiative.

BACKGROUND: Guidelines on cardiovascular prevention from the European Society of Cardiology advocate sustained educational measures to be undertaken by both doctors and nurses to ensure lifestyle changes for patients after acute coronary syndromes (ACS). A planned programme of a series of educational meetings for patients after ACS was initiated in form of the Patient Club. AIM: To assess the efficacy of the Patient Club initiative in terms of increased knowledge on cardiovascular risk factors as well as utilisation of healthy lifestyle in patents who attend the programme. METHODS: Patients in nine cardiology centres in Poland, who were treated for ACS between October and December 2014, were included. A dedicated questionnaire on healthy lifestyle and knowledge on cardiovascular risk factors was filled in by patients at hospital discharge. In January 2015 the same questionnaire was filled in by the same group of patients before their local Patient Club meeting. RESULTS: There were 1273 patients hospitalised for ACS who were invited to join the Patient Club initiative. Of them, 372 (29%) filled in the questionnaire both at discharge and at the Patient Club meeting. The percentage of patients who smoked cigarettes decreased from 14% to 5% (p < 0.001), and the number of those who had at least 30 min of physical activity daily increased from 50% to 58% (p = 0.015). CONCLUSIONS: Patients who attended Patient Club meetings usually four weeks after ACS showed significant benefit in terms of healthy lifestyle changes and more guideline-recommended management of cardiovascular risk factors.

Palladium(0)-catalysed synthesis of 2,3- and 3,4-unsaturated aryl ß-O-glycosides.

Arylation of 6-O-tert-butyldiphenylsilyl-3,4-di-O-isobutyloxycarbonyl-d-glucal (3) with various phenols in the presence of a catalytic amount of palladium(0) gave the corresponding 2,3- and 3,4-unsaturated ß-O-glycosides. The reaction is stereospecific, in all cases, only the ß-anomer is formed.

Haptically Guided Grasping. fMRI Shows Right-Hemisphere Parietal Stimulus Encoding, and Bilateral Dorso-Ventral Parietal Gradients of Object- and Action-Related Processing during Grasp Execution.

The neural bases of haptically-guided grasp planning and execution are largely unknown, especially for stimuli having no visual representations. Therefore, we used functional magnetic resonance imaging (fMRI) to monitor brain activity during haptic exploration of novel 3D complex objects, subsequent grasp planning, and the execution of the pre-planned grasps. Haptic object exploration, involving extraction of shape, orientation, and length of the to-be-grasped targets, was associated with the fronto-parietal, temporo-occipital, and insular cortex activity. Yet, only the anterior divisions of the posterior parietal cortex (PPC) of the right hemisphere were significantly more engaged in exploration of complex objects (vs. simple control disks). None of these regions were re-recruited during the planning phase. Even more surprisingly, the left-hemisphere intraparietal, temporal, and occipital areas that were significantly invoked for grasp planning did not show sensitivity to object features. Finally, grasp execution, involving the re-recruitment of the critical right-hemisphere PPC clusters, was also significantly associated with two kinds of bilateral parieto-frontal processes. The first represents transformations of grasp-relevant target features and is linked to the dorso-dorsal (lateral and medial) parieto-frontal networks. The second monitors grasp kinematics and belongs to the ventro-dorsal networks. Indeed, signal modulations associated with these distinct functions follow dorso-ventral gradients, with left aIPS showing significant sensitivity to both target features and the characteristics of the required grasp. Thus, our results from the haptic domain are consistent with the notion that the parietal processing for action guidance reflects primarily transformations from object-related to effector-related coding, and these mechanisms are rather independent of sensory input modality.

Partial correction of the CNS lysosomal storage defect in a mouse model of juvenile neuronal ceroid lipofuscinosis by neonatal CNS administration of an adeno-associated virus serotype rh.10 vector expressing the human CLN3 gene.

Juvenile neuronal ceroid lipofuscinosis (JNCL or CLN3 disease) is an autosomal recessive lysosomal storage disease resulting from mutations in the CLN3 gene that encodes a lysosomal membrane protein. The disease primarily affects the brain with widespread intralysosomal accumulation of autofluorescent material and fibrillary gliosis, as well as the loss of specific neuronal populations. As an experimental treatment for the CNS manifestations of JNCL, we have developed a serotype rh.10 adeno-associated virus vector expressing the human CLN3 cDNA (AAVrh.10hCLN3). We hypothesized that administration of AAVrh.10hCLN3 to the Cln3(Δex7/8) knock-in mouse model of JNCL would reverse the lysosomal storage defect, as well as have a therapeutic effect on gliosis and neuron loss. Newborn Cln3(Δex7/8) mice were administered 3 × 10(10) genome copies of AAVrh.10hCLN3 to the brain, with control groups including untreated Cln3(Δex7/8) mice and wild-type littermate mice. After 18 months, CLN3 transgene expression was detected in various locations throughout the brain, particularly in the hippocampus and deep anterior cortical regions. Changes in the CNS neuronal lysosomal accumulation of storage material were assessed by immunodetection of subunit C of ATP synthase, luxol fast blue staining, and periodic acid-Schiff staining. For all parameters, Cln3(Δex7/8) mice exhibited abnormal lysosomal accumulation, but AAVrh.10hCLN3 administration resulted in significant reductions in storage material burden. There was also a significant decrease in gliosis in AAVrh.10hCLN3-treated Cln3(Δex7/8) mice, and a trend toward improved neuron counts, compared with their untreated counterparts. These data demonstrate that AAVrh.10 delivery of a wild-type cDNA to the CNS is not harmful and instead provides a partial correction of the neurological lysosomal storage defect of a disease caused by a lysosomal membrane protein, indicating that this may be an effective therapeutic strategy for JNCL and other diseases in this category.

The antifungal properties of chicken egg cystatin against Candida yeast isolates showing different levels of azole resistance.

The increasing incidence of fungal infections together with the emergence of strains resistant to currently available antifungal drugs calls for the development of new classes of antimycotics. Naturally occurring antifungal proteins and peptides are of interest because of low toxicity, immunomodulatory potential and mechanisms of action distinct from those of currently available drugs. In this study, the potent antifungal activity of cystatin, affinity-purified from chicken egg white (CEWC), against the most frequent human fungal pathogens of the genus Candida was identified and characterised. CEWC inhibited the growth of azole-sensitive Candida albicans isolates with minimal inhibitory concentration (MIC) values ranging from 0.8 to 3.3 micromol l(-1), a potency comparable with those of fluconazole and histatin 5, the antimicrobial peptide of the human saliva. Similarly to histatin 5, CEWC activity was not compromised in azole-resistant isolates overproducing the multidrug efflux transporters Cdr1p and Cdr2p and did not antagonise fluconazole or amphotericin B. CEWC had candidacidal activity, as revealed by the time-kill assay, and, similarly to histatin 5, completely inhibited the growth at supra-MIC concentrations. This was in contrast to the fungistatic effect and trailing growth observed with fluconazole. CEWC inhibited the growth of Candida parapsilosis and Candida tropicalis at similar concentrations, whereas Candida glabrata was more resistant to CEWC.

A Kazal-type serine proteinase inhibitor from chicken liver (clTI-1): purification, primary structure, and inhibitory properties.

Low-molecular-mass trypsin inhibitor (clTI-1; chicken liver Trypsin Inhibitor-1) was purified from chicken liver by extraction with perchloric acid, ammonium sulfate precipitation, a combination of ethanol-acetone fractionation followed by gel filtration, ion-exchange chromatography and RP-HPLC on a C18 column. The inhibitor occurs in two isoforms with molecular masses of 5938.56 and 6026.29 Da (determined by MALDI TOFF mass spectrometry). The complete amino acid sequences of both isoforms were determined (UniProtKB/Swiss-Prot P85000; ISK1L_CHICK). The inhibitor shows a high homology to Kazal-type family inhibitors, especially to trypsin/acrosin inhibitors and pancreatic secretory trypsin inhibitors. clTI-1 inhibits both bovine and porcine trypsin (K(a)=1.1 x 10(9) M(-1) and 2.5 x 10(9) M(-1), respectively). Significant differences were shown in the inhibition of the anionic and cationic forms of chicken trypsin (K(a)=4.5 x 10(8) M(-1) and 1.2 x 10(10) M(-1)). Weak interaction with human plasmin (K(a)=1.2 x 10(7) M(-1)) was also revealed.