Ekstrakt metlike (Tamarix articulata) inhibira umnožavanje stanica hepatocelularnog karcinoma, potiče mehanizme stanične smrti i zaustavlja stanični ciklus u fazi G0/G1

Research background. From ancient times plants have been used for medicinal purposes against various ailments. In the modern era, plants are a major source of drugs and are an appealing drug candidate for the anticancer therapeutics against various molecular targets. Here we tested methanolic extract of dry leaves of Tamarix articulata for anticancer activity against a panel of hepatocellular carcinoma cells. Experimental approach. Cell viability of hepatocellular carcinoma cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay after a dose-dependent treatment with the extract of T. articulata. Phase-contrast microscopy and 4՛,6-diamidino-2-phenylindole (DAPI) staining served to analyse cellular and nuclear morphology. Immunoblotting was performed to determine the expression of proteins associated with autophagy, apoptosis and cell cycle. However, flow cytometry was used for the quantification of apoptotic cells and the analysis of cells in different phases of the cycle after the treatment with various doses of T. articulata. Additionally, acridine orange staining and 2՛,7՛-dichlorofluorescein diacetate (DCFH-DA) dye were used to analyse the quantification of autophagosomes and reactive oxygen species. Results and conclusion. Our results demonstrate that T. articulata methanolic extract exhibits promising antiproliferative activity with IC50 values (271.1±4.4), (298.3±7.1) and (336.7±6.1) µg/mL against hepatocellular carcinoma HepG2, Huh7D12 and Hep3B cell lines, respectively. Mechanistically, we found that T. articulata methanolic extract induces cell death by activating apoptosis and autophagy pathways. First, T. articulata methanolic extract promoted autophagy, which was confirmed by acridine orange staining. The immunoblotting analysis further confirmed that the extract at higher doses consistently induced the conversion of LC3I to LC3II form with a gradual decrease in the expression of autophagy substrate protein p62. Second, T. articulata methanolic extract promoted reactive oxygen species production in hepatocellular carcinoma cells and activated reactive oxygen species-mediated apoptosis. Flow cytometry and immunoblotting analysis showed that the plant methanolic extract induced dose-dependent apoptosis and activated proapoptotic proteins caspase-3 and PARP1. Additionally, the extract triggered the arrest of the G0/G1 phase of the cell cycle and upregulated the protein expression of p27/Kip and p21/Cip, with a decrease in cyclin D1 expression in hepatocellular carcinoma cells. Novelty and scientific contribution. The current study demonstrates that T. articulata methanolic extract exhibits promising anticancer potential to kill tumour cells by programmed cell death type I and II mechanisms and could be explored for potential drug candidate molecules to curtail cancer in the future.Pozadina istraživanja. Biljke se od davnina koriste u medicinske svrhe za liječenje različitih bolesti. U moderno doba glavni su izvor sirovina za proizvodnju lijekova, pa tako i onih protiv različitih oblika karcinoma. U ovom smo radu ispitali antikancerogeno djelovanje metanolnog ekstrakta lišća metlike (Tamarix articulata) na niz stanica hepatocelularnog karcinoma. Eksperimentalni pristup. Preživljavanje stanica hepatocelularnog karcinoma ovisno o dozi ekstrakta metlike utvrđeno je pomoću tzv. MTT testa odnosno bojanjem (3-(4,5-dimetiltiazol-2-ilo)-2,5-difeniltetrazol bromidom. Stanična i nuklearna morfologija ispitane su pomoću mikroskopa s faznim kontrastom i bojanjem 4\u27,6-diamidino-2-fenilindol dihidrokloridom (DAPI). Imunoblot analizom određena je ekspresija proteina povezanih s autofagijom, apoptozom i staničnim ciklusom. S druge strane, protočnom je citometrijom određen broj apoptotičnih stanica nakon obrade različitim koncentracijama ekstrakta metlike, te su izbrojane stanice u različitim fazama staničnog ciklusa. Uz to, bojanjem akridinskom narančastom i diacetatnim oblikom 2\u27,7\u27-diklorofluoresceina (DCFHDA) određena je količina autofagosoma i reaktivnih kisikovih spojeva. Rezultati i zaključci. Dobiveni rezultati pokazuju da metanolni ekstrakt metlike ima dobar antiproliferacijski učinak, s IC50 vrijednostima od (271,1±4,4) µg/mL za staničnu liniju HepG2, (298,3±7,1) µg/mL za staničnu liniju Huh7D12 i (336,7±6,1) µg/mL za staničnu liniju Hep3B. Utvrdili smo da ekstrakt inducira staničnu smrt aktivacijom apoptoze i autofagije. Prvo je aktivirao autofagiju, što je potvrđeno bojanjem akridinskom narančastom. Imunoblot analizom je dodatno potvrđeno da veće koncentracije ekstrakta induciraju konverziju LC3I u LC3II, te postepeno smanjuju ekspresiju proteina p62. Zatim je ekstrakt potaknuo nastajanje reaktivnih kisikovih spojeva u stanicama hepatocelularnog karcinoma te time aktivirao apoptozu. Protočna citometrija i imunoblot analiza pokazale su da je ekstrakt metlike ovisno o dozi inducirao apoptozu i aktivirao proapoptotičke proteine kaspazu-3 i PARP1. Osim toga, zaustavio je stanični ciklus u fazi G0/G1 te ushodno regulirao ekspresiju proteina p27/Kip i p21/Cip, pritom smanjujući ekspresiju ciklina D1 u stanicama hepatocelularnog karcinoma. Novina i znanstveni doprinos. U radu smo pokazali da metanolni ekstrakt metlike ima dobar potencijal za ubijanje stanica tumora mehanizmima programirane smrti stanica tipa I i II, te da se može upotrijebiti za pronalazak molekula koje se mogu upotrijebiti u proizvodnji lijekova protiv tumora

DNA Transfer in Forensic Science: Recent Progress towards Meeting Challenges.

Understanding the factors that may impact the transfer, persistence, prevalence and recovery of DNA (DNA-TPPR), and the availability of data to assign probabilities to DNA quantities and profile types being obtained given particular scenarios and circumstances, is paramount when performing, and giving guidance on, evaluations of DNA findings given activity level propositions (activity level evaluations). In late 2018 and early 2019, three major reviews were published on aspects of DNA-TPPR, with each advocating the need for further research and other actions to support the conduct of DNA-related activity level evaluations. Here, we look at how challenges are being met, primarily by providing a synopsis of DNA-TPPR-related articles published since the conduct of these reviews and briefly exploring some of the actions taken by industry stakeholders towards addressing identified gaps. Much has been carried out in recent years, and efforts continue, to meet the challenges to continually improve the capacity of forensic experts to provide the guidance sought by the judiciary with respect to the transfer of DNA

DNA transfer in forensic science: A review

© 2018 Elsevier B.V. Understanding the variables impacting DNA transfer, persistence, prevalence and recovery (DNA-TPPR) has become increasingly relevant in investigations of criminal activities to provide opinion on how the DNA of a person of interest became present within the sample collected. This review considers our current knowledge regarding DNA-TPPR to assist casework investigations of criminal activities. There is a growing amount of information available on DNA-TPPR to inform the relative probabilities of the evidence given alternative scenarios relating to the presence or absence of DNA from a specific person in a collected sample of interest. This information should be used where relevant. However, far more research is still required to better understand the variables impacting DNA-TPPR and to generate more accurate probability estimates of generating particular types of profiles in more casework relevant situations. This review explores means of achieving this. It also notes the need for all those interacting with an item of interest to have an awareness of DNA transfer possibilities post criminal activity, to limit the risk of contamination or loss of DNA. Appropriately trained forensic practitioners are best placed to provide opinion and guidance on the interpretation of profiles at the activity level. However, those requested to provide expert opinion on DNA-related activity level issues are often insufficiently trained to do so. We advocate recognition of DNA activity associated expertise to be distinct from expertise associated with the identification of individuals. This is to be supported by dedicated training, competency testing, authorisation, and regular fit for purpose proficiency testing. The possibilities for experts to report on activity-related issues will increase as our knowledge increases through further research, access to relevant data is enhanced, and tools to assist interpretations are better exploited. Improvement opportunities will be achieved sooner, if more laboratories and agencies accept the need to invest in these aspects as well as the training of practitioners

The Tabby cat locus maps to feline chromosome B1.

The Tabby markings of the domestic cat are unique coat patterns for which no causative candidate gene has been inferred from other mammals. In this study, a genome scan was performed on a large pedigree of cats that segregated for Tabby coat markings, specifically for the Abyssinian (Ta-) and blotched (tbtb) phenotypes. There was linkage between the Tabby locus and eight markers on cat chromosome B1. The most significant linkage was between marker FCA700 and Tabby (Z = 7.56, theta = 0.03). Two additional markers in the region supported linkage, although not with significant LOD scores. Pairwise analysis of the markers supported the published genetic map of the cat, although additional meioses are required to refine the region. The linked markers cover a 17-cM region and flank an evolutionary breakpoint, suggesting that the Tabby gene has a homologue on either human chromosome 4 or 8. Alternatively, Tabby could be a unique locus in cats

Prolapsus Gravidique: Facteurs de Risque, Complications et Prise en Charge en Afrique Sub-Saharienne

Objectif :  Le but de cette étude était d’aborder les caractéristiques socio démographie, les aspects thérapeutiques et le pronostic du prolapsus utérin extériorisé survenu pour la première fois pendant la grossesse. Patientes et Méthodes : Il s’agissait d’une étude prospective sur une période de 12 mois (1er janvier au 31 décembre 2021) portant sur les patientes prise en charge pour prolapsus gestationnel. Résultats: Sept cas ont été enregistrés durant la période d’étude. La fréquence du prolapsus gestationnel était de 2 cas/1000 accouchements dans notre service. Les patientes étaient jeunes (28-31 ans), multipares avec un âge moyen de 28,57 ans et une parité moyenne de quatre enfants. Elles étaient très jeunes à leurs premier accouchement (16-20 ans). La plupart de facteurs de risques décrits dans la littérature étaient retrouvés chez nos patientes. La rupture prématurée des membranes (2 cas, 28,57%), la chorioamniotite (1 cas, 14,28%), l’accouchement prématuré (1 cas, 14,28%), l’hémorragie de la délivrance (1 cas, 14,28%) et l’anémie (1 cas, 14,28%) ont été les complications retrouvées. Un traitement conservateur a été réalisé chez toutes les patientes (100%). L’évolution était favorable avec une régression spontanée du prolapsus dans le post-partum immédiat chez toutes nos patientes (100%). Après six mois de suivi, les prolapsus n´ont pas récidivés. Conclusion : Le prolapsus gestationnel n’est pas exceptionnel dans notre contexte. Malgré son caractère angoissant pour la patiente, sa famille et l’équipe obstétricale, une attitude conservatrice peut se discuter devant un prolapsus apparu pour la première fois au cours de la grossesse chez une patiente jeune sans antécédents pathologiques particuliers.   Objective : The aim of this study was to address the socio-demographic characteristics, therapeutic aspects and prognosis of externalized uterine prolapse occurring for the first time during pregnancy. Patients and Methods : This was a prospective study over a period of 12 months (January 1 to December 31, 2021) focusing on patients treated for gravidarum prolapse. Results : Seven cases were recorded during the study period. The frequency of gravidarum prolapse was 2 cases/1000 deliveries in our department. The patients were young (28-31 years old), multiparous with an average age of 28.57 years and an average parity of four children. They were very young when they first gave birth (16-20 years old). Most of the risk factors described in the literature were found in our patients. Premature rupture of membranes (2 cases, 28.57%), chorioamnionitis (1 case, 14.28%), premature delivery (1 case, 14.28%), postpartum hemorrhage (1 case, 14.28%) and anemia (1 case, 14.28%) were the complications found. Conservative treatment was carried out in all patients (100%). The evolution was favorable with spontaneous regression of the prolapse in the immediate postpartum period in all our patients (100%). After six months of follow-up, the prolapses have not recurred. Conclusion : gravidarum prolapse is not exceptional in our context. Despite its distressing nature for the patient, her family and the obstetric team, a conservative attitude can be questioned when faced with a prolapse appearing for the first time during pregnancy in a young patient without any particular pathological history

The role of 5-HT2C receptors in touchscreen visual reversal learning in the rat: a cross-site study.

RATIONALE: Reversal learning requires associative learning and executive functioning to suppress non-adaptive responding. Reversal-learning deficits are observed in e.g. schizophrenia and obsessive-compulsive disorder and implicate neural circuitry including the orbitofrontal cortex (OFC). Serotonergic function has been strongly linked to visual reversal learning in humans and experimental animals but less is known about which receptor subtypes are involved. OBJECTIVES: The objectives of the study were to test the effects of systemic and intra-OFC 5-HT2C-receptor antagonism on visual reversal learning in rats and assess the psychological mechanisms underlying these effects within novel touchscreen paradigms. METHODS: In experiments 1-2, we used a novel 3-stimulus task to investigate the effects of 5-HT2C-receptor antagonism through SB 242084 (0.1, 0.5 and 1.0 mg/kg i.p.) cross-site. Experiment 3 assessed the effects of SB 242084 in 2-choice reversal learning. In experiment 4, we validated a novel touchscreen serial visual reversal task suitable for neuropharmacological microinfusions by baclofen-/muscimol-induced OFC inactivation. In experiment 5, we tested the effect of intra-OFC SB 242084 (1.0 or 3.0 μg/side) on performance in this task. RESULTS: In experiments 1-3, SB 242084 reduced early errors but increased late errors to criterion. In experiment 5, intra-OFC SB 242084 reduced early errors without increasing late errors in a reversal paradigm validated as OFC dependent (experiment 4). CONCLUSION: Intra-OFC 5-HT2C-receptor antagonism decreases perseveration in novel touchscreen reversal-learning paradigms for the rat. Systemic 5-HT2C-receptor antagonism additionally impairs late learning-a robust effect observed cross-site and potentially linked to impulsivity. These conclusions are discussed in terms of neural mechanisms underlying reversal learning and their relevance to psychiatric disorders.The research leading to these results has received support from the Innovative Medicine Initiative Joint Undertaking under grant agreement no. 115008 of which resources are composed of EFPIA in-kind contribution and financial contribution from the European Union’s Seventh Framework Programme (FP7/2007-2013). The study was also supported by a Wellcome Trust Grant (089589/z/09/z) to T.W.R., B.J. Everitt, B.J. Sahakian, A.C. Roberts and J.W. Dalley, and by a Wellcome Trust Senior Investigator Award to T.W.R. (104631/Z/14/Z). The Behavioural and Clinical Neuroscience Institute is co-funded by the Medical Research Council and the Wellcome Trust. J.A. was supported by the Swedish Pharmaceutical Society and the Swedish Research Council (350-2012-230). S.R.O.N was supported by BBSRC and Eli Lilly through CASE studentship (BB/F529054/1).This is the final version of the article. It first appeared from Springer via http://dx.doi.org/10.1007/s00213-015-3963-

Transport properties of dense fluid argon

We calculate using molecular dynamics simulations the transport properties of realistically modeled fluid argon at pressures up to $\simeq 50GPa$ and temperatures up to $3000K$. In this context we provide a critique of some newer theoretical predictions for the diffusion coefficients of liquids and a discussion of the Enskog theory relevance under two different adaptations: modified Enskog theory (MET) and effective diameter Enskog theory. We also analyze a number of experimental data for the thermal conductivity of monoatomic and small diatomic dense fluids.Comment: 8 pages, 6 figure

The effects of soaking for DNA recovery on the striation patterns of fired cartridge cases

© 2019, © 2019 Australian Academy of Forensic Sciences. The recovery of trace DNA from fired cartridge cases has recently gained increased interest throughout the literature, with a variety of methods currently being explored. Soaking fired cartridge cases in a lysis buffer holds potential in producing meaningful DNA profiles; however, chemical interactions between the lysis buffer and brass cartridge cases may limit the efficacy of this method. This preliminary study examines the effects of soaking on the microscopic striation detail of brass and nickel 9 mm Parabellum (9 mmP) calibre and.22 Long Rifle (.22LR) calibre fired cartridge cases. Headstamp and coarse striation patterns on 9 mmP fired cartridge cases and finer striation patterns along the outer wall of.22LR fired cartridge cases were microscopically examined prior to and following soaking. Soaking was performed by submerging the fired cartridge cases in 380 µl of ATL buffer (Qiagen, Germany) for 20 minutes. Microscopic analysis of brass and nickel 9 mmP and.22LR fired cartridge cases showed that coarse and fine striation detail remain unaffected following soaking. These results indicate that comparative ballistics examinations may be performed following DNA recovery using the soaking method

Diffusion Time-Scale Invariance, Markovization Processes and Memory Effects in Lennard-Jones Liquids

We report the results of calculation of diffusion coefficients for Lennard-Jones liquids, based on the idea of time-scale invariance of relaxation processes in liquids. The results were compared with the molecular dynamics data for Lennard-Jones system and a good agreement of our theory with these data over a wide range of densities and temperatures was obtained. By calculations of the non-Markovity parameter we have estimated numerically statistical memory effects of diffusion in detail.Comment: 10 pages, 3 figure

Evaluation of Antiproliferative Properties of CoMnZn-Fe₂O₄ Ferrite Nanoparticles in Colorectal Cancer Cells

The PEG-coated ferrite nanoparticles Co0.2Mn0.6Zn0.2Fe2O4 (X1), Co0.4Mn0.4Zn0.2Fe2O4 (X2), and Co0.6Mn0.2Zn0.2Fe2O4 (X3) were synthesized by the coprecipitation method. The nanoparticles were characterized by XRD, Raman, VSM, XPS, and TEM. The magnetic hyperthermia efficiency (MH) was determined for PEG-coated nanoparticles using an alternating magnetic field (AMF). X2 nanoparticles displayed the highest saturation magnetization and specific absorption rate (SAR) value of 245.2 W/g for 2 mg/mL in a water medium. Based on these properties, X2 nanoparticles were further evaluated for antiproliferative activity against HCT116 cells at an AMF of 495.25 kHz frequency and 350 G strength, using MTT, colony formation, wound healing assays, and flow cytometry analysis for determining the cell viability, clonogenic property, cell migration ability, and cell death of HCT116 cells upon AMF treatment in HCT116 cells, respectively. We observed a significant inhibition of cell viability (2% for untreated control vs. 50% for AMF), colony-forming ability (530 cells/colony for untreated control vs. 220 cells/colony for AMF), abrogation of cell migration (100% wound closure for untreated control vs. 5% wound closure for AMF), and induction of apoptosis-mediated cell death (7.5% for untreated control vs. 24.7% for AMF) of HCT116 cells with respect to untreated control cells after AMF treatment. Collectively, these results demonstrated that the PEG-coated (CoMnZn-Fe2O4) mixed ferrite nanoparticles upon treatment with AMF induced a significant antiproliferative effect on HCT116 cells compared with the untreated cells, indicating the promising antiproliferative potential of the Co0.4Mn0.4Zn0.2Fe2O4 nanoparticles for targeting colorectal cancer cells. Additionally, these results provide appealing evidence that ferrite-based nanoparticles using MH could act as potential anticancer agents and need further evaluation in preclinical models in future studies against colorectal and other cancers.</p