A Role for the Circadian Clock in Colorectal Cancer Progression: A Comprehensive Molecular Analysis on the Interplay between Core-Clock Genes and Metastasis-related Cellular Processes

Störungen der zirkadianen Uhr beeinflussen zelluläre Prozesse wie Transkription, Zellzyklus und Stoffwechsel und können Tumorentstehung fördern. Unsere Studien untersuchten die Auswirkungen solcher Störungen auf die Krebsentwicklung durch Herunterregulation oder Ausschalten von Kern-Uhrgenen wie BMAL1 (ARNTL), PER2 oder NR1D1. Wir verwendeten in vitro- und in vivo-Modellen mit Darmkrebszelllinien HCT116, SW480 und SW620. Diese Manipulationen veränderten die zirkadiane Expression von MYC, WEE1 und TP53 (wichtig für Zellzyklus und Apoptose) sowie MACC1 (verbunden mit epithelial-mesenchymaler Transition und Metastasierung). Diese Veränderungen beeinflussen Zellproliferation, Apoptose, Migration und Invasion. Das Ausschalten von NR1D1 verringerte die Zellbeweglichkeit in vitro und reduzierte die Mikrometastasenbildung in vivo, begleitet von geänderten SNAI1 und CD44-Expressionen. MACC1 wird in HCT116-Wildtypzellen zirkadian exprimiert, was nach dem Ausschalten der Kern-Uhrgene gestört war. Wir identifizierten außerdem eine MACC1-NR1D1-Protein-Interaktion, die eine neue Regulierungsachse bei der Darmkrebsprogression darstellt. Unsere Daten zeigen, dass MACC1-Modulation den zirkadianen Phänotyp und Krebsfortschritt beeinflusst. MACC1-Knockout reduzierte die BMAL1-Oszillationsperiode, während seine Überexpression den gegenteiligen Effekt hatte. Dieses Zusammenspiel unterstreicht die Komplexität der Mechanismen bei Darmkrebs und die Rolle der zirkadianen Uhr bei der Metastasierung. Unsere Ergebnisse heben die Rolle von Kern-Uhrgenen bei Krebsprozessen wie Migration und Invasion hervor und bieten Einblicke in das MACC1-zirkadiane Uhr-Zusammenspiel in Darmkrebs. Zukünftige Forschungen könnten chronotherapeutische Strategien entwickeln, um Krebsbehandlungen zu personalisieren und zu verbessern.Disruptions of the circadian clock affect cellular processes like transcription, cell cycle, and metabolism, potentially triggering tumorigenesis. Our studies examined the effects of these disruptions on cancer by downregulating or knocking out core-clock genes BMAL1 (ARNTL), PER2, or NR1D1. For this, we used in vitro and in vivo models with colorectal cancer (CRC) cell lines HCT116, SW480, and SW620. Core-clock gene manipulations altered circadian expression of MYC, WEE1, TP53 (involved in cell cycle and apoptosis), and MACC1 (linked to epithelial-mesenchymal transition and metastasis formation), affecting proliferation, apoptosis, migration, and invasion. NR1D1 knockdown reduced cell motility in vitro and decreased micrometastasis formation in vivo, with altered SNAI1 and CD44 expression. Furthermore, we showed that MACC1 is circadian expressed in HCT116 wild-type cells, and that its rhythmic expression is disrupted after core-clock gene knockout. We also found MACC1-NR1D1 protein-protien interactions, suggesting a new regulatory axis in CRC progression. Our data show MACC1 modulation impacts the circadian clock phenotype and cancer progression. Remarkably, MACC1 knockout reduced BMAL1 promoter oscillation period, while its overexpression had the opposite effect. This interplay highlights the circadian clock complexity and its role in CRC metastasis. Our findings underscore vital roles for core-clock genes in cancer processes like migration and invasion, providing insights into MACC1-circadian clock interplay in CRC. Future research may develop chronotherapeutic strategies for more personalized and effective treatments

Studies of Adenosine and GABAA Receptor Functions in Rat Hippocampal Slices

Recent evidence has indicated that adenosine, in addition to potassium and calcium currents, may also affect chloride movement in hippocampal neurones. This project was undertaken to determine the possible role of adenosine on chloride channels and synaptic plasticity in comparison with a selective GABA[A] agonist, muscimol. Extracellular recordings were made from the CA1 pyramidal cell layer of hippocampal slices in response to stimulation of Schaffer collateral fibres in stratum radiatum (0.01 Hz). Adenosine and muscimol induced concentration dependent reductions in the amplitude of orthodromically induced population potentials. In order to eliminate effects of these agents on potassium channels, experiments were performed in the presence of barium, 1mM (in some experiments in the presence of 1 mM tolbutamide). This concentration increased potential size, and reduced the inhibitory effect of adenosine on population spike size, but not synaptic potential size. This profile is consistent with the blockade of potassium channels associated only with the postsynaptic effects of adenosine. However, muscimol responses were unaffected. Adenosine potentiated the ability of muscimol to inhibit evoked potentials in the absence or presence of barium. The potentiation was prevented by the Al selective antagonist 8-cyclopentyltheophylline. The effects of adenosine, as well as muscimol, were reduced by the chloride channel blocker DIDS, which also prevented the adenosine potentiation of muscimol. The results indicate the ability of adenosine to operate chloride channels in hippocampal neurones, and suggest a potentiative interaction between adenosine and muscimol which also involve chloride channels. The second part of this study was to examine neurosteroids which have been reported to be positive modulators of the GABAA receptors. Alphaxalone and 5alpha-pregnan-3alpha-o1-20-one potentiated the inhibitory effect of muscimol on the population spike size at low concentrations (0.5 and 1muM) that had no significant effect on the spike size by themselves. This profile is in agreement with other reports which have described the effect of these neurosteroids as barbiturate-like. Alphaxalone and 5alpha-pregnan-3alpha-o1-20-one also at low concentrations potentiated the inhibitory effect of adenosine alone and in the presence of barium 1 mM which blocked adenosine activated potassium channels. Alphaxalone failed to potentiate the inhibitory effect of adenosine in the presence of bicuculline 1muM. It is concluded that these neurosteroids enhanced the potentiative interaction between adenosine and muscimol in the presence of barium. The results indicate that adenosine's effects are normally enhanced by virtue of the potentiative interaction occurring with endogenous GABA. In addition to this, the results show the chloride channels activated by adenosine to be different from those operated by the GABA[A] receptor. The third part of this project was to investigate the role of GABA[A] and adenosine receptors on long-term depression (LTD) and synaptic plasticity. Unlike long-term potentiation, LTD in the central nervous system remains poorly understood. Muscimol induced a time and concentration-dependent LTD in the amplitude of orthodromic potentials. Increasing the stimulation frequency from 0.01 Hz to 1 Hz for 10 seconds reversed the LTD induced by muscimol. Although adenosine decreased the spike size in a concentration-dependent manner, it failed to induce LTD. Muscimol also induced LTD in the absence of electrical stimulation. Alphaxalone and 5alpha-pregnan-3alpha-o1-20-one at concentrations that did not have any effect themselves on the population spike (0.5 and 1 muM), potentiated the inhibitory effect of muscimol on the population spike size. These neurosteroids at high concentrations (5 and 10 muM) decreased the spike size by themselves. On the other hand, at the low concentrations both steroids were able to potentiate the ability of muscimol to induce LTD. Moreover, muscimol 1 muM which is not able to induce LTD, alphaxalone and 5alpha-pregnan-3alpha-o1-20-one 1 muM maintained the LTD induced by muscimol 10 muM. Bicuculline 5 muM reversed the LTD induced by muscimol 10 muM. To examine the possible role of glutamate receptors in LTD induced by muscimol a number of NMDA and metabotropic receptors agonists and antagonists were used. The NMDA receptor antagonist 2-AP5, the NMDA/metabotropic antagonist 2-AP3 and selective metabotropic antagonist L(+)-AP3 failed to modify the LTD. Quisqualic acid and (1S,3R)- aminocyclopentane dicarboxylic acid (ACPD), a selective agonist at metabotropic receptors, did not induce LTD or short-term depression, whereas kynurenic acid prevented the reversal of the LTD obtained at 1 Hz. It is concluded that LTD can be induced by the selective activation of GABA[A] receptors and the lack of involvement of glutamate receptors in the protocol which is presented in this study confirms the unique role of classical GABA[A] receptors in the effect of muscimol and may indicate a novel type of long-lasting depression. Furthermore, the failure of adenosine to induce LTD taken together with the earlier results, suggests that the adenosine activated chloride channel differs from the GABA[A] receptor chloride channel

Preparation of Liposomal Nanoparticles Containing Ziziphora tenuir Essential Oil and Evaluation of its Antimicrobial Effects

Background and Objectives  Natural preservatives extracted from herbs are important sources for bioactive compounds that can be used in protection of food products. Essential oils are aromatic oily liquids, obtained from plant material like flowers, buds, seeds, leaves, and roots. Unfortunately, most natural compounds are biologically instable, poorly soluble in water and they distribute poorly to target sites. Currently, some novel methods have been introduced in order to improve their stability and their bioavailability, among which is the use of liposomal encapsulation. Microencapsulation reduces reactivity with the environment (water, oxygen, light), decreases the evaporation or the transfer rate to the outside environment, promotes handling ability, masks taste and enhances dilution to achieve a uniform distribution in the final product when used in very small amounts. Essential oils, as natural extracted compounds extracted from plants, are unstable compounds with low water solubility and unable to achieve target cells. Essential oils encapsulation by nanoliposomes is a novel method for increasing their biological activity and protecting them from destructive factors. The aim of this study was production and optimization of nanoliposomes containing Z. teniur essential oil and investigating their antibacterial effects against pathogens (Staphylococcus aureus and Escherichia coli).   Materials and Methods  Lipid film hydration method was used to produce nanoliposomes containing Z. teniur essential oil. Soy phosphatidylcholine and cholesterol were the main wall materials and chloroform was used as the mixing solvent . The particle size of nanoliposomes and their zeta-potential were investigated using laser diffraction method. In order to determine the minimum inhibitory concentration and the minimum bactericidal concentration of Z. teniur essential oil against examined bacteria, serial dilution method was used. Also, antioxidant activity of free and nano-encapsulated essential oil of Z. teniur was determined by DPPH method.   Results  According to the results, highest encapsulation efficiency achieved by using 80:20 ratio of soy phosphatidylcholine to cholesterol in nanoliposomes’ wall structures. In general, by increasing the ratio of phosphatidylcholine to cholesterol, encapsulation efficiency was improved. Zeta-potential of nanoliposomes was equal to -5.3 mv and mean particle sizes were in the range of 94.7-119.9 nm. Results indicated that essential oil ejection from nanoliposomes has direct relation to the time of storage and after 30 hours, ejection rate will increase considerably. Ejection rate was higher in phosphate buffer pH=7.4 in comparison with phosphate buffer pH=5.4. Minimum inhibitory concentration and minimum bactericidal concentration of free essential oil against Escherichia coli was 100 and 175 (µl/ml) respectively. Although, Minimum inhibitory concentration and minimum bactericidal concentration of nanoliposomes containing Z. teniur essential oil were equal to 75 and 150 (µl/ml) respectively. Also, results shown that , minimum inhibitory concentration and minimum bactericidal concentration of encapsulated Z. teniur essential oil against Staphylococcus aureus were lower in comparison with free form of Z. teniur essential oil. Staphylococcus aureus (as Gram-positive bacteria) was more susceptible than Escherichia coli (as Gram-negative bacteria).   Conclusion  Encapsulation of Z. teniur essential oil by nanoliposomes led to improve antibacterial effects of essential oil against Staphylococcus aureus and Escherichia coli. Also, investigating of antioxidant activity showed that encapsulated Z. teniur essential oil in nanoliposomes was more effective than free form of Z. teniur essential oil in scavenging of DPPH free radicals. Using nanoliposome encapsulation technology can be an effective way for increasing the efficiency of natural antibacterial compounds and essential oils encapsulated in nanoliposomes are suitable alternatives for synthetic preservatives used in food industry nowadays. The use of liposomes containing Z. teniur essential oil can provide the necessary protection against growth of spoilage and pathogenic microorganisms such as Staphylococcus aureus and Escherichia coli in food products

Evaluation of the chemical composition and in vitro antimicrobial activity of Rosmarinus officinalis, Zataria multiflora, Anethum graveolens and Eucalyptus globulus against Streptococcus iniae; the cause of zoonotic disease in farmed fish

There is a growing interest of industry to replace synthetic chemicals by natural products with bioactive properties from plant origin. The purpose of this study was to elucidate the factors affecting antimicrobial effectiveness of essential oils Rosmarinus officinalis, Zataria multiflora, Anethum graveolens and Eucalyptus globulus against food spoilage and pathogenic bacteria, Streptococcus iniae; the cause of zoonotic streptococcosis in fish. Food conservation is based on an intermittent search for foods with a high nutritional quality and microbial stability and it has been reached by the control of the growth/survival of spoiling and pathogen foodborne microorganisms. Based on several reports, fish streptococcosis is currently considered as one of the main limiting factors in the aquaculture industry, due to the significant economic losses (annually more than $150 million) that these infections cause in different cultured fresh and seawater fish species worldwide. The sensitivity of S. iniae to antibacterial activity of the essential oils was determined using well diffusion assays and paper disc diffusion method. The ranges of minimum inhibitory concentration (MIC) of the oils and extracts were 3.9-250 and 7.8-500 µg/ml and the ranges of minimum bactericidal concentration (MBC) values for the oils and extracts were found to be in the range of 7.8-250 and 15.6-500 µg/ml, respectively. The essential oils exhibited antibacterial activity against S. iniae. The essential oil of rosemary showed the strongest antimicrobial activity

The Effect of Aqueous Extract of Saffron (Crocus sativus L. Stigma ) on the Behavior of Salmonella Typhimurium in A Food Model during Storage at Different Temperatures

Background: Given the concerns about the use of chemical preservatives in food, the consumers and producers have been interested in natural alternatives, such as plant essential oils and extracts. Since there are limited studies about the effect of saffron (Crocus sativus L.) on the behavior of foodborne pathogens in food models, this study aimed to determine the inhibitory effect of aqueous extract of saffron stigma on the growth behavior of Salmonella Typhimurium (S. Typhimurium) in commercial barley soup (as a food model) during storage at different temperatures. Method s : The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the extract were determined against S. Typhimurium using broth microdilution method. The growth of S . Typhimurium was investigated in the presence of this extract in commercial barley soup during 12 days of storage at 10, 20, and 30 °C. Results: The MIC and MBC values for saffron extract against S . Typhimurium were 100 and >200 mg/m l , respectively. Also, the saffron extract at a concentration of 200 mg/ml and temperature of 10 °C had the highest inhibitory effect on the growth of bacteria in commercial barley soup during storage. Conclusion: According to the results of this study, the antimicrobial effect of this extract increased in a dose -dependent manner against this bacterium. Therefore, the use of proper concentrations of this extract together with appropriate storage temperature can have an appropriate inhibitory effect on the growth of this bacterium, improving food safety shelf life

Self-help interventions for depressive disorders and depressive symptoms: a systematic review

<p>Abstract</p> <p>Background</p> <p>Research suggests that depressive disorders exist on a continuum, with subthreshold symptoms causing considerable population burden and increasing individual risk of developing major depressive disorder. An alternative strategy to professional treatment of subthreshold depression is population promotion of effective self-help interventions that can be easily applied by an individual without professional guidance. The evidence for self-help interventions for depressive symptoms is reviewed in the present work, with the aim of identifying promising interventions that could inform future health promotion campaigns or stimulate further research.</p> <p>Methods</p> <p>A literature search for randomised controlled trials investigating self-help interventions for depressive disorders or depressive symptoms was performed using PubMed, PsycINFO and the Cochrane Database of Systematic Reviews. Reference lists and citations of included studies were also checked. Studies were grouped into those involving participants with depressive disorders or a high level of depressive symptoms, or non-clinically depressed participants not selected for depression. A number of exclusion criteria were applied, including trials with small sample sizes and where the intervention was adjunctive to antidepressants or psychotherapy.</p> <p>Results</p> <p>The majority of interventions searched had no relevant evidence to review. Of the 38 interventions reviewed, the ones with the best evidence of efficacy in depressive disorders were S-adenosylmethionine, St John's wort, bibliotherapy, computerised interventions, distraction, relaxation training, exercise, pleasant activities, sleep deprivation, and light therapy. A number of other interventions showed promise but had received less research attention. Research in non-clinical samples indicated immediate beneficial effects on depressed mood for distraction, exercise, humour, music, negative air ionisation, and singing; while potential for helpful longer-term effects was found for autogenic training, light therapy, omega 3 fatty acids, pets, and prayer. Many of the trials were poor quality and may not generalise to self-help without professional guidance.</p> <p>Conclusion</p> <p>A number of self-help interventions have promising evidence for reducing subthreshold depressive symptoms. Other forms of evidence such as expert consensus may be more appropriate for interventions that are not feasible to evaluate in randomised controlled trials. There needs to be evaluation of whether promotion to the public of effective self-help strategies for subthreshold depressive symptoms could delay or prevent onset of depressive illness, reduce functional impairment, and prevent progression to other undesirable outcomes such as harmful use of substances.</p

Effect of Rosmarinus officinalis essential oil and nisin on Log P% of Streptococcus iniae in BHI broth

Background and Aim: Streptococcus is one of the most important food-borne diseases and zoonotic that caused by streptococcus species especially S.iniae. The present study was undertaken to evaluate the effects of R. officinalis essential oil, nisin, temperature, pH and storage time (43 days) on the log10 probability percentage of growth initiation (log P %) of S.iniae in brain heart infusion (BHI) broth in a factorial design study. Materials and Methods: The essential oil yield of the air-dried material was analyzed by gas chromatography (GC) (Agilent 6890, UK). Lyophilized cultures of S. iniae obtained from Department of Aquatic Animals Health and Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran, were used in this study. The log P% was calculated from the number of total tubes (out of 24) for each EO–N–pH–T combination showing visible growth up to a certain observation time, using 3×8 most probable number (MPN) method. Results: Concentrations of essential oil with 0.75 µg/ml nisin can inhibit growth of bacterial and log P% was calculated as -4.241in 37 ºC. The synergistic effect 0.25 µg/ml of nisin with concentrations of essential oil (0.005 and 0.015%) was observed in 15 ºC, thus no significantly affected (P>0.01) with 0.75 µg/ml of nisin. The growth of bacterial was completely inhibited at combinations of 0.0015% essential oil with 0.75 µg/ml of nisin, 4 ºC and pH= 5.5 during storage times. Conclusions: The synergistic effect of R. officinalis essential oil and nisin could increase the scope of essential oil usage within the food industry

Nanoencapsulation of green tea extract by thin film layer method and its properties

The application of natural compounds including green tea extract (GTE) in food preparation and pharmaceutical industries is limited. Encapsulation in nanoliposomes could be used as a delivery system to protect these compounds during processing and storage. In this study physicochemical characterization, total phenol content and antibacterial and antioxidant activity of green tea extract encapsulated in nanoliposomes were evaluated. GTE was encapsulated in liposomes by thin film layer method and reached to nanoscale with sonication. The antioxidant activity of nanoliposomal GTE was estimated by DPPH assay. The antibacterial activity of nanoliposomal GTE against Bacillus cereus (ATCC11778), Salmonella typhimurium 138 phage type 2, Escherichia coli O157:H7 and Listeria monocytogenes (ATCC19118) was determined using well diffusion technique. The mean diameter of nanoliposomes was about 44.7±1.9 nm and had 0.203±0.014 polydispersity index. Entrapment efficiency of nanoliposomal GTE under the optimum conditions was 97%. Antibacterial activity of GTE was significantly increased after encapsulation in nanoliposomes. The strongest antibacterial activity of nanoliposomal GTE was seen against L. monocytogenes with an inhibition zone of 16.2 mm while E. coli was the most resistance strain with an inhibition zone of 14 mm. Furthermore, the antioxidant activity of GTE was significantly increased after nanoliposome encapsulation since the IC50 value of nanoliposomal GTE was decreased to 1.78 μg/ml. Nanoencapsulation effectively enhanced beneficial properties of GTE including antimicrobial and antioxidant activities

Efficacy of vitamin E with or without probiotic, astaxanthin or rosemary extract on growth performance, survival, haematological parameters, antioxidant activity and liver enzymes in rainbow trout ( Oncorhynchus mykiss )

Growth and health status of rainbow trout (Oncorhynchus mykiss) (170.46 ± 1.16 g) fed diets containing 50 mg kg−1 vitamin E (control), 250 mg kg−1 vitamin E, 500 mg kg−1 probiotic (Bacillus lichiniformis + Bacillus subtilis each at 108 cfu g−1), 50 mg kg−1 astaxanthin and 500 mg kg−1 rosemary (Rosmarinus officinalis) extract were assessed for 4 months. No significant differences were seen in growth performance among treatments and between treatments and control (p > 0.05). The highest values of protein and lipid efficiency ratios were observed in 250 mg vitamin E, astaxanthin and rosemary trails. Probiotic trail demonstrated lower activity of liver superoxide dismutase (p 0.05). Fish fed with bacilli probiotic and rosemary extract resulted in higher survival rates compared to other treatments. Overall, these data exhibited that application of diets containing 50 mg vitamin E plus each rosemary extract (50 mg kg−1) or bacilli probiotic (500 mg kg−1) could be effective in maintaining health status of rainbow trout