Search results for: in vitro and in vivo cancer therapy

Authors: Ashleigh Williams

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Micro- and nanoplastics’ (MNLPs) omnipresence and ecological accumulation is evident when surveying recent environmental impact studies. For example, in 2014 it was estimated that at least 52.3 trillion plastic microparticles are floating at sea, and scientists have even found plastics present remote Arctic ice and snow (5,6). Plastics have even found their way into precipitation, with more than 1000 tons of microplastic rain precipitating onto the Western United States in 2020. Even more recent studies evaluating the chemical safety of reusable plastic bottles found that hundreds of chemicals leached into the control liquid in the bottle (ddH2O, ph = 7) during a 24-hour time period. A consequence of the increased abundance in plastic waste in the air, land, and water every year is the bioaccumulation of MNLPs in ecosystems and trophic niches of the animal food chain, which could potentially cause increased direct and indirect exposure of humans to MNLPs via inhalation, ingestion, and dermal contact. Though the detrimental, toxic effects of MNLPs have been established in marine biota, much less is known about the potentially hazardous health effects of chronic MNLP ingestion in humans. Recent data indicate that long-term exposure to MNLPs could cause possible inflammatory and dysbiotic effects. However, toxicity seems to be largely dose-, as well as size-dependent. In addition, the transcytotic uptake of MNLPs through the intestinal epithelia in humans remain relatively unknown. To this point, the goal of the current study was to investigate the mechanisms of micro- and nanoplastic uptake and transcytosis of Polystyrene (PE) in human stem-cell derived, physiologically relevant in vitro intestinal model systems, and to compare the relative effect of particle size (30 nm, 100 nm, 500 nm and 1 µm), and concentration (0 µg/mL, 250 µg/mL, 500 µg/mL, 1000 µg/mL) on polystyrene MNLP uptake, transcytosis and intestinal epithelial model integrity. Observational and quantitative data obtained from confocal microscopy, immunostaining, transepithelial electrical resistance (TEER) measurements, cryosectioning, and ELISA cytokine assays of the proinflammatory cytokines Interleukin-6 and Interleukin-8 were used to evaluate the localization and transcytosis of polystyrene MNPs and its impact on epithelial integrity in human-derived intestinal in vitro model systems. The effect of Microfold (M) cell induction on polystyrene micro- and nanoparticle (MNP) uptake, transcytosis, and potential inflammation was also assessed and compared to samples grown under standard conditions. Microfold (M) cells, link the human intestinal system to the immune system and are the primary cells in the epithelium responsible for sampling and transporting foreign matter of interest from the lumen of the gut to underlying immune cells. Given the uptake capabilities of Microfold cells to interact both specifically and nonspecific to abiotic and biotic materials, it was expected that M- cell induced in vitro samples would have increased binding, localization, and potentially transcytosis of Polystyrene MNLPs across the epithelial barrier. Experimental results of this study would not only help in the evaluation of the plastic toxicity, but would allow for more detailed modeling of gut inflammation and the intestinal immune system.

Keywords: nanoplastics, enteroids, intestinal barrier, tissue engineering, microfold (M) cells

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Authors: Nayer Mofidtabatabaei

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Advancements in artificial intelligence (AI) have transformed various fields, including psychology and behavioral sciences. This paper explores the diverse ways in which AI is applied to enhance research, diagnosis, therapy, and understanding of human behavior and mental health. We discuss the potential benefits and challenges associated with AI in these fields, emphasizing the ethical considerations and the need for collaboration between AI researchers and psychological and behavioral science experts. Artificial Intelligence (AI) has gained prominence in recent years, revolutionizing multiple industries, including healthcare, finance, and entertainment. One area where AI holds significant promise is the field of psychology and behavioral sciences. AI applications in this domain range from improving the accuracy of diagnosis and treatment to understanding complex human behavior patterns. This paper aims to provide an overview of the various AI applications in psychological and behavioral sciences, highlighting their potential impact, challenges, and ethical considerations. Mental Health Diagnosis AI-driven tools, such as natural language processing and sentiment analysis, can analyze large datasets of text and speech to detect signs of mental health issues. For example, chatbots and virtual therapists can provide initial assessments and support to individuals suffering from anxiety or depression. Autism Spectrum Disorder (ASD) Diagnosis AI algorithms can assist in early ASD diagnosis by analyzing video and audio recordings of children's behavior. These tools help identify subtle behavioral markers, enabling earlier intervention and treatment. Personalized Therapy AI-based therapy platforms use personalized algorithms to adapt therapeutic interventions based on an individual's progress and needs. These platforms can provide continuous support and resources for patients, making therapy more accessible and effective. Virtual Reality Therapy Virtual reality (VR) combined with AI can create immersive therapeutic environments for treating phobias, PTSD, and social anxiety. AI algorithms can adapt VR scenarios in real-time to suit the patient's progress and comfort level. Data Analysis AI aids researchers in processing vast amounts of data, including survey responses, brain imaging, and genetic information. Privacy Concerns Collecting and analyzing personal data for AI applications in psychology and behavioral sciences raise significant privacy concerns. Researchers must ensure the ethical use and protection of sensitive information. Bias and Fairness AI algorithms can inherit biases present in training data, potentially leading to biased assessments or recommendations. Efforts to mitigate bias and ensure fairness in AI applications are crucial. Transparency and Accountability AI-driven decisions in psychology and behavioral sciences should be transparent and subject to accountability. Patients and practitioners should understand how AI algorithms operate and make decisions. AI applications in psychological and behavioral sciences have the potential to transform the field by enhancing diagnosis, therapy, and research. However, these advancements come with ethical challenges that require careful consideration. Collaboration between AI researchers and psychological and behavioral science experts is essential to harness AI's full potential while upholding ethical standards and privacy protections. The future of AI in psychology and behavioral sciences holds great promise, but it must be navigated with caution and responsibility.

Keywords: artificial intelligence, psychological sciences, behavioral sciences, diagnosis and therapy, ethical considerations

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Authors: Fatima Mohammed Musa, J. B. Ameh, S. A. Ado, O. S. Olonitola

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The study was undertaken to evaluate the phytochemical constituents of extracts of Terminalia avicennioides leaf and the antidiarrhoeal effect of n-butanol fraction of the leaf extract in Swiss albino rats infected with Salmonella Typhimurium and Escherichia coli. Ethanol crude extract of Terminalia avicennioides leaf was dissolved in 1.5 liters of sterile distilled water. The extract solution was partitioned with 250 ml each of chloroform, ethyl acetate and n-butanol solvents (1:1v/v) to obtain soluble fractions from the extract. The leaf extract and its fractions were screened for the presence of phytocompounds using standard analytical methods. The antidirrhoeal activity of n-butanol fraction was evaluated in Swiss albino rats using standard methods. The results of phytochemical screening of extract of Terminalia avicennioides leaf and its fractions, revealed the presence of carbohydrates, alkaloids, tannins, flavonoids, saponins, steroids, triterpens, glycosides and phenols. The results of in vivo activity showed that 60 % of each group of rats infected with 2.0 x 108 cfu/ml viable cells of S. Typhimurium and 2.0 x109 cfu/ml viable cells of E. coli manifested the symptoms of diarrhoea, 72 hours after the rats were challenged with bacteria. Other symptoms observed among the infected animals included, loss of appetite, loss of weight, general body weakness and 40 % mortality in S. Typhimurium infected non treated group of rats. Similarly, 60 %, and 20 % mortality was observed among E. coli infected none treated and E. coli infected antibiotic (metronidazole) treated groups of rats respectively. However, there was a reduction in the number of infected rats defecating watery stools over time among all the infected rats that were treated with n-butanol fraction of the leaf extract and mortality was also not observed in the group, indicating high efficacy of n-butanol fraction of T. avicennioides leaf. The results also indicated that n-butanol can be used as alternative source of antidiarrhoeal agent in the treatment of diarrhoea caused by Salmonella Typhimurium and Escherichia coli. In the light of this, there is a need for further research on the mechanism of action of the candidate fraction of T. avicennioides leaf which could be responsible for the observed in vivo antibacterial activity.

Keywords: antidirrhoeal effect, phytochemical constituents, swiss albino rats, terminalia avicennioides

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Authors: Amir Amiri, Naghmeh Morakabati

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During the recent years the six-fold growth of cancer in Iran has led the production of healthy products to become a challenge in the food industry. Due to the young population in the country, the consumption of fast foods is growing. The chemical cancer-causing preservatives are used to produce these products more than the standard; so using an appropriate alternative seems to be important. On the one hand, the plant essential oils show the high antimicrobial potential against pathogenic and spoilage microorganisms and on the other hand they are highly volatile and decomposed under the processing conditions. The study aims to produce the loaded chitosan nanoparticles with different concentrations of savory essential oil to improve the anti-microbial property and increase the resistance of essential oil to oxygen and heat. The encapsulation efficiency was obtained in the range of 32.07% to 39.93% and the particle size distribution of the samples was observed in the range of 159 to 210 nm. The range of Zeta potential was obtained between -11.9 to -23.1 mV. The essential oil loaded in chitosan showed stronger antifungal activity against Rhizopus stolonifer. The results showed that the antioxidant property is directly related to the concentration of loaded essential oil so that the antioxidant property increases by increasing the concentration of essential oil. In general, it seems that the savory essential oil loaded in chitosan particles can be used as a food processor.

Keywords: chitosan, encapsulation, essential oil, nanogel

Procedia PDF Downloads 257

Authors: Yasmin A. M. Ferreira, Ana C. K. Pelissari, Sofia De C. F. Vicente, Raquel M. Da S. Campos, Deborah C. L. Masquio, Lian Tock, Lila M. Oyama, Flavia C. Corgosinho, Valter T. Boldarine, Ana R. Dâmaso

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The prevalence of overweight and obesity is growing around the world and currently considered a global epidemic. Food and nutrition are essential requirements for promoting health and protecting non-communicable chronic diseases, such as obesity and cardiovascular disease. Specific dietary components may modulate the inflammation and oxidative stress in obese individuals. Few studies have investigated the dietary Inflammation Factor Rating (IFR) in obese adolescents. The IFR was developed to characterize an individual´s diet on anti- to pro-inflammatory score. This evaluation contributes to investigate the effects of inflammatory diet in metabolic profile in several individual conditions. Objectives: The present study aims to investigate the effects of a multidisciplinary weight loss therapy on inflammation factor rating and cardiometabolic risk in obese adolescents. Methods: A total of 26 volunteers (14-19 y.o) were recruited and submitted to 20 weeks interdisciplinary therapy allied to health education website- Ciclo do Emagrecimento®, including clinical, nutritional, psychological counseling and exercise training. The body weight was monitored weekly by self-report and photo. The adolescents answered a test to evaluate the knowledge of the topics covered in the videos. A 24h dietary record was applied at the baseline and after 20 weeks to assess the food intake and to calculate IFR. A negative IFR suggests that diet may have inflammatory effects and a positive IFR indicates an anti-inflammatory effect. Statistical analysis was performed using the program STATISTICA version 12.5 for Windows. The adopted significant value was α ≤ 5 %. Data normality was verified with the Kolmogorov Smirnov test. Data were expressed as mean±SD values. To analyze the effects of intervention it was applied test t. Pearson´s correlations test was performed. Results: After 20 weeks of treatment, body mass index (BMI), body weight, body fat (kg and %), abdominal and waist circumferences decreased significantly. The mean of high-density lipoprotein cholesterol (HDL-c) increased after the therapy. Moreover, it was found an improvement of inflammation factor rating from -427,27±322,47 to -297,15±240,01, suggesting beneficial effects of nutritional counselling. Considering the correlations analysis, it was found that pro-inflammatory diet is associated with increase in the BMI, very low-density lipoprotein cholesterol (VLDL), triglycerides, insulin and insulin resistance index (HOMA-IR); while an anti-inflammatory diet is associated with improvement of HDL-c and insulin sensitivity Check index (QUICKI). Conclusion: The 20-week blended multidisciplinary therapy was effective to reduce body weight, anthropometric circumferences and improve inflammatory markers in obese adolescents. In addition, our results showed that an increase in inflammatory profile diet is associated with cardiometabolic parameters, suggesting the relevance to stimulate anti-inflammatory diet habits as an effective strategy to treat and control of obesity and related comorbidities. Financial Support: FAPESP (2017/07372-1) and CNPq (409943/2016-9)

Keywords: cardiometabolic risk, inflammatory diet, multidisciplinary therapy, obesity

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Authors: Anne Giles

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Humans naturally wish they could forget traumatic experiences. To help prevent future harm, however, the human brain has evolved to retain data about experiences of threat, alarm, or violation. When given compassionate support and assistance with thinking helpfully and realistically about traumatic events, most people can adjust to experiencing hardships, albeit with residual sad, unfortunate memories. Persistent, recurrent, intrusive memories, difficulty sleeping, emotion dysregulation, and avoidance of reminders, however, may be symptoms of Post-traumatic Stress Disorder (PTSD). Brain scans show that PTSD affects brain functioning. We currently have no physical means of restoring the system of brain structures and functions involved with PTSD. Medications may ease some symptoms but not others. However, forms of "talk therapy" with cognitive components have been found by researchers to reduce, even resolve, a broad spectrum of trauma symptoms. Many cultures have taboos against talking about hardships. Individuals may present themselves to mental health care professionals with severe, disabling trauma symptoms but, because of cultural norms, be unable to speak about them. In China, for example, relationship expectations may include the belief, "Bad things happening in the family should stay in the family (jiāchǒu bùkě wàiyán 家丑不可外扬)." The concept of "family (jiā 家)" may include partnerships, close and extended families, communities, companies, and the nation itself. In contrast to many trauma therapies, Cognitive Processing Therapy (CPT) for Post-traumatic Stress Disorder asks its participants to focus not on "what" happened but on "why" they think the trauma(s) occurred. The question "why" activates and exercises cognitive functioning. Brain scans of individuals with PTSD reveal executive functioning portions of the brain inadequately active, with emotion centers overly active. CPT conceptualizes PTSD as a network of cognitive distortions that keep an individual "stuck" in this under-functioning and over-functioning dynamic. Through asking participants forms of the question "why," plus offering a protocol for examining answers and relinquishing unhelpful beliefs, CPT assists individuals in consciously reactivating the cognitive, executive functions of their brains, thus restoring normal functioning and reducing distressing trauma symptoms. The culturally sensitive components of CPT that allow people to "talk about it without talking about it" may offer the possibility for worldwide relief from symptoms of trauma.

Keywords: cognitive processing therapy (CPT), cultural norms, post-traumatic stress disorder (PTSD), trauma recovery

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Authors: Jae Bok Heo, Yun Mi Lee, Hee Rang Yun

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Several GTPases are required for ribosome biogenesis and assembly. We recently characterized rice (Oryza sativa) nuclear/nucleolar GTPase 2 (OsNug2), belonging to the YlqF/YawG family of GTPases, as playing a role in pre-60S ribosomal subunit maturation. To investigate the potential factors involved in regulating the function of OsNug2, yeast two-hybrid screens were carried out using OsNug2 as bait. Rice serine/threonine kinase 1 (OsSTK1) was identified as a potential interacting protein candidate. In vitro pull down and bimolecular fluorescence complementation assays confirmed the interaction between OsNug2 and OsSTK1, and like green fluorescent protein-tagged OsNug2, green fluorescent protein-tagged OsSTK1 was targeted to the nucleus of Arabidopsis protoplasts. OsSTK1 was not found to affect the GTP-binding activity of OsNug2; however, when recombinant OsSTK1 was included in OsNug2 assay reaction mixtures, OsSTK1 increased the GTPase activity of OsNug2. To test whether OsSTK1 phosphorylates OsNug2 in vitro, a kinase assay was performed. OsSTK1 was found to have weak autophosphorylation activity and strongly phosphorylated serine 209 of OsNug2. Yeast complementation testing resulted in a GAL::OsNug2(S209N) mutant-harboring yeast strain exhibiting a growth-defective phenotype on galactose medium at 39°C, divergent from that of a yeast strain harboring GAL::OsNug2. The intrinsic GTPase activity of mutant OsNug2(S209N) was found to be similar to that of OsNug2, was not fully enhanced upon weak binding of OsSTK1. Our findings reported here indicate that OsSTK1 functions as a positive regulator protein of OsNug2 by enhancing the GTPase activity of OsNug2, and that the phosphorylation of serine 209 of OsNug2 is essential for the complete function of OsNug2 in ribosome biogenesis.

Keywords: OsSTK1, OsNug2, GTPase activity, GTP binding activity, phosphorylation

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Authors: R. Razavizadeh, F. Adabavazeh, M. Rezaee Chermahini

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Salinity is one of the most widespread agricultural problems in arid and semi-arid areas that limits the plant growth and crop productivity. In this study, the salt stress effects on protein, reducing sugar, proline contents and antioxidant enzymes activities of Carum copticum L. under in vitro conditions were studied. Seeds of C. copticum were cultured in Murashige and Skoog (MS) medium containing 0, 25, 50, 100 and 150 mM NaCl and calli were cultured in MS medium containing 1 μM 2, 4-dichlorophenoxyacetic acid, 4 μM benzyl amino purine and different levels of NaCl (0, 25, 50, 100 and 150 mM). After NaCl treatment for 28 days, the proline and reducing sugar contents of shoots, roots and calli increased significantly in relation to the severity of the salt stress. The highest amount of proline and carbohydrate were observed at 150 and 100 mM NaCl, respectively. The reducing sugar accumulation in shoots was the highest as compared to roots, whereas, proline contents did not show any significant difference in roots and shoots under salt stress. The results showed significant reduction of protein contents in seedlings and calli. Based on these results, proteins extracted from the shoots, roots and calli of C. copticum treated with 150 mM NaCl showed the lowest contents. The positive relationships were observed between activity of antioxidant enzymes and the increase in stress levels. Catalase, ascorbate peroxidase and superoxide dismutase activity increased significantly under salt concentrations in comparison to the control. These results suggest that the accumulation of proline and sugars, and activation of antioxidant enzymes play adaptive roles in the adaptation of seedlings and callus of C. copticum to saline conditions.

Keywords: antioxidant enzymes, Carum copticum, organic solutes, salt stress

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Authors: Mitesh Rathod, Jungho Ahn, Noo Li Jeon, Junghoon Lee

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Endothelial cells respond to cues from both biochemical as well as micro mechanical environment. Significant effort has been directed to understand the effects of biochemical signaling, however, relatively little is known about regulation of endothelial cell biology by the micro mechanical environment. Numerous studies have been performed to understand how physical forces regulate endothelial cell behavior. In this regard, past studies have majorly focused on exploring how fluid shear stress governs endothelial cell behavior. Parallel plate flow chambers and rectangular microchannels are routinely employed for applying fluid shear force on endothelial cells. However, these studies fall short in mimicking the in vivo like micro environment from topological aspects. Few studies have only used circular microchannels to replicate in vivo like condition. Seldom efforts have been directed to elucidate the combined effect of topology, substrate rigidity and fluid shear stress on endothelial cell response. In this regard, we demonstrate a facile fabrication process to develop a hybrid polydimethylsiloxane microfluidic platform to study endothelial cell biology. On a single chip microchannels with different cross sections i.e., circular, rectangular and square have been fabricated. In addition, our fabrication approach allows variation in the substrate rigidity along the channel length. Two different variants of polydimethylsiloxane, namely Sylgard 184 and Sylgard 527, were utilized to achieve the variation in rigidity. Moreover, our approach also enables in creating Y bifurcation circular microchannels. Our microfluidic platform thus facilitates for conducting studies pertaining to endothelial cell morphology with respect to change in topology, substrate rigidity and fluid flow on a single chip. The hybrid platform was tested by culturing Human Umbilical Vein Endothelial Cells in circular microchannels with varying substrate rigidity, and exposed to fluid shear stress of 12 dynes/cm² and static conditions. Results indicate the cell area response to flow induced shear stress was governed by the underlying substrate mechanics.

Keywords: hybrid, microfluidic platform, PDMS, shear flow, substrate rigidity

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Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: trichoderma spp., meloidogyne javanica, Fusarium oxysporum f.sp.radicis lycopersici, biocontrol

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Authors: Latife Merve Oktay, Berrin Tugrul

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Hydroxytyrosol (HT) is a phenolic phytochemical molecule derived from the hydrolysis of oleuropein, which originates during the maturation of the olives. It has recently received particular attention because of its antioxidant, anti-proliferative, pro-apoptotic and anti-inflammatory activities. In this study, we investigated the cytotoxic and apoptotic effects of hydroxytyrosol and its effects on phosphatidylinositol 3-kinase/Akt (PI3K/Akt) and extracellular signal-regulated kinase 1/2 (ERK 1/2) signaling pathways in human ovarian cancer cell lines OVCAR-3 and MDAH-2774. XTT cell proliferation kit, Cell Death Detection Elisa Plus Kit (Roche) and Human Apoptosis Array (R&D Systems) were used to determine the cytotoxic and apoptotic effects of HT in OVCAR-3 and MDAH-2774 cell lines at 24, 48, 72, and 96 h. Effect of HT on PI3K/Akt and ERK 1/2 signaling pathways were investigated by using specific inhibitors of these pathways. IC50 values of HT were found to be 102.3 µM in MDAH-2774 cells at 72 h and 51.5 µM in OVCAR-3 cells at 96 h. Apoptotic effect of HT in MDAH-2774 cells was the highest at 50 µM at 72 h, and kept decreasing at 100 and 150 µM concentrations and was not seen at 200 µM and higher concentrations. Highest apoptotic effect was seen at 100 µM concentration in OVCAR-3 cells at 96 h, however apoptotic effect was decreased over 100 µM concentrations. According to antibody microarray results, HT increased the levels of pro-apoptotic molecules Bad, Bax, active caspase-3, Htra2/Omi by 2.0-, 1.4-, 1.2-, 4.2-fold, respectively and also increased the levels of pro-apoptotic death receptors TRAIL R1/DR4, TRAIL R2/DR5, FAS/TNFRSF6 by 2.1-, 1.7-, 1.6-fold, respectively, however, it decreased the level of Survivin by 1.6-fold which is one of the inhibitor of apoptosis protein (IAP) family in MDAH-2774 cells. In OVCAR-3 cells, HT decreased the levels of anti-apoptotic proteins Bcl-2, pro-caspase 3 by 3.1-, 8.2-fold, respectively and IAP family proteins CIAP-1, CIAP-2, XIAP, Livin, Survivin by 6.5-, 6.0-, 3.2-, 2.2-, 2.7-fold, respectively and increased the level of cytochrome-c by 1.2-fold. We have shown that HT shows its cytotoxic and apoptotic effect through inhibiting ERK 1/2 signaling pathway in both OVCAR-3 and MDAH-2774 cells. Further studies are needed to investigate molecular mechanisms and modulatory effects of hydroxytyrosol.

Keywords: apoptosis, cytotoxicity, hydroxytyrosol, ovarian cancer

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Authors: Sandra Soca Lozano, Teresa Isabel Lozano Pérez, Germain Weber

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Cancer is a chronic disease of high prevalence in children and adolescents. Medical care in Cuba is carried out by a multidisciplinary team and family is the mediator between this team and the patient. Around this disease, there are interwoven many stereotypes and taboos by its relation to death. In this research report, we describe the work paradigm of psychological care to patients suffering from these diseases in the University Pediatric Hospital Juan Manuel Márquez of Havana, Cuba. We present the psychosocial factors that must be taken into account to provide comprehensive care and ensuring the quality of life of patients and their families. We also present the factors related to the health team and the management of information done with the patient. This is a descriptive proposal from the working experience accumulated in the named institution and in the review of the literature. As a result of this report we make a proposal of teamwork and the aspects in which psychological intervention should be continue performing in terms of increasing the quality of the care made by the health team. We conclude that it is necessary to continue improving the information management of children and adolescents with theses health problems and took into account their right to autonomy.

Keywords: comprehensive care, management of information, psychosocial factors, right to autonomy

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Authors: Doni Dermawan

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Modern sports competition as a consequence of the increase in the value of the business and entertainment in the field of sport has been demanding athletes to always have excellent physical endurance performance. Physical exercise is done in a long time, and intensive may pose a risk of muscle tissue damage caused by the increase of the enzyme creatine kinase. Branched Chain Amino Acids (BCAA) is an essential amino acid that is composed of leucine, isoleucine, and valine which serves to maintain muscle tissue, keeping the immune system, and prevent further loss of coordination and muscle pain. Pea (Pisum sativum L.) is a kind of leguminous plants that are rich in Branched Chain Amino Acids (BCAA) where every one gram of protein pea contains 82.7 mg of leucine; 56.3 mg isoleucine; and 56.0 mg of valine. This research aims to develop Branched Chain Amino Acids (BCAA) from pea extract is applied in dosage forms Gel PVP Kinesio Tape technology using Ultrasound-assisted Extraction. The method used in the writing of this paper is the Cochrane Collaboration Review that includes literature studies, testing the quality of the study, the characteristics of the data collection, analysis, interpretation of results, and clinical trials as well as recommendations for further research. Extraction of BCAA in pea done using ultrasound-assisted extraction technology with optimization variables includes the type of solvent extraction (NaOH 0.1%), temperature (20-250C), time (15-30 minutes) power (80 watt) and ultrasonic frequency (35 KHz). The advantages of this extraction method are the level of penetration of the solvent into the membrane of the cell is high and can increase the transfer period so that the BCAA substance separation process more efficient. BCAA extraction results are then applied to the polymer PVP (Polyvinylpyrrolidone) Gel powder composed of PVP K30 and K100 HPMC dissolved in 10 mL of water-methanol (1: 1) v / v. Preparations Kinesio Tape Gel PVP is the BCAA in the gel are absorbed into the muscle tissue, and joints through tensile force then provides stimulation to the muscle circulation with variable pressure so that the muscle can increase the biomechanical movement and prevent damage to the muscle enzyme creatine kinase. Analysis and evaluation of test preparation include interaction, thickness, weight uniformity, humidity, water vapor permeability, the levels of the active substance, content uniformity, percentage elongation, stability testing, release profile, permeation in vitro and in vivo skin irritation testing.

Keywords: branched chain amino acid, BCAA, Kinesio tape, pea, PVP gel, ultrasound-assisted extraction

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Authors: James Patrick A. Diaz, Raul E. Ramboyong

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Background: Hypertensive response during maximal exercise test provides important information on the level of blood pressure control and evaluation of treatment. Method: A single center retrospective descriptive study was conducted among 117 young (aged 20 to 40) and middle age (aged 40 to 65) hypertensive patients, who underwent treadmill stress test. Currently on maintenance frontline medication either monotherapy (Angiotensin-converting enzyme inhibitor/Angiotensin receptor blocker [ACEi/ARB], Calcium channel blocker [CCB], Diuretic - Hydrochlorthiazide [HCTZ]) or combination therapy (ARB+CCB, ARB+HCTZ), who attained a maximal exercise on treadmill stress test (TMST) with hypertensive response (systolic blood pressure: male >210 mm Hg, female >190 mm Hg, diastolic blood pressure >100 mmHg, or increase of >10 mm Hg at any time during the test), on Bruce and Modified Bruce protocol. Exaggerated blood pressure response during exercise (systolic [SBP] and diastolic [DBP]), peak exercise blood pressure (SBP and DBP), recovery period (SBP and DBP) and test for ischemia and their antihypertensive medication/s were investigated. Analysis of variance and chi-square test were used for statistical analysis. Results: Hypertensive responses on maximal exercise test were seen mostly among female population (P < 0.000) and middle age (P < 0.000) patients. Exaggerated diastolic blood pressure responses were significantly lower in patients who were taking CCB (P < 0.004). A longer recovery period that showed a delayed decline in SBP was observed in patients taking ARB+HCTZ (P < 0.036). There were no significant differences in the level of exaggerated systolic blood pressure response and during peak exercise (both systolic and diastolic) in patients using either monotherapy or combination antihypertensives. Conclusion: Calcium channel blockers provided lower exaggerated diastolic BP response during maximal exercise test in hypertensive middle age patients. Patients on combination therapy using ARB+HCTZ exhibited a longer recovery period of systolic blood pressure.

Keywords: antihypertensive, exercise test, hypertension, hyperytensive response

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Authors: Hanan Mohamed Abd Elmoneim, Rawan Saleh AlJawi, Razan Saleh AlJawi, Aseel Abdullah AlMasoudi , Zyad Adnan Turkistani, Anas Abdulkarim Alkhoutani , Ohood Musaed AlJuhani , Hanan Attiyah AlZahrani

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Background Esophageal cancer is the eighth most common cancer worldwide. More than 90% of esophageal cancers are either squamous cell carcinoma or adenocarcinoma. Squamous dysplasia is a precancerous lesion for squamous cell carcinoma and Barrett's esophagus is the precancerous lesion for adenocarcinoma. Gastro-esophageal reflux disease (GERD) is the initiation factor for Barrett's esophagus. Cyclooxygenase-2 (COX-2) is a key enzyme in arachidonic metabolism. It appears to play an important role in gastrointestinal carcinogenesis. COX-2 activity may be a potential target for the prevention of cancer progression by selective COX-2 inhibitors, which decrease proliferation and increase apoptosis. Objectives To assess COX-2 expression in premalignant and malignant esophageal epitheliums changes and detect its roles in progression of these lesions. Materials and Methods We analyzed the expression of COX-2 immunohistochemically in 40 esophageal biopsies utilizing the streptavidin-biotin-peroxidase complex method on archival formalin fixed-paraffin embedded blocks. Histopathologically, 17 (42.5%) of cases were non-malignant cases which included GERD, Barrett's esophagus and squamous dysplasia. The malignant cases were 23 (57.5%) squamous cell carcinoma, adenocarcinoma and undifferentiated carcinoma. Results In non-malignant cases 7 (41.2%) out of 17 cases had high COX-2 expression. In squamous cell carcinoma 10 (83.3%) out of 12 cases had high COX-2 expression. The expression of COX-2 was high in all 9 (100%) cases of adenocarcinoma. COX-2 expression is significantly increased (P=0.005 and P=0.0001) in squamous cell carcinoma and adenocarcinoma respectively. There was a significant difference in COX-2 immunoreactivity between malignant and non-malignant lesions (P=0.0003). Conclusion COX-2 is responsible for the progression of esophageal diseases from benign to malignant. We recommend that COX-2 immunohistochemistry should be done routinely for premalignant and malignant esophageal lesions as selective COX-2 inhibitors will be helpful in the treatment. Further studies on molecular and genetic basis of COX-2 expression are needed to unmask its role and relation to progression of esophageal lesions.

Keywords: Cox-2, Esophageal adinocarcinoma, Esophageal squamous cell carcinoma, Immunohistochemistry.

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Authors: N. Selvi Gunel, L. M. Oktay, H. Memmedov, B. Durmaz, H. Kalkan Yildirim, E. Yildirim Sozmen

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Object: Propolis which consists of compounds that are accepted as antioxidant, antimicrobial, antiseptic, antibacterial, anti-inflammatory, anti-mutagenic, immune-modulator and cytotoxic, is frequently used in current therapeutic applications. However, some of them result in allergic side effects, causing consumption to be restricted. Previously our group has succeeded in producing a new biotechnological product which was less allergenic. In this study, we purpose to optimize production conditions of this biologically-transformed propolis and determine the cytotoxic effects of obtained new products on colon cancer cell line (HCT-116). Method: Firstly, solid propolis samples were dissolved in water after weighing, grinding and sizing (sieve-35mesh) and applied 40 kHz/10 min ultrasonication. Samples were prepared according to inoculation with Lactobacillus plantarum in two different proportions (2.5% and 3.5%). Chromatographic analyzes of propolis were performed by UPLC-MS/MS (Waters, Milford, MA) system. Results were analysed by UPLC-MS/MS system MassLynx™ 4.1 software. HCT-116 cells were treated with propolis examples at 25-1000 µg/ml concentrations and cytotoxicity were measured by using WST-8 assay at 24, 48, and 72 hours. Samples with biological transformation were compared with the non-transformed control group samples. Our experiment groups were formed as follows: untreated (group 1), propolis dissolved in water ultrasonicated at 40 kHz/10 min (group 2), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 2.5% L. plantarum L1 strain (group 3), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 3.5% L. plantarum L3 strain (group 4). Obtained data were calculated with Graphpad Software V5 and analyzed by two-way ANOVA test followed by Bonferroni test. Result: As a result of our study, the cytotoxic effect of propolis samples on HCT-116 cells was evaluated. There was a 7.21 fold increase in group 3 compared to group 2 in the concentration of 1000 µg/ml, and it was a 6.66 fold increase in group 3 compared to group 1 at the end of 24 hours. At the end of 48 hours, in the concentration of 500 µg/ml, it was determined 4.7 fold increase in group 4 compared to group 3. At the same time, in the concentration of 750 µg/ml it was determined 2.01 fold increase in group 4 compared to group 3 and in the same concentration, it was determined 3.1 fold increase in group 4 compared to group 2. Also, at the 72 hours, in the concentration of 750 µg/ml, it was determined 2.42 fold increase in group 3 according to group 2 and in the same time, in the concentration of 1000 µg/ml, it was determined 2.13 fold increase in group 4 according to group 2. According to cytotoxicity results, the group which were ultrasonicated at 40 kHz/10min and inoculated 3.5% L. plantarum L3-strain had a higher cytotoxic effect. Conclusion: It is known that bioavailability of propolis is halved in six months. The data obtained from our results indicated that biologically-transformed propolis had more cytotoxic effect than non-transformed group on colon cancer cells. Consequently, we suggested that L. plantarum-transformation provides both reduction of allergenicity and extension of bioavailability period by enhancing healthful polyphenols.

Keywords: bio-transformation, propolis, colon cancer, cytotoxicity

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Authors: T. Ferreira, A. Kulkarni, C. Bretscher, K. Richter, M. Ehrlich, A. Marchini

Abstract:

H-1 protoparvovirus (H-1PV) is a virus with inherent oncolytic and oncosuppressive activities while remaining non-pathogenic in humans. H-1PV was the first oncolytic parvovirus to undergo clinical testing. Results from trials in patients with glioblastoma or pancreatic carcinoma showed an excellent safety profile and first signs of efficacy. H-1PV infection is vastly dependent on cellular factors, from cell attachment and entry to viral replication and egress. Hence, we believe that the characterisation of the parvovirus life cycle would ultimately help further improve H-1PV clinical outcome. In the present study, we explored the entry pathway of H-1PV in cervical HeLa and glioma NCH125 cancer cell lines. Electron and confocal microscopy showed viral particles associated with clathrin-coated pits and vesicles, providing the first evidence that H-1PV cell entry occurs through clathrin-mediated endocytosis. Accordingly, we observed that by blocking clathrin-mediated endocytosis with hypertonic sucrose, chlorpromazine, or pitstop 2, H-1PV transduction was markedly decreased. Accordingly, siRNA-mediated knockdown of AP2M1, which retains a crucial role in clathrin-mediated endocytosis, verified the reliance of H-1PV on this route to enter HeLa and NCH125 cancer cells. By contrast, we found no evidence of viral entry through caveolae-mediated endocytosis. Indeed, pre-treatment of cells with nystatin or methyl-β-cyclodextrin, both inhibitors of caveolae-mediated endocytosis, did not affect viral transduction levels. Unexpectedly, siRNA-mediated knockdown of caveolin-1, the main driver of caveolae-mediated endocytosis, increased H-1PV transduction, suggesting caveolin-1 is a negative modulator of H-1PV infection. We also show that H-1PV entry is dependent on dynamin, a protein responsible for mediating the scission of vesicle neck and promoting further internalisation. Furthermore, since dynamin inhibition almost completely abolished H-1PV infection, makes it unlikely that H-1PV uses macropinocytosis as an alternative pathway to enter cells. After viral internalisation, H-1PV passes through early to late endosomes as observed by confocal microscopy. Inside these endocytic compartments, the acidic environment proved to be crucial for a productive infection. Inhibition of acidification of pH dramatically reduced H-1PV transduction. Besides, a fraction of H-1PV particles was observed inside LAMP1-positive lysosomes, most likely following a non-infectious route. To the author's best knowledge, this is the first study to characterise the cell entry pathways of H-1PV. Along these lines, this work will further contribute to understand H-1PV oncolytic properties as well as to improve its clinical potential in cancer virotherapy.

Keywords: clathrin-mediated endocytosis, H-1 parvovirus, oncolytic virus, virus entry

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Authors: Nanteza Rachel

Abstract:

Background: Worldwide, there were 36.7 million people living with Human Immunodeficiency Virus (HIV) in 2015, up from 35 million at the end of 2013. Wakiso district is one of the hotspots for the Human Immunodeficiency Virus (HIV)/ Acquired Immune Deficiency Syndrome (AIDS) infection in Uganda, with the prevalence of 8.1 %. Herbal medicine has gained popularity among Human Immunodeficiency Virus (HIV)/ Acquired Immune Deficiency Syndrome (AIDS) patients as adjuvant therapy to reduce the adverse effects of ART. Regardless of the subsidized and physical availability of the Anti-Retroviral Therapy (ART), majority of Africans living with Human Immunodeficiency Virus (HIV)/ Acquired Immune Deficiency Syndrome (AIDS) resort to adding to their ART traditional medicine. Result found out from a pilot observation made by the PI that indicate 13 out of 30 People Living with AIDS(PLWA) who are attending Human Immunodeficiency Virus (HIV) clinics in Wakiso district reported to be using herbal preparations despite the fact that they were taking Anti Retro Viral (ARVs) this prompted this study to be done. Purpose of the study: To determine the prevalence and factors associated with concurrent use of herbal medicine and anti-retroviral therapy among HIV/AIDS patients attending selected HIV clinics in Wakiso district. Methodology: This was a cross sectional study with both quantitative data collection (use of a questionnaire) and qualitative data collection (key informants’ interviews). A mixed method of sampling was used, that is, purposive and random sampling. Purposive sampling was based on the location in the district and used to select 7 health facilities basing on the 7 health sub districts from Wakiso. Simple random sampling was used to select one HIV clinic from each of the 7 health sub districts. Furthermore, the study units were enrolled in to the study as they entered into the HIV clinics, and 105 respondents were interviewed. Both manual and computer packages (SPSS) were used to analyze the data Results: The prevalence of concurrent use of herbal medicine and ART was 38 (36.2%). Commonly HIV symptom treated with herbs was fever 27(71.1%), diarrhea 3(7.9%) and cough 2(5.3%). Commonly used herbs for fever (Omululuza (Vernonica amydalina), Ekigagi (Aloe sp), Nalongo (Justicia betonica Linn) while for diarrhea was Ntwatwa. The side effects also included; too much pain, itchy pain of HIV, aneamia,felt sick, loss/gain appetite, joint pain and bad dreams. Herbs used to sooth the side effects were; for aneamia was avocado leaves Parea Americana mill The significant factors associated with concurrent use of herbal medicine were being familiar with herbs and conventional medicine for management HIV symptoms being expensive. The other significant factor was exhibiting hostility to patients by health personnel providing HIV care. Conclusion: Herbal medicine is widely used by clients in HIV/AIDS care. Patients being familiar with herbs and conventional medicine being expensive were associated with concurrent use of herbal medicine and ART. The exhibition of hostility to the HIV/AIDS patients by the health care providers was also associated with concurrent use of herbal medicine and ART among HIV/AIDS patients.

Keywords: HIV patients, herbal medicine, antiretroviral therapy, factors associated

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Authors: Georgia Saxami, Evangelia N. Kerezoudi, Evdokia K. Mitsou, Marigoula Vlassopoulou, Georgios Zervakis, Adamantini Kyriacou

Abstract:

Autism Spectrum Disorder (ASD) is a complex group of developmental disorders of the brain, characterized by social and communication dysfunctions, stereotypes and repetitive behaviors. The potential interaction between gut microbiota (GM) and autism has not been fully elucidated. Children with autism often suffer gastrointestinal dysfunctions, while alterations or dysbiosis of GM have also been observed. Treatment with dietary components has been postulated to regulate GM and improve gastrointestinal symptoms, but there is a lack of evidence for such approaches in autism, especially for prebiotics. This study assessed the effects of Pleurotus eryngii mushroom (candidate prebiotic) and inulin (known prebiotic compound) on gut microbial composition, using faecal samples from autistic children in an in vitro batch culture fermentation system. Selected members of GM were enumerated at baseline (0 h) and after 24 h fermentation by quantitative PCR. After 24 h fermentation, inulin and P. eryngii mushroom induced a significant increase in total bacteria and Faecalibacterium prausnitzii compared to the negative control (gut microbiota of each autistic donor with no carbohydrate source), whereas both treatments induced a significant increase in levels of total bacteria, Bifidobacterium spp. and Prevotella spp. compared to baseline (t=0h) (p for all <0.05). Furthermore, this study evaluated the impact of fermentation supernatants (FSs), derived from P. eryngii mushroom or inulin, on the expression levels of tight junctions’ genes (zonulin-1, occludin and claudin-1) in Caco-2 cells stimulated by bacterial lipopolysaccharides (LPS). Pre-incubation of Caco-2 cells with FS from P. eryngii mushroom led to a significant increase in the expression levels of zonulin-1, occludin and claudin-1 genes compared to the untreated cells, the cells that were subjected to LPS and the cells that were challenged with FS from negative control (p for all <0.05). In addition, incubation with FS from P. eryngii mushroom led to the highest mean expression values for zonulin-1 and claudin-1 genes, which differed significantly compared to inulin (p for all <0.05). Overall, this research highlighted the beneficial in vitro effects of P. eryngii mushroom on the composition of GM of autistic children after 24 h of fermentation. Also, our data highlighted the potential preventive effect of P. eryngii FSs against dysregulation of the intestinal barrier, through upregulation of tight junctions’ genes associated with the integrity and function of the intestinal barrier. This research has been financed by "Supporting Researchers with Emphasis on Young Researchers - Round B", Operational Program "Human Resource Development, Education and Lifelong Learning."

Keywords: gut microbiota, intestinal barrier, autism spectrum disorders, Pleurotus Eryngii

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Authors: Nadeem H. Meikha , Nazad H. Qader, Basheer M. Hasafa

Abstract:

The trial was conducted to examine the effect of He-Ne laser therapy on the testis and serum testosterone level in adult rats. Thirty five albino Western adult male rats aged 3-4 months and weighing approximately 250-300 g were used and divided into three treatments. Testicular tissue of rats in the first and second treatments were exposed once daily for three successively days to a dose of irradiation 1.02 j/cm2 (40 second), and to 2.03 j/cm2 (80 second) respectively, while the third group left without any treatments (control). The results showed that the process of irradiation adversely affected on the level of serum testosterone concentration of the irradiated rats in the first and second treatment comparing to the normal level in the control group. While the histological examination showed that decrease in number of germ cells with 40 second of irradiation at day three, with 80 second of irradiation the decreased started at day two and three. The spermatids number decreased in rate low, medium, high respectively for three days of 40 second of irradiation, while the spermatids number were adversely affected by dropping in a rate of medium, large and very large for three days of 80 second of irradiation, respectively. In conclusion our study revealed that any reduction in sertoli cells causes adverse affect on both spermatids and germinal cells which increase with the increasing of duration and repetition of irradiation.

Keywords: He-Ne laser, rats, testosterone, spermatids

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Authors: Asma Chbani, Douaa Salim, Josephine Al Alam, Pascale De Caro

Abstract:

Penicillium digitatum, the causal agent of citrus green mold, is responsible for 90% of post-harvest losses. Chemical fungicides remain the most used products for protection against this pathogen but are also responsible for damage to human health and the environment. The aim of this study is to evaluate the ability of Spinacia oleracea extract to serve as biological control agents, an alternative to harmful synthetic fungicides, against orange decay for storing fruit caused by P. digitatum. In this study, we studied the implication of a crude extract of a green plant, Spinacia oleracea, in the protection of oranges against P. digitatum. Thus, in vivo antifungal tests as well as adhesion test were done. For in vivo antifungal test, oranges were pulverized with the prepared crude extracts at different concentrations ranged from 25 g L⁻¹ to 200 g L⁻¹, contaminated by the fungus and then observed during 8 weeks for their macroscopic changes at 24°C. For adhesion test, the adhesion index is defined as the number of Penicillium digitatum spores fixed per orange cell. An index greater than 25 is the indicator of a strong adhesion, whereas for an index less than 10, the adhesion is low. Ten orange cells were examined in triplicate for each extract, and the averages of adherent cells were calculated. Obtained results showed an inhibitory activity of the Penicillium development with the aqueous extract of dry Spinacia oleracea with a concentration of 50 g L⁻¹ considered as the minimal protective concentration. The prepared extracts showed a greater inhibition of the development of P. digitatum up to 10 weeks, even greater than the fungicide control Nystatin. Adhesion test’s results showed that the adhesion of P. digitatum spores to the epidermal cells of oranges in the presence of the crude spinach leaves extract is weak; the mean of the obtained adhesion index was estimated to 2.7. However, a high adhesion was observed with water used a negative control. In conclusion, all these results confirm that the use of this green plant highly rich in chlorophyll having several phytotherapeutic activities, could be employed as a great treatment for protection of oranges against mold and also as an alternative for chemical fungicides.

Keywords: Penicillium digitatum, Spinacia oleracea, oranges, biological control, postharvest diseases

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Authors: S. Beekrum, B. Odhav, R. Lalloo, E. A. Amonsou

Abstract:

Marine microalgae are rich sources of novel and biologically active metabolites; therefore they may be used in the food industry as natural food ingredients and functional foods. They have several biological applications related to health benefits, among others. The aim of the study focused on the screening of phytochemicals from Amphora sp. biomass extracts, and to examine the in vitro antioxidant and antimicrobial potential. Amphora sp. biomass was obtained from CSIR (South Africa) and methanol, hexane and water extracts were prepared. The in vitro antimicrobial effect of extracts were tested against some pathogens (Staphylococcus aureus, Listeria monocytogenes, Bacillus subtilis, Salmonella enteritidis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans), using the disc diffusion assay. Qualitative analyses of phytochemicals were conducted by chemical tests. The present investigation revealed that all extracts showed relatively strong antibacterial activity against most of the tested bacteria. The highest phenolic content was found in the methanolic extract. Results of the DPPH assay showed that the biomass contained strong antioxidant capacity, 79% in the methanolic extract and 85% in the hexane extract. Extracts have displayed effectively reducing power and superoxide anion radical scavenging activity. Results of this study have highlighted potential antioxidant activity in the methanol and hexane extracts. The results of the phytochemical screening showed the presence of terpenoids and sterols with potential applications as food flavorants and functional foods, respectively. The use of Amphora sp. as a natural antioxidant source and a potential source of antibacterial compounds and phytochemicals in the food industry appears promising and should be investigated further.

Keywords: antioxidants, antimicrobial, microalgae, phytochemicals, cymbella

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Authors: Peramachi Sathiyamoorthy, Jacop Gopas, Avi Golan Goldhirsh

Abstract:

Corchorus olitorius is a leafy vegetable and an industrial crop. The herb has antioxidant, anti inflammatory, and anti-cancer properties. To assay the pharmaceutical properties, aqueous extracts of leaves and seeds from C. olitorius were tested against drug resistant melanoma cell line. The test showed LC50 of the extract was 0.08µg/ml. Aqueous seed extract exhibited higher melanoma inhibiting activity than leaf extract. Dialysis of seed extract showed that the active compound is less than 12 KDa. The compound with <3 KDa MW separated by microconcentration of seed extract showed 70.5 % inhibition of melanoma cell growth. Among the two fractions obtained by Gel filtration with G10 column, the first fraction at 1:2000 dilutions exhibited 100% inhibition of melanoma growth. The compound with Rf value 0.86 (MA4) isolated by TLC separation showed about 98% cytotoxicity against melanoma at 1: 1000 dilutions. Furthermore, HPLC separation of MA4 compound with Superdex 75 column resulted in 4 compounds. Out of 4, one compound showed melanoma inhibition. The active compound is identified by reagent methods as Strophanthidin. Further toxicological and clinical studies will lead to the development of a potential drug to treat drug resistant melanoma.

Keywords: corchorus olitorius, melanoma, drug development, strophanthidin

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Authors: A. Alao Olumuyiwa

Abstract:

Health hazards reported to be associated with exposure to electromagnetic radiations which include brain tumors, genotoxic effects, neurological effects, immune system deregulation, allergic responses and some cardiovascular effects are discussed under a closed tabular model in this study. This review however showed that there is strong and robust evidence that chronic exposures to electromagnetic frequency across the spectrum, through strength, consistency, biological plausibility and many dose-response relationships, may result in brain cancer and other carcinogenic disease symptoms. There is therefore no safe threshold because of the genotoxic nature of the mechanism that may however be involved. The discussed study explains that the cell phone has induced effects upon the blood –brain barrier permeability and the cerebellum exposure to continuous long hours RF radiation may result in significant increase in albumin extravasations. A physical Biomodeling approach is however employed to review this health effects using Specific Absorption Rate (SAR) of different GSM machines to critically examine the symptoms such as a decreased loco motor activity, increased grooming and reduced memory functions in a variety of animal spices in classified grouped and sub grouped models.

Keywords: brain cancer, electromagnetic radiations, physical biomodeling, specific absorption rate (SAR)

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Authors: M. Pourshirazi, M. Esmaelifar, A. Aliahmadi, F. Yazdian, A. S. Hatamian Zarami, S. J. Ashrafi

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Medicinal fungi are the new potential source of drugs to improve the treatment of diseases with association to oxidative agents such as cancers. Monascus purpureus contains functional components potentially effective in improving human health. In the present work, ethanolic extract of Monascus purpureus (EEM) was evaluated for health improving potential mainly focusing on antioxidant and anticancer activities. Ferric ion reducing power (FRAP), scavenging of DPPH radicals and determining viability of breast carcinoma MCF-7 and cervical carcinoma HeLa cells with MTT assay were evaluated. Our data showed a significant antioxidant activity of EEM with 142.45 µg/ml inhibition concentration of 50% DPPH radicals and 2112.33 µg eq.Fe2+/mg extract of FRAP assay. These results might be caused by antioxidant components such as pigments and phenolic compounds. Further, the results demonstrated that EEM caused significant reduction in the viability of MCF-7 with IC50 of 7 µg/ml but not have good effect against viability of HeLa cells. Accordingly, Monascus purpureus is presented as a strong potential of breast cancer treatment. In further study, the mechanistic studies are needed to determine the mechanisms of anticancer activity of EEM.

Keywords: Monascus purpureus, antioxidant, cancer, ethanolic extract

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Authors: Birgul Hizlar, Sibel Karakaya

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Nowadays, consumers are more and more aware of the benefits beyond basic nutrition provided by food and food compounds. Between these, carotenoids have been demonstrated to exhibit multiple health benefits (for example, some types of cancer, cardiovascular diseases, eye disorders, among others). However, carotenoid bioaccessibility and bioavailability is generally rather low due to their specific localization in plant tissue and lipophilic nature. This situation is worldwide issue, since both developed and developing countries have their interest and benefits in increasing the uptake of carotenoids from the human diet. Recently, a new class of foods designed to improve the bioaccessibility/bioavailability of orally administered bioactive compounds is introduced: excipient foods. Excipient foods are specially designed foods which are prepared depending on the physicochemical properties of target bioactive compounds and increasing the bioavailability or bioaccessibility of bioactive compound. In this study, effects of food matrix (greating, boiling and mashing) and different excipient foods (olive oil, lemon juice, whey curd and dried artichoke leaf powder) on bioaccessibility of β-carotene in carrot were investigated by means of simulating in vitro gastrointestinal (GI) digestion. β-carotene contents of grated, boiled and mashed (after boiling process) carrots were 79.28, 147.63 and 151.19 μg/g respectively. No significant differences among boiled and mashed samples indicated that mashing process had no effect on the release of β-carotene from the food matrix (p > 0.05). On the contrary, mashing causes significant increase in the β-carotene bioaccessibility (p < 0.05). The highest β-carotene content was found in the mashed carrots incorporated with olive oil and lemon juice (C2). However, no significant differences between that sample and C1 (mashed carrot with lemon juice, olive oil, dried artichoke leaf powder), C3 (mashed carrot with addition of olive oil, lemon juice, whey curd) and). Similarly, the highest β-carotene bioaccessibility (50.26%) was found mashed C3 sample (p < 0.05). The increase in the bioaccessibility was approximately 5 fold and 50 fold when compared to grated and mashed samples containing olive oil, lemon juice and whey curd. The results demonstrate that both, food matrix and excipient foods, are able to increase the bioaccessibility of β-carotene.

Keywords: bioaccessibility, carotenoids, carrot, β-carotene

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Authors: Panadda Rojpibulstit, Suthathip Kittisenachai, Songchan Puthong, Sirikul Manochantr, Pornpen Gamnarai, Sasichai Kangsadalampai, Sittiruk Roytrakul

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Hepatocellular carcinoma (HCC) is the most primary hepatic cancer worldwide. Nowadays a targeted therapy via monoclonal antibodies (mAbs) specific to tumor-associated antigen is continually developed in HCC treatment. In this regard, after establishing and consequently exploring Hep88 mAb’s tumoricidal effect on hepatocellular carcinoma cell line (HepG2 cell line), the Hep88 mAb’s specific Ag from both membrane and cytoplasmic fractions of HepG2 cell line was identified by 2-D gel electrophoresis and western blot analysis. After in-gel digestion and subsequent analysis by liquid chromatography-mass spectrometry (LC-MS), mortalin (HSPA9) and alpha-enolase were identified. The recombinant proteins specific to Hep88 mAb were cloned and expressed in E.coli BL21 (DE3). Moreover, alteration of HepG2 and Chang liver cell line after being induced by Hep88 mAb for 1-3 days was investigated using a transmission electron microscope. The result demonstrated that Hep88 mAb can bind to the recombinant mortalin (HSPA9) andalpha-enolase. In addition, gradual appearance of mitochondria vacuolization and endoplasmic reticulum dilatation were observed. Taken together, paraptosis-like programmed cell death (PCD) of HepG2 is induced by binding of mortalin (HSPA9) and alpha-enolase to Hep88 mAb. Mortalin depletion by formation of Hep88 mAb-mortalin (HSPA9) complex might initiate transcription-independent of p53-mediated apoptosis. Additionally, Hep88 mAb-alpha-enolase complex might initiate HepG2 cells energy exhaustion by glycolysis pathway obstruction. These results imply that Hep88 mAb might be a promising tool for development of an effective treatment of HCC in the next decade.

Keywords: Hepatocellular carcinoma, Monoclonal antibody, Paraptosis-like program cell death, Transmission electron microscopy, mortalin (HSPA9), alpha-enolase

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Authors: Esra Sengul, R. G. Aktas, M. E. Sitar, H. Isan

Abstract:

Hepatocellular carcinoma (HCC) is a hepatocellular tumor commonly found on the surface of the chronic liver. HepG2 is the most commonly used cell type in HCC studies. The main proteins remaining in the blood serum after separation of plasma fibrinogen are albumin and globulin. The fact that the albumin showed hepatocellular damage and reflect the synthesis capacity of the liver was the main reason for our use. Alpha-Fetoprotein (AFP) is an albumin-like structural embryonic globulin found in the embryonic cortex, cord blood, and fetal liver. It has been used as a marker in the follow-up of tumor growth in various malign tumors and in the efficacy of surgical-medical treatments, so it is a good protein to look at with albumins. We have seen the morphological changes of dimethyl sulfoxide (DMSO) on HepG2 and decided to investigate its biochemical effects. We examined the effects of DMSO, which is used in cell cultures, on albumin, AFP and total protein at low doses. Material Method: Cell Culture: Medium was prepared in cell culture using Dulbecco's Modified Eagle Media (DMEM), Fetal Bovine Serum Dulbecco's (FBS), Phosphate Buffered Saline and trypsin maintained at -20 ° C. Fixation of Cells: HepG2 cells, which have been appropriately developed at the end of the first week, were fixed with acetone. We stored our cells in PBS at + 4 ° C until the fixation was completed. Area Calculation: The areas of the cells are calculated in the ImageJ (IJ). Microscope examination: The examination was performed with a Zeiss Inverted Microscope. Daytime photographs were taken at 40x, 100x 200x and 400x. Biochemical Tests: Protein (Total): Serum sample was analyzed by a spectrophotometric method in autoanalyzer. Albumin: Serum sample was analyzed by a spectrophotometric method in autoanalyzer. Alpha-fetoprotein: Serum sample was analyzed by ECLIA method. Results: When liver cancer cells were cultured in medium with 1% DMSO for 4 weeks, a significant difference was observed when compared with the control group. As a result, we have seen that DMSO can be used as an important agent in the treatment of liver cancer. Cell areas were reduced in the DMSO group compared to the control group and the confluency ratio increased. The ability to form spheroids was also significantly higher in the DMSO group. Alpha-fetoprotein was lower than the values of an ordinary liver cancer patient and the total protein amount increased to the reference range of the normal individual. Because the albumin sample was below the specimen value, the numerical results could not be obtained on biochemical examinations. We interpret all these results as making DMSO a caretaking aid. Since each one was not enough alone we used 3 parameters and the results were positive when we refer to the values of a normal healthy individual in parallel. We hope to extend the study further by adding new parameters and genetic analyzes, by increasing the number of samples, and by using DMSO as an adjunct agent in the treatment of liver cancer.

Keywords: hepatocellular carcinoma, HepG2, dimethyl sulfoxide, cell culture, ELISA

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Authors: Vanja Misic, Mohamed El-Mogy, Yousef Haj-Ahmad

Abstract:

Endonuclease G (EndoG) is a well conserved mitochondrio-nuclear nuclease with dual lethal and vital roles in the cell. The aim of our study was to examine whether EndoG exerts its nuclease activity on exogenous DNA substrates such as plasmid DNA (pDNA), considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA and nuclease activity in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four day time-course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus, targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances it may non-specifically cleave intracellular DNA regardless of its origin. These findings make it unlikely that targeting of EndoG is a viable strategy for improving the duration and level of transgene expression from non-viral DNA vectors in gene therapy efforts.

Keywords: EndoG, silencing, exogenous DNA stability, HeLa cells

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Authors: Maria Zulema Morquecho Vega, Fabiola CarolinaMiranda Castro, Rafael Verdugo Miranda, Ignacio Yocupicio Villegas, Ana lidia Barron Raygoza, Martin enrique MArquez Cordova, Jose Alberto Duarte Moller

Abstract:

Background: Anemopsis californica, also called (tame grass) belongs to the Saururaceae family small, green plant. The blade is long and wide. Gives a white flower. The plant population is only found in humid, swampy habitats, it grows where there is water, along the banks of streams and water holes. In the winter, it dries up. The leaves, rhizomes, or roots of this plant have been used to treat a range of diseases. Some of its healing properties are used to treat wounds, cold and flu symptoms, spasmodic cough, infection, pain and inflammation, burns, swollen feet, as well as lung ailments, asthma, circulatory problems (varicose veins), rheumatoid arthritis, purifies blood, helps in urinary and digestive tract diseases, sores and healing, for headache, sore throat, diarrhea, kidney pain. The tea made from the leaves and roots is used to treat uterine cancer, womb cancer, relieves menstrual pain and stops excessive bleeding after childbirth. It is also used as a gynecological treatment for infections, hemorrhoids, candidiasis and vaginitis. Objective: To study the cytotoxicity of gels prepared with silver nanoparticles in AC extract combined with chitosan, collagen and hyaluronic acid as an alternative therapy for skin conditions. Methods: The Ag NPs were synthesized according to the following method. A 0.3 mg/mL solution is prepared in 10 ml of deionized water, adjust to pH 12 with NaOH, stirring is maintained constant magnetic and a temperature of 80 °C. Subsequently, 100 ul of a 0.1 M AgNO3 solution and kept stirring constantly for 15 min. Once the reaction is complete, measurements are performed by UV-Vis. A gel was prepared in a 5% solution of acetic acid with the respective nanoparticles and AC extract of silver in the extract of AC. Chitosan is added until the process begins to occur gel. At that time, collagen will be added in a ratio of 3 to 5 drops, and later, hyaluronic acid in 2% of the total compound formed. Finally, after resting for 24 hours, the cytotoxic effect of the gels was studied. in the presence of highly positive bacteria Staphylococcus aureus and highly negative for Escherichia coli. Cultures will be incubated for 24 hours in the presence of the compound and compared with the reference. Results: Silver nanoparticles obtained had a spherical shape and sizes among 20 and 30 nm. UV-Vis spectra confirm the presence of silver nanoparticles showing a surface plasmon around 420 nm. Finally, the test in presence of bacteria yield a good antibacterial property of this nanocompound and tests in people were successful. Conclusion: Gel prepared by biogenic synthesis shown beneficious effects in severe acne, acne vulgaris and wound healing with diabetic patients.

Keywords: anemopsis californica, nanomedicina, biotechnology, biomedicine

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