Search results for: cell survival

Authors: Amira, A. Goma, U. E. Mahrous

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Welfare may be considered to be a subjective experience; it has a biological function that is related to the fitness and survival of the animal accordingly, researches have suggested that welfare is compromised when the animal's evolutionary fitness is reduced. This study was carried out to explain the effect of some managerial stressors as handling and restraint on behavioral patterns and biochemical parameters of rats. A total of 24 (12 males and 12 females) Sprague-Dawley rats (12 months and 150-180g) were allotted into 3 groups, handled group (4 male and 4 female), restrained group (4 male and 4 female) and control group (4 males and 4 females). The obtained results revealed that time spent feeding, drinking frequency, movement and cage exploration increased significantly in handled rats than other groups, while lying time and licking increased significantly in restrained rats than handled and controls. Moreover, social behavior decreased in both stressed groups than control. Triglycerides were significantly increased in handled rats than other groups, while total lipid, total protein and globulin significantly increased in both treated groups than control. Corticosterone increased in restrained and handled rats than control ones. Moreover, there was an increment in packed cell volume significantly in restrained rats than others. These deducted that if we want to study the effect of stress on animal welfare it is necessary to study the effect of such stressors on animal’s behavior and physiological responses.

Keywords: handling, restraint, welfare, rat, behavior, physiology

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Authors: Yemane Hailu Fissuh, Zhongzhan Zhang

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In biomedical researches and randomized clinical trials, the most commonly interested outcomes are time-to-event so-called survival data. The importance of robust models in this context is to compare the effect of randomly controlled experimental groups that have a sense of causality. Causal estimation is the scientific concept of comparing the pragmatic effect of treatments conditional to the given covariates rather than assessing the simple association of response and predictors. Hence, the causal effect based semiparametric transformation model was proposed to estimate the effect of treatment with the presence of possibly time-varying covariates. Due to its high flexibility and robustness, the semiparametric transformation model which shall be applied in this paper has been given much more attention for estimation of a causal effect in modeling left-truncated and right censored survival data. Despite its wide applications and popularity in estimating unknown parameters, the maximum likelihood estimation technique is quite complex and burdensome in estimating unknown parameters and unspecified transformation function in the presence of possibly time-varying covariates. Thus, to ease the complexity we proposed the modified estimating equations. After intuitive estimation procedures, the consistency and asymptotic properties of the estimators were derived and the characteristics of the estimators in the finite sample performance of the proposed model were illustrated via simulation studies and Stanford heart transplant real data example. To sum up the study, the bias of covariates was adjusted via estimating the density function for truncation variable which was also incorporated in the model as a covariate in order to relax the independence assumption of failure time and truncation time. Moreover, the expectation-maximization (EM) algorithm was described for the estimation of iterative unknown parameters and unspecified transformation function. In addition, the causal effect was derived by the ratio of the cumulative hazard function of active and passive experiments after adjusting for bias raised in the model due to the truncation variable.

Keywords: causal estimation, EM algorithm, semiparametric transformation models, time-to-event outcomes, time-varying covariate

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Authors: Mohd Shazad, Kamal Kumar Gupta

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Aedes aegypti (Diptera: Culicidae), commonly known as tiger mosquito is the vector of dengue fever, yellow fever, chikungunya and zika virus. In the absence of any effective vaccine against these diseases, control the mosquito population is the only promising mean to prevent the diseases. Currently used chemical insecticides cause environmental contamination, high mammalian toxicity and hazards to non-target organisms, insecticide resistance and vector resurgence. Present research work aimed to explore the potentials of phytochemicals present in the Ocimum sanctum in management of mosquito population. The leaves of Ocimum were extracted with ethanol by ‘cold extraction method’. 0-24h old fourth instar larvae of Aedes aegypti were treated with the extract of concentrations 50ppm, 100ppm, 200ppm and 400ppm for 24h. Survival, growth and development of the treated larvae were evaluated. The adults emerged from the treated larvae were used for the reproductive fitness studies. Our results indicate 77.2% mortality in the larvae exposed to 400 ppm. At lower doses, although there was no significant reduction in the survival after 24h however, it decreased during subsequent days of observations. In control experiments, no mortality was observed. It was also observed that the larvae survived after treatment showed severe growth and developmental abnormalities. There was significant increase in larval duration. In control, fourth instar moulted into pupa after 3 days while larvae treated with 400 ppm extract were moulted after 4.6 days. Larva-pupa intermediates and the pupa-adult intermediates were observed in many cases. The adults emerged from the treated larvae showed impaired mating and oviposition behaviour. The females exhibited longer preoviposition period, reduced oviposition rate and decreased egg output. GCMS analysis of the ethanol extract revealed presence of JH mimics and intermediates of JH biosynthetic pathway. Potentials of Ocimum sanctum in integrated vector management programme of Aedes aegypti were discussed.

Keywords: Aedes aegypti, Ocimum sanctum, oviposition, survival

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Authors: James V. Luisi

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Understanding consciousness requires a synthesis of ideas from multiple disciplines, including obvious ones like psychology, biology, evolution, neurology, and neuroscience, as well as less obvious ones like protozoology, botany, entomology, carcinology, herpetology, mammalogy, and computer sciences. Furthermore, to incorporate the necessary backdrop, it is best presented in a theme of Eastern philosophy, specifically leveraging the teachings of Buddhism for its relevance to early thought on consciousness. These ideas are presented as a multi-level framework that illustrates the various aspects of consciousness within a tapestry of foundational and dependent building blocks as to how living organisms evolved to understand elements of their reality sufficiently to survive, and in the case of Homo sapiens, eventually move beyond meeting the basic needs of survival, but to also achieve survival of the species beyond the eventual fate of our planet. This is not a complete system of thought, but just a framework of consciousness gathering some of the key elements regarding the evolution of consciousness and the advent of free will, and presenting them in a unique way that encourages readers to continue the dialog and thought process as an experience to enjoy long after reading the last page. Readers are encouraged to think for themselves about the issues raised herein and to question every facet presented, as much further exploration is needed. Needless to say, this subject will remain a rapidly evolving one for quite some time to come, and it is probably in the interests of everyone to at least consider attaining both an ability and willingness to participate in the dialog.

Keywords: consciousness, sentience, intelligence, artificial intelligence, Buddhism

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Authors: Aliakbar Hafezi, Jalal Taherian, Jamshid Abedi, Mahsa Elahi

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Background: Bones are commonly affected by renal cell carcinoma (RCC) (primarily or secondary), and this condition causes bone fragility. The aim of this study was to evaluate the diagnostic value of noninvasive methods for the diagnosis of ROD in RCC patients on hemodialysis (HD) in northern Iran. Methods: In this cross-sectional study, 50 RCC patients with ESRD referred to dialysis units in northern Iran during 2021-2024 were randomly selected and investigated. The biochemical and radiographic evaluation of ROD and its subtypes was performed, and then all patients underwent bone biopsy and histopathological study, and finally, the diagnostic value of the noninvasive methods was assessed. Results: The mean age of patients was 58.9 ± 11.7 years, and 27 cases (54.0%) were female. 38 (76.0%) of RCC patients with ESRD had ROD, and 12 patients (24.0%) had no evidence of bone disorders. The sensitivity, specificity, positive and predictive values and accuracy of the noninvasive methods for the diagnosis of ROD were 92%, 82%, 95%, 75% and 90%, respectively. Conclusion: This study showed that the frequency of ROD in RCC patients with ESRD in northern Iran was high and the biochemical and radiographic markers have a high diagnostic value for ROD as well as histopathological assessment.

Keywords: renal cell carcinoma, renal osteodystrophy, hemodialysis, non-metastatic

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Authors: Marcel Verwey, Anna M. Joubert, Elsie M. Nolte, Wolfgang Dohle, Barry V. L. Potter, Anne E. Theron

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A tetrahydroisoquinolinone (THIQ) core can be used to mimic the A,B-ring of colchicine site-binding microtubule disruptors such as 2-methoxyestradiol in the design of anti-cancer agents. Steroidomimeric microtubule disruptors were synthesized by introducing C'2 and C'3 of the steroidal A-ring to C'6 and C'7 of the THIQ core and by introducing a decorated hydrogen bond acceptor motif projecting from the steroidal D-ring to N'2. For this in vitro study, four non-steroidal THIQ-based analogues were investigated and comparative studies were done between the non-sulphamoylated compound STX 3450 and the sulphamoylated compounds STX 2895, STX 3329 and STX 3451. The objective of this study was to investigate the modes of cell death induced by these four THIQ-based analogues in A549 lung carcinoma epithelial cells and metastatic breast adenocarcinoma MDA-MB-231 cells. Cytotoxicity studies to determine the half maximal growth inhibitory concentrations were done using spectrophotometric quantification via crystal violet staining and lactate dehydrogenase (LDH) assays. Microtubule integrity and morphologic changes of exposed cells were investigated using polarization-optical transmitted light differential interference contrast microscopy, transmission electron microscopy and confocal microscopy. Flow cytometric quantification was used to determine apoptosis induction and the effect that THIQ-based analogues have on cell cycle progression. Signal transduction pathways were elucidated by quantification of the mitochondrial membrane integrity, cytochrome c release and caspase 3, -6 and -8 activation. Induction of autophagic cell death by the THIQ-based analogues was investigated by morphological assessment of fluorescent monodansylcadaverine (MDC) staining of acidic vacuoles and by quantifying aggresome formation via flow cytometry. Results revealed that these non-steroidal microtubule disrupting analogues inhibited 50% of cell growth at nanomolar concentrations. Immunofluorescence microscopy indicated microtubule depolarization and the resultant mitotic arrest was further confirmed through cell cycle analysis. Apoptosis induction via the intrinsic pathway was observed due to depolarization of the mitochondrial membrane, induction of cytochrome c release as well as, caspase 3 activation. Potential involvement of programmed cell death type II was observed due to the presence of acidic vacuoles and aggresome formation. Necrotic cell death did not contribute significantly, indicated by stable LDH levels. This in vitro study revealed the induction of the intrinsic apoptotic pathway as well as possible involvement of autophagy after exposure to these THIQ-based analogues in both MDA-MB-231- and A549 cells. Further investigation of this series of anticancer drugs still needs to be conducted to elucidate the temporal, mechanistic and functional crosstalk mechanisms between the two observed programmed cell deaths pathways.

Keywords: apoptosis, autophagy, cancer, microtubule disruptor

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Authors: Alieh Farshbaf, Tayyeb Bahrami

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Introduction: Cc-chemokine receptor-5 (CCR5) is known as a main co-receptor in human immunodeficiency virus type-1 (HIV-1) infection. Many studies showed 32bp deletion (Δ32) in CCR5 gene, provide natural resistance to HIV-1 infection in homozygous individuals. Inducing the resistance mechanism by CCR5 in HIV-1 infected patients eliminated many problems of highly-active-anti retroviral therapy (HAART) drugs like as low safety, side-effects and virus rebounding from latent reservoirs. New treatments solved some restrictions that are based on gene modification and cell therapy. Literature review: The stories of the “Berlin and Boston patients” showed autologous hematopoietic stem cells transplantation (HSCT) could provide effective cure of HIV-1 infected patients. Furthermore, gene modification by zinc finger nuclease (ZFN) demonstrated another successful result again. Despite the other studies for gene therapy by ∆32 genotype, there is another mutation -CCR5 ∆32/m303- that provides HIV-1 resistant. It is a heterozygote genotype for ∆32 and T→A point mutation at nucleotide 303. These results approved the key role of CCR5 gene. Conclusion: Recent studies showed immune gene therapy and cell therapy could provide effective cure for refractory disease like as HIV. Eradication of HIV-1 from immune system was not observed by HAART, because of reloading virus genome from latent reservoirs after stopping them. It is showed that CCR5 could induce natural resistant to HIV-1 infection by the new approaches based on stem cell transplantation and gene modifying.

Keywords: CCR5, HIV-1, stem cell, immune gene therapy, gene modification

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Authors: Michela Terlizzi, Chiara Colarusso, Aldo Pinto, Rosalinda Sorrentino

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Sphingosine-1-phosphate (S1P) is a membrane-derived bioactive phospholipid exerting a multitude of effects on respiratory cell physiology and pathology through five S1P receptors (S1PR1-5). Higher levels of S1P have been registered in a broad range of respiratory diseases, including inflammatory disorders and cancer, although its exact role is still elusive. Based on our previous study in which we found that S1P/S1PR3 is involved in an inflammatory pattern via the activation of Toll-like Receptor 9 (TLR9), highly expressed on lung cancer cells, the main goal of the current study was to better understand the involvement of S1P/S1PR3 pathway/signaling during lung carcinogenesis, taking advantage of a mouse model of first-hand smoke exposure and of carcinogen-induced lung cancer. We used human samples of Non-Small Cell Lung Cancer (NSCLC), a mouse model of first-hand smoking, and of Benzo(a)pyrene (BaP)-induced tumor-bearing mice and A549 lung adenocarcinoma cells. We found that the intranuclear, but not the membrane, localization of S1PR3 was associated to the proliferation of lung adenocarcinoma cells, the mechanism that was correlated to human and mouse samples of smoke-exposure and carcinogen-induced lung cancer, which were characterized by higher utilization of S1P. Indeed, the inhibition of the membrane S1PR3 did not alter tumor cell proliferation after TLR9 activation. Instead, according to the nuclear localization of sphingosine kinase (SPHK) II, the enzyme responsible for the catalysis of the S1P last step synthesis, the inhibition of the kinase completely blocked the endogenous S1P-induced tumor cell proliferation. These results prove that the endogenous TLR9-induced S1P can on one side favor pro-inflammatory mechanisms in the tumor microenvironment via the activation of cell surface receptors, but on the other tumor progression via the nuclear S1PR3/SPHK II axis, highlighting a novel molecular mechanism that identifies S1P as one of the crucial mediators for lung carcinogenesis-associated inflammatory processes and that could provide differential therapeutic approaches especially in non-responsive lung cancer patients.

Keywords: sphingosine-1-phosphate (S1P), S1P Receptor 3 (S1PR3), smoking-mice, lung inflammation, lung cancer

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Authors: Noraziah Nordin, Najihah Mohd Hashim, Nazia Abdul Majid, Mashitoh Abdul Rahman, Hamed Karimian, Hapipah Mohd Ali

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Enicosanthellum pulchrum belongs to family Annonaceae is also known as family of 'mempisang' in Malaysia. Liriodenine was isolated by prep-HPLC method. This method was first technique used for the isolation of this compound. The structure of the liriodenine was elucidated by 1D and 2D spectroscopy techniques. Liriodenine was tested on ovarian cancer cells line (CAOV-3) for MTT, AO/PI and cytotoxicity 3 assays. The MTT assay was performed to determine the cytotoxicity effect of lirodenine on CAOV-3 cells. The morphological changes on CAOV-3 cells were observed by AO/PI assay for the early and late stage of apoptosis, as well as necrosis. Meanwhile, the measurement of cell loss, nuclear morphology, DNA content, cell membrane permeability, mitochondrial membrane potential changes and cytochrome c release from mitochondria were detected through cytotoxicity 3 assay. The IC50 results showed liriodenine inhibits the growth of CAOV-3 cells after 24 h of treatment at 10.25 ± 1.06 µg/mL. After 48 and 72 h of treatments, the IC50 values were decreased to 7.65 ± 0:07 and 6.35 ± 1.62 µg/mL, respectively. The morphology changes can be seen on CAOV-3 with a production of cell membrane blebbing, cromatin condensation and apoptotic bodies with increasing time of treatment from 24 to 72 h. Evaluation of cytotoxicity 3 on CAOV-3 cells after treated with liriodenine, resulting loss of mitochondrial membrane potential and release of cytochrome c from mitochondria. The results demonstrated the capability of liriodenine as a promising anticancer agent, particularly on human ovarian cancer.

Keywords: Enicosanthellum pulchrum, ovarian cancer, apoptosis, cytotoxicity

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Authors: H. Chassaigne, S. Gioria, J. Lobo Vicente, D. Carpi, P. Barboro, G. Tomasi, A. Kinsner-Ovaskainen, F. Rossi

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Emerging approaches in the area of exposure to nanomaterials and assessment of human health effects combine the use of in vitro systems and analytical techniques to study the perturbation of the proteome and/or the metabolome. We investigated the modification in the cytoplasmic compartment of the Balb/3T3 cell line exposed to gold nanoparticles. On one hand, the proteomic approach is quite standardized even if it requires precautions when dealing with in vitro systems. On the other hand, metabolomic analysis is challenging due to the chemical diversity of cellular metabolites that complicate data elaboration and interpretation. Differentially expressed proteins were found to cover a range of functions including stress response, cell metabolism, cell growth and cytoskeleton organization. In addition, de-regulated metabolites were annotated using the HMDB database. The "omics" fields hold huge promises in the interaction of nanoparticles with biological systems. The combination of proteomics and metabolomics data is possible however challenging.

Keywords: data processing, gold nanoparticles, in vitro systems, metabolomics, proteomics

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Authors: Diwei Lin, Amanda Jia Hui Tan

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In 2012, testicular cancer was estimated to account for 940 disability adjusted life years in Australia; of these, 450 were years lost due to premature death and 500 were years of healthy life lost due to disease, disability or injury. Testicular choriocarcinoma is one of the rarest variants of testicular germ cell tumours, accounting for less than 1% of testicular germ cell tumours and only about 0.19% of all testicular tumours. Management involves radical orchiectomy followed by chemotherapy. Even then, the prognosis is extremely poor. This case report describes a 20-year-old male with pure testicular choriocarcinoma with pulmonary metastases.

Keywords: testicular cancer, choriocarcinoma, cryptorchidism, chemotherapy, metastatic testicular cancer

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Authors: Yusser Olguín, Diego Benavente, Fernando Dorta, Nicole Orellana, Cristian Acevedo

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One of the greatest challenges of tissue engineering is the generation of materials in which the highest possible number of conditions can be incorporated to stimulate the proliferation and differentiation of cells, which will be transformed together with the material into new functional tissue. In this sense, considering the properties of microfluidics and its relationship with cellular micro-environments, the possibility of controlling flow patterns and the ability to design diverse patterns in the chips, a microfluidic cell culture system can be established as a means for the evaluation of the effect of different parameters in a controlled and precise manner. Specifically in relation to the study and development of alternatives in peripheral nervous tissue engineering, it is necessary to consider different physical and chemical neurotrophic stimuli that promote cell growth and differentiation. Chemical stimuli include certain vitamins, glucocorticoids, gangliosides, and growth factors, while physical stimuli include topological stimuli, mechanical forces of the cellular environment and electrical stimulation. In this context, the present investigation shows the results of cell stimulation in a microbioreactor using electrical and chemical stimuli, where the differentiation of PC12 cells as a neuronal model is evidenced by neurite expression, dependent on the stimuli and their combination. The results were analysed with a multi-factor statistical approach, showing several relationships and dependencies between different parameters. Chip design, operating parameters and concentrations of neurotrophic chemical factors were found to be preponderant, based on the characteristics of the electrical stimuli.

Keywords: microfluidics, nerve tissue engineering, microbioreactor, electrical stimuli

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Authors: Mi-Hye Hwang, Young Min Son, Kichan Lee, Bang-Hun Hyun, Byeong Yeal Jung

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Botulism is a paralytic disease of human beings and animals caused by neurotoxin produced by Clostridium botulinum. The neurotoxins are genetically distinguished into 8 types, A to H. Ingestion of performed toxin, usually types B, C, and D, have been shown to produce diseases in most cases of cattle botulism. Vaccination is the best measure to prevent cattle botulism. However, the commercially available toxoid-based vaccines are difficult and hazardous to produce. We produced recombinant protein using gene of heavy chain domain of botulinum toxin B of which binds to cellular receptor of neuron cells and used as immunogen. In this study, we evaluated the safety and efficacy of botulism vaccine composed of recombinant types B. Safety test was done by National Regulation for Veterinary Biologicals. For efficacy test, female ICR mice (5 weeks old) were subcutaneously injected, intraperitoneally challenged, and examined the survival rates compared with vaccination and non-vaccination group. Mouse survival rate of recombinant types B vaccine was above 80%, while one of non-vaccination group was 0%. A vaccine composed of recombinant types B was safe and efficacious in mouse. Our results suggest that recombinant heavy chain receptor binding domain can be used as an effective vaccine candidate for type B botulism.

Keywords: botulism, livestock, vaccine, recombinant protein, toxin

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Authors: Ya-Ting Chuang, Krystle Leung, Ya-Jen Chang, Rosemarie H. DeKruyff, Paul B. Savage, Richard Cruse, Christophe Benoit, Dirk Elewaut, Nicole Baumgarth, Dale T. Umetsu

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We examined characteristics of a Natural Killer T (NKT) cell subpopulation that developed during influenza infection in neonatal mice, and that suppressed the subsequent development of allergic asthma in a mouse model. This NKT cell subset expressed CD38 but not CD4, produced IFN-γ, but not IL-17, IL-4 or IL-13, and inhibited the development of airway hyperreactivity (AHR) through contact-dependent suppressive activity against helper CD4 T cells. The NKT subset expanded in the lungs of neonatal mice after infection with influenza, but also after treatment of neonatal mice with a Th1-biasing α-GalCer glycolipid analogue, Nu-α-GalCer. These results suggest that early/neonatal exposure to infection or to antigenic challenge can affect subsequent lung immunity by altering the profile of cells residing in the lung and that some subsets of NKT cells can have direct inhibitory activity against CD4+ T cells in allergic asthma. Importantly, our results also suggest a potential therapy for young children that might provide protection against the development of asthma.

Keywords: NKT subset, asthma, airway hyperreactivity, hygiene hypothesis, influenza

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Authors: Lemuel David

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This study aims to investigate The Impact of Tax Policies on Small Business Growth in Developing Countries: A Case Study of Montserrado Mount County, Republic of Liberia. Businesses in Liberia are crucial for job creation and the economic empowerment of its citizens, especially in Grand Cape Mount County where they provide 95% of all jobs and support local capital formation. However, many businesses face challenges that lead to premature closure, including tax-related issues such as multiple taxations and high tax burdens. This research aims to examine the effects of various taxation on business survival in Grand Cape Mount County. The study employed a survey research design with a population of 50 and a sample size of 74. Data was collected using a self-administered questionnaire and analyzed quantitatively with simple percentages, and the research hypotheses were tested with ANOVA. The study findings revealed that multiple taxations hurts business survival, and the relationship between business size and its ability to pay taxes is significant. Therefore, the study recommends that the government of Liberia should create uniform tax policies that support business development in Grand Cape Mount County, and consider the size of businesses when formulating tax policies.

Keywords: multiple taxations, businesses, mortality, growth

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Authors: Ila Shukla, Lubna Azmi, Shyam Sunder Gupta, C. V. Rao

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Treatments against Hepatitis-C virus (HCV) are already available, but the current high cost of such treatments limit them to wealthy patients only. Hence our current study is aimed at the rectification of HCV infection by using Lichen rangiferinus (LRE) extract in in vitro cultures. Anti-HCV activity of the given extract was evaluated using the virus grown in cell culture (HCVcc). Two control inhibitors, erlotinib and telaprevir, were systematically included in each experiment. At the end of the incubation period, we evaluated cell viability and viral replication. The LRE inhibited the growth of HCV in a dose dependent manner.

Keywords: Erlotinib, Hepatitis C, Lichen rangiferinus, Telaprevir

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Authors: Kheireddine El-Boubbou

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Drug delivery to target human acute myeloid leukemia (AML) using a nanoparticulate chemotherapeutic formulation that can deliver drugs selectively to AML cancer is hugely needed. In this work, we report the development of a nanoformulation made of polymeric-stabilized multifunctional magnetic iron oxide nanoparticles (PMNP) loaded with the anticancer drug Doxorubicin (Dox) as a promising drug carrier to treat AML. Dox@PMNP conjugates simultaneously exhibited high drug content, maximized fluorescence, and excellent release properties. Nanoparticulate uptake and cell death following addition of Dox@PMNPs were then evaluated in different types of human AML target cells, as well as on normal human cells. While the unloaded MNPs were not toxic to any of the cells, Dox@PMNPs were found to be highly toxic to the different AML cell lines, albeit at different inhibitory concentrations (IC₅₀ values), but showed very little toxicity towards the normal cells. In comparison, free Dox showed significant potency concurrently to all the cell lines, suggesting huge potentials for the use of Dox@PMNPs as selective AML anticancer cargos. Live confocal imaging, fluorescence and electron microscopy confirmed that Dox is indeed delivered to the nucleus in relatively short periods of time, causing apoptotic cell death. Importantly, this targeted payload may potentially enhance the effectiveness of the drug in AML patients and may further allow physicians to image leukemic cells exposed to Dox@PMNPs using MRI.

Keywords: magnetic nanoparticles, drug delivery, acute myeloid leukemia, iron oxide, cancer nanotherapy

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Authors: Dileep Kumar Verma, Sunil Kumar Lal

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Influenza still remains one of the most challenging diseases posing significant threat to public health causing seasonal epidemics and pandemics. Influenza A Virus (IAV) surface protein hemagglutinin is known to play an important role in viral attachment to the host sialic acid receptors and concentrate in lipid rafts for efficient viral fusion. Selective nature of Influenza A virus to utilize rafts micro-domain for efficient virus assembly and budding has been explored in depth. However, the detailed mechanism of IAV binding to host cell membrane and entry into the host remains elusive. In the present study we investigated the role of lipid rafts in early life cycle events of IAV. Role of host lipid rafts was studied using raft disruption method by extraction of cholesterol by Methyl-β-Cyclodextrin. Using GM1, a well-known lipid raft marker, we were able to observe co-localization of IAV on lipid rafts on the host cell membrane. This experiment suggests a direct involvement of lipid rafts in the initiation of the IAV life cycle. Upon disruption of lipid rafts by Methyl-b-cyclodextrin, we observed a significant reduction in IAV binding on the host cell surface indicating a significant decrease in virus attachment to coherent membrane rafts. Our results provide proof that host lipid rafts and their constituents play an important role in the adsorption of IAV. This study opens a new avenues in IAV virus-host interactions to combat infection at a very early steps of the viral lifecycle.

Keywords: lipid raft, adsorption, cholesterol, methyl-β-cyclodextrin, GM1

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Authors: Benchouala Amira, Bojic Clement, Poupin Pascal, Cossu Leguille-carole

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The objective of this study is to evaluate in vitro the cytotoxic and androgenic potential of several antifungal molecules (amphotericin B, econazole, ketoconazole and miconazole) on MDA-Kb2 cell lines. This biological model is an effective tool for the detection of endocrine disruptors because it responds well to the main agonist of the androgen receptor (testosterone) and also to an antagonist: flutamide. The cytotoxicity of each chemical compound tested was measured using an MTT assay (tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) which measures the activity of the reductase function of mitochondrial succinate dehydrogenase enzymes of cultured cells. This complementary cytotoxicity test is essential to ensure that the effects of reduction in luminescence intensity observed during androgenic tests are only attributable to the anti-androgenic action of the compounds tested and not to their possible cytotoxic properties. Tests of the androgenic activity of antifungals show that these compounds do not have the capacity to induce transcription of the luciferase gene. These compounds do not exert an androgenic effect on MDA-Kb2 cells in culture for the environmental concentrations tested. The addition of flutamide for the same tested concentrations of antifungal molecules reduces the luminescence induced by amphotericin B, econazole and miconazole, which is explained by a strong interaction of these molecules with flutamide which may have a greater toxic effect than when tested alone. The cytotoxicity test shows that econazole and ketoconazole can cause cell death at certain concentrations tested. This cell mortality is perhaps induced by a direct or indirect action on deoxyribonucleic acid (DNA), ribonucleic acid (RNA) or proteins necessary for cell division.

Keywords: cytotoxicity, androgenic potential, antifungals, MDA-Kb2

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Authors: Athanasios A. Tountas, Geoffrey A. Ozin, Mohini M. Sain

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Renewable hydrogen (H₂) carriers such as methanol (MeOH), dimethyl ether (DME), oxymethylene dimethyl ethers (OMEs), and conceivably ammonia (NH₃) can be reformed back into H₂ and are fundamental chemical conversions for the long-term viability of the H₂ economy due to their higher densities and ease of transportability compared to H₂. MeOH is an especially important carrier as it is a simple C1 chemical that can be produced from green solar-PV-generated H₂ and direct-air-captured CO₂ with a current commercially practical solar-to-fuel efficiency of 10% from renewable solar energy. MeOH steam reforming (MSR) in stationary systems next to H₂ fuel-cell filling stations can eliminate the need for onboard mobile reformers, and the former systems can be more robust in terms of attaining strict H₂ product specifications, and MeOH is a safe, lossless, and compact medium for long-term H₂ storage. Both thermal- and photo-catalysts are viable options for achieving the stable, long-term performance of stationary MSR systems.

Keywords: fuel-cell vehicle filling stations, methanol steam reforming, hydrogen transport and storage, stationary reformer, liquid hydrogen carriers

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Authors: Cristian Soitu, Alexander Feuerborn, Cyril Deroy, Alfonso Castrejon-Pita, Peter R. Cook, Edmond J. Walsh

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Microfluidics now stands as an academically mature technology after a quarter of a century research activities have delivered a vast array of proof of concepts for many biological workflows. However, translation to industry remains poor, with only a handful of notable exceptions – e.g. digital PCR, DNA sequencing – mainly because of biocompatibility issues, limited range of readouts supported or complex operation required. This technology exploits the domination of interfacial forces over gravitational ones at the microscale, replacing solid walls with fluid ones as building blocks for cell micro-environments. By employing only materials used by biologists for decades, the system is shown to be biocompatible, and easy to manufacture and operate. The method consists in displacing a continuous fluid layer into a pattern of isolated chambers overlaid with an immiscible liquid to prevent evaporation. The resulting fluid arrangements can be arrays of micro-chambers with rectangular footprint, which use the maximum surface area available, or structures with irregular patterns. Pliant, self-healing fluid walls confine volumes as small as 1 nl. Such fluidic structures can be reconfigured during the assays, giving the platform an unprecedented level of flexibility. Common workflows in cell biology are demonstrated – e.g. cell growth and retrieval, cloning, cryopreservation, fixation and immunolabeling, CRISPR-Cas9 gene editing, and proof-of-concept drug tests. This fluid-shaping technology is shown to have potential for high-throughput cell- and organism-based assays. The ability to make and reconfigure on-demand microfluidic circuits on standard Petri dishes should find many applications in biology, and yield more relevant phenotypic and genotypic responses when compared to standard microfluidic assays.

Keywords: fluid walls, micro-chambers, reconfigurable, freestyle

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Authors: Saekil Yun, Sib Sankar Giri, Jin Woo Jun, Hyoun Joong Kim, Sang Guen Kim, Sang Wha Kim, Jung Woo Kang, Se Jin Han, Se Chang Park

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In aquaculture, vaccination is important to control and prevent diseases. In the study, we utilized poly D,L-lactide-co-glycolic acid (PLGA) microparticles (MPs) for encapsulating formalin-killed Aeromonas hydrophila cells. To assess the innate and adaptive immune responses, carps and loaches were used for the experiments. Fish were divided into three groups (A, B, C). Total antigen of 0.1 ml vaccine was adjusted by 2 x 108 CFU and injected via intraperitoneal route. Group A was vaccinated with 0.1 ml of PLGA vaccine, group B was with 0.1 ml of FKC vaccine and group C was with 0.1 ml of sterile PBS. All three groups were challenged with A. hydrophila and challenge dose was lethal dose (LD50). Loaches and carp were then challenged with A. hydrophila at 12 and 20 weeks post vaccination (wpv), and 10 and 14 wpv, respectively, and relative survival rates were calculated. For both fish species, the curve of antibody titer over time was shallower in the PLGA group than the FKC group and the PLGA groups demonstrated higher survival rates at all time-points. In the groups of PLGA-MP, relative mRNA levels of IL-1β, TNF-α, lysozyme C and IgM were significantly upregulated than FKC treated groups. Biodegradable PLGA microparticle vaccine could induce longer immune responses than original FKC vaccines to protect from A. hydrophila infection.

Keywords: PLGA, microparticles, Aeromonas hydrophila, vaccine

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Authors: Laila Seada, Nouf Al Gharbi, Shaimaa Dawa

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Although skin cancers are prevalent worldwide, it is uncommon in Ha’il region in the Kingdom of Saudi Arabia, mostly non-melanoma sub-type. During a 4-year period from 2014 to 2017, out of a total of 120 cases of skin lesions, 29 non-melanoma cancers were retrieved from histopathology files obtained from King Khalid Hospital. As part of the study, all cases of skin cancer diagnosed during 2014 -2017 have been revised and the clinicopathological data recorded. The results show that Basal cell carcinoma (BCC) was the most common neoplasm (36%), followed by cutaneous lymphomas (mostly mycosis fungoides 25%), squamous cell carcinoma (SCC) (21%) and dermatofibrosarcoma protuberans (DFSP) (11%). Only one case of metastatic carcinoma was recorded. BCC nodular type was the most prevalent, with a mean age 57.6 years and mean size 2.73 cm. SCC was mostly grade 2, with mean size 1.9 cm and an older mean age of 72.3 cm. Increased size of lesion positively correlated with older age (p = 0.001). Non-melanoma skin cancer in Ha’il region is not frequently encountered. BCC is the most frequent followed by cutaneous T-cell lymphomas and SCC. The findings in this study were in accordance with other parts of, but much lower than other parts of the world.

Keywords: non melanoma skin cancer, Hail Region, histopathology, BCC

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Authors: Sajjad Ali Khan Sajjad Ali Khan, Gohar Ayub, Khalil Ur Rahman, Muhammad Sajid, Mumtaz Farooq, Mohammad Irshad, Haider Ali

Abstract:

A trail was carried out to know the effect of different soil media and planting time on the cuttings of cherry (Prunus avium L.) rootstock Colt at Agriculture Research Institute (ARI) Mingora swat, during winter 2011. The experiment was laid out in Randomized Complete Block Design (RCBD) with split plot arrangement and was replicated three times. Soil media (Silt, Garden soil and Silt+Garden soil+FYM) were assigned to main plots whereas, planting Dates (1st Jan, 11th Jan, 21st Jan, 1st Feb, 11th Feb, 21st Feb and 2nd March) subjected to sub plots. The data recorded on sprouting percentage, shoot diameter cutting-1, number of leaves cutting-1, rootstock height (cm), survival percentage, number of roots, root length (cm), root volume (cm3) and root weight (gm) were significantly affected by different soil media. Maximum sprouting percentage (100%), shoot diameter (1.72 mm), number of leaves cutting-1 (76.74), rootstock height (104.36 cm), survival percentage (41.67%), number of roots (76.35), root length (11.28 cm), root volume (4.43 cm3) and root weight (4.64 gm) were recorded in media M3 (Garden soil+silt+FYM). A significant response to various planting dates were observed for most of vegetative and rooting attributes of cherry rootstock Colt. 1st January plantation showed maximum sprouting percentage (100%), shoot diameter (1.99 mm), number of leaves (81.46), rootstock height (126.24 cm), survival percentage (58.12%), whereas 11th January plantation showed more number of roots (94.43), root length (10.60 cm), root volume (3.68 cm3) and root weight (3.71 gm). Based on the results from the experimental work, it is recommended that cherry cuttings should be planted in early January in soil media (Silt+Garden soil+ FYM) for better growth and development under the agro climatic conditions of temperate region.

Keywords: soil media, cherry rootstock, planting dates, growth parameters

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Authors: Reshu Saxena, R. K. Tripathi

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HIV-1 transmission and spread involves significant host-virus interaction. Potential targets for prevention of HIV-1 lies at the site of mucosal barriers. Thus a better understanding of how HIV-1 infects target cells at such sites and lead their invasion is required, with prime focus on the host determinants regulating HIV-1 spread. HIV-1 Nef is important for viral infectivity and pathogenicity. It promotes HIV-1 replication, facilitating immune evasion by interacting with various host factors and altering cellular pathways via multiple protein-protein interactions. In this study nef was sequenced from HIV-1 patients, and showed specific mutations revealing sequence variability in nef. To explore the difference in Nef functionality based on sequence variability we have studied the effects of HIV-1 Nef in human SupT1 T cell line and (THP-1) monocyte-macrophage cell lines through proteomics approach. 2D-Gel Electrophoresis in control and Nef-transfected SupT1 cells demonstrated several differentially expressed proteins with significant modulation of alpha-enolase. Through further studies, effects of Nef on alpha-enolase regulation were found to be cell lineage-specific, being stimulatory in macrophages/monocytes, inhibitory in T cells and without effect in HEK-293 cells. Cell migration and invasion studies were employed to determine biological function affected by Nef mediated regulation of alpha-enolase. Cell invasion was enhanced in THP-1 cells but was inhibited in SupT1 cells by wildtype nef. In addition, the modulation of enolase and cell invasion remained unaffected by a unique nef variant. These results indicated that regulation of alpha-enolase expression and invasive property of host cells by Nef is sequence specific, suggesting involvement of a particular motif of Nef. To precisely determine this site, we designed a heptapeptide including the suggested alpha-enolase regulating sequence of nef and a nef mutant with deletion of this site. Macrophages/monocytes being the major cells affected by HIV-1 at mucosal barriers, were particularly investigated by the nef mutant and peptide. Both the nef mutant and heptapeptide led to inhibition of enhanced enolase expression and increased invasiveness in THP-1 cells. Together, these findings suggest a possible mechanism of host invasion by HIV-1 through Nef mediated regulation of alpha-enolase and identifies a potential therapeutic target for HIV-1 entry at mucosal barriers.

Keywords: HIV-1 Nef, nef variants, host-virus interaction, tissue invasion

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Authors: Qasim A. El Dwairi, Saleh M. Banihani, Ayat S. Banihani, Ziad M. Bataineh

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Neurons in the spinal trigeminal nucleus of the camel were studied by Golgi impregnation. Neurons were classified based on differences in size and shape of their cell bodies, density of their dendritic trees, morphology and distribution of their appendages. In the spinal trigeminal nucleus of the camel, at least twelve types of neurons were identified. These neurons include, stalked, islets, octubus-like, lobulated, boat-like, pyramidal, multipolar, round, oval and elongated neurons. They have large number of different forms of appendages not only for their dendrites but also for their cell bodies. Neurons with unique large dilatations especially at their dendritic branching points were found. The morphological features of these neurons were described and compared with their counterparts in other species. Finding of large number of neuronal types with different size and shapes and large number of different forms of appendages for cell bodies and dendrites together with the presence of cells with unique features such as large dilated parts for dendrites may indicate to a very complex information processing for pain and temperature at the level of the spinal trigeminal nucleus in the camel that traditionally live in a very hard environment (the desert).

Keywords: camel, golgi, neurons , spinal trigeminal nucleus

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Authors: Mehdi Piri Ouchtapeh, Fariba Mehrkhou, Maryam Fourouzan

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The cabbage aphid, Bravicoryne brassicae (Hemiptera: Aphididae), is known as an economically important and oligophagous pest of different cole crops. The polyvolitine characteristics of B. brassicae resulted in resistance to insecticides. For this purpose, in this study, the sub-lethal concentration (LC25) of two insecticides, clothianidin and summer oil, on the life table parameters and population projection of cabbage aphid were studied at controlled condition (20±1 ℃, R.H. 60 ±5 % and a photoperiod of 16:8 h (L:D). The dipping method was used in bioassay and life table studies. Briefly, the leaves of cabbage containing 15 the same-aged (24h) adults of cabbage aphid (four replicates) were dipped into the related concentrations of insecticides for 10 s. The sub-lethal (LC25) obtained concentration were used 5.822 and 108.741 p.p.m for clothianidin and summer oil, respectively. The biological and life table studies were done using at least 100, 93 and 82 the same age of eggs for control, summer oil and clothianidin treatments respectively. The life history data of the greenhouse whitefly cohorts exposed to sublethal concentration of the aforementioned insecticides were analyzed using the computer program TWOSEX–MSChart based on the age-stage, two-sex life table theory. The results of this study showed that the used insecticides affected the developmental time, survival rate, adult longevity, and fecundity of the F1 generation. The developmental time on control, clothianidin and summer oil treatments was obtained (5.91 ± 0.10 days), (7.64 ± 0.12 days) and (6.66 ± 0.10 days), respectively. The sublethal concentration of clothianidin resulted in decreasing of adult longevity (8.63 ± 0.30 days), fecundity (14.14 ± 87 nymphs), survival rate (71%) and the life expectancy (10.26 days) of B. brassicae, as well. Additionally, usage of LC25 insecticides led to decreasing of the net reproductive rate (R0) of the cabbage aphid compared to summer oil and control treatments. The intrinsic rate of increase (r) (day-1) was decreased in F1 adults of cabbage aphid compared with other treatments. Additionally, the population projection results were accordance with the population growth rate of cabbage aphid. Therefore, the findings of this research showed that, however, both of the insecticides were effective on cabbage aphid population, but clothianidin was more effective and could be consider in the management of aforementioned pest.

Keywords: the cabbage aphid, sublethal effects, survival rate, population projection, life expectancy

Procedia PDF Downloads 79

Authors: Astari Indarsari, Bastian B. Purba, Muhammad Fadlilah

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In Indonesian climates that have the tropic climate, one of the potential energy sources is coming from solar energy. From the solar energy, we can convert it into the others energy, such as electrical energy. In this topic, we want to do the research about Dye Sensitized Solar Cell (DSSC). The materials that we use as sensitizer is anthocyanin that we extract from apple skin, because the anthocyanin is one of the most effective as a sensitizer for DSSC. The variable in this research is pH. The pH that we used are pH 0,5; pH 1; pH 1,5; pH 2; pH 2,5. The method is electrolysis, and we use TiO2 as sensitized material. The hypothesis from this research is the smaller pH can make higher the efficiency of the absorbent of the solar energy.

Keywords: anthocyanin, TiO2, DSSC, apple skin

Procedia PDF Downloads 293

Authors: Ihn Sook Jeong, Eun Joo Lee, Jae Hyung Kim, Gun Ho Kim, Young Jun Hwang

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This study investigated intravenous (IV) infiltration using bioelectrical impedance for 27 hospitalized patients in a long-term care hospital. Impedance parameters showed significant differences before and after infiltration as follows. First, the resistance (R) after infiltration significantly decreased compared to the initial resistance. This indicates that the IV solution flowing from the vein due to infiltration accumulates in the extracellular fluid (ECF). Second, the relative resistance at 50 kHz was 0.94 ± 0.07 in 9 subjects without infiltration and was 0.75 ± 0.12 in 18 subjects with infiltration. Third, the magnitude of the reactance (Xc) decreased after infiltration. This is because IV solution and blood components released from the vein tend to aggregate in the cell membrane (and acts analogously to the linear/parallel circuit), thereby increasing the capacitance (Cm) of the cell membrane and reducing the magnitude of reactance. Finally, the data points plotted in the R-Xc graph were distributed on the upper right before infiltration but on the lower left after infiltration. This indicates that the infiltration caused accumulation of fluid or blood components in the epidermal and subcutaneous tissues, resulting in reduced resistance and reactance, thereby lowering integrity of the cell membrane. Our findings suggest that bioelectrical impedance is an effective method for detection of infiltration in a noninvasive and quantitative manner.

Keywords: intravenous infiltration, impedance, parameters, resistance, reactance

Procedia PDF Downloads 182

Authors: Nabil A. Ahmed

Abstract:

DC-DC converters are necessary to interface low-voltage fuel cell power generation systems to a higher voltage DC bus system. A system and method for generating a regulated output power from fuel cell power generation systems is proposed in this paper, this includes a soft-switching isolated DC-DC converter to reduce the idling and circulating currents. The system incorporates a high-frequency center tap transformer link DC-DC converter using secondary-side soft switching control. Snubber capacitors including the parasitic capacitance of the switching devices and the transformer leakage inductance are utilized to achieve zero-voltage switching (ZVS) in the primary side of the high-frequency transformer. Therefore, no extra resonant components are required for ZVS. The inherent soft-switching capability allows high power density, efficient power conversion, and compact packaging. A prototype rated at 6.5 kW is proposed and simulated. Simulation results confirmed a wide range of soft-switching operation and consequently high conversion efficiency will be achieved.

Keywords: secondary-side, phase-shift, high-frequency transformer, zero voltage, zero current, soft switching operation, switching losses

Procedia PDF Downloads 310