Search results for: musculoskeletal graft versus host disease

Authors: Edilene Mercedes Mauer Machado, Carolina Hadassa Marques Karoly, Amanda Britz, Claudineia Brazil

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The El Niño phenomenon, characterized by the anomalous warming of surface waters in the Equatorial Pacific Ocean, can influence weather patterns in various parts of the world, including the occurrence of extreme events such as droughts or heavy rainfall. Studies have suggested a relationship between El Niño and an increase in the incidence of dengue in certain areas. During El Niño periods, there can be changes in climatic conditions, such as increased temperatures and reduced rainfall in certain tropical and subtropical regions. These conditions can favor the reproduction of the Aedes aegypti mosquito, the vector for dengue transmission. Research aims to investigate how climate events like El Niño and La Niña can influence the incidence and transmission of dengue. The results have shown that, on average, there was a significant increase in dengue cases during La Niña years compared to years of climatic neutrality, contradicting the findings of Hopp et al. (2015). The study also highlighted that regions affected by El Niño exhibited greater variability in dengue incidence. However, it is important to emphasize that the effects of El Niño on dengue transmission can vary depending on the region and local factors, such as socioeconomic context and implemented control measures, as described by Johansson et al. (2009). Not all areas affected by El Niño will necessarily experience an increase in dengue incidence, and the interaction between climate and disease transmission is complex.

Keywords: anomalous warming, climatic patterns, dengue incidence, extreme events

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Authors: Saimir Cenameri, Arjana Tereziu, Kastriot Dallaku

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Introduction: Intra-uterine death is a common problem in obstetrical practice, and can lead to complications if left to resolve spontaneously. The cervix is unprepared, making inducing of labor difficult. Misoprostol is a synthetic prostaglandin E1 analogue, inexpensive, and is presented valid thanks to its ability to bring about changes in the cervix that lead to the induction of uterine contractions. Misoprostol is quickly absorbed when taken orally, resulting in high initial peak serum concentrations compared with the vaginal route. The vaginal misoprostol peak serum concentration is not as high and demonstrates a more gradual serum concentration decline. This is associated with many benefits for the patient; fast induction of labor; smaller doses; and fewer side effects (dose-depended). Mostly it has been used the regime of 50 μg/4 hour, with a high percentage of success and limited side effects. Objective: Evaluation of the efficiency of the use of oral and vaginal misoprostol in inducing labor, and comparing it with its use not by a previously defined protocol. Methods: Participants in this study included patients at U.H.O.G. 'Koco Gliozheni', Tirana from April 2004-July 2006, presenting with an indication for inducing labor in the third trimester for pregnancy termination. A total of 37 patients were randomly admitted for birth inducing activity, according to protocol (26), oral or vaginal protocol (10 vs. 16), and a control group (11), not subject to the protocol, was created. Oral or vaginal misoprostol was administered at a dose of 50 μg/4 h, while the fourth group participants were treated individually by the members of the medical staff. The main result of interest was the time between induction of labor to birth. Kruskal-Wallis test was used to compare the average age, parity, women weight, gestational age, Bishop's score, the size of the uterus and weight of the fetus between the four groups in the study. The Fisher exact test was used to compare day-stay and causes in the four groups. Mann-Whitney test was used to compare the time of the expulsion and the number of doses between oral and vaginal group. For all statistical tests used, the value of P ≤ 0.05 was considered statistically significant. Results: The four groups were comparable with regard to woman age and weight, parity, abortion indication, Bishop's score, fetal weight and the gestational age. There was significant difference in the percentage of deliveries within 24 hours. The average time from induction to birth per route (vaginal, oral, according to protocol and not according to the protocol) was respectively; 10.43h; 21.10h; 15.77h, 21.57h. There was no difference in maternal complications in groups. Conclusions: Use of vaginal misoprostol for inducing labor in the third trimester for termination of pregnancy appears to be more effective than the oral route, and even more to uses not according to the protocols approved before, where complications are greater and unjustified.

Keywords: inducing labor, misoprostol, pregnancy termination, third trimester

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Authors: Omnia M. Kandil, Noha M. F. Hassan, Doaa Sedky, Hatem A. Shalaby, Heba M. Ashry, Nadia M. T. Abu El Ezz, Sahar M. Kandeel, Mohamed S. Abdelfattah Ying L, Ebtesam M. Al-Olayan

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The aim of the present study was to determine the prevalence of bovine cysticercosis in both cattle and buffaloes in Egypt and to assess the cysticidal efficacy of Balanites aegyptiacafruits (B. aegyptiaca) and Moringa oleiferaseeds (M. oleifera) extracts in experimentally infected mice. The study detected the level of tumor necrosis factor (TNF-α) to monitor the immune and inflammatory responses of experimentally infected mice. Through meat inspection, a total number of 2125 male bovines, 2 to 5 years old (1125 cattle and 1000 buffaloes) were examined at official abattoirs in Cairo Governorate. The prevalence of the disease among bovine was 7.8%, (6.31% of cattle and 9.5% of buffaloes). A decrease in the number of cysticerci was found in all treated mice groups, and up to 88% reduction was achieved in the B. aegyptiaca-treated group; higher than that was recorded in both M. oleifera (72.23%) and albendazole-treated ones (80.56%). Postmortem findings proved that M. oleifera and B. aegyptiaca reduced cysticerci numbers comparable to a commercial anthelmintic. The study showed a significant decrease (P<0.001) in TNF-α levels after treatment with Balanites and Moringa extracts, compared with the untreated control and the albendazole-treated groups.

Keywords: prevalence, bovine cysticercossis, extracts, mice

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Authors: Anika Pallapothu

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Periodontitis, a severe gum disease, poses a significant threat to the integrity of bone and soft tissues supporting teeth, primarily initiated by bacterial accumulation around the gum line. Conventional treatments like scaling/root planning and plaque removal are widely employed, but integrating modern technologies such as nanotechnology holds promise for innovative therapeutic approaches. This study explores the utilization of silver nanoparticles encapsulated within cellulose nanofiber (CNF) and mesoporous bioactive glass hydrogel matrices for periodontitis management. Silver nanoparticles exhibit potent antimicrobial properties by disrupting microbial cell membranes, inducing reactive oxygen species (ROS) generation, and interfering with vital cellular processes like ATP production and nucleic acid synthesis. Mesoporous bioactive glass, renowned for its high surface area, osteoconductive, and bioactivity, presents a favorable platform for pharmaceutical applications. Incorporating CNF enhances the properties of the hydrogel due to its biocompatibility, biodegradability, and water absorption capacity. The proposed composite material is anticipated to exert beneficial effects in periodontitis treatment by demonstrating antibacterial and anti-inflammatory activities, offering a promising avenue for future therapeutic interventions.

Keywords: periodontitis, cellulose nanofibers, silver nanoparticles, mesoporous bioactive glass, antibacterial activity, anti-inflammatory activity

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Authors: Arati Inamdar, Siddharth Bhattacharyya, Kester Haye

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Collision tumors are rare entities characterized by neoplasms of two different cell populations with distinct separating boundaries. Such tumors could be benign, malignant, or a combination of both. The exact mechanism of origin for collision tumors is predicted to be tumor heterogeneity or concurrent occurrence of neoplasm in the same organ. We present two cases of plasmacytoma presenting as a collision tumor, one with a tumor of hematological origin and another with a non-hematological origin, namely Chronic Lymphocytic Leukemia and Adenocarcinoma of the colon, respectively. The immunohistochemical stains and flowcytometry analysis performed on the specimens aided incorrect diagnosis. Interestingly, neoplastic cells of plasmacytoma in the first case demonstrated strong cytokeratin along with weak Epithelial Specific Antigen/ Epithelial cell adhesion molecule Monoclonal Antibody (MOC31) positivity, indicating that the tumor may influence the microenvironment of the tumor in the vicinity. Furthermore, the next-generation sequencing studies performed on the specimen with plasmacytoma and chronic lymphocytic lymphoma demonstrated BReast CAncer gene (BRCA2) and Tumor Necrosis Factor Alpha Induced Protein 3 (TNFAIP3) as a disease associated variants suggestive of risk for multiple tumors including collision tumors. Our reports highlight the unique collision tumors involving plasmacytoma, which have never been reported previously, as well as provide necessary insights about the underline genetic aberrations and tumor heterogeneity through sequencing studies and allow clonality assessment for subsequent tumors.

Keywords: BRCA2, collision tumor, chronic lymphocytic leukemia, plasmacytoma

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Authors: Mehdi Nasr-Esfahani, Leila Mohammad Bagheri, Ava Nasr-Esfahani

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Root and collar rot disease caused by Phytophthora capsici (Leonian) is one of the most serious diseases in pepper, Capsicum annuum L. In this study, a diverse collection of 37 commercial edible and ornamental pepper genotypes infected with P. capsici were investigated for biomass parameters and enzymatic activity of peroxidase or peroxide reductases (EC), superoxide dismutase (SOD), polyphenol oxidase (PPOs), catalase (CAT) and phenylalanine ammonia-lyase (PAL). Seven candidate DEG genes were also evaluated on resistant and susceptible pepper cultivars, through measuring product formation, using spectrophotometry and real-time polymerase chain reaction. All the five enzymes and seven defense-gene candidates were up-regulated in all inoculated pepper accessions to P. capsici. But, the enzymes and DEG genes were highly expressed in resistant cv. 19OrnP-PBI, 37ChillP-Paleo, and “23CherryP-Orsh". The expression level of enzymes were 1.5 to 5.6-fold higher in the resistant peppers, than the control non-inoculated genotypes. Also, the transcriptional levels of related candidate DEG genes were 3.16 to 5.90-fold higher in the resistant genotypes. There was a direct and high correlation coefficient between resistance, bio-mass parameters, enzymatic activity, and resistance gene expression. The related enzymes and candidate genes expressed herein will provide a basis for further gene cloning and functional verification studies, and also will aid in an understanding of the regulatory mechanism of pepper resistance to P. capsici.

Keywords: AP2/ERF, cDNA, enzymes, MIP gene, q-RTPCR, XLOC

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Authors: Amirreza Razzaghipour, Mahdi Khalili

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References from the World Health Organization and the Center for Disease Control for deceleration the spread of the Novel COVID-19, comprises social estrangement, frequent handwashing, and covering your mouth when around others. As hearing healthcare specialists, the influence of existenceinvoluntary to boundary social interactions on persons with hearing impairment was significant for us to understand. We found ourselves delaying cochlear implant (CI) surgeries. All children, and chiefly those with hearing loss, are susceptible to reductions in spoken communication. Hearing plans, such as cochlear implants, provide children with hearing loss access to spoken communication and provision language development. when provided early and used consistently, these supplies help children with hearing loss to engage in spoken connections. Cochlear implant (CI) is a standard medical-surgical treatment for bilateral severe to profound hearing loss with no advantage with the hearing aid. Hearing is one of the most important senses in humans. Pediatric hearing loss establishes one of the most important public health challenges. Children with hearing loss are recognized early and habilitated via hearing aids or with cochlear implants (CIs). Suitable care and maintenance as well as continuous auditory verbal therapy (AVT) are also essential in reaching for the successful attainment of language acquisition. Children with hearing loss posture important challenges to their parents, particularly when there is limited admission to their hearing care providers. The disruption in the routine of their hearing and therapy follow-up services has had substantial effects on the children as well as their parents.

Keywords: healthcare, covid-19, cochlear implants, spoken communication, hearing loss

Procedia PDF Downloads 153

Authors: Chakrit Thapphan, Suteeraporn Chaowattanapanit, Sorutsiri Chareonsudjai, Wisitsak Phoksawat, Supranee Phantanawiboon, Kiatichai Faksri, Steve W. Edwards, Kanin Salao

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Background: Psoriasis is a chronic inflammatory disease of the skin that is mediated by crosstalk between keratinocytes and immune cells, especially CD4+ T helper (Th) cells. To date, psoriasis is established as a T helper 17 (Th17) cell-mediated inflammatory process driven by the over-expression of Th17. However, the role of other CD4+T helper cells is rather controversial. Objective: Our study, thereby, aimed to characterize and analyze T cell subsets in the circulating blood of psoriasis patients and compare them to healthy controls. Methods: Peripheral blood mononuclear cells were isolated from the participants and stained with fluorescent dye-conjugated monoclonal antibodies specific for intracellular cytokines, including interferon-gamma (IFN- γ), interleukin (IL-4), IL-17 and forkhead box P3 (FOXP3), that can be used to define T helper 1 (Th1) cells, T helper 2 (Th2), T helper 17 (Th17) and regulatory T cells (Treg) respectively. Results: We found that the numbers of Th2 (59.6% ± 17.0) and Th17 (4.0% ± 2.0) cells in the circulating blood of psoriasis patients were significantly higher than those of the healthy controls (p= 0.0007 and 0.0013 respectively). In contrast, the numbers of Th1 and Treg cells were not significantly different between psoriasis patients and healthy controls (p= 0.0593 and 0.8518, respectively). Additionally, when adjusting these numbers of Th cells to Treg, we observed a similar trend that the ratio of Th2/Treg and Th17/Treg also elevated (p = 0.0007 and 0.0047, respectively). Conclusion: Taken together, our results suggest an imbalanced T exhibit toward the Th2 and Th17 skewed-immune responses in psoriasis patients.

Keywords: psoriasis, Th cell subsets, Th2 cells, Th17 cells, Treg cells

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Authors: Summer Hassan, David Crossman

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Myocardial fibrosis (MF) has been loosely defined as the process occurring in the pathological remodeling of the myocardium due to excessive production and deposition of extracellular matrix (ECM) proteins, including collagen. This reduces tissue compliance and accelerates progression to heart failure, as well as affecting the electrical properties of the myocytes resulting in arrhythmias. Microscopic interrogation of MF is key to understanding the molecular orchestrators of disease. It is well-established that recruitment and stimulation of myofibroblasts result in Collagen deposition and the resulting expansion in the ECM. Many types of Collagens have been identified and implicated in scarring of tissue. In a series of experiments conducted at our lab, we aim to elucidate the role collagen VI plays in the development of myocardial fibrosis and its direct impact on myocardial function. This was investigated through an animal experiment in Rats with Collagen VI knockout diseased and healthy animals as well as Collagen VI wild diseased and healthy rats. Echocardiogram assessments of these rats ensued at four-time points, followed by microscopic interrogation of the myocardium aiming to correlate the role collagen VI plays in myocardial function. Our results demonstrate a deterioration in cardiac function as represented by the ejection fraction in the knockout healthy and diseased rats. This elucidates a potential protective role that collagen-VI plays following a myocardial insult. Current work is dedicated to the microscopic characterisation of the fibrotic process in all rat groups, with the results to follow.

Keywords: heart failure, myocardial fibrosis, collagen, echocardiogram, confocal microscopy

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Authors: Navkiran Kaur, Apoorva Mathur, Abhishree Agarwal, Sakshi Gupta, Tuhin Rashmi

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Aim: Breast cancer is the most common cancer in women worldwide, and resistance to the current therapeutics, often concurrently, is an increasing clinical challenge. Glycosylation of proteins is one of the most important post-translational modifications. It is widely known that aberrant glycosylation has been implicated in many different diseases due to changes associated with biological function and protein folding. Alterations in cell surface glycosylation, can promote invasive behavior of tumor cells that ultimately lead to the progression of cancer. In breast cancer, there is an increasing evidence pertaining to the role of glycosylation in tumor formation and metastasis. In the present study, an attempt has been made to study the disease associated sialoglycoproteins in breast cancer by using bioinformatics tools. The sequence will be retrieved from UniProt database. A database in the form of a word document was made by a collection of FASTA sequences of breast cancer gene sequence. Glycosylation was studied using yinOyang tool on ExPASy and Differential genes expression and protein analysis was done in context of breast cancer metastasis. The number of residues predicted O-glc NAc threshold containing 50 aberrant glycosylation sites or more was detected and recorded for individual sequence. We found that the there is a significant change in the expression profiling of glycosylation patterns of various proteins associated with breast cancer. Differential aberrant glycosylated proteins in breast cancer cells with respect to non-neoplastic cells are an important factor for the overall progression and development of cancer.

Keywords: breast cancer, bioinformatics, cancer, metastasis, glycosylation

Procedia PDF Downloads 280

Authors: W. Phokhwang-Just, A. Saraketrin, P. Thongpet, J. Udomkitpipat, J. Kaewsakulthong

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The more people get older and live longer, the more health problems they may have. This cross-sectional study aims to study a correlation between death anxiety, quality of life, and self-esteem as well as factors affecting these variables in the elderly living in Surat Thani Province, Thailand. Of 382 elderly people, who were proportionally sampled from 19 districts in Surat Thani Province, 256 (67%) already returned the questionnaires. The Thai version of Templer’s Death Anxiety, Quality of Life (WHO-BREF), and of Rosenberg’s Self-Esteem Questionnaires were employed. The result showed that the samples had a mean age of 72 years old, 53% were female, 62% were married, 61% graduated with primary-school, and 61% had at least one chronic disease Approximately, 19% of them had 3 diseases. The quality of life (QOL), self-esteem (SE), and death anxiety (DA) of samples were in moderate (n= 91, mean = 86.89, SD = 15.47), high (n = 138, mean = 29.33, SD=4.77), and low level (n= 85, mean = 6.23, SD= 3.65), respectively. The QOL was not significantly different between male and female as well as among different marital status. The female elderly had more DA and less SE than male (t= 2.095, df = 83; t =-3.258, df =135, respectively, p < 0.05). The female elderly, who were separated or widow, had a higher level of DA than did the married elderly (LSD: p < 0.05). The married elderly had a higher level of SE than did the separated, widowed (Tukey HSD, LSD: p < 0.05), or single elderly (LSD: p < 0.05). The more diseases the elderly got, the lower level of QOL they had (r = -0.335, p < 0.05). The QOL was significantly correlated with SE (r =0.434, p < 0.05), but not significantly related to DA (r = -0.200, p = 0.069). The lower level of SE the elderly had, the higher level of DA they become (r = -2.71, p < 0.05). In order to promote the QOL, the SE of the elderly should be enhanced. Consequently, the DA can be minimized. Healthcare providers should provide care that promotes QOL, SE, and reduces DA of the elderly, especially those, who are female, single, and separated or widowed as well as those, who have more diseases than the others

Keywords: death anxiety, quality of life, self-esteem, elderly

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Authors: Yin Hua Zhang

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Cardiac metabolism is essential in myocardial contraction. In addition to glucose, fatty acids (FA) are essential in producing energy in the myocardium since FA-dependent beta-oxidation accounts for > 70-90% of cellular ATP under resting conditions. However, metabolism shifts from FAs to glucose utilization during disease progression (e.g. hypertrophy and ischemic myocardium), where glucose oxidation and glycolysis become the predominant sources of cellular ATP. At advanced failing stage, both glycolysis and beta-oxidation are dysregulated, result in insufficient supply of intracellular ATP and weakened myocardial contractility. Undeniably, our understandings of myocyte function in healthy and diseased hearts are based on glucose (10 mM)-dependent metabolism because glucose is the “sole” metabolic substrate in most of the physiological experiments. In view of the importance of FAs in cardiovascular health and diseases, we aimed to elucidate the impacts of FA supplementation on myocyte contractility and evaluate cellular mechanisms those mediate the functions in normal heart and with pathological stress. In particular, we have investigated cardiac excitation-contraction (E-C) coupling in the presence and absence of FAs in normal and hypertensive rat left ventricular (LV) myocytes. Our results reveal that FAs increase mitochondrial activity, intracellular [Ca²+]i, and LV myocyte contraction in healthy LV myocytes, whereas FA-dependent cardiac inotropyis attenuated in hypertension. FA-dependent myofilament Ca²+ desensitization could be fundamental in regulating [Ca²+]i. Collectively, FAs supplementation resets cardiac E-C coupling scheme in healthy and diseased hearts.

Keywords: hypertension, fatty acid, heart, calcium

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Authors: Amruta D. S. Pathare, Indira Hinduja, Kusum Zaveri

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Implantation failure (IF) even after the good-quality embryo transfer (ET) in the physiologically normal endometrium is the main obstacle in in vitro fertilization (IVF). Various microarray studies have been performed worldwide to elucidate the genes requisite for endometrial receptivity. These studies have included the population based on different phases of menstrual cycle during natural cycle and stimulated cycle in normal fertile women. Additionally, the literature is also available in recurrent implantation failure patients versus oocyte donors in natural cycle. However, for the first time, we aim to study the genomics of endometrial receptivity in IF patients under controlled ovarian stimulation (COS) during which ET is generally practised in IVF. Endometrial gene expression profiling in IF patients (n=10) and oocyte donors (n=8) were compared during window of implantation under COS by whole genome microarray (using Illumina platform). Enrichment analysis of microarray data was performed to determine dys-regulated biological functions and pathways using Database for Annotation, Visualization and Integrated Discovery, v6.8 (DAVID). The enrichment mapping was performed with the help of Cytoscape software. Microarray results were validated by real-time PCR. Localization of genes related to immune response (Progestagen-Associated Endometrial Protein (PAEP), Leukaemia Inhibitory Factor (LIF), Interleukin-6 Signal Transducer (IL6ST) was detected by immunohistochemistry. The study revealed 418 genes downregulated and 519 genes upregulated in IF patients compared to healthy fertile controls. The gene ontology, pathway analysis and enrichment mapping revealed significant downregulation in activation and regulation of immune and inflammation response in IF patients under COS. The lower expression of Progestagen Associated Endometrial Protein (PAEP), Leukemia Inhibitory Factor (LIF) and Interleukin 6 Signal Transducer (IL6ST) in cases compared to controls by real time and immunohistochemistry suggests the functional importance of these genes. The study was proved useful to uncover the probable reason of implantation failure being imbalance of immune and inflammatory regulation in our group of subjects. Based on the present study findings, a panel of significant dysregulated genes related to immune and inflammatory pathways needs to be further substantiated in larger cohort in natural as well as stimulated cycle. Upon which these genes could be screened in IF patients during window of implantation (WOI) before going for embryo transfer or any other immunological treatment. This would help to estimate the regulation of specific immune response during WOI in a patient. The appropriate treatment of either activation of immune response or suppression of immune response can be then attempted in IF patients to enhance the receptivity of endometrium.

Keywords: endometrial receptivity, immune and inflammatory response, gene expression microarray, window of implantation

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Authors: Djoudi Abdelhak, Djibaoui Rachid, Reguieg Yassaad Houcine

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Our study focuses on the identification of some species of Pseudomonas (P4, P5, P7 and P8) isolated from saline soils in northwestern Algeria and the effect of their metabolites on the growth of Alternaria alternata the causative agent of the blight of the bean disease (Vicia faba). We are also interested in stimulating the growth of this plant species in saline conditions (60 mM/l NaCl) and the absence of salts. The analysis focuses on rates of inhibition of mycelial growth of Alternaria alternata strain and the rate of growth of plants inoculated with strains of Pseudomonas expressed by biometrics. According to the results of the in-vitro test, P5 and P8 species and their metabolites showed a significant effect on mycelia growth and production of spores of Alternaria alternata. The in-vivo test shows that the species P8 and P5 were significantly and positively influencing the growth in biometric parameters of the bean in saline and salt-free condition. Inoculation with strain P5 has promoted the growth of the bean in stem height, stem fresh weight and dry weight of stems of 108.59%, 115.28%, 104.33%, respectively, in the presence of salt Inoculation with strain P5 has fostered the growth of the bean stem fresh weight of 112.47% in the presence of salt The effect of Pseudomonas species on the development of Vicia faba and the growth of Alternaria alternata is considering new techniques and methods of biological production and crop protection.

Keywords: pseudomonas, vicia faba, alternaria alternata, promoting of plant growth

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Authors: K. P. Ganesh, T. Gomathi, L. Arul Pragasan

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Degradation of lake water quality is one of the serious environmental threats for the last few decades, particularly, the lakes situated in and around urban and industrial areas. The present study aimed to analyze the physicochemical and biological parameters, and metal elements to determine the water quality of Krishnampathi, Ukkadam, Kurichi, Sulur and Singanallur Lakes. Of the 23 physicochemical parameters analyzed in the five lakes, except TDS, Chloride and Total hardness values all the 20 parameters were found within the prescribed limit as recommended by World Health Organization (WHO) and Bureau of Indian Standards (BIS). In case of biological parameter, both Total Coliform and Fecal Coliform bacteria (Escherichia coli) were identified. This indicates the contamination of lakes by fecal matter, and warns of potential of disease causing by viruses, bacteria and other organisms. Among the twelve metal elements (Al, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Mo, Cd and Pb) determined by inductively coupled plasma-mass spectroscopy, except Cd (for all lakes), and Pb (for Ukkadam, Kurichi, Sulur & Singanallur), all the elements were found above the prescribed limits of BIS. The results of the present study revealed that all the five major lakes of Coimbatore were contaminated. It is recommended that proper implementation of the new wetland waste management system and monitoring of water quality be of the urgent need to sustain the water bodies for future generations.

Keywords: heavy metals, inductively coupled plasma-mass spectroscopy, physicochemical and biological parameters, water quality

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Authors: Asha Manoharan, Sooraj A. O, Anju K. G

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Enlarged vestibular aqueduct (EVA) refers to the presence of congenital sensorineural hearing loss with an enlarged vestibular aqueduct. The Audiological symptoms of EVA are fluctuating and progressive in nature and the diagnosis of EVAS can be confirmed only with radiological evaluation. Hence it is difficult to differentiate EVA from conditions like Meniere’s disease, semi-circular dehiscence, etc based on audiological findings alone. EVA in adults is easy to identify due to distinct vestibular symptoms. In children, EVA can remain either unidentified or misdiagnosed until the vestibular symptoms are evident. Motor developmental delay, especially the ones involving a change of body alignment, has been reported in the pediatric population with EVA. So, it should be made mandatory to recommend radiological evaluation in young children with fluctuating hearing loss reporting with motor developmental delay. This single case study of a baby with Enlarged Vestibular Aqueduct (EVA) primarily aimed to address the following: a) Challenges while diagnosing young patients with EVA and fluctuating hearing loss, b) Importance of radiological evaluation in audiological diagnosis in the pediatric population, c) Need for regular monitoring of hearing, hearing aid performance, and cochlear implant mapping closely for potential fluctuations in such populations, d) Importance of reviewing developmental, language milestones in very young children with fluctuating hearing loss.

Keywords: enlarged vestibular aqueduct (EVA), motor delay, radiological evaluation, fluctuating hearing loss, cochlear implant

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Authors: Mustafa M. Donma, Orkide Donma

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Iron is physiologically essential. However, it also participates in the catalysis of free radical formation reactions. Its deficiency is associated with amplified health risks. This trace element establishes some links with another physiological process related to cell death, apoptosis. Both iron deficiency and iron overload are closely associated with apoptosis. Soluble tumor necrosis factor-like weak inducer of apoptosis (sTWEAK) has the ability to trigger apoptosis and plays a dual role in the physiological versus pathological inflammatory responses of tissues. The aim of this study was to investigate the status of these parameters as well as the associations among them in children with obesity, a low-grade inflammatory state. The study was performed on groups of children with normal body mass index (N-BMI) and obesity. Forty-three children were included in each group. Based upon age- and sex-adjusted BMI percentile tables prepared by World Health Organization, children whose values varied between 85 and 15 were included in N-BMI group. Children whose BMI percentile values were between 99 and 95 comprised obese (OB) group. Institutional ethical committee approval and informed consent forms were taken prior to the study. Anthropometric measurements (weight, height, waist circumference, hip circumference, head circumference, neck circumference) and blood pressure values (systolic blood pressure and diastolic blood pressure) were recorded. Routine biochemical analysis including serum iron, total iron binding capacity (TIBC), transferrin saturation percent (Tf Sat %), and ferritin were performed. Soluble tumor necrosis factor-like weak inducer of apoptosis levels were determined by enzyme-linked immunosorbent assay. Study data was evaluated using appropriate statistical tests performed by the statistical program SPSS. Serum iron levels were 91±34 mcrg/dl and 75±31 mcrg/dl in N-BMI and OB children, respectively. The corresponding values for TIBC, Tf Sat %, ferritin were 265 mcrg/dl vs 299 mcrg/dl, 37.2±19.1 % vs 26.7±14.6 %, and 41±25 ng/ml vs 44±26 ng/ml. in N-BMI and OB groups, sTWEAK concentrations were measured as 351 ng/L and 325 ng/L, respectively (p>0.05). Correlation analysis revealed significant associations between sTWEAK levels and iron related parameters (p<0.05) except ferritin. In conclusion, iron contributes to apoptosis. Children with iron deficiency have decreased apoptosis rate in comparison with that of healthy children. sTWEAK is inducer of apoptosis. Obese children had lower levels of both iron and sTWEAK. Low levels of sTWEAK are associated with several types of cancers and poor survival. Although iron deficiency state was not observed in this study, the correlations detected between decreased sTWEAK and decreased iron as well as Tf Sat % values were valuable findings, which point out decreased apoptosis. This may induce a proinflammatory state, potentially leading to malignancies in the future lives of obese children.

Keywords: apoptosis, children, iron-related parameters, obesity, soluble tumor necrosis factor-like weak inducer of apoptosis

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Authors: Akinbo Razak Yinka, Adesanya Kehinde Kazeem, Oladokun Oluwagbenga Peter

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Time series data are sequences of observations collected over a period of time. Such data can be used to predict health outcomes, such as disease progression, mortality, hospitalization, etc. The Statistical approach is based on mathematical models that capture the patterns and trends of the data, such as autocorrelation, seasonality, and noise, while Neural methods are based on artificial neural networks, which are computational models that mimic the structure and function of biological neurons. This paper compared both parametric and non-parametric time series models of patients treated for malaria in Maternal and Child Health Centres in Lagos State, Nigeria. The forecast methods considered linear regression, Integrated Moving Average, ARIMA and SARIMA Modeling for the parametric approach, while Multilayer Perceptron (MLP) and Long Short-Term Memory (LSTM) Network were used for the non-parametric model. The performance of each method is evaluated using the Mean Absolute Error (MAE), R-squared (R2) and Root Mean Square Error (RMSE) as criteria to determine the accuracy of each model. The study revealed that the best performance in terms of error was found in MLP, followed by the LSTM and ARIMA models. In addition, the Bootstrap Aggregating technique was used to make robust forecasts when there are uncertainties in the data.

Keywords: ARIMA, bootstrap aggregation, MLP, LSTM, SARIMA, time-series analysis

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Authors: Anjani Kumar Tiwari, Neelam Kumari, Anil Mishra

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The family of phosphodiesterases (PDEs) plays a critical role in control of the level, localization, and duration of intracellular 3’-5’-cyclic adenosine monophosphate (cAMP) and 3’-5’-cyclic guanosine monophosphate (cGMP) signals by specifically hydrolyzing these cyclic nucleotides. As the involvement of cyclic nucleotide second messengers in cell signaling and homeostasis is established, the regulation of these pathways in the brain by various PDE isoforms is an area of considerable interest, as they are involved in nearly all brain functions and in the etiology of neuropsychiatric diseases. The PDE10A isoform, isolated from different species and characterized regarding structure and function, has received much attention in recent years, particularly in the context of schizophrenia and Huntington’s disease, which are both related to a role of PDE10A in the regulation of striatal dopaminergic neurotransmission. Quinazoline analogue 1-(4-methoxyphenyl)-6,7-dimethoxyquinazoline, was evaluated as specific PET marker for phosphodiesterase (PDE) 10A. Here, we report the radiosynthesis of [11C]2 and the in vitro and in vivo evaluation of [11C]2 as a potential positron emission tomography (PET) radiotracer for imaging PDE10A in the central nervous system (CNS). The radiosynthesis of [11C]2 was achieved by O-methylation of the corresponding des-methyl precursor with [11C]methyl iodide. [11C]2 was obtained with ∼50% radiochemical yield. PET imaging studies in rat brain displayed initial specific uptake with very rapid clearance of [11C]2 from brain. Though [11C]2 is not an ideal radioligand for clinical imaging of PDE10A in the CNS. Modified analogue of quinazoline having a higher potency for inhibiting PDE10A and improved pharmacokinetic properties will be necessary for imaging this enzyme with PET.

Keywords: PDE10A, PET, radiotracer, quinazoline

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Authors: Rudzani Manafe, Ezekiel Green

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Brucella, has many different virulence factors that act as a causative agent of brucellosis, depending on the environment and other factors, some factors may play a role more than others during infection and as a result, play a role in becoming a causative agent for pathogenesis. Brucella melitensis and Brucella abortus are considered to be pathogenic to humans. The genetic regularity of nine potential causes of virulence of two Brucella species in Eastern Cape livestock have been examined. A hundred and twenty isolates obtained from Molecular Pathogenesis and Molecular Epidemiology Research Group (MPMERG) were used for this study. All isolates were grown on Brucella agar medium. Nine primer pairs were used for the detection of virB2, virB5, vceC, btpA, btpB, prpA, betB, bpe275, and bspB virulence factors using Polymerase chain reaction (PCR). Approximately 100% was observed for genes BecC and BetB from B. arbotus. While the lowest gene observed was PrpA at 4.6% from B. arbotus. BetB was detected in 34.7%, while virB2 and prpA (0%) were not detected in B. melitensis. The results from this research suggest that most isolates of Brucella have virulence-related genes associated with disease pathogenesis. Finally, our findings showed that Brucella strains in the Eastern Cape Province are extremely virulent as virulence characteristics exist in most strains investigated.

Keywords: putative virulence genes, brucella, polymerase chain reaction, milk

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Authors: A. J. Olatunji, Adebolu Temitope Johnson

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Groundwater samples were collected randomly from hand-dug wells and boreholes in parts of the Ado Ekiti metropolis and were subjected to quality assessment and characterization. Physicochemical analyses, which include the in-situ parameters (pH units, Turbidity, and Electrical Conductivity) and laboratory analysis of selected ionic concentrations, were carried out following standard methods. Hydrochemistry of the present study revealed relative mean concentrations of cations in the order Ca2+ > Na+ > Mg2+ > Cu2+> Fe > Mn2+ and that of anions: Cl- > NO3- > SO42- > F - respectively considering World Health Organisation Standard (WHO) range of values for potable water. The result shows that values of certain parameters (Total Dissolved Solid (TDS), Manganese, Calcium, Magnesium, Fluoride, and Sulphate) were below the Highest Desirable Level of the Standards, while values of some other parameters (pH Units, Electrical Conductivity, Turbidity, Alkalinity, Sodium, Copper, Chloride, and Total Hardness) were within the range of figures between Highest Desirable Level (HDL) and Maximum Permissible Level (MPL) of World Health Organization (WHO) drinking water Standards. The reduction in the mean concentration value of Total Dissolved Solids (TDS) of most borehole samples follows the fact that water had been allowed to settle in the overhead tanks before usage; we discussed and brainstormed in the course of sampling and agreed to take a sample that way because that represents what the people consume, it also shows an indication while there was slightly concentration increase of these soluble ions in hand-dug wells samples than borehole samples only with the exception of borehole sample seven BH7 because BH7 uses the mono-pumping system. These in-situ parameters and ionic concentrations were further displayed and or represented on bar charts along with the WHO standards for better pictorial clarifications. Deductions from field observation indices revealed the imprints of natural weathering, ion-exchange processes, and anthropogenic activities influencing groundwater quality. A strong degree of association was found to exist between sodium and chlorine ions in both hand-dug well and borehole groundwater samples through the use of Pearson’s correlation coefficient; this association can further be supported by the chemistry of the parent bedrock associated with the study area because the chemistry of groundwater is a replica of its host rock. The correlation of those two ions must have begun from the period of mountain building, indicating an identical source from which they were released to the groundwater. Moreover, considering the comparison of ionic species concentrations of all samples with the (WHO) standards, there were no anomalous increases or decreases in the laboratory analysis results; this simply reveals an insignificant state of pollution of the groundwater. The study and its sampling techniques were not set to target the likely area and extent of groundwater pollution but its portability. It could be said that the samples were safe for human consumption.

Keywords: groundwater, physicochemical, parameters ionic, concentrations, WHO standards

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Authors: Shiva Naidoo, Kristena Yossef, Grimm Jimm, Mirza Wasique, Eric Kemmerer, Joshua Obuch, Anand Mahadevan

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Background: Fiducial markers effectively enhance tumor target visibility prior to Stereotactic Body Radiation Therapy or Proton therapy. To streamline clinical practice, fiducial placement guidelines from a robotic radiosurgery vendor were examined with the goals of optimizing and standardizing feasible geometries for each treatment indication. Clinical examples of prostate, renal, and pancreatic cases are presented. Methods: Vendor guidelines (Accuray, Sunnyvale, Ca) suggest implantation of 4–6 fiducials at least 20 mm apart, with at least a 15-degree angular difference between fiducials, within 50 mm or less from the target centroid, to ensure that any potential fiducial motion (e.g., from respiration or abdominal/pelvic pressures) will mimic target motion. Also recommended is that all fiducials can be seen in 45-degree oblique views with no overlap to coincide with the robotic radiosurgery imaging planes. For the prostate, a standardized geometry that meets all these objectives is a 2 cm-by-2 cm square in the coronal plane. The transperineal implant of two pairs of preloaded tandem fiducials makes the 2 cm-by-2 cm square geometry clinically feasible. This technique may be applied for renal cancer, except repositioned in a sagittal plane, with the retroperitoneal placement of the fiducials into the tumor. Pancreatic fiducial placement via endoscopic ultrasound (EUS) is technically more challenging, as fiducial placement is operator-dependent, and lesion access may be limited by adjacent vasculature, tumor location, or restricted mobility of the EUS probe in the duodenum. Fluoroscopically assisted fiducial placement during EUS can help ensure fiducial markers are deployed with optimal geometry and visualization. Results: Among the first 22 fiducial cases on a newly installed robotic radiosurgery system, live x-ray images for all nine prostatic cases had excellent fiducial visualization at the treatment console. Renal and pancreatic fiducials were not as clearly visible due to difficult target access and smaller caliber insertion needle/fiducial usage. The geometry of the first prostate case was used to ensure accurate geometric marker placement for the remaining 8 cases. Initially, some of the renal and pancreatic fiducials were closer than the 20 mm recommendation, and interactive feedback with the proceduralists led to subsequent fiducials being too far to the edge of the tumor. Further feedback and discussion of all cases are being used to help guide standardized geometries and achieve ideal fiducial placement. Conclusion: The ideal tradeoffs of fiducial visibility versus the thinnest possible gauge needle to avoid complications needs to be systematically optimized among all patients, particularly in regards to body habitus. Multidisciplinary collaboration among proceduralists and radiation oncologists can lead to improved outcomes.

Keywords: fiducial, prostate cancer, renal cancer, pancreatic cancer, radiotherapy

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Authors: Philippe Gugler

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Goal of the study: The concept of competitiveness has been addressed by economic theorists and policymakers for several hundreds of years, with both groups trying to understand the drivers of economic prosperity and social welfare. The goal of this contribution is to address the major useful theoretical contributions that permit to identify the main drivers of a territory’s competitiveness. We first present the major contributions found in the classical and neo-classical theories. Then, we concentrate on two majors schools providing significant thoughts on the competitiveness of locations: the Economic Geography (EG) School and the International Business (IB) School. Methodology: The study is based on a literature review of the classical and neo-classical theories, on the economic geography theories and on the international business theories. This literature review establishes links between these theoretical mainstreams. This work is based on the academic framework establishing a meaningful literature review aimed to respond to our research question and to develop further research in this field. Results: The classical and neo-classical pioneering theories provide initial insights that territories are different and that these differences explain the discrepancies in their levels of prosperity and standards of living. These theories emphasized different factors impacting the level and the growth of productivity in a given area and therefore the degree of their competitiveness. However, these theories are not sufficient to more precisely identify the drivers and enablers of location competitiveness and to explain, in particular, the factors that drive the creation of economic activities, the expansion of economic activities, the creation of new firms and the attraction of foreign firms. Prosperity is due to economic activities created by firms. Therefore, we need more theoretical insights to scrutinize the competitive advantages of territories or, in other words, their ability to offer the best conditions that enable economic agents to achieve higher rates of productivity in open markets. Two major theories provide, to a large extent, the needed insights: the economic geography theory and the international business theory. The economic geography studies scrutinized in this study from Marshall to Porter, aim to explain the drivers of the concentration of specific industries and activities in specific locations. These activity agglomerations may be due to the creation of new enterprises, the expansion of existing firms, and the attraction of firms located elsewhere. Regarding this last possibility, the international business (IB) theories focus on the comparative advantages of locations as far as multinational enterprises (MNEs) strategies are concerned. According to international business theory, the comparative advantages of a location serves firms not only by exploiting their ownership advantages (mostly as far as market seeking, resource seeking and efficiency seeking investments are concerned) but also by augmenting and/or creating new ownership advantages (strategic asset seeking investments). The impact of a location on the competitiveness of firms is considered from both sides: the MNE’s home country and the MNE’s host country.

Keywords: competitiveness, economic geography, international business, attractiveness of businesses

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Authors: Stefano Pelaggi

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The research is focused on youth labor and mobility in Berlin. Mobility has become a common denominator in our daily lives but it does not primarily move according to monetary incentives. Labor, knowledge and leisure overlap on this point as cities are trying to attract people who could participate in production of the innovations while the new migrants are experiencing the lifestyle of the host cities. The research will present the project of empirical study focused on Italian workers in the digital industry in Berlin, trying to underline the connection between pleasure, leisure with the choice of life abroad. Berlin has become the epicenter of the European Internet start-up scene, but people suitable to work for digital industries are not moving in Berlin to make a career, most of them are attracted to the city for different reasons. This point makes a clear exception to traditional migration flows, which are always originated from a specific search of employment opportunities or strong ties, usually families, in a place that could guarantee success in finding a job. Even the skilled migration has always been originated from a specific need, finding the right path for a successful professional life. In a society where the lack of free time in our calendar seems to be something to be ashamed, the actors of youth mobility incorporate some categories of experiential tourism within their own life path. Professional aspirations, lifestyle choices of the protagonists of youth mobility are geared towards meeting the desires and aspirations that define leisure. While most of creative work places, in particular digital industries, uses the category of fun as a primary element of corporate policy, virtually extending the time to work for the whole day; more and more people around the world are deciding their path in life, career choices on the basis of indicators linked to the realization of the self, which may include factors like a warm climate, cultural environment. All indicators that are usually eradicated from the hegemonic approach to labor. The interpretative framework commonly used seems to be mostly focused on a dualism between Florida's theories and those who highlight the absence of conflict in his studies. While the flexibility of the new creative industries is minimizing leisure, incorporating elements of leisure itself in work activities, more people choose their own path of life by placing great importance to basic needs, through a gaze on pleasure that is only partially driven by consumption. The multi localism is the co-existence of different identities and cultures that do not conflict because they reject the bind on territory. Local loses its strength of opposition to global, with an attenuation of the whole concept of citizenship, territory and even integration. A similar perspective could be useful to search a new approach to all the studies dedicated to the gentrification process, while studying the new migrations flow.

Keywords: brain drain, digital industry, leisure and gentrification, multi localism

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Authors: Wessam H. Abd-Elsalam, Mona M. Saber, Samar M. Abouelatta

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Non-steroidal anti-inflammatory drugs (NSAIDs) are considered a double edged sword in inflammatory bowel diseases (IBDs). Some studies reported their advantageous effect in decreasing inflammation, and other studies reported that their use is associated with colitis aggravation. This study aimed to use specifically formulated trehalose-based nano-carriers that targets the colon in an attempt to alleviate inflammation caused by NSAIDs. L-α-phosphatidylcholine (PL), trehalose, and transcutol were used to prepare the trehalosomes (THs), which were also loaded with Tenoxicam(TXM) as a model NSAID. To optimize the formulation variables, a full 23 factorial design, using Design-Expert® software, was performed. The optimized formulation composed of trehalose: PL at a weight ratio of 1:1, 377.72 mg transcutol, and sonicated for 4 min, possessed a spherical shape with a size of 268.61 nm and EE% of 97.83% and released 70.22% of its drug content over 24 h. The superior protective action of TXM loaded THs compared to TXM suspension and drug-free THs was shown by the inhibition of the inflammatory biomarkers, namely; IL-1ß, IL-6, and TNF-alpha levels, as well as oxidative stress markers, measured as GSH and MDA. Improved histopathology of the colonic tissue in male New Zealand rabbits also confirmed the superiority of the TXM loaded THs compared to the unformulated drug or the drug free nano-carriers. Our findings highlight the prosperous role of THs in colon targeting and its anti-inflammatory characteristics in guarding against possible NSAIDs-driven exacerbation of colitis.

Keywords: inflammatory bowel disease, trehalose, trehalosomes, colon targeting

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Authors: Alex Hughes

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Patients with advanced heart failure develop specific palliative care needs due to the progressive symptom burden and unpredictable disease trajectory. NICE guidance advises that palliative care should be provided to patients with both cancer and non-cancer conditions as and when required. However, there is some way to go before this guidance is consistently and effectively implemented nationwide in conditions such as heart failure. The Ambitions for Palliative and End of Life Care: A national framework for local action in England provides a set of foundations and ambitions which outline a vision for what high-quality palliative and end-of-life care look like in England. This poster aims to critically consider how to improve access to palliative care for heart failure patients in England by analysing the foundations taken from this framework to generate specific recommendations using Soft Systems Methodology (SSM). The eight foundations analysed are: ‘Personalised care planning’, ‘Shared records’, ‘Evidence and information’, ‘Involving, supporting and caring for those important to the dying Person’, ‘Education and training’, ‘24/7 access’, ‘Co-design’ and ‘Leadership.’ A number of specific recommendations have been generated which highlight a need to close the evidence-policy gap and implement policy with sufficient evidence. These recommendations, alongside the creation of an evidence-based national strategy for palliative care and heart failure, should improve access to palliative care for heart failure patients in England. Once implemented, it will be necessary to evaluate the effect of these proposals to understand if access to palliative care for heart failure patients actually improves.

Keywords: access, health systems, heart failure, palliative care

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Authors: Surena V.

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Background and Aim: Periodontal diseases are among the most common infectious diseases all around the world, even in developed countries. Considering the increased rate of microbial resistance to antibiotics and the chemical side effects of antibiotics and antiseptics used for the treatment of periodontal disease, there is a need for an alternative antimicrobial agent with fewer complications. Medicinal herbs have recently become popular as antimicrobial and preventive agents. This study aimed to assess the antibacterial effects of hydroalcoholic extracts of Lawsonia inermis, Malva sylvestris and Boswellia serrata on Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans). Materials and Methods: Hydroalcoholic extracts of the three medicinal plants were obtained by the maceration technique and A. actinomycetemcomitans was cultured. The antimicrobial efficacy of the three medicinal plants was compared with that of 0.2% chlorhexidine (CHX) according to the CLSI protocol using agar disc diffusion and broth microdilution techniques. All tests were repeated three times. Results: Hydroalcoholic extracts of all three plants had antimicrobial activity against A. actinomycetemcomitans. The minimum inhibitory concentration (MIC) of Lawsonia inermis, Malva sylvestris, and Boswellia serrata was 78.1, 156.2, and 1666 µg/mL with no significant difference between them. The MIC of CHX was 3.33 µg/mL, which was significantly higher than that of Boswellia serrata extract. Conclusion: Given that, further in vivo studies confirm other properties of these extracts and their safety in terms of cytotoxicity and mutagenicity, hydroalcoholic extracts of Lawsonia inermis and Malva sylvestris may be used in mouthwashes or local delivery systems to affect periodontal biofilm.

Keywords: actinobacilus actinomycetem commitans, lawsonia inermis, malva sylvestris, boswellia serrata

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Authors: Rajvir Kaur, Jeewani Anupama Ginige

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With recent trends in Big Data and advancements in Information and Communication Technologies, the healthcare industry is at the stage of its transition from clinician oriented to technology oriented. Many people around the world die of cancer because the diagnosis of disease was not done at an early stage. Nowadays, the computational methods in the form of Machine Learning (ML) are used to develop automated decision support systems that can diagnose cancer with high confidence in a timely manner. This paper aims to carry out the comparative evaluation of a selected set of ML classifiers on two existing datasets: breast cancer and cervical cancer. The ML classifiers compared in this study are Decision Tree (DT), Support Vector Machine (SVM), k-Nearest Neighbor (k-NN), Logistic Regression, Ensemble (Bagged Tree) and Artificial Neural Networks (ANN). The evaluation is carried out based on standard evaluation metrics Precision (P), Recall (R), F1-score and Accuracy. The experimental results based on the evaluation metrics show that ANN showed the highest-level accuracy (99.4%) when tested with breast cancer dataset. On the other hand, when these ML classifiers are tested with the cervical cancer dataset, Ensemble (Bagged Tree) technique gave better accuracy (93.1%) in comparison to other classifiers.

Keywords: artificial neural networks, breast cancer, classifiers, cervical cancer, f-score, machine learning, precision, recall

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Authors: Mario C. Nierras

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This study aimed to determine the extension services’ needs of small farmers in Biliran province, Philippines. It also sought to find out other issues/concerns of the small farmers. Extension services’ needs of small farmers were gathered through personal interviewing and observational analysis of randomly-selected small farmers in Biliran, Philippines. Biliran small farmers extension services’ needs include: raising fruits, raising legumes, raising vegetables, raising swine, raising cattle, and raising chicken (as priority broad skills). For the specific skills, diagnosing symptoms on fertilizer deficiencies, controlling plant pests and diseases, diagnosing signs on specific pest and disease damage, controlling animal pests and diseases, and doing artificial insemination were the priority skills. They considered an on-farm trial of new technology as most needed to be coupled with industry and quality-orientedness, as positive behaviors needed in farming success. The farmers still adhere to the so-called wait-and-see attitude, thus they are more convinced to follow a particular technology if they see a concrete result of the introduced changes. Technical needs prioritization of Biliran small farmers showed that they have a real need for crop and animal production skills to include the other issues/concerns. Extension service program planning for small farmers should be patterned after their technical needs giving due attention to some issues/concerns so that extension work could deliver the right skills for the right needs of the farmers.

Keywords: extension, extension service, extension service needs, extension service program, farmers, small farmers, marginal farmers

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Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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