Search results for: Isolation

Authors: Haftay Abraha Tadesse, Atsebaha Gebrekidan Kahsay, Mahumd Abdulkader

Abstract:

Background: Salmonella species and Escherichia coli are important foodborne pathogens affecting humans and animals. They are among the most important causes of infection that are associated with the consumption of contaminated food. This study was aimed to determine the prevalence, antimicrobial susceptibility patterns and associated risk factors for Salmonella species and E. coli in raw meat from butchery houses of Mekelle, Northern Ethiopia. Methodology: A cross-sectional study was conducted from January to September 2019. Socio-demographic data and risk factors were collected using a predesigned questionnaire. Meat samples were collected aseptically from the butchery houses and transported using icebox to Mekelle University, College of Veterinary Sciences for the isolation and identification of Salmonella species and E. coli, Antimicrobial susceptibility patterns were determined using Kirby disc diffusion method. Data obtained were cleaned and entered into Statistical Package for the Social Sciences version 22 and logistic regression models with odds ratio were calculated. P-value < 0.05 was considered as statistically significant. Results: A total of 153 out of 384 (39.8%) of the meat specimens were found to be contaminated. The contamination of Salmonella species and E. coli were 15.6% (n=60) and 20.8%) (n=80), respectively. Mixed contamination (Salmonella species and E. coli) was observed in 13 (3.4 %) of the analyzed. Poor washing hands regularly (AOR = 8.37; 95% CI: 2.75-25.50) and not using gloves during meat handling (AOR=11. 28; 95% CI: (4.69 27.10) were associated with an overall bacterial contamination.About 95.5% of the tested isolates were sensitive to chloramphenicol and norfloxacin while the resistance of amoxyclav_amoxicillin and erythromycin were both isolated bacteria species. The overall multidrug resistance pattern for Salmonella and E. coli were 51.4% (n=19) and 31.8% (14), respectively. Conclusion: Of the 153 (153/384) contaminated raw meat, 60 (15.6%) and 80 (20.8%) were contaminated by Salmonella species and E. coli, respectively. Poor hand washing practice and not using glove during meat handling showed significant association with bacterial contamination. Multidrug-resistant showed in Salmonella species and E. coli were 19 (51.4%) and 14 (31.8%), respectively.

Keywords: antimicrobial susceptibility test, butchery houses, e. coli, salmonella species

Procedia PDF Downloads 14

Authors: Kumudini Apsara Munasinghe, Devin Gregory Lissau, Ryan Thomas Wade

Abstract:

Fresh fruits and vegetables are rich in vitamins, minerals, and fibers and help maintain a healthy weight over high-calorie food. Eating fruits and vegetables protects us from free radicals produced by metabolic reactions and safeguards us from cardiovascular disease and cancer. However, there has been an increased concern about foodborne diseases tied to contaminated farmers’ market produce. In addition, very little information is available about the contribution of eating raw fruits and vegetables to human exposure to antibiotic-resistant bacteria. This research aims to identify bacteria isolated from farmers’ market fruits and vegetables and understand their antibiotic resistance. Vegetables and fruits were collected from farmers’ markets around Frostburg and Cumberland areas in Maryland and transported to the microbiology lab at Frostburg State University for the isolation of bacteria. Bacteria were extracted from tomatoes, cucumber, strawberry, and lettuce using Tryptic soy broth overnight at 37°C, and Tryptic Soy agar was used for the streak plate technique to isolate bacteria. Pure cultures were used to identify bacteria using biochemical reactions after conducting Gram staining technique. The research used many biochemical reactions, including Mannitol Salt agar, MacConkey agar, and Eosin Methylene blue agar, for identification. Antibiotic sensitivity was tested for many different types of antibiotics, including amoxicillin, penicillin, tetracycline, ampicillin, and erythromycin. Most prevalent bacteria in the isolates were Staphylococcus, Bacillus, Micrococcus, Enterococcus, Enterobacter, Citrobacter, and other bacteria from the family Enterobacteriaceae. The data obtained from this research will be useful to educate and train farmers and individuals involved in post-harvest processes such as transportation and selling in farmers’ markets. Further results for bacterial antibiotic resistance will be obtained, and unculturable bacteria will be identified by next-generation DNA sequencing.

Keywords: antibiotic resistance, farmers markets, fruits, bacteria, vegetables

Procedia PDF Downloads 41

Authors: Małgorzata Tomczyńska, Joanna Saluk-Bijak

Abstract:

Background: Graves’ disease is a frequent organ-specific autoimmune thyroid disease, which characterized by the presence of different kind autoantibodies, that, in most cases, act as agonists of the thyrotropin receptor, leading to hyperthyroidism. Role of platelets and lymphocytes can be modulated in the pathophysiology of thyroid autoimmune diseases. Interference in the physiology of platelets can lead to enhanced activity of these cells. Activated platelets can bind to circulating lymphocytes and to affect lymphocyte adhesion. Platelets and lymphocytes can regulate mutual functions. Therefore, the activation of T lymphocytes, as well as blood platelets, is associated with the development of inflammation and oxidative stress within the target tissue. The present study was performed to investigate a platelet-lymphocyte relation by assessing the degree of their mutual aggregation in whole blood of patients with Graves’ disease. Also, the purpose of this study was to examine the impact of platelet interaction on lymphocyte migration capacity. Methods: 30 patients with Graves’ disease were recruited in the study. The matched 30 healthy subjects were served as the control group. Immunophenotyping of lymphocytes was carried out by flow cytometry method. A CytoSelect™ Cell Migration Assay Kit was used to evaluate lymphocyte migration and adhesion to blood platelets. Visual assessment of lymphocyte-platelet aggregate morphology was done using confocal microscope after magnetic cell isolation by Miltenyi Biotec. Results: The migration and functional responses of lymphocytes to blood platelets were greater in the group of Graves’ disease patients compared with healthy controls. The group of Graves’ disease patients exhibited a reduced T lymphocyte and a higher B cell count compared with controls. Based on microscopic analysis, more platelet-lymphocyte aggregates were found in patients than in control. Conclusions: Studies have shown that in Graves' disease, lymphocytes show increased platelet affinity, more strongly migrating toward them, and forming mutual cellular conglomerates. This may be due to the increased activation of blood platelets in this disease.

Keywords: blood platelets, cell migration, Graves’ disease, lymphocytes, lymphocyte-platelet aggregates

Procedia PDF Downloads 201

Authors: Armando A. Rodríguez, Emilio Salceda, Anoland Garateix, André J. Zaharenko, Steve Peigneur, Omar López, Tirso Pons, Michael Richardson, Maylín Díaz, Yasnay Hernández, Ludger Ständker, Jan Tytgat, Enrique Soto

Abstract:

Acid-sensing ion channels are cation (Na+) channels activated by a pH drop. These proteins belong to the ENaC/degenerin superfamily of sodium channels. ASICs are involved in sensory perception, synaptic plasticity, learning, memory formation, cell migration and proliferation, nociception, and neurodegenerative disorders, among other processes; therefore those molecules that specifically target these channels are of growing pharmacological and biomedical interest. Sea anemones produce a large variety of ion channels peptide toxins; however, those acting on ligand-gated ion channels, such as Glu-gated, Ach-gated ion channels, and acid-sensing ion channels (ASICs), remain barely explored. The peptide PhcrTx1 is the first compound characterized from the sea anemone Phymanthus crucifer, and it constitutes a novel ASIC inhibitor. This peptide was purified by chromatographic techniques and pharmacologically characterized on acid-sensing ion channels of mammalian neurons using patch-clamp techniques. PhcrTx1 inhibited ASIC currents with an IC50 of 100 nM. Edman degradation yielded a sequence of 32 amino acids residues, with a molecular mass of 3477 Da by MALDI-TOF. No similarity to known sea anemone peptides was found in protein databases. The computational analysis of Cys-pattern and secondary structure arrangement suggested that this is a structurally ICK (Inhibitor Cystine Knot)-type peptide, a scaffold that had not been found in sea anemones but in other venomous organisms. These results show that PhcrTx1 represents the first member of a new structural group of sea anemones toxins acting on ASICs. Also, this peptide constitutes a novel template for the development of drugs against pathologies related to ASICs function.

Keywords: animal toxin, inhibitor cystine knot, ion channel, sea anemone

Procedia PDF Downloads 274

Authors: Prathama Guha Chaudhuri, Arunima Datta, Ashis Mukhopadhyay

Abstract:

Background: Visualization (guided imagery) is a set of techniques which induce relaxation and help people create positive mental images in order to reduce stress.It is relatively inexpensive and can even be practised by bed bound people. Studies have shown visualization to be an effective tool to improve cancer patients’ anxiety, depression and quality of life. The common images used with cancer patients in the developed world are those involving the individual’s body and its strengths. Since breast cancer patients in India are more family oriented and often their main concerns are the stigma of having cancer and subsequent isolation of their families, including their children, we figured that positive images involving acceptance and integration within family and society would be more effective for them. Method: Data was collected from 119 breast cancer patients on chemotherapy willing to undergo psychotherapy, with no history of past psychiatric illness. Their baseline stress, anxiety, depression and quality of life were assessed using validated tools. The participants were then randomly divided into three groups: a) those who received visualization therapy with standard imageries involving the body and its strengths (sVT), b) those who received visualization therapy using indigenous family oriented imageries (mVT) and c) a control group who received supportive therapy. There were six sessions spread over two months for each group. The psychological outcome variables were measured post intervention. Appropriate statistical analyses were done. Results:Both forms of visualization therapy were more effective than supportive therapy alone in reducing patients’ depression, anxiety and quality of life.Modified VT proved to be significantly more effective in improving patients’ anxiety and quality of life. Conclusion: Visualization is a valuable therapeutic option for reduction of psychological distress and improving quality of life of breast cancer patients.In order to be more effective, the images used need to be modified according to the sociocultural background and individual needs of the patients.

Keywords: breast cancer, visualization therapy, quality of life, anxiety, depression

Procedia PDF Downloads 238

Authors: Howaida I. Abd-Alla, Omnia Kutkat, Yassmin Moatasim, Magda T. Ibrahim, Marwa A. Mostafa, Mohamed GabAllah, Mounir M. El-Safty

Abstract:

Artemisia judaica (known shih-balady), Azadirachta indica and Sophora tomentosa (known yellow necklace pod) are members of available medicinal plants well-known for their traditional medical use in Egypt which suggests that they probably harbor broad-spectrum antiviral, immunostimulatory and anti-inflammatory functions. Their ethyl acetate-dichloromethane (1:1, v/v) extracts were evaluated for the potential anti-Middle East respiratory syndrome-related coronavirus (anti-MERS-CoV) activity. Their cytotoxic activity was tested in Vero-E6 cells using 3-(4,-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method with minor modification. The plot of percentage cytotoxicity for each extract concentration has calculated the concentration which exhibited 50% cytotoxic concentration (TC50). A plaque reduction assay was employed using safe dose of extract to evaluate its effect on virus propagation. The highest inhibition percentage was recorded for the yellow necklace pod, followed by Shih-balady. The possible mode of action of virus inhibition was studied at three different levels viral replication, viral adsorption and virucidal activity. The necklace pod leaves have induced virucidal effects and direct effects on the replication of virus. Phytochemical investigation of the promising necklace pod led to the isolation and structure determination of nine compounds. The structure of each compound was determined by a variety of spectroscopic methods. Compounds 4-O-methyl sorbitol 1, 8-methoxy daidzin 6 and 6-methoxy apigenin-7-O-β-D-glucopyranoside 8 were isolated for the first time from the Sophora genus and the other six compounds were the first time that they were isolated from this species according to available works of literature. Generally, the highest anti-CoV 2 activity of S. tomentosa was associated with the crude ethanolic extract, indicating the possibility of synergy among the antiviral phytochemical constituents (1-9).

Keywords: coronavirus, MERS-CoV, mode of action, necklace pod, shih-balady

Procedia PDF Downloads 180

Authors: Gerhardt Boukes, Maryna Van De Venter, Sharlene Govender

Abstract:

Macrofungi have been used for the past two thousand years in Asian countries, and more recently in Western countries, for their medicinal properties. Biological activities include antimicrobial, antioxidant, anti-inflammatory, antidiabetic, anticancer and immunomodulatory to name a few. Several biologically active compounds have been identified and isolated. Macrofungal research in Africa is poorly documented and to the best of our knowledge non-existent. South Africa has a rich macrofungal biodiversity, which includes endemic and exotic macrofungal species. Ethanolic extracts of 13 macrofungal species, including mushrooms, bracket fungi and puffballs, were prepared and screened for cytotoxicity against a panel of seven cell lines, including A549 (human lung adenocarcinoma), HeLa (human cervical adenocarcinoma), HT-29 (human colorectal adenocarcinoma), MCF7 (human breast adenocarcinoma), MIA PaCa-2 (human pancreatic ductal adenocarcinoma), PC-3 (human prostate adenocarcinoma) and Vero (African green monkey kidney epithelial) cells using MTT. Cell lines were chosen according to the most prevalent cancer types affecting males and females in South Africa and globally, and the mutations they contain. Preliminary results have shown that three of the macrofungal genera, i.e. Fomitopsis, Gymnopilus and Pycnoporus, have shown cytotoxic activity, ranging between IC50 ~20 and 200 µg/mL. The molecular mechanism of action contributing to cell death investigated and being investigated include apoptosis (i.e. DNA cell cycle arrest, caspase-3 activation and mitochondrial membrane potential), autophagy (i.e. acridine orange and LC3B staining) and ER stress (i.e. thioflavin T staining and caspase-12) in the presence of melphalan, chloroquine and thapsigargin/tuncamycin as positive controls, respectively. The genus, Pycnoporus, has shown the best cytotoxicity of the three macrofungal genera. Future work will focus on the identification and isolation of novel active compounds and elucidating the mechanism/s of action.

Keywords: cancer, cytotoxicity, macrofungi, mechanism/s of action

Procedia PDF Downloads 219

Authors: Tamara Tallman

Abstract:

Collaboration is a powerful tool for professional development and central for creating opportunities for teachers to reflect on their practice. However, school districts continue to have difficulty both implementing and sustaining collaboration. The purpose of this research was to investigate the experience of the teacher in a creative, instructional collaboration. The teachers in this study found that teacher-initiated collaboration offered them trust and they were more open with their partners. An interpretative phenomenological analysis was used for this study as it told the story of the teacher’s experience. Interpretative Phenomenological Analysis was chosen for this study to capture the complex and contextual nature of the teacher experience from a creative, instructional collaborative experience. This study sought to answer the question of how teachers in a private, faith-based school experience collaboration. In particular, the researcher engaged the study’s participants in interviews where they shared their unique perspectives on their experiences in relation to this phenomenon. Through the use of interpretative phenomenological analysis, the researcher interpreted the experiences of each participant in an attempt to gain deeper insight into how teachers made sense of their understanding of collaboration. In addition to the researcher’s interpreting the meaning of this construct for each research participant, this study gave a voice to the individual experiences and positionality of each participant at the research site. Moreover, the key findings presented in this study shed light on how teachers within this particular context participated in and made sense of their experience of creating an instructional collaborative. The research presented the findings that speak to the meaning that each research participant experienced in their relation to participating in building a collaborative culture and its effect on professional and personal growth. The researcher provided recommendations for future practice and research possibilities. The research findings demonstrated the unique experiences of each participant as well as a connection to the literature within the field of teacher professional development. The results also supported the claim that teacher collaboration can facilitate school reform. Participating teachers felt less isolation and developed more teacher knowledge.

Keywords: collaboration, personal grwoth, professional development, teachers

Procedia PDF Downloads 88

Authors: Bharati Mehta, Manish Parakh, Bharti Bhandari, Sneha Ambwani

Abstract:

Speech and language delay (SLD) is seen commonly as a co-morbidity in children having severe resistant focal and generalized, syndromic and symptomatic epilepsies. It is however not clear whether epilepsy contributes to or is a mere association in the pathogenesis of SLD. Also, it is acknowledged that Auditory Brainstem Evoked Responses (ABER), besides used for evaluating hearing threshold, also aid in prognostication of neurological disorders and abnormalities in the hearing pathway in the brainstem. There is no circumscribed or surrogate neurophysiologic laboratory marker to adjudge the extent of SLD. The current study was designed to evaluate the abnormalities in Electroencephalography (EEG) and ABER in children with SLD who do not have an overt hearing deficit or autism. 94 children of age group 2-8 years with predominant SLD and without any gross motor developmental delay, head injury, gross hearing disorder, cleft lip/palate and autism were selected. Standard video Electroencephalography using the 10:20 international system and ABER after click stimulus with intensities 110 db until 40 db was performed in all children. EEG was abnormal in 47.9% (n= 45; 36 boys and 9 girls) children. In the children with abnormal EEG, 64.5% (n=29) had an abnormal background, 57.8% (n=27) had presence of generalized interictal epileptiform discharges (IEDs), 20% (n=9) had focal epileptiform discharges exclusively from left side and 33.3% (n=15) had multifocal IEDs occurring both in isolation or associated with generalised abnormalities. In ABER, surprisingly, the peak latencies for waves I, III & V, inter-peak latencies I-III & I-V, III-V and wave amplitude ratio V/I, were found within normal limits in both ears of all the children. Thus in the current study it is certain that presence of generalized IEDs in EEG are seen in higher frequency with SLD and focal IEDs are seen exclusively in left hemisphere in these children. It may be possible that even with generalized EEG abnormalities present in these children, left hemispheric abnormalities as a part of this generalized dysfunction may be responsible for the speech and language dysfunction. The current study also emphasizes that ABER may not be routinely recommended as diagnostic or prognostic tool in children with SLD without frank hearing deficit or autism, thus reducing the burden on electro physiologists, laboratories and saving time and financial resources.

Keywords: ABER, EEG, speech, language delay

Procedia PDF Downloads 494

Authors: Fang Liu, JunShuai Xue, JiaJia Yao, XueYan Yang, ZuMao Li, GuanLin Wu, HePeng Zhang, ZhiPeng Sun

Abstract:

III-Nitride resonant tunneling diode (RTD) is one of the most promising candidates for multiple-valued logic (MVL) elements. Here, we report a monolithic integration of GaN resonant tunneling diodes to realize multiple negative differential resistance (NDR) regions for MVL application. GaN RTDs, composed of a 2 nm quantum well embedded in two 1 nm quantum barriers, are grown by plasma-assisted molecular beam epitaxy on free-standing c-plane GaN substrates. Negative differential resistance characteristic with a peak current density of 178 kA/cm² in conjunction with a peak-to-valley current ratio (PVCR) of 2.07 is observed. Statistical properties exhibit high consistency showing a peak current density standard deviation of almost 1%, laying the foundation for the monolithic integration. After complete electrical isolation, two diodes of the designed same area are connected in series. By solving the Poisson equation and Schrodinger equation in one dimension, the energy band structure is calculated to explain the transport mechanism of the differential negative resistance phenomenon. Resonant tunneling events in a sequence of the series-connected RTD pair (SCRTD) form multiple NDR regions with nearly equal peak current, obtaining three stable operating states corresponding to ternary logic. A frequency multiplier circuit achieved using this integration is demonstrated, attesting to the robustness of this multiple peaks feature. This article presents a monolithic integration of SCRTD with multiple NDR regions driven by the resonant tunneling mechanism, which can be applied to a multiple-valued logic field, promising a fast operation speed and a great reduction of circuit complexity and demonstrating a new solution for nitride devices to break through the limitations of binary logic.

Keywords: GaN resonant tunneling diode, multiple-valued logic system, frequency multiplier, negative differential resistance, peak-to-valley current ratio

Procedia PDF Downloads 54

Authors: Henry Kelly, Helena Shaw

Abstract:

This paper will seek how innovative methods from Health Economics and, to a lesser extent, wellbeing analysis can be applied in the Cost-Benefit Analysis (CBA) of transport infrastructure and policy interventions. The context for this will focus on the framework articulated by the UK Treasury (finance department) and the English Department for Transport. Both have well-established methods for undertaking CBA, but there is increased policy interest, particularly at a regional level of exploring broader strategic goals beyond those traditionally associated with transport user benefits, productivity gains, and labour market access. Links to different CBA approaches internationally, such as New Zealand, France, and Wales will be referenced. By exploring a complementary method of accessing the impacts of policies through the quantification of health impacts is a fruitful line to explore. In a previous piece of work, 14 impact pathways were identified, mapping the relationship between transport and health. These are wide-ranging, from improved employment prospects, the stress of unreliable journey times, and air quality to isolation and loneliness. Importantly, we will consider these different measures of health from an intersectional point of view to ensure that the basis that remains in the health industry does not get translated across to this work. The objective is to explore how a CBA based on these pathways may, through quantifying forecast impacts in terms of Quality-Adjusted Life Years may, produce different findings than a standard approach. Of particular interest is how a health-based approach may have different distributional impacts on socio-economic groups and may favour distinct types of interventions. Consideration will be given to the degree this approach may double-count impacts or if it is possible to identify additional benefits to the established CBA approach. The investigation will explore a range of schemes, from a high-speed rail link, highway improvements, rural mobility hubs, and coach services to cycle lanes. The conclusions should aid the progression of methods concerning the assessment of publicly funded infrastructure projects.

Keywords: cost-benefit analysis, health, QALYs transport

Procedia PDF Downloads 59

Authors: Fae Princess Bermudez

Abstract:

Background: One of the least significant leads on a 12-lead electrocardiogram is the augmented right lead (aVR), as it is not as specific compared to the other leads. In this case series, the value of lead aVR, which is more often than not ignored, is highlighted. Three cases of aVR ST segment elevation on 12-lead electrocardiogram are described, with the end outcome of demise of all three patients. The importance of immediate revascularization is described to improve prognosis in this group of patients. Objectives: This case series aims to primarily present under-reported cases of isolated aVR ST-segrment elevation myocardial infarction (STEMI), their course and outcome. More specific aims are to identify the criteria in determination of isolated aVR STEMI, know its clinical significance, and determine appropriate management for patients with this ECG finding. Method: A short review of previous studies, case reports, articles and guidelines from 2011-2016 was done. The author reviewed available literature, sorted out those that proved to be significant for the presented cases, and described them in conjunction with the aforementioned cases. Findings: Based on the limited information on these rare or under-reported cases, it was found that isolated aVR STEMI had a poorer prognosis that led to significant mortality and morbidity of patients. The significance of aVR ST-elevation was that of an occlusion of the left coronary artery or a severe three-vessel disease in the presence of an Acute Coronary Syndrome. Guidelines from American Heart Association/American College of Cardiology Foundation in 2013 already recognized ST-elevation of lead aVR in isolation as a STEMI; hence, recommended that patients with this particular ECG finding should undergo reperfusion strategies to improve prognosis. Conclusion: The indispensability of isolated aVR ST-segment elevation on ECG should alert physicians, especially Emergency physicians, to the high probability of Acute Coronary Syndrome with a very poor prognosis. If this group of patients is not promptly managed, demise may ensue, with cardiogenic shock as the most probable cause. With this electrocardiogram finding, physicians must be quick to make clinical decisions to increase chances of survival of this group of patients.

Keywords: AVR ST-elevation, diffuse ST-segment depression, left coronary artery infarction, myocardial infarction

Procedia PDF Downloads 187

Authors: Denis Mustafov, Emmanouil Karteris, Maria Braoudaki

Abstract:

Glioblastoma multiform (GBM) is a heterogenous primary brain tumour that kills most affected patients. To the authors best knowledge, despite all research efforts there is no early diagnostic biomarker for GBM. MicroRNAs (miRNAs) are short non-coding RNA molecules which are deregulated in many cancers. The aim of this research was to determine miRNAs with a diagnostic impact and to potentially identify promising therapeutic targets for glioblastoma multiform. In silico analysis was performed to identify deregulated miRNAs with diagnostic relevance for glioblastoma. The expression profiles of the chosen miRNAs were then validated in vitro in the human glioblastoma cell lines A172 and U-87MG. Briefly, RNA extraction was carried out using the Trizol method, whilst miRNA extraction was performed using the mirVANA miRNA isolation kit. Quantitative Real-Time Polymerase Chain Reaction was performed to verify their expression. The presence of five target proteins within the A172 cell line was evaluated by Western blotting. The expression of the CORO1C protein within 32 GBM cases was examined via immunohistochemistry. The miRNAs identified in silico included miR-21-5p, miR-34a and miR-128a. These miRNAs were shown to target deregulated GBM genes, such as CDK6, E2F3, BMI1, JAG1, and CORO1C. miR-34a and miR-128a showed low expression profiles in comparison to a control miR-RNU-44 in both GBM cell lines suggesting tumour suppressor properties. Opposing, miR-21-5p demonstrated greater expression indicating that it could potentially function as an oncomiR. Western blotting revealed expression of all five proteins within the A172 cell line. In silico analysis also suggested that CORO1C is a target of miR-128a and miR-34a. Immunohistochemistry demonstrated that 75% of the GBM cases showed moderate to high expression of CORO1C protein. Greater understanding of the deregulated expression of miR-128a and the upregulation of CORO1C in GBM could potentially lead to the identification of a promising diagnostic biomarker signature for glioblastomas.

Keywords: non-coding RNAs, gene expression, brain tumours, immunohistochemistry

Procedia PDF Downloads 59

Authors: Elida Cena

Abstract:

Return migration is receiving increased attention as the phenomenon challenges assumptions of natural ‘homecomings’. This talk outlines preliminary findings from an ongoing PhD study which explores return migration of Albanian migrants (aged 30-50 years) and their children (aged 7-18 years). Participants (n=51) were purposively recruited from two Albanian cities with divergent social and economic conditions, and the majority had returned from Greece following the recent economic downturn in that country. Qualitative data were collected through in-depth interviews with respondents aged 13 years and above, and were augmented with focus groups and family case studies. Data collection for case studies was aided by photo elicitation, interviews and participatory techniques (drawing) were employed for children aged 7-12 years. Through a multidisciplinary perspective, findings will uncover experiences of migrants and children upon return, the quest to identify with the originating country and create a sense of belongingness. Narrative analysis reveals that the abrupt return was associated with ambivalent feelings and disillusionment about their (re)settlement for both younger and older participants. Faced with unexpected realities and lack of opportunities, particularly for the children of migrants, Albania is viewed as a ‘transit country’, a temporary solution to escape the crisis in the destination country and move to a more developed western country. Adult return migrants articulate lack of employment and insecurity for the future. Apart from school difficulties, children experience isolation and social exclusion, marked by stigmatized labelling from other peers which exacerbates their belonging. Such mobilities have had deeper effects in complicating family relationships as influenced by many disintegration factors. Feelings of alienation and being emigrant for the second time were common in participants' accounts. Findings concerning the difficulties of individuals (re)connecting with their ethnic background and the impact on their identities are discussed in relation to the literature on return migration and identification.

Keywords: return migration, belonging, identity, disintegration, integration

Procedia PDF Downloads 345

Authors: Anna Siegler

Abstract:

Bullying among peers is not uncommon; however, adults can notice only a fragment of the cases of harassment during everyday life. The systemic approaches of bullying investigation put the whole school community in the focus of attention and propose that the solution should emerge from the culture of the school. Bystanders are essential in the prevention and intervention processes as an active agent rather than passive. For combating exclusion, stigmatization and harassment, it is important that the bystanders have to realize they have the power to take action. To prevent the escalation of violence, victims must believe that students and teachers will help them and their environment is able to provide safety. The study based on scientific narrative psychological approach, and focuses on the examination of the different perspectives of students, how peers are mentalizing with each other in case of bullying. The data collection contained responses of students (N = 138) from three schools in Hungary, and from three different area of the country (Budapest, Martfű and Barcs). The test battery include Bullying Prevalence Questionnaire, Interpersonal Reactivity Index and an instruction to get narratives about bullying, which effectiveness was tested during a pilot test. The obtained results are in line with the findings of previous bullying research: the victims are mentalizing less with their peers and experience greater personal distress when they are in identity threatening situations, thus focusing on their own difficulties rather than social signals. This isolation is an adaptive response in short-term although it seems to lead to a deficit in social skills later in life and makes it difficult for students to become socially integrated to society. In addition the results also show that students use more mental state attribution when they report verbal bullying than in case of physical abuse. Those who witness physical harassment also witness concrete answers to the problem from teachers, in contrast verbal abuse often stays without consequences. According to the results students mentalizing more in these stories because they have less normative explanation to what happened. To expanding bullying literature, this research helps to find ways to reduce school violence through community development.

Keywords: bullying, mentalization, narrative, school culture

Procedia PDF Downloads 138

Authors: Mohamed Trigui, Fatma Masmoudi, Imen Zouari

Abstract:

Massive utilizations of chemical fertilizer and chemical pesticides in agriculture sector to improve the farming productivity have created increasing environmental damages. Then, agriculture must become sustainable, focusing on production systems that respect the environment and help to reduce climate change. Isolation and microbial identification of new bacterial strains from naturally saline habitats and compost extracts could be a prominent way in pest management and crop production under saline conditions. In this study, potential mechanisms involved in plant growth promotion and suppressive activity against fungal diseases of a compost extract produced from poultry manure/olive husk compost and halotolerant and halophilic bacterial strains under saline stress were investigated. On the basis of the antimicrobial tests, different strains isolated from Sfax solar saltern (Tunisia) and from compost extracts were selected and tested for their plant growth promoting traits, such as siderophores production, nitrogen fixation, phosphate solubilization and the production of extracellular hydrolytic enzymes (protease and lipase) under in-vitro conditions. Among 450 isolated bacterial strains, 16 isolates showed potent antifungal activity against the tested plant pathogenic fungi. Their identification based on 16S rRNA gene sequence revealed they belonged to different species. Some of these strains were also characterized for their plant growth promoting capacities. Obtained results showed the ability of four strains belonging to Bacillus genesis to ameliorate germination rate and root elongation compared to the untreated positive controls. Combinatorial capacity of halotolerant bacteria with antimicrobial activity and plant growth promoting traits could be promising sources of interesting bioactive substances under saline stress.

Keywords: abiotic stress, biofertilizer, biotic stress, compost extract, halobacteria, plant growth promoting (PGP), soil fertility

Procedia PDF Downloads 60

Authors: Shagufta Tazin Shathy, Mohammad Imam Hasan Reza

Abstract:

Fragmentation due to urbanization and agricultural expansion has become the main reason for destruction of forest area and loss of biodiversity particularly in the developing world. At present, the world is experiencing the largest wave of urban growth in human history, and it is estimated that this influx will be mainly taking place in developing world. Therefore, study on urban fragmentation is vital for a sustainable urban development. Landscape fragmentation is one of the most important conservation issues in the last few decades. Habitat fragmentation due to landscape alteration has caused habitat isolation, destruction in ecosystem pattern and processes. Thus, this research analyses the spatial and temporal extent of urban fragmentation using landscape indices in the Kuala Lumpur (KL) – the capital and most populous city in Malaysia. The objective of this study is to examine the urban fragmentation index in KL city. Fragmentation metrics used in the study are: a) Urban landscape ratio (the ratio of urban landscape area and build up area), b) Infill (development that occurred within urbanized open space), and c) Extension (development of exterior open space). After analyzing all three metrics, these are calculated for the combined urban fragmentation index (UFI). In this combined index, all three metrics are given an equal weight. Land cover/ land use maps of the year 1996 and 2005 have been developed from the Landsat TM 30 m resolution satellite image. The year 1996 is taken as a reference year to analyze the changes. The UFI calculated for the year of 1996 and2005 found that the KL city has undergone rapid landscape changes destructing forest ecosystem adversely. Increasing UFI for the year of 1996 compared to 2005 indicates that the developmental activities have been occupying open spaces and fragmenting natural lands and forest. This index can be implemented in other unplanned and rapidly urbanizing Asian cities for example Dhaka and Delhi to calculate the urban fragmentation rate. The findings from the study will help the stakeholders and urban planners for a sustainable urban management planning in this region.

Keywords: GIS, index, sustainable urban management, urbanization

Procedia PDF Downloads 347

Authors: Marija Vitanova, Aleksandra Bogdanovic, Kemal Edip, Viktor Hristovski, Vlado Micov

Abstract:

Bridges represent critical structures in lifeline systems. They provide reliable modes of transportation, so their failure can seriously obstruct relief and rehabilitation work. Earthquake ground motions can cause significant damages in bridges, so during the strong earthquakes, they can easily collapse. The base isolation technique has been quite effective in seismic response mitigation of the bridges in reducing the piers base shear. The effect of soil structure interaction on the dynamic responses of seismically isolated three span girder bridge with viscous dampers is investigated. Viscous dampers are installed in the mid span of the bridge to control bearing displacement. The soil surrounding the foundation of piers has been analyzed by applying different soil densities in order to consider the soil stiffness. The soil medium has been assumed as a four layered infill as dense and loose medium. The boundaries in the soil medium are considered as infinite elements in order to absorb the radiating waves. The formulation of infinite elements is the same as for the finite elements in addition to the mapping of the domain. Based on the iso-parametric concept, the infinite element in global coordinate is mapped onto an element in local coordinate system. In the formulation of the infinite element, only the positive direction extends to infinity thus allowing the waves to propagate outside of the soil medium. Dynamic analyses for two levels of earthquake intensity are performed in time domain using direct integration method. In order to specify the effects of the SSI, the responses of the isolated and controlled isolated bridges are compared. It is observed that the soil surrounding the piers has significant effects on the bearing displacement of the isolated RC bridges. In addition, it is observed that the seismic responses of isolated RC bridge reduced significantly with the installation of the viscous dampers.

Keywords: viscous dampers, reinforced concrete girder bridges, seismic response, SSI

Procedia PDF Downloads 98

Authors: Belinda Vega, Claudio Alvarez, Héctor Flores, Marcia Oliva, Katherine Alveal, Teresa Toro, María José Tapia, Fanny Guzmán

Abstract:

With the increase in global temperatures and the decrease of oxygen (O2) concentration in the oceans, fish cultures are exposed to frequent fluctuations in dissolved O2 (DO) concentration that can cause chronic stress in the animals, altering the normal functioning of their immune system and making them vulnerable to infections, consequently increasing morbidity and mortality in the farms with economic losses. The mucosal organs (skin -and mucus-, gills, gut, and nasal mucosa) are the first line of defense of the fish against pathogens. Therefore, the objective of this study is to evaluate the effect of hypoxia on the antimicrobial activity of epidermal mucus from corvina drum (Cilus Gilberti), a native marine species with the potential for the diversification of aquaculture in Chile. To achieve this, the epidermal mucus of juveniles (~220g) kept under normoxia (7 mg/L DO) and hypoxia (2 mg/L DO) environmental conditions was collected after 6 weeks, as well as after 6 days of intraperitoneal inoculation with lipopolysaccharide from Vibrio anguillarum to induce an immune response in the fish. Total protein extracts of the mucus were used for bactericidal activity and lysozyme and peroxidase activity assays. Although the mucus from both experimental groups showed inhibitory effects on the bacterial growth of Vibrio anguillarum and Vibrio ordalli, this effect was more long-lasting in the normoxia group. We also observed a notable reduction in the presence of lysozyme in the mucus from fish exposed to hypoxia, with no differences in peroxidase content. Future proteomic studies of corvina mucus associated with the environmental conditions studied in this work will allow the isolation and identification of peptides with antimicrobial activity, those responsible for the results obtained. This will help establish strategies aimed at minimizing the impacts of hypoxia on the defense responses of corvina drum against potential pathogens. Funding: FONDECYT 3200440 and FONDECYT 1210056

Keywords: Cilus gilberti, mucus, antimicrobial activity, HYPOXIA

Procedia PDF Downloads 49

Authors: Nicholas Bayfield, Liam Bibo, Kaushelandra Rathore, Lucas Sanders, Mark Newman

Abstract:

INTRODUCTION: Surgical pulmonary embolectomy is an established therapy for acute pulmonary embolism causing right heart dysfunction and haemodynamic instability. Massive intraoperative airway haemorrhage is a rare complication of pulmonary embolectomy. We present our institutional experience with massive airway haemorrhage complicating pulmonary embolectomy and discuss optimal therapeutic strategies. METHODS: A retrospective review of emergent surgical pulmonary embolectomy patients was undertaken. Cases complicated by massive intra-operative airway haemorrhage were identified. Intra- and peri-operative management strategies were analysed and discussed. RESULTS: Of 76 patients undergoing emergent or salvage pulmonary embolectomy, three cases (3.9%) of massive intraoperative airway haemorrhage were identified. Haemorrhage always began on weaning from cardiopulmonary bypass. Successful management strategies involved intraoperative isolation of the side of bleeding, occluding the affected airway with an endobronchial blocker, institution of veno-arterial (VA) extracorporeal membrane oxygenation (ECMO) and reversal of anticoagulation. Running the ECMO without heparinisation allows coagulation to occur. Airway haemorrhage was controlled within 24 hours of operation in all patients, allowing re-institution of dual lung ventilation and decannulation from ECMO. One case in which positive end-expiratory airway pressure was trialled initially was complicated by air embolism. Although airway haemorrhage was controlled successfully in all cases, all patients died in-hospital for reasons unrelated to the airway haemorrhage. CONCLUSION: Massive intraoperative airway haemorrhage during pulmonary embolectomy is a rare complication with potentially catastrophic outcomes. Re-perfusion alveolar and capillary injury is the likely aetiology. With a systematic approach to management, airway haemorrhage can be well controlled intra-operatively and often resolves within 24 hours. Stopping blood flow to the pulmonary arteries and support of oxygenation by the institution of VA ECMO is important. This management has been successful in our 3 cases.

Keywords: pulmonary embolectomy, cardiopulmonary bypass, cardiac surgery, pulmonary embolism

Procedia PDF Downloads 158

Authors: Maryam Farzaneh, Seyyedeh Nafiseh Hassani, Hossein Baharvand

Abstract:

Objective: Chicken gonadal tissue has a two population such primordial germ cells (PGCs) and stromal cells (somatic cells). PGCs and embryonic germ cells (EGCs) that is a pluripotent type of PGCs in long-term culture are suitable sources for the production of chicken pluripotent stem cell lines, transgenic birds, vaccine and recombinant protein production. In general, the effect of growth factors such bFGF and mouse LIF on derivation of PGCs in vitro are important and in this study we could see the unique effect of small molecules such PD032 and SB43 as a chemical, in comparison to growth factors. Materials and Methods: After incubation of fertilized chicken egg up to 6 days and isolation of primary gonadal tissues and culture of mixed cells like PGCs and stromal cells. PGCs proliferate in the present of fetal calf serum (FCS) and small molecules and in another group bFGF, that these factors are important for PGCs culture and derivation. Somatic cells produce a multilayer feeder under the PGCs in primary culture and PGCs make a small cluster under these cells. Results: In present of small molecules and high volume of FCS (15%), the present of EGCs as a pluripotent stem cells were clear four weeks, that they had a positive immune-staining and periodic acid-Schiff staining (PAS), but in present of growth factors like bFGF without any chemicals, the present of PGCs were clear but after 7 until 10 days, there were disappear. Conclusion: Until now we have seen many researches about derivation and maintenance of chicken PGCs, in the hope of understanding the mechanisms that occur during germline development and production of a therapeutic product by transgenic birds. There are still many unknowns in this area and this project will try to have efficient conditions for identification of suitable culture medium for long-term culture of PGCs in vitro without serum and feeder cells.

Keywords: chicken gonadal primordial germ cells, pluripotent stem cells, growth factors, small molecules, transgenic birds

Procedia PDF Downloads 409

Authors: Cengiz Demir, Recep Keşli, Gülşah Aşık

Abstract:

Objective: This study was carried out with the purpose of defining by using the E test method and determining the antibiotic resistance profiles of Gram-negative anaerobic bacilli isolated from various clinical specimens obtained from patients with suspected anaerobic infections and referred to Medical Microbiology Laboratory of Afyon Kocatepe University, ANS Application and Research Hospital. Methods: Two hundred and seventy eight clinical specimens were examined for isolation of the anaerobic bacteria in Medical Microbiology Laboratory between the 1st November 2014 and 30th October 2015. Specimens were cultivated by using Scheadler agar that 5% defibrinated sheep blood added, and Scheadler broth. The isolated anaerobic Gram-negative bacilli were identified conventional methods and Vitek 2 (ANC ID Card, bioMerieux, France) cards. Antibiotic resistance rates against to penicillin G, clindamycin, cefoxitin, metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem were determined with E-test method for each isolate. Results: Of the isolated twenty-eight anaerobic gram negative bacilli fourteen were identified as the B. fragilis group, 9 were Prevotella group, and 5 were Fusobacterium group. The highest resistance rate was found against penicillin (78.5%) and resistance rates against clindamycin and cefoxitin were found as 17.8% and 21.4%, respectively. Against to the; metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem, no resistance was found. Conclusion: Since high rate resistance has been detected against to penicillin in the study penicillin should not be preferred in empirical treatment. Cefoxitin can be preferred in empirical treatment; however, carrying out the antibiotic sensitivity testing will be more proper and beneficial. No resistance was observed against carbapenem group antibiotics and metronidazole; so that reason, these antibiotics should be reserved for treatment of infectious caused by resistant strains in the future.

Keywords: anaerobic gram-negative bacilli, anaerobe, antibiotics and resistance profiles, e-test method

Procedia PDF Downloads 277

Authors: Bathmajaa Muralisankar

Abstract:

As much as railway station infrastructure designs and ticket-booking norms have been changed to facilitate use by differently abled persons, the railway train compartments themselves have not been made user-friendly for differently abled persons. Owing to safety concerns, dependency on others for their travel, and fear of isolation, differently abled people do not prefer travelling by train. Rather than including a dedicated compartment open only to the differently abled, including the latter with others in the normal compartment (with the proposed modifications discussed here) will make them feel secure and make for an enhanced travel experience for them. This approach also represents the most practical way to include a particular category of people in the mainstream society. Lowering the height of the compartment doors and providing a wider entrance with a ramp will provide easy entry for those using wheelchairs. As well, removing the first two alternate rows and the first two side seats will not only widen the passage and increase seating space but also improve wheelchair turning radius. This will help them travel without having to depend on others. Seating arrangements may be done to accommodate their family members near them instead of isolating the differently abled in a separate compartment. According to present ticket-booking regulations of the Indian Railways, three to four disabled persons may travel without their family or one to two along with their family, and the numbers may be added or reduced. To help visually challenged and hearing-impaired persons, in addition to the provision of special instruments, railings, and textured footpaths and flooring, the seat numbers above the seats may be set in metal or plastic as an outward projection so the visually impaired can touch and feel the numbers. Braille boards may be included at the entrance to the compartment along with seat numbers in the aforementioned projected manner. These seat numbers may be designed as buttons, which when pressed results in an announcement of the seat number in the applicable local language as well as English. Emergency buttons, rather than emergency chains, within the easy reach of disabled passengers will also help them.

Keywords: dependency, differently abled, inclusion, mainstream society

Procedia PDF Downloads 234

Authors: Abd El Rahman Mohamed

Abstract:

Canopic jars made by the ancient Egyptians from different materials were used to preserve the viscera during the mummification process. The canopic jar studied here dates back to the Late Period (712-332 BC). It is found in the Grand Egyptian Museum (GEM), Giza, Egypt. This jar was carved from limestone and covered with a monkey head lid with painted eyes and ears with red pigment and surrounded with black pigment. The jar contains bandages of textile containing mummy viscera with resin and black resin blocks. The canopic jars were made using the sculpting tools that were used by the ancient Egyptians, such as metal chisels (made of copper) and hammers and emptying the mass of the jar from the inside using a tool invented by the ancient Egyptians, which called the emptying drill. This study also aims to use analytical techniques to identify the components of the jar, its contents, pigments, and previous restoration materials and to understand its deterioration aspects. Visual assessment, isolation and identification of fungi, optical microscopy (OM), scanning electron microscopy (SEM), X-ray fluorescence spectroscopy (XRF), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) were used in our study. The jar showed different signs of deterioration, such as dust, dirt, stains, scratches, classifications, missing parts, and breaks; previous conservation materials include using iron wire, completion mortar and an adhesive for assembly. The results revealed that the jar was carved from Dolomite Limestone, red Hematite pigment, Mastic resin, and Linen textile bandages. The previous adhesive was Animal Glue and used Gypsum for the previous completion. The most dominant Microbial infection on the jar was found in the fungi of (Penicillium waksmanii), (Nigrospora sphaerica), (Actinomycetes sp) and (Spore-Forming Gram-Positive Bacilli). Conservation procedures have been applied with high accuracy to conserve the jar, including mechanical and chemical cleaning, re-assembling, completion and consolidation.

Keywords: Canopic jar, Consolidation, Mummification, Resin, Viscera.

Procedia PDF Downloads 49

Authors: James Boag

Abstract:

The needs of people with a diagnosis of dementia and their carers and families are physical, psychosocial, and psychological and begin at the time of diagnosis. There is frequently a lack of emotional support and counselling. Care- giving support is required from the presentation of the first symptoms of dementia until death. Alzheimer's disease begins decades before the clinical symptoms begin to appear, and in many cases, it remains undiagnosed, or diagnosed too late for any possible interventions to have any effect. However, if an incorrect diagnosis is given, it may result in a person being treated, without effect, for a type of dementia they do not have and delaying the interventions they should have received. Being diagnosed with dementia can cause emotional distress to the person, and physical and emotional support is needed, which will become more important as the disease progresses. The severity of the patient's dementia and their symptoms has a bearing of the impact on the carer and the support needed. A lack of insight and /or a denial of the diagnosis, grief, reacting to anticipated future losses, and coping methods to maximise the disease outcome, are things that should be addressed. Because of the stigma, it is important for carers not to lose contact with family and others because social isolation leads to depression and burnout. The impact on a carer's well- being and quality of life can be influenced by the severity of the illness, its type of dementia, its symptoms, healthcare support, financial and social status, career, age, health, residential setting, and relationship to the patient. Carer burnout due to lack of support leads to people diagnosed with dementia being put into residential care prematurely. Often dementia is not recognised as a terminal illness, limiting the ability of the person diagnosed with dementia and their carers to work on advance care planning and getting access to palliative and other support. Many carers have been satisfied with the physical support they were given in their everyday life, however, it was agreed that there was an immense unmet need for psychosocial support, especially after diagnosis and approaching end of life. Providing continuity and coordination of care is important. Training is necessary for providers to understand that every case is different, and they should understand the complexities. Grief, the emotional response to loss, is suffered during the progression of the disease and long afterwards, and carers should continue to be supported after the death of the person they were caring for.

Keywords: dementia, caring, challenges, needs

Procedia PDF Downloads 65

Authors: Syeda Hira, Muhammad Gulfraz

Abstract:

Background: Liver diseases cause serious health issues. Plants contain active compounds that significantly help in the treatment of various diseases. Ficus carica is traditionally used for the treatment of liver diseases. The purpose of the present study was the isolation and identification of active components from F.carica leaves which are responsible for hepatoprotective activity. Methods: The study was designed to identify the most active hepatoprotective sub-fraction from ethyl acetate fraction of Ficus carica by in vitro study and evaluation of its in vivo hepatoprotective effect in animal models. Ethyl acetate fraction was subjected to column, and a total of eight sub-fractions were obtained. In vitro, the hepatoprotective effect of all sub-fractions was determined on HepG2 cell lines. Toxicity was induced by CCl₄ (Carbon tetrachloride), and silymarin was used as a positive control. On the basis of the results, the most active sub-fraction was subjected to LC-MS and FT-IR analysis for the identification of bioactive compounds. In vivo, the hepatoprotective effect was determined in mice. Toxicity was induced by CCl₄; at the end of the experiment, biochemical parameters such as ALT, AST, ALP, bilirubin, and total protein were estimated in serum. Histopathology of liver tissues was also done. Results: Sub-fraction FVI exhibited significant (P<0.05) hepatoprotective activity as compared to other sub-fractions, which was almost similar to the standard drug silymarin. Six known bioactive compounds were identified from this sub-fraction after LC-MS analysis. In vivo, the hepatoprotective activity of sub-fraction FVI was evaluated in CCl₄-induced toxicated mice. Administration of CCl₄ significantly increased level of ALT (Alanine transaminase), AST (Aspartate aminotransferase), ALP (Alkaline phosphatase), and bilirubin and decreased the total protein. Treatment with sub-fraction FVI significantly (p<0.05) reversed the level of these biomarkers toward normal at both doses of 25 mg/kg and 50 mg/kg. Conclusion: Our findings confirmed the hepatoprotective effect of ethyl acetate fraction of F.carica. It could be a good candidate for the development of a natural hepatoprotective drug; pre-clinical investigation on ethyl acetate fraction is recommended.

Keywords: Ficus carica, hepatoprotective, CCl₄, bioactive compounds, liver markers

Procedia PDF Downloads 32

Authors: H. Alijani, M. Jabari, S. Matroodi, H. Zolqarnein, A. Sharafi, I. Zamani

Abstract:

Actinomycetes, Gram-positive bacteria, are interesting as a main producer of secondary metabolites and are important industrially and pharmaceutically. The marine environment is a potential source for new actinomycetes, which can provide novel bioactive compounds and industrially important enzymes. The aims of this study were to isolate and identify novel actinomycetes from Persian Gulf sediments and screen these isolates for the production of secondary metabolites, especially antibiotics, Using phylogenetic (16S rRNA gene sequence), morphological and biochemical analyses. 15 different actinomycete strains from Persian Gulf sediments at a depth of 5-10 m were identified. DNA extraction was done using Cinnapure DNA Kit. PCR amplification of 16S rDNA gene was performed using F27 and R1492 primers. Phylogenetic tree analysis was performed using the MEGA 6 software. Most of the isolated strains belong to the genus namely Streptomyces (14), followed by Nocardiopsis (1). Antibacterial assay of the isolates supernatant was performed using a standard disc diffusion assay with replication (n=3). The results of disk diffusion assay showed that most active strain against Proteus volgaris and Bacillus cereus was AMJ1 (16.46±0.2mm and 13.78±0.2mm, respectively), against Salmonella sp. AMJ7 was the most effective strain (10.13±0.2mm), and AMJ1 and AHA5 showed more inhibitory activity against Escherichia coli (8.04±0.02 mm and 8.2±0.03 ). The AMJ6 strain showed best antibacterial activity against Klebsiella sp. (8.03±0.02mm). Antifungal activity of AMJ2 showed that it was most active strain against complex (16.05±0.02mm) and against Aspergillus flavus strain AMJ1 was most active strain (16.4±0.2mm) and highest antifungal activity against Trichophyton mentagrophytes, Microsporum gyp serum and Candida albicans, were shown by AHA1 (21.03±0.02mm), AHA3 and AHA7 (18±0.03mm), AMJ6 (21.03±0.2mm) respectively. Our results revealed that the marine actinomycetes of Persian Gulf sediments were potent source of novel antibiotics and bioactive compounds and indicated that the antimicrobial metabolites were extracellular. Most of the secondary metabolites and antibiotics are extracellular in nature and extracellular products of actinomycetes show potent antimicrobial activities.

Keywords: antibacterial activity, antifungal activity, marine actinomycetes, Persian Gulf

Procedia PDF Downloads 271

Authors: S. Junaid S. Qazi, Denice C. Bay, Raymond Chew, Raymond J. Turner

Abstract:

EmrE, a member of the small multidrug resistance protein family in bacteria is considered to be the archetypical member of its family. It confers host resistance to a wide variety of quaternary cation compounds (QCCs) driven by proton motive force. Generally, purification yield is a challenge in all membrane proteins because of the difficulties in their expression, isolation and solubilization. EmrE is extremely hydrophobic which make the purification yield challenging. We have purified EmrE protein using two different approaches: organic solvent membrane extraction and hexahistidine (his6) tagged Ni-affinity chromatographic methods. We have characterized changes present between ligand affinity of untagged and his6-tagged EmrE proteins in similar membrane mimetic environments using biophysical experimental techniques. Purified proteins were solubilized in a buffer containing n-dodecyl-β-D-maltopyranoside (DDM) and the conformations in the proteins were explored in the presence of four QCCs, methyl viologen (MV), ethidium bromide (EB), cetylpyridinium chloride (CTP) and tetraphenyl phosphonium (TPP). SDS-Tricine PAGE and dynamic light scattering (DLS) analysis revealed that the addition of QCCs did not induce higher multimeric forms of either proteins at all QCC:EmrE molar ratios examined under the solubilization conditions applied. QCC binding curves obtained from the Trp fluorescence quenching spectra, gave the values of dissociation constant (Kd) and maximum specific one-site binding (Bmax). Lower Bmax values to QCCs for his6-tagged EmrE shows that the binding sites remained unoccupied. This lower saturation suggests that the his6-tagged versions provide a conformation that prevents saturated binding. Our data demonstrate that tagging an integral membrane protein can significantly influence the protein.

Keywords: small multidrug resistance (SMR) protein, EmrE, integral membrane protein folding, quaternary ammonium compounds (QAC), quaternary cation compounds (QCC), nickel affinity chromatography, hexahistidine (His6) tag

Procedia PDF Downloads 350

Authors: Neslihan Taskale Karatug, Mustafa Akcelik

Abstract:

Some literature data show that misL protein play a role on host immune response formed against Salmonella Typhimurium. The aim of the present study is to learn the role of the protein in S. Typhimurium pathogenicity. To describe certain functions of the protein, primarily recombinant misL protein was produced and purified. PCR was performed using a primer set targeted to passenger domain of the misL gene on S. Typhimurium LT2 genome. Amplicon and pet28a vector were enzymatically cleaved with EcoRI and NheI. The digested DNA materials were purified with High Pure PCR Product Purification Kit. The ligation reaction was achieved with the pure products. After preparation of competent Escherichia coli Dh5α, ligation mix was transformed into the cell by electroporation. To confirm the existence of insert gene, recombinant plasmid DNA of Dh5α was isolated with high pure plasmid DNA kit. Proved the correctness of recombinant plasmid was electroporated to BL21. The cell was induced by IPTG. After induction, the presence of recombinant protein was checked by SDS-PAGE. The recombinant misL protein was purified using HisPur Ni-NTA spin colon. The pure protein was shown by SDS-PAGE and western blot immünoassay. The concentration of the protein was measured BCA Protein Assay kit. In the wake of ligation with digested products (2 kb misL and 5.4 kb pet28a) visualised on gel size of the band was about 7.4 kb and was named as pNT01. The pNT01 recombinant plasmid was transformed into Dh5α and colonies were chosen in selective medium. Plasmid DNA isolation from them was carried out. PCR was achieved on the pNT01 to check misL and 2 kb band was observed on the agarose gel. After electroporation of the plasmid and induction of the cell, 68 kDa misL protein was seen. Subsequent to the purification of the protein, only a band was observed on SDS-PAGE. Association of the pure protein with anti-his antibody was verified by the western blot assay. The concentration of the pure misL protein was determined as 345 μg/mL. Production of polyclonal antibody will be achieved by using the obtained pure recombinant misL protein as next step. The role of the protein will come out on the immune system together some assays.

Keywords: cloning, Escherichia coli, recombinant protein purification, Salmonella Typhimurium

Procedia PDF Downloads 363

Authors: John Masani Nduko, Joseph Wafula Matofari

Abstract:

Many spontaneously fermented milk products are produced in Kenya, where they are integral to the human diet and play a central role in enhancing food security and income generation via small-scale enterprises. Fermentation enhances product properties such as taste, aroma, shelf-life, safety, texture, and nutritional value. Some of these products have demonstrated therapeutic and probiotic effects although recent reports have linked some to death, biotoxin infections, and esophageal cancer. These products are mostly processed from poor quality raw materials under unhygienic conditions resulting to inconsistent product quality and limited shelf-lives. Though very popular, research on their processing technologies is low, and none of the products has been produced under controlled conditions using starter cultures. To modernize the processing technologies for these products, our study aims at describing the microbiology and biochemistry of a representative Kenyan spontaneously fermented milk product, Mursik using modern biotechnology (DNA sequencing) and their chemical composition. Moreover, co-creation processes reflecting stakeholders’ experiences on traditional fermented milk production technologies and utilization, ideals and senses of value, which will allow the generation of products based on common ground for rapid progress will be discussed. Knowledge of the value of clean starting raw material will be emphasized, the need for the definition of fermentation parameters highlighted, and standard equipment employment to attain controlled fermentation discussed. This presentation will review the available information regarding traditional fermented milk (Mursik) and highlight our current research work on the application of molecular approaches (metagenomics) for the valorization of Mursik production process through starter culture/ probiotic strains isolation and identification, and quality and safety aspects of the product. The importance of the research and future research areas on the same subject will also be highlighted.

Keywords: lactic acid bacteria, high throughput biotechnology, spontaneous fermentation, Mursik

Procedia PDF Downloads 263