Search results for: blood cultures

Authors: Laura Villamizar, David Wright, Claudia Baena, Marie Foxwell, Maureen O'Callaghan

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Leguminous crops can be self-sufficient for their nitrogen requirements when their roots are nodulated with an effective Rhizobium strain and for this reason seed or soil inoculation is practiced worldwide to ensure nodulation and nitrogen fixation in grain and forage legumes. The most widely used method of applying commercially available inoculants is using peat cultures which are coated onto seeds prior to sowing. In general, rhizobia survive well in peat, but some species die rapidly after inoculation onto seeds. The development of improved formulation methodology is essential to achieve extended persistence of rhizobia on seeds, and improved efficacy. Formulations could be solid or liquid. Most popular solid formulations or delivery systems are: wettable powders (WP), water dispersible granules (WG), and granules (DG). Liquid formulation generally are: suspension concentrates (SC) or emulsifiable concentrates (EC). In New Zealand, R. leguminosarum bv. trifolii strain TA1 has been used as a commercial inoculant for white clover over wide areas for many years. Seeds inoculation is carried out by mixing the seeds with inoculated peat, some adherents and lime, but rhizobial populations on stored seeds decline over several weeks due to a number of factors including desiccation and antibacterial compounds produced by the seeds. In order to develop a more stable and suitable delivery system to incorporate rhizobia in pastures, two strains of R. leguminosarum (TA1 and CC275e) and several formulations and processes were explored (peat granules, self-sticky peat for seed coating, emulsions and a powder containing spray dried microcapsules). Emulsions prepared with fresh broth of strain TA1 were very unstable under storage and after seed inoculation. Formulations where inoculated peat was used as the active ingredient were significantly more stable than those prepared with fresh broth. The strain CC275e was more tolerant to stress conditions generated during formulation and seed storage. Peat granules and peat inoculated seeds using strain CC275e maintained an acceptable loading of 108 CFU/g of granules or 105 CFU/g of seeds respectively, during six months of storage at room temperature. Strain CC275e inoculated on peat was also microencapsulated with a natural biopolymer by spray drying and after optimizing operational conditions, microparticles containing 107 CFU/g and a mean particle size between 10 and 30 micrometers were obtained. Survival of rhizobia during storage of the microcapsules is being assessed. The development of a stable product depends on selecting an active ingredient (microorganism), robust enough to tolerate some adverse conditions generated during formulation, storage, and commercialization and after its use in the field. However, the design and development of an adequate formulation, using compatible ingredients, optimization of the formulation process and selecting the appropriate delivery system, is possibly the best tool to overcome the poor survival of rhizobia and provide farmers with better quality inoculants to use.

Keywords: formulation, Rhizobium leguminosarum, storage stability, white clover

Procedia PDF Downloads 135

Authors: Meilan Jin, Mary Jane Moran

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This study aims to introduce the framework of culturally responsive home visiting (CRHV) to head start teacher professional sessions in the Southeastern of the US and investigate its influence on the evolving beliefs of teachers about their roles and relationships with families in-home visits. The framework orients teachers to an effective way of taking on the role of learner to listen for spoken and unspoken needs and look for family strengths. In addition, it challenges the deficit model that is grounded on 'cultural deprivation,' and it stresses the value of family cultures and advocates equal, collaborative parent-teacher relationships. The home visit reflection papers and focus group transcriptions of eight teachers have been collected since 2010 throughout a five-year longitudinal collaboration with them. Reflection papers were written by the teachers before and after introducing the CRHV framework, including the details of visit purposes and actions and their plans for later home visits. Particularly, the CRHV framework guided the teachers to listen and look for information about family-living environments; parent-child interactions; child-rearing practices; and parental beliefs, values, and needs. Two focus groups were organized in 2014 by asking the teachers to read their written reflection papers and then discussing their shared beliefs and experiences of home visits in recent years. The average length of the discussions was one hour, and the discussions were audio-recorded and transcribed verbatim. Moreover, the data were analyzed using constant comparative analysis, and the analysis was verified through (a) the uses of multiple data sources, (b) the involvement of multiple researchers, (c) coding checks, and (d) the provisions of the thick descriptions of the findings. The study findings corroborate that the teachers become to reposition themselves as 'knowledge seekers' through reorienting their cynosure toward 'setting stones' to learn, grow, and change rather than framing their home visits. The teachers also continually engage in careful listening, observing, questioning, and dialoguing, and these actions reflect their care toward parents. The value of teamwork with parents is advocated, and the teachers recognize that when parents feel empowered, they are active and committed to doing more for their children, which can further advantage proactive long-term parent-teacher collaborations. The study findings also validate that the framework is influential for educators to provide the experiences of home visiting that is culturally responsive and to share collaborative relationships with caregivers. The long-term impact of the framework further implies that teachers continue to put themselves in the position of evolving, including beliefs and actions, to better work with children and families who are culturally, ethnically, and linguistically different from them. This framework can be applicable to educators and professionals who are looking for avenues to bridge the relationship between home and school and parents and teachers.

Keywords: culturally responsive home visit, early childhood education, parent–teacher collaboration, teacher professional development

Procedia PDF Downloads 82

Authors: Laura G. Caixeta, Maico R. Severino

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One of the biggest challenges that the industries find nowadays is to incorporate sustainability practices into its operations. The Green Supply Chain Management (GSCM) concept assists industries in such incorporation. For the full application of this concept is important that enterprises of a same supply chain have the GSCM practices coordinated among themselves. Note that this type of analyses occurs on the context of developed countries and sectors considered big impactors (as automotive, mineral, among others). The propose of this paper is to analyze as the GSCM concepts are applied on the Brazilian animal nutrition industries. The method used was the Case Study. For this, it was selected a supply chain relationship composed by animal nutrition products manufacturer (Enterprise A) and its supplier of animal waste, such as blood, viscera, among others (Enterprise B). First, a literature review was carried out to identify the main GSCM practices. Second, it was done an individual analysis of each one selected enterprise of the application of GSCM concept. For the observed practices, the coordination of each practice in this supply chain was studied. And, it was developed propose of GSCM applications for the practices no observed. The findings of this research were: a) the systematization of main GSCM practices, as: Internal Environment Management, Green Consumption, Green Design, Green Manufacturing, Green Marketing, Green Packaging, Green Procurement, Green Recycling, Life Cycle Analysis, Consultation Selection Method, Environmental Risk Sharing, Investment Recovery, and Reduced Transportation Time; b) the identification of GSCM practices on Enterprise A (7 full application, 3 partial application and 3 no application); c) the identification of GSCM practices on Enterprise B (2 full application, 2 partial application and 9 no application); d) the identification of how is the incentive and the coordination of the GSCM practices on this relationship by Enterprise A; e) proposals of application and coordination of the others GSCM practices on this supply chain relationship. Based on the study, it can be concluded that its possible apply GSCM on animal nutrition industries, and when occurs the motivation on the application of GSCM concepts by a supply chain echelon, these concepts are deployed for the others supply chain echelons by the coordination (orchestration) of the first echelon.

Keywords: animal nutrition industries, coordination, green supply chain management, supply chain management, sustainability

Procedia PDF Downloads 113

Authors: Lavrentiadou S. N., Angelou V., Chatzimisios K., Papazoglou L.

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The aim of this study was the isolation and culture of keratinocytes and fibroblasts from feline skin to ultimately create an artificial engineered skin (including dermis and epidermis) useful for the effective treatment of large cutaneous deficits in cats. Epidermal keratinocytes and dermal fibroblasts were freshly isolated from skin biopsies using an 8 mm biopsy punch obtained from 8 healthy cats that had undergone ovariohysterectomy. The owner’s consent was obtained. All cats had a complete blood count and a serum biochemical analysis and were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) preoperatively. The samples were cut into small pieces and incubated with collagenase (2 mg/ml) for 5-6 hours. Following digestion, cutaneous cells were filtered through a 100 μm cell strainer, washed with DMEM, and grown in DMEM supplemented with 10% FBS. The undigested epidermis was washed with DMEM and incubated with 0.05% Trypsin/0.02% EDTA (TE) solution. Keratinocytes recovered in the TE solution were filtered through a 100 μm and a 40 μm cell strainer and, following washing, were grown on a collagen type I matrix in DMEM: F12 (3:1) medium supplemented with 10% FΒS, 1 μm hydrocortisone, 1 μm isoproterenol and 0.1 μm insulin. Both fibroblasts and keratinocytes were grown in a humidified atmosphere with 5% CO2 at 37oC. The medium was changed twice a week and cells were cultured up to passage 4. Cells were grown to 70-85% confluency, at which point they were trypsinized and subcultured in a 1:4 dilution. The majority of the cells in each passage were transferred to a freezing medium and stored at -80oC. Fibroblasts were frozen in DMEM supplemented with 30% FBS and 10% DMSO, whereas keratinocytes were frozen in a complete keratinocyte growth medium supplemented with 10% DMSO. Both cell types were thawed and successfully grown as described above. Therefore, we can create a bank of fibroblasts and keratinocytes, from which we can recover cells for further culture and use for the generation of skin equivalent in vitro. In conclusion, cutaneous cell isolation and cell culture and expansion were successfully developed. To the authors’ best knowledge, this is the first study reporting isolation and culture of keratinocytes and fibroblasts from feline skin. However, these are preliminary results and thus, the development of autologous-engineered feline skin is still in process.

Keywords: cat, fibroblasts, keratinocytes, skin equivalent, wound

Procedia PDF Downloads 87

Authors: Luna Degife, Desalegn Belay, Yoseph Worku, Tigist Tesfaye, Assefa Tufa, Abyot Bekele, Zegeye Hailemariam, Abay Hagos

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Half of the world’s population is at risk of Dengue Fever (DF), a highly under-recognized and underreported mosquito-borne viral disease with high prevalence in the tropical and subtropical regions. Globally, an estimated 50 to 200 million cases and 20, 000 DF deaths occur annually as per the world health organization report. In Ethiopia, the first outbreak occurred in 2013 in Diredawa administration city. Afterward, three outbreaks have been reported from the eastern part of the country. We received a report of the fifth DF outbreak for Ethiopia and the second for Diredawa city on October 4, 2015. We conducted the investigation to confirm the outbreak, identify the risk factors for the repeatedly occurrence of the disease and implement control measures. We conducted un- matched case-control study and defined a suspected DF case as any person with fever of 2-7 days and 2 or more of the following: a headache, arthralgia, myalgia, rash, or bleeding from any part of the body. Controls were residents of Diredawa city without DF symptoms. We interviewed 70 Cases and 140 controls from all health facilities in Diredawa city from October 7 to 15; 2015. Epi Info version 7.1.5.0 was used to analyze the data and multivariable logistic regression was conducted to assess risk factors for DF. Sixty-nine blood samples were collected for Laboratory confirmation.The mean age for cases was 23.7±9.5 standard deviation (SD) and for controls 31.2±13 SD. Close contact with DF patient (Adjusted odds ratio (AOR)=5.36, 95% confidence interval(CI): 2.75-10.44), nonuse of long-lasting insecticidal nets (AOR=2.74, 95% CI: 1.06-7.08) and availability of stagnant water in the village (AOR=3.61, 95% CI:1.31-9.93) were independent risk factors associated with higher rates of the disease. Forty-two samples were tested positive. Endemicity of DF is becoming a concern for Diredawa city after the first outbreak. Therefore, effective vector control activities need to be part of long-term preventive measures.

Keywords: dengue fever, Diredawa, outbreak, risk factors, second

Procedia PDF Downloads 251

Authors: Munkhtuul G., Bolortsetseg Z., Lutzul M., Sugar N., Nyamdorj D., Nomundari B., Zesemdorj O., Erdenebayar N., Lkhagvasuren T. S., Munkhbayarlakh S., Bayasgalan T. Uurtuya S. H.

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Background: Elevated serum uric acid (SUA) levels are observed in metabolic and cardiovascular conditions as an early predictor of metabolic syndrome (MS). Hyperuricemia, characterised by high uric acid levels in serum, increases the risk of developing MS by 1.6 times. Being overweight and obese significantly contributes to developing MS and cardiovascular disorders. In Mongolia, the prevalence of overweight and obesity is reaching 48.8% among individuals aged 15 to 49 years, indicating a potential surge in the incidence of MS, cardiovascular disorders, diabetes mellitus, and gout.Objective: This study aimed to determine the SUA levels in men diagnosed with MS and investigate the factors influencing these levels.Methods: A total of 119 men aged 30-60, who underwent preventive examinations and resided in Ulaanbaatar city, were included in the study. The criteria established by the International Diabetes Federation (IDF), American Heart Association (AHA), and the National Heart, Lung, and Blood Institute (NHLBI) were employed to define metabolic syndrome. Hyperuricemia was defined as SUA levels ≥7 mg/dL. Dietary intake was evaluated through the 24-hour recall method.Results: The study revealed that the prevalence of MS among the participants was 42.9% (n=51), with hyperuricemia observed in 16.8% (n=20) of the individuals. Among men diagnosed with MS, 21.3% (n=10) exhibited hyperuricemia. The mean SUA levels were as follows: 4.7±0.8 mg/dL in the healthy group, 5.9±1.1 mg/dL in men without MS but presenting central obesity, and 6.2±1.3 mg/dL in men with MS. After adjusting for age and body mass index (BMI), a positive correlation was observed between SUA levels and triglycerides (β=0.93) as well as lipid accumulation product (LAP) (β=0.92) in men with MS. In the central obesity group, SUA levels exhibited a positive correlation with triglycerides (β=0.91), visceral adiposity index (VAI) (β=0.73), LAP (β=0.92), and cardiometabolic index (CMI) (β=0.69). The risk of hyperuricemia increased by 3.29 times with elevated triglycerides and 3.53 times with an increased LAP.Conclusion: The findings indicate that abdominal fat accumulation, as indicated by elevated triglyceride levels and LAP, is associated with increased SUA levels in men with MS. However, no significant relationship was observed between SUA levels and dietary intake.

Keywords: central obesity, obesity, triglycerides, hyperuricemia

Procedia PDF Downloads 45

Authors: Caspar S. Carson, Gabriel W. Prather, Nicholas E. Wong, Jeffery R. Anton, William H. McCoy

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Cutibacterium acnes is a commensal bacterium found on human skin that has been linked to acne. C. acnes can also be an opportunistic pathogen when it infiltrates the body during surgery. This pathogen can cause dangerous infections of medical implants, such as shoulder replacements, leading to life-threatening blood infections. Compounding this issue, C. acnes resistance to many antibiotics has become an increasing problem worldwide, creating a need for special forms of treatment. C. acnes expresses the protein RoxP, and it requires this protein to colonize human skin. Though this protein is required for C. acnes skin colonization, its function is not yet understood. Inhibition of RoxP function might be an effective treatment for C. acnes infections. To develop such reagents, the McCoy Laboratory generated four unique anti-RoxP antibodies. Preliminary studies in the McCoy Lab have established that each antibody binds a distinct site on RoxP. To assess the potential of these antibodies as therapeutics, it is necessary to specifically characterize these antibody epitopes and evaluate them in assays that assess their ability to inhibit RoxP-dependent C. acnes growth. To provide material for these studies, an antibody expression construct, Fv-clasp(v2), was adapted to encode anti-RoxP antibody sequences. The author hypothesizes that this expression strategy can produce sufficient amounts of >95% pure antibody fragments for further characterization of these antibodies. Four anti-RoxP Fv-clasp(v2) expression constructs (pET vector-based) were transformed into E. coli BL21-Gold(DE3) cells and a small-scale expression and purification trial was performed for each construct to evaluate anti-RoxP Fv-clasp(v2) yield and purity. Successful expression and purification of these antibody constructs will allow for their use in structural studies, such as protein crystallography and cryogenic electron microscopy. Such studies would help to define the antibody binding sites on RoxP, which could then be leveraged in the development of certain methods to treat C. acnes infection through RoxP inhibition.

Keywords: structural biology, protein expression, infectious disease, antibody, therapeutics, E. coli

Procedia PDF Downloads 42

Authors: Abirami Baleswaran, Christel Couderc, Loubnah Belahcen, Jean Dayde, Hélène Tormo, Gwénaëlle Jard

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Cheeses are often reported to be spoiled by Pseudomonas spp., responsible for defects in appearance, texture, taste, and smell, leading to their non-marketing and even their destruction. Despite preventive actions, problems linked to Pseudomonas spp. are difficult to control by the lack of knowledge and control of these contaminants during the cheese manufacturing. Lactic goat cheese producers are not spared by this problem and are looking for solutions to decrease the number of spoiled cheeses. To explore different hypotheses, experiments are needed. However, cheese-making experiments at the pilot scale are expensive and time consuming. Thus, there is a real need to develop a miniature cheeses model system under controlled conditions. In a previous study, several miniature cheese models corresponding to different type of commercial cheeses have been developed for different purposes. The models were, for example, used to study the influence of milk, starters cultures, pathogen inhibiting additives, enzymatic reactions, microflora, freezing process on cheese. Nevertheless, no miniature model was described on the lactic goat cheese. The aim of this work was to develop a miniature cheese model system under controlled laboratory conditions which resembles commercial lactic goat cheese to study Pseudomonas spp. spoilage during the manufacturing and ripening process. First, a protocol for the preparation of miniature cheeses (3.5 times smaller than a commercial one) was designed based on the cheese factorymanufacturing process. The process was adapted from “Rocamadour” technology and involves maturation of pasteurized milk, coagulation, removal of whey by centrifugation, moulding, and ripening in a little scale cellar. Microbiological (total bacterial count, yeast, molds) and physicochemical (pH, saltinmoisture, moisture in fat-free)analyses were performed on four key stages of the process (before salting, after salting, 1st day of ripening, and end of ripening). Factory and miniature cheeses volatilomewere also obtained after full scan Sift-MS cheese analysis. Then, Pseudomonas spp. strains isolated from contaminated cheeses were selected on their origin, their ability to produce pigments, and their enzymatic activities (proteolytic, lecithinasic, and lipolytic). Factory and miniature curds were inoculated by spotting selected strains on the cheese surface. The expression of cheese spoilage was evaluated by counting the level of Pseudomonas spp. during the ripening and by visual observation and under UVlamp. The physicochemical and microbiological compositions of miniature cheeses permitted to assess that miniature process resembles factory process. As expected, differences involatilomes were observed, probably due to the fact that miniature cheeses are made usingpasteurized milk to better control the microbiological conditions and also because the little format of cheese induced probably a difference during the ripening even if the humidity and temperature in the cellar were quite similar. The spoilage expression of Pseudomonas spp. was observed in miniature and factory cheeses. It confirms that the proposed model is suitable for the preparation of miniature cheese specimens in the spoilage study of Pseudomonas spp. in lactic cheeses. This kind of model could be deployed for other applications and other type of cheese.

Keywords: cheese, miniature, model, pseudomonas spp, spoilage

Procedia PDF Downloads 119

Authors: Obadire F. O., Obadire, S. O., Adeoti R. F., Pirgozliev V.

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An experiment was conducted to determine the efficacy of utilizing African black plum seed nut (ABPNBD) meal on hematological and biochemical indices of broiler chicken ration formulated to substitute wheat offal. A total of 150- 1day-old, male Agrited birds were reared for 28 days of the experiment. The birds were assigned to five dietary treatments, with ten birds per treatment replicated 3 times. Experimental diets were formulated by supplementing the milled African black plum nut at (0, 5, 10, 12.5, and 15%) inclusion levels in the starter broiler’s ratio designated as T1 (control diet containing no ABPBD), Treatments (T2, 3,4 and 5) contained ABPNBD at 5, 10, 12.5, and 15%, respectively, in a completely randomized design. The hematological and biochemical indices of the birds were determined. The result revealed that all hematological parameters measured were significant (P <0.05) except for WBC. Increasing inclusion levels of ABPNBD decreased the PCV, HB, and RBC of the birds across the treatment groups. Birds fed 12.5 and 15% ABPNBD diets recorded the least of the parameters. The result of the serum biochemical indices showed significant (P < 0.05) influence for all parameters measured except for alanine transaminase (ALT), (AST), and creatinine. The total protein (TP), albumin, globulin, and glucose values were reduced across the treatment group as ABPNBD inclusion increased. Birds fed above 10% ABPNBD recorded the lowest value of TP, albumin, globulin, and glucose when compared with birds on a control diet and other treatments. The uric acid ranged from 3.85 to 2 .13 mmol/L, while creatinine ranged from 62.00 to 53.50 mmol/l. AST ranged between 8.50 u/l (5%) to 7.90 u/l (10%). ALT ranged between 7.50 u/l (12.5%) to 5.50 u/l (5 and 10%). In conclusion, dietary inclusion of African black plum up to 10% has no detrimental effect on the health of the starter chickens. Meanwhile, inclusion above 10% revealed a negative effect on some blood parameters measured. Therefore, African black plum should be supplemented with probable probiotics or subjected to different processing methods if to be used at a 15% inclusion level for optimal results.

Keywords: African black plum seed, starter broiler chickens, hematological and serum biochemical indices, (Vitex doniana)

Procedia PDF Downloads 27

Authors: D. F. Zeynalov, T. V. Kartseva, O. V. Sorokin

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Currently, approaches to assess cardiointervalography are based on the calculation of data variance intervals RR. However, they do not allow the evaluation of features related to a period of the cardiac cycle, so how electromechanical phenomena during cardiac subphase are characterized by differently directed changes. Therefore, we have proposed a method of subphase analysis of the cardiac cycle, developed in the department of hominal physiology Novosibirsk State Medical University to identify the features of the dispersion subphase of the cardiac cycle. In the present paper we have examined the 5-minute intervals cardiointervalography (CIG) to isolate RR-, QT-, ST-ranges in healthy children and children with acute respiratory diseases (ARD) in comparison. It is known that primary school-aged children suffer at ARD 5-7 times per year. Consequently, it is one of the most relevant problems in pediatrics. It is known that the spectral indices and indices of temporal analysis of heart rate variability are highly sensitive to the degree of intoxication during immunological process. We believe that the use of subphase analysis of heart rate will allow more thoroughly evaluate responsiveness of the child organism during the course of ARD. The study involved 60 primary school-aged children (30 boys and 30 girls). In order to assess heart rhythm regulation, the record CIG was used on the "VNS-Micro" device of Neurosoft Company (Ivanovo) for 5 minutes in the supine position and 5 minutes during active orthostatic test. Subphase analysis of variance QT-interval and ST-segment was performed on the "KardioBOS" software Biokvant Company (Novosibirsk). In assessing the CIG in the supine position and in during orthostasis of children with acute respiratory diseases only RR-intervals are observed typical trend of general biological reactions through pressosensitive compensation mechanisms to lower blood pressure, but compared with healthy children the severity of the changes is different, of sick children are more pronounced indicators of heart rate regulation. But analysis CIG RR-intervals and analysis subphase ST-segment have yielded conflicting trends, which may be explained by the different nature of the intra- and extracardiac influences on regulatory mechanisms that implement the various phases of the cardiac cycle.

Keywords: acute respiratory diseases, cardiointervalography, subphase analysis, cardiac cycle

Procedia PDF Downloads 255

Authors: Hamza I. Isa, Gezina C. H. Ferreira, Jan E. Crafford, Christoffel J. Botha

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Moraea pallida Bak. (yellow tulip) poisoning is the most important plant-induced cardiac glycoside toxicosis in South Africa. Cardiac glycoside poisonings collectively account for about 33 and 10 % mortalities due to plants, in large and small stock respectively, in South Africa. The toxic principle is 1α, 2α-epoxyscillirosidine, a bufadienolide. The aim of the study was to investigate the potential to develop a vaccine against epoxyscillirosidine. Epoxyscillirosidine and the related bufadienolides proscillaridin and bufalin, which are commercially available, were conjugated to the carrier proteins [Hen ovalbumin (OVA), bovine serum albumin (BSA) and keyhole limpet haemocyanin (KLH)], rendering them immunogenic. Adult male New Zealand White rabbits were immunized. In Trials 1 and 2, rabbits (n=6) were, each assigned to two groups. Experimental animals (n=3; n=4) were vaccinated with epoxyscillirosidine-OVA conjugate, while the control (n=3; n=2) were vaccinated with OVA, using Freund’s complete and incomplete and Montanide adjuvants, for Trials 1 and 2, respectively. In Trial 3, rabbits (n=15), randomly allocated to 5 equal groups (I, II, III, IV and V), were vaccinated with proscillaridin-BSA, bufalin-BSA, epoxyscillirosidine-KLH, epoxyscillirosidine-BSA conjugates, and BSA respectively, using Montanide as adjuvant. Vaccination was on Days 0, 21 and 42. Additional vaccinations were done on Day 56 and 63 for Trial 1. Vaccination was by intradermal injection of 0.4 ml of the immunogen (4 mg/ml [Trial 1] and 8 mg/ml for Trials 2 and Trial 3, respectively). Blood was collected pre-vaccination and at 3 week intervals following each vaccination. Antibody response was determined using an indirect ELISA. There was poor immune response associated with the dose (0.4 mg per rabbit) and adjuvant used in Trial 1. Antibodies were synthesized against the conjugate administered in Trial 2. For Trail 3, antibodies against the immunogens were successfully raised in rabbits with epoxyscillirosidine-KLH inducing the highest immune response. The antibodies raised against proscillaridin and bufalin cross-reacted with epoxyscillirosidine when used as antigen in the ELISA. The study successfully demonstrated the synthesis of antibodies against the bufadienolide conjugates administered. The cross-reactivity of proscillaridin and bufalin with epoxyscillirosidine could potentially be utilized as alternative to epoxyscillirosidine in future studies to prevent yellow tulp poisoning by vaccination.

Keywords: antibodies , bufadienolides, cross-reactivity, epoxyscillirosidine, Moraea pallida, poisoning

Procedia PDF Downloads 139

Authors: Kayla Belanger, Pascale Vigneron, Guy Schlatter, Bernard Devauchelle, Christophe Egles

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A severe injury to a peripheral nerve leads to its degeneration and the loss of sensory and motor function. To this day, there still lacks a more effective alternative to the autograft which has long been considered the gold standard for nerve repair. In order to overcome the numerous drawbacks of the autograft, tissue engineered biomaterials may be effective alternatives. Silk fibroin is a favorable biomaterial due to its many advantageous properties such as its biocompatibility, its biodegradability, and its robust mechanical properties. In this study, bio-mimicking multi-channeled nerve guidance conduits made of aligned nanofibers achieved by electrospinning were functionalized with signaling biomolecules and were tested in vitro and in vivo for nerve regeneration support. Silk fibroin (SF) extracted directly from silkworm cocoons was put in solution at a concentration of 10wt%. Poly(ethylene oxide) (PEO) was added to the resulting SF solution to increase solution viscosity and the following three electrospinning solutions were made: (1) SF/PEO solution, (2) SF/PEO solution with nerve growth factor and ciliary neurotrophic factor, and (3) SF/PEO solution with nerve growth factor and neurotrophin-3. Each of these solutions was electrospun into a multi-layer architecture to obtain mechanically optimized aligned nanofibrous mats. For in vitro studies, aligned fibers were treated to induce β-sheet formation and thoroughly rinsed to eliminate presence of PEO. Each material was tested using rat embryo neuron cultures to evaluate neurite extension and the interaction with bio-functionalized or non-functionalized aligned fibers. For in vivo studies, the mats were rolled into 5mm long multi-, micro-channeled conduits then treated and thoroughly rinsed. The conduits were each subsequently implanted between a severed rat sciatic nerve. The effectiveness of nerve repair over a period of 8 months was extensively evaluated by cross-referencing electrophysiological, histological, and movement analysis results to comprehensively evaluate the progression of nerve repair. In vitro results show a more favorable interaction between growing neurons and bio-functionalized silk fibers compared to pure silk fibers. Neurites can also be seen having extended unidirectionally along the alignment of the nanofibers which confirms a guidance factor for the electrospun material. The in vivo study has produced positive results for the regeneration of the sciatic nerve over the length of the study, showing contrasts between the bio-functionalized material and the non-functionalized material along with comparisons to the experimental control. Nerve regeneration has been evaluated not only by histological analysis, but also by electrophysiological assessment and motion analysis of two separate natural movements. By studying these three components in parallel, the most comprehensive evaluation of nerve repair for the conduit designs can be made which can, therefore, more accurately depict their overall effectiveness. This work was supported by La Région Picardie and FEDER.

Keywords: electrospinning, nerve guidance conduit, peripheral nerve regeneration, silk fibroin

Procedia PDF Downloads 226

Authors: S. Kozlovski, S.W. Feigelson, R. Alon

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The b2 integrin ligands ICAM-1 ICAM-2 and the endothelial VLA-4 integrin ligand VCAM-1 are constitutively expressed on different lung vessels and on high endothelial venules (HEVs), the main portal for lymphocyte entry from the blood into lung draining lymph nodes. ICAMs are also ubiquitously expressed by many antigen-presenting leukocytes and have been traditionally suggested as critical for the various antigen-specific immune synapses generated by these distinct leukocytes and specific naïve and effector T cells. Loss of both ICAM-1 and ICAM-2 on the lung vasculature reduces the ability to patrol monocytes and Tregs to patrol the lung vasculature at a steady state. Our new findings suggest, however, that in terms of innate leukocyte trafficking into the lung lamina propria, both constitutively expressed and virus-induced vascular VCAM-1 can functionally compensate for the loss of these ICAMs. In a mouse model for influenza infection, neutrophil and NK cell recruitment and clearance of influenza remained normal in mice deficient in both ICAMs. Strikingly, mice deficient in both ICAMs also mounted normal influenza-specific CD8 proliferation and differentiation. In addition, these mice normally combated secondary influenza infection, indicating that the presence of ICAMs on conventional dendritic cells (cDCs) that present viral antigens are not required for immune synapse formation between these APCs and naïve CD8 T cells as previously suggested. Furthermore, long-lasting humoral responses critical for protection from a secondary homosubtypic influenza infection were also normal in mice deficient in both ICAM-1 and ICAM-2. Collectively, our results suggest that the expression of ICAM-1 and ICAM-2 on lung endothelial and epithelial cells, as well as on DCs and B cells, is not critical for the generation of innate or adaptive anti-viral immunity in the lungs. Our findings also suggest that endothelial VCAM-1 can substitute for the functions of vascular ICAMs in leukocyte trafficking into various lung compartments.

Keywords: emigration, ICAM-1, lymph nodes, VCAM-1

Procedia PDF Downloads 112

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

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Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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Authors: Salwa S. Dawaki, Init Ithoi, Sa’adatu I. Yelwa

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Introduction: Parasitic infections are major public health problems worldwide, particularly in developing countries. Two third of the world population is infected while about 3 billion are at risk of parasitic infections. It is demonstrated that most parasitic infections occur as multiple infections especially among poor and rural communities of most countries in the tropical regions. Parasitic infections are endemic in Nigeria, yet multiple infections are rarely reported. The study aimed to estimate the prevalence and identify factors associating with multiple parasitic infections among rural population in Kano State Nigeria. Methodology: A cross-sectional survey was conducted from June to August 2013 in rural Kano State, Nigeria. Three samples stool, urine, and blood were collected from each of the 551 volunteers aged between one and ninety years old recruited for the survey. A pre-tested questionnaire was used to obtain epidemiological data. Data were analysed using appropriate descriptive, univariate and multivariate logistic regression methods. Major findings: The participants were 61.7% male, 38.3% female, and 69.0% were adults of 15 years and above. Overall, 463 (84%) were infected with parasitic infections among which 60.9% had multiple infections. A total of 15 parasitic species were recovered, and up to 8 different parasitic species were found concurrently in a single host. Plasmodium was the most common parasite followed by Blastocystis, Entamoeba species, and hookworms. It was found that presence of an infected family member (P = 0.017; OR = 1.52; 95% CI = 1.08, 2.13) and not wearing shoes outside home (P = 0.043; OR = 1.50; 95% CI = 1.01, 2.18) significantly associated with higher risk of having multiple parasitic infections among the studied population. Conclusion: Parasitic infections pose a public health challenge in the rural community of Kano. Multiple parasitic infections are highly prevalent and presence of an infected family member as well as not wearing proper foot wear outside home increases the risk of infection. Poor hygiene, unfavourable socioeconomic conditions, and culture promote survival and transmission of parasites. There is a need for implementation of integrated approach aimed at controlling or eliminating the infections with emphasis on public awareness.

Keywords: multiple infections, parasitic infections, poor hygiene, risk of infection

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Authors: Sadia Munir

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The aim of this study is to investigate changes in placental development and vascular dysfunction which subsequently affect feto-maternal health in pregnancies complicated by gestational diabetes mellitus (GDM). Fetal and postnatal adverse health outcomes of GDM are shown to be associated with disturbances in placental structure and function. Children of women with GDM are more likely to be obese and diabetic in childhood and adulthood. GDM also increases the risk of adverse pregnancy outcomes, including preeclampsia, birth injuries, macrosomia and neonatal hypoglycemia, respiratory distress syndrome, neonatal cardiac dysfunction and stillbirth. Incidences of type 2 diabetes in the MENA region are growing at an alarming rate which is estimated to become more than double by 2030. Five of the top 10 countries for diabetes prevalence in 2010 were in the Gulf region. GDM also increases the risk of development of type 2 diabetes. Interestingly, more than half of the women with GDM develop diabetes later in their life. The human placenta is a temporary organ located at the interface between mother and fetal blood circulation. Placenta has a central role as both a producer as well as a target of several molecules that are involved in placental development and function. We have investigated performed a Pubmed search with key words placenta, GDM, placental villi, vascularization, cytokines, growth factors, inflammation, hypoxia, oxidative stress and pathophysiology. We have investigated differences in the development and vascularization of placenta, their underlying causes and impact on feto-maternal health through literature review. We have also identified gaps in the literature and research questions that need to be answered to completely understand the central role of placenta in the GDM. This study is important in understanding the pathophysiology of placenta due to changes in the vascularization of villi, surface area and diameter of villous capillaries in pregnancies complicated by GDM. It is necessary to understand these mechanisms in order to develop treatments to reverse their effects on placental malfunctioning, which in turn, will result in improved mother and child health.

Keywords: gestational diabetes mellitus, placenta, vasculature, villi

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Authors: A. V. Sokolov, S. V. Vorobyev, A. Yu. Efimtcev, V. Yu. Lobzin, I. A. Lupanov, O. A. Cherdakov, V. A. Fokin

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Alzheimer’s disease (AD) is the most common form of dementia. Different brain regions are involved to the pathological process of AD. The purpose of this study was to evaluate brain activation by visual memory task in patients with Alzheimer's disease and determine correlation between memory impairment and atrophy of memory specific brain regions of frontal and medial temporal lobes. To investigate the organization of memory and localize cortical areas activated by visual memory task we used functional magnetic resonance imaging and to evaluate brain atrophy of patients with Alzheimer's disease we used voxel-based morphometry. FMRI was performed on 1.5 T MR-scanner Siemens Magnetom Symphony with BOLD (Blood Oxygenation Level Dependent) technique, based on distinctions of magnetic properties of hemoglobin. For test stimuli we used series of 12 not related images for "Baseline" and 12 images with 6 presented before for "Active". Stimuli were presented 3 times with reduction of repeated images to 4 and 2. Patients with Alzheimer's disease showed less activation in hippocampal formation (HF) region and parahippocampal gyrus then healthy persons of control group (p<0.05). The study also showed reduced activation in posterior cingulate cortex (p<0.001). Voxel-based morphometry showed significant atrophy of grey matter in Alzheimer’s disease patients, especially of both temporal lobes (fusiform and parahippocampal gyri); frontal lobes (posterior cingulate and superior frontal gyri). The study showed correlation between memory impairment and atrophy of memory specific brain regions of frontal and medial temporal lobes. Thus, reduced activation in hippocampal formation and parahippocampal gyri, in posterior cingulate gyrus in patients with Alzheimer's disease correlates to significant atrophy of these regions, detected by voxel-based morphometry, and to deterioration of specific cognitive functions.

Keywords: Alzheimer’s disease, functional MRI, voxel-based morphometry

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Authors: Owen Cegielski, Kristi Maida, Danny Morales, Sylvia L. Mendez

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This study explores the challenges and threats US secondary educators experience in incorporating culturally relevant education (CRE) practices in their classrooms. CRE is a social justice pedagogical practice used to connect student’s cultural references to academic skills and content, to promote critical reflection, to facilitate cultural competence, and to critique discourses of power and oppression. Empirical evidence on CRE demonstrates positive student educational outcomes in terms of achievement, engagement, and motivation. Additionally, due to the direct focus on uplifting diverse cultures through the curriculum, students experience greater feelings of belonging, increased interest in the subject matter, and stronger racial/ethnic identities. When these teaching practices are in place, educators develop deeper relationships with their students and appreciate the multitude of gifts they (and their families) bring to the classroom environment. Yet, educators regularly report being unprepared to incorporate CRE in their daily teaching practice and identify substantive gaps in their knowledge and skills in this area. Often, they were not exposed to CRE in their educator preparation program, nor do they receive adequate support through school- or district-wide professional development programming. Through a descriptive phenomenological research design, 20 interviews were conducted with a diverse set of secondary school educators to explore the challenges and threats they experience in incorporating CRE practices in their classrooms. The guiding research question for this study is: What are the challenges and threats US secondary educators face when seeking to incorporate CRE practices in their classrooms? Interviews were grounded by the theory of challenge and threat states, which highlights the ways in which challenges and threats are appraised and how resources factor into emotional valence and perception, as well as the potential to meet the task at hand. Descriptive phenomenological data analysis strategies were utilized to develop an essential structure of the educators’ views of challenges and threats in regard to incorporating CRE practices in their secondary classrooms. The attitude of the phenomenological reduction method was adopted, and the data were analyzed through five steps: sense of the whole, meaning units, transformation, structure, and essential structure. The essential structure that emerged was while secondary educators display genuine interest in learning how to successfully incorporate CRE practices, they perceive it to be a challenge (and not a threat) due to lack of exposure which diminishes educator capacity, comfort, and confidence in employing CRE practices. These findings reveal the value of attending to emotional valence and perception of CRE in promoting this social justice pedagogical practice. Findings also reveal the importance of appropriately resourcing educators with CRE support to ensure they develop and utilize this practice.

Keywords: culturally relevant education, descriptive phenomenology, social justice practice, US secondary education

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Authors: Leyla Sattarzadeh, Maghsoud Peeri, Mohammadali Azarbaijani, Hasan Matin Homaee

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Inflammation by various mechanisms may cause atherosclerosis. Systemic circulating inflammatory markers such as C-reactive protein (CRP), pro-inflammatory cytokines such as Interleukin-6 (IL-6) and adhesion molecules like Intracellular Adhesion Molecule-1 (ICAM-1) are the predictors of cardiovascular diseases. Regarding the conflicting results about the effect of resistance exercise training on these inflammatory markers, the present study aimed to examine the effect of eight week different patterns of resistance exercise training on CRP, IL-6 and ICAM-1 levels in healthy untrained women. 40 volunteered and healthy untrained female university students (aged: 21+ 3 yr., Body Mass Index: 21.5+ 3.5 kg/m2) were selected purposefully and divided into three groups. At the end of training protocol and after subjects drop during the protocol in upper body exercise training (n=11), lower body (n=12) completed the eight week of training period although the control group (n=7) did anything. Blood samples gathered pre and post experimental period and CRP, IL-6 and ICAM-1 levels were evaluated using special laboratory kits, then the difference of pre and post values of each indices analyzed using one way Analysis of Variance (α < 0.05). The results of one way ANOVA for difference of pre and post values of CRP and ICAM-1 showed no significant changes due to the exercise training. But there were significant differences between groups about IL-6. Tukey post- hoc test indicated that there is significant difference between the differences of pre and post values of IL-6 between lower body exercise training group and control group, and eight weeks of lower body exercise training lead to significant changes in IL-6 values. There were no changes in anthropometric indices. The findings show that the different patterns of upper and lower body exercise training by involving the different amount of muscles altered the IL-6 values in lower body exercise training group probably because of engaging the bigger amount of muscles, but showed any significant changes about CRP and ICAM-1 probably due to intensity and duration of exercise or the lower levels of these markers at baseline of healthy people.

Keywords: C-reactive protein, interleukin-6, intracellular adhesion molecule-1, resistance training

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Authors: N. A. P. Camaforte, P. M. P. Vareda, L. L. Saldanha, A. L. Dokkedal, J. M. Rezende-Neto, M. R. Senger, F. P. Silva-Jr, J. R. Bosqueiro

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Diabetes mellitus is a disease characterized by deficiency of insulin secretion and/or action which results in hyperglycemia. Nowadays, acarbose is a medicine used by diabetic people to inhibit alpha-glucosidases leading to the decreasing of post-feeding glycaemia, but with low effectiveness and many side effects. Medicinal plants have been used for the treatment of many diseases including diabetes and their action occurs through the modulation of insulin-depending processes, pancreas regeneration or inhibiting glucose absorption by the intestine. Previous studies in our laboratory showed that the treatment using two crude extracts of plants from Brazilian cerrado was able to decrease fasting blood glucose and improve glucose tolerance in streptozotocin-diabetic mice. Because of this and the importance of the search for new alternatives to decrease the hyperglycemia, we decided to evaluate the inhibitory action of two plants from Brazilian cerrado - B.H. and Myrcia bella. The enzymatic assay was performed in 50 µL of final volume using pancreatic α-amylase and maltase together with theirs commercial substrates. The inhibition potency (IC50) was determined by the incubation of eight different concentrations of both extracts and the enzymes for 5 minutes at 37ºC. After, the substrate was added to start the reaction. Glucosidases assay was evaluated measuring the quantity of p-nitrophenol in 405 nmin 384 wells automatic reader. The in vitro assay with the extracts of B.H. and M. bella showed an IC50 of 28,04µg/mL and 16,93 µg/mL for α-amilase, and 43,01µg/mL and 17 µg/mL for maltase, respectively. M. bella extract showed a higher inhibitory activity for those enzymes than B.H. extract. The crude extracts tested showed a higher inhibition rate to α-amylase, but were less effective against maltase in comparison to acarbose (IC50 36µg/mL and 9 µg/mL, respectively). In conclusion, the crude extract of B.H. and M. bella showed a potent inhibitory effect against α-amylase and showed promising results to the possible development of new medicines to treat diabetes with less or even without side effects.

Keywords: alfa-glucosidases, diabetes mellitus, glycaemia, medicinal plants

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Authors: Lokvendra S. Budania, Yogesh K Gaude, Vamsidhar Chamala

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Introduction: Congenitally corrected transposition of great arteries (CCTGA) is a complex congenital heart disease where there are both atrioventricular and ventriculoarterial discordances, usually accompanied by other cardiovascular malformations. Case Report: A 24-year-old primigravida known case of CCTGA at 37 weeks of gestation was referred to our hospital for safe delivery. Her electrocardiogram showed HR-40/pm, echocardiography showed Ejection Fraction of 65% and CCTGA. Temporary pacemaker was inserted by cardiologist in catheterization laboratory, before giving trial of labour in view of complete heart block. She was planned for normal delivery, but emergency Caesarean section was planned due to non-reassuring foetal Cardiotocography Pre-op vitals showed PR-50 bpm with temporary pacemaker, Blood pressure-110/70 mmHg, SpO2-99% on room air. Nil per oral was inadequate. Patency of two peripheral IV cannula checked and left radial arterial line secured. Epidural Anaesthesia was planned, and catheter was placed at L2-L3. Test dose was given, Anaesthesia was provided with 5ml + 5ml of 2% Lignocaine with 25 mcg Fentanyl and further 2.5Ml of 0.5% Bupivacaine was given to achieve a sensory level of T6. Cesarean section was performed and baby was delivered. Cautery was avoided during this procedure. IV Oxytocin (15U) was added to 500 mL of ringer’s lactate. Hypotension was treated with phenylephrine boluses. Patient was shifted to post-operative care unit and later to high dependency unit for monitoring. Post op vitals remained stable. Temporary pacemaker was removed after 24 hours of surgery. Her post-operative period was uneventful and discharged from hospital. Conclusion: Rare congenital cardiac disorders require detail knowledge of pathophysiology and associated comorbidities with the disease. Meticulously planned and carefully titrated neuraxial techniques will be beneficial for such cases.

Keywords: congenitally corrected transposition of great arteries, complete heart block, emergency LSCS, epidural anaesthesia

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Authors: Ali D. Omur, Betul Apaydin Yildirim, Yavuz S. Saglam, Selim Comakli, Mustafa Ozkaraca

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Twenty-eight male Sprague Dawley rats (aged 3 months) were used in the study. The animals were given feed and water as ad libitum. Sprague Dawley rats were randomly divided into 4 groups as 7 rats in each group. Aflatoxin B1 (7.5 μg/200 g), resveratrol (60 mg/kg) was administered to rats in groups other than the control group. At the end of the 16th day, blood, semen and tissue specimens were taken by decapitation under ether anesthesia. The effects of aflatoxin B1 and resveratrol on spermatological, pathological and biochemical parameters were determined in rats. When we evaluate the spermatological parameters, it is understood that resveratrol has a statistically significant difference in terms of sperm motility and viability (membrane integrity) compared to the control group and aflatoxin B1 administration groups, indicating a protective effect on spermatological parameters (groups: control, resveratrol, aflatoxin B1 and Afb1 + res; respectively, values of motility: 71,42 ± 0,52b, 72,85 ± 1, 48c , 60,71 ± 1,30a, 57,14 ± 2, 40a; values of viability: 63,85 ± 1,33b, 70,42 ± 2,61c, 55,00 ± 1,54a, 56,57 ± 0,89a. In terms of pathological parameters -histopathological examination- in the control and resveratrol groups, seminiferous tubules were observed to be in normal structure. In the group treated with aflatoxin, the regular structure of the spermatogenic cells deteriorated, and the seminiferous tubules became necrotic and degenerative. In the group treated with Afb1 + res, the decreasing of necrotic and degenerative changes were determined compared with in the group treated with aflatoxin. As immunohistochemical examination, cleaved caspase 3 expression was found to be very low in the control and resveratrol groups. Cleaved caspase 3 expression was severely exacerbated in seminiferous tubules in aflatoxin group but cleaved caspase 3 expression level decreased in Afb1 + res. In the biochemical direction, resveratrol has been shown to inhibit the adverse effects of aflatoxin on antioxidant levels (GSH-mmol/L, CAT-kU/L, GPx-U/mL, SOD-EU/mL) and to show a protective effect. For this purpose, the use of resveratrol with antioxidant activity was investigated in preventing or ameliorating damage to aflatoxin B1. It has been concluded that resveratrol effectively prevents the aflatoxin-induced testicular damage and lipid peroxidation. It has also been shown that resveratrol has protective effects on sperm motility and viability.

Keywords: Aflatoxin B1, rat, resveratrol, sperm

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Authors: Nadia Naseem, Farwa Batool, Nasir Mehmood, AbdulHannan Nagi

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Background: The incidence and mortality of breast cancer vary significantly in geographically distinct populations. In Pakistan, breast cancer has shown an increase in incidence in young females and is characterized by more aggressive behavior. The tumor suppressor TP53 gene is a crucial genetic factor that plays a significant role in breast carcinogenesis. This study investigated the TP53 mutations in molecular subtypes of both nodes negative and positive breast cancer in young Pakistani patients. Material and Methods: p53, Estrogen Receptor (ER), Progesterone Receptor (PR), Her-2 neu and Ki 67 expressions were analyzed immunohistochemically in a series of 75 node negative (A) and 75 node positive (B) young (aged: 19-40 years) breast cancer patients diagnosed between 2014 to 2017 at two leading hospitals of Punjab, Pakistan. Tumor tissue specimens and peripheral blood samples were examined for TP53 mutations by direct sequencing of the gene (exons 4-9). The relation of TP53 mutations to these markers and clinicopathological data was investigated. Results: Mean age of the patients was 32.4 + 9.1 SD. Invasive breast carcinoma was the most frequent histological variant (A=92%, B=94.6%). Grade 3 carcinoma was the commonest grade (A=72%, B=81.3%). Triple negative cases (ER-, PR-, Her-2) formed most of the molecular subtypes (A=44%, B=50.6%). A total of 17.2% (A: 6.6%, B: 10.6%) patients showed TP53 mutations. Mutations were significantly more frequent in triple negative cases (A: 74.8%, B: 62.2%) compared to HER2-positive patients (P < 0.0001). In the multivariate analysis of the whole patient group, the independent prognosticator were triple negative cases (P=0.021), TP53 overexpression by IHC (P=0.001) and advanced-stage disease (P=0.007). No statistically significant correlation between TP53 mutations and clinicopathological parameters was found (P < 0.05). Conclusions: It is concluded that TP53 mutations are infrequently present in breast carcinoma of young Pakistani population and there was no significant correlation between p53 mutation and early onset disease. Immunohistochemically detected TP53 expression in our resource-constrained to set up can be beneficial in predicting mutations at the younger age in our population.

Keywords: immunohistochemistry (IHC), invasive breast carcinoma (IBC), Pakistan, TP53

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Authors: Turki J. Alharbi, Jencia Wong, Dennis Yue, Tania P. Markovic, Julie Hetherington, Ted Wu, Belinda Brooks, Radhika Seimon, Alice Gibson, Stephanie L. Silviera, Amanda Sainsbury, Tanya J. Little

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Fasting has been practiced for centuries and is incorporated into the practices of different religions including Islam, whose followers intermittently fast throughout the month of Ramadan. Thus, Ramadan presents a unique model of prolonged intermittent fasting (IF). Despite a growing body of evidence for a cardio-metabolic and endocrine benefit of IF, detailed studies of the effects of IF on these indices in type 2 diabetes are scarce. We studied 5 subjects with type 2 diabetes (T2DM) and 7 healthy controls (C) at baseline (pre), and in the last week of Ramadan (post). Fasting circulating levels of glucose, HbA1c and lipids, as well as body composition (with DXA) and resting energy expenditure (REE) were measured. Plasma gut hormone levels and appetite responses to a mixed meal were also studied. Data are means±SEM. Ramadan decreased total fat mass (-907±92 g, p=0.001) and trunk fat (-778±190 g, p=0.014) in T2DM but not in controls, without any reductions in lean mass or REE. There was a trend towards a decline in plasma FFA in both groups. Ramadan had no effect on body weight, glycemia, blood pressure, or plasma lipids in either group. In T2DM only, the area under the curve for post-meal plasma ghrelin concentrations increased after Ramadan (pre:6632±1737 vs. post:9025±2518 pg/ml.min-1, p=0.045). Despite this increase in orexigenic ghrelin, subjective appetite scores were not altered by Ramadan. Meal-induced plasma concentrations of the satiety hormone pancreatic polypeptide did not change during Ramadan, but were higher in T2DM compared to controls (post: C: 23486±6677 vs. T2DM: 62193±6880 pg/ml.min-1, p=0.003. In conclusion, Ramadan, as a model for IF appears to have more favourable effects on body composition in T2DM, without adverse effects on metabolic control or subjective appetite. These data suggest that IF may be particularly beneficial in T2DM as a nutritional intervention. Larger studies are warranted.

Keywords: type 2 diabetes, obesity, intermittent fasting, appetite regulating hormones

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Authors: Ammar Asma, Bouafia Nabiha, Ghammam Rim, Ezzi Olfa, Ben Cheikh Asma, Mahjoub Mohamed, Helali Radhia, Sma Nesrine, Chouchène Imed, Boussarsar Hamadi, Njah Mansour

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Background: Central venous catheter associated infections (CVC-AI) are among the serious hospital-acquired infections. The aims of this study are to determine the incidence of CVC-AI, and their risk factors among patients followed in a Tunisian medical intensive care unit (ICU). Materials / Methods: A prospective cohort study conducted between September 15th, 2015 and November 15th, 2016 in an 8-bed medical ICU including all patients admitted for more than 48h. CVC-AI were defined according to CDC of ATLANTA criteria. The enrollment was based on clinical and laboratory diagnosis of CVC-AI. For all subjects, age, sex, underlying diseases, SAPS II score, ICU length of stay, exposure to CVC (number of CVC placed, site of insertion and duration catheterization) were recorded. Risk factors were analyzed by conditional stepwise logistic regression. The p-value of < 0.05 was considered significant. Results: Among 192 eligible patients, 144 patients (75%) had a central venous catheter. Twenty-eight patients (19.4%) had developed CVC-AI with density rate incidence 20.02/1000 CVC-days. Among these infections, 60.7% (n=17) were systemic CVC-AI (with negative blood culture), and 35.7% (n=10) were bloodstream CVC-AI. The mean SAPS II of patients with CVC-AI was 32.76 14.48; their mean Charlson index was 1.77 1.55, their mean duration of catheterization was 15.46 10.81 days and the mean duration of one central line was 5.8+/-3.72 days. Gram-negative bacteria was determined in 53.5 % of CVC-AI (n= 15) dominated by multi-drug resistant Acinetobacter baumani (n=7). Staphylococci were isolated in 3 CVC-AI. Fourteen (50%) patients with CVC-AI died. Univariate analysis identified men (p=0.034), the referral from another hospital department (p=0.03), tobacco (p=0.006), duration of sedation (p=0.003) and the duration of catheterization (p=0), as possible risk factors of CVC-AI. Multivariate analysis showed that independent factors of CVC-AI were, male sex; OR= 5.73, IC 95% [2; 16.46], p=0.001, Ramsay score; OR= 1.57, IC 95% [1.036; 2.38], p=0.033, and duration of catheterization; OR=1.093, IC 95% [1.035; 1.15], p=0.001. Conclusion: In a monocenter cohort, CVC-AI had a high density and is associated with poor outcome. Identifying the risk factors is necessary to find solutions for this major health problem.

Keywords: central venous catheter associated infection, intensive care unit, prospective cohort studies, risk factors

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Authors: Maria Dora Horvath, Norbert Buzas, Zsanett Tesch

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Diabetes management poses many unique challenges for children and their parents. The use of a diabetes management device should not be one of these challenges as the purpose of these devices is to make the management more convenient. The objective of our study was to examine how demographical, psychological and diabetes-related factors determine the choices parents make regarding their child’s diabetes management technologies and how they perceive advanced devices. We conducted the study using an online questionnaire with 318 parents (mostly mothers). The questions of the survey were about demographical, diabetes-related and psychological factors (diabetes management problems, diabetes management competence). In addition, we asked the parents opinions about advanced diabetes management devices. We expanded our data with semi-structured in-depth interviews. 61 % of the participants Self-Monitored Blood Glucose (SMBG), and 39 % used a Continuous Glucose Monitoring System (CGM). Considering insulin administration, 58 % used Multiple Daily Insulin Injections (MDII) and 42 % used Continuous Subcutaneous Insulin Infusion (CSII). Parents who used diverse combinations of diabetes management devices showed significant differences in age (parents’ and child’s), the monthly cost of diabetes, the duration of diabetes, the highest level of education and average monthly household income. CGM users perceived diabetes management problems significantly more severe than SMBG users and CSII users felt significantly more competent in diabetes management than MDII users. Avoiding CGM use due to lack of financial resources was determined by diagnosis duration. While avoiding its use by the cause of the child rejecting, it was determined by the child’s age and diabetes competence. Using MDII instead of CSII because of the child’s rejection was determined by the monthly cost of diabetes and child’s age. We conducted a complex empirical study in which we examined perceptions and experiences of advanced and less advanced diabetes management technologies comprehensively. Our study highlights the factors that fundamentally influence parents’ motivations and choices about diabetes management technologies. These results could contribute to developing diabetes management technologies more suitable for children living with type 1 diabetes and their parents.

Keywords: advanced diabetes management technologies, children living with type 1 diabetes, diabetes management, motivation, parents

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Authors: Hollie Ashworth, Sarah Heath, Nick Bryan, Liam Abrahamsen, Simon Kellet

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Sellafield has a number of fuel storage ponds, some of which have been open to the air for a number of decades. This has caused corrosion of the fuel resulting in a release of some activity into solution, reduced water clarity, and accumulation of sludge at the bottom of the pond consisting of brucite (Mg(OH)2) and other uranium corrosion products. Both of these phases are also present as colloidal material. 90Sr and 137Cs are known to constitute a small volume of the radionuclides present in the pond, but a large fraction of the activity, thus they are most at risk of challenging effluent discharge limits. Organic molecules are known to be present also, due to the ponds being open to the air, with occasional algal blooms restricting visibility further. The contents of the pond need to be retrieved and safely stored, but dealing with such a complex, undefined inventory poses a unique challenge. This work aims to determine and understand the sorption-desorption interactions of 90Sr and 137Cs to brucite and uranium phases, with and without the presence of organic molecules from chemical degradation and bio-organisms. The influence of organics on these interactions has not been widely studied. Partitioning of these radionuclides and organic molecules has been determined through LSC, ICP-AES/MS, and UV-vis spectrophotometry coupled with ultrafiltration in both binary and ternary systems. Further detailed analysis into the surface and bonding environment of these components is being investigated through XAS techniques and PHREEQC modelling. Experiments were conducted in CO2-free or N2 atmosphere across a high pH range in order to best simulate conditions in the pond. Humic acid used in brucite systems demonstrated strong competition against 90Sr for the brucite surface regardless of the order of addition of components. Variance of pH did have a small effect, however this range (10.5-11.5) is close to the pHpzc of brucite, causing the surface to buffer the solution pH towards that value over the course of the experiment. Sorption of 90Sr to UO2 obeyed Ho’s rate equation and demonstrated a slow second-order reaction with respect to the sharing of valence electrons from the strontium atom, with the initial rate clearly dependent on pH, with the equilibrium concentration calculated at close to 100% sorption. There was no influence of humic acid seen when introduced to these systems. Sorption of 137Cs to UO3 was significant, with more than 95% sorbed in just over 24 hours. Again, humic acid showed no influence when introduced into this system. Both brucite and uranium based systems will be studied with the incorporation of cyanobacterial cultures harvested at different stages of growth. Investigation of these systems provides insight into, and understanding of, the effect of organics on radionuclide partitioning to brucite and uranium phases at high pH. The majority of sorption-desorption work for radionuclides has been conducted at neutral to acidic pH values, and mostly without organics. These studies are particularly important for the characterisation of legacy wastes at Sellafield, with a view to their safe retrieval and storage.

Keywords: caesium, legacy wastes, organics, sorption-desorption, strontium, uranium

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Authors: Ankit Kumar Yadav, Nagendra R. Velaga

Abstract:

Driving under the influence of alcohol impairs the driving performance and increases the crash risks worldwide. The present study investigated the effect of different Blood Alcohol Concentrations (BAC) on the accelerating and braking behaviour of drivers with the help of driving simulator experiments. Eighty-two licensed Indian drivers drove on the rural road environment designed in the driving simulator at BAC levels of 0.00%, 0.03%, 0.05%, and 0.08% respectively. Driving performance was analysed with the help of vehicle control performance indicators such as mean acceleration and mean brake pedal force of the participants. Preliminary analysis reported an increase in mean acceleration and mean brake pedal force with increasing BAC levels. Generalized linear mixed models were developed to quantify the effect of different alcohol levels and explanatory variables such as driver’s age, gender and other driver characteristic variables on the driving performance indicators. Alcohol use was reported as a significant factor affecting the accelerating and braking performance of the drivers. The acceleration model results indicated that mean acceleration of the drivers increased by 0.013 m/s², 0.026 m/s² and 0.027 m/s² for the BAC levels of 0.03%, 0.05% and 0.08% respectively. Results of the brake pedal force model reported that mean brake pedal force of the drivers increased by 1.09 N, 1.32 N and 1.44 N for the BAC levels of 0.03%, 0.05% and 0.08% respectively. Age was a significant factor in both the models where one year increase in drivers’ age resulted in 0.2% reduction in mean acceleration and 19% reduction in mean brake pedal force of the drivers. It shows that driving experience could compensate for the negative effects of alcohol to some extent while driving. Female drivers were found to accelerate slower and brake harder as compared to the male drivers which confirmed that female drivers are more conscious about their safety while driving. It was observed that drivers who were regular exercisers had better control on their accelerator pedal as compared to the non-regular exercisers during drunken driving. The findings of the present study revealed that drivers tend to be more aggressive and impulsive under the influence of alcohol which deteriorates their driving performance. Drunk driving state can be differentiated from sober driving state by observing the accelerating and braking behaviour of the drivers. The conclusions may provide reference in making countermeasures against drinking and driving and contribute to traffic safety.

Keywords: alcohol, acceleration, braking behaviour, driving simulator

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Authors: Sena Park

Abstract:

Background: The concurrent presentation with colorectal cancer in the inguinal hernia has been extremely rare. Due to its rarity, its presentation may lead to diagnostic and therapeutic dilemmas. We aim to review all the reported cases on colorectal cancer incarcerated in the inguinal hernia in the last 20 years, and discuss the operative approaches. Methods: We identified all case reports on colorectal cancer within inguinal hernia using PUBMED (2002-2022) and MEDLINE (2002-2022). The search strategy included the following keywords: colorectal cancer (title/abstract) AND inguinal hernia (title/abstract) OR incarceration (title/abstract). The search did not include letters, book chapters, systemic reviews, meta-analysis and editorials. Results: In the last 20 years, a total of 19 cases on colorectal cancer within the inguinal hernia were identified. The age of the patients ranged between 48 and 89. Majority of the patients were male (95%). Most commonly involved part of the large intestine was sigmoid colon (79%). Of all the cases, 79 percent of patients received open procedure and 21 percent had laparoscopic procedure. Discussion: Inguinal hernias are common with an incidence of approximately 1.7 percent. Colorectal cancer is the one of the leading causes of cancer-related mortality worldwide. However, their concurrent presentation has been extremely rare. In the last 20 years, 19 cases on concurrent presentation of colorectal cancer and inguinal hernia have been reported. Most patients who had open procedures had two incisions of groin incision and a midline laparotomy. There were 4 cases where the oncological resection was performed laparoscopically. The advantages of laparoscopic resection include reduced blood lost, reduced post-operative pain, reduced length of hospital stay and similar number of lymph nodes taken. From the review of the cases in the last 20 years, both open and laparoscopic approaches seemed to be safe and achieve adequate oncological resections. Conclusion: This is a brief overview of reported cases of colorectal cancer presenting with inguinal hernia concurrently. Due to its rarity, there are no current guidelines on operative approach in clinical practice. The experience in the last 20 years supports both open and laparoscopic approach.

Keywords: colorectal cancer, inguinal hernia, incarceration, operative approach

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Authors: Katie Emerson, Sara Perez-Ojalvo, Jim Komorowski, Danielle Greenberg

Abstract:

The purpose of this study was to evaluate arginase activity levels in response to combinations of an inositol-stabilized arginine silicate (ASI; Nitrosigine®), L-arginine, and Inositol. Arginine acts as a vasodilator that promotes increased blood flow resulting in enhanced delivery of oxygen and nutrients to the brain and other tissues. ASI alone has been shown to improve performance on cognitive tasks. Arginase, found in human serum, catalyzes the conversion of arginine to ornithine and urea, completing the last step in the urea cycle. Decreasing arginase levels maintains arginine and results in increased nitric oxide production. This study aimed to determine the most effective combination of ASI, L-arginine and inositol for minimizing arginase levels and therefore maximize ASI’s effect on cognition. Serum was taken from untreated healthy donors by separation from clotted factors. Arginase activity of serum in the presence or absence of test products was determined (QuantiChrom™, DARG-100, Bioassay Systems, Hayward CA). The remaining ultra-filtrated serum units were harvested and used as the source for the arginase enzyme. ASI alone or combined with varied levels of Inositol were tested as follows: ASI + inositol at 0.25 g, 0.5 g, 0.75 g, or 1.00 g. L-arginine was also tested as a positive control. All tests elicited changes in arginase activity demonstrating the efficacy of the method used. Adding L-arginine to serum from untreated subjects, with or without inositol only had a mild effect. Adding inositol at all levels reduced arginase activity. Adding 0.5 g to the standardized amount of ASI led to the lowest amount of arginase activity as compared to the 0.25g 0.75g or 1.00g doses of inositol or to L-arginine alone. The outcome of this study demonstrates an interaction of the pairing of inositol with ASI on the activity of the enzyme arginase. We found that neither the maximum nor minimum amount of inositol tested in this study led to maximal arginase inhibition. Since the inhibition of arginase activity is desirable for product formulations looking to maintain arginine levels, the most effective amount of inositol was deemed preferred. Subsequent studies suggest this moderate level of inositol in combination with ASI leads to cognitive improvements including reaction time, executive function, and concentration.

Keywords: arginine, inositol, arginase, cognitive benefits

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