Search results for: serum protein electrophoresis

Authors: Sairi Satari

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Introduction: Six-sigma is a metric that quantifies the performance of processes as a rate of Defects-Per-Million Opportunities. Sigma methodology can be applied in chemical pathology laboratory for evaluating process performance with evidence for process improvement in quality assurance program. In the laboratory, these methods have been used to improve the timeliness of troubleshooting, reduce the cost and frequency of quality control and minimize pre and post-analytical errors. Aim: The aim of this study is to evaluate the sigma values of the Cobas 8000 analyzer based on the minimum requirement of the specification. Methodology: Twenty-one analytes were chosen in this study. The analytes were alanine aminotransferase (ALT), albumin, alkaline phosphatase (ALP), Amylase, aspartate transaminase (AST), total bilirubin, calcium, chloride, cholesterol, HDL-cholesterol, creatinine, creatinine kinase, glucose, lactate dehydrogenase (LDH), magnesium, potassium, protein, sodium, triglyceride, uric acid and urea. Total error was obtained from Clinical Laboratory Improvement Amendments (CLIA). The Bias was calculated from end cycle report of Royal College of Pathologists of Australasia (RCPA) cycle from July to December 2016 and coefficient variation (CV) from six-month internal quality control (IQC). The sigma was calculated based on the formula :Sigma = (Total Error - Bias) / CV. The analytical performance was evaluated based on the sigma, sigma > 6 is world class, sigma > 5 is excellent, sigma > 4 is good and sigma < 4 is satisfactory and sigma < 3 is poor performance. Results: Based on the calculation, we found that, 96% are world class (ALT, albumin, ALP, amylase, AST, total bilirubin, cholesterol, HDL-cholesterol, creatinine, creatinine kinase, glucose, LDH, magnesium, potassium, triglyceride and uric acid. 14% are excellent (calcium, protein and urea), and 10% ( chloride and sodium) require more frequent IQC performed per day. Conclusion: Based on this study, we found that IQC should be performed frequently for only Chloride and Sodium to ensure accurate and reliable analysis for patient management.

Keywords: sigma matrics, analytical performance, total error, bias

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Authors: Zohreh Afsharmand, Sokhanguei Yahya

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A growing body of literature suggests that that low-grade systemic inflammation associated to obesity plays a key role in the pathogenic mechanism of several disorders. In this study, the effect of 6 weeks aerobic training on IL-6 and IL-1B as inflammatory cytokine were investigated in adult obese women. For this purpose, 26 sedentary adult obese women were divided into exercise and control groups (n=12). Pre and post training of mentioned cytokines were measured in two groups. Student’s t-tests for paired samples were performed to determine whether there were significant within-group changes in the outcomes. A p value less than 0.05 was considered statistically significant. There were no statistically significant differences between the exercise and control groups with regard to anthropometrical markers or inflammatory cytokines. Despite the significant decrease in all anthropometrical markers, no significant differences were found in serum IL-6 and IL-1B by aerobic training with compared to baseline. Our findings indicate that aerobic training intervention for a short time is not associated with the anti-inflammatory property in obese women.

Keywords: aerobic training, cytokine, inflammation, obesity

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Authors: Nizar Chaira

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Dates, main products of the oases, due to their therapeutic interests, are considered highly nutritious fruit. Several studies on the valuation biotechnology and technology of dates are made, and several products are already prepared. Isolation of the yeast Saccharomyces cerevisiae, naturally presents in a scrap of date, optimization of growth in the medium based on date syrup and production biomass can potentially expand the range of secondary products of dates. To this end, this paper tries to study the suitability for processing dates technology and biotechnology to use the date pulp as a carbon source for biological transformation. Two strains of Saccharomyces cerevisiae isolated from date syrup (S1, S2) and a commercial strain have used for this study. After optimization of culture conditions, production in a fermenter on two different media (date syrup and beet molasses) was performed. This is followed by studying the kinetics of growth, protein production and consumption of sugars in crops strain 1, 2 and the commercial strain and on both media. The results obtained showed that a concentration of 2% sugar, 2.5 g/l yeast extract, pH 4.5 and a temperature between 25 and 35°C are the optimal conditions for cultivation in a bioreactor. The exponential phase of the specific growth rate of a strain on both media showed that it is about 0.3625 h-1 for the production of a medium based on date syrup and 0.3521 h-1 on beet molasses with a generation time equal to 1.912 h and on the medium based on date syrup, yeast consumes preferentially the reducing sugars. For the production of protein, we showed that this latter presents an exponential phase when the medium starts to run out of reducing sugars. For strain 2, the specific growth rate is about 0.261h-1 for the production on a medium based on date syrup and 0207 h-1 on beet molasses and the base medium syrup date of the yeast consumes preferentially reducing sugars. For the invertase and other metabolits, these increases rapidly after exhaustion of reducing sugars. The comparison of productivity between the three strains on the medium based on date syrup showed that the maximum value is obtained with the second strain: p = 1072 g/l/h as it is about of 0923 g/l/h for strain 1 and 0644 g/l/h for the commercial strain. Thus, isolates of date syrup are more competitive than the commercial strain and can give the same performance in a shorter time with energy gain.

Keywords: date palm, fermentation, molasses, Saccharomyces, syrup

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Authors: Bright Chukwunwike Uzuegbunam, Wojciech Paslawski, Hans Agren, Christer Halldin, Wolfgang Weber, Markus Luster, Thomas Arzberger, Behrooz Hooshyar Yousefi

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There is a need to develop a PET tracer that will enable to diagnosis and track the progression of Alpha-synucleinopathies (Parkinson’s disease [PD], dementia with Lewy bodies [DLB], multiple system atrophy [MSA]) in living subjects over time. Alpha-synuclein aggregates (a-syn), which are present in all the stages of disease progression, for instance, in PD, are a suitable target for in vivo PET imaging. For this reason, we have developed some promising a-syn tracers based on a disarylbisthiazole (DABTA) scaffold. The precursors are synthesized via a modified Hantzsch thiazole synthesis. The precursors were then radiolabeled via one- or two-step radiofluorination methods. The ligands were initially screened using a combination of molecular dynamics and quantum/molecular mechanics approaches in order to calculate the binding affinity to a-syn (in silico binding experiments). Experimental in vitro binding assays were also performed. The ligands were further screened in other experiments such as log D, in vitro plasma protein binding & plasma stability, biodistribution & brain metabolite analyses in healthy mice. Radiochemical yields were up to 30% - 72% in some cases. Molecular docking revealed possible binding sites in a-syn and also the free energy of binding to those sites (-28.9 - -66.9 kcal/mol), which correlated to the high binding affinity of the DABTAs to a-syn (Ki as low as 0.5 nM) and selectivity (> 100-fold) over Aβ and tau, which usually co-exist with a-synin some pathologies. The log D values range from 2.88 - 2.34, which correlated with free-protein fraction of 0.28% - 0.5%. Biodistribution experiments revealed that the tracers are taken up (5.6 %ID/g - 7.3 %ID/g) in the brain at 5 min (post-injection) p.i., and cleared out (values as low as 0.39 %ID/g were obtained at 120 min p.i. Analyses of the mice brain 20 min p.i. Revealed almost no radiometabolites in the brain in most cases. It can be concluded that in silico study presents a new venue for the rational development of radioligands with suitable features. The results obtained so far are promising and encourage us to further validate the DABTAs in autoradiography, immunohistochemistry, and in vivo imaging in non-human primates and humans.

Keywords: alpha-synuclein aggregates, alpha-synucleinopathies, PET imaging, tracer development

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Authors: Danutė Jablonskytė-Raščė, Vidas Damanauskas

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The wheat (Triticum) genus is divided into many species, of which only two are widely distributed in the world - common wheat (Triticum aestivum L.) and durum wheat (Triticum durum Desf.). Common (soft) wheat is the most common type of wheat in the world and the most suitable for the harsh climate of Lithuania, but the grains have lower protein content and poorer nutritional properties. Durum wheat is characterized by a high protein content of the grain, but it is a crop of warmer climates grown in southern countries, Italy, Spain, the United States, Egypt, etc. Today's important issue is food, its resources and quality. The research focuses on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the warming climate conditions. Climatic conditions change the distribution of fungi and their hosts. Plants that have grown in our climate for many years have adapted to the use of fungicides, so the aim is to study cereal varieties grown in warmer climates and compare them with our country's varieties, studying their nutritional value and the spread of fungal diseases. The field experiments of different varieties of wheat were conducted at Joniškėlis Experimental Station of the Lithuanian Research Centre for Agriculture and Forestry in 2023. The soil of the experimental site was Endocalcari-Endohypogleyic Cambisol (CMg-n-w-can). The research was designed to identify the resistance to leaf diseases and the nutritional value of various wheat varieties. This research aims to focus on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the conditions of the warming climate. The study found that hot and humid summer weather led to the spread of foliar diseases in wheat. Tan spot (Pyrenophora tritici-repentis) is mostly spread in wheat crops. This disease had an average prevalence of 86.90%. The wheat crop was sparse, so this year was unfavorable for the spread of powdery mildew (Blumeria graminis). Dry weather prevailed during the period of flowering of cereals, which prevented the spread of ear diseases. Examining the qualitative indicators of grain, it was found that durum wheat had the best parameters.

Keywords: varieties, wheat, leaf disease, grain quality

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Authors: Salina Yahya Saddick

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Ovarian cancer remains the most lethal gynecological malignancy due to the lack of highly sensitive and specific screening tools for detection of early-stage disease. The OSE provides the progenitor cells for 90% of human ovarian cancers. Recent morphologic, immunohistochemical and molecular genetic studies have led to the development of a new paradigm for the pathogenesis and origin of epithelial ovarian cancer (EOC) based on a ualistic model of carcinogenesis that divides EOC into two broad categories designated Types I and II which are characterized by specific mutations, including KRAS, BRAF, ERBB2, CTNNB1, PTEN PIK3CA, ARID1A, and PPPR1A, which target specific cell signaling pathways. Type 1 tumors rarely harbor TP53. type I tumors are relatively genetically stable and typically display a variety of somatic sequence mutations that include KRAS, BRAF, PTEN, PIK3CA CTNNB1 (the gene encoding beta catenin), ARID1A and PPP2R1A but very rarely TP53 . The cancer stem cell (CSC) hypothesis postulates that the tumorigenic potential of CSCs is confined to a very small subset of tumor cells and is defined by their ability to self-renew and differentiate leading to the formation of a tumor mass. Potential protein biomarker miRNA, are promising biomarkers as they are remarkably stable to allow isolation and analysis from tissues and from blood in which they can be found as free circulating nucleic acids and in mononuclear cells. Recently, genomic anaylsis have identified biomarkers and potential therapeutic targets for ovarian cancer namely, FGF18 which plays an active role in controlling migration, invasion, and tumorigenicity of ovarian cancer cells through NF-κB activation, which increased the production of oncogenic cytokines and chemokines. This review summarizes update information on epithelial ovarian cancers and point out to the most recent ongoing research.

Keywords: epithelial ovarian cancers, somatic sequence mutations, cancer stem cell (CSC), potential protein, biomarker, genomic analysis, FGF18 biomarker

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Authors: Mei Chen, Yingping Hou, Michelle Hao, Soheil Aghamohammadzadeh, Esteban Terzo, Roger Clark, Vijay Modur

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Background: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a genetic skin condition characterized by skin tearing and unremitting blistering upon minimal trauma. Repeated blistering, fibrosis, and scarring lead to aggressive squamous cell carcinoma later in life. RDEB is caused by mutations in the COL7A1 gene encoding collagen type VII (C7), the major component of anchoring fibrils mediating epidermis-dermis adherence. Nonsense mutations in the COL7A1 gene of a subset of RDEB patients leads to premature termination codons (PTC). Similarly, most Junctional Epidermolysis Bullosa (JEB) cases are caused by nonsense mutations in the LAMB3 gene encoding the β3 subunit of laminin 332. Currently, there is an unmet need for the treatment of RDEB and JEB. Zikani Therapeutics has discovered an array of macrocyclic compounds with ring structures similar to macrolide antibiotics that can facilitate readthrough activity of nonsense mutations in the COL7A1 and LAMB3 genes by acting as Ribosome Modulating Agents (RMAs). The medicinal chemistry synthetic advancements of these macrocyclic compounds have allowed targeting the human ribosome while preserving the structural elements responsible for the safety and pharmacokinetic profile of clinically used macrolide antibiotics. Methods: C7 expression was used as a measure of readthrough activity by immunoblot assays in two primary human fibroblasts from RDEB patients (R578X/R578X and R163X/R1683X-COL7A1). Similarly, immunoblot assays in C325X/c.629-12T > A-LAMB3 keratinocytes were used to measure readthrough activity for JEB. The relative readthrough activity of each compound was measured relative to Gentamicin. An imaging-based fibroblast migration assay was used as an assessment of C7 functionality in RDEB-fibroblasts over 16-20 hrs. The incubation period for the above experiments was 48 hrs for RDEB fibroblasts and 72 hours for JEB keratinocytes. Results: 9 RMAs demonstrated increased protein expression in both patient RDEB fibroblasts. The highest readthrough activity at tested concentrations without cytotoxicities increased protein expression up to 179% of Gentamicin (400 µg/ml), with favored readthrough activity in R163X/R1683X-COL7A1 fibroblasts. Concurrent with protein expression, fibroblast hypermotility phenotype observed in RDEB was rescued by reducing motility by ~35% to WT levels (the same level as 690 µM Gentamicin treated cells). Laminin β3 expression was also shown to be increased by 6 RMAs in keratinocytes to 33-83% of (400 µg/ml) Gentamicin. Conclusions: To date, 9 RMAs have been identified that enhance the expression of functional C7 in a mutation-dependent manner in two different RDEB patient fibroblast backgrounds (R578X/R578X and R163X/R1683X-COL7A1). A further 6 RMAs have been identified that enhance the readthrough of C325X-LAMB3 in JEB patient keratinocytes. Based on the clinical trial conducted by us with topical gentamycin in 2017, Zikani’s RMAs achieve clinically significant levels of read-through for the treatment of recessive dystrophic and Junctional Epidermolysis Bullosa.

Keywords: epidermolysis bullosa, nonsense mutation, readthrough, ribosome modulation

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Authors: Dar-Bin Shieh, Gwo-Bin Lee, Chen-Ming Chang, Chen Sheng Yeh, Chih-Chia Huang, Tsung-Ju Li

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Mitochondrial defects have a significant impact in many human diseases and aging associated phenotypes. The pathogenic mitochondrial DNA (mtDNA) mutations are diverse and usually present as heteroplasmic. mtDNA 4977bps deletion is one of the common mtDNA defects, and the ratio of mutated versus normal copy is significantly associated with clinical symptoms thus their quantitative detection has become an important unmet needs for advanced disease diagnosis and therapeutic guidelines. This study revealed a Micro-electro-mechanical-system (MEMS) enabled automatic microfluidic chip that only required minimal sample. The system integrated multiple laboratory operation steps into a Lab-on-a-Chip for high-sensitive and prompt measurement. The entire process including magnetic nanoparticle based mtDNA extraction in chip, mutation selective photonic DNA cleavage, and nanoparticle accelerated photonic quantitative polymerase chain reaction (qPCR). All subsystems were packed inside a miniature three-dimensional micro structured system and operated in an automatic manner. Integration of magnetic beads with microfluidic transportation could promptly extract and enrich the specific mtDNA. The near infrared responsive magnetic nanoparticles enabled micro-PCR to be operated by pulse-width-modulation controlled laser pulsing to amplify the desired mtDNA while quantified by fluorescence intensity captured by a complementary metal oxide system array detector. The proportions of pathogenic mtDNA in total DNA were thus obtained. Micro capillary electrophoresis module was used to analyze the amplicone products. In conclusion, this study demonstrated a new magnetophotonic based qPCR MEMS system that successfully detects and quantify specific disease related DNA mutations thus provides a promising future for rapid diagnosis of mitochondria diseases.

Keywords: mitochondrial DNA, micro-electro-mechanical-system, magnetophotonics, PCR

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Authors: Ogechi Nzeagwu, Assumpta Osuagwu, Charlse Nkwoala

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Malnutrition in infants due to poverty, poor feeding practices, and high cost of commercial complementary foods among others is a concern in developing countries. The study evaluated the proximate, vitamin and mineral compositions, antinutrients and functional properties, biochemical, haematological and sensory evaluation of complementary food made from sorghum, groundnut, crayfish and paw-paw flour blends using standard procedures. The blends were formulated on protein requirement of infants (18 g/day) using Nutrisurvey linear programming software in ratio of sorghum(S), groundnut(G), crayfish(C) and pawpaw(P) flours as 50:25:10:15(SGCP1), 60:20:10:10 (SGCP2), 60:15:15:10 (SGCP3) and 60:10:20:10 (SGCP4). Plain-pap (fermented maize flour)(TCF) and cerelac (commercial complementary food) served as basal and control diets. Thirty weanling male albino rats aged 28-35 days weighing 33-60 g were purchased and used for the study. The rats after acclimatization were fed with gruel produced with the experimental diets and the control with water ad libitum daily for 35days. Effect of the blends on lipid profile, blood glucose, haematological (RBC, HB, PCV, MCV), liver and kidney function and weight gain of the rats were assessed. Acceptability of the gruel was conducted at the end of rat feeding on forty mothers of infants’ ≥ 6 months who gave their informed consent to participate using a 9 point hedonic scale. Data was analyzed for means and standard deviation, analysis of variance and means were separated using Duncan multiple range test and significance judged at 0.05, all using SPSS version 22.0. The results indicated that crude protein, fibre, ash and carbohydrate of the formulated diets were either comparable or higher than values in cerelac. The formulated diets (SGCP1- SGCP4) were significantly (P>0.05) higher in vitamin A and thiamin compared to cerelac. The iron content of the formulated diets SGCP1- SGCP4 (4.23-6.36 mg/100) were within the recommended iron intake of infants (0.55 mg/day). Phytate (1.56-2.55 mg/100g) and oxalate (0.23-0.35 mg/100g) contents of the formulated diets were within the permissible limits of 0-5%. In functional properties, bulk density, swelling index, % dispersibility and water absorption capacity significantly (P<0.05) increased and compared favourably with cerelac. The essential amino acids of the formulated blends were within the amino acid profile of the FAO/WHO/UNU reference protein for children 0.5 -2 years of age. Urea concentration of rats fed with SGCP1-SGCP4 (19.48 mmol/L),(23.76 mmol/L),(24.07 mmol/L),(23.65 mmol/L) respectively was significantly higher than that of rat fed cerelac (16.98 mmol/L); however, plain pap had the least value (9.15 mmol/L). Rats fed with SGCP1-SGCP4 (116 mg/dl), (119 mg/dl), (115 mg/dl), (117 mg/dl) respectively had significantly higher glucose levels those fed with cerelac (108 mg/dl). Liver function parameters (AST, ALP and ALT), lipid profile (triglyceride, HDL, LDL, VLDL) and hematological parameters of rats fed with formulated diets were within normal range. Rats fed SGCP1 gained more weight (90.45 g) than other rats fed with SGCP2-SGCP4 (71.65 g, 79.76 g, 75.68 g), TCF (20.13 g) and cerelac (59.06 g). In all the sensory attributes, the control was preferred with respect to the formulated diets. The formulated diets were generally adequate and may likely have potentials to meet nutrient requirements of infants as complementary food.

Keywords: biochemical, chemical evaluation, complementary food, quadrimix

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Authors: A. K. Faizah, G. Vadamalai, S. K. Balasundram, W. L. Lim

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Pineapple (Ananas comosus) is a common crop in tropical and subtropical areas of the world. Malaysia once ranked as one of the top 3 pineapple producers in the world in the 60's and early 70's, after Hawaii and Brazil. Moreover, government’s recognition of the pineapple crop as one of priority commodities to be developed for the domestics and international markets in the National Agriculture Policy. However, pineapple industry in Malaysia still faces numerous challenges, one of which is the management of disease and pest. Red tip disease on pineapple was first recognized about 20 years ago in a commercial pineapple stand located in Simpang Renggam, Johor, Peninsular Malaysia. Since its discovery, there has been no confirmation on its causal agent of this disease. The epidemiology of red tip disease is still not fully understood. Nevertheless, the disease symptoms and the spread within the field seem to point toward viral infection. Bioassay test on nucleic acid extracted from the red tip-affected pineapple was done on Nicotiana tabacum cv. Coker by rubbing the extracted sap. Localised lesions were observed 3 weeks after inoculation. Negative staining of the fresh inoculated Nicotiana tabacum cv. Coker showed the presence of membrane-bound spherical particles with an average diameter of 94.25nm under transmission electron microscope. The shape and size of the particles were similar to tospovirus. SDS-PAGE analysis of partial purified virions from inoculated N. tabacum produced a strong and a faint protein bands with molecular mass of approximately 29 kDa and 55 kDa. Partial purified virions of symptomatic pineapple leaves from field showed bands with molecular mass of approximately 29 kDa, 39 kDa and 55kDa. These bands may indicate the nucleocapsid protein identity of tospovirus. Furthermore, a handheld sensor, Greenseeker, was used to detect red tip symptoms on pineapple non-destructively based on spectral reflectance, measured as Normalized Difference Vegetation Index (NDVI). Red tip severity was estimated and correlated with NDVI. Linear regression models were calibrated and tested developed in order to estimate red tip disease severity based on NDVI. Results showed a strong positive relationship between red tip disease severity and NDVI (r= 0.84).

Keywords: pineapple, diagnosis, virus, NDVI

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Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo

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Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.

Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell

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Authors: Kunal Garg, Louise Theusen Hermansan, Kanoktip Puttaraska, Oliver Hendricks, Heidi Pirttinen, Leona Gilbert

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The Germ Theory (one infectious determinant is equal to one disease) has unarguably evolved our capability to diagnose and treat infectious diseases over the years. Nevertheless, the advent of technology, climate change, and volatile human behavior has brought about drastic changes in our environment, leading us to question the relevance of the Germ Theory in our day, i.e. will vector-borne disease (VBD) sufferers produce multiple immune responses when tested for multiple microbes? Vector diseased patients producing multiple immune responses to different microbes would evidently suggest human polymicrobial infections (HPI). Ongoing diagnostic tools are exceedingly unequipped with the current research findings that would aid in diagnosing patients for polymicrobial infections. This shortcoming has caused misdiagnosis at very high rates, consequently diminishing the patient’s quality of life due to inadequate treatment. Equipped with the state-of-art scientific knowledge, PolyScan intends to address the pitfalls in current VBD diagnostics. PolyScan is a multiplex and multifunctional enzyme linked Immunosorbent assay (ELISA) platform that can test for numerous VBD microbes and allow simultaneous screening for multiple types of antibodies. To validate PolyScan, Lyme Borreliosis (LB) and spondyloarthritis (SpA) patient groups (n = 54 each) were tested for Borrelia burgdorferi, Borrelia burgdorferi Round Body (RB), Borrelia afzelii, Borrelia garinii, and Ehrlichia chaffeensis against IgM and IgG antibodies. LB serum samples were obtained from Germany and SpA serum samples were obtained from Denmark under relevant ethical approvals. The SpA group represented chronic LB stage because reactive arthritis (SpA subtype) in the form of Lyme arthritis links to LB. It was hypothesized that patients from both the groups will produce multiple immune responses that as a consequence would evidently suggest HPI. It was also hypothesized that the multiple immune response proportion in SpA patient group would be significantly larger when compared to the LB patient group across both antibodies. It was observed that 26% LB patients and 57% SpA patients produced multiple immune responses in contrast to 33% LB patients and 30% SpA patients that produced solitary immune responses when tested against IgM. Similarly, 52% LB patients and an astounding 73% SpA patients produced multiple immune responses in contrast to 30% LB patients and 8% SpA patients that produced solitary immune responses when tested against IgG. Interestingly, IgM immune dysfunction in both the patient groups was also recorded. Atypically, 6% of the unresponsive 18% LB with IgG antibody was recorded producing multiple immune responses with the IgM antibody. Similarly, 12% of the unresponsive 19% SpA with IgG antibody was recorded producing multiple immune responses with the IgM antibody. Thus, results not only supported hypothesis but also suggested that IgM may atypically prevail longer than IgG. The PolyScan concept will aid clinicians to detect patients for early, persistent, late, polymicrobial, & immune dysfunction conditions linked to different VBD. PolyScan provides a paradigm shift for the VBD diagnostic industry to follow that will drastically shorten patient’s time to receive adequate treatment.

Keywords: diagnostics, immune dysfunction, polymicrobial, TICK-TAG

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Authors: Rodriguez Diego, Del Valle Martin, Hargreaves Matias, Riveros Jose Luis

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Nutrient deficiencies due to a diet low in quantitative and qualitative terms, are prevalent throughout the developing world, especially in sub-Saharan Africa. Children and women of childbearing age are especially vulnerable. Limited availability, access and intake of animal foods at home and lack of knowledge about their value in the diet and the role they play in health, contribute to poor diet quality. Poor bioavailability of micronutrients in diets based on foods high in fiber and phytates, the low content of some micronutrients in these foods are further factors to consider. Goats are deeply embedded in almost every Sub-Saharan African rural culture, generally kept for their milk, meat, hair or leather. Goats have played an important role in African social life, especially in food security. Goat meat has good properties for human wellbeing, with a special role in lower income households. It has a high-quality protein (20 protein g/100 meat g) including all essential amino acids, good unsaturated/satured fatty acids relationship, and it is an important B-vitamin source with high micronutrients bioavailability. Mozambique has major food security problems, with poor food access and utilization, undiversified diets, chronic poverty and child malnutrition. Our objective was to design a nutritional intervention based on a dietary diversification, nutritional education, cultural beliefs and local resources, aimed to strengthen food security of children at Barrio Broma village (15°43'58.78"S; 32°46'7.27"E) in Chitima, Mozambique. Two surveys were conducted first of socio-productive local databases and then to 100 rural households about livelihoods, food diversity and anthropometric measurements in children under 5 years. Our results indicate that the main economic activity is goat production, based on a native breed with two deliveries per year in the absence of any management. Adult goats weighted 27.2±10.5 kg and raised a height of 63.5±3.8 cm. Data showed high levels of poverty, with a food diversity score of 2.3 (0-12 points), where only 30% of households consume protein and 13% iron, zinc, and B12 vitamin. The main constraints to food security were poor access to water and low income to buy food. Our dietary intervention was based on improving diet quality by increasing the access to dried goat meat, fresh vegetables, and legumes, and its utilization by a nutritional education program. This proposal was based on local culture and living conditions characterized by the absence of electricity power and drinkable water. The drying process proposed would secure the food maintenance under local conditions guaranteeing food safety for a longer period. Additionally, an ancient local drying technique was rescued and used. Moreover, this kind of dietary intervention would be the most efficient way to improve the infant nutrition by delivering macro and micronutrients on time to these vulnerable populations.

Keywords: child malnutrition, dietary diversification, food security, goat meat

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Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

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Authors: Virve Pääkkönen, Heidi M. Cuffaro, Leo Tjäderhane

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Objectives: Odontoblasts are the outermost cell layer of dental pulp and form the dentin. Importance of bacterial products, e.g. lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria and lipopolysaccharide (LPS), a cell wall component of Gram-negative bacteria, have been indicated in the pathogenesis of pulpitis. Gram-positive bacteria are more prevalent in superficial carious lesion while the amount gram-negative is higher in the deep lesions. Objective of this study was to investigate the effect of these bacterial products on inflammatory response of pulp tissue. Interleukins (IL) were of special interest. Various ILs have been observed in the dentin-pulp complex of carious tooth in vivo. Methods: Tissue culture method was used for testing the effect of LTA and LPS on human odontoblasts. Enzymatic isolation technique was used to extract living odontoblasts for cell cultures. DNA microarray and quantitative PCR (qPCR) were used to characterize the changes in the expression profile of the tissue cultured odontoblasts. Laser microdissection was used to cut healthy and affected dentin and odontoblast layer directly under carious lesion for experiments. Cytokine array detecting 80 inflammatory cytokines was used to analyze the protein content of conditioned culture media as well as dentin and odontoblasts from the carious teeth. Results: LPS caused increased gene expression IL-1α, and -8 and decrease of IL-1β, 12 , -15 and -16 after 1h treatment, while after 24h treatment decrease of IL-8, -11 and 23 mRNAs was observed. LTA treatment caused cell death in the tissue cultured odontoblasts but in in the cell culture but not in cell culture. Cytokine array revealed at least 2-fold down-regulation of IL-1β, -10 and -12 in response to LPS treatment. Cytokine array of odontoblasts of carious teeth, as well as LPS-treated tissue-cultured odontoblasts, revealed increased protein amounts of IL-16, epidermal growth factor (EGF), angiogenin and IGFBP-1 as well as decreased amount of fractalkine. In carious dentin, increased amount of IL-1β, EGF and fractalkine was observed, as well as decreased level of GRO-1 and HGF. Conclusion: LPS caused marked changes in the expression of inflammatory cytokines in odontoblasts. Similar changes were observed in the odontoblasts cut directly under the carious lesion. These results help to shed light on the inflammatory processes happening during caries.

Keywords: inflammation, interleukin, lipoteichoic acid, odontoblasts

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Authors: Alireza Vakili, Shahab Ehtesham, Mohsen Danesh Mesgaran

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The objective of this study was use different levels of spent mushroom substrate as a suitable substitute for wheat straw in the ration of male lambs. In this study 20 male lambs with the age of 90 days and initial average weight of 33± 1.7 kg were used. The animals were divided separately into single boxes with four treatments (control treatment, spent mushroom substrate 15%, spent mushroom substrate 25% and spent mushroom substrate 35%) and five replications. The experiment period was 114 days being 14 days adaptation and 90 days for breeding. On the days 36 and 94, blood samples were taken from the jugular vein. In order to carry out the trial, 20 male lambs received the four experimental diets in completely randomized design. The statistical analyses were carried out by using the GLM procedure of SAS 9.1. Means among treatments were compared by Tukey test. The results of the study showed that there was no significant differences between the serum biochemical and hematological contents of the lambs in the four treatments (p>0.05). It was concluded that spent mushroom substrate consumption has no harmful effect on the blood parameters of Kurdish male lambs.

Keywords: alternative food, nutrition, sheep performance, spent mushroom substrate

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Authors: Armaghan Moravej Aleali, Seyed Bahman Ghaderian, Homeira Rashidi, Mahmoud Mapar

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Background and Objective The metabolic syndrome (MetS) is considered the most important public health threat of the 21st century. This syndrome is characterized by a cluster of cardiovascular risk factors including increased central abdominal obesity, elevated triglycerides, reduced high-density lipoprotein, high blood pressure, increased fasting glucose, and hyperinsulinemia. MetS has been associated with hypogonadism and erectile dysfunction (ED), and MetS may be considered a risk factor for ED. The aim of this study was finding an association between metabolic syndrome and hypogonadism in Khouzestan, Iran. Subjects and Methods: In this study, 60 patients divided into two groups consisted of 30 cases (with metabolic syndrome) and 30 controls. Total and free Serum Testosterone and FBS in all of them were measured. Data was analyzed with SPSS20 program. Results: There was a significant difference between two groups about free Testosterone (P=0.01), FBS (P=0.002) and LH (P=0.03). Conclusion: According to this finding, it is thought the prevalence of hypogonadism in men with metabolic syndrome is more than the general population.

Keywords: metabolic syndrome, fasting blood sugar, hypogonadism, testosterone

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Authors: K. Makangali, G. Tokysheva, A. Shoman

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Goat meat and goat meat products have garnered increasing attention within the realm of nutrition and health due to their potential to provide a myriad of benefits. This scientific article presents a comprehensive review of the health advantages associated with goat meat consumption and the products derived from it. The paper explores the nutritional content of goat meat, highlighting its favorable composition in terms of protein, essential minerals, and amino acids. It delves into the intricate balance of macronutrients, with lower fat and cholesterol levels compared to other meats, making goat meat a desirable choice for individuals seeking healthier dietary options.

Keywords: goat meat, amino acid, nutrition, meat products, meat

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Authors: Wael Ali Mohammed Hadi

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Pseudomonas fluorescens B29B, which has asparaginase enzymatic activity, was isolated from the surface coastal seawater of Trivandrum, India. We report the complete Pseudomonas fluorescens B29B genome sequenced, identified, and annotated from a marine source. We find the genome at most minuscule a 7,331,508 bp single circular chromosome with a GC content of 62.19% and 6883 protein-coding genes. Three hundred forty subsystems were identified, including two predicted asparaginases from the genome analysis of P. fluorescens B29B for further investigation. This genome data will help further industrial biotechnology applications of proteins in general and asparaginase as a target.

Keywords: pseudomonas, marine, asparaginases, Kerala, whole-genome

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Authors: Shwu-Ling Peng, Chiung-Man Tsai, Chia-Jui Weng

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Chronic micro-inflammation is a hallmark of many aging-related neurodegenerative and metabolic syndrome-driven diseases. In high glucose (HG) environment, reactive oxygen species (ROS) is generated and the ROS induced inflammation, cytokines secretion, DNA damage, and cell cycle arrest to lead to cellular senescence. Water chestnut shell (WCS) is a plant hull which containing polyphenolic compounds and showed antioxidant and anticancer activities. Orchid, which containing a natural polysaccharide compound, possesses many physiological activities including anti-inflammatory and neuroprotective effects. These agricultural plants might be able to reduce oxidative stress and inflammation. This study was used HG-induced human normal dermal fibroblasts (HG-HNDFs) as an in vitro model to disclose the effects of water extract of Phalaenopsis orchid flower (WEPF) and ethanol extract of water chestnut shell (EEWCS) on the anti-aging and their underlying molecular mechanisms. The toxicity of extracts on human normal dermal fibroblasts (HNDFs) was determined by MTT method. The senescence of cells was assayed by β-galactosidase (SA-β-gal) kit. ROS and nitrate production was analyzed by Intracellular ROS contents and ELISA, respectively. Western blotting was used to detect the proteins in cells. The results showed that the exposure of HNDFs to HG (30 mM) for 72 h were caused cellular senescence and arrested cells at G0/G1 phase. Indeed, the treatment of HG-HNDFs with WEPF (200 μg/ml) and EEWCS (10 μg/ml) significantly released cell cycle arrest and promoted cell proliferation. The G1/S phase transition regulatory proteins such as protein retinoblastoma (pRb), p53, and p16ᴵᴺᴷ⁴ᵃ depressed by WEPF and EEWCS were also observed. Additionally, the treatment of WEPF and EEWCS increased the activity of HO-1 through upregulating Nrf2 as well as decreased the ROS and NO of HG-HNDFs. Therefore, the senescence marker protein-30 (SMP30) in cells was diminished. In conclusion, the WEPF and EEWCS might inhibit HG-induced aging of HNDFs by reducing oxidative stress and free radicals.

Keywords: agricultural plant extract, anti-aging, high glucose, Phalaenopsis orchid flower, water chestnut shell

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Authors: Mohamed M. Ibrahim, Gaber A. M. Mersal, Salih Al-Juaid, Samir A. El-Shazly

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A series of mixed ligand complexes, viz., [Cu(BPy)(Gly)X]Y {X = Cl (1), Y = 0; X = 0, Y = ClO4- (2); X = H2O, Y = NO3- (3); X = H2O, Y = CH3COO- (4); and [Cu(BPy)(Gly)-(H2O)]2(SO4) (5) have been synthesized. Their structures and properties were characterized by elemental analysis, thermal analaysis, IR, UV–vis, and ESR spectroscopy, as well as electrochemical measurements including cyclic voltammetry, electrical molar conductivity, and magnetic moment measurements. Complexes 1 and 2 formed slightly distorted square-pyramidal coordination geometries of CuN3OCl and CuN3O2, respectively in which the N,O-donor glycine and N,N-donor bipyridyl bind at the basal plane with chloride ion or water as the axial ligand. Complex 3 shows square planar CuN3O coordination geometry, which exhibits chemically significant hydrogen bonding interactions besides showing coordination polymer formation. The superoxide dismutase and catalase-like activities of all complexes were tested and were found to be promising candidates as durable electron-transfer catalyst being close to the efficiency of the mimicking enzymes displaying either catalase or tyrosinase activity to serve for complete reactive oxygen species (ROS) detoxification, both with respect to superoxide radicals and related peroxides. The DNA binding interaction with super coiled pGEM-T plasmid DNA was investigated by using spectral (absorption and emission) titration and electrochemical techniques. The results revealed that DNA intercalate with complexes 1 and 2 through the groove binding mode. The calculated intrinsic binding constant (Kb) of 1 and 2 were 4.71 and 2.429 × 105 M−1, respectively. Gel electrophoresis study reveals the fact that both complexes cleave super coiled pGEM-T plasmid DNA to nicked and linear forms in the absence of any additives. On the other hand, the interaction of both complexes with DNA, the quasi-reversible CuII/CuI redox couple slightly improves its reversibility with considerable decrease in current intensity. All the experimental results indicate that the bipyridyl mixed copper(II) complex (1) intercalate more effectively into the DNA base pairs.

Keywords: enzyme mimics, mixed ligand complexes, X-ray structures, antioxidant, DNA-binding, DNA cleavage

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Authors: Sahar B. Ahmed, M. Mostafa Said, Mona I. Mohamed

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Streptozotocin (STZ) caused a significant increase in blood glucose and malondialdehyde (MDA) levels in serum accompanied by a significant decrease in blood reduced glutathione (GSH) and superoxide dismutase (SOD) activities. Also, ALT, AST, albumin and urea were markedly affected by STZ injection. The oral administration of Mango leaves extract (MLE) one hour before STZ injection was significantly improved the blood glucose level, ALT, AST activities, albumin and urea that associated with the regulation of MDA, GSH and SOD levels. The antimicrobial activity of MLE showed a significant inhibitory activity against multidrug resistant gram positive and gram negative bacteria isolated from patients in Egyptian hospitals especially Salmonella typhi and typhimurium. In conclusion, results revealed the antioxidant, hypoglycemic effect and antimicrobial potentials of MLE under investigation. Further studies will be needed to investigate the prolonged period of MLE administration and its possible side effects.

Keywords: aqueous extract of mango leaves, STZ, antioxidant, hypoglycemic effect, antimicrobial potentials.

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Authors: Burak Dik, Oguzhan Avci, Irmak Dik, Emre Bahcivan

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The objective of this study was to evaluate the effects of alone and combined administration of Peste des petits ruminants (PPR) vaccine with Corynebacterium cutis lysate (CCL) on the hematology and biochemistry parameters levels in sheep. CCL and PPR vaccine changes cell and organ activity. In this study, 12 ewes were divided into equal groups; first group; PPR vaccine was applied only one time 1 mL subcutan of armpit on 6 sheep, and the second group; CCL (1 mL) and PPR vaccine (1 mL) combination were applied only one time subcutan of armpit on 6 sheep. Blood samples were collected before treatment (0. hour, control) and after treatment (1, 3, 7, 14, 21 and 28 days) from the sheep. Plasma and serum samples were evaluated for hematology and biochemistry parameters and there were statistically significant in sheep. In conclusion, combined usage of PPR vaccine with CCL may not influence cells and organs. Repeated CCL treatment with vaccine can create hepatotoxic, renal and bone marrow effects in sheep.

Keywords: Corynebacterium cutis lysate, hematology, peste des petits ruminants, vaccine

Procedia PDF Downloads 251

Authors: Henri El Zakhem, Dona Shalhoub, Elias Atallah, Jessica Koura

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Due to the increase of demand on dietetic food and the need for more types of diet food, the production of dietetic food is increasing and improving. This demand on dietetic food has triggered us to study the market in which we found that Kishk (Lebanese dairy product) diet is not available. Production of a low fat product which is diet Kishk was our concern. A strategy was followed to choose the right idea that will satisfy the need of the market. The whole process was studied and explained thoroughly. The percentage of fat was found to be 32.52 % in regular Kishk and 3.84 % in the diet Kishk produced. The new product has the advantage to be high in protein, low in fat.

Keywords: design and industrialization, dietetic, diet Kishk, fat

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Authors: S. Al-Khalasi, O. Mahgoub, H. Yaakub

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This study evaluated the use of raw or processed Prosopis juliflora (Meskit) pods as a major ingredient in a formulated ration to provide an alternative non-conventional concentrate for livestock feeding in Oman. Dry Meskit pods were reduced to lengths of 0.5- 1.0 cm to ensure thorough mixing into three diets. Meskit pods were subjected to two types of treatments; roasting and soaking. They were roasted at 150оC for 30 minutes using a locally-made roasting device (40 kg barrel container rotated by electric motor and heated by flame gas cooker). Chopped pods were soaked in tap water for 24 hours and dried for 2 days under the sun with frequent turning. The Meskit-pod-based diets (MPBD) were formulated and pelleted from 500 g/kg ground Meskit pods, 240 g/kg wheat bran, 200 g/kg barley grain, 50 g/kg local dried sardines and 10 g/kg of salt. Twenty four 10 months-old intact Omani male lambs with average body weight of 27.3 kg (± 0.5 kg) were used in a feeding trial for 84 days. They were divided (on body weight basis) and allocated to four diet combination groups. These were: Rhodes grass hay (RGH) plus a general ruminant concentrate (GRC); RGH plus raw Meskit pods (RMP) based concentrate; RGH plus roasted Meskit pods (ROMP) based concentrate; RGH plus soaked Meskit pods (SMP) based concentrate Daily feed intakes and bi-weekly body weights were recorded. MPBD had higher contents of crude protein (CP), acid detergent fibre (ADF) and neutral detergent fibre (NDF) than the GRC. Animals fed various types of MPBD did not show signs of ill health. There was a significant effect of feeding ROMP on the performance of Omani sheep compared to RMP and SMP. The ROMP fed animals had similar performance to those fed the GRC in terms of feed intake, body weight gain and feed conversion ratio (FCR).This study indicated that roasted Meskit pods based diet may be used instead of the commercial concentrate for feeding Omani sheep without adverse effects on performance. It offers a cheap alternative source of protein and energy for feeding Omani sheep. Also, it might help in solving the spread impact of Meskit trees, maintain the ecosystem and helping in preserving the local tree species.

Keywords: growth, Meskit, Omani sheep, Prosopis juliflora

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Authors: Caroline Provost, Francois Dumont

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Production of insects for human and animal consumption is an increasingly important activity in Canada. Protein production is more efficient and less harmful to the environment using insect rearing compared to the impact of traditional livestock, poultry and fish farms. Insects are rich in essential amino acids, essential fatty acids and trace elements. Thus, insect-based products could be used as a food supplement for livestock and domestic animals and may even find their way into the diets of high performing athletes or fine dining. Nevertheless, several parameters remain to be determined to ensure efficient and profitable production that meet the potential of these sectors. This project proposes to improve the production processes, rearing diets and processing methods for three species with valuable gastronomic and nutritional potential: the common mealworms (Tenebrio molitor), the small mealworm (Alphitobius diaperinus), and the giant mealworm (Zophobas morio). The general objective of the project is to acquire specific knowledge for mass rearing of insects dedicated to animal and human consumption in order to respond to current market opportunities and meet a growing demand for these products. Mass rearing of the three species of mealworm was produced to provide the individuals needed for the experiments. Mealworms eat flour from different cereals (e.g. wheat, barley, buckwheat). These cereals vary in their composition (protein, carbohydrates, fiber, vitamins, antioxidant, etc.), but also in their purchase cost. Seven different diets were compared to optimize the yield of the rearing. Diets were composed of cereal flour (e.g. wheat, barley) and were either mixed or left alone. Female fecundity, larvae mortality and growing curves were observed. Some flour diets have positive effects on female fecundity and larvae performance while each mealworm was found to have specific diet requirements. Trade-offs between mealworm performance and costs need to be considered. Experiments on the effect of flour composition on several parameters related to performance and nutritional and gastronomic value led to the identification of a more appropriate diet for each mealworm.

Keywords: mass rearing, mealworm, human consumption, diet

Procedia PDF Downloads 147

Authors: Debamitra Chakravorty, Pratap K. Parida

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Cold-adapted proteases enhance wash performance in low-temperature laundry resulting in a reduction in energy consumption and wear of textiles and are also used in the dehairing process in leather industries. Unfortunately, the possible drawbacks of using cold-adapted proteases are their instability at higher temperatures. Therefore, proteases with broad temperature stability are required. Unfortunately, wild-type cold-adapted proteases exhibit instability at higher temperatures and thus have low shelf lives. Therefore, attempts to engineer cold-adapted proteases by protein engineering were made previously by directed evolution and random mutagenesis. The lacuna is the time, capital, and labour involved to obtain these variants are very demanding and challenging. Therefore, rational engineering for cold stability without compromising an enzyme's optimum pH and temperature for activity is the current requirement. In this work, mutations were rationally designed with the aid of high throughput computational methodology of network analysis, evolutionary conservation scores, and molecular dynamics simulations for Savinase from Bacillus lentus with the intention of rendering the mutants cold stable without affecting their temperature and pH optimum for activity. Further, an attempt was made to incorporate a mutation in the most stable mutant rationally obtained by this method to introduce oxidative stability in the mutant. Such enzymes are desired in detergents with bleaching agents. In silico analysis by performing 300 ns molecular dynamics simulations at 5 different temperatures revealed that these three mutants were found to be better in cold stability compared to the wild type Savinase from Bacillus lentus. Conclusively, this work shows that cold adaptation without losing optimum temperature and pH stability and additionally stability from oxidative damage can be rationally designed by in silico enzyme engineering. The key findings of this work were first, the in silico data of H5 (cold stable savinase) used as a control in this work, corroborated with its reported wet lab temperature stability data. Secondly, three cold stable mutants of Savinase from Bacillus lentus were rationally identified. Lastly, a mutation which will stabilize savinase against oxidative damage was additionally identified.

Keywords: cold stability, molecular dynamics simulations, protein engineering, rational design

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Authors: Alireza Barari, Saeed Shirali, Ahmad Abdi

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Abstract: Introduction: Physical activity may be related to male reproductive function by affecting on thehypothalamic-pituitary-gonadal(HPG) axis. Our aim was to determine the effects of 6 weeks resistance exercise on reproductive hormones, HPG axis. The hypothalamic-pituitary-gonadal (HPG) axis refers tothe effects of endocrine glands in three-level including (i) the hypothalamic releasing hormone GnRH, which is synthesized in in a small heterogenous neuronal population and released in a pulsatile fashion, (ii) the anterior pituitary hormones, follicle-stimulating hormone(FSH) and luteinizing hormone (LH) and (iii) the gonadal hormones, which include both steroid such as testosterone (T), estradiol and progesterone and peptide hormones (such as inhibin). Hormonal changes that create a more anabolic environment have been suggested to contribute to the adaptation to strength exercise. Physical activity has an extensive impact on male reproductive function depending upon the intensity and duration of the exercise and the fitness level of the individual. However, strenuous exercise represents a physical stress and inflammation changed that challenges homeostasis. Materials and methods: Sixteen male volunteered were included in a 6-week control period followed by 6 weeks of resistance training (leg press, lat pull, chest press, squat, seatedrow, abdominal crunch, shoulder press, biceps curl and triceps press down) four times per week. intensity of training loading was 60%-75% of one maximum repetition. Participants performed 3 sets of 10 repetitions. Rest periods were two min between exercises and sets. Start with warm up exercises include: The muscles relax and stretch the body, which was for 10 minutes. Body composition, VO2max and the circulating level of free testosterone (fT), luteinizing hormone (LH), follicle-stimulating hormone (FSH), sex hormone binding globulin (SHBG) and inhibin B measured prior and post 6-week intervention. The hormonal levels of each serum sample were measured using commercially available ELISA kits. Analysis of anthropometrical data and hormonal level were compared using the independent samples t- test in both groups and using SPSS (version 19). P ≤ 0.05 was considered statistically significant. Results: For muscle strength, both lower- and upper-body strength were increased significantly. Aerobic fitness level improved in trained participant from 39.4 ± 5.6 to 41.9 ± 5.3 (P = 0.002). fT concentration rise progressively in the trained group and was significantly greater than those in the control group (P = 0.000). By the end of the 6-week resistance training, serum SHBG significantly increased in the trained group compared with the control group (P = 0.013). In response to resistance training, LH, FSH and inhibin B were not significantly changed. Discussion: According to our finfings, 6 weeks of resistance training induce fat loss without any changes in body weight and BMI. A decline of 25.3% in percentage of body fat with statiscally same weight was due to increase in muscle mass that happened during resistance exercise periods . Six weeks of resistance training resulted in significant improvement in BF%, VO2max and increasing strength and the level of fT and SHBG.

Keywords: resistance, hypothalamic, pituitary, gonadal axis

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Authors: Yasser Ahmed Mahmoud Ali Helal

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The use of CAD (Computer Aided Design) technology is ubiquitous in the architecture, engineering and construction (AEC) industry. This has led to its inclusion in the curriculum of architecture schools in Nigeria as an important part of the training module. This article examines the ethical issues involved in implementing CAD (Computer Aided Design) content into the architectural education curriculum. Using existing literature, this study begins with the benefits of integrating CAD into architectural education and the responsibilities of different stakeholders in the implementation process. It also examines issues related to the negative use of information technology and the perceived negative impact of CAD use on design creativity. Using a survey method, data from the architecture department of University was collected to serve as a case study on how the issues raised were being addressed. The article draws conclusions on what ensures successful ethical implementation. Millions of people around the world suffer from hepatitis C, one of the world's deadliest diseases. Interferon (IFN) is treatment options for patients with hepatitis C, but these treatments have their side effects. Our research focused on developing an oral small molecule drug that targets hepatitis C virus (HCV) proteins and has fewer side effects. Our current study aims to develop a drug based on a small molecule antiviral drug specific for the hepatitis C virus (HCV). Drug development using laboratory experiments is not only expensive, but also time-consuming to conduct these experiments. Instead, in this in silicon study, we used computational techniques to propose a specific antiviral drug for the protein domains of found in the hepatitis C virus. This study used homology modeling and abs initio modeling to generate the 3D structure of the proteins, then identifying pockets in the proteins. Acceptable lagans for pocket drugs have been developed using the de novo drug design method. Pocket geometry is taken into account when designing ligands. Among the various lagans generated, a new specific for each of the HCV protein domains has been proposed.

Keywords: drug design, anti-viral drug, in-silicon drug design, hepatitis C virus (HCV) CAD (Computer Aided Design), CAD education, education improvement, small-size contractor automatic pharmacy, PLC, control system, management system, communication

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Authors: Karina I. Hidas, Csaba Németh, Anna Visy, Judit Csonka, László Friedrich, Ildikó Cs. Nyulas-Zeke

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Egg yolk is a popular ingredient in the food industry due to its gelling, emulsifying, colouring, and coagulating properties. Because of the heat sensitivity of proteins, egg yolk can only be heat treated at low temperatures, so its shelf life, even with the addition of a preservative, is only a few weeks. Freezing can increase the shelf life of liquid egg yolk up to 1 year, but it undergoes gelling below -6 ° C, which is an irreversible phenomenon. The degree of gelation depends on the time and temperature of freezing and is influenced by the process of thawing. Therefore, in our experiment, we examined egg yolks thawed in different ways. In this study, unpasteurized, industrially broken, separated, and homogenized liquid egg yolk was used. Freshly produced samples were frozen in plastic containers at -18°C in a laboratory freezer. Frozen storage was performed for 90 days. Samples were analysed at day zero (unfrozen) and after frozen storage for 1, 7, 14, 30, 60 and 90 days. Samples were thawed in two ways (at 5°C for 24 hours and 30°C for 3 hours) before testing. Calorimetric properties were examined by differential scanning calorimetry, where heat flow curves were recorded. Denaturation enthalpy values were calculated by fitting a linear baseline, and denaturation temperature values were evaluated. Besides, dry matter content of samples was measured by the oven method with drying at 105°C to constant weight. For statistical analysis two-way ANOVA (α = 0.05) was employed, where thawing mode and freezing time were the fixed factors. Denaturation enthalpy values decreased from 1.1 to 0.47 at the end of the storage experiment, which represents a reduction of about 60%. The effect of freezing time was significant on these values, already the enthalpy of samples stored frozen for 1 day was significantly reduced. However, the mode of thawing did not significantly affect the denaturation enthalpy of the samples, and no interaction was seen between the two factors. The denaturation temperature and dry matter content did not change significantly either during the freezing period or during the defrosting mode. Results of our study show that slow freezing and frozen storage at -18°C greatly reduces the amount of protein that can be denatured in egg yolk, indicating that the proteins have been subjected to aggregation, denaturation or other protein conversions regardless of how they were thawed.

Keywords: denaturation enthalpy, differential scanning calorimetry, liquid egg yolk, slow freezing

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