Search results for: whey protein isolated

Authors: Lista Andriyati, Nurpudji A Taslim

Abstract:

The human immunodeficiency virus (HIV) patients have potential nutritional and metabolic problems. HIV is a virus that attacks cells T helper and impairs the function of immune cells. Infected individuals gradually become immunodeficient, results in increased susceptibility to a wide range of infections such as tuberculosis (TB). Malnutrition has destructive effects on the immune system and host defense mechanisms. Effective and proper nutritional therapies are important to improve medical outcomes and quality of life, which is associated with functional improvement. A case of 38-years old man admitted to hospital with loss of consciousness and was diagnosed HIV infection and relapse lung TB with severe malnutrition, fever, oral candidiasis, anemia (6.3 g/dL), severe hypoalbuminemia (1.9 g/dL), severe hypokalemia (2.2 mmol/L), immune depletion (1085 /µL) and elevated liver enzyme (ALT 1198/AST 375 U/L). Nutritional intervention by giving 2300 kcal of energy, protein 2 g/IBW/day, carbohydrate 350 g, fat 104 g through enteral and parenteral nutrition. Supplementations administered are zinc, vitamin A, vitamin B1, vitamin B6, vitamin B12, vitamin C, vitamin D, and snakehead fish extract high content of protein albumin (Pujimin®). After 46 days, there are clinical and metabolic improvement in Hb (6.3 to 11.2 g/dL), potassium (2.2 to 3.4 mmol/L), albumin (1.9 to 2.3 g/dL), ALT 1198 to 47/AST 375 to 68 U/L) and improved awareness. In conclusion, nutritional therapy in HIV infection with adequate macronutrients and micronutrients fulfillment and immunonutrition is very important to avoid cachexia and to improve nutritional status and immune disfunction.

Keywords: HIV, hypoalbuminemia, malnutrition, tuberculosis

Procedia PDF Downloads 109

Authors: Ole-Christian Galbo Engstrøm, Erik Schou Dreier, Birthe Møller Jespersen, Kim Steenstrup Pedersen

Abstract:

Grain quality analysis is a multi-parameterized problem that includes a variety of qualitative and quantitative parameters such as grain type classification, damage type classification, and nutrient regression. Currently, these parameters require human inspection, a multitude of instruments employing a variety of sensor technologies, and predictive model types or destructive and slow chemical analysis. This paper investigates the feasibility of applying near-infrared hyperspectral imaging (NIR-HSI) to grain quality analysis. For this study two datasets of NIR hyperspectral images in the wavelength range of 900 nm - 1700 nm have been used. Both datasets contain images of sparsely and densely packed grain kernels. The first dataset contains ~87,000 image crops of bulk wheat samples from 63 harvests where protein value has been determined by the FOSS Infratec NOVA which is the golden industry standard for protein content estimation in bulk samples of cereal grain. The second dataset consists of ~28,000 image crops of bulk grain kernels from seven different wheat varieties and a single rye variety. In the first dataset, protein regression analysis is the problem to solve while variety classification analysis is the problem to solve in the second dataset. Deep convolutional neural networks (CNNs) have the potential to utilize spatio-spectral correlations within a hyperspectral image to simultaneously estimate the qualitative and quantitative parameters. CNNs can autonomously derive meaningful representations of the input data reducing the need for advanced preprocessing techniques required for classical chemometric model types such as artificial neural networks (ANNs) and partial least-squares regression (PLS-R). A comparison between different CNN architectures utilizing 2D and 3D convolution is conducted. These results are compared to the performance of ANNs and PLS-R. Additionally, a variety of preprocessing techniques from image analysis and chemometrics are tested. These include centering, scaling, standard normal variate (SNV), Savitzky-Golay (SG) filtering, and detrending. The results indicate that the combination of NIR-HSI and CNNs has the potential to be the foundation for an automatic system unifying qualitative and quantitative grain quality analysis within a single sensor technology and predictive model type.

Keywords: deep learning, grain analysis, hyperspectral imaging, preprocessing techniques

Procedia PDF Downloads 82

Authors: Kang-Hyun Leem, Myung-Gyou Kim, Hye Kyung Kim

Abstract:

Eggs have long been an important contributor to the nutritional quality of the human, and recognized as a very valuable source of proteins for human nutrition. Egg yolk is composed of various important chemical substances for human health. Growth means not only the increase of body weight but also the elongation of height and the enlargement of each organ's anatomical and morphological size. A calcium shortage causes the growth retardation on the body growth. In this study, we examined the therapeutic effects of egg yolk peptide (EYP) on the retardation of the longitudinal bone growth induced by low-calcium diet (0.05%) in adolescent rats. Low calcium diets were administrated for 15 days. During the last five days, calcium and/or vitamin D and/or EYP were administrated. The body weights, longitudinal bone growth rates, the heights of growth plates, and bone morphogenetic protein (BMP)-2 and insulin-like growth factor (IGF)-1 expressions were measured using histochemical analysis. Low calcium diets caused the significant reduction in body weight gains and the longitudinal bone growth. The heights of growth plates and the expressions of BMP-2 and IGF-1 showed the impairment of body growth as well. Calcium and/or vitamin D administration could not significantly increase the longitudinal bone growth. However, calcium, vitamin D, and EYP administration significantly increased the bone growth, the growth plate height, and BMP-2 and IGF-1 expressions. These results suggest that EYP enhances the longitudinal bone growth in the calcium and/or vitamin D deficiency and it could be a promising agent for the treatment of children suffering from malnutrition.

Keywords: egg yolk peptide, low-calcium diet, longitudinal bone growth, morphogenetic protein-2, insulin-like growth factor-1, vitamin D

Procedia PDF Downloads 428

Authors: Bin Sheng, Changhong Lu

Abstract:

Let G be a simple undirected graph with no isolated vertex. A paired dominating set of G is a dominating set which induces a subgraph that has a perfect matching. The paired domination number of G, denoted by γₚᵣ(G), is the size of its smallest paired dominating set. Goddard and Henning conjectured that γₚᵣ(G) ≤ 4n/7 holds for every graph G with δ(G) ≥ 3, except the Petersen Graph. In this paper, we prove this conjecture for cubic graphs.

Keywords: paired dominating set, upper bound, cubic graphs, weight function

Procedia PDF Downloads 226

Authors: S. Baississe, S. Godbane, A. Lekbir

Abstract:

The objective of our work is to develop an ice cream from a fermented cream, skim milk and other ingredients and follow the evolution of its physicochemical properties, biochemical and microstructure of the products obtained. Our cream is aerated with the manufacturing steps start with a homogenizing follow different ingredients by heating to 40°C emulsion, the preparation is then subjected to a heat treatment at 65°C for 30 min, before being stored in the cold at 4°C for a few hours. This conservation promotes crystallization of the material during the globular stage of maturation of the cream. The emulsifying agent moves gradually absorbed on the surface of fat globules homogeneous, which results in reduced protein stability. During the expansion, the collusion of destabilizing fat globules in the aqueous phase favours their coalescence. During the expansion, the collusion of destabilized fat globules in the aqueous phase favours their coalescence. The stabilizing agent increases the viscosity of the aqueous phase and the drainage limit interaction with the proteins of the aqueous phase and the protein absorbed on fat globules. The cutting improved organoleptic property of our cream is made by the use of three dyes and aromas. The products obtained undergo physicochemical analyses (pH, conductivity and acidity), biochemical (moisture, % dry matter and fat in %), and finally in the microscopic observation of the microstructure and the results obtained by analysis of the image processing software. The results show a remarkable evolution of physicochemical properties (pH, conductivity and acidity), biochemical (moisture, fat and non-fat) and microstructure of the products developed in relation to the raw material (skim milk) and the intermediate product (fermented cream).

Keywords: ice cream, sour cream, physicochemical, biochemical, microstructure

Procedia PDF Downloads 188

Authors: O. B. Oriolowo, O. J. John

Abstract:

Empirical findings have shown insects to be nutritious and good source of food for man. However, human food preferences are not only determined by nutritional values of food consumed but, more importantly, by sociology and economic pressure. This study examined the interrelation between science and sociology in sustaining the acceptance of entomophagy among college students to combat food insecurity. A twenty items five Likert scale, College Students Entomophagy Questionnaire (CSEQ), was used to elucidate information from the respondents. The reliability coefficient was obtained to be 0.91 using Spearman-Brown Prophecy formula. Three research questions and three hypotheses were raised. Also, quantitative nutritional analysis of few insects and some established conventional protein sources were undertaking in order to compare their nutritional status. The data collected were analyzed using descriptive statistics of percentages and inferential statistics of correlation and Analysis of Variance (ANOVA). The results obtained showed that entomophagy has cultural heritage among different tribes in Nigeria and is an acceptable practice; it cuts across every social stratum and is practiced among both major religions. Moreover, insects compared favourably in term of nutrient contents when compared with the conventional animal protein sources analyzed. However, there is a gradual decline in the practice of entomophagy among students, which may be attributed to the influence of western civilization. This study, therefore, recommended an intensification of research and enlightenment of people on the usefulness of entomophagy so as to preserve its cultural heritage as well as boost human food security.

Keywords: entomophagy, food security, malnutrition, poverty alleviation, sociology

Procedia PDF Downloads 103

Authors: A. Abbas Khalaf, L. Issazadeh, Z. Arif Abdullah, J. Hassanpour

Abstract:

In order to evaluate the growth and yield properties of two Sorghum-Sudangrass hybrids under different irrigation levels, an investigation was done in the experiment site of Collage of Agriculture, University of Duhok, Kurdistan region of Iraq (36°5´38^⸗ N, 42°52´02^⸗ E) in the years 2015-16. The experiment was conducted under Randomized Complete Block Design (RCBD) with three replications, which main factor was irrigation treatments (I₁₀₀, I₇₅ and I₅₀) according to evaporation pan class A and type of Sorghum-Sudangrass hybrids (KH12SU9001, G₁) and (KH12SU9002, G₂) were factors of subplots. The parameters studied were: plant height (cm), number of green leaves per plant; leaf area (m²/m²), stem thickness (mm), percent of protein, fresh and dry biomass (ton.ha^-1) and also crop water productivity. The results of variance analysis showed that KH12SU9001 variety had more amount of leaf area, percent of protein, fresh and dry biomass yield in comparison to KH12SU9002 variety. By comparing effects of irrigation levels on vegetative growth and yield properties, results showed that amount of plant height, fresh and dry biomass weight was decreased by decreasing irrigation level from full irrigation regime to 5 _o% of irrigation level. Also, results of crop water productivity (CWP) indicated that improvement in quantity of irrigation would impact fresh and dry biomass yield significantly. Full irrigation regime was recorded the highest level of CWP (1.28-1.29 kg.m^-3).

Keywords: deficit irrigation, growth, sorghum-sudangrass hybrid, yield

Procedia PDF Downloads 120

Authors: T. I. Mbata, M. J. Ikenebomeh

Abstract:

Background: The study was aim at formulating a complementary foods based on maize and bambara groundnut with a view of reducing malnutrition in low income families. Protein-energy malnutrition is a major health challenge attributed to the inappropriate complementary feeding practices, low nutritional quality of traditional complementary foods and high cost of quality protein-based complementary foods. Methods: The blends 70% maize, 30% bambara groundnut were evaluated for proximate analyses, minerals, amino acids profile, and antinutritional factors, using proprietary formula (‘Nutrend’) as standard. Antinutritional factors, amino acids, microbiological properties and sensory attributes were determined using standard methods. Results; For Protein, the results were 15.0% for roasted bambara groundnut maize germinated flour (RBMGF), 13.80% for cooked bambara groundnut maize germinated flour (CBMGF), 15.18% for soaked bambara groundnut maize germinated flour (SBMGF); values for maize flour and nutrend had 10.4% and 23.21% respectively. With respect to energy value, RBMGF, CBMGF, SBMGF, maize flour and nutrend had 494.9, 327.58, 356.49, 366.8 and 467.2 kcal respectively. The percentages of total essential amino acids in the composition of the blends were 36.9%, 40.7% and 38.9% for CBMGF, SBMGF and RBMGF, respectively, non-essential amino acids contents were 63.1%, 59.3% and 61.1% for CBMGF, SBMGF and RBMGF respectively. The mineral content, that is, calcium, potassium, magnesium and sodium, of formulated samples were higher than those obtained for maize flour and Nutrend. The antinutrient composition of RBMGF and CBMGF were lower than of SBMGF. The rats fed with the control diet exhibited better growth performance such as feed intake (1527 g) and body weight gain (93.8 g). For the microbial status, microflora gradually changed from gram negative enteric bacteria, molds, lactic acid bacteria and yeast to be dominated by gram positive lactic acid bacteria (LAB) and yeasts. Yeasts and LAB growth counts in the complementary food varied between 4.44 and 7.36 log cfu/ml. LAB number increased from 5.40 to 7.36 log cfu/ml during fermentation. Yeasts increased from 4.44 to 5.60 log cfu/ml. Organoleptic evaluation revealed that the foods were well accepted. Conclusion: Based on the findings the application of bambara groundnut fortification to traditional foods can promote the nutritional quality of African maize - based traditional foods with acceptable rheological and cooking qualities.

Keywords: bambara groundnut, maize, fortification, complementary food

Procedia PDF Downloads 342

Authors: Agbugui M. O., Inobeme A.

Abstract:

Fish constitutes a vital component of human diets due to their rich nutritional compositions. They serve as a remarkable source of proteins, vitamins, and fatty acids, which are indispensable for the effective growth and development of humans. The need to explore the nutritional compositions of various species of fish in different water bodies becomes paramount. Presently, consumer concern is not just on food's nutritional value but also on the safety level. Environmental contamination by heavy metals has become an issue of pressing concern in recent times. Heavy metals, due to their ubiquitous nature, are found in various water bodies as they are released from various anthropogenic activities. This work investigated the proximate compositions, mineral contents, and heavy metals concentrations of four different species of fish (P. annectens, L. niloticus, G. niloticus, and H. niloticus) collected from the lower Niger at Agenebode using standard procedures. The highest protein contents were in Gymnarchus niloticus (37.32%), while the least was in Heterotis niloticus (20.41%). Protopterus annectens had the highest carbohydrate content (34.55%), while Heterotis niloticus had the least (12.24%). The highest lipid content (14.41%) was in Gymnarchus niloticus. The highest concentration of potassium was 21.00 ppm. The concentrations of heavy metals in ppm ranged from 0.01 – 1.4 (Cd), 0.07 – 2.89 (Pb), 0.02 – 16.4 (Hg), 0.88 – 5.1 (Cu) and 1.2 – 8.23 (Zn). The concentrations of Hg, Cd and Pb in some of the samples investigated were higher than the permissible limits based on international standards. There is a pressing need for further study focusing on various species of animals and plants in the area due to the alarming contents of these metals; remedial measures could also be ensured for safety.

Keywords: trace metals, nutritional value, human health, crude protein, lipid content

Procedia PDF Downloads 77

Authors: Osama K. Abou-Zied, Saba A. Sulaiman

Abstract:

We studied the spectroscopy of 25-nm diameter gold nanoparticles (AuNPs), coated with human serum albumin (HSA) as a model drug carrier. The morphology and coating of the AuNPs were examined using transmission electron microscopy and dynamic light scattering. Resonance energy transfer from the sole tryptophan of HSA (Trp214) to the AuNPs was observed in which the fluorescence quenching of Trp214 is dominated by a static mechanism. Using fluorescein (FL) to probe the warfarin drug-binding site in HSA revealed the unchanged nature of the binding cavity on the surface of the AuNPs, indicating the stability of the protein structure on the metal surface. The transient absorption results of the surface plasmonic resonance (SPR) band of the AuNPs show three ultrafast dynamics that are involved in the relaxation process after excitation at 460 nm. The three decay components were assigned to the electron-electron (~ 400 fs), electron-phonon (~ 2.0 ps) and phonon-phonon (200–250 ps) interactions. These dynamics were not changed upon coating the AuNPs with HSA which indicates the chemical and physical stability of the AuNPs upon bioconjugation. Binding of FL in HSA did not have any measurable effect on the bleach recovery dynamics of the SPR band, although both FL and AuNPs were excited at 460 nm. The current study is important for a better understanding of the physical and dynamical properties of protein-coated metal nanoparticles which are expected to help in optimizing their properties for critical applications in nanomedicine.

Keywords: gold nanoparticles, human serum albumin, fluorescein, femtosecond transient absorption

Procedia PDF Downloads 313

Authors: Katarína Čurová, Leonard Siegfried, Radka Vargová, Marta Kmeťová, Vladimír Hrabovský

Abstract:

Introduction: Cytolethal distending toxins (CDTs) represent intracellular acting proteins which interfere with cell cycle of eukaryotic cells. They are produced by Gram-negative bacteria with afinity to mucocutaneous surfaces and could play a role in the pathogenesis of various diseases. CDTs induce DNA damage probably through DNAse activity, which causes cell cycle arrest and leads to further changes (cell distension and death, apoptosis) depending on the cell type. Five subtypes of CDT (I to V) were reported in E. coli. Methods: We examined 252 E. coli strains belonging to four different groups. Of these strains, 57 were isolated from patients with diarrhea, 65 from patients with urinary tract infections (UTI), 65 from patients with sepsis and 65 from patients with other extraintestinal infections (mostly surgical wounds, decubitus ulcers and respiratory tract infections). Identification of these strains was performed by MALDI-TOF analysis and detection of genes encoding CDTs and determination of the phylogenetic group was performed by PCR. Results: In this study, we detected presence of cdt genes in 11 of 252 E. coli strains tested (4,4 %). Four cdt positive E. coli strains were confirmed in group of UTI (6,15 %), three cdt positive E. coli strains in groups of diarrhea (5,3 %) and other extraintestinal infections (4,6 %). The lowest incidence, one cdt positive E. coli strain, was observed in group of sepsis (1,5 %). All cdt positive E. coli strains belonged to phylogenetic group B2. Conclusion: CDT-producing E. coli are isolated in a low percentage from patients with intestinal and extraintestinal infections, including sepsis and our results correspond with these studies. A weak prevalence of cdt genes suggests that CDTs are not major virulence factors but in combination with other virulence factors may increase virulence potential of E. coli. We suppose that all 11 cdt positive E. coli strains represent real pathogens because they belong to the phylogenetic group B2 which is pathogenic lineage for bacteria E. coli.

Keywords: cytolethal distending toxin, E. coli, phylogenetic group, extraintestinal infection, diarrhea

Procedia PDF Downloads 336

Authors: Esra Sengul, R. G. Aktas, M. E. Sitar, H. Isan

Abstract:

Hepatocellular carcinoma (HCC) is a hepatocellular tumor commonly found on the surface of the chronic liver. HepG2 is the most commonly used cell type in HCC studies. The main proteins remaining in the blood serum after separation of plasma fibrinogen are albumin and globulin. The fact that the albumin showed hepatocellular damage and reflect the synthesis capacity of the liver was the main reason for our use. Alpha-Fetoprotein (AFP) is an albumin-like structural embryonic globulin found in the embryonic cortex, cord blood, and fetal liver. It has been used as a marker in the follow-up of tumor growth in various malign tumors and in the efficacy of surgical-medical treatments, so it is a good protein to look at with albumins. We have seen the morphological changes of dimethyl sulfoxide (DMSO) on HepG2 and decided to investigate its biochemical effects. We examined the effects of DMSO, which is used in cell cultures, on albumin, AFP and total protein at low doses. Material Method: Cell Culture: Medium was prepared in cell culture using Dulbecco's Modified Eagle Media (DMEM), Fetal Bovine Serum Dulbecco's (FBS), Phosphate Buffered Saline and trypsin maintained at -20 ° C. Fixation of Cells: HepG2 cells, which have been appropriately developed at the end of the first week, were fixed with acetone. We stored our cells in PBS at + 4 ° C until the fixation was completed. Area Calculation: The areas of the cells are calculated in the ImageJ (IJ). Microscope examination: The examination was performed with a Zeiss Inverted Microscope. Daytime photographs were taken at 40x, 100x 200x and 400x. Biochemical Tests: Protein (Total): Serum sample was analyzed by a spectrophotometric method in autoanalyzer. Albumin: Serum sample was analyzed by a spectrophotometric method in autoanalyzer. Alpha-fetoprotein: Serum sample was analyzed by ECLIA method. Results: When liver cancer cells were cultured in medium with 1% DMSO for 4 weeks, a significant difference was observed when compared with the control group. As a result, we have seen that DMSO can be used as an important agent in the treatment of liver cancer. Cell areas were reduced in the DMSO group compared to the control group and the confluency ratio increased. The ability to form spheroids was also significantly higher in the DMSO group. Alpha-fetoprotein was lower than the values of an ordinary liver cancer patient and the total protein amount increased to the reference range of the normal individual. Because the albumin sample was below the specimen value, the numerical results could not be obtained on biochemical examinations. We interpret all these results as making DMSO a caretaking aid. Since each one was not enough alone we used 3 parameters and the results were positive when we refer to the values of a normal healthy individual in parallel. We hope to extend the study further by adding new parameters and genetic analyzes, by increasing the number of samples, and by using DMSO as an adjunct agent in the treatment of liver cancer.

Keywords: hepatocellular carcinoma, HepG2, dimethyl sulfoxide, cell culture, ELISA

Procedia PDF Downloads 122

Authors: Hafiza Javaria Ashraf, Xinghong Wang, Zhanghong Shi, Youming Hou

Abstract:

Insect’s innate immunity depends on a variety of defense responses for the recognition of invading pathogens. Pathogen recognition involves particular proteins known as pattern recognition receptors (PRRs). These PRRs interact with pathogen-associated molecular patterns (PAMPs) present on the surface of pathogens to distinguish between self and non-self. C-type lectins (CTLs) belong to a superfamily of PPRs which involved in insect immunity and defense mechanism. Rhynchophorus ferrugineus Olivier is a devastating pest of Palm cultivations in China. Although studies on R. ferrugineus immune mechanism and host defense have conducted, however, the role of CTL in immune responses of R. ferrugineus remains elusive. Here, we report RfCTL, which is a secreted protein containing a single-CRD domain. The open reading frame (ORF) of CTL is 226 bp, which encodes a putative protein of 168 amino acids. Transcript expression analysis revealed that RfCTL highly expressed in immune-related tissues, i.e., hemolymph and fat body. The abundance of RfCTL in the gut and fat body dramatically increased upon Staphylococcus aureus and Escherichia coli bacterial challenges, suggesting a role in defense against gram-positive and gram-negative bacterial infection. Taken together, we inferred that RfCTL might be involved in the immune defense of R. ferrugineus and established a solid foundation for future studies on R. ferrugineus CTL domain proteins for better understanding of insect immunity.

Keywords: biological invasion, c-type lectin, insect immunity, Rhynchophorus ferrugineus Oliver

Procedia PDF Downloads 134

Authors: Jaruwan Duangchuen, Siwalak Pathaveerat

Abstract:

Coconut (Cocos nucifera) belongs to the family Arecaceae. Coconut juice and meat are consumed as food and dessert in several regions of the world. Coconut juice contains low proteins, and arginine is the main amino acid content. Coconut meat is the endosperm of coconut that has nutritional value. It composes of carbohydrate, protein and fat. The objective of this study is utilization of by-products from the virgin coconut oil extraction process by using the skimmed coconut milk as a powder. The skimmed coconut milk was separated from the coconut milk in virgin coconut oil extraction process that consists approximately of protein 6.4%, carbohydrate 7.2%, dietary fiber 0.27 %, sugar 6.27%, fat 3.6 % and moisture content of 86.93%. This skimmed coconut milk can be made to powder for value - added product by using spray drying. The factors effect to the yield and properties of dry skimmed coconut milk in spraying process are inlet, outlet air temperature and the maltodextrin concentration. The percentage of maltodextrin content (15, 20%), outlet air temperature (80 ºC, 85 ºC, 90 ºC) and inlet air temperature (190 ºC, 200 ºC, 210 ºC) were conducted to the skimmed coconut milk spray drying process. The spray dryer was kept air flow rate (0.2698 m3 /s). The result that shown 2.22 -3.23% of moisture content, solubility, bulk density (0.4-0.67g/mL), solubility, wettability (4.04 -19.25 min) for solubility in the water, color, particle size were analyzed for the powder samples. The maximum yield (18.00%) of spray dried coconut milk powder was obtained at 210 °C of temperature, 80°C of outlet temperature and 20% maltodextrin for 27.27 second for drying time. For the amino analysis shown that the high amino acids are Glutamine (16.28%), Arginine (10.32%) and Glycerin (9.59%) by using HPLP method (UV detector).

Keywords: skimmed coconut milk, spray drying, virgin coconut oil process (VCO), maltodextrin

Procedia PDF Downloads 310

Authors: Ghibeche Abderrahmane

Abstract:

To examine whether genes associated with cellular defense against oxidative stress are associated with insulin sensitivity, patients with type 2 diabetes (n=7) and age-matched (n=5) and young (n=9) control subjects underwent a euglycemic-hyperinsulinemic clamp for 120 min. Muscle samples were obtained before and after the clamp and analyzed for heat shock protein (HSP)72 and heme oxygenase (HO)-1 mRNA, intramuscular triglyceride content, and the maximal activities of β-hyroxyacyl-CoA dehydrogenase (β-HAD) and citrate synthase (CS). Basal expression of both HSP72 and HO-1 mRNA were lower (P < 0.05) by 33 and 55%, respectively, when comparing diabetic patients with age-matched and young control subjects, with no differences between the latter groups. Both basal HSP72 (r = 0.75, P < 0.001) and HO-1 (r = 0.50,P < 0.05) mRNA expression correlated with the glucose infusion rate during the clamp. Significant correlations were also observed between HSP72 mRNA and both β-HAD (r = 0.61, P < 0.01) and CS (r = 0.65, P < 0.01). HSP72 mRNA was induced (P < 0.05) by the clamp in all groups. Although HO-1 mRNA was unaffected by the clamp in both the young and age-matched control subjects, it was increased (P < 0.05) ∼70-fold in the diabetic patients after the clamp. These data demonstrate that genes involved in providing cellular protection against oxidative stress are defective in patients with type 2 diabetes and correlate with insulin-stimulated glucose disposal and markers of muscle oxidative capacity. The data provide new evidence that the pathogenesis of type 2 diabetes involves perturbations to the antioxidant defense mechanism within skeletal muscle.

Keywords: euglycemic-hyperinsulinemic, HSP72, mRNA, diabete

Procedia PDF Downloads 423

Authors: Irma Gabriela Lopez-Moreno, Elva Perez-Luque, Herlinda Aguilar-Zavala

Abstract:

Introduction: Type 2 Diabetes Mellitus (T2DM) is characterized by combination of insulin resistance and deterioration of insulin secretion. Sfrp5 is a protein that antagonizes Wnt5a proteins by preventing it from reaching its receptor and activating the Wnt/β-catenin signaling pathway, this pathway is one of the most important regulators of adipogenesis. Although metformin decreases glucose levels its mechanisms of action are not fully known but it has been implicated in the inhibition of the Wnt/β-catenin signaling pathway. Objective: The objective was evaluating the effects of metformin on serum levels of Sfrp5 in patients with T2DM treated and not treated with metformin. Methods: Two groups of patients were selected: one group of T2DM patients treated with metformin (n = 35) and another group of subjects with recent diagnosis of T2DM untreated (n = 35) with a mean age of 48 ± 9 years. In these subjects anthropometric measures were taken as weight, height, waist and hip circumference, were calculated the percentage of body fat, visceral fat and muscle mass. In addition, were measured glucose levels, lipid profile, adiponectin and Sfrp5. Results: Sfrp5 were higher in metformin-treated patients compared to the untreated group (19.9 vs 13.6 ng/mL p < 0.001), a negative correlation was found between Sfrp5 levels and total cholesterol levels (r= -0.25, p = 0.03) and percentage of visceral fat (r = -0.26, p = 0.03) and a positive correlation with HDL cholesterol levels (r = 0.31, p = 0.01) and adiponectin (r=0.65, p = < 0.001). Conclusions: The findings show that metformin consumption increased levels of Sfrp5, which may lead to a decrease in the activation of the WNT/β-catenin pathway impacting on adipogenesis.

Keywords: adiponectin, diabetes, metformin, Sfrp5

Procedia PDF Downloads 164

Authors: Mustafa Al-Yaseen, Haider Mohammed Mahdi, Haider Ali Al–Zahid, Nazar S. Haddad

Abstract:

Background: Early diagnosis of musculoskeletal infections is of vital importance to avoid devastating complications. There is no single laboratory marker which is sensitive and specific in diagnosing these infections accurately. White blood cell count, erythrocyte sedimentation rate, and C-reactive protein are not specific as they can also be elevated in conditions other than bacterial infections. Materials Culture and sensitivity is not a true gold standard due to its varied positivity rates. Serum Procalcitonin is one of the new laboratory markers for pyogenic infections. The objective of this study is to assess the value of PCT in the diagnosis of soft tissue, bone, and joint infections. Patients and Methods: Patients of all age groups (seventy-four patients) with a diagnosis of musculoskeletal infection are prospectively included in this study. All patients were subjected to White blood cell count, erythrocyte sedimentation rate, C-reactive protein, and serum Procalcitonin measurements. A healthy non infected outpatient group (twenty-two patients) taken as a control group and underwent the same evaluation steps as the study group. Results: The study group showed mean Procalcitonin levels of 1.3 ng/ml. Procalcitonin, at 0.5 ng/ml, was (42.6%) sensitive and (95.5%) specific in diagnosing of musculoskeletal infections with (positive predictive value of 87.5% and negative predictive value of 48.3%) and (positive likelihood ratio of 9.3 and negative likelihood ratio of 0.6). Conclusion: Serum Procalcitonin, at a cut – off of 0.5 ng/ml, is a specific but not sensitive marker in the diagnosis of musculoskeletal infections, and it can be used effectively to rule in the diagnosis of infection but not to rule out it.

Keywords: procalcitonin, infection, labratory markers, musculoskeletal

Procedia PDF Downloads 149

Authors: M. Ntswane, N. Mbuma, M. Labuschagne, A. Mofokeng, M. Rantso

Abstract:

Cowpea is an essential legume for the nutrition and health of millions of people in different regions. The production and productivity of the crop are very limited in South Africa due to a lack of adapted and stable genotypes. The improvement of nutritional quality is made possible by manipulating the genes of diverse cowpea genotypes available around the world. Assessing the adaptability and stability of the cowpea mutant genotypes for yield and nutritional quality requires examining them in different environments. The objective of the study was to determine the adaptability and stability of cowpea mutant genotypes under South African conditions and to identify the superior genotypes that combine grain yield components, antioxidants, and nutritional quality. Thirty-one cowpea genotypes were obtained from the Agricultural Research Council grain crops (ARC-GC) and were planted in Glen, Mafikeng, Polokwane, Potchefstroom, Taung, and Vaalharts during the 2021/22 summer cropping season. Significant genotype by location interactions indicated the possibility of genetic improvement of these traits. The genotype plus genotype by environment indicated broad adaptability and stability of mutant genotypes. The principal component analysis identified the association of the genotypes with the traits. Phenotypic correlation analysis showed that Zn and protein content were significant and positively correlated and suggested the possibility of indirect selection of these traits. Results from this study could be used to help plant breeders in making informed decisions and developing nutritionally improved cowpea genotypes with the aim of addressing the challenges of poor nutritional quality.

Keywords: cowpea seeds, adaptability, stability, mineral elements, protein content

Procedia PDF Downloads 87

Authors: Tomáš Nejedlý, Dominika Mrázková, Jitka Viktorová

Abstract:

The blood-brain barrier (BBB) serves as a protective shield, preventing the entry of harmful substances into the central nervous system. On the other hand, it also restricts the transport of neuroactive drugs, such as antiepileptics, which mitigate epileptic seizures. Drug-resistant epilepsy is often associated with the overexpression of efflux transporters, including P-glycoprotein (P-gp) or multidrug resistance protein 1 (MRP1), on the BBB. The aim of this work is to find P-gp and MRP1 inhibitors derived from phytocannabinoids and terpenes. The work evaluates whether these compounds interact directly with P-gp or MRP1 by rhodamine 123 or fluorescein efflux assay. The effect of phytocannabinoids on the gene expression of these transporters is also studied using qPCR and Western blot. These transporters are found in BBB cells; however, we decided to use the human ovarian cancer cell line (A2780ADR) due to its overproduction of P-gp and malignant glioma cell line (U87) due to its overproduction of MRP1. The results showed that while terpenes suppressed the activity of efflux transporters, phytocannabinoids tended to decrease their expression. Terpenes demonstrated an average inhibition of 65%, surpassing phytocannabinoids, which exhibited an average inhibition of approximately 30%. Particularly noteworthy was the modulating effect of (-)-α-bisabolol with the highest activity among the compounds tested. Based on these findings, phytocannabinoids and terpenes emerge as promising natural candidates for addressing drug resistance linked to efflux transporters. Acknowledgment: The project was funded by the Grant No 22-20860S of The Czech Science Foundation.

Keywords: drug-resistant epilepsy, efflux transporters, multidrug resistance protein 1, P-glycoprotein, phytocannabinoids, terpens

Procedia PDF Downloads 44

Authors: Temitope D. Awobusuyi, Eric O. Amonsou, Muthulisi Siwela

Abstract:

Amahewu, a lactic acid fermented non-alcoholic maize based beverage is widely consumed in Southern Africa. It is traditionally made with white maize which is deficient in vitamin A. Provitamin A-biofortified maize has been developed for use as a complementary strategy to alleviate vitamin A deficiency. In this study the nutritional composition and protein digestibility of amahewu produced using provitamin A-biofortified maize was determined. Provitamin A-biofortified amahewu was processed by fermenting cooked maize porridge using malted provitamin A-biofortified maize, wheat bran and lactobacillus mixed starter culture with either malted maize or wheat bran. The total provitamin A content in amahewu products ranged from 3.3-3.8 μg/g (DW). The % retention of total provitamin A ranged from 79 %- 90 % μg/g (DW). The lowest % retention was observed in products fermented with the addition of starter culture. The gross energy of amahewu products were approx. 20 MJ/kg. There was a slight increase in the lysine content of amahewu after fermentation. Protein digestibility of amahewu (approx.91%) was slightly higher compared to unprocessed provitamin A maize (86%). However, a general decrease was observed in the minerals when compared to the unprocessed provitamin A maize. Amahewu processed using starter cultures has higher iron content than those processed with the addition of malt. These result suggests that provitamin A-biofortified amahewu has the potential to make a significant contribution towards alleviating Vitamin A Deficiency in rural communities who are also the most vulnerable to VAD.

Keywords: vitamin A deficiency, provitamin A maize, biofortification, fermentation

Procedia PDF Downloads 397

Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

Abstract:

Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

Procedia PDF Downloads 237

Authors: O. Soji-Omoniwa, N. O. Muhammad, L. A. Usman, B. P. Omoniwa

Abstract:

This study was conducted to investigate the effect of the leaf essential oil of C. sinensis harvested at 7.00a.m and 4.00p.m on some Liver and Kidney function indices of diabetic rats as well as investigate the effect of time of harvest on the observed effect. Experimental animals were divided into 4 groups (A, B, C and D). Diabetes mellitus was induced in all animals, except the normal control group (Group A), by injecting 150mg/kg body weight of alloxan monohydrate intraperitoneally. Group A received distilled water while group B (diabetic control group) was not treated. Group C and D were treated with leaf essential oil of C. sinensis harvested at 7.00 a.m and 4.00 p.m respectively at a dose of 110 mg/kg body weight every other day for 15 days. Alkaline phosphatase (ALP), Alanine Transaminase (ALT) and Aspartate Transaminase (AST) activity was evaluated in the serum, Liver and Kidney of studied animals. Total and Direct Bilirubin level, Total Protein and Globulin, Creatinine and Urea level were also evaluated. Result showed that creatinine and urea, serum ALP, AST and ALT levels was significantly reduced (p < 0.05), while the levels of total Protein and Globulin increased significantly (p < 0.05) for the treated animals compared to the diabetic control group. In conclusion, the leaf essential oil of Citrus sinensis ameliorated the impaired renal and liver function; however, the time of harvest of the leaf does not significantly affect its ameliorative effect.

Keywords: C. sinensis, function indices, harvest time, leaf essential oil.

Procedia PDF Downloads 339

Authors: Mahsa Jalili, Trude Johansen, Signe Dille Lovmo, Turid Rustad, Rolf Erik Olsen, Atle M. Bones

Abstract:

The quality of fish muscle is a key factor in fish industry, and dietary ingredients can influence fish quality. The aim of this study was to examine the impact of krill meal, soybean meal, Bactocell® and butyrate fortified feeds and control diet on characteristics of salmon fillet. Thirty Atlantic salmon (6 per each group) were farmed for 12 weeks. All the fish were killed and frozen immediately. The white muscle from top posterior part of dorsal fin was dissected to analyze fat content, carotenoid content, content of water-soluble and salt-soluble proteins, cathepsin B and cathepsin B-L activities. ANOVA test was used to analyze mean and standard error of mean values at 0.05 significance level. There were significant difference in cathepsin B activity, water-soluble proteins and salt-soluble proteins (p-value= 0.005, 0.009 and 0.002). The mean values of other factors were not significantly different among the groups. Cathepsin B activity was higher in soymeal group. Water-soluble proteins were reported higher in soy meal and krill groups and salt-soluble proteins were significantly higher in soy meal and butyrate rich diets. Although soy meal has proven effect on enteritis, it results in higher percentage of protein in fillets. On the other hand, this feeding may have role in textural deterioration of fillets owing to higher values of endogenous cathepsin B in soymeal group.

Keywords: aquaculture, food quality, Krill protein extract, prebiotics, probiotics, Salmo salar, soy

Procedia PDF Downloads 194

Authors: Abeer Rababa'h, Ragad Bsoul, Mohammad Alkhatatbeh, Karem Alzoubi

Abstract:

Introduction: Tobacco use is one of the main risk factors to cardiovascular diseases (CVD) and atherosclerosis in particular. WPS contains several toxic materials such as: nicotine, carcinogens, tar, carbon monoxide and heavy metals. Thus, WPS is considered to be as one of the toxic environmental factors that should be investigated intensively. Therefore, the aim of this study is to investigate the effect of WPS on several cardiovascular biological markers that may cause atherosclerosis in mice. The study also conducted to study the temporal effects of WPS on the atherosclerotic biomarkers upon short (2 weeks) and long-term (8 weeks) exposures. Methods: mice were exposed to WPS and heart homogenates were analyzed to elucidate the effects of WPS on matrix metalloproteinase (MMPs), endothelin-1 (ET-1) and, myeloperoxidase (MPO). Following protein estimation, enzyme-linked immunosorbent assays were done to measure the levels of MMPs (isoforms 1, 3, and 9), MPO, and ET-1 protein expressions. Results: our data showed that acute exposure to WPS significantly enhances the levels of MMP-3, MMP- 9, and MPO expressions (p < 0.05) compared to their corresponding control. However, the body was capable to normalize the level of expressions for such parameters following continuous exposure for 8 weeks (p > 0.05). Additionally, we showed that the level of ET-1 expression was significantly higher upon chronic exposure to WPS compared to both control and acute exposure groups (p < 0.05). Conclusion: Waterpipe exposure has a significant negative effect on atherosclerosis and the enhancement of the atherosclerotic biomarkers expression (MMP-3 and 9, MPO, and ET-1) might represent an early scavenger of compensatory efforts to maintain cardiac function after WP exposure.

Keywords: atherosclerotic biomarkers, cardiovascular disease, matrix metalloproteinase, waterpipe

Procedia PDF Downloads 333

Authors: Mark-Adam Kellerman

Abstract:

Proteins are capable of exploring vast mutational spaces. This makes it difficult for protein engineers to devise rational methods to improve stability and function via mutagenesis. Deciding which residues to mutate requires knowledge of the characteristics they elicit. We probed the characteristics of residues in granulocyte-colony stimulating factor (G-CSF) using a thermal melt (from 295K to 323K) to denature it in a 700 MHz Bruker spectrometer. These characteristics included dynamics, micro-environmental changes experienced/ induced during denaturing and structure-function relationships. 15N-1H HSQC experiments were performed at 2K increments along with this thermal melt. We observed that dynamic residues that also undergo a lot of change in their microenvironment were predominantly in unstructured regions. Moreover, we were able to identify four residues (G4, A6, T133 and Q134) that we class as high priority targets for mutagenesis, given that they all appear in both the top 10% of measures for environmental changes and dynamics (∑Δ and ∆PI). We were also able to probe these NMR observables and combine them with molecular dynamics (MD) to elucidate what appears to be an opening motion of G-CSFs binding site III. V48 appears to be pivotal to this opening motion, which also seemingly distorts the loop region between helices A and B. This observation is in agreement with previous findings that the conformation of this loop region becomes altered in an aggregation-prone state of G-CSF. Hence, we present here an approach to profile the characteristics of residues in order to highlight their potential as rational mutagenesis targets and their roles in important conformational changes. These findings present not only an opportunity to effectively make biobetters, but also open up the possibility to further understand epistasis and machine learn residue behaviours.

Keywords: protein engineering, rational mutagenesis, NMR, molecular dynamics

Procedia PDF Downloads 235

Authors: Rumaisa Shahid, Saad Aziz Durrani, Shameel Pervez, Ibatsam Khokhar

Abstract:

Food waste is a prevalent issue in Pakistan, with over 40 percent of food discarded annually. Despite their decay, rotting fruits retain residual nutritional value consumed by microorganisms, notably fungi and bacteria. Fungi, preferred for their extracellular enzyme release, are gaining prominence, particularly for pectinase production. This enzyme offers several advantages, including clarifying juices by breaking down pectic compounds. In this study, three Aspergillus flavus isolates derived from decomposed fruits and manure were selected for pectinase production. The primary aim was to isolate fungi from diverse waste sources, identify the isolates and assess their capacity for pectinase production. The identification was done through morphological characteristics with the help of Light microscopy and Scanning Electron Microscopy (SEM). Pectinolytic potential was screened using pectin minimal salt agar (PMSA) medium, comparing clear zone diameters among isolates. Identification relied on morphological characteristics. Optimizing substrate (lemon and orange peel powder) concentrations, pH, temperature, and incubation period aimed to enhance pectinase yield. Spectrophotometry enabled quantitative analysis. The temperature was set at room temperature (28 ºC). The optimal conditions for Aspergillus flavus strain AF1(isolated from mango) included a pH of 5, an incubation period of 120 hours, and substrate concentrations of 3.3% for orange peels and 6.6% for lemon peels. For AF2 and AF3 (both isolated from soil), the ideal pH and incubation period were the same as AF1 i.e. pH 5 and 120 hours. However, their optimized substrate concentrations varied, with AF2 showing maximum activity at 3.3% for orange peels and 6.6% for lemon peels, while AF3 exhibited its peak activity at 6.6% for orange peels and 8.3% for lemon peels. Among the isolates, AF1 demonstrated superior performance under these conditions, comparatively.

Keywords: pectinase, lemon peel, orange peel, aspergillus flavus

Procedia PDF Downloads 55

Authors: Samira Khalaji, , Fenneke Klein Jan, Kay-E. Gottschalk, Eugenia Makrantonaki, Karin Scharffetter-Kochanek

Abstract:

Biological aging is a multi-dimensional process that takes place over a whole range of scales from the nanoscopic alterations within individual cells, over transformations in tissues and organs and to changes of the whole organism. On the single cell level, aging involves mutation of genes, differences in gene expression levels as well as altered posttranslational modifications of proteins. A variety of proteins is affected, including proteins of the cell cytoskeleton and migration machinery. Previous work quantified the expression of cytoskeleton proteins on the gene and protein levels in senescent and young fibroblasts. Their results show that senescent skin fibroblasts have an upregulated expression of the intermediate filament (IF) protein vimentin in contrast to actin and tubulin, which are downregulated. IFs play an important role in providing mechanical stability of cells. However, the mechanical properties of IFs depending on cellular senescence or age of the donor has not been studied so far. Hence, we employed passive microrheology on primary human dermal fibroblasts from female donors with age of 28 years (young) and 86 years (old) as model of in vivo aging and human normal dermal fibroblast from 11-year old male with CPD 17-35 (young) and CPD 58-59 (senescence) as a model of in vitro replicative senescence. In contrast to the expectations, our primary results show no significant differences in the viscoelastic properties of fibroblasts depending on age of the donor or cellular replicative senescence.

Keywords: aging, cytoskeleton, fibroblast, mechanical properties

Procedia PDF Downloads 296

Authors: Pornrut Rabintossaporn, Suphaket Saenthaweesuk, Amornnat Thuppia, Nuntiya Somparn

Abstract:

Several dietary and herbal plants have been shown to possess cytoprotective and antioxidant effects with various mechanisms of action. The aim of this study was to determine the antioxidant effects and its mechanism of aqueous leaves extract of Ocimum americanum (OA), commonly known as American basil or 'hoary basil', in rat. The extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with the extract at the dose of 100, 200 and 400 mg/kg for 28 days. Phytochemical screening of plant extracts revealed the presence of alkaloid, cardiac glycosides, tannin and steroid compounds. The extract contained phenolic compounds 36.91 ± 0.66 mg of gallic acid equivalents per gram OA extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 41.27 ± 1.86 µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14µg/mL. In the animals, the extract was well tolerated by the animals throughout the 28 days of study as shown by normal serum levels AST, ALP, ALT, BUN and Cr as well as normal histology of liver and pancreatic and kidney tissue. The protein expression of antioxidant enzymes, γ-glutamylcysteine ligase (γ-GCL) in liver was significantly increased compared with normal control. Consistent with the induction of γ-GCL protein expression significantly reduction of serum oxidative stress marker malondialdehyde (MDA) was found in rat treated with OA extract compared with control. Taken together, this study provides evidence that Ocimum americanum exhibits direct antioxidant properties and can induce cytoprotective enzyme in vivo.

Keywords: antioxidant, γ-glutamylcysteine ligase, MDA, Ocimum americanum

Procedia PDF Downloads 224

Authors: Arefeh Jafarian, Mohammad Taghikani, Saied Abroun, Amir Allahverdi, Masoud Soleimani

Abstract:

Introduction: The miRNAs have key roles in control of pancreatic islet development and insulin secretion. In this regards, current study investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. Findings: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose as well as extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. In derived IPCs by miR-375 alone are capable to express insulin and other endocrine specific transcription factors, the cells lack the machinery to respond to glucose. The differentiated hMSCs by miR-375 and anti-miR-9 lentiviruses could secrete insulin and c-peptide in a glucose-regulated manner. Conclusion: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus.

Keywords: diabetes, differentiation, MSCs, insulin producing cells, miR-375, miR-9

Procedia PDF Downloads 304

Authors: Vrushali Guhe, Shailza Singh

Abstract:

Cutaneous leishmaniasis is neglected tropical disease present worldwide caused by the protozoan parasite Leishmania major, the therapeutic armamentarium for leishmaniasis are showing several limitations as drugs are showing toxic effects with increasing resistance by a parasite. Thus identification of novel therapeutic targets is of paramount importance. Previous studies have shown that autophagy, a cellular process, can either facilitate infection or aid in the elimination of the parasite, depending on the specific parasite species and host background in leishmaniasis. In the present study, our objective was to target the essential autophagy protein ATG8, which plays a crucial role in the survival, infection dynamics, and differentiation of the Leishmania parasite. ATG8 in Leishmania major and its homologue, LC3, in Homo sapiens, act as autophagic markers. Present study manifested the crucial role of ATG8 protein as a potential target for combating Leishmania major infection. Through bioinformatics analysis, we identified non-conserved motifs within the ATG8 protein of Leishmania major, which are not present in LC3 of Homo sapiens. Against these two non-conserved motifs, we generated a peptide library of 60 peptides on the basis of physicochemical properties. These peptides underwent a filtering process based on various parameters, including feasibility of synthesis and purification, compatibility with Selective Reaction Monitoring (SRM)/Multiple reaction monitoring (MRM), hydrophobicity, hydropathy index, average molecular weight (Mw average), monoisotopic molecular weight (Mw monoisotopic), theoretical isoelectric point (pI), and half-life. Further filtering criterion shortlisted three peptides by using molecular docking and molecular dynamics simulations. The direct interaction between ATG8 and the shortlisted peptides was confirmed through Surface Plasmon Resonance (SPR) experiments. Notably, these peptides exhibited the remarkable ability to penetrate the parasite membrane and exert profound effects on Leishmania major. The treatment with these peptides significantly impacted parasite survival, leading to alterations in the cell cycle and morphology. Furthermore, the peptides were found to modulate autophagosome formation, particularly under starved conditions, suggesting their involvement in disrupting the regulation of autophagy within Leishmania major. In vitro, studies demonstrated that the selected peptides effectively reduced the parasite load within infected host cells. Encouragingly, these findings were corroborated by in vivo experiments, which showed a reduction in parasite burden upon peptide administration. Additionally, the peptides were observed to affect the levels of LC3II within host cells. In conclusion, our findings highlight the efficacy of these novel peptides in targeting Leishmania major’s ATG8 and disrupting parasite survival. These results provide valuable insights into the development of innovative therapeutic strategies against leishmaniasis via targeting autophagy protein ATG8 of Leishmania major.

Keywords: ATG8, leishmaniasis, surface plasmon resonance, MD simulation, molecular docking, peptide designing, therapeutics

Procedia PDF Downloads 62