Search results for: screening of diabetic retinopathy

Authors: Assia Belfar, Mohamed Hadjadj, Messaouda Dakmouche, Zineb Ghiaba

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Introduction: This study aims to phytochemical screening; Extracting the active compounds and estimate the effectiveness of antioxidant in Medicinal plants desert Limoniastrum guyonianum (Zeïta) from South Algeria. Methods: Total phenolic content and total flavonoid content using Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively. The total antioxidant capacity was estimated by the following methods: DPPH (1.1-diphenyl-2-picrylhydrazyl radical) and reducing power assay. Results: Phytochemical screening of the plant part reveals the presence of phenols, saponins, flavonoids and tannins. While alkaloids and Terpenoids were absent. The acetonic extract of L. guyonianum was extracted successively with ethyl acetate and butanol. Extraction of yield varied widely in the L. guyonianum ranging from (0.9425 %to 11.131%). The total phenolic content ranged from 53.33 mg GAE/g DW to 672.79 mg GAE/g DW. The total flavonoid concentrations varied from 5.45 to 21.71 mg/100g. IC50 values ranged from 0.02 ± 0.0004 to 0.13 ± 0.002 mg/ml. All extracts showed very good activity of ferric reducing power, the higher power was in butanol fraction (23.91 mM) more effective than BHA, BHT and VC. Conclusions: Demonstrated this study that the acetonic extract of L. guyonianum contain a considerable quantity of phenolic compounds and possess a good antioxidant activity. Can be used as an easily accessible source of Natural Antioxidants and as a possible food supplement and in the pharmaceutical industry.

Keywords: limoniastrum guyonianum, phenolics compounds, flavonoid compound, antioxidant activity

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Authors: Nidhi Chadha, A. K. Tiwari, Marilyn D. Milton, Anil K. Mishra

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Translocator protein (18 kDa) TSPO is highly expressed during microglia activation in neuroinflammation. Although a number of PET ligands have been developed for the visualization of activated microglia, one of the advantageous approaches is to develop potential optical imaging (OI) probe. Our study involves computational screening, synthesis and evaluation of TSPO ligand through various imaging modalities namely PET/SPECT/Optical. The initial computational screening involves pharmacophore modeling from the library designing having oxo-benzooxazol-3-yl-N-phenyl-acetamide groups and synthesis for visualization of efficacy of these compounds as multimodal imaging probes. Structure modeling of monomer, Ala147Thr mutated, parallel and anti-parallel TSPO dimers was performed and docking analysis was performed for distinct binding sites. Computational analysis showed pattern of variable binding profile of known diagnostic ligands and NBMP via interactions with conserved residues along with TSPO’s natural polymorphism of Ala147→Thr, which showed alteration in the binding affinity due to considerable changes in tertiary structure. Preliminary in vitro binding studies shows binding affinity in the range of 1-5 nm and selectivity was also certified by blocking studies. In summary, this skeleton was found to be potential probe for TSPO imaging due to ease in synthesis, appropriate lipophilicity and reach to specific region of brain.

Keywords: TSPO, molecular modeling, imaging, docking

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Authors: Li-Ching Lin, Yu-Tzu Dai

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Recurrent foot ulcers or foot amputation have a major impact on patients with diabetes mellitus (DM), medical professionals, and society. A critical procedure for foot care is foot self-monitoring. Medical professionals’ understanding of patients’ foot self-monitoring knowledge, attitude, and practice is beneficial for raising patients’ disease awareness. This study investigated these and related factors among patients with DM through a descriptive study of the correlations. A scale for measuring the foot self-monitoring knowledge, attitude, and practice of patients with DM was used. Purposive sampling was adopted, and 100 samples were collected from the respondents’ self-reports or from interviews. The statistical methods employed were an independent-sample t-test, one-way analysis of variance, Pearson correlation coefficient, and multivariate regression analysis. The findings were as follows: the respondents scored an average of 12.97 on foot self-monitoring knowledge, and the correct answer rate was 68.26%. The respondents performed relatively lower in foot health screenings and recording, and awareness of neuropathy in the foot. The respondents held a positive attitude toward self-monitoring their feet and a negative attitude toward having others check the soles of their feet. The respondents scored an average of 12.64 on foot self-monitoring practice. Their scores were lower in their frequency of self-monitoring their feet, recording their self-monitoring results, checking their pedal pulse, and examining if their soles were red immediately after taking off their shoes. Significant positive correlations were observed among foot self-monitoring knowledge, attitude, and practice. The correlation coefficient between self-monitoring knowledge and self-monitoring practice was 0.20, and that between self-monitoring attitude and self-monitoring practice was 0.44. Stepwise regression analysis revealed that the main predictive factors of the foot self-monitoring practice in patients with DM were foot self-monitoring attitude, prior experience in foot care, and an educational attainment of college or higher. These factors predicted 33% of the variance. This study concludes that patients with DM lacked foot self-monitoring practice and advises that the patients’ self-monitoring abilities be evaluated first, including whether patients have poor eyesight, difficulties in bending forward due to obesity, and people who can assist them in self-monitoring. In addition, patient education should emphasize self-monitoring knowledge and practice, such as perceptions regarding the symptoms of foot neurovascular lesions, pulse monitoring methods, and new foot self-monitoring equipment. By doing so, new or recurring ulcers may be discovered in their early stages.

Keywords: diabetic foot, foot self-monitoring attitude, foot self-monitoring knowledge, foot self-monitoring practice

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Authors: Dara Kutsyk, Olga Bondarenko, Mariya Sorochka

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In 21-century type 2 diabetes (T2D) has become a global health and social problem in the whole world. The goal of treatment for patients with T2D is to prevent complications of diabetes - macrovascular diseases (heart disease, stroke, and peripheral vascular disease) and microvascular diseases (retinopathy, neuropathy and nephropathy). Nitric oxide (NO) plays an important role in maintaining vascular homeostasis. Loss of NO function is one of the earliest indicators of disease and its progression especially in patients with T2D. Aim: To compare NO level between patients with well and bad controlled glycemia in T2D. Methods: The study included 32 patients with T2D. The diagnosis of T2D was confirmed due to International Diabetes Federation (IDF) criteria 2015. Patients were divided into two groups: with well controlled glycaemia (HbA1c < 7%) and bad controlled glycaemia (HbA1c > 7%). The control group consists of 15 healthy subjects. Results: NO level in patients with T2D is significantly higher (27,2 ±3,1 µmol), compared to controls (18,86±0,9 µmol; p < 0,001). A significant difference in NO level was found between patients with bad controlled glycaemia (25,9±2,2 µmol) and well controlled glycaemia (28,7 ± 3,0 µmol; p<0,01). The study showed a moderate negative correlation between NO level and HbA1c (-0,399; р< 0,05). Conclusions: Production of NO is impaired in patients with T2D, especially with badly controlled glycaemia. With the increase in HbAc serum NO decreases. This can be the main target for prevention vascular complication in T2D.

Keywords: type 2 diabetes, glycated hemoglobin, nitric oxide, Diabetes mellitus

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Authors: Shahid Mahboob, Hafiz Muhammad Ashraf, Salma Sultana, Tayyaba Sultana, Khalid Al-Ghanim, Fahid Al-Misned, Zubair Ahmedd

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There is an increasing need for an effective tool to estimate the risks derived from the large number of pollutants released to the environment by human activities. Typical screening procedures are highly invasive or lethal to the fish. Recent studies show that fish scales biochemically respond to a range of contaminants, including toxic metals, organic compounds, and endocrine disruptors. The present study evaluated the effects of the surface water contaminants on Cyprinus carpio in the Ravi River by comparing DNA extracted non-lethally from their scales to DNA extracted from the scales of fish collected from a controlled fish farm. A single, random sampling was conducted. Fish were broadly categorised into three weight categories (W1, W2 and W3). The experimental samples in the W1, W2 and W3 categories had an average DNA concentration (µg/µl) that was lower than the control samples. All control samples had a single DNA band; whereas the experimental samples in W1 fish had 1 to 2 bands, the experimental samples in W2 fish had two bands and the experimental samples in W3 fish had fragmentation in the form of three bands. These bands exhibit the effects of pollution on fish in the Ravi River. On the basis findings of this study, we propose that fish scales can be successfully employed as a new non-lethal tool for the evaluation of the effect of surface water contaminants.

Keywords: fish scales, Cyprinus carpio, heavy metals, non-invasive, DNA fragmentation

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Authors: Jarel Elgin Tolentino, Arby Denise Nera, Mary Rose Roco, Angela Vianca Aspa, Nikko Beltran, Else Dapat

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Drug resistance by cells has been the problem in the medical field for decades now. The use of medicinal plants as a source of creating powerful drugs has been nowadays recognized worldwide to treat such resistant diseases. In the present study, the potential for Diospyros philippinensis A. DC. to inhibit growth of both bacteria and cancer cell line was conducted. The leaf crude extracts were screened for the presence of phytochemicals and examined for potential bioactivities by employing several assays like Kirby-Bauer disc diffusion method, DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium assay for the antibacterial, antioxidant and cytotoxic activities of the extract, respectively. Phytochemical test results of the extracts revealed the presence of alkaloids, flavonoids, saponins, phenols, quinones, cardiac glycosides, phlobatannins, carbohydrate, cardenolides and proteins. The leaf extracts were found to exhibit antibacterial activity against gram-positive bacteria, high antioxidant activity (99.22% ± 0.005) but did not show any sign of cytotoxicity towards HCT116 (ATCC CCL-247). The study therefore concludes that D. philippinensis A. DC. leaf extract can be a source of antibacterial and chemopreventive agents. This claim may be used as basis for future investigation.

Keywords: bioassay, medicinal plants, plant crude extracts, phytochemical screening

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Authors: Mehwish Asghar

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Breast Cancer (BC) is a common type of cancer among women. Its screening is usually performed using different imaging modalities such as magnetic resonance imaging, mammogram, X-ray, CT, etc. Among these modalities’ mammogram is considered a powerful tool for diagnosis and screening of breast cancer. Sophisticated machine learning approaches have shown promising results in complementing human diagnosis. Generally, machine learning methods can be divided into two major classes: one is Radiomics analysis (RA), where image features are extracted manually; and the other one is the concept of convolutional neural networks (CNN), in which the computer learns to recognize image features on its own. This research aims to improve the incidence of early detection, thus reducing the mortality rate caused by breast cancer through the latest advancements in computer science, in general, and machine learning, in particular. It has also been aimed to ease the burden of doctors by improving and automating the process of breast cancer detection. This research is related to a relative analysis of different techniques for the implementation of different models for detecting and classifying breast cancer. The main goal of this research is to provide a detailed view of results and performances between different techniques. The purpose of this paper is to explore the potential of a convolutional neural network (CNN) w.r.t feature extractor and as a classifier. Also, in this research, it has been aimed to add the module of Radiomics for comparison of its results with deep learning techniques.

Keywords: breast cancer (BC), machine learning (ML), convolutional neural network (CNN), radionics, magnetic resonance imaging, artificial intelligence

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Authors: Anupalli Roja Rani, Saraswathi Jaggali

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Rose-scented geranium (Pelargonium graveolens L’Hér.) is an erect, much-branched shrub. It is indigenous to various parts of southern Africa, and it is often called Geranium. Pelargonium species are widely used by traditional healers in the areas of Southern Africa by Sotho, Xhosa, Khoi-San and Zulus for its curative and palliative effects in the treatment of diarrhea, dysentery, fever, respiratory tract infections, liver complaints, wounds, gastroenteritis, haemorrhage, kidney and bladder disorders. We have used Plant materials for extracting active compounds from analytical grades of solvents methanol, ethyl acetate, chloroform and water by a soxhlet apparatus. The phytochemical screening reveals that extracts of Pelargonium graveolens contains alkaloids, glycosides, steroids, tannins, saponins and phenols in ethyl acetate solvent. The antioxidant activity was determined using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) bleaching method and the total phenolic content in the extracts was determined by the Folin–Ciocalteu method. Due to the presence of different phytochemical compounds in Pelargonium the anti-microbial activity against different micro-organisms like E.coli, Streptococcus, Klebsiella and Bacillus. Fractionation of plant extract was performed by column chromatography and was confirmed with HPLC analysis, NMR and FTIR spectroscopy for the compound identification in different organic solvent extracts.

Keywords: Pelargonium graveolens L’Hér, DPPH, micro-organisms, HPLC analysis, NMR, FTIR spectroscopy

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Authors: Geetha Jayapathy, Lakshmi Muthukrishnan, Manoj Kumar Reddy Pulim , Radhika Raman

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Introduction: Ketones are the end products of fatty acid metabolism and a source of energy for vital organs such as the brain, heart and skeletal muscles. Ketones are produced in excess when glucose is not available as a source of energy or it cannot be utilized as in diabetic ketoacidosis. Children admitted in the emergency department often have starvation ketosis which is not clinically manifested. Decision on admission of children to the emergency room with subtle signs can be difficult at times. Point of care blood ketone testing can be done at the bedside even in a primary level care setting to supplement and guide us in our management decisions. Hence this study was done to explore the utility of this simple bedside parameter as a supplement in assessing pediatric patients presenting to the emergency department. Objectives: To estimate blood ketones of children admitted in the emergency department. To analyze the significance of blood ketones in various disease conditions. Methods: Blood ketones using point of care testing instrument (ABOTTprecision Xceed Pro meters) was done in patients getting admitted in emergency room and in out-patients (through sample collection centre). Study population: Children aged 1 month to 18 years were included in the study. 250 cases (In-patients) and 250 controls (out-patients) were collected. Study design: Prospective observational study. Data on details of illness and physiological status were documented. Blood ketones were compared between the two groups and all in patients were categorized into various system groups and analysed. Results: Mean blood ketones were high in in-patients ranging from 0 to 7.2, with a mean of 1.28 compared to out-patients ranging from 0 to 1.9 with a mean of 0.35. This difference was statistically significant with a p value < 0.001. In-patients with shock (mean of 4.15) and diarrheal dehydration (mean of 1.85) had a significantly higher blood ketone values compared to patients with other system involvement. Conclusion: Blood ketones were significantly high (above the normal range) in pediatric patients who are sick requiring admission. Patients with various forms of shock had very high blood ketone values as found in diabetic ketoacidosis. Ketone values in diarrheal dehydration were moderately high correlating to the degree of dehydration.

Keywords: admission, blood ketones, paediatric emergencies, point of care testing

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Authors: Lynette Lincoln, Sunil S. More

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With the rising demand of sugar used today, it is proposed that world sugar is expected to escalate up to 203 million tonnes by 2021. Hydrolysis of sucrose (table sugar) into glucose and fructose equimolar mixture is catalyzed by β-D-fructofuranoside fructohydrolase (EC 3.2.1.26), commonly called as invertase. For fluid filled center in chocolates, preparation of artificial honey, as a sweetener and especially to ensure that food stuffs remain fresh, moist and soft for longer spans invertase is applied widely and is extensively being used. From an industrial perspective, properties such as increased solubility, osmotic pressure and prevention of crystallization of sugar in food products are highly desired. Screening for invertase does not involve plate assay/qualitative test to determine the enzyme production. In this study, we use a three-step screening strategy for identification of a novel bacterial isolate from soil which is positive for invertase production. The primary step was serial dilution of soil collected from sugarcane fields (black soil, Maddur region of Mandya district, Karnataka, India) was grown on a Czapek-Dox medium (pH 5.0) containing sucrose as the sole C-source. Only colonies with the capability to utilize/breakdown sucrose exhibited growth. Bacterial isolates released invertase in order to take up sucrose, splitting the disaccharide into simple sugars. Secondly, invertase activity was determined from cell free extract by measuring the glucose released in the medium at 540 nm. Morphological observation of the most potent bacteria was examined by several identification tests using Bergey’s manual, which enabled us to know the genus of the isolate to be Bacillus. Furthermore, this potent bacterial colony was subjected to 16S rDNA PCR amplification and a single discrete PCR amplicon band of 1500 bp was observed. The 16S rDNA sequence was used to carry out BLAST alignment search tool of NCBI Genbank database to obtain maximum identity score of sequence. Molecular sequencing and identification was performed by Xcelris Labs Ltd. (Ahmedabad, India). The colony was identified as Bacillus sp. BAB-3434, indicating to be the first novel strain for extracellular invertase production. Molasses, a by-product of the sugarcane industry is a dark viscous liquid obtained upon crystallization of sugar. An enhanced invertase production and optimization studies were carried out by one-factor-at-a-time approach. Crucial parameters such as time course (24 h), pH (6.0), temperature (45 °C), inoculum size (2% v/v), N-source (yeast extract, 0.2% w/v) and C-source (molasses, 4% v/v) were found to be optimum demonstrating an increased yield. The findings of this study reveal a simple screening method of an extracellular invertase from a rapidly growing Bacillus sp., and selection of best factors that elevate enzyme activity especially utilization of molasses which served as an ideal substrate and also as C-source, results in a cost-effective production under submerged conditions. The invert mixture could be a replacement for table sugar which is an economic advantage and reduce the tedious work of sugar growers. On-going studies involve purification of extracellular invertase and determination of transfructosylating activity as at high concentration of sucrose, invertase produces fructooligosaccharides (FOS) which possesses probiotic properties.

Keywords: Bacillus sp., invertase, molasses, screening, submerged fermentation

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Authors: K. S. Al-Mugren, A. Yahya, S. Alodah, R. Alharbi, S. H. Almsaid , A. Alqahtani, H. Jaber, A. Basaqer, N. Alajra, N. Almoghati, A. Alsalman, Khalid Alharbi

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Context: The production of biofertilizers from naturally occurring microorganisms is an area of research that aims to enhance agricultural practices by utilizing local resources. This research project focuses on isolating and screening indigenous microorganisms with PK-fixing and phosphate solubilizing characteristics from local sources. Research Aim: The aim of this project is to develop a biofertilizer product using indigenous microorganisms and composted agro waste as a carrier. The objective is to enhance crop productivity and soil fertility through the application of biofertilizers. Methodology: The research methodology includes several key steps. Firstly, indigenous microorganisms will be isolated from local resources using the ten-fold serial dilutions technique. Screening assays will be conducted to identify microorganisms with phosphate solubilizing and PK-fixing activities. Agro-waste materials will be collected from local agricultural sources, and composting experiments will be conducted to convert them into organic matter-rich compost. Physicochemical analysis will be performed to assess the composition of the composted agro-waste. Gamma and X-ray irradiation will be used to sterilize the carrier material. The sterilized carrier will be tested for sterility using the ten-fold serial dilutions technique. Finally, selected indigenous microorganisms will be developed into biofertilizer products. Findings: The research aims to find suitable indigenous microorganisms with phosphate solubilizing and PK-fixing characteristics for biofertilizer production. Additionally, the research aims to assess the suitability of composted agro waste as a carrier for biofertilizers. The impact of gamma irradiation sterilization on pathogen elimination will also be investigated. Theoretical Importance: This research contributes to the understanding of utilizing indigenous microorganisms and composted agro waste for biofertilizer production. It expands knowledge on the potential benefits of biofertilizers in enhancing crop productivity and soil fertility. Data Collection and Analysis Procedures: The data collection process involves isolating indigenous microorganisms, conducting screening assays, collecting and composting agro waste, analyzing the physicochemical composition of composted agro waste, and testing carrier sterilization. The analysis procedures include assessing the abilities of indigenous microorganisms, evaluating the composition of composted agro waste, and determining the sterility of the carrier material. Conclusion: The research project aims to develop biofertilizer products using indigenous microorganisms and composted agro waste as a carrier. Through the isolation and screening of indigenous microorganisms, the project aims to enhance crop productivity and soil fertility by utilizing local resources. The research findings will contribute to the understanding of the suitability of composted agro waste as a carrier and the efficacy of gamma irradiation sterilization. The research outcomes will have theoretical importance in the field of biofertilizer production and agricultural practices.

Keywords: biofertilizer, microorganisms, agro waste, nuclear technologies

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Authors: Azizollah Khodakaram- Tafti, Ali Mohammadi, Ghasem Farjanikish

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Bovine viral diarrhea virus (BVDV) is one of the most important viral pathogens of cattle worldwide caused by Pestivirus genus, Flaviviridae family.The aim of the present study was to comparison several diagnostic methods and determine the prevalence of BVDV infection for the first time in dairy herds of Fars province, Iran. For initial screening, a total of 400 blood samples were randomly collected from 12 industrial dairy herds and analyzed using reverse transcription (RT)-PCR on the buffy coat. In the second step, blood samples and also ear notch biopsies were collected from 100 cattle of infected farms and tested by antigen capture ELISA (ACE), RT-PCR and immunohistochemistry (IHC). The results of nested RT-PCR (outer primers 0I100/1400R and inner primers BD1/BD2) was successful in 16 out of 400 buffy coat samples (4%) as acute infection in initial screening. Also, 8 out of 100 samples (2%) were positive as persistent infection (PI) by all of the diagnostic tests similarly including RT-PCR, ACE and IHC on buffy coat, serum and skin samples, respectively. Immunoreactivity for bovine BVDV antigen as brown, coarsely to finely granular was observed within the cytoplasm of epithelial cells of epidermis and hair follicles and also subcutaneous stromal cells. These findings confirm the importance of monitoring BVDV infection in cattle of this region and suggest detection and elimination of PI calves for controlling and eradication of this disease.

Keywords: antigen capture ELISA, bovine viral diarrhea virus, immunohistochemistry, RT-PCR, cattle

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Authors: Amal Saafan, Kareem Al Sofy, Sameh AbdelGhani, Magdy Amin

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Background: Acinetobacter spp. resistance towards β-lactam antibiotics is mediated mainly by different classes of β-lactamases production; detection of some genes responsible for production of β-lactamases is the objective of the study. Methods: One hundred fifty bacterial isolates were recovered from blood, sputum, and urine specimens from different hospitals in Egypt. Sixty-nine isolate were identified as Acinetobacter baumannii using traditional biochemical tests, CHROM agar, MicroScan and PCR amplification of blaoxa-51like gene. Acinetobacterbaumannii isolates were grouped into carbapenem resistant group (GP1), cefotaxime, ceftazidime and cefoxitin resistant group (GP2) and carbapenem and cephalosporin non-resistant group (GP3). Carbapenemase activity was screened using modified Hodge test (MHT) for GP1.Metallo-β-lactamases screening was performed for MHT positive isolates using double disk synergy test (DDST) and combined disk test (CDT). Amp C activity was screened using Amp C disk test with Tris-EDTA, DDST, and CDT for GP2. Finally, PCR amplification of blaoxa-51like, blaoxa-23like, blaIMP-like, blaVIM-like, and blaADC-like genes was performed for isolates that showed, at least, two positive results of three for both AmpC and carbapenemases phenotypic screening tests (obvious activity), in addition to GP3 (for comparison). Detection of blaoxa-51like and blaADC-like genes preceded by ISAba1 was also performed. Results: Antibiogram of 69 pure Acinetobacter baumannii isolates resulted in 57, 64, and 2 isolates enrolled into GP1, GP2, and GP3, respectively. Carbapenemase activity was shown by 49(85.9%) isolate using MHT. Metallo-β-lactamases screening revealed 32(65.3%) and 35(71.4%) using DDST and CDT, respectively.AmpC activity was shown by 43(67.2%) and 50 (78.1%) isolates using AmpC disk test with Tris-EDTA, and both DDST and CDT, respectively. Twenty-seven isolates showed obvious activity, all of them (100%) were harboring blaoxa-51like and blaADC-like genes, while blaoxa-23like, blaIMP-like andblaVIM-like genes were harbored by 23(85.2%), 9 (33.%) and no isolate respectively. Only 12 (44.4%) isolates harbored blaoxa-51like and blaADC-like genes preceded by ISAba1. GP3 isolates showed only positive blaoxa-51like and blaADC-like genes. Conclusion: It is not possible to correlate resistance with presence of blaoxa-51like and blaADC-like genes and presence of ISAba1 was immediate as transcriptional promoter. A blaoxa-23like gene played an important role in carbapenem resistance when compared with blaIMP-like and blaVIM-like gene.

Keywords: acinetobacter, beta-lactams, resistance, antimicrobial agents

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Authors: Amer A. Hasan, Mehri Igci, Ersin Borazan, Rozhgar A. Khailany, Emine Bayraktar, Ahmet Arslan

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Gastric cancer (GC) has high morbidity and fatality rate in various countries and is still one of the most frequent and deadly diseases. Novel mitogenic and motogenic Gastrokine1 (GKN1) and Gastrokine 2 (GKN2) genes that are highly expressed in the normal stomach epithelium and plays an important role in maintaining the integrity and homeostasis of stomach mucosal epithelial cells. Significant loss of copy number and mRNA transcript of GKN1 and GKN2 gene expression were frequently observed in all types of gastric cancer. In this study, 47 paired samples that were grouped according to the types of gastric cancer and the clinical characteristics of the patients, including gender and average of age were investigated with gene expression analysis and mutation screening by monetering RT-PCR, SSCP and nucleotide sequencing techniques. Both GKN1 and GKN2 genes were observed significantly reduced found by (Wilcoxon signed rank test; p<0.05). As a result of gene screening, no mutation (no different genotype) was detected. It is considered that gene mutations are not the cause of inactivation of gastrokines. In conclusion, the mRNA expression level of GKN1 and GKN2 genes statistically was decreased regardless the gender, age or cancer type of patients. Reduced of gastrokine genes seems to occur at the initial steps of cancer development. In order to understand the investigation between gastric cancer and diagnostic biomarker; further analysis is necessary.

Keywords: gastric cancer, diagnostic biomarker, nucleotide sequencing, semi-quantitative RT-PCR

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Authors: Daniel Wachira

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The study aims to determine factors contributing to increasing rates of DFU among DM patients attending clinics in Nyahururu County referral hospital, Lakipia County. The study objectives include;- To determine the demographic factors contributing to increased rates of DFU among DM patients, determining the sociocultural factors that contribute to increased rates of DFU among DM patients and determining the health facility factors contributing to increased rates of DFU among DM patients attending DM clinic at Nyahururu county referral hospital, Laikipia County. This study will adopt a descriptive cross-sectional study design. It involves the collection of data at a one-time point without follow-up. This method is fast and inexpensive, there is no loss to follow up as the data is collected at one time point and associations between variables can be determined. The study population includes all DM patients with or without DFU. The sampling technique that will be used is the probability sampling method, a simple random method of sampling will be used. The study will employ the use of questionnaires to collect the required information. Questionnaires will be a research administered questionnaires. The questionnaire developed was done in consultation with other research experts (supervisor) to ensure reliability. The questionnaire designed will be pre-tested by hand delivering them to a sample 10% of the sample size at J.M Kariuki Memorial hospital, Nyandarua county and thereafter collecting them dully filled followed by refining of errors to ensure it is valid for collection of data relevant for this study. Refining of errors on the questionnaires to ensure it was valid for collection of data relevant for this study. Data collection will begin after the approval of the project. Questionnaires will be administered only to the participants who met the selection criteria by the researcher and those who agreed to participate in the study to collect key information with regard to the objectives of the study. The study's authority will be obtained from the National Commission of Science and Technology and Innovation. Permission will also be obtained from the Nyahururu County referral hospital administration staff. The purpose of the study will be explained to the respondents in order to secure informed consent, and no names will be written on the questionnaires. All the information will be treated with maximum confidentiality by not disclosing who the respondent was and the information.

Keywords: diabetes, foot ulcer, social factors, hospital factors

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Authors: Anjani Kumar Tiwari, Neelam Kumari, Anil Mishra

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The 18-kDa translocator protein (TSPO) previously called as peripheral benzodiazepine receptor, is proven biomarker for variety of neuroinflammation. TSPO is tryptophane rich five transmembranal protein found on outer mitochondrial membrane of steroid synthesising and immunomodulatory cells. In case of neuronal damage or inflammation the expression level of TSPO get upregulated as an immunomodulatory response. By utilizing Benzoxazolone as a basic scaffold, series of TSPO ligands have been designed followed by their screening through in silico studies. Synthesis has been planned by employing convergent methodology in six high yielding steps. For the synthesized ligands the ‘in vitro’ assay was performed to determine the binding affinity in term of Ki. On ischemic rat brain, autoradiography studies were also carried to check the specificity and affinity of the designed radiolabelled ligand for TSPO.Screening was performed on the basis of GScore of CADD based schrodinger software. All the modified and better prospective compound were successfully carried out and characterized by spectroscopic techniques (FTIR, NMR and HRMS). In vitro binding assay showed best binding affinity Ki = 6.1+ 0.3 for TSPO over central benzodiazepine receptor (CBR) Ki > 200. ARG studies indicated higher uptake of two analogues on the lesion side compared with that on the non-lesion side of ischemic rat brains. Displacement experiments with unlabelled ligand had minimized the difference in uptake between the two sides which indicates the specificity of the ligand towards TSPO receptor.

Keywords: TSPO, PET, imaging, Acetamidobenzoxazolone

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Authors: J. K. D. M. P. Madara, R. B. L. Dharmawickreme, Linu John, Ivee Boiss

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Flemingia vestita, commonly known as ‘Sohphlang’ is an important medicinal plant found in the North-Eastern region of India, which is traditionally recognized for its anthelmintic properties. This study was aimed to evaluate the phytochemical constituents and antibacterial activity of the tuber skin extracts of the plant species. Methanol, acetone, and water were used to obtain the solvent extractions of the skin peel extracts. Concentrated extracts of skin peel were tested using previously established qualitative phytochemical assays. The antibacterial efficacy of methanol tuber skin extract was tested against Gram-negative and positive microorganisms, namely, Klebsiella pneumonia, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Mycobacterium tuberculosis strains. Agar well diffusion method was employed to determine the zone of inhibition of the plant extracts. Obtained data were statistically analyzed. Methanol extracts of Flemingia vestita were found to be effective against Bacillus subtilis and Mycobacterium tuberculosis at concentrations of 0.5 mg/ml. The reported zone of inhibition for the two strains was 13.3mm ± 0.57 and 16.3mm ± 4.9, respectively. However Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli were resistant to the plant extracts with no zone of inhibition. Alkaloids, glycosides, and phenols were found to be present in aqueous, methanol, and acetone extracts of the plant in qualitative phytochemical analysis.

Keywords: flemingia vestita, antibacterial activity, phytochemical screening, well diffusion method

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Authors: Gabriel S. De Oliveira, Patricia P. Adriani, Christophe Moriseau, Bruce D. Hammock, Felipe S. Chambergo

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Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have high biotechnological interest for the drug design and chemistry transformation for industries. In this study, we describe the identification of substrates and inhibitors of epoxide hydrolase enzyme from the filamentous fungus Trichoderma reesei (TrEH), and these inhibitors showed the fungal growth inhibitory activity. We have used the cloned enzyme and expressed in E. coli to develop the screening in the library of fluorescent substrates with the objective of finding the best substrate to be used in the identification of good inhibitors for the enzyme TrEH. The substrate (3-phenyloxiranyl)-acetic acid cyano-(6-methoxy-naphthalen-2-yl)-methyl ester showed the highest specific activity and was chosen for the next steps of the study. The inhibitors screening was performed in the library with more than three thousand molecules and we could identify the 6 best inhibitors. The IC50 of these molecules were determined in nM and all the best inhibitors have urea or amide in their structure, because It has been recognized that these groups fit well in the hydrolase catalytic pocket of the epoxide hydrolases. Then the growth of T. reesei in PDA medium containing these TrEH inhibitors was tested, and fungal growth inhibition activity was demonstrated with more than 60% of inhibition of fungus growth in the assay with the TrEH inhibitor with the lowest IC50. Understanding how this EH enzyme from T. reesei responds to inhibitors may contribute for the study of fungal metabolism and drug design against pathogenic fungi.

Keywords: epoxide hydrolases, fungal growth inhibition, inhibitor, Trichoderma reesei

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Authors: Aisling Eves, Andrew Pieri, Ross McLean, Nerys Forester

Abstract:

Background: The breast unit local guideline recommends that 12 cores should be used in a stereotactic-guided biopsy to diagnose DCIS. Twelve cores are regarded to provide good diagnostic value without removing more breast tissue than necessary. This study aimed to determine compliance with guidelines and investigated how the number of cores impacted upon the re-excision rate and size discrepancies. Methods: This single-centre retrospective cohort study of 72 consecutive breast screened patients with <15mm DCIS on radiological report underwent stereotactic-guided core biopsy and subsequent surgical excision. Clinical, radiological, and histological data were collected over 5 years, and ASCO guidelines for margin involvement of <2mm was used to guide the need for re-excision. Results: Forty-six (63.9%) patients had <12 cores taken, and 26 (36.1%) patients had ≥12 cores taken. Only six (8.3%) patients had 12 cores taken in their stereotactic biopsy. Incomplete surgical excision was seen in 17 patients overall (23.6%), and of these patients, twelve (70.6%) had fewer than 12 cores taken (p=0.55 for the difference between groups). Mammogram and biopsy underestimated the size of the DCIS in this subgroup by a median of 15mm (range: 6-135mm). Re-excision was required in 9 patients (12.5%), and five patients (6.9%) were found to have invasive ductal carcinoma on excision (80% had <12 cores, p=0.43). Discussion: There is poor compliance with the breast unit local guidelines and higher rates of re-excision in patients who did not have ≥12 cores taken. Taking ≥12 cores resulted in fewer missed invasive cancers lower incomplete excision and re-excision rates.

Keywords: stereotactic core biopsy, DCIS, breast screening, Re-excision rates, core biopsy

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Authors: Restuning Widiasih, Katherine Nelson, Joan Skinner

Abstract:

Muslim husbands have significant roles in the family including their roles in women’s health and illness. However, studies that explore Muslim husbands’ participation in women’s health is limited. The objective of this study was to uncover Muslim husbands’ participation in women’ health and illness including cancer prevention and screening. A descriptive exploratory approach was used involving 20 Muslim women from urban and rural areas of West Java Province, Indonesia. Muslim women shared experience related to their husbands support and activities in women’s health and illness. The data from the interviews were analyzed using the Comparative Analysis for Interview (CAI). Women perceived that husbands fully supported their health by providing opportunities for activities, and reminding them about healthy food, their workloads, and family planning. Husbands actively involved when women faced health issues including sharing knowledge and experience, discussing any health problems, advising for medical check-ups, and accompanying them for treatments. The analysis also found that husbands were less active and offered less advice regarding prevention and early detection of cancer. This study highlights the significant involvement of Muslim husbands in women’s health and illness, yet a lack of support from husbands related to screening and cancer prevention. This condition could be a burden for Muslim women to participate in health programs related to cancer prevention and early detection. Health education programs to improve Muslim husbands’ understanding of women’s health is needed.

Keywords: descriptive exploratory study, Muslim husbands, Muslim women, women's health and illness

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Authors: Prof. Ammal Mokhtar Metwally, Samia M. Sami, Nihad A. Ibrahim, Fatma A. Shaaban, Iman I. Salama

Abstract:

Objectives: Reducing neonatal mortality by 2030 is still a challenging goal in developing countries. low birth weight (LBW) is a significant contributor to this, especially where weighing newborns is not possible routinely. The present study aimed to determine a simple, easy, reliable anthropometric measure(s) that can predict LBW) and neonatal mortality. Methods: A prospective cohort study of 570 babies born in districts of El Menia governorate, Egypt (where most deliveries occurred at home) was examined at birth. Newborn weight, length, head, chest, mid-arm, and thigh circumferences were measured. Follow up of the examined neonates took place during their first four weeks of life to report any mortalities. The most predictable anthropometric measures were determined using the statistical package of SPSS, and multiple Logistic regression analysis was performed.: Results: Head and chest circumferences with cut-off points < 33 cm and ≤ 31.5 cm, respectively, were the significant predictors for LBW. They carried the best combination of having the highest sensitivity (89.8 % & 86.4 %) and least false negative predictive value (1.4 % & 1.7 %). Chest circumference with a cut-off point ≤ 31.5 cm was the significant predictor for neonatal mortality with 83.3 % sensitivity and 0.43 % false negative predictive value. Conclusion: Using chest circumference with a cut-off point ≤ 31.5 cm is recommended as a single simple anthropometric measurement for the prediction of both LBW and neonatal mortality. The predicted measure could act as a substitute for weighting newborns in communities where scales to weigh them are not routinely available.

Keywords: low birth weight, neonatal mortality, anthropometric measures, practical screening

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Authors: Georges Jabbour

Abstract:

Background and aims: This two-year follow-up study explores the long-term sustainability of physical activity (PA) levels in young people with type 1 diabetes, focusing on the relationship between PA, hypoglycemia, and behavioral scores. The literature highlights the importance of PA and its health benefits, as well as the barriers to engaging in PA practices. Studies have shown that individuals with high levels of vigorous physical activity have higher fear of hypoglycemia (FOH) scores and more hypoglycemia episodes. Considering that hypoglycemia episodes are a major barrier to physical activity, and many studies reported a negative association between PA and high FOH scores, it cannot be guaranteed that those experiencing hypoglycemia over a long period will remain active. Building on that, the present work assesses whether high PA levels, despite elevated hypoglycemia risk, can be maintained over time. The study tracks PA levels at one and two years, correlating them with hypoglycemia instances and Fear of Hypoglycemia (FOH) scores. Materials and methods: A self-administered questionnaire was completed by 61 youth with T1D, and their PA was assessed. Hypoglycemia episodes, fear of hypoglycemia scores and HbA1C levels were collected. All assessments were realized at baseline (visit 0: V0), one year (V1) and two years later (V2). For the purpose of the present work, we explore the relationships between PA levels, hypoglycemia episodes, and FOH scores at each time point. We used multiple linear regression to model the mean outcomes for each exposure of interest. Results: Findings indicate no changes in total moderate to vigorous PA (MVPA) and VPA levels among visits, and HbA1c (%) was negatively correlated with the total amount of VPA per day in minutes (β= -0.44; p=0.01, β= -0.37; p=0.04, and β= -0.66; p=0.01 for V0, V1, and V2, respectively). Our linear regression model reported a significant negative correlation between VPA and FOH across the visits (β=-0.59, p=0.01; β= -0.44, p=0.01; and β= -0.34, p=0.03 for V0, V1, and V2, respectively), and HbA1c (%) was influenced by both the number of hypoglycemic episodes and FOH score at V2 (β=0.48; p=0.02 and β=0.38; p=0.03, respectively). Conclusion: The sustainability of PA levels and HbA1c (%) in young individuals with type 1 diabetes is influenced by various factors, including fear of hypoglycemia. Understanding these complex interactions is essential for developing effective interventions to promote sustained PA levels in this population. Our results underline the necessity of a multi-strategic approach to promoting active lifestyles among diabetic youths. This approach should synergize PA enhancement with vigilant glucose monitoring and effective FOH management.

Keywords: physical activity, hypoglycemia, fear of hypoglycemia, youth

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Authors: Xin Qi Dong, Melissa Simon

Abstract:

Aims: Elder mistreatment and suicidal ideation are important public health concerns among aging populations. This study will examine the association between elder mistreatment and suicidal ideation among Chinese older adults in the USA. Methods: Guided by a community-based participatory research approach, in this study we conducted in-person interviews with Chinese older adults aged 60 years and older in the Greater Chicago area from 2011 to 2013. Elder mistreatment was assessed by a 10-item instrument derived from the Hwalek-Sengstock Elder Abuse Screening Test (H-S/EAST) and the Vulnerability to Abuse Screening Scale (VASS). Suicidal ideation was assessed by the ninth item of the Patient Health Questionnaire-9 (PHQ-9) and the Geriatric Mental State Examination-Version A (GMS-A). Results: Overall, 3,159 Chinese older adults participated in this study, and their mean age was 72.8 years. After controlling for age, gender, education, income, medical comorbidities, depressive symptoms, and social support, elder mistreatment was significantly associated with 2-week suicidal ideation (OR 2.46, 95% CI 1.52--4.01) and 12-month suicidal ideation (OR 2.46, 95% CI 1.62--3.73). With respect to gender differences, the study found that the association remained significant for older women but not for older men after adjusting for all confounding factors. Conclusion: As the largest epidemiology study conducted among Chinese older adults in the USA, this study suggests that elder mistreatment is significantly associated with 2-week and 12-month suicidal ideation in older women but not in older men. Longitudinal studies should be conducted to explore the mechanisms through which elder mistreatment links with suicidal ideation.

Keywords: suicidal ideation, elder abuse, family violence, Asian health equity

Procedia PDF Downloads 227

Authors: Huajuan Shi

Abstract:

Background: The biopsy of the preimplantation embryo may increase the potential risk and concern of embryo viability. Clinically discarded spent embryo medium (SEM) has entered the view of researchers, sparking an interest in noninvasive embryo screening. However, one of the major restrictions is the extremelty low quantity of cf-RNA, which is difficult to efficiently and unbiased amplify cf-RNA using traditional methods. Hence, there is urgently need to an efficient and low bias amplification method which can comprehensively and accurately obtain cf-RNA information to truly reveal the state of SEM cf-RNA. Result: In this present study, we established an agarose PCR amplification system, and has significantly improved the amplification sensitivity and efficiency by ~90 fold and 9.29 %, respectively. We applied agarose to sequencing library preparation (named AG-seq) to quantify and characterize cf-RNA in SEM. The number of detected cf-RNAs (3533 vs 598) and coverage of 3' end were significantly increased, and the noise of low abundance gene detection was reduced. The increasing percentage 5' end adenine and alternative splicing (AS) events of short fragments (< 400 bp) were discovered by AG-seq. Further, the profiles and characterizations of cf-RNA in spent cleavage medium (SCM) and spent blastocyst medium (SBM) indicated that 4‐mer end motifs of cf-RNA fragments could remarkably differentiate different embryo development stages. Significance: This study established an efficient and low-cost SEM amplification and library preparation method. Not only that, we successfully described the characterizations of SEM cf-RNA of preimplantation embryo by using AG-seq, including abundance features fragment lengths. AG-seq facilitates the study of cf-RNA as a noninvasive embryo screening biomarker and opens up potential clinical utilities of trace samples.

Keywords: cell-free RNA, agarose, spent embryo medium, RNA sequencing, non-invasive detection

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Authors: Rajesh Kumar, Anil Kamboj

Abstract:

Inhibition of α- amylase play a vital role in the clinical management of postprandial hyperglycemia. Although, powerful synthetic inhibitors are available, natural inhibitors are potentially safer. The present study was carried out to evaluate α- amylase inhibition activity from hydroalcoholic extracts from aerial parts of Cestrum nocturnum. Hydroalcoholic extract was prepared by Soxhletation Method. The extract showed strong inhibition towards α- amylase activity and IC50 value were 45.9 µg. This In vitro studies indicate the potential of C. nocturnum in the development of effective anti-diabetic agents.

Keywords: α- amylase, cestrum nocturnum, hyperglycemia, hydroalcoholic extracts, diabetes

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Authors: Neelofar, Khursheed Alam, Jamal Ahmad

Abstract:

Protein modification in diabetes mellitus may lead to early glycation products (EGPs) or amadori product as well as advanced glycation end products (AGEs). Early glycation involves the reaction of glucose with N-terminal and lysyl side chain amino groups to form Schiff’s base which undergoes rearrangements to form more stable early glycation product known as Amadori product. After Amadori, the reactions become more complicated leading to the formation of advanced glycation end products (AGEs) that interact with various AGE receptors, thereby playing an important role in the long-term complications of diabetes. Millard reaction or nonenzymatic glycation reaction accelerate in diabetes due to hyperglycation and alter serum protein’s structure, their normal functions that lead micro and macro vascular complications in diabetic patients. In this study, Human Serum Albumin (HSA) with a constant concentration was incubated with different concentrations of glucose at 370C for a week. At 4th day, Amadori product was formed that was confirmed by colorimetric method NBT assay and TBA assay which both are authenticate early glycation product. Conformational changes in native as well as all samples of Amadori albumin with different concentrations of glucose were investigated by various biophysical and biochemical techniques. Main biophysical techniques hyperchromacity, quenching of fluorescence intensity, FTIR, CD and SDS-PAGE were used. Further conformational changes were observed by biochemical assays mainly HMF formation, fructoseamine, reduction of fructoseamine with NaBH4, carbonyl content estimation, lysine and arginine residues estimation, ANS binding property and thiol group estimation. This study find structural and biochemical changes in Amadori modified HSA with normal to hyperchronic range of glucose with respect to native HSA. When glucose concentration was increased from normal to chronic range biochemical and structural changes also increased. Highest alteration in secondary and tertiary structure and conformation in glycated HSA was observed at the hyperchronic concentration (75mM) of glucose. Although it has been found that Amadori modified proteins is also involved in secondary complications of diabetes as AGEs but very few studies have been done to analyze the conformational changes in Amadori modified proteins due to early glycation. Most of the studies were found on the structural changes in Amadori protein at a particular glucose concentration but no study was found to compare the biophysical and biochemical changes in HSA due to early glycation with a range of glucose concentration at a constant incubation time. So this study provide the information about the biochemical and biophysical changes occur in Amadori modified albumin at a range of glucose normal to chronic in diabetes. Although many implicates currently in use i.e. glycaemic control, insulin treatment and other chemical therapies that can control many aspects of diabetes. However, even with intensive use of current antidiabetic agents more than 50 % of diabetic patient’s type 2 suffers poor glycaemic control and 18 % develop serious complications within six years of diagnosis. Experimental evidence related to diabetes suggests that preventing the nonenzymatic glycation of relevant proteins or blocking their biological effects might beneficially influence the evolution of vascular complications in diabetic patients or quantization of amadori adduct of HSA by authentic antibodies against HSA-EGPs can be used as marker for early detection of the initiation/progression of secondary complications of diabetes. So this research work may be helpful for the same.

Keywords: diabetes mellitus, glycation, albumin, amadori, biophysical and biochemical techniques

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Authors: Julia R. Beulig, Stefan Ohla, Detlev Belder

Abstract:

In contrast to off-line analytical methods, lab-on-a-chip technology delivers direct information about the observed reaction. Therefore, microfluidic devices make an important scientific contribution, e.g. in the field of synthetic chemistry. Herein, the rapid generation of analytical data can be applied for the optimization of chemical reactions. These microfluidic devices enable a fast change of reaction conditions as well as a resource saving method of operation. In the presented work, we focus on the investigation of multiphase regimes, more specifically on a biphasic microfluidic droplet systems. Here, every single droplet is a reaction container with customized conditions. The biggest challenge is the rapid qualitative and quantitative readout of information as most detection techniques for droplet systems are non-specific, time-consuming or too slow. An exception is the electrospray mass spectrometry (ESI-MS). The combination of a reaction screening platform with a rapid and specific detection method is an important step in droplet-based microfluidics. In this work, we present a novel approach for synthesis optimization on the nanoliter scale with direct ESI-MS detection. The development of a droplet-based microfluidic device, which enables the modification of different parameters while simultaneously monitoring the effect on the reaction within a single run, is shown. By common soft- and photolithographic techniques a polydimethylsiloxane (PDMS) microfluidic chip with different functionalities is developed. As an interface for the MS detection, we use a steel capillary for ESI and improve the spray stability with a Teflon siphon tubing, which is inserted underneath the steel capillary. By optimizing the flow rates, it is possible to screen parameters of various reactions, this is exemplarity shown by a Domino Knoevenagel Hetero-Diels-Alder reaction. Different starting materials, catalyst concentrations and solvent compositions are investigated. Due to the high repetition rate of the droplet production, each set of reaction condition is examined hundreds of times. As a result, of the investigation, we receive possible reagents, the ideal water-methanol ratio of the solvent and the most effective catalyst concentration. The developed system can help to determine important information about the optimal parameters of a reaction within a short time. With this novel tool, we make an important step on the field of combining droplet-based microfluidics with organic reaction screening.

Keywords: droplet, mass spectrometry, microfluidics, organic reaction, screening

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Authors: Luiz Carlos Wrobel, Matjaz Hribersek, Jure Marn, Jurij Iljaz

Abstract:

Dynamic thermography has been clinically proven to be a valuable diagnostic technique for skin tumor detection as well as for other medical applications such as breast cancer diagnostics, diagnostics of vascular diseases, fever screening, dermatological and other applications. Thermography for medical screening can be done in two different ways, observing the temperature response under steady-state conditions (passive or static thermography), and by inducing thermal stresses by cooling or heating the observed tissue and measuring the thermal response during the recovery phase (active or dynamic thermography). The numerical modelling of heat transfer phenomena in biological tissue during dynamic thermography can aid the technique by improving process parameters or by estimating unknown tissue parameters based on measured data. This paper presents a nonlinear numerical model of multilayer skin tissue containing a skin tumor, together with the thermoregulation response of the tissue during the cooling-rewarming processes of dynamic thermography. The model is based on the Pennes bioheat equation and solved numerically by using a subdomain boundary element method which treats the problem as axisymmetric. The paper includes computational tests and numerical results for Clark II and Clark IV tumors, comparing the models using constant and temperature-dependent thermophysical properties, which showed noticeable differences and highlighted the importance of using a local thermoregulation model.

Keywords: boundary element method, dynamic thermography, static thermography, skin tumor diagnostic

Procedia PDF Downloads 107

Authors: Huajuan Shi

Abstract:

Background: The biopsy of the preimplantation embryo may increase the potential risk and concern of embryo viability. Clinically discarded spent embryo medium (SEM) has entered the view of researchers, sparking an interest in noninvasive embryo screening. However, one of the major restrictions is the extremelty low quantity of cf-RNA, which is difficult to efficiently and unbiased amplify cf-RNA using traditional methods. Hence, there is urgently need to an efficient and low bias amplification method which can comprehensively and accurately obtain cf-RNA information to truly reveal the state of SEM cf-RNA. Result: In this present study, we established an agarose PCR amplification system, and has significantly improved the amplification sensitivity and efficiency by ~90 fold and 9.29 %, respectively. We applied agarose to sequencing library preparation (named AG-seq) to quantify and characterize cf-RNA in SEM. The number of detected cf-RNAs (3533 vs 598) and coverage of 3' end were significantly increased, and the noise of low abundance gene detection was reduced. The increasing percentage 5' end adenine and alternative splicing (AS) events of short fragments (< 400 bp) were discovered by AG-seq. Further, the profiles and characterizations of cf-RNA in spent cleavage medium (SCM) and spent blastocyst medium (SBM) indicated that 4‐mer end motifs of cf-RNA fragments could remarkably differentiate different embryo development stages. Significance: This study established an efficient and low-cost SEM amplification and library preparation method. Not only that, we successfully described the characterizations of SEM cf-RNA of preimplantation embryo by using AG-seq, including abundance features fragment lengths. AG-seq facilitates the study of cf-RNA as a noninvasive embryo screening biomarker and opens up potential clinical utilities of trace samples.

Keywords: cell-free RNA, agarose, spent embryo medium, RNA sequencing, non-invasive detection

Procedia PDF Downloads 64

Authors: Md. Fazlul Karim, Ashik Sharfaraz, Aysha Ferdoushi

Abstract:

Background: Computational medicinal chemistry approaches are used for designing and identifying new drug-like molecules, predicting properties and pharmacological activities, and optimizing lead compounds in drug development. SIRT7, a nicotinamide adenine dinucleotide (NAD+)-dependent deacylase which regulates aging, is an emerging target for cancer therapy with mounting evidence that SIRT7 downregulation plays important roles in reversing cancer phenotypes and suppressing tumor growth. Activation or altered expression of SIRT7 is associated with the progression and invasion of various cancers, including liver, breast, gastric, prostate, and non-small cell lung cancer. Objectives: The goal of this work was to identify potent and selective bioactive candidate inhibitors of SIRT7 by in silico screening of small molecule compounds obtained from Nigella sativa (N. sativa). Methods: SIRT7 structure was retrieved from The Research Collaboratory for Structural Bioinformatics Protein Data Bank (RCSB PDB), and its active site was identified using CASTp and metaPocket. Molecular docking simulation was performed with PyRx 0.8 virtual screening software. Drug-likeness properties were tested using SwissADME and pkCSM. In silico toxicity was evaluated by Osiris Property Explorer. Bioactivity was predicted by Molinspiration software. Antitumor activity was screened for Prediction of Activity Spectra for Substances (PASS) using Way2Drug web server. Molecular dynamics (MD) simulation was carried out by Desmond v3.6 package. Results: A total of 159 bioactive compounds from the N. Sativa were screened against the SIRT7 enzyme. Five bioactive compounds: chrysin (CID:5281607), pinocembrin (CID:68071), nigellidine (CID:136828302), nigellicine (CID:11402337), and epicatechin (CID:72276) were identified as potent SIRT7 anti-cancer candidates after docking score evaluation and applying Lipinski's Rule of Five. Finally, MD simulation identified Chrysin as the top SIRT7 anti-cancer candidate molecule. Conclusion: Chrysin, which shows a potential inhibitory effect against SIRT7, can act as a possible anti-cancer drug candidate. This inhibitor warrants further evaluation to check its pharmacokinetics and pharmacodynamics properties both in vitro and in vivo.

Keywords: SIRT7, antitumor, molecular docking, molecular dynamics simulation

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