Search results for: protein malnutrition

Authors: Wirot Likittrakulwong, Pisit Poolprasert, Tossaporn Incharoen

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Germinated paddy rice (GPR) has the potential to be used as a feed ingredient. However, their properties have not been fully investigated. This paper examined the nutrient digestibility and the relationship to immune activity in Roman hens fed with GPR. It was found that true and apparent metabolizable energy (ME) values of GPR were 3.20 and 3.28 kcal/g air dry, respectively. GPR exhibited high content of phytonutrients, especially GABA. GPR showed similar protein profiles in comparison to non-germinated paddy rice. For immune activity, the feed with GPR enhanced the immune activity of Roman hens under high stocking density stress as evidenced by the activity of superoxide dismutase (SOD) and lysozyme activity. In this study, GPR is proved to be a good source of functional ingredient for chicken feed.

Keywords: germinated paddy rice, nutrient digestibility, immune activity, functional property

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Authors: Oktia Woro Kasmini Handayani, Bambang Budi Raharjo, Efa Nugroho, Bertakalswa Hermawati

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Obesity prevalence and severe malnutrition in Indonesia has increased from 2007 to 2013. The utilization of local resources in nutritional program planning can be used to program efficiency and to reach the goal. The aim of this research is to plan a nutrition program based on local resources for urban fringe areas in a developing country. This research used a qualitative approach, with a focus on local resources including social capital, social system, cultural system. The study was conducted in Mijen, Central Java, as one of the urban fringe areas in Indonesia. Purposive and snowball sampling techniques are used to determine participants. A total of 16 participants took part in the study. Observation, interviews, focus group discussion, SWOT analysis, brainstorming and Miles and Huberman models were used to analyze the data. We have identified several local resources, such as the contributions from nutrition cadres, social organizations, social financial resources, as well as the cultural system and social system. The outstanding contribution of nutrition cadres is the participation and creativity to improve nutritional status. In addition, social organizations, like the role of the integrated health center for children (Pos Pelayanan Terpadu), can be engaged in the nutrition program planning. This center is supported by House of Nutrition to assist in nutrition program planning, and provide social support to families, neighbors and communities as social capitals. The study also reported that cultural systems that show appreciation for well-nourished children are a better way to improve the problem of balanced nutrition. Social systems such as teamwork and mutual cooperation can also be a potential resource to support nutritional programs and overcome associated problems. The impact of development in urban areas such as the introduction of more green areas which improve the perceived status of local people, as well as new health services facilitated by people and companies, can also be resources to support nutrition programs. Local resources in urban fringe areas can be used in the planning of nutrition programs. The expansion of partnership with all stakeholders, empowering the community through optimizing the roles of nutrition care centers for children as our recommendation with regard to nutrition program planning.

Keywords: developing country, local resources, nutrition program, urban fringe

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Authors: Vanessa Funk, Steven Blum, Stephanie Cole, Jorge Maciel, Matthew Lux

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Paper-based synthetic gene circuits offer a new paradigm for programmable, fieldable biodetection. We demonstrate that by freeze-drying gene circuits with in vitro expression machinery, we can use complimentary RNA sequences to trigger colorimetric changes upon rehydration. We have successfully utilized both green fluorescent protein and luciferase-based reporters for easy visualization purposes in solution. Through several efforts, we are aiming to use this new platform technology to address a variety of needs in portable detection by demonstrating several more expression and reporter systems for detection functions on paper. In addition to RNA-based biodetection, we are exploring the use of various mechanisms that cells use to respond to environmental conditions to move towards all-hazards detection. Examples include explosives, heavy metals for water quality, and toxic chemicals.

Keywords: cell-free lysates, detection, gene circuits, in vitro

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Authors: Mesfin Angaw Tesfay

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Uridine-5’-diphosphate (UDP)-glucose is one of the most versatile building blocks within the metabolism of prokaryotes and eukaryotes serving as an activated sugar donor during the glycosylation of natural products. It is formed by the enzyme UDP-glucose pyrophosphorylase (UGPase) using uridine-5′-triphosphate (UTP) and α-d-glucose 1-phosphate as a substrate. Herein two UGPase genes from Lactobacillus kefiranofaciens ZW3 (LkUGPase) and Rhodococcus wratislaviensis IFP 2016 (RwUGPase) were identified through genome mining approaches. The LkUGPase and RwUGPase have 299 and 306 amino acids, respectively. Both UGPase has the conserved UTP binding site (G-X-G-T-R-X-L-P) and the glucose -1-phosphate binding site (V-E-K-P). The LkUGPase and RwUGPase were cloned in E. coli and SDS-PAGE analysis showed the expression of both enzymes forming about 36 KDa of protein band after induction. LkUGPase and RwUGPase have an activity of 1549.95 and 671.53 U/mg respectively. Currently, their kinetic properties are under investigation.

Keywords: UGPase, LkUGPase, RwUGPase, UDP-glucose, Glycosylation

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Authors: M. Akoz, O. B. Citil, I. Aydin

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Colostrum secreted by the mammary glands after birth in the early days, a high proportion of fat, protein and ash containing a secretion containing low amounts of casein and lactose. Especially immunoglobulins contain high proportions. Maternal immunoglobulins own immune system to protect the newborn against neonatal disease until development are very important matter. However, colostrum is transferred to the offspring due to placental barrier in ruminants. Immunoglobulins are absorbed through the intestinal epithelium but absorption can vary under the influence of some factors. These factors are among the priority ones taking colostrum first time, amount, concentration, the metabolic status of the newborn. intestinal absorption of immunoglobulins occurs over the first 24 h high. Absorption from the gut after nine hours, 50% after 24 hours was only 11%. On the other hand pup's digestive system degrade the enzymes after 24 hours immunoglobulins. Bovine colostrum in the composition while basic immune IgG, IgA and IgM are also available. Total IgG in colostrum of ruminants, while in other species is a greater amount in blood serum.

Keywords: immunoglobulin, ruminants, colostrum, immune system

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Authors: Walter Vera-Ortega, Idoia Busnadiego, Sam J. Wilson

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Introduction: Human Immunodeficiency Virus type 1 (HIV-1) is responsible for the acquired immunodeficiency syndrome (AIDS), a leading cause of death worldwide infecting millions of people each year. Despite intensive research in vaccine development, therapies against HIV-1 infection are not curative, and the huge genetic variability of HIV-1 challenges to drug development. Current animal models for HIV-1 research present important limitations, impairing the progress of in vivo approaches. Macaques require a CD8+ depletion to progress to AIDS, and the maintenance cost is high. Mice are a cheaper alternative but need to be 'humanized,' and breeding is not possible. The development of an HIV-1 clone able to replicate in mice is a challenging proposal. The lack of human co-factors in mice impedes the function of the HIV-1 accessory proteins, Tat and Rev, hampering HIV-1 replication. However, Tat and Rev function can be replaced by constitutive/chimeric promoters, codon-optimized proteins and the constitutive transport element (CTE), generating a novel HIV-1 clone able to replicate in mice without disrupting the amino acid sequence of the virus. By minimally manipulating the genomic 'identity' of the virus, we propose the generation of an HIV-1 clone able to replicate in mice to assist in antiviral drug development. Methods: i) Plasmid construction: The chimeric promoters and CTE copies were cloned by PCR using lentiviral vectors as templates (pCGSW and pSIV-MPCG). Tat mutants were generated from replication competent HIV-1 plasmids (NHG and NL4-3). ii) Infectivity assays: Retroviral vectors were generated by transfection of human 293T cells and murine NIH 3T3 cells. Virus titre was determined by flow cytometry measuring GFP expression. Human B-cells (AA-2) and Hela cells (TZMbl) were used for infectivity assays. iii) Protein analysis: Tat protein expression was determined by TZMbl assay and HIV-1 capsid by western blot. Results: We have determined that NIH 3T3 cells are able to generate HIV-1 particles. However, they are not infectious, and further analysis needs to be performed. Codon-optimized HIV-1 constructs are efficiently made in 293T cells in a Tat and Rev independent manner and capable of packaging a competent genome in trans. CSGW is capable of generating infectious particles in the absence of Tat and Rev in human cells when 4 copies of the CTE are placed preceding the 3’LTR. HIV-1 Tat mutant clones encoding different promoters are functional during the first cycle of replication when Tat is added in trans. Conclusion: Our findings suggest that the development of an HIV-1 Tat-Rev independent clone is challenging but achievable aim. However, further investigations need to be developed prior presenting our HIV-1 clone as a candidate model for research.

Keywords: codon-optimized, constitutive transport element, HIV-1, long terminal repeats, research model

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Authors: Nouha Bouayed Abdelmoula, Rim Louati, Nawel Abdellaoui, Balkiss Abdelmoula, Oldez Kaabi, Walid Smaoui, Samir Aloulou

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Background and Aims: Cardiofaciocutaneous syndrome (CFC) is an autosomal dominant disorder with the vast majority of cases arising by a new mutation of BRAF, MEK1, MEK2, or rarely, KRAS genes. Here, we report a rare Tunisian case of CFC syndrome for whom we identify SOS1 mutation. Methods: Genomic DNA was obtained from peripheral blood collected in an EDTA tube and extracted from leukocytes using the phenol/chloroform method according to standard protocols. High resolution melting (HRM) analysis for screening of mutations in the entire coding sequence of PTPN11 was conducted first. Then, HRM assays to look for hot spot mutations coding regions of the other genes of the RAS-MAPK pathway (RAt Sarcoma viral oncogene homolog Mitogen-Activated Protein Kinases Pathway): SOS1, SHOC2, KRAS, RAF1, KRAS, NRAS, CBL, BRAF, MEK1, MEK2, HRAS, and RIT1, were applied. Results: Heterozygous SOS1 point mutation clustered in exon 10, which encodes for the PH domain of SOS1, was identified: c.1655 G > A. The patient was a 9-year-old female born from a consanguineous couple. She exhibited pulmonic valvular stenosis as congenital heart disease. She had facial features and other malformations of Noonan syndrome, including macrocephaly, hypertelorism, ptosis, downslanting palpebral fissures, sparse eyebrows, a short and broad nose with upturned tip, low-set ears, high forehead commonly associated with bitemporal narrowing and prominent supraorbital ridges, short and/or webbed neck and short stature. However, the phenotype is also suggestive of CFC syndrome with the presence of more severe ectodermal abnormalities, including curly hair, keloid scars, hyperkeratotic skin, deep plantar creases, and delayed permanent dentition with agenesis of the right maxillary first molar. Moreover, the familial history of the patient revealed recurrent brain malignancies in the paternal family and epileptic disease in the maternal family. Conclusions: This case report of an overlapping RASopathy associated with SOS1 mutation and familial history of brain tumorigenesis is exceptional. The evidence suggests that RASopathies are truly cancer-prone syndromes, but the magnitude of the cancer risk and the types of cancer partially overlap.

Keywords: cardiofaciocutaneous syndrome, CFC, SOS1, brain cancer, germline mutation

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Authors: Hategekimana Jean Paul, Irakoze Josiane, Ishimweyizerwe Valentin, Iradukunda Dieudonne, Uwanyirigira Jeannette

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Orange fleshed sweet potatoes (OFSP) are highly grown and are available plenty in rural and urban local markets and its contribution in reduction of food insecurity in Rwanda is considerable. But the postharvest loss of this commodity is a critical challenge due to its high perishability. Several research activities have been conducted on how fresh food commodities can be transformed into extended shelf life food products for prevention of post-harvest losses. However, such activity was not yet well studied in Rwanda. The aim of the present study was the processing of backed products from (OFSP)combined with wheat composite flour and assess the nutritional content and consumer acceptability of new developed products. The perishability of OFSP and their related lack during off season can be eradicated by producing cake, doughnut and bread with OFSP puree or flour. The processing for doughnut and bread were made by making OFSP puree and other ingredients then a dough was made followed by frying and baking while for cake OFSP was dried through solar dryer to have a flour together with wheat flour and other ingredients to make dough cake and baking. For each product, one control and three experimental samples, (three products in three different ratios (30,40 and50%) of OFSP and the remaining percentage of wheat flour) were prepared. All samples including the control were analyzed for the consumer acceptability (sensory attributes). Most preferred samples (One sample for each product with its control sample and for each OFSP variety) were analyzed for nutritional composition along with control sample. The Cake from Terimbere variety and Bread from Gihingumukungu supplemented with 50% OFSP flour or Puree respectively were most acceptable except Doughnut from Vita variety which was highly accepted at 50% of OFSP supplementation. The moisture, ash, protein, fat, fiber, Total carbohydrate, Vitamin C, reducing sugar and minerals (Sodium, Potassium and Phosphorus.) content was different among products. Cake was rich in fibers (14.71%), protein (6.590%), and vitamin c(19.988mg/100g) compared to other samples while bread found to be rich in reducing sugar with 12.71mg/100g compared to cake and doughnut. Also doughnut was found to be rich in fat content with 6.89% compared to other samples. For sensory analysis, doughnut was highly accepted in ratio of 60:40 compared to other products while cake was least accepted at ratio of 50:50. The Proximate composition and minerals content of all the OFSP products were significantly higher as compared to the control samples.

Keywords: post-harvest loss, OFSP products, wheat flour, sensory evaluation, proximate composition

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Authors: V. Villa-Cruz, H. Pérez-Ladron de Guevara, J. E. Diaz-Díaz

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Static Light Scattering (SLS) was used as a method to analyse cow's milk raw, coming from the town of Lagos de Moreno, Jalisco, Mexico. This method is based on the analysis of the dispersion of light laser produced by a set of particles in solution. Based on the above, raw milk, which contains particles of fat globules, with a diameter of 2000 nm and particles of micelles of protein with 300 nm in diameter were analyzed. For this, dilutions of commercial milk were made (1.0%, 2.0% and 3.3%) to obtain a pattern of laser light scattering and also made measurements of raw cow's milk. Readings were taken in a sweep initial angle 10° to 170°, results were analyzed with the program OriginPro 7. The SLS method gives us an estimate of the percentage of fat content in milk samples. It can be concluded that the SLS method, is a quick method of analysis to detect adulteration in raw cow's milk.

Keywords: light scattering, milk analysis, adulteration in milk, micelles, OriginPro

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Authors: Asim Cengiz, Dede Basturk, Hakan Ozalp

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Practicing recurring resistance workout such as may cause changes in human muscle. These changes may be because combination if several factors determining physical fitness. Thus, it is important to identify these changes. Several studies were reviewed to investigate these changes. As a result, the changes included positive modifications in amplified citrate synthase (CS) maximal activity, increased capacity for pyruvate oxidation, improvement on molecular signaling on human performance, amplified resting muscle glycogen and whole GLUT4 protein content, better health outcomes such as enhancement in cardiorespiratory fitness. Sprint training also have numerous long long-term changes inhuman body such as better enzyme action, changes in muscle fiber and oxidative ability. This is important because SV is the critical factor influencing maximal cardiac output and therefore oxygen delivery and maximal aerobic power.

Keywords: sprint, training, performance, exercise

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Authors: Mohamed Fawzy Elzarei, Yousef Mohammed Al-Dakheel, Ali Mohamed Alseaf

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Modern molecular techniques, like single marker analysis for linked traits to these markers, can provide us with rapid and accurate genetic results. In the last two decades of the last century, the applications of molecular techniques were reached a faraway point in cattle, sheep, and pig. In goats, especially in our region, the application of molecular techniques is still far from other species. As reported by many researchers, microsatellites marker is one of the suitable markers for lie studies. The single marker linked to traits of interest is one technique allowed us to early select animals without the necessity for mapping the entire genome. Simplicity, applicability, and low cost of this technique gave this technique a wide range of applications in many areas of genetics and molecular biology. Also, this technique provides a useful approach for evaluating genetic differentiation, particularly in populations that are poorly known genetically. The expected breeding value (EBV) and yield deviation (YD) are considered as the most parameters used for studying the linkage between quantitative characteristics and molecular markers, since these values are raw data corrected for the non-genetic factors. A total of 17 microsatellites markers (from chromosomes 6, 14, 18, 20 and 23) were used in this study to search for areas that could be responsible for genetic variability for some milk traits and search of chromosomal regions that explain part of the phenotypic variance. Results of single-marker analyses were used to identify the linkage between microsatellite markers and variation in EBVs of these traits, Milk yield, Protein percentage, Fat percentage, Litter size and weight at birth, and litter size and weight at weaning. The estimates of the parameters from forward and backward solutions using stepwise regression procedure on milk yield trait, only two markers, OARCP9 and AGLA29, showed a highly significant effect (p≤0.01) in backward and forward solutions. The forward solution for different equations conducted that R2 of these equations were highly depending on only two partials regressions coefficient (βi,) for these markers. For the milk protein trait, four marker showed significant effect BMS2361, CSSM66 (p≤0.01), BMS2626, and OARCP9 (p≤0.05). By the other way, four markers (MCM147, BM1225, INRA006, andINRA133) showed highly significant effect (p≤0.01) in both backward and forward solutions in association with milk fat trait. For both litter size at birth and at weaning traits, only one marker (BM143(p≤0.01) and RJH1 (p≤0.05), respectively) showed a significant effect in backward and forward solutions. The estimates of the parameters from forward and backward solution using stepwise regression procedure on litter weight at birth (LWB) trait only one marker (MCM147) showed highly significant effect (p≤0.01) and two marker (ILSTS011, CSSM66) showed a significant effect (p≤0.05) in backward and forward solutions.

Keywords: microsatellites marker, estimated breeding value, stepwise regression, milk traits

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Authors: Tiziana Pepe, Gerardo Manfreda, Adriana Ianieri, Aniello Anastasio

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The sustainable exploitation of marine biological resources is among the most important objectives of the EU's Common Fisheries Policy (CFP). Currently, Europe imports about 65% of its fish products, indicating that domestic production does not meet consumer demand. Despite the availability of numerous commercially significant fish species, European consumption is concentrated on a limited number of products (e.g., sea bass, sea bream, shrimp). Many native species, present in large quantities in the Mediterranean Sea, are little known to consumers and are therefore considered ‘fishing by-products’. All the data presented so far indicate a significant waste of local resources and the overexploitation of a few fish stocks. It is therefore necessary to develop strategies that guide the market towards sustainable conversion. The objective of this work was to valorize underutilized fish species of the Mediterranean Sea through a multidisciplinary approach. To this end, three fish species were sampled: Atlantic Horse Mackerel (Trachurus trachurus), Bogue (Boops boops), and Common Dolphinfish (Coryphaena hippurus). Nutritional properties (water %, fats, proteins, ashes, salts), physical/chemical properties (TVB-N, histamine, pH), and rheological properties (color, texture, viscosity) were analyzed. The analyses were conducted on both fillets and processing by-products. Additionally, mitochondrial DNA (mtDNA) was extracted from the muscle of each species. The mtDNA was then sequenced using the Illumina NGS technique. The analysis of nutritional properties classified the fillets of the sampled species as lean or semi-fat, as they had a fat content of less than 3%, while the by-products showed a higher lipid content (2.7-5%). The protein percentage for all fillets was 22-23%, while for processing by-products, the protein concentration was 18-19% for all species. Rheological analyses showed an increase in viscosity in saline solution in all species, indicating their potential suitability for industrial processing. High-quality and quantity complete mtDNA was extracted from all analyzed species. The complete mitochondrial genome sequences were successfully obtained and annotated. The results of this study suggest that all analyzed species are suitable for both human consumption and feed production. The sequencing of the complete mtDNA and its availability in international databases will be useful for accurate phylogenetic analysis and proper species identification, even in prepared and processed products. Underutilized fish species represent an important economic resource. Encouraging their consumption could limit the phenomenon of overfishing, protecting marine biodiversity. Furthermore, the valorization of these species will increase national fish production, supporting the local economy, cultural, and gastronomic tradition, and optimizing the exploitation of Mediterranean resources in accordance with the CFP.

Keywords: mtDNA, nutritional analysis, sustainable fisheries, underutilized fish species

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Authors: Simon Grossemy, Peggy P. Y. Chan, Pauline M. Doran

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Nerve tissue engineering is the main field of research aimed at finding an alternative to autografts as a treatment for nerve injuries. Scaffolds are used as a support to enhance nerve regeneration. In order to successfully design novel scaffolds and in vitro cell culture systems, a deep understanding of the factors affecting nerve regeneration processes is needed. Physical and biological parameters associated with the culture environment have been identified as potentially influential in nerve cell differentiation, including electrical stimulation, exposure to extracellular-matrix (ECM) proteins, dynamic medium conditions and co-culture with glial cells. The mechanisms involved in driving the cell to differentiation in the presence of these factors are poorly understood; the complexity of each of them raises the possibility that they may strongly influence each other. Some questions that arise in investigating nerve regeneration include: What are the best protein coatings to promote neural cell attachment? Is the scaffold design suitable for providing all the required factors combined? What is the influence of dynamic stimulation on cell viability and differentiation? In order to study these effects, scaffolds adaptable to bioreactor culture conditions were designed to allow electrical stimulation of cells exposed to ECM proteins, all within a dynamic medium environment. Gold coatings were used to make the surface of viscose rayon microfiber scaffolds (VRMS) conductive, and poly-L-lysine (PLL) and laminin (LN) surface coatings were used to mimic the ECM environment and allow the attachment of rat PC12 neural cells. The robustness of the coatings was analyzed by surface resistivity measurements, scanning electron microscope (SEM) observation and immunocytochemistry. Cell attachment to protein coatings of PLL, LN and PLL+LN was studied using DNA quantification with Hoechst. The double coating of PLL+LN was selected based on high levels of PC12 cell attachment and the reported advantages of laminin for neural differentiation. The underlying gold coatings were shown to be biocompatible using cell proliferation and live/dead staining assays. Coatings exhibiting stable properties over time under dynamic fluid conditions were developed; indeed, cell attachment and the conductive power of the scaffolds were maintained over 2 weeks of bioreactor operation. These scaffolds are promising research tools for understanding complex neural cell behavior. They have been used to investigate major factors in the physical culture environment that affect nerve cell viability and differentiation, including electrical stimulation, bioreactor hydrodynamic conditions, and combinations of these parameters. The cell and tissue differentiation response was evaluated using DNA quantification, immunocytochemistry, RT-qPCR and functional analyses.

Keywords: bioreactor, electrical stimulation, nerve differentiation, PC12 cells, scaffold

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Authors: Syed Sameer Aga, Saniya Nissar

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The Xeroderma pigmentosum group D gene (XPD) plays a key role in nucleotide excision repair (NER) pathway of the damaged DNA. Genetic polymorphisms in the coding region of the XPD gene may alter DNA repair capacity of the protein and hence can modulate the risk of colorectal cancer (CRC) risk. The aim of the study was to determine the genetic association of XPD Lys751Gln polymorphism with the risk of colorectal cancer (CRC) development. 120 CRC patients and 160 normal controls were assessed for genotype frequencies of XPD Lys751Gln polymorphism using PCR-RFLP technique. We observed a significant association (p < 0.05) between the XPD Lys751Gln polymorphism and the risk of developing CRC (p < 0.05). Additionally, Gln/Gln genotype of the XPD gene doubled the risk for the development of CRC [p < 0.05; OR=2.25 95% CI (1.07-4.7)]. Our results suggest that there is a significant association between the XPD Lys751Gln polymorphism and the risk of CRC.

Keywords: colorectal cancer, polymorphism, RFLP, DNA Repair, NER, XPD

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Authors: P. Vosynek, T. Návrat, M. Peč, J. Pořízka, P. Diviš

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Changes of mechanical properties and response behavior of bones is an important external sign of medical problems like osteoporosis, bone remodeling after fracture or surgery, osteointegration, or bone tissue loss of astronauts in space. Measuring of mechanical behavior of bones in physiological and osteoporotic states, quantified by different degrees of protein (collagen) and mineral loss, is thus an important topic in biomechanical research. This contribution deals with the relation between mechanical properties of the turkey long bone–tibia in physiological, demineralized, and deproteinized state. Three methods for comparison were used: densitometry, three point bending and harmonic response analysis. The results help to find correlations between the methods and estimate their possible application in medical practice.

Keywords: bone properties, long bone, osteoporosis, response behavior

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Authors: Tiancheng Lan

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E10A is a kind of replication-defective adenovirus which carries the human endostatin gene to inhibit the growth of tumors. Kringle 5(K5) has almost the same function as angiostatin to also inhibit the growth of tumors since they are all the byproduct of the proteolytic cleavage of plasminogen. Tumor size increasing can be suppressed because both of the endostatin and K5 can restrain the angiogenesis process. Therefore, in order to improve the treatment effect on tumor, 2A peptide is used to construct a fusion gene carrying both E10A and K5. Using 2A peptide is an ideal strategy when a fusion gene is expressed because it can avoid many problems during the expression of more than one kind of protein. The overlap extension PCR is also used to connect 2A peptide with E10A and K5. The final construction of fusion gene E10A-2A-K5 can provide a possible new method of the anti-angiogenesis treatment with a better expression performance.

Keywords: E10A, Kringle 5, 2A peptide, overlap extension PCR

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Authors: Jin Li, Jing Sun, Xiangkun Cai, Lijuanwang, Yanbin Tang, Junsheng Huo

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In order to improve the malnutrition of infants and young children in poor rural areas of China, Chinese government implement a project on improvement of children's nutrition in poor rural areas. Each infant or young child aged 6 to 23 months in selected poor rural areas of China was provided a package of Yingyangbao (YYB) per day, which is a full fat soy powder mixed with multiple micronutrient powders. A technical direction to implement this project comprehensively in poor rural areas of China will be provided by assessing the nutritional status of infants and feeding practices of caregiver. The nutritional intervention was conducted using Yingyangbao for infants aged 6 to 23 months in six poor counties of Shanxi, Yunnan and Hubei Provinces. The caregiver or parents of infants were educated on feeding knowledge and practice. A total of 1840 infants were assessed before the intervention and 1789 infants one year later. The length, weight, hemoglobin concentration of infants were measured to evaluate nutritional status before and after the intervention respectively. The questionnaires were designed to collect data for the basic demographic information and feeding practices. The average weight of infants aged 6 to 23 months increased from 9.59 ± 1.54kg to 9.73 ± 1.61kg one years later (p<0.01), and the average length from 76.0±6.0 to 77.0±6.1(p<0.01). The weight and length of infants aged 12 to 17 months had most obviously improving effect among the three age groups. Before the intervention, the hemoglobin concentration value of infants was 11.7±1.2g/L, and the anemia prevalence was 32.9%. One year later, the hemoglobin concentration value of the infants was increased to 12.0±1.1g/dL, and the anemia prevalence was decreased to 26.0%. There were both statistically significant (p <0.01). The anemia prevalence of infants aged 18 to 23 months had most obviously improving effect，which decreased from 25.0% to 17.2%(p<0.01). The proportion of infants aged 6 to 8 months who received solid, semi-solid or soft foods in time was increased from 89.4% to 91.6%, while there was no statistically significant. The proportion of 6-23 month-old infants who received minimum dietary diversity increased from 55.6% to 60.3%(p <0.01). The differences of the proportion of infants who received minimum meal frequency was no statistically significant between before and after the intervention. The nutritional intervention using Yingyangbao showed the significant effect for improving infants aged 6 to 23 months anemia status, weight and length. The feeding practices were improved through education in the process of nutritional intervention, while the effect is not significant. It is need for Chinese government to explore new publicity pattern.

Keywords: nutritional intervention, infants, nutritional status, feeding practice

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Authors: Petnamnueng Dettipponpong, Shuen E. Chen

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Heat stress is known to have negative effects on reproductive functions, such as follicular development and ovulation. This study aimed to investigate the specific effects of heat stress on steroid hormone secretion of ovarian follicle cells, particularly in relation to the expression of Apolipoprotein B (ApoB) and microsomal triglyceride transfer protein (MTP). The aim of the study was to understand the impact of heat stress on steroid hormone secretion in ovarian follicle cells and to explore the role of ApoB and MTP in this process. Primary granulosa and theca cells were collected from follicles and cultured under heat stress conditions (42 °C) for various time periods. Controls were maintained under normal conditions (37.5 °C ). The culture medium was collected at different time points to measure levels of progesterone and estradiol using ELISA kits. ApoB and MTP expression levels were analyzed using homemade antibodies and western blot. Data were assessed by a one-way ANOVA comparison test with Duncan’s new multiple-range test. Results were expressed as mean±S.E. Difference was considered significant at P<0.05. The results showed that heat stress significantly increased progesterone secretion in granulosa cells, with the peak observed after 13 hours of recovery under thermoneutral conditions. Estradiol secretion by theca cells was not affected. Heat stress also had a significant negative effect on granulosa cell viability. Additionally, the expression of ApoB and MTP was found to be differentially regulated by heat stress. ApoB expression in theca cells was transiently promoted, while ApoB expression in granulosa cells was consistently suppressed. MTP expression increased after 5 hours of recovery in both cell types. These findings suggest a mechanism by which chicken follicle cells export cellular lipids as very low-density lipoprotein (VLDL) in response to thermal stress. These contribute to our understanding of the role of ApoB and MTP steroidogenesis and lipid metabolism under heat stress conditions. The study involved the collection of primary granulosa and theca cells, culture under different temperature conditions, and analysis of the culture medium for hormone levels using ELISA kits. ApoB and MTP expression levels were assessed using homemade antibodies and western blot. This study aimed to address the effects of heat stress on steroid hormone secretion in ovarian follicle cells, as well as the role of ApoB and MTP in this process. The study demonstrates that heat stress stimulates steroidogenesis in granulosa cells, affecting progesterone secretion. ApoB and MTP expression were found to be differentially regulated by heat stress, indicating a potential mechanism for the export of cellular lipids in response to thermal stress.

Keywords: heat stress, granulosa cells, theca cells, steroidogenesis, chicken, apolipoprotein B, microsomal triglyceride transfer protein

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Authors: Bongmun Kang, Kagnari Yamakawa, Yoshihisa Hagihara, Yuji Ito, Michimasa Kishimoto, Yoichi Kumada

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The PMMA-tag was genetically fused with the C-terminal region of VHH molecules. This antibody, VHH, is known as a single-chain domain, which is devoid of light chains. The PMMA-tag, which could affect the isoelectric point (pI) changeable with a charge of amino acid in VHHs were closely related to the solubility of VHH molecules during refolding. The genetic fusion of PMMA-tag to C-terminal region of VHHs significantly affects the recovery of their soluble protein during refolding by 50 mM TAPS at pH 8.5. It could be refolded with a recovery of more than 95% by dialysis at pH 8.5. A marked difference in the antigen-binding activities in the adsorption state was significantly high in VHH-PM compared to the wild type of VHH. There are approximately 8-fold differences in the antigen-binding activities in the adsorption state between VHH-PM and VHH.

Keywords: VHH, PMMA-tag, isoelectric point, pH, Solubility, refolding, immobilization, ELISA

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Authors: Sudhir Kumar Sharma, Abiy D. Woldetsadik, Mazin Magzoub, Ramesh Jagannathan

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It is well known that bioactive ceramics exhibit specific biological affinities, especially in the area of tissue re-generation. In this context, we report the development of an eminently scalable, novel, supercritical CO₂ based process for the fabrication of hierarchically porous 'soft' CaCO₃ scaffolds. Porosity at the macro, micro, and nanoscales was obtained through process optimization of the so-called 'coffee-ring effect'. Exposure of these CaCO₃ scaffolds to monocytic THP-1 cells yielded negligible levels of tumor necrosis factor-alpha (TNF-α) thereby confirming the lack of immunogenicity of the scaffolds. ECM protein treatment of the scaffolds showed enhanced adsorption comparable to standard control such as glass. In vitro studies using osteoblast precursor cell line, MC3T3, also demonstrated the cytocompatibility of hierarchical porous CaCO₃ scaffolds.

Keywords: supercritical CO2, CaCO3 scaffolds, coffee-ring effect, ECM proteins

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Authors: Vasile Simulescu, Jakub Mondek, Miloslav Pekař

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Hyaluronic acid (HA) is an anionic glycosaminoglycan distributed throughout connective, epithelial and neural tissues. The importance of hyaluronic acid increased in the last decades. It has many applications in medicine and cosmetics. Hyaluronic acid has been used in attempts to treat osteoarthritis of the knee via injecting it into the joint. Bovine serum albumin (also known as BSA) is a protein derived from cows, which has many biochemical applications. The aim of our research work was to compare the thermal degradation of hyaluronic acid and BSA in powder and in solution, by determining changes in molar mass and conformation, by using SEC-MALLS (size exclusion chromatography -multi angle laser light scattering). The aim of our research work was to observe the degradation in powder and in solution of different molar mass hyaluronic acid samples, at different temperatures for certain periods. The degradation of the analyzed samples was mainly observed by modifications in molar mass.

Keywords: thermal degradation, hyaluronic acid, BSA, SEC-MALLS

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Authors: Shamim Mushtaq, Moazzam Shahid

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Breast cancer (BC) claims the lives of half a million women every year and is the most common cause of death in the developing world. In 2019, it was estimated that BC alone accounts for 15% of all cancer deaths in younger women (aged < 45 years old) with advanced-stage lung metastasis. According to the World Health Organization & International Union against Cancer, in Asia, a high number of cancer-related deaths will be observed in 2020, whereas the burden will be reduced in Western countries due to awareness about the disease, better health facilities and advanced treatments. In the last 15 years, it has been reported that the incidence of BC has increased by 1.1% among Asian compared to the US population from 2003 to 2012. To date, several BC biological subtypes have been reported so far, which are associated with different treatment responses. The heterogeneity and diversity of BC reflected these different subtypes, including Luminal A (23.7% prevalence) and B (38.8% prevalence) that have pathological estrogen receptor (ER+)-positive tumors, the human epidermal growth factor receptor 2 (HER2) (11.2% prevalence) and triple-negative breast cancer (TNBC) (25% prevalence). According to Shaukat Khanum Memorial Cancer Hospital and Research Centre – Pakistan, ten years of data showed that among 636 BC patients, 30.5% had TNBC who were <40 years of age, which is an extremely alarming situation. Therefore, there is a dire need to explore and develop therapeutic targets for the treatment of early TNBC. Since the last decade, unfortunately, there has been little success in understanding the complexity of TNBC and in discovering new biological therapeutic targets. However, conventional chemotherapy is the only choice of treatment for TNBC patients. Many investigators revealed advances in multi-omics (multiple "omes", e.g., genome, proteome, transcriptome, epigenome, and microbiome) which were later identified as actionable targets and increased prevalence in TNBC patients. However, various drugs have been identified so far which are related to a particular diagnostic and prognostic biomarker. For example, Epidermal growth factor receptor ( EGFR or ErbB-1), HER-2/neu (ErbB-2), HER-3 (ErbB-3), and HER-4 (ErbB-4). Protein Transglin-2 (TAGLN 2 ) and Profilins-1 (Pfn-1 ) are the ubiquitously expressed large family of proteins present in all eukaryotes, enabling actin cytoskeletal reorganization. It is known that the oncogenic transformation of cells is accompanied by alteration in the actin cytoskeleton. There are causal connections between altered expression of actin cytoskeletal regulators and cancer progression. Our case-control study identified TAGLN-2 and Pfn-1 proteins in TNBC blood by mass spectrometry. Both TAGLN-2 and Pfn-1 proteins are differentially expressed in early and advanced stages of TNBS patients, which could be potential predictors or therapeutic targets for TNBC.

Keywords: TNBC, blood biomarkers, mass spectrometry, qPCR, ELISA

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Authors: Amna Abdalla Mohammed Khalid, Pietro Parisse, Silvia Onesti, Loredana Casalis

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Basic cellular processes as DNA replication are crucial to cell life. Understanding at the molecular level the mechanisms that govern DNA replication in proliferating cells is fundamental to understand disease connected to genomic instabilities, as a genetic disease and cancer. A key step for DNA replication to take place, is unwinding the DNA double helix and this carried out by proteins called helicases. The archaeal MCM (minichromosome maintenance) complex from Methanothermobacter thermautotrophicus have being studied using Atomic Force Microscopy (AFM), imaging in air and liquid (Physiological environment). The accurate analysis of AFM topographic images allowed to understand the static conformations as well the interaction dynamic of MCM and DNA double helix in the present of ATP.

Keywords: DNA, protein-DNA interaction, MCM (mini chromosome manteinance) complex, atomic force microscopy (AFM)

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Authors: Minh Huynh, Aaron Heuser, Luke Patterson, Chris Zhang, Mason Miller, Daniel Wang, Sandeep Shetty, Mike Trinh, Abigail Miller, Adaeze Enekwechi, Tenille Daniels, Lu Huynh

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Machine learning and text analytics have been used to analyze child abuse, cyberbullying, domestic abuse and domestic violence, and hate speech. However, to the authors’ knowledge, no research to date has used these methods to study elder abuse in nursing homes or skilled nursing facilities from field inspection reports. We used machine learning and text analytics methods to analyze 356,000 inspection reports, which have been extracted from CMS Form-2567 field inspections of US nursing homes and skilled nursing facilities between 2016 and 2021. Our algorithm detected occurrences of the various types of abuse, including physical abuse, psychological abuse, verbal abuse, sexual abuse, and passive and active neglect. For example, to detect physical abuse, our algorithms search for combinations or phrases and words suggesting willful infliction of damage (hitting, pinching or burning, tethering, tying), or consciously ignoring an emergency. To detect occurrences of elder neglect, our algorithm looks for combinations or phrases and words suggesting both passive neglect (neglecting vital needs, allowing malnutrition and dehydration, allowing decubiti, deprivation of information, limitation of freedom, negligence toward safety precautions) and active neglect (intimidation and name-calling, tying the victim up to prevent falls without consent, consciously ignoring an emergency, not calling a physician in spite of indication, stopping important treatments, failure to provide essential care, deprivation of nourishment, leaving a person alone for an inappropriate amount of time, excessive demands in a situation of care). We further compare the prevalence of abuse before and after Covid-19 related restrictions on nursing home visits. We also identified the facilities with the most number of cases of abuse with no abuse facilities within a 25-mile radius as most likely candidates for additional inspections. We also built an interactive display to visualize the location of these facilities.

Keywords: machine learning, text analytics, elder abuse, elder neglect, nursing home abuse

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Authors: Amruta D. S. Pathare, Indira Hinduja, Kusum Zaveri

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Implantation failure (IF) even after the good-quality embryo transfer (ET) in the physiologically normal endometrium is the main obstacle in in vitro fertilization (IVF). Various microarray studies have been performed worldwide to elucidate the genes requisite for endometrial receptivity. These studies have included the population based on different phases of menstrual cycle during natural cycle and stimulated cycle in normal fertile women. Additionally, the literature is also available in recurrent implantation failure patients versus oocyte donors in natural cycle. However, for the first time, we aim to study the genomics of endometrial receptivity in IF patients under controlled ovarian stimulation (COS) during which ET is generally practised in IVF. Endometrial gene expression profiling in IF patients (n=10) and oocyte donors (n=8) were compared during window of implantation under COS by whole genome microarray (using Illumina platform). Enrichment analysis of microarray data was performed to determine dys-regulated biological functions and pathways using Database for Annotation, Visualization and Integrated Discovery, v6.8 (DAVID). The enrichment mapping was performed with the help of Cytoscape software. Microarray results were validated by real-time PCR. Localization of genes related to immune response (Progestagen-Associated Endometrial Protein (PAEP), Leukaemia Inhibitory Factor (LIF), Interleukin-6 Signal Transducer (IL6ST) was detected by immunohistochemistry. The study revealed 418 genes downregulated and 519 genes upregulated in IF patients compared to healthy fertile controls. The gene ontology, pathway analysis and enrichment mapping revealed significant downregulation in activation and regulation of immune and inflammation response in IF patients under COS. The lower expression of Progestagen Associated Endometrial Protein (PAEP), Leukemia Inhibitory Factor (LIF) and Interleukin 6 Signal Transducer (IL6ST) in cases compared to controls by real time and immunohistochemistry suggests the functional importance of these genes. The study was proved useful to uncover the probable reason of implantation failure being imbalance of immune and inflammatory regulation in our group of subjects. Based on the present study findings, a panel of significant dysregulated genes related to immune and inflammatory pathways needs to be further substantiated in larger cohort in natural as well as stimulated cycle. Upon which these genes could be screened in IF patients during window of implantation (WOI) before going for embryo transfer or any other immunological treatment. This would help to estimate the regulation of specific immune response during WOI in a patient. The appropriate treatment of either activation of immune response or suppression of immune response can be then attempted in IF patients to enhance the receptivity of endometrium.

Keywords: endometrial receptivity, immune and inflammatory response, gene expression microarray, window of implantation

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Authors: Yousuf Al Suleimani, Ahmed Al Mahruqi

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A typical cannabinoid O-1602 induces vasorelaxation and activates the orphan G protein-coupled receptor GPR55 in human endothelial cells. The aim of this study is to characterize the mechanisms of endothelium-independent relaxation of O-1602 in the rat small mesenteric artery using wire myograph. In endothelium-denuded vessels, O-1602 partially produced concentration-dependent vasorelaxation. In vessels depleted of intracellular Ca2+ (by EGTA and methoxamine), CaCl2 produced concentration-dependent contraction. Preincubation with O-1602 (at 10 µM and 30 µM) abolished the contractile responses (P<0.01). The putative antagonist at novel “endothelial anandamide receptor” O-1918 (10 µM) significantly reversed the inhibitory effect of O-1602 on CaCl2-induced vasoconstriction. It is likely that the mechanism of endothelium-independent vasorelaxation to O-1602 is mediated by interfering with Ca2+ entry via an O-1918-sensitive pathway.

Keywords: O-1602, endothelium, vasorelaxation, calcium

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Authors: Mustafa M. Donma, Orkide Donma

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Zinc is a vital element required for growth and development. This fact makes zinc important, particularly for children. It maintains normal cellular structure and functions. This essential element appears to have protective effects against coronary artery disease and cardiomyopathy. Higher serum zinc levels are associated with lower risk of cardiovascular diseases (CVDs). There is a significant association between low serum zinc levels and heart failure. Zinc may be a potential biomarker of cardiovascular health. High sensitive cardiac troponin T (hs-cTnT) and cardiac myosin binding protein C (cMyBP-C) are new generation markers used for prediagnosis, diagnosis, and prognosis of CVDs. The aim of this study is to determine zinc as well as new generation cardiac markers profiles in children with normal body mass index (N-BMI), obese (OB), morbid obese (MO) children, and children with metabolic syndrome (MetS) findings. The association among them will also be investigated. Four study groups were constituted. The study protocol was approved by the institutional Ethics Committee of Tekirdag Namik Kemal University. Parents of the participants filled informed consent forms to participate in the study. Group 1 is composed of 44 children with N-BMI. Group 2 and Group 3 comprised 43 OB and 45 MO children, respectively. Forty-five MO children with MetS findings were included in Group 4. World Health Organization age- and sex-adjusted BMI percentile tables were used to constitute groups. These values were 15-85, 95-99, and above 99 for N-BMI, OB, and MO, respectively. Criteria for MetS findings were determined. Routine biochemical analyses, including zinc, were performed. High sensitive-cTnT and cMyBP-C concentrations were measured by kits based on enzyme-linked immunosorbent assay principle. Appropriate statistical tests within the scope of SPSS were used for the evaluation of the study data. p<0.05 was accepted as statistically significant. Four groups were matched for age and gender. Decreased zinc concentrations were measured in Groups 2, 3, and 4 compared to Group 1. Groups did not differ from one another in terms of hs-cTnT. There were statistically significant differences between cMyBP-C levels of MetS group and N-BMI as well as OB groups. There was an increasing trend going from N-BMI group to MetS group. There were statistically significant negative correlations between zinc and hs-cTnT as well as cMyBP-C concentrations in MetS group. In conclusion, inverse correlations detected between zinc and new generation cardiac markers (hs-TnT and cMyBP-C) have pointed out that decreased levels of this physiologically essential trace element accompany increased levels of hs-cTnT as well as cMyBP-C in children with MetS. This finding emphasizes that both zinc and these new generation cardiac markers may be evaluated as biomarkers of cardiovascular health during severe childhood obesity precipitated with MetS findings and also suggested as the messengers of the future risk in the adulthood periods of children with MetS.

Keywords: cardiac myosin binding protein-C, cardiovascular diseases, children, high sensitive cardiac troponin T, obesity

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Authors: Labiba Zerari Bourafa, Djebar Mohamed Reda, Berrebah Houria, Khadri Sihem, Chiheb Linda

Abstract:

The study of the effect of metal dust (pollutants) was performed on higher plant white beans Phaseolus vulgaris; the experience took place in cellular toxicology laboratory (in vitro culture). The seeds of the bean Phaseolus vulgaris are cultured in a metal contaminated dust medium (a single treatment by different increasing doses), at a rate of 10 seeds per box, for 10 days. The measurement of morpho-metric parameters is performed during the first 96 hours that follow the germination; while the dosage of the proline, the protein content and histological sections are formed on the tenth day (240 h). All morpho-metric and biochemical parameters measured were highly disturbed by metal dust; histological sections confirm this disurbance.

Keywords: conductive fabrics, metal dust, osmoticums, roots, Phaseolus vulgaris

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Authors: Ryan D. Martinus

Abstract:

Inflammation of the endothelium is an important process leading to diabetic atherosclerosis. However, the molecular mechanisms by which diabetes contributes to endothelial inflammation remain to be established. Using In-vitro cultured Human cells and Hsp60 specific ELISA assays, we show that Hsp60 is not only induced in Human monocyte cells under hyperglycaemic conditions but that the Hsp60 is also secreted from these cells. Furthermore, we also demonstrate that the Hsp60 secreted from these monocyte cells is also able to activate Toll-like receptor-4 (TLR4) from Human endothelial cells. This suggests that a potential link may exist between the hyperglycaemia-induced expression of Hsp60 in monocyte cells and vascular inflammation. Circulating levels of Hsp60 due to mitochondrial stress in diabetes patients could, therefore, be an important modulator of inflammation in endothelial cells and thus contribute to the increased incidences of atherosclerosis in diabetes mellitus.

Keywords: mitochondria, Hsp60, inflammation, diabetes mellitus

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Authors: Esther Van Andel, Stefanie C. Lange, Maarten M. J. Smulders, Han Zuilhof

Abstract:

False outcomes of diagnostic tests are a major concern in medical health care. To improve the reliability of surface-based diagnostic tests, it is of crucial importance to diminish background signals that arise from the non-specific binding of biomolecules, a process called fouling. The aim is to create surfaces that repel all biomolecules except the molecule of interest. This can be achieved by incorporating antifouling protein repellent coatings in between the sensor surface and it’s recognition elements (e.g. antibodies, sugars, aptamers). Zwitterionic polymer brushes are considered excellent antifouling materials, however, to be able to bind the molecule of interest, the polymer brushes have to be functionalized and so far this was only achieved at the expense of either antifouling or binding capacity. To overcome this limitation, we combined both features into one single monomer: a zwitterionic sulfobetaine, ensuring antifouling capabilities, equipped with a clickable azide moiety which allows for further functionalization. By copolymerizing this monomer together with a standard sulfobetaine, the number of azides (and with that the number of recognition elements) can be tuned depending on the application. First, the clickable azido-monomer was synthesized and characterized, followed by copolymerizing this monomer to yield functionalizable antifouling brushes. The brushes were fully characterized using surface characterization techniques like XPS, contact angle measurements, G-ATR-FTIR and XRR. As a proof of principle, the brushes were subsequently functionalized with biotin via strain-promoted alkyne azide click reactions, which yielded a fully zwitterionic biotin-containing 3D-functionalized coating. The sensing capacity was evaluated by reflectometry using avidin and fibrinogen containing protein solutions. The surfaces showed excellent antifouling properties as illustrated by the complete absence of non-specific fibrinogen binding, while at the same time clear responses were seen for the specific binding of avidin. A great increase in signal-to-noise ratio was observed, even when the amount of functional groups was lowered to 1%, compared to traditional modification of sulfobetaine brushes that rely on a 2D-approach in which only the top-layer can be functionalized. This study was performed on stoichiometric silicon nitride surfaces for future microring resonator based assays, however, this methodology can be transferred to other biosensor platforms which are currently being investigated. The approach presented herein enables a highly efficient strategy for selective binding with retained antifouling properties for improved signal-to-noise ratios in binding assays. The number of recognition units can be adjusted to a specific need, e.g. depending on the size of the analyte to be bound, widening the scope of these functionalizable surface coatings.

Keywords: antifouling, signal-to-noise ratio, surface functionalization, zwitterionic polymer brushes

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