Search results for: PadR family protein

Authors: Reshma Parvin Nuri, Heather Michelle Aldersey, Setareh Ghahari

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Objectives: Worldwide, it is well known that taking care of children with disabilities (CWD) can have a significant impact on the entire family unit. Over the last few decades, an increased number of studies have been conducted on families of CWD in higher income countries, and much of this research has identified family needs and strategies to meet those needs. However, family needs are incredibly under-studied in developing countries. Therefore, the aims of this study were to: (a) explore the needs of families of children with cerebral palsy (CP) in Bangladesh; (b) investigate how some of the family needs have been met and (c) identify the sources of supports that might help the families to meet their needs in the future. Methods: A face to face, semi-structured in-depth interview was conducted with 20 family members (12 mothers, 4 fathers, 1 sister, 2 grandmothers, and 1 aunt) who visited the Centre for the Rehabilitation of the Paralysed (CRP), Bangladesh between June and August 2016. Constant comparison method of grounded theory approach within the broader spectrum of qualitative study was used to analyze the data. Results: Participants identified five categories of needs: (a) financial needs, (b) access to disability-related services, (c) family and community cohesion, (d) informational needs, and (e) emotional needs. Participants overwhelmingly reported that financial need is their greatest family need. Participants noted that families encountered additional financial expenses for a child with CP, beyond what they would typically pay for their other children. Participants were seeing education as their non-primary need as they had no hope that their children would be physically able to go to school. Some participants also shared their needs for social inclusion and participation and receiving emotional support. Participants further expressed needs to receive information related to the child’s health condition and availability/accessibility of governmental support programs. Besides unmet needs, participants also highlighted that some of their needs have been met through formal and informal support systems. Formal support systems were mainly institution-based and run by non-governmental organizations, whereas participants identified informal support coming from family, friends and community members. Participants overwhelmingly reported that they receive little to no support from the government. However, participants identified the government as the key stakeholder who can play vital role in meeting their unmet needs. Conclusions: In the next phase of this research, the plan is to understand how the Government of the People’s Republic of Bangladesh is working to meet the needs of families of CWD. There is also need for further study on needs of families of children with conditions other than CP and those who live in the community and do not have access to the CRP Services. There is clear need to investigate ways to enable children with CP have better access to education in Bangladesh.

Keywords: Bangladesh, children with cerebral palsy, family needs, support

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Authors: E. Obreque-Slier, V. Contreras-Cortez, R. López-Solís

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Astringency is a drying, roughing, and sometimes puckering sensation that is experienced on the various oral surfaces during or immediately after tasting foods. This sensation has been closely related to the interaction and precipitation between salivary proteins and polyphenols, specifically flavanols or proanthocyanidins. In addition, the type and concentration of proanthocyanidin influences significantly the intensity of the astringency and consequently the protein/proanthocyanidin interaction. However, most of the studies are based on the interaction between saliva and highly complex polyphenols, without considering the effect of monomeric proanthoancyanidins present in different foods. The aim of this study was to evaluate the effect of different monomeric proanthocyanidins on the diffusion and precipitation of salivary proteins. Thus, solutions of catechin, epicatechin, epigallocatechin and gallocatechin (0, 2.0, 4.0, 6.0, 8.0 and 10 mg/mL) were mixed with human saliva (1: 1 v/v). After incubation for 5 min at room temperature, 15 µL aliquots of each mix were dotted on a cellulose membrane and allowed to dry spontaneously at room temperature. The membrane was fixed, rinsed and stained for proteins with Coomassie blue. After exhaustive washing in 7% acetic acid, the membrane was rinsed once in distilled water and dried under a heat lamp. Both diffusion area and stain intensity of the protein spots were semiqualitative estimates for protein-tannin interaction (diffusion test). The rest of the whole saliva-phenol solution mixtures of the diffusion assay were centrifuged, and 15-μL aliquots from each of the supernatants were dotted on a cellulose membrane. The membrane was processed for protein staining as indicated above. The blue-stained area of protein distribution corresponding to each of the extract dilution-saliva mixtures was quantified by Image J 1.45 software. Each of the assays was performed at least three times. Initially, salivary proteins display a biphasic distribution on cellulose membranes, that is, when aliquots of saliva are placed on absorbing cellulose membranes, and free diffusion of saliva is allowed to occur, a non-diffusible protein fraction becomes surrounded by highly diffusible salivary proteins. In effect, once diffusion has ended, a protein-binding dye shows an intense blue-stained roughly circular area close to the spotting site (non-diffusible fraction) (NDF) which becomes surrounded by a weaker blue-stained outer band (diffusible fraction) (DF). Likewise, the diffusion test showed that epicatechin caused the complete disappearance of DF from saliva with 2 mg/mL. Also, epigallocatechin and gallocatechin caused a similar effect with 4 mg/mL, while catechin generated the same effect at 8 mg/mL. In the precipitation test, the use of epicatechin and gallocatechin generated evident precipitates at the bottom of the Eppendorf tubes. In summary, the flavanol type differentially affects the diffusion and precipitation of saliva, which would affect the sensation of astringency perceived by consumers.

Keywords: astringency, polyphenols, tannins, tannin-protein interaction

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Authors: Alshutwi Sitah

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Many countries around the world are struggling to maintain an adequate number of nurses. Inadequate nursing staffing could compromise the quality of patient care. Among many factors that contribute to registered nurses (RN) turnover, the influence of work–family conflict (WFC) has gained little attention. WFC was found to be significantly associated with increased turnover intention (TI) among employees. Furthermore, WFC has been linked to a number of negative consequences, including lower job satisfaction and organizational commitment, sleep insufficiency, insomnia symptoms, obesity, cardiovascular diseases, sleep insufficiency, and high cholesterol. In an effort to find strategies to manage the consequences of WFC, many behavioral, psychological, and career scholars have focused on the role of supervisor support. Family Supportive Supervisor Behaviors (FSSB) has been found to be a promising approach contributing to the reduction of TI in employees’ experiencing WFC. Despite the importance of work–family issues and the influence of FSSB, limited studies have been conducted among the nursing population and none were found that included a sample from Saudi Arabia. Therefore, the main Purpose of this study was to evaluate the influence of FSSB on the relationship among WFC, Stress, and TI in Saudi Arabian registered nurses. Method: A cross-sectional study. Sample: Convenience sampling; 113 Saudi female nurse. Result: Fifty percent of nurses intended to leave their workplace, 68 % of nurses reported having a conflict between work and family, and 44% reported having a high level of stress. A significant positive correlation was found between WFC and TI (r= .43, P < 0.01). A negative correlation was found between FSSB and TI (r= -.53, P < 0.01). Both WFC and stress were associated with TI; however, these associations were buffered (weaken), when nurses had higher FSSB. Conclusion: The FSSB could be seen as a tool to help married, female nurses to demonstrate their professional role without compromising their family responsibilities. Nurses’ turnover is a complex issue that may require multiple prevention strategies; however, enhancing FSSB could be a key resource for maintaining a positive workplace environment and reducing TI.

Keywords: turnover intention, work-family conflict, supervisor support, nursing retention

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Authors: Shahram Naghizadeh Raeisi, Ali Alghooneh, Seyed Jalal Razavi Zahedkolaei

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Herein, a shelf stable (no refrigeration required) UHT processed, aseptically packaged whey protein drink was formulated by using a new strategy in microparticulate process. Applying thermal and two-dimensional mechanical treatments simultaneously, a modified protein (MWPC-80) was produced. Then the physical, thermal and thermodynamic properties of MWPC-80 were assessed using particle size analysis, dynamic temperature sweep (DTS), and differential scanning calorimetric (DSC) tests. Finally, using MWPC-80, a new RTD beverage was formulated, and shelf stability was assessed for three months at ambient temperature (25 °C). Non-isothermal dynamic temperature sweep was performed, and the results were analyzed by a combination of classic rate equation, Arrhenius equation, and time-temperature relationship. Generally, results showed that temperature dependency of the modified sample was significantly (Pvalue<0.05) less than the control one contained WPC-80. The changes in elastic modulus of the MWPC did not show any critical point at all the processed stages, whereas, the control sample showed two critical points during heating (82.5 °C) and cooling (71.10 °C) stages. Thermal properties of samples (WPC-80 & MWPC-80) were assessed using DSC with 4 °C /min heating speed at 20-90 °C heating range. Results did not show any thermal peak in MWPC DSC curve, which suggested high thermal resistance. On the other hands, WPC-80 sample showed a significant thermal peak with thermodynamic properties of ∆G:942.52 Kj/mol ∆H:857.04 Kj/mole and ∆S:-1.22Kj/mole°K. Dynamic light scattering was performed and results showed 0.7 µm and 15 nm average particle size for MWPC-80 and WPC-80 samples, respectively. Moreover, particle size distribution of MWPC-80 and WPC-80 were Gaussian-Lutresian and normal, respectively. After verification of microparticulation process by DTS, PSD and DSC analyses, a 10% why protein beverage (10% w/w/ MWPC-80, 0.6% w/w vanilla flavoring agent, 0.1% masking flavor, 0.05% stevia natural sweetener and 0.25% citrate buffer) was formulated and UHT treatment was performed at 137 °C and 4 s. Shelf life study did not show any jellification or precipitation of MWPC-80 contained beverage during three months storage at ambient temperature, whereas, WPC-80 contained beverage showed significant precipitation and jellification after thermal processing, even at 3% w/w concentration. Consumer knowledge on nutritional advantages of whey protein increased the request for using this protein in different food systems especially RTD beverages. These results could make a huge difference in this industry.

Keywords: high protein whey beverage, micropartiqulation, two-dimentional mechanical treatments, thermodynamic properties

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Authors: Sirilak Kongkaew, Pathumwadee Yodmanee, Nopporn Kaiyawet, Arthitaya Meeprasert, Thanyada Rungrotmongkol, Toshikatsu Kaburaki, Hiroshi Noguchi, Fujio Takeuch, Nawee Kungwan, Supot Hannongbua

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Behçet’s disease (BD) is a genetic autoimmune expressed by multisystemic inflammatory disorder mostly occurred at the skin, joints, gastrointestinal tract, and genitalia, including ocular, oral, genital, and central nervous systems. Most BD patients in Japan and Korea were strongly indicated by the genetic factor namely HLA-B*51 (especially, HLA-B*51:01) marker in HMC class I, while HLA-A*26:01 allele has been detected from the BD patients in Greek, Japan, and Taiwan. To understand the selective binding of the MICA-TM peptide towards the HLAs associated with BD, the molecular dynamics simulations were applied on the four HLA alleles (B*51:01, B*35:01, A*26:01, and A*11:01) in complex with such peptide. As a result, the key residues in the binding groove of HLA protein which play an important role in the MICA-TM peptide binding and stabilization were revealed. The Van der Waals force was found to be the main protein-protein interaction. Based on the binding free energy prediction by MM/PBSA method, the MICA-TM peptide interacted stronger to the HLA alleles associated to BD in the identical class by 7-12 kcal/mol. The obtained results from the present study could help to differentiate the HLA alleles and explain a source of Behçet’s disease.

Keywords: Behçet’s disease, MD simulations, HMC class I, autoimmune

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Authors: Salma Akter, Mohammad M. Haque, Lawrence Akwetey

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An empirical research design will analyze different factors/predictors children use to influence their parents in the family buying decision process in the unexplored area of British Bangladeshi children in the United Kingdom. The proposed conceptual model of factors- buying decision making process will be tested by the Structure Equation Model. A structured Questionnaire and secondary sources will employ to collect data and analyse and measure the validity by Statistical tools (SPSS) and Microsoft Excel. The Contemporary research aims to use the deductive approach developing the research questions and testing the hypothesis to identify the impact of different strategies British Bangladeshi children used to influence their parents in the family buying decision which was overlooked in the previous research.

Keywords: British Bangladeshi children, buying decision process, children influence, influential factors

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Authors: Hani M. M. Alothaid, Louise Robson, Richmond Muimo

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This study will investigate cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) dependent calcineurin (Cn), known as protein phosphatase 2 B (PP2B) as well, regulation of chloride ion (Cl⁻) secretion and the release of pro-inflammatory molecules in immune cells such as cytokines. THP-1-derived monocytes, primary human monocytes and the bronchial epithelial cell line (16HBE14o-) were used in this study. The 16HBE14o- cells were chosen as positive control. Hence, to further confirm the expression of cystic fibrosis transmembrane conductance regulator (CFTR), calcium binding protein (S100A10), annexin A2 (AnxA2) and calcineurin A subunit (CnA) in all three cell types, cell lysate was probed against corresponding primary antibodies by immunoblotting. Western blot analyses show the expression of CFTR, AnxA2, CnA and S100A10 in THP-1-derived monocytes and primary human monocytes. In conclusion, CFTR, S100A10, CnA and AnxA2 are expressed in THP-1-derived monocytes and primary human monocytes and regulate Cl⁻ secretion. Also, they may play a role in the pro-inflammatory molecules release. The ongoing work will confirm interaction between these proteins in the cell lines.

Keywords: annexin A2, calcineurin, CFTR, chloride, monocytes, pro-inflammatory molecules, S100A10

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Authors: Mohamed Yusuf

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Aim: The growth promoting the effect of the blue-green filamentous alga Spirulina platensis (SP) was observed on meat type Japanese quail with antibiotic growth promoter alternative and immune enhancing power. Materials and Methods: This study was conducted on 180 Japanese quail chicks for 4 weeks to find out the effect of diet type (vegetarian protein diet [VPD] and fish meal protein diet [FMPD])- Spirulina dose interaction (1 or 2 g/kg diet) on growth performance, gut microbiota, and sensory meat quality of growing Japanese quails (1-5 weeks old). Results: Data revealed improvement (p<0.05) of weight gain, feed conversion ratio, and European efficiency index due to 1, 2 g (SP)/kg VPD, and 2 g (SP)/kg FMPD, respectively. There was a significant decrease of ileum mean pH value by 1 g(SP)/kg VPD. Concerning gut microbiota, there was a trend toward an increase in Lactobacilli count in both 1; 2 g (SP)/kgVPD and 2 g (SP)/kg FMPD. It was concluded that 1 or 2 g (SP)/kg vegetarian diet may enhance parameters of performance without obvious effect on both meat quality and gut microbiota. Moreover, 1 and/or 2 g (SP) may not be invited to share fishmeal based diet for growing Japanese quails. Conclusion: Using of SP will support the profitable production of Japanese quails fed vegetable protein diet.

Keywords: isocaloric, isonitrogenous, meat quality, performances, quails, spirulina, spirulina

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Authors: Khaled S. Al Salhen

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Aldehyde oxidase (AO) and xanthine oxidoreductase (XOR) catalyze the oxidation of many different N-heterocyclic compounds as well as aliphatic and aromatic aldehydes to their corresponding lactam and carboxylic acids respectively. The present study examines the oxidation of dimethylamino-cinnamaldehyde (DMAC), vanillin and phenanthridine by AO and xanthine by XOR from Drosophila cytosol. Therefore, the results obtained in the present study showed the DMAC, vanillin and phenanthridine substrates used were found to be good substrates of Drosophila AO and xanthine is the preferred substrate for Drosophila XOR. Km values of AO substrates were observed with DMAC (50±5.4 µM), phenanthridine (80±9.1 µM) and vanillin (303±11.7 µM) respectively for Drosophila cytosol. The Km values for DMAC and phenanthridine were ~6 and ~4 fold lower than that for vanillin as a substrate. The Km for XOR with xanthine using NAD+ as an electron acceptor was 27±4.1 µM. Relatively low Vmax values were obtained with phenanthridine (1.78±0.38 nmol/min/mg protein) and DMAC (1.80±0.35 nmol/min/mg protein). The highest Vmax was obtained from Drosophila cytosol with vanillin (7.58±2.11 nmol/min/mg protein). It is concluded these results that AO and XOR in Drosophila were able to catalyse the biotransformation of numerous substrates of the well-characterised mammalian AO and XOR. The kinetic parameters have indicated that the activity of AO of Drosophila may be a significant factor the oxidation of aromatic aldehyde compounds.

Keywords: aldehyde oxidase, xanthine oxidoreductase, dimethylamino-cinnamaldehyde, vanillin, phenanthridine, Drosophila melanogaster

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Authors: Hanna Gulema, Meaza Demissie, Alemayehu Worku, Tesfaye Assebe Yadeta, Yemane Berhane

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Background: In patriarchal societies, female caregivers decide on food allocation within a family based on prevailing gender and age norms, which may lead to inequality that does not favor young adolescent girls. This study evaluated the effect of a community-based social norm intervention involving female caregivers in West Hararghe, Ethiopia. The intervention was engaging female caregivers along with other adult influential community members to deliberate and act on food allocation social norms in a process referred to as Social Analysis and Action (SAA). Method: We used data from a large quasi-experimental study to compare family eating practices between those who participated in the Social Analyses and Action intervention and those who did not. The respondents were female caregivers in households with young adolescent girls (ages 13 and 14 years). The study’s outcome was the practice of family eating together from the same dish. The difference in difference (DID) analysis with the Mixed effect logistic regression model was used to examine the effect of the intervention. Result: The results showed improved family eating practices in both groups, but the improvement was greater in the intervention group. The DID analysis showed an 11.99 percentage points greater improvement in the intervention arm than in the control arm. The mixed-effect regression produced an adjusted odds ratio of 2.08 (95% CI [1.06–4.09]) after controlling selected covariates, p-value 0.033. Conclusions: The involvement of influential adult community members significantly improves the family practice of eating together in households where adolescent girls are present in our study. The intervention has great potential to minimize household food allocation inequalities and thus improve the nutritional status of young adolescents. Further studies are necessary to evaluate the effectiveness of the intervention in different social norm contexts to formulate policy and guidelines for scale-up.

Keywords: family eating practice, social norm intervention, adolescence girls, caregiver

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Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki

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Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.

Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism

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Authors: Ali Christian Chinedu, Asogwa Vincent Chidindu, Ejiofor Toochukwu Eleazar, Okadi Ashagwu Ojang

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The increasing rate of Boko Haram insurgency, farmer-herder clashes, and kidnapping in Nigeria has resulted in food shortages and high cost of protein sources like beef and fish. This challenge could be curbed with the production of edible insects, which contain several nutritional benefits like calories, protein, fat, vitamins, and minerals, depending on their species, metamorphic stage, and diet. Unfortunately, the benefits and competencies in producing, preserving, and marketing edible insects are still unknown to the public, including prospective farmers in Nigeria. Hence, this study determined teachers’ perception of integrating edible insects into the Animal Husbandry Curriculum in Senior Secondary Schools in Nigeria to equip the future generation with the relevant competencies for alternative sustainable protein supply. The study was carried out in Enugu State, Nigeria. The participants for the study comprised 162 agricultural science teachers. A questionnaire titled: Edible Insects Integration in Animal Husbandry Curriculum Questionnaire (EIIAHCQ) was used to collect data using a descriptive survey research design. We conducted data collection with the help of six research assistants. The study identified 11 objectives, 11 contents, 10 teaching methods, and 9 evaluation methods that could be integrated into the existing curriculum of animal husbandry in Nigeria. Among others, the Ministry of Education should integrate the finding of this study into the curriculum of Animal Husbandry in Nigeria to enhance the protein supply and curb food insecurity now and in the future.

Keywords: animal husbandry curriculum, edible insects, entomophagy, integration, secondary school, Nigeria

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Authors: Xian-Peng Jiang, Catherine C. Baucom, Toby Jiang, Robert L. Elliott

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HLA-G binds to the inhibitory receptors of uterine NK cells and plays an important role in protection of fetal cells from maternal NK lysis. HLA-G also mediates tumor escape, but the immunosuppressive role is often neglected. These studies have focused on the examination of HLA-G expression in human breast carcinoma and HLA-G immunosuppressive role in NK cytolysis. We examined HLA-G expression in breast cell lines by real time PCR, ELISA and immunofluorescent staining. We treated the breast cancer cell lines with anti-human HLA-G antibody or progesterone. Then, NK cytolysis was measured by using MTT assay. We find that breast carcinoma cell lines increase the expression of HLA-G mRNA and protein, compared to normal cells. Blocking HLA-G of the breast cancer cells by the antibody increases NK cytolysis. Progesterone upregulates HLA-G mRNA and protein of human breast cancer cell lines. The increased HLA-G expression suppresses NK cytolysis. In summary, human breast carcinoma overexpress HLA-G immunosuppressive molecules. Blocking HLA-G protein by antibody improves NK cytolysis. In contrast, upregulation of HLA-G expression by progesterone impairs NK cytolytic function. Thus, HLA-G is a new immunosuppressive checkpoint and potential cancer immunotherapeutic target.

Keywords: HLA-G, Breast carcinoma, NK cells, Immunosuppressive checkpoint

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Authors: Maneesh Kumar, Roshan Kamal Topno, Manas Ranjan Dikhit, Vahab Ali, Ganesh Chandra Sahoo, Bhawana, Major Madhukar, Rishikesh Kumar, Krishna Pandey, Pradeep Das

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Chandipura vesiculovirus is an emerging (-) ssRNA viral entity belonging to the genus Vesiculovirus of the family Rhabdoviridae, associated with fatal encephalitis in tropical regions. The multi-functionally active viral RNA-dependent RNA polymerase (vRdRp) that has been incorporated with conserved amino acid residues in the pathogens, assigned to synthesize distinct viral polypeptides. The lack of proofreading ability of the vRdRp produces many mutated variants. Here, we have performed the evolutionary analysis of 20 viral protein sequences of vRdRp of different strains of Chandipura vesiculovirus along with other viral species from genus Vesiculovirus inferred in MEGA6.06, employing the Neighbour-Joining method. The p-distance algorithmic method has been used to calculate the optimum tree which showed the sum of branch length of about 1.436. The percentage of replicate trees in which the associated taxa are clustered together in the bootstrap test (1000 replicates), is shown next to the branches. No mutation was observed in the Indian strains of Chandipura vesiculovirus. In vRdRp, 1230(His) and 1231(Arg) are actively participated in catalysis and, are found conserved in different strains of Chandipura vesiculovirus. Both amino acid residues were also conserved in the other viral species from genus Vesiculovirus. Many isolates exhibited maximum number of mutations in catalytic regions in strains of Chandipura vesiculovirus at position 26(Ser→Ala), 47 (Ser→Ala), 90(Ser→Tyr), 172(Gly→Ile, Val), 172(Ser→Tyr), 387(Asn→Ser), 1301(Thr→Ala), 1330(Ala→Glu), 2015(Phe→Ser) and 2065(Thr→Val) which make them variants under different tropical conditions from where they evolved. The result clarifies the actual concept of RNA evolution using vRdRp to develop as an evolutionary marker. Although, a limited number of vRdRp protein sequence similarities for Chandipura vesiculovirus and other species. This might endow with possibilities to identify the virulence level during viral multiplication in a host.

Keywords: Chandipura, (-) ssRNA, viral RNA-dependent RNA polymerase, neighbour-joining method, p-distance algorithmic, evolutionary marker

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Authors: Gajanan M. Sonwane

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The computational studies on 2-phenazinamines with their protein targets have been carried out to design compounds with potential anticancer activity. This strategy of designing compounds possessing selectivity over specific tyrosine kinase has been achieved through G-QSAR and molecular docking studies. The objective of this research has been to design newer 2-phenazinamine derivatives as Bcr-Abl tyrosine kinase inhibitors by G-QSAR, molecular docking studies followed by wet-lab studies along with evaluation of their anticancer potential. Computational chemistry was done by using VLife MDS 4.3 and Autodock 4.2 followed by wet-lab experiments for synthesizing 2-phenazinamine derivatives. The chemical structures of ligands in 2D were drawn by employing Chemdraw 2D Ultra 8.0 and were converted into 3D. These were optimized by using a semi-empirical method called MOPAC. The protein structure was retrieved from RCSC protein data bank as a PDB file. The binding interactions of protein and ligands were done by using PYMOL. The molecular properties of the designed compounds were predicted in silico by using Osiris property explorer. The parent compound 2-phenazinamine was synthesized by reduction of 2, 4-dinitro-N-phenyl-benzenamine in the presence of tin chloride followed by cyclization in the presence of nitrobenzene and magnesium sulfate. The derivatization at the amino function of 2-phenazinamine was performed by treating parent compound with various aldehydes in the presence of dicyclohexylcarbodiimide (DCC) and urea to afford 2-(2-chlorophenyl)-3-(phenazine-2-yl) thiazolidine-4-one. Synthesized 39 novel derivatives of 2-phenazinamine and performed antioxidant activity, anti antiproliferative on the bulb of onion and anticancer activity on cell line showing significant competition with marked blockbuster drug imatinib.

Keywords: computer-aided drug design, tyrosin kinases, anticancer, docking

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Authors: Simbiat O. Ayilara-Akande, Soji Fakoya

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Quality and yield are always the target of farmers, including mushroom farmers. This study investigated how better Pleurotus pulmonarius can be obtained with the induction of mutagens into the process of spawn production in order to improve both the quality and the yield. Mushroom spawns were treated with ultraviolet radiation (UV) and hydroxylamine hydrochloride (HA) at different exposure times (2, 6, and 10 minutes) and different concentrations (10, 30, and 50Mm), respectively. The treated spawns were used to cultivate mushrooms on five substrates in the family of Gramineae viz: sorghum, rice, bamboo, sugarcane, and corn straws. Matured fruit bodies were harvested after a few weeks, and their parameters were taken and recorded. This study reveals a significant yield increase in mushroom grown on all the substrates when treated with ultraviolet radiation (UV) for 10 minutes and 6 minutes, respectively. Mushroom spawns treated with hydroxylamine hydrochloride showed a negative correlation in the yield with an increased in mutagen concentration. Hence, Ultraviolet light could be employed to enhance the quality and yield of mushroom production.

Keywords: mushroom, protein, mutagens, yield

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Authors: Zhen Cao, Yu Zhu, Junxue Fu

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Immunoassay, i.e., antigen-antibody reaction, is crucial for disease diagnostics. To achieve the adequate signal of the antigen protein detection, a large amount of sample and long incubation time is needed. However, the amount of protein is usually small at the early stage, which makes it difficult to detect. Unlike cells and DNAs, no valid chemical method exists for protein amplification. Thus, an alternative way to improve the signal is through particle manipulation techniques to concentrate proteins, among which dielectrophoresis (DEP) is an effective one. DEP is a technique that concentrates particles to the designated region through a force created by the gradient in a non-uniform electric field. Since DEP force is proportional to the cube of particle size and square of electric field gradient, it is relatively easy to capture larger particles such as cells. For smaller ones like proteins, a super high gradient is then required. In this work, three-dimensional Ag/SiO2 nanorods arrays, fabricated by an easy physical vapor deposition technique called as oblique angle deposition, have been integrated with a DEP device and created the field gradient as high as of 2.6×10²⁴ V²/m³. The nanorods based DEP device is able to enrich bovine serum albumin (BSA) protein by 1800-fold and the rate has reached 180-fold/s when only applying 5 V electric potential. Based on the above nanorods integrated DEP platform, an immunoassay of mouse immunoglobulin G (IgG) proteins has been performed. Briefly, specific antibodies are immobilized onto nanorods, then IgG proteins are concentrated and captured, and finally, the signal from fluorescence-labelled antibodies are detected. The limit of detection (LoD) is measured as 275.3 fg/mL (~1.8 fM), which is a 20,000-fold enhancement compared with identical assays performed on blank glass plates. Further, prostate-specific antigen (PSA), which is a cancer biomarker for diagnosis of prostate cancer after radical prostatectomy, is also quantified with a LoD as low as 2.6 pg/mL. The time to signal saturation has been significantly reduced to one minute. In summary, together with an easy nanorod fabrication and integration method, this nanorods based DEP platform has demonstrated highly sensitive immunoassay performance and thus poses great potentials in applications for early point-of-care diagnostics.

Keywords: dielectrophoresis, immunoassay, oblique angle deposition, protein concentration

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Authors: Hadriana Bansi, Elizabeth Wina, Toto Toharmat

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Acacia villosa is thornless shrub legume which contents high crude protein. However, the utilization of A. villosa as ruminant feed is limited by its secondary compounds. The aim of this article is to find out the maximum of substitution A. villosa in sheep ration. The nutritional evaluation consisted of in vitro two stages, in vivo, and in vitro gas production trials. The secondary compounds of A. villosa also were analyzed. Evaluating digestibility of increasing level of substitution A. villosa replacing Pennisetum purpureum was using in vitro two stages. The substitution of 30% A. villosa was compared to 100% P. purpureum by in vitro gas production technique and in vivo digestibility. The results of two stages in vitro showed that total phenol, condensed tannin, and non-protein amino acid (NPAA) were high. Substitution 15% A. villosa reached the highest digestibility for both dry matter (DM) and crude protein (CP) which were 67% and 86% respectively, but it was shown that DM and CP digestibility of substitution 30% of A. villosa was still high which were 61.82% and 75-67% respectively. The pattern of gas production showed that first 8 hours total gas production substitution of 30% A. villosa was higher than 100% P. purpureum and declined after 10 hours incubation. In vivo trials showed that substitution of 30% A. villosa significantly increased CP intake, CP digestibility, and nitrogen retention. It can be concluded that substitution A. villosa until 30% still gave the good impact even though it has high secondary compounds.

Keywords: Acacia villosa, digestibility, gas production, secondary compounds

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Authors: Debora P. Arce, Flavia J. Krsticevic, Marco R. Bertolaccini, Joaquín Ezpeleta, Estela M. Valle, Sergio D. Ponce, Elizabeth Tapia

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Small Heat Shock Proteins (sHSPs) are low molecular weight chaperones that play an important role during stress response and development in all living organisms. Fruit maturation and oxidative stress can induce sHSP synthesis both in Arabidopsis and tomato plants. RNA-Seq technology is becoming widely used in various transcriptomics studies; however, analyzing and interpreting the RNA-Seq data face serious challenges. In the present work, we de novo assembled the Solanum lycopersicum transcriptome for three different maturation stages (mature green, breaker and red ripe). Differential gene expression analysis was carried out during tomato fruit development. We identified 12 sHSPs differentially expressed that might be involved in breaker and red ripe fruit maturation. Interestingly, these sHSPs have different subcellular localization and suggest a complex regulation of the fruit maturation network process.

Keywords: sHSPs, maturation, tomato, RNA-Seq, assembly

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Authors: Rahamdad Khan, Ijaz Ahmad Khan

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In modern agriculture the herbicides application are considered the most effective and fast in action against all types of weeds. But it’s a fact that the herbicide applicator cannot totally secure the crop plants from the possible herbicide injuries that further leads to several destructive changes in plant biochemistry. For the purpose pots studies were undertaken to test the tolerance order of chickpea against pre- emergence herbicides (Stomp 330 EC- Dual Gold 960 EC) and post- emergence herbicides (Topik 15 WP- Puma Super 75 EW- Isoproturon 500 EW) during 2012-13 and 2013-14. The experimental design was CRD with three replications. Plant height, number of branches plant-1, number of seeds plant-1, nodulation, seed protein contents and other growth related parameters in chickpea were examined during the investigations. The results indicate that all the enquire herbicides gave a significant variation to all recorded parameter of chick pea except nodule fresh and dray weight. Moreover the toxic effect of pre-emergence herbicide on chickpea was found higher as compared to post-emergence herbicides. Minimum chickpea plant height (50.50 cm), number of nodule plant-1 (17.83) and lowest seed protein (14.13 %) was recorded in Stomp 330 EC. Similarly the outmost seeds plant-1 (29.66) and number of nodule plant-1 (21) were found for Puma Super 75 EW. The results further showed that the highest seed protein content (21.75 and 21.15 %) was recorded for control/ untreated and Puma Super 75EW. Taking under concentration the possible negative impact of the herbicides the chemical application must be minimized up to certain extent at which the crop is mostly secure. However chemical weed control has many advantages so we should train our farmer regarding the proper use of agro chemical to minimize the loses in crops while using herbicides.

Keywords: chickpea, herbicides, protein, stomp 330 EC, weed

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Authors: Walter Beck

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Jharkhand is newly constituted 28th State in the eastern part of India which is known for the oldest settlement of the indigenous people. In the State of Jharkhand in which broadly three language family are found namely, Austric, Dravidian, and Indo-European. Ex-Mundari, kharia, Ho Santali come from the Austric Language family. Kurukh, Malto under Dravidian language family and Nagpuri Khorta etc. under Indo-European language family. There are 32 Indigenous Communities identified as Scheduled Tribe in the State of Jharkhand. Santhal, Munda, Kahria, Ho and Oraons are some of the major Tribe of the Jharkhand state. Jharkhand has a Rich Cultural heritage which includes Folk art, folklore, Folk Dance, Folk Music, Folk Songs for which diversity can been seen from place to place, season to season and all traditional Culture and practices. The languages as well as the songs are vulnerable to dominant culture and hence needed to be protected. The collection and documentation of these songs in their natural setting adds significant contribution to the conservation and propagation of the cultural elements. This paper reflects to bring out the Originality of the Collected Songs from remote areas of the plateau of Sothern Jharkhand as a rich intangible Cultural heritage of the Country. The research was done through participatory observation. In this research project more than 100 songs which were never documented before.

Keywords: cultural heritage, India, indigenous people, songs, languages

Procedia PDF Downloads 197

Authors: Mehrnaz Mostafavi

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Recent advances in single-molecule protein identification (ID) and quantification techniques are poised to revolutionize proteomics, enabling researchers to delve into single-cell proteomics and identify low-abundance proteins crucial for biomedical and clinical research. This paper introduces a different approach to single-molecule protein ID and quantification using tri-color amino acid tags and a plasmonic nanopore device. A comprehensive simulator incorporating various physical phenomena was designed to predict and model the device's behavior under diverse experimental conditions, providing insights into its feasibility and limitations. The study employs a whole-proteome single-molecule identification algorithm based on convolutional neural networks, achieving high accuracies (>90%), particularly in challenging conditions (95–97%). To address potential challenges in clinical samples, where post-translational modifications affecting labeling efficiency, the paper evaluates protein identification accuracy under partial labeling conditions. Solid-state nanopores, capable of processing tens of individual proteins per second, are explored as a platform for this method. Unlike techniques relying solely on ion-current measurements, this approach enables parallel readout using high-density nanopore arrays and multi-pixel single-photon sensors. Convolutional neural networks contribute to the method's versatility and robustness, simplifying calibration procedures and potentially allowing protein ID based on partial reads. The study also discusses the efficacy of the approach in real experimental conditions, resolving functionally similar proteins. The theoretical analysis, protein labeler program, finite difference time domain calculation of plasmonic fields, and simulation of nanopore-based optical sensing are detailed in the methods section. The study anticipates further exploration of temporal distributions of protein translocation dwell-times and the impact on convolutional neural network identification accuracy. Overall, the research presents a promising avenue for advancing single-molecule protein identification and quantification with broad applications in proteomics research. The contributions made in methodology, accuracy, robustness, and technological exploration collectively position this work at the forefront of transformative developments in the field.

Keywords: nano proteomics, nanopore-based optical sensing, deep learning, artificial intelligence

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Authors: John W. Travis

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There is an epidemic of couples separating after a child is born into a family, generally with the father leaving emotionally or physically in the first few years after birth. This separation creates high levels of stress for both parents, especially the primary parent, leaving her (or him) less available to the infant for healthy attachment and nurturing. The deterioration of the couple’s bond leaves parents increasingly under-resourced, and the dependent child in a compromised environment, with an increased likelihood of developing an attachment disorder. Objectives: To understand the dynamics of a couple, once the additional and extensive demands of a newborn are added to a nuclear family structure, and to identify effective ways to support all members of the family to thrive. Qualitative studies interviewed men, women, and couples after pregnancy and the early years as a family, regarding key destructive factors, as well as effective tools for the couple to retain a strong bond. In-depth analysis of a few cases, including the author’s own experience, reveal deeper insights about subtle factors, replicated in wider studies. Using a self-assessment survey, many fathers report feeling abandoned, due to the close bond of the mother-baby unit, and in turn, withdrawing themselves, leaving the mother without support and closeness to resource her for the baby. Fathers report various types of abandonment, from his partner to his mother, with whom he did not experience adequate connection as a child. The study identified a key destructive factor to be unrecognized wounding from childhood that was carried into the relationship. The study culminated in the naming of Male Postpartum Abandonment Syndrome (MPAS), describing the epidemic in industrialized cultures with the nuclear family as the primary configuration. A growing family system often collapses without a minimum number of adult caregivers per infant, approximately four per infant (3.87), which allows for proper healing and caretaking. In cases with no additional family or community beyond one or two parents, the layers of abandonment and trauma result in the deterioration of a couple’s relationship and ultimately the family structure. The solution includes engaging community in support of new families. The study identified (and recommends) specific resources to assist couples in recognizing and healing trauma and disconnection at multiple levels. Recommendations include wider awareness and availability of resources for healing childhood wounds and greater community-building efforts to support couples for the whole family to thrive.

Keywords: abandonment, attachment, community building, family and marital functioning, healing childhood wounds, infant wellness, intimacy, marital satisfaction, relationship quality, relationship satisfaction

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Authors: Ei Ei Hlaing, Narissara Lailerd, Sittiruk Roytrakul, Pichapat Piamrojanaphat

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Type 2 diabetes is one of the most common metabolic diseases all over the world. The pathogenesis of type 2 diabetes is not the only dysfunction of pancreatic beta cells but also insulin resistance in muscle, liver and adipose tissue. High levels of circulating free fatty acids, an increased lipid content of muscle cells, impaired insulin-mediated glucose uptake and diminished mitochondrial functioning are pathophysiological hallmarks of diabetic skeletal muscles. Purple rice (Oryza sativa L. indica) has been shown to have antidiabetic effects. However, the underlying mechanism(s) of antidiabetic activity of purple rice is still unraveled. In this research, to explore in-depth cellular mechanism(s), proteomic profile of purple rice supplemented type 2 diabetic rats’ skeletal muscle were analyzed contract with non-supplemented rats. Diabetic rats were induced high-fat diet combined with streptozotocin injection. By using one- dimensional gel electrophoresis (1-DE) and LC-MS/MS quantitative proteomic method, we analyzed proteomic profiles in skeletal muscle of normal rats, normal rats with purple rice supplementation, type 2 diabetic rats, and type 2 diabetic rats with purple rice supplementation. Total 2676 polypeptide expressions were identified. Among them, 24 peptides were only expressed in type 2 diabetic rats, and 24 peptides were unique peptides in type 2 diabetic rats with purple rice supplementation. Acetyl CoA carboxylase 1 (ACACA) found as unique protein in type 2 diabetic rats which is the major enzyme in lipid synthesis and metabolism. Interestingly, DNA damage response protein, heterogeneous nuclear ribonucleoprotein K [Mus musculus] (Hnrnpk), was upregulated in type 2 diabetic rats’ skeletal muscle. Meanwhile, unique proteins of type 2 diabetic rats with purple rice supplementation (bone morphogenetic 7 protein preproprotein, BMP7; and forkhead box protein NX4, Foxn4) involved with muscle cells growth through the regulation of TGF-β/Smad signaling network. Moreover, BMP7 may effect on insulin signaling through the downstream signaling of protein kinase B (Akt) which acts in protein synthesis, glucose uptake, and glycogen synthesis. In conclusion, our study supports that type 2 diabetes impairs muscular lipid metabolism. In addition, purple rice might recover the muscle cells growth and insulin signaling.

Keywords: proteomics, purple rice bran, skeletal muscle, type 2 diabetic rats

Procedia PDF Downloads 244

Authors: Liat Kulik

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The main goal of the present study was to examine gender differences in the variables that explain the experience of role conflict and well-being among Jewish working fathers and mothers in the Israel. The experience of work-family conflict arises from simultaneous pressures from the work and family domains that are mutually incompatible. In light of the expansion of women's role set following the addition of paid employment outside of the home, most of the studies dealing with the impact of multiple roles on well-being have been conducted among women. However, changes in gender roles in recent years have also affected men's role set, as reflected in the terms ‘new men’ and ‘new fathers’. Based on structural equation modeling, the study examined gender differences in variables that explain the experience of two types of role conflict – family interferes with work (FIW) and work interferes with family (WIF), as well as with the sense of well-being (positive and negative affect) among 611 employed Jewish mothers and fathers in Israel. The findings revealed that for women, both FIW and WIF conflict correlated negatively with well-being, whereas for men, a negative correlation with well-being was found only in the case of FIW conflict. For both men and women, egalitarian gender role ideology correlated with the dimension of positive effect, but the correlation was stronger for men. The findings highlight the contribution of egalitarian gender role ideology to alleviating the experience of role conflict and improving the emotional well-being of both men and women. Contrary to expectations, social support contributed more to mitigating negative effect among men than women. On the whole, the findings highlight the changes that men have experienced in the work-family system. In sum, the research findings shed new light on the masculine image in terms of the experience of FIW conflict. In contrast to the prevailing assumption that FIW role conflict is predominant among women, the findings of this study indicate that today, this type of role conflict is experienced equally by men and women whereas WIF conflict is predominant among men. Furthermore, contrary to expectations, levels of perceived social support were found to be similar for men and women, and men benefited from it even more than women did.

Keywords: FIW conflict, WIF conflict, social support, egalitarian gender role ideology, overload

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Authors: P. N. Okeke, J. N. Chikwendu

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The study evaluated the effects of varying fermentation periods on the nutrients and anti-nutrients composition of African yam bean (Sphenostylis stenocarpa) and acha (Digitaria exilis) flour blends and sensory properties of their products. The African yam bean seeds and acha grains were fermented for 24 hrs, 48 and 72 hrs, dried (sun drying) and milled into fine flour. The fermented flours were used in a ratio of 70:30 (Protein basis) to formulate composite flour for meat pie and biscuits production. Both the fermented and unfermented flours and products were analyzed for chemical composition using the standard method. The data were statistically analyzed using SPSS version 15 to determine the mean and standard deviation. The 24, 48, and 72 hrs fermentation periods increased protein (22.81, 26.15 and 24.00% respectively). The carbohydrate, ash and moisture contents of the flours were also increased as a result of fermentation (68.01-76.83, 2.26-4.88, and 8.36-13.00% respectively). The 48 hrs fermented flour blends had the highest increase in ash relative to the control (4.88%). Fermentation increased zinc, iron, magnesium and phosphorus content of the flours. Treatment drastically reduced the anti-nutrient (oxalate, saponin, tannin, phytate, and hemagglutinin) levels of the flours. Both meat pie and biscuits had increased protein relative to the control (27.36-34.28% and 23.66-25.09%). However, the protein content of the meat pie increased more than that of the biscuits. Zinc, Iron, Magnesium and phosphorus levels increased in both meat pie and biscuits. Organoleptic attributes of the products (meat pie and biscuits) were slightly lower than the control except those of the 72 hrs fermented flours.

Keywords: fermentation, African yam bean, acha, biscuits, meat-pie

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Authors: Özlem Ateş Sönmezoğlu, Begüm Terzi, Ahmet Yıldırım, Leyla Gündüz

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Durum wheat quality is defined as its suitability for pasta processing, that is pasta making quality. Another factor that determines the quality of durum wheat is the nutritional value of wheat or its final products. Wheat is a basic source of calories, proteins and minerals for humans in many countries of the world. For this reason, improvement of wheat nutritional value is of great importance. In recent years, deficiencies in protein and micronutrients, particularly in iron and zinc, have seriously increased. Therefore, basic foods such as wheat must be improved for micronutrient content. The effects of some major genes for grain quality established. Gpc-B1 locus is one of the genes increased protein and micronutrients content, and used in improvement studies of durum wheat nutritional value. The aim of this study was to increase the protein content and the micronutrient (Fe, Zn ve Mn) contents of an advanced durum wheat line (TMB 1) that was previously improved for its protein quality. For this purpose, TMB1 advanced durum wheat line were used as the recurrent parent and also, UC1113-Gpc-B1 line containing the Gpc-B1 gene was used as the gene source. In all of the generations, backcrossed plants carrying the targeted gene region were selected by marker assisted selection (MAS). BC4F1 plants MAS method was employed in combination with embryo culture and rapid plant growth in a controlled greenhouse conditions in order to shorten the duration of the transition between generations in backcross breeding. The Gpc-B1 gene was selected specific molecular markers. Since Yr-36 gene associated with Gpc-B1 allele, it was also transferred to the Gpc-B1 transferred lines. Thus, the backcrossed plants selected by MAS are resistance to yellow rust disease. This research has been financially supported by TÜBİTAK (112T910).

Keywords: Durum wheat, Gpc-B1, MAS, Triticum durum, Yr-36

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Authors: Shabnam Abdi, Behzad Toosi

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Background: Melanoma is the most lethal type of malignant skin cancer in humans and dogs since it spreads rapidly throughout the body. Despite significant advances in treatment, cancer at an advanced stage has a poor prognosis. Hence, more effective treatments are needed to enhance outcomes with fewer side effects. Erythropoietin-producing hepatocellular receptors are the largest family of receptor tyrosine kinases and are divided into two subfamilies, EphA and EphB, both of which play a significant role in disease, especially cancer. Due to their association with proliferation and invasion in many aggressive types of cancer, Eph receptor tyrosine kinases (Eph RTKs) are promising cancer therapy molecules. Because these receptors have not been studied in canine melanoma, we investigated how EphA2 influences survival and tumorigenicity of melanoma cells. Methods: Expression of EphA2 protein in canine melanoma cell lines and human melanoma cell line was evaluated by Western blot. Melanoma cells were transduced with lentiviral particles encoding Eph-targeting shRNAs or non-silencing shRNAs (control) for silencing the expression of EphA2 receptor, and silencing was confirmed by Western blotting and immunofluorescence. The effect of siRNA treatment on cellular proliferation, colony formation, tumorsphere assay, invasion was analyzed by Resazurin assay Matrigel invasion assay, respectively. Results: Expression of EphA2 was detected in canine and human melanoma cell lines. Moreover, stably silencing EphA2 by specific shRNAs significantly and consistently decreased the expression of EphA2 protein in both human and canine melanoma cells. Proliferation, colony formation, tumorsphere and invasion of melanoma cells were significantly decreased in EphA2 siRNA-treated cells compared to control. Conclusion: Our data provide the first functional evidence that the EphA2 receptor plays a critical role in the malignant cellular behavior of melanoma in both human and dogs.

Keywords: ephA2, targeting, melanoma, human, canine

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Authors: Orkide Donma, Mustafa M. Donma

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Obesity is associated with increased fat mass as well as fat percentage. Minerals are the elements, which are of vital importance. In this study, the relationships between body as well as bone mineral profile and the percentage as well as mass values of fat, fat-free portion, protein, skeletal muscle were evaluated in adult men with normal body mass index (N-BMI), and those classified according to different stages of obesity. A total of 103 adult men classified into five groups participated in this study. Ages were within 19-79 years range. Groups were N-BMI (Group 1), overweight (OW) (Group 2), first level of obesity (FLO) (Group 3), second level of obesity (SLO) (Group 4) and third level of obesity (TLO) (Group 5). Anthropometric measurements were performed. BMI values were calculated. Obesity degree, total body fat mass, fat percentage, basal metabolic rate (BMR), visceral adiposity, body mineral mass, body mineral percentage, bone mineral mass, bone mineral percentage, fat-free mass, fat-free percentage, protein mass, protein percentage, skeletal muscle mass and skeletal muscle percentage were determined by TANITA body composition monitor using bioelectrical impedance analysis technology. Statistical package (SPSS) for Windows Version 16.0 was used for statistical evaluations. The values below 0.05 were accepted as statistically significant. All the groups were matched based upon age (p > 0.05). BMI values were calculated as 22.6 ± 1.7 kg/m², 27.1 ± 1.4 kg/m², 32.0 ± 1.2 kg/m², 37.2 ± 1.8 kg/m², and 47.1 ± 6.1 kg/m² for groups 1, 2, 3, 4, and 5, respectively. Visceral adiposity and BMR values were also within an increasing trend. Percentage values of mineral, protein, fat-free portion and skeletal muscle masses were decreasing going from normal to TLO. Upon evaluation of the percentages of protein, fat-free portion and skeletal muscle, statistically significant differences were noted between NW and OW as well as OW and FLO (p < 0.05). However, such differences were not observed for body and bone mineral percentages. Correlation existed between visceral adiposity and BMI was stronger than that detected between visceral adiposity and obesity degree. Correlation between visceral adiposity and BMR was significant at the 0.05 level. Visceral adiposity was not correlated with body mineral mass but correlated with bone mineral mass whereas significant negative correlations were observed with percentages of these parameters (p < 0.001). BMR was not correlated with body mineral percentage whereas a negative correlation was found between BMR and bone mineral percentage (p < 0.01). It is interesting to note that mineral percentages of both body as well as bone are highly affected by the visceral adiposity. Bone mineral percentage was also associated with BMR. From these findings, it is plausible to state that minerals are highly associated with the critical stages of obesity as prominent parameters.

Keywords: bone, men, minerals, obesity

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

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

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

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