Search results for: cap-dependent protein translation

Authors: L. K. Davis

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The effects of the evolution force are observable in nature at all structural levels ranging from small molecular systems to conversely enormous biospheric systems. However, the evolution force and work associated with formation of biological structures has yet to be described mathematically or theoretically. In addressing the conundrum, we consider evolution from a unique perspective and in doing so we introduce the “Fundamental Theory of the Evolution Force: FTEF”. We utilized synthetic evolution artificial intelligence (SYN-AI) to identify genomic building blocks and to engineer 14-3-3 ζ docking proteins by transforming gene sequences into time-based DNA codes derived from protein hierarchical structural levels. The aforementioned served as templates for random DNA hybridizations and genetic assembly. The application of hierarchical DNA codes allowed us to fast forward evolution, while dampening the effect of point mutations. Natural selection was performed at each hierarchical structural level and mutations screened using Blosum 80 mutation frequency-based algorithms. Notably, SYN-AI engineered a set of three architecturally conserved docking proteins that retained motion and vibrational dynamics of native Bos taurus 14-3-3 ζ.

Keywords: 14-3-3 docking genes, synthetic protein design, time-based DNA codes, writing DNA code from scratch

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Authors: Mihai Palade, Gina Cecilia Pistol, Mariana Stancu, Veronica Chedea, Ionelia Taranu

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Nutrition is an important determinant of general health status, with especially focus on prevention and/or attenuation of the inflammatory-associated pathologies. People with chronic kidney disease can experience chronic inflammation that can lead to cardiovascular disease and even an increased rate of death. There are important links between chronic kidney diseases, inflammation and nutritional strategies that may prevent or protect against undesirable inflammation and oxidative stress. The grape by-products either seeds or pomace are rich in polyphenols which may be beneficial in prevention of inflammatory, antioxidant and antimicrobial processes. As a model for studying the impact of grape seeds on renal inflammation and oxidative stress, we used in this study weaned piglets. After a feeding trial of 30 days with a control diet and an experimental diet containing 5% grape seed (GS), kidney samples were collected. In renal tissues were determined the expression and activity of important markers of immune respose and oxidative stress: pro-inflammatory cytokines (TNF-alpha, IL-1 beta, IL-6, IL-8, IFN-gamma), anti-inflammatory cytokines (IL-4, IL-10), anti-oxidant enzymes (catalase CAT, superoxide dismutase SOD, glutathione peroxidise GPx) and important mediators belonging to nuclear receptors (NF-kB1, Nrf-2 and PPAR-gamma). Gene expression was evaluated by qPCR, whereas protein concentration was determined using proteomic techniques (ELISA). The activity of anti-oxidant enzymes was determined using specific kits. Our results showed that GS enriched in polyphenols does not have effect on TNF-alpha, IL-6 and IL-1 beta gene expression and protein concentration in kidney. By contrast, the gene expression and protein level of IL-8 and IFN-gamma were decreased in GS kidney. Anti-inflammatory cytokines IL-4 and IL-10 gene levels were increased in kidneys collected from GS piglets in comparison with controls, with no modification of protein levels between the two groups. The activities of anti-oxidant enzymes CAT and GPx were increased in kidney by GS, whereas SOD activity was unmodified in comparison with control samples. Also, the GS diet was associated with no modulation of mRNAs for nuclear receptors NF-kB1, Nrf-2 and PPAR-gamma gene expressions in kidneys. In conclusion, our results demonstrated that GS enriched in bioactive compounds such polyphenols could modulate inflammation and oxidative stress markers in kidney tissues. Further studies are necessary to elucidate the mechanism of action of GS compounds in case kidney inflammation associated with oxidative stress, and signalling molecules involved in these mechanisms.

Keywords: animal model, kidney inflammation, oxidative stress, grape seed

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Authors: Adrian Sebastian, Chris Gillespie

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We describe a case of diffuse rectal involvement with cap polyposis, manifesting with a protein-losing colopathy and occurring in the setting of advanced mechanical pelvic floor dysfunction. A 59-year-old male with a 5-year history of persistent excessive flatulence, defecatory difficulties, and diarrhea. He had extensive cap polyposis of the entire rectum endoscopically. His symptoms progressed to severe fecal incontinence with mucus leakage, pelvic pain, weight loss, and hypoalbuminemia. Clinical examination exhibited severe perineal descent, a large rectocele, poor anal squeeze, and a poor defecatory technique. After a trial of nonoperative therapies addressing his defecatory dysfunction, and Helicobacter pylori eradication, surgical resection was offered due to severe symptoms with ongoing incontinence and protein loss with no other reasonable options. A robotic abdominoperineal resection with a permanent colostomy was performed, followed by an uncomplicated recovery. Our observation of coexisting mechanical pelvic floor changes in this patient lends weight to the concept of a prolapse-related phenomenon in the pathophysiology of this rare condition.

Keywords: cap polyposis, pelvic dysfunction, fecal incontinence, case report

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Authors: Ramalingam Boobalan, Chinpiao Chen, Jui-I. Chiao

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A series of erlotinib analogues that have structural modification at 6,7-alkoxyl positions is efficiently synthesized. The key reactions that involved in synthesis are one-pot oxime formation-dehydration for the formation of nitrile, quinazoline ring formation reaction between aniline and o-cyanoaniline via formamidine intermediate, Fe/NH4Cl catalyzed reduction-hetereocyclization-reductive ring opening reaction for the formation of o-aminobenzamide, high yielding seal tube reactions for O-demethylation, sodium iodide substitution, ammonia substitution. The in vitro anti-tumor activity of synthesized compounds is studied in two non-small cell lung cancer (NSCLC) cell lines (A549 and H1975). Among the synthesized compounds, the iodo compound 6 (ETN-6) exhibits higher anti-cancer activity compared to erlotinib. An efficient method is developed for the conjugation of erlotinib analogue-4, alcohol compound, with protein, bovine serum albumin (BSA), via succinic acid linker. The in vitro anti-tumor activity of the protein attached erlotinib analogue, 8 (ETN-4-Suc-BSA), showed stronger inhibitory activity in both A549 and H1975 NSCLC cell lines.

Keywords: anti-cancer, BSA, EGFR, Erlotinib

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Authors: Kevin Kellner, Nga Lao, Orla Coleman, Paula Meleady, Niall Barron

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Chinese Hamster Ovary (CHO) cells are the prominent cell line used in biopharmaceutical production. To improve yields and find beneficial bioprocess phenotypes genetic engineering plays an essential role in recent research. The miR-23 cluster, specifically miR-24 and miR-27, was first identified as differentially expressed during hypothermic conditions suggesting a role in proliferation and productivity in CHO cells. In this study, we used sponge decoy technology to stably deplete the miRNA expression of the cluster. Furthermore, we implemented the CRISPR/Cas9 system to knockdown miRNA expression. Sponge constructs were designed for an imperfect binding of the miRNA target, protecting from RISC mediated cleavage. GuideRNAs for the CRISPR/Cas9 system were designed to target the seed region of the miRNA. The expression of mature miRNA and precursor were confirmed using RT-qPCR. For both approaches stable expressing mixed populations were generated and characterised in batch cultures. It was shown, that CRISPR/Cas9 can be implemented in CHO cells with achieving high knockdown efficacy of every single member of the cluster. Targeting of one miRNA member showed that its genomic paralog is successfully targeted as well. The stable depletion of miR-24 using CRISPR/Cas9 showed increased growth and specific productivity in a CHO-K1 mAb expressing cell line. This phenotype was further characterized using quantitative label-free LC-MS/MS showing 186 proteins differently expressed with 19 involved in proliferation and 26 involved in protein folding/translation. Targeting miR-27 in the same cell line showed increased viability in late stages of the culture compared to the control. To evaluate the phenotype in an industry relevant cell line; the miR-23 cluster, miR-24 and miR-27 were stably depleted in a Fc fusion CHO-S cell line which showed increased batch titers up to 1.5-fold. In this work, we highlighted that the stable depletion of the miR-23 cluster and its members can improve the bioprocess phenotype concerning growth and productivity in two different cell lines. Furthermore, we showed that using CRISPR/Cas9 is comparable to the traditional sponge decoy technology.

Keywords: Chinese Hamster ovary cells, CRISPR/Cas9, microRNAs, sponge decoy technology

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Authors: Reem Al-Shidhani, Isabelle S. R. Storer, Michael J. Bromley, Lydia Tabernero

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Invasive fungal diseases are an increasing global health concern that contributes to the high mortality rates in immunocompromised patients. The rising of antifungal resistance severely lowers the efficacy of the limited antifungal agents available. New antifungal drugs that target new mechanisms are necessary to tackle the current shortfalls. Amongst post- modifications, phosphorylation is a predominant and an outstanding protein alteration in all eukaryotes. In fungi, protein phosphorylation plays a vital role in many signal transduction pathways, including cell cycle, cell growth, metabolism, transcription, differentiation, proliferation, and virulence. The investigation of Aspergillus fumigatus phosphatases revealed seven genes essential for viability. Inhibiting one of these phosphatases is a new interesting route to develop novel antifungal drugs. In this study, we carried out an early drug discovery process targeting oneessential phosphatase, GlcA. Here, we report the identification of new GlcA inhibitors that show antifungal activity. These important finding open a new avenue to the development of novel antifungals to expand the current narrow arsenal of clinical candidates.

Keywords: invasive fungal diseases, phosphatases, GlcA, competitive inhibitors

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Authors: Adriana Muñoz-Talavera, Ana Rosa Rincón-Sánchez, Abraham Escobedo-Moratilla, María Cristina Islas-Carbajal, Miguel Ángel Gómez-Lim

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The highest rates of diabetes incidence and prevalence worldwide will increase the number of diabetic patients requiring insulin or insulin analogues. Then, current production systems would not be sufficient to meet the future market demands. Therefore, developing efficient expression systems for insulin and insulin analogues are needed. In addition, insulin analogues with better pharmacokinetics and pharmacodynamics properties and without mitogenic potential will be required. SCI-57 (single chain insulin-57) is an insulin analogue having 10 times greater affinity to the insulin receptor, higher resistance to thermal degradation than insulin, native mitogenicity and biological effect. Plants as expression platforms have been used to produce recombinant proteins because of their advantages such as cost-effectiveness, posttranslational modifications, absence of human pathogens and high quality. Immunoglobulin production with a yield of 50% has been achieved by transient expression in Nicotiana benthamiana (Nb). The aim of this study is to produce SCI-57 by transient expression in Nb. Methodology: DNA sequence encoding SCI-57 was cloned in pICH31070. This construction was introduced into Agrobacterium tumefaciens by electroporation. The resulting strain was used to infiltrate leaves of Nb. In order to isolate SCI-57, leaves from transformed plants were incubated 3 hours with the extraction buffer therefore filtrated to remove solid material. The resultant protein solution was subjected to anion exchange chromatography on an FPLC system and ultrafiltration to purify SCI-57. Detection of SCI-57 was made by electrophoresis pattern (SDS-PAGE). Protein band was digested with trypsin and the peptides were analyzed by Liquid chromatography tandem-mass spectrometry (LC-MS/MS). A purified protein sample (20µM) was analyzed by ESI-Q-TOF-MS to obtain the ionization pattern and the exact molecular weight determination. Chromatography pattern and impurities detection were performed using RP-HPLC using recombinant insulin as standard. The identity of the SCI-57 was confirmed by anti-insulin ELISA. The total soluble protein concentration was quantified by Bradford assay. Results: The expression cassette was verified by restriction mapping (5393 bp fragment). The SDS-PAGE of crude leaf extract (CLE) of transformed plants, revealed a protein of about 6.4 kDa, non-present in CLE of untransformed plants. The LC-MS/MS results displayed one peptide with a high score that matches SCI-57 amino acid sequence in the sample, confirming the identity of SCI-57. From the purified SCI-57 sample (PSCI-57) the most intense charge state was 1069 m/z (+6) on the displayed ionization pattern corresponding to the molecular weight of SCI-57 (6412.6554 Da). The RP-HPLC of the PSCI-57 shows the presence of a peak with similar retention time (rt) and UV spectroscopic profile to the insulin standard (SCI-57 rt=12.96 and insulin rt=12.70 min). The collected SCI-57 peak had ELISA signal. The total protein amount in CLE from transformed plants was higher compared to untransformed plants. Conclusions: Our results suggest the feasibility to produce insulin analogue SCI-57 by transient expression in Nicotiana benthamiana. Further work is being undertaken to evaluate the biological activity by glucose uptake by insulin-sensitive and insulin-resistant murine and human cultured adipocytes.

Keywords: insulin analogue, mass spectrometry, Nicotiana benthamiana, transient expression

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Authors: Abdelaziz Messis, Azzeddine Bettache, Nawel Boucherba, Said Benallaoua, Mouloud Kecha

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Actinobacteria are of special biotechnological interest since they are known to produce chemically diverse compounds with a wide range of biological activity. This distinct clade of Gram-positve bacteria include some of the key antibiotic producers and are also sources of several bioactive compounds, established commercially a newly filamentous bacteria was recovered from Tikjda forest soil (Algeria) for its high antifungal activity against various pathogenic and phytopathogenic fungi. The nucleotide sequence of the 16S rRNA gene (1454 pb) of Streptomyces sp. TKJ2 exhibited close similarity (99 %) with other Streptomyces16S rRNA genes. Antifungal metabolite production of Streptomyces sp TKJ2 was evaluated using six different fermentation media. The extracellular products contained potent antifungal agents. Antifungal protein produced by Streptomyces sp. TKJ2 on PCA medium has been purified by ammonium sulfate precipitation, SPE column chromatography and high-performance liquid chromatography in a reverse-phase column. The UV chromatograms of the active fractions obtained at 214 nm by NanoLC-ESI-MS/MS have different molecular weights. The F20 Peptidic fraction obtained from culture filtrat of Streptomyces sp. TKJ2 precipitated at 30% of ammonium sulfate was selected for analysis by infusion ESI-MS which yielded a singly charged ion mass of 437.17 Da.

Keywords: actinobacteria, antifungal protein, chromatography, Streptomyces

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Authors: Leyla Sattarzadeh, Maghsoud Peeri, Mohammadali Azarbaijani, Hasan Matin Homaee

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Inflammation by various mechanisms may cause atherosclerosis. Systemic circulating inflammatory markers such as C-reactive protein (CRP), pro-inflammatory cytokines such as Interleukin-6 (IL-6) and adhesion molecules like Intracellular Adhesion Molecule-1 (ICAM-1) are the predictors of cardiovascular diseases. Regarding the conflicting results about the effect of resistance exercise training on these inflammatory markers, the present study aimed to examine the effect of eight week different patterns of resistance exercise training on CRP, IL-6 and ICAM-1 levels in healthy untrained women. 40 volunteered and healthy untrained female university students (aged: 21+ 3 yr., Body Mass Index: 21.5+ 3.5 kg/m2) were selected purposefully and divided into three groups. At the end of training protocol and after subjects drop during the protocol in upper body exercise training (n=11), lower body (n=12) completed the eight week of training period although the control group (n=7) did anything. Blood samples gathered pre and post experimental period and CRP, IL-6 and ICAM-1 levels were evaluated using special laboratory kits, then the difference of pre and post values of each indices analyzed using one way Analysis of Variance (α < 0.05). The results of one way ANOVA for difference of pre and post values of CRP and ICAM-1 showed no significant changes due to the exercise training. But there were significant differences between groups about IL-6. Tukey post- hoc test indicated that there is significant difference between the differences of pre and post values of IL-6 between lower body exercise training group and control group, and eight weeks of lower body exercise training lead to significant changes in IL-6 values. There were no changes in anthropometric indices. The findings show that the different patterns of upper and lower body exercise training by involving the different amount of muscles altered the IL-6 values in lower body exercise training group probably because of engaging the bigger amount of muscles, but showed any significant changes about CRP and ICAM-1 probably due to intensity and duration of exercise or the lower levels of these markers at baseline of healthy people.

Keywords: C-reactive protein, interleukin-6, intracellular adhesion molecule-1, resistance training

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Authors: Hugo F. Lobaton

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The cyanobacterium Spirulina platensis is a prokaryotic photoautotrophic microorganism that is successfully cultivated for the commercialization as whole biomass due to its high protein content and promising valuable substance. For instance, phycocyanin has recently drawn the interest of the food and cosmetic industries due to its bright blue colour and its strong antioxidant capacities. The phycocyanin (PC) is the main protein-pigment in S. platensis (4% to 20%). In batches, the rate of overproduction of metabolites by cyanobacteria is limited or activated by the depletion of required substrates. The aim of this study was to develop a kinetic law that describes phycocyanin formation during batch cultivation. S. platensis was cultivated in 1 L bubble column photobioreactor with 30°C and 700 µmol m⁻² s⁻¹. Culture samples were daily collected from the bubble columns in sterile conditions. The biomass (g l⁻¹) was measured directly after a biomass lyophilisation process, and phycocyanin extractions and measurements were done according to a well-established protocol. A kinetic law for phycocyanin formation that includes nitrate and bicarbonate limitations was proposed and linked to the biomass core model. The set of differential equations were solved in MATLAB. Concerning to product formation, the experimental results show that phycocyanin mass fraction is degraded as results of the complete nitrate depletion and nitrate additions during the cultivation help to keep constant this molecule until new macro-element limitation appear. According to the model, bicarbonate is this limitation.

Keywords: phycocyanin, nitrate, bicarbonate, spirulina

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Authors: Tzong-Ying Hao, Mohammad T. Rahmani

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This article presents the structural health monitoring (SHM) method based on changes in wave traveling times (wave method) within a layered 1-D shear beam model of structure. The wave method measures the velocity of shear wave propagating in a building from the impulse response functions (IRF) obtained from recorded data at different locations inside the building. If structural damage occurs in a structure, the velocity of wave propagation through it changes. The wave method analysis is performed on the responses of Torre Central building, a 9-story shear wall structure located in Santiago, Chile. Because events of different intensity (ambient vibrations, weak and strong earthquake motions) have been recorded at this building, therefore it can serve as a full-scale benchmark to validate the structural health monitoring method utilized. The analysis of inter-story drifts and the Fourier spectra for the EW and NS motions during 2010 Chile earthquake are presented. The results for the NS motions suggest the coupling of translation and torsion responses. The system frequencies (estimated from the relative displacement response of the 8th-floor with respect to the basement from recorded data) were detected initially decreasing approximately 24% in the EW motion. Near the end of shaking, an increase of about 17% was detected. These analysis and results serve as baseline indicators of the occurrence of structural damage. The detected changes in wave velocities of the shear beam model are consistent with the observed damage. However, the 1-D shear beam model is not sufficient to simulate the coupling of translation and torsion responses in the NS motion. The wave method is proven for actual implementation in structural health monitoring systems based on carefully assessing the resolution and accuracy of the model for its effectiveness on post-earthquake damage detection in buildings.

Keywords: Chile earthquake, damage detection, earthquake response, impulse response function, shear beam model, shear wave velocity, structural health monitoring, torre central building, wave method

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Authors: Daruliza Kernain, Shaharum Shamsuddin, See Too Wei Cun

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CTCF is a unique, highly conserved and ubiquitously expressed 11 zinc finger (ZF) transcriptional factor with multiple target sites. It is able to bind to various target sequences to perform different regulatory roles including promoter activation or repression, creating hormone-responsive gene silencing element, and functional block of enhancer-promoter interactions. The binding of CTCF to the essential binding site is through the combination of different ZF domain. On the other hand, BORIS for brother of the regulator of imprinted sites, which expressed only in the testis and certain cancer cell line is homology to CTCF 11 ZF domains. Since both transcriptional factors share the same ZF domains hence there is a possibility for both to bind to the same target sequences. In this study, the interaction of these two proteins to multi-functional Y-box DNA/RNA-binding factor, YB-1 was determined. The protein-protein interaction between CTCF/YB-1 and BORIS/YB-1 were discovered by Co-immuno-precipitation (CO-IP) technique through reciprocal experiment from RGBM total cell lysate. The results showed that both CTCF and BORIS were able to interact with YB-1 in Glioma RGBM cell line. To the best of our knowledge, this is the first findings demonstrating the ability of BORIS and YB-1 to form a complex in vivo.

Keywords: immunoprecipitation, CTCF/BORIS/YB-1, transcription factor, molecular medicine

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Authors: Cenza Rhoda, Falone Sunda, Elvis Kidzeru, Nonhlanhla P. Khumalo, Afolake Arowolo

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Introduction: The human FAM111B gene mutations are associated with POIKTMP, a rare multi-organ fibrosing disease. Recent studies also reported the overexpression of FAM111B in specific cancers. However, the role of FAM111B in these pathologies, particularly fibrosarcoma, remains unknown. Materials and Methods: FAM111B RNA expression in some cancer cell lines was assessed in silico and validated in vitro in these cell lines and skin fibroblasts derived from the South African family member affected by POIKTMP with the heterozygous FAM111B gene mutation: NM_198947.4: c.1861T>G (p. Tyr621Asp or Y621D) by qPCR and western blot. The cellular function of FAM111B was also studied in HT1080 using various cell-based functional assays and a simple and cost-effective PCR-RFLP method for genotyping/screening FAM111B gene mutations described. Results: Expression studies showed upregulated FAM111B mRNA and protein in the cancer cells. High FAM111B expression with robust nuclear localization occurred in HT1080. Additionally, expression data and cell-based assays indicated that FAM111B led to the upregulation of cell migration and decreased cell apoptosis and cell proliferation modulation. FAM111B Y621D mutation showed similar effects on cell migration but minimal impact on cell apoptosis. FAM111B mRNA and protein expression were markedly downregulated (p ≤ 0.05) in the patient's skin-derived fibroblasts. Lastly, the PCR-RFLP method successfully genotyped FAM111B Y621D gene mutation. Discussion: FAM111B is a cancer-associated nuclear protein: Its modulation by mutations may enhance cell migration and proliferation and decrease apoptosis, as seen in cancers and POIKTMP/fibrosis, thus representing a viable therapeutic target in these disorders. Furthermore, the PCR-RFLP method could prove a valuable tool for FAM111B mutation validation or screening in resource-constrained laboratories.

Keywords: FAM111B, POIKTMP, cancer, fibrosis, PCR-RFLP

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Authors: Taylor La Carriere

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“What I had madly possessed was not she, but my own creation, another, fanciful Lolita perhaps, more real than Lolita; overlapping, encasing he and having no will, no consciousness indeed, no life of her own,” Vladimir Nabokov writes in his seminal work, Lolita. Throughout Nabokov’s novel, the eponymous character is rendered nonexistent through unreliable narrator Humbert Humbert’s impenetrable narrative, infused with lyrical rationalization. Instead, Lolita is “safely solipsised,” as Humbert muses, solidifying the potential for the erasure of Lolita’s agency and identity. In this literary work, Lolita’s voice is reduced to a nearly invisible presence, only seen through the eyes of her captor. However, in Stanley Kubrick’s film adaptation of Lolita (1962), the “nymphet,” as Nabokov coins, reemerges with a voice of her own, fueled by a lyric impulse, that displaces Humbert’s first-person narration. The lyric, as defined by Catherine Ing, is the voice of the invisible; it is also characterized by performance, the concentrated utterance of individual emotion, and the appearance of spontaneity. The novel’s lyricism is largely in the service of Humbert’s “seductive” voice, while the film reorients it more to Lolita’s subjectivity. Through a close analysis of Kubrick’s cinematic techniques, this paper examines the emergence and translation of Lolita’s voice in contrast with Humbert’s attempts to silence her in Nabokov’s Lolita, hypothesizing that Kubrick translates Lolita’s presence into a visual and aural voice with lyrical attributes, exemplified through the establishment of an altered power dynamic, Sue Lyon’s transformative performance as the titular character, Nelson Riddle and Bob Harris’ musical score, and the omission of Humbert’s first-person point-of-view. In doing so, the film reclaims Lolita’s agency by taking instances of Lolita’s voice in the novel as depicted in the last half of the work and expanding upon them in a way only cinematic depictions could allow. The results of this study suggest that Lolita’s voice in Kubrick’s adaptation functions without disrupting the lyricism present in Nabokov’s source text, materializing through the actions, expressions, and performance of Sue Lyon in the film. This voice, fueled by a lyric impulse of its own, refutes the silence bestowed upon the titular character and enables its ultimate reclamation upon the silver screen.

Keywords: cinema, adaptation, Lolita, lyric voice

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Authors: F. A. M. Safwan, H. N. N. Dilrukshi, P. U. S. Peiris

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Less availability of high quality green forages leads to low productivity of national dairy herd of Sri Lanka. Growing grass and fodder to suit the production system is an efficient and economical solution for this problem. CO-3 is placed in a higher category, especially on tillering capacity, green forage yield, regeneration capacity, leaf to stem ratio, high crude protein content, resistance to pests and diseases and free from adverse factors along with other fodder varieties grown within the country. An experiment was designed to determine the effect of organic sheep manure, inorganic fertilizers and liquid sheep manure on growth, yield and nutritive value of CO-3. The study was consisted with three treatments; sheep manure (T1), recommended inorganic fertilizers (T2) and liquid sheep manure (T3) which was prepared using bucket fermentation method and each treatment was consisted with three replicates and those were assigned randomly. First harvest was obtained after 40 days of plant establishment and number of leaves (NL), leaf area (LA), tillering capacity (TC), fresh weight (FW) and dry weight (DW) were recorded and second harvest was obtained after 30 days of first harvest and same set of data were recorded. SPSS 16 software was used for data analysis. For proximate analysis AOAC, 2000 standard methods were used. Results revealed that the plants treated with T1 recorded highest NL, LA, TC, FW and DW and were statistically significant at first and second harvest of CO-3 (p˂ 0.05) and it was found that T1 was statistically significant from T2 and T3. Although T3 was recorded higher than the T2 in almost all growth parameters; it was not statistically significant (p ˃0.05). In addition, the crude protein content was recorded highest in T1 with the value of 18.33±1.61 and was lowest in T2 with the value of 10.82±1.14 and was statistically significant (p˂ 0.05). Apart from this, other proximate composition crude fiber, crude fat, ash, moisture content and dry matter were not statistically significant between treatments (p ˃0.05). In accordance with the results, it was found that the organic fertilizer is the best fertilizer for CO-3 in terms of growth parameters and crude protein content.

Keywords: fertilizer, growth parameters, Hybrid Napier CO-3, proximate composition

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Authors: Kongngern K., Homwonk C., Theerakulpisut P., Roytrakul R.

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Rice cultivation is crucial globally, especially in Asia. Soil salinity poses a significant challenge for agricultural lands. Understanding the expression patterns of different rice varieties under salt stress can provide insights for developing more salt-tolerant cultivars. This study aims to compare the expression patterns of two rice varieties, Thai moderately salt-tolerant rice (Leaung Anan) and high salt-tolerant rice (Pokkali), in response to salt stress. By analyzing protein expression, the research seeks to identify key proteins associated with salt tolerance in rice. The expression patterns of the two rice varieties under salt stress were analyzed using 1D-SDS-PAGE, NanoLC-MS/MS, and MEV software. These methods enabled the researchers to assess the differential expression of proteins in the leaf sheaths of the rice plants. These results indicate that the study identified 18 proteins, exhibited significantly different expression patterns between the two rice cultivars under salt stress. Notably, certain proteins, such as Os05g0364500 and pr1-like protein, showed contrasting expression profiles in the two varieties. The up-regulated proteins, predominantly observed in the salt-tolerant rice, may contribute to the survival of rice plants under salt stress and may provide valuable insights for breeding programs aiming to enhance salt tolerancein rice cultivars.

Keywords: mass spectrometry, proteomics, rice leaf sheaths, salt stress

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Authors: Yun-An Chen, Hung-Jun Lin, Tai-Horng Young, Der-Zen Liu

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Decellularized brain extracellular matrix had been shown that it has the ability to influence on cell proliferation, differentiation and associated cell phenotype. However, this scaffold is thought to have poor mechanical properties and rapid degradation, it is hard for cell recellularization. In this study, we used decellularized brain extracellular matrix combined with chitosan, which is naturally occurring polysaccharide and non-cytotoxic polymer, forming a 3-D scaffold for neural stem/precursor cells (NSPCs) regeneration. HE staining and DAPI fluorescence staining confirmed decellularized process could effectively vanish the cellular components from the brain. GAGs and collagen I, collagen IV were be showed a great preservation by Alcain staining and immunofluorescence staining respectively. Decellularized brain extracellular matrix was well mixed in chitosan to form a 3-D scaffold (DB-C scaffold). The pore size was approximately 50±10 μm examined by SEM images. Alamar blue results demonstrated NSPCs had great proliferation ability in DB-C scaffold. NSPCs that were cultured in this complex scaffold differentiated into neurons and astrocytes, as reveled by NSPCs expression of microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP). In conclusion, DB-C scaffold may provide bioinformatics cues for NSPCs generation and aid for CNS injury functional recovery applications.

Keywords: brain, decellularization, chitosan, scaffold, neural stem/precursor cells

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Authors: Payel Ghosh, Rama Chandra Pradhan, Sabyasachi Mishra

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Jamun (Syzygium cuminii L.) is one of the important indigenous minor fruit with high medicinal value. The jamun cultivation is unorganized and there is huge loss of this fruit every year. The perishable nature of the fruit makes its postharvest management further difficult. Due to the strong cell wall structure of pectin-protein bonds and hard seeds, extraction of juice becomes difficult. Enzymatic treatment has been commercially used for improvement of juice quality with high yield. The objective of the study was to optimize the best treatment method for juice extraction. Enzymes (Pectinase and Tannase) from different stains had been used and for each enzyme, best result obtained by using response surface methodology. Optimization had been done on the basis of physicochemical property, nutritional property, sensory quality and cost estimation. According to quality aspect, cost analysis and sensory evaluation, the optimizing enzymatic treatment was obtained by Pectinase from Aspergillus aculeatus strain. The optimum condition for the treatment was 44 ^oC with 80 minute with a concentration of 0.05% (w/w). At these conditions, 75% of yield with turbidity of 32.21NTU, clarity of 74.39%T, polyphenol content of 115.31 mg GAE/g, protein content of 102.43 mg/g have been obtained with a significant difference in overall acceptability.

Keywords: enzymatic treatment, Jamun, optimization, physicochemical property, sensory analysis

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Authors: Mehmet Celik, Celal Erbas, Mehtap Baykal, Aygül Kucukgulmez, Mahmut Ali Gokce, Bilge Kaan Tekelioglu

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In this study, determination of differences in fresh meat yield and proximate compositions of different weight groups of sea bream and sea bass grown in cages in Izmir region of the Aegean Sea were aimed. For this purpose, the length and weight of five different weight groups of sea bass (I: 175.8±5.2, II: 227.3±10.2, III: 293.3±21.3, IV: 404±9.9, V: 508.7±46 g) and sea bream (I: 146.6±13.6, II: 239.8±21.7, III: 279.2±20.8, IV: 400.9±10.5, V: 546.8±0.8 g) were measured and the amount of edible and non-edible parts were determined. Besides this, protein, lipid, dry matter, ash, condition factor, HSI and VSI values were compared according to different weight groups for each species. According to the results of analysis, while the absolute meat yields of sea bream was between 69-294 g, it was between 71-252 g for the sea bass and the highest meat yields were found in fifth (V) weight groups of fish for both species. The relative meat yield (%) was determined in weight group II for sea bass and in the IV. group in sea bream with 51.9%. However, the amount of muscle tissue lipids in I. and V. weight groups of sea bream ranged between 3.6 to 11.9 % and ranged between 6.2 to 9.0 % for sea bass respectively. Protein, fillet and ash content increased in direct proportion to the weight. As a result, it can be speculated that when the meat yield and lipid rates were considered, IV. group in sea bream and II. group in sea bass are the most advantageous groups for the consumers. Acknowledgement: This work was supported by the Scientific Research Project Unit of the University of Cukurova, Turkey under grant no FBA-2015-3830.

Keywords: sea bream, sea bass, meat yield, proximate composition, different weight

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Authors: Negar Ghotbeddin

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In this study, the changes in survival, growth, body composition, hematological, biochemical and immunological parameters of oscar fish (Astronotus ocellatus) have been investigated with dietary spirulina powder supplementation. Total of 300 fish with an initial weight of 8.37 ± 0.36 was distributed to three treatments and one control (0%). The fish were fed 8 weeks with diets containing different concentrations of S. powder: (control (0%), 2.5%, 5%, and 10%). Then sampling was done, and different parameters were measured by standard methods. Growth performance such as weight gain (%), specific growth rate (SGR) and feed conversion ratio (FCR) significantly improved in fish fed with S. powder (p < 0.5). Crude protein significantly increased in the S. powder supplemented groups (p < 0.5). However, crude lipid decreased with the increasing of dietary S. powder levels. Total protein increased in fish fed with 10% S. powder. Triglycerides and cholesterol decreased with the increasing of dietary S. powder levels. Immunological parameters including C3 and C4 increased significantly with the increasing of dietary S. powder levels, and lysozyme was improved in 10% S. powder. Results of this study indicated that S. powder had positive effects on Oscar fish and the best values were observed at 10 % S. powder.

Keywords: spirulina powder, growth performance, body composition, hematology, immunity, Astronotus ocellatus

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Authors: Saima Saleem, Zubair Abbasi, Abdul Hameed, Mansoor Ahmed Khan, Navid Rashid Qureshi, Abid Azhar

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Oral Squamous Cell Carcinoma (OSCC) is the leading cause of death in the developing countries like Pakistan. This problem aggravates because of the excessive use of available chewing products. In spite of widespread information on their use and purported legislations against their use the Pakistani markets are classical examples of selling chewable carcinogenic mutagens. Reported studies indicated that these products are rich in reactive oxygen species (ROS) and polyphenols. TP53 gene is involved in the suppression of tumor. It has been reported that somatic mutations caused by TP53 gene are the foundation of the cancer. This study aims to find the loss of TP53 functions due to mutation/polymorphism caused by genomic alteration and interaction with tobacco and its related ingredients. Total 260 tissues and blood specimens were collected from OSCC patients and compared with age and sex matched controls. Mutations in exons 2-11 of TP53 were examined by PCR-SSCP. Samples showing mobility shift were directly sequenced. Two mutations were found in exon 4 at nucleotide position 108 and 215 and one in exon 7 at nucleotide position 719 of the coding sequences in patient’s tumor samples. These results show that substitution of proline with arginine at codon 72 and serine with threonine at codon 240 of p53 protein. These polymorphic changes, found in tumor samples of OSCC, could be involved in loss of heterozygocity and apoptotic activity in the binding domain of TP53. The model of the mutated TP53 gene elaborated a nonfunctional unfolded p53 protein, suggesting an important role of these mutations in p53 protein inactivation and malfunction. This nonfunctional 3D model also indicates that exogenous tobacco related carcinogens may act as DNA-damaging agents affecting the structure of DNA. The interpretations could be helpful in establishing the pathways responsible for tumor formation in OSCC patients.

Keywords: TP53 mutation/polymorphism, OSCC, PCR-SSCP, direct DNA sequencing, 3D structure

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Authors: Maryam Ramezankhani

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The present research tries to study the possibility of formalizing the sense relation of hyponymy. It applied mathematical tools and also uses mathematical logic concepts especially those from propositional logic. In order to do so, firstly, it goes over the definitions of hyponymy presented in linguistic dictionaries and semantic textbooks. Then, it introduces a formal translation of the sense relation of hyponymy. Lastly, it examines the efficiency of the suggested formula by some examples of natural language.

Keywords: sense relations, hyponymy, formalizing, words’ sense relation, formalizing sense relations

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Authors: U.N. Emiri, E. Moroyei

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Mycoflora associated with the seed rot disease of Mucuna sloanei and their effects on nutrient and phytochemical composition of the seeds were investigated. The fungal pathogens implicated in the seed rot disease were Rhizopus stolonifer, Aspergillus flavus, Aspergillus niger, and Fusarium oxysporum. The fungal isolates were aseptically inoculated into healthy M. Sloanei seeds and incubated for 7 days at room temperature of 25 ± 30c. The results of the proximate and mineral analysis in mg/100g of fungal infected and non-infected (control) seeds that were carried out revealed that there was an increase in Moisture and Carbohydrate content of the fungal infected seeds relative to the non-infected seeds (control). However, there was a decrease in Ash, Fibre, Lipid, and Protein content of the fungal infected seeds relative to the non-infected (control). It was observed that moisture had increased from 10.50 ± 0.16 in the non-infected seeds to 17.60 ± 0.20 in the infected samples and Carbohydrate content had also increased from 49.6 ± 0.25 in the non-infected to 52.50 ± 0.29 in the infected seeds. The following parameters decreased in the infected than in the non-infected seeds. They include Ash 2.60 ± 0.12, Crude fibre 1.9 ± 0.08, Lipid 6.50 ± 0.16, and Protein content 18.50 ± 0.06. Similarly, Calcium 2.50 ± 0.12, Phosphorus 1.80 + 0.12 and Potassium 1.80 + 0.09 increased in the infected than in the non-infected seed, while iron 0.20 ± 0.05, Sodium 0.02 ± 0.01 and Magnesium 0.06 ± 0.02 decreased in the infected seeds. All phytochemical contents analyzed increased in the infected seeds viz Tannim 0.50 ± 0.12, Oxalate 1.60 ± 0.05, Hydrogen cyanide 1.82 ± 0.06, and Saponin 2.50+0.28. However, the nutrient compositions and Phytochemical between the infected and non-infected seeds are not significantly different (p > 0.05).

Keywords: Mycoflora, mucuna sloanei, seeds, phytochemical, nutrient composition

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Authors: Pritika Ramharack, Mahmoud E. S. Soliman

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The re-emerging Zika virus is an arthropod-borne virus that has been described to have explosive potential as a worldwide pandemic. The initial transmission of the virus was through a mosquito vector, however, evolving modes of transmission has allowed the spread of the disease over continents. The virus already been linked to irreversible chronic central nervous system (CNS) conditions. The concerns of the scientific and clinical community are the consequences of Zika viral mutations, thus suggesting the urgent need for viral inhibitors. There have been large strides in vaccine development against the virus but there are still no FDA-approved drugs available. Rapid rational drug design and discovery research is fundamental in the production of potent inhibitors against the virus that will not just mask the virus, but destroy it completely. In silico drug design allows for this prompt screening of potential leads, thus decreasing the consumption of precious time and resources. This study demonstrates an optimized and proven screening technique in the discovery of two potential small molecule inhibitors of Zika virus Methyltransferase and RNA-dependent RNA polymerase. This in silico “per-residue energy decomposition pharmacophore” virtual screening approach will be critical in aiding scientists in the discovery of not only effective inhibitors of Zika viral targets, but also a wide range of anti-viral agents.

Keywords: NS5 protein inhibitors, per-residue decomposition, pharmacophore model, virtual screening, Zika virus

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Authors: Raquel P. F. Guiné, Marlene I. C. Tenreiro, Ana C. Correia, Paulo Barracosa, Paula M. R. Correia

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The objective of this study was to evaluate the physical and chemical characteristics of Serra da Estrela cheese and compare these results with those of the sensory analysis. For the study were taken six samples of Serra da Estrela cheese produced with 6 different ecotypes of thistle in a dairy situated in Penalva do Castelo. The chemical properties evaluated were moisture content, protein, fat, ash, chloride and pH; the physical properties studied were color and texture; and finally a sensory evaluation was undertaken. The results showed moisture varying in the range 40-48%, protein in the range 15-20%, fat between 41-45%, ash between 3.9-5.0% and chlorides varying from 1.2 to 3.0%. The pH varied from 4.8 to 5.4. The textural properties revealed that the crust hardness is relatively low (maximum 7.3 N), although greater than flesh firmness (maximum 1.7 N), and also that these cheeses are in fact soft paste type, with measurable stickiness and intense adhesiveness. The color analysis showed that the crust is relatively light (L* over 50), and with a predominant yellow coloration (b* around 20 or over) although with a slight greenish tone (a* negative). The results of the sensory analysis did not show great variability for most of the attributes measured, although some differences were found in attributes such as crust thickness, crust uniformity, and creamy flesh.

Keywords: chemical composition, color, sensorial analysis, Serra da Estrela cheese, texture

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Authors: Samane Eftekhari Ranjbar

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Diabetes mellitus, a metabolic disorder characterized by elevated blood glucose levels, ranks among the leading causes of adult mortality. This study investigates the impact of an eight-week high-intensity interval training (HIIT) program combined with sodium nitrite supplementation on TNF- α, MURF1, and PI3K in a type 2 diabetes rodent model. Elevated TNF-α levels have been associated with insulin resistance, while MURF1 and PI3K play roles in muscle atrophy and insulin signaling pathways, respectively. In this experimental study, 15 eight-week-old rats from the Sara Laboratory Center in Tabriz were assigned to one of five groups: healthy control, diabetic control, diabetic with sodium nitrite supplementation, diabetic with eight weeks of intermittent exercise, and diabetic with eight weeks of interval training plus sodium nitrite supplementation. The HIIT protocol was designed to span eight weeks, with five weekly sessions at specified intensities and durations. Sodium nitrite, known for its vasodilatory and cytoprotective properties, was administered via injection. The findings revealed that the HIIT program and sodium nitrite supplementation influenced the examined biomarkers. ANOVA test outcomes indicated statistically significant differences in TNF- α (P=0.001), MURF1 (P=0.001), and PI3K (P=0.001) concentrations among the various groups. The healthy control group exhibited substantially decreased TNF- α, and MURF1 levels, as well as elevated PI3K levels compared to the diabetic control group. The exercise group, in conjunction with sodium nitrite supplementation, demonstrated a significant rise in PI3K levels (P=0.001) and a decline in TNF- α levels (P=0.018) relative to the diabetic control group. These results suggest that the combined intervention may help improve insulin sensitivity and reduce inflammation. However, MURF1 levels, which are related to muscle atrophy, showed no significant difference (P=0.24). In conclusion, in type 2 diabetic rats, an eight-week high-intensity interval training program with sodium nitrite supplementation does not affect MURF1 levels but does influence PI3K and TNF- α levels. This combination may hold potential for improving insulin sensitivity and reducing inflammation in type 2 diabetes patients, warranting further investigation and potential translation to human clinical trials.

Keywords: high-intensity interval training, sodium nitrate supplementation, type 2 diabetes, tumor necrosis factor-alpha, phosphatidylinositol-3-kinase, muscle RING-finger protein-1

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Authors: Kyle Phillips, Ndiko Ludidi

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Global climate change has resulted in altered rainfall patterns, which has resulted in annual losses in maize crop yields due to drought. Therefore it is important to produce maize cultivars that are more drought-tolerant, which is not an easily accomplished task as plants have a plethora of physical and biochemical adaptation methods. One such mechanism is the drought-induced expression of enzymatic and non-enzymatic proteins which assist plants to resist the effects of drought on their growth and development. One of these proteins is AtRD22 which has been identified in Arabidopsis thaliana. Using an in silico approach, a maize protein with 48% sequence homology to AtRD22 has been identified. This protein appears to be localized in the extracellular matrix, similarly to AtRD22. Promoter analysis of the encoding gene reveals cis-acting elements suggestive of induction of the gene’s expression by abscisic acid (ABA). Semi-quantitative transcriptomic analysis of the putative maize RD22 has revealed an increase in transcript levels after the exposure to drought. Current work elucidates the effect of up-regulation and silencing of the maize RD22 gene on the tolerance of maize to drought. The potential role of the maize RD22 gene in maize drought tolerance can be used as a tool to improve food security.

Keywords: abscisic acid, drought-responsive cis-acting elements, maize drought tolerance, RD22

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Authors: Gulmira Zhakupova, Tamara Tultabayeva, Aknur Muldasheva, Assem Sagandyk

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The development of technology and production of products with increased biological value based on the use of natural food raw materials are important tasks in the policy of the food market of the Republic of Kazakhstan. For Kazakhstan, livestock farming, in particular sheep farming, is the most ancient and developed industry and way of life. The history of the Kazakh people is largely connected with this type of agricultural production, with established traditions using dairy products from sheep's milk. Therefore, the development of new technologies from sheep’s milk remains relevant. In addition, one of the most promising areas for the development of food technology for therapeutic and prophylactic purposes is sheep milk products as a source of protein, immunoglobulins, minerals, vitamins, and other biologically active compounds. This article presents the results of research on the study of milk processing technology. The objective of the study is to study the possibilities of processing sheep milk and its role in human nutrition, as well as the results of research to improve the technology of sheep milk products. The studies were carried out on the basis of sanitary and hygienic requirements for dairy products in accordance with the following test methods. To perform microbiological analysis, we used the method for identifying Salmonella bacteria (Horizontal method for identifying, counting, and serotyping Salmonella) in a certain mass or volume of product. Nutritional value is a complex of properties of food products that meet human physiological needs for energy and basic nutrients. The protein mass fraction was determined by the Kjeldahl method. This method is based on the mineralization of a milk sample with concentrated sulfuric acid in the presence of an oxidizing agent, an inert salt - potassium sulfate, and a catalyst - copper sulfate. In this case, the amino groups of the protein are converted into ammonium sulfate dissolved in sulfuric acid. The vitamin composition was determined by HPLC. To determine the content of mineral substances in the studied samples, the method of atomic absorption spectrophotometry was used. The study identified the technological parameters of sheep milk products and determined the prospects for researching sheep milk products. Microbiological studies were used to determine the safety of the study product. According to the results of the microbiological analysis, no deviations from the norm were identified. This means high safety of the products under study. In terms of nutritional value, the resulting products are high in protein. Data on the positive content of amino acids were also obtained. The results obtained will be used in the food industry and will serve as recommendations for manufacturers.

Keywords: dairy, milk processing, nutrition, colostrum

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Authors: Saeed Ahmed, Muhammad Imran, Yasir Allah Ditta, Shahid Mehmood, Zahid Rasool

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A study was conducted to investigate the effect of different levels of microencapsulated butyric (MEB) on growth performance, gut health and immunity in commercial broiler chickens. In total, 336 day-old Hubbard classic broilers chicks were randomly assigned to 4 dietary treatments (Control, 0.25, 0.35 and 0.45g/kg of butyric acid) under completely randomized design. Each treatment was replicated 3 times with 28 birds in each replicate. Feed intake, body weight gain, feed conversion ratio, intestinal morphology, apparent ileal digestibility of protein and immunity parameters were evaluated. At the end of the experiment (35-d) 3 birds/replicate in each group were randomly selected and slaughtered to collect blood, duodenal samples and ileal digesta. The data were analyzed by using ANOVA technique. The results indicated improved body weight gain (P = 0.0222), feed conversion ratio (P = 0.0056), duodenal villus height (P = 0.0512), AID (P = 0.0098) antibody titer against Newcastle disease improved (P = 0.0326). Treatments remained unresponsive with respect to feed intake (P = 0.9685).

Keywords: butyric acid, broilers, gut health, ileal digestibility

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Authors: Madhuresh Dwivedi, Navneet Singh Deora, H. N. Mishra

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In the context of bakery products, the gluten component of wheat has a crucial role in stabilizing the gas-cell and crumb structures, appearance, mouth feel and maintaining the rheological properties, thus the acceptability of these products. However, because of coeliac disease, some individuals cannot tolerate the protein gliadin present in the gluten fraction of wheat flour. Also termed as gluten-sensitive enteropathy, it is a common chronicle disorder in populations throughout the world with average prevalence of 0.37%. The safest way for celiac sufferers is to stay away from gluten-containing foods such as wheat, rye, barley as well as durum wheat, spelt wheat, and triticale. Thus, in view of the current increasing incidence of gluten intolerant sufferers (due to improved diagnostic procedures), the development of gluten-free cereal-based bakery products suitable for celiac patients represents a challenging and serious task, but also very demanding call for food technologists as well as for the bakers. The use of alternative cereal starches (like rice, soy, maize, potato and so on), gums, hydrocolloids, dietary fibres, alternative protein sources, prebiotics and combinations of them represent the most widespread approach used as replacement to mimic gluten in the manufacture of industrial processable gluten-free bakery products due to their structure-building and water binding properties.

Keywords: gluten-free, coeliac disease, alternative flour, hydrocolloid, crumb structure

Procedia PDF Downloads 275