Search results for: milk protein concentrate

Authors: Harish Kumar Sharma

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The influence of different levels of inulin as a fat replacer on the quality of ice cream was investigated. The physicochemical, rheological and textural properties of control ice cream and ice cream prepared with inulin in different proportions were determined and correlated to the different parameters using Pearson correlation and Principle Component Analysis (PCA). Based on the overall acepectability, ice cream with 4% inulin was found best and was selected for preparation of ice cream with inulin:SPI in different proportions. Compared with control ice cream, Inulin:SPI showed different rheological properties, resulting in significantly higher apparent viscosities, consistency coefficient and greater deviations from Newtonian flow. In addition, both hardness and melting resistance significantly increased with increase in the SPI content in ice cream prepared with inulin: SPI. Also hardness value increased for inulin based ice cream compared to control ice cream but it melted significantly faster than the latter. Colour value significantly decreased in both the cases compared to the control sample. The deliberation shall focus on the effect of incorporation of inulin on the quality of ice-cream.

Keywords: fat replacer, inulin, ice cream, viscosity, principal component analysis

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Authors: Ruwani N. Nugara, Masashi Inafuku, Kensaku Takara, Hironori Iwasaki, Hirosuke Oku

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Pteryxin from the partially purified hexane phase (HP) of Peucedanum japonicum Thunb (PJT) was identified as the active compound related to anti-obesity. Thus, in this study we investigated the mechanisms related to anti-obesity activity in-vitro. The HP was fractionated, and effect on the triglyceride (TG) content was evaluated in 3T3-L1 and HepG2 cells. Comprehensive spectroscopic analyses were used to identify the structure of the active compound. The dose dependent effect of active constituent on the TG content, and the gene expressions related to adipogenesis, fatty acid catabolism, energy expenditure, lipolysis and lipogenesis (20 μg/mL) were examined in-vitro. Furthermore, higher dosage of pteryxin (50μg/mL) was tested against 20μg/mL in 3T3-L1 adipocytes. The mRNA were subjected to SOLiD next generation sequencer and the obtained data were analyzed by Ingenuity Pathway Analysis (IPA). The active constituent was identified as pteryxin, a known compound in PJT. However, its biological activities against obesity have not been reported previously. Pteryxin dose dependently suppressed TG content in both 3T3-L1 adipocytes and HepG2 hepatocytes (P < 0.05). Sterol regulatory element-binding protein-1 (SREBP1 c), Fatty acid synthase (FASN), and acetyl-CoA carboxylase-1 (ACC1) were downregulated in pteryxin-treated adipocytes (by 18.0, 36.1 and 38.2%; P < 0.05, respectively) and hepatocytes (by 72.3, 62.9 and 38.8%, respectively; P < 0.05) indicating its suppressive effects on fatty acid synthesis. The hormone-sensitive lipase (HSL), a lipid catabolising gene was upregulated (by 15.1%; P < 0.05) in pteryxin-treated adipocytes suggesting improved lipolysis. Concordantly, the adipocyte size marker gene, paternally expressed gene1/mesoderm specific transcript (MEST) was downregulated (by 42.8%; P < 0.05), further accelerating the lipolytic activity. The upregulated trend of uncoupling protein 2 (UCP2; by 77.5%; P < 0.05) reflected the improved energy expenditure due to pteryxin. The 50μg/mL dosage of pteryxin completely suppressed PPARγ, MEST, SREBP 1C, HSL, Adiponectin, Fatty Acid Binding Protein (FABP) 4, and UCP’s in 3T3-L1 adipocytes. The IPA suggested that pteryxin at 20μg/mL and 50μg/mL suppress obesity in two different pathways, whereas the WNT signaling pathway play a key role in the higher dose of pteryxin in preadipocyte stage. Pteryxin in PJT play the key role in regulating lipid metabolism related gene network and improving energy production in vitro. Thus, the results suggests pteryxin as a new natural compound to be used as an anti-obesity drug in pharmaceutical industry.

Keywords: obesity, peucedanum japonicum thunb, pteryxin, food science

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Authors: Jinyoung Park, Eun Joo Song

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Introduction: Deubiquitinating enzymes (DUBs) are proteases that cleave ubiquitin or ubiquitin-like modifications on substrates. Deubiquitination could regulate cellular physiology, such as signal transduction, DNA damage and repair, and cell cycle progression. Although more than 100 DUBs are encoded in the human and the importance of DUBs has been realized, the functions of most DUBs are unknown. This study aims to identify the molecular mechanism by which deubiquitinating enzyme USP35 regulates cell cycle progression for the first time. Methods: USP35 RNAi was mainly used to identify the function of USP35 in cell cycle progression. To find substrates of USP35, we analyzed protein-protein interaction using LC-MS. Several biological methods, such as ubiquitination assay, cell synchronization, immunofluorescence, and immunoprecipitation assay were used to investigate the exact mechanism by which USP35 affects successful completion of mitosis. Results: USP35 knockdown caused not only reduction of mitotic cell number but also induction of mitotic cells with abnormal spindle formation. Actually, cell proliferation was decreased by USP35 knockdown. Interestingly, we found that loss of USP35 decreased the stability and expression of Aurora B, a member of chromosomal passenger complex (CPC), and the phosphorylation of its substrate. Indeed, USP35 interacted with Aurora B and deubiquitinated it. In addition, USP35 knockdown induced abnormal localization of Aurora B in mitotic cells. Finally, CDH1-mediated ubiquitination of Aurora B level was rescued by USP35 overexpression, but not inactive form of USP35, USP35 C450A. Discussion: Our findings suggest that USP35 regulates Aurora B-mediated mitotic spindle assembly and G2-M transition by blocking CDH1-induced degradation of Aurora B.

Keywords: USP35, HSP90, Aurora B, cell cycle progression

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Authors: S. Hussain, N. Ahmad, S. Alam, M. Bezabhi, W. H. Hendriks, P. Yu, J. W. Cone

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The high content of lignin in cell walls is the major limiting factor in the digestion and utilisation of cereal crop residues by ruminants. The aim of this study was to evaluate the effectiveness of the white rot fungus, Pleurotus ostreatus (P. ostreatus), to degrade lignin and to enhance the rumen degradability of maize stover, rice straw, wheat straw and their mixture in equal proportion on a dry-matter (DM) basis. Four samples of each substrate were incubated aerobically in triplicate with P. ostreatus for 0 (Control), 21, 28 and 35 days under solid-state conditions (temperature, 24 ͦ C; humidity, 70± 5%). The changes in chemical composition, DM and nutrient losses, and rumen fermentation characteristics using in vitro DM digestibility (DMD) and the in vitro gas production (GP) technique were measured. The results showed that incubation with P. ostreatus decreased (P < 0.001) the contents of neutral detergent fibre and lignin with a concomitant increase (P < 0.001) in the contents of ash and crude protein. The losses of nutrients differed (P < 0.001) among the straw types, with rice straw and maize stover showing the largest (P < 0.05) lignin degradation compared to wheat and mixed straws. The DMD and 72-h cumulative GP increased (P < 0.001) consistently with increasing fungal incubation period and for all substrates the highest values of DMD and GP were measured after 35 days of incubation with P. ostreatus. The lignin degradation was strongly associated with hemicellulose degradation (r = 0.71) across the various straws. Results of the present study demonstrated that incubation of low-quality crop residues with P. ostreatus under solid-state conditions upgrades their feeding value by reducing the content of lignin and increasing the content of crude protein and ruminal degradation.

Keywords: crop residues, lignin degradation, maize stovers, wheat straws, white rot fungi

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Authors: Monika Kallubai, Shreya Dubey, Rajagopal Subramanyam

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Our study is all about the biological interactions of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules with carrier protein Human Serum Albumin (HSA). The cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 µM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. The further experiment proved that Dhc and DhcA induced 35.6% and 37.7% early apoptotic cells and 2.5%, 2.9% late apoptotic cells respectively. Morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7±0.03×104 M-1 and 3.9±0.05×104 M-1, and their binding free energies were found to be -6.4 and -6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4±0.05×104 M-1 replaced Dhc, and phenylbutazone 1.5±0.05×104 M-1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular sizes of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported the greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for the further development of steroid derivatives with improved pharmacological significance as novel anti-cancer drugs.

Keywords: apoptosis, dihydrocortisol, fluorescence quenching, protein conformations

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Authors: Shigdaf Mekuriaw, Firew Tegegne, A. Tsunekawa, Dereje Tewabe

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The purpose of this paper is to make a comprehensive review on the use of water hyacinth (Eichhornia crassipes) as a potential livestock feed and argue its utilization as complementary strategy to other control methods. Water Hyacinth is one of the most noxious plant invaders of rivers and lakes. Such weeds cause environmental disaster and interfere with economic and recreational activities such as water transportation and fishing. Economic impacts of the weed in seven African countries have been estimated at between 20-50 million US$ every year. It would, therefore, be prudent to suggest utilization as a complementary control method. The majority of people in developing countries are dependent on traditional and inefficient crop-livestock production system that constrains their ability to enhance economic productivity and quality of life. Livestock in developing countries faces shortage of feed, especially during the long dry seasons. Existing literature shows the use of water hyacinth as livestock and fish feed. The chemical composition of water hyacinth varies considerably. Due to its relatively high crude protein (CP) content (5.8-20.0%), water hyacinth can be considered as a potential protein supplement for livestock which commonly feed cereal crop residues whose contribution as source of feed is increasing in Africa. Though the effects of anti-nutritional factors (ANFs) present in water hyacinth is not investigated, their concentrations are not above threshold hinder its utilization as livestock feed. In conclusion, water hyacinth could provide large quantities of nutritious feed for animals. Like other feeds, water hyacinth may not be offered as a sole feed and based on existing literature its optimum inclusion level reaches 50%.

Keywords: Africa, livestock feed, water bodies, water hyacinth and weed control method

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Authors: Nicole C. Valdez, Vincent L. Borromeo, Conrad C. Chong, Ahmad F. Mazahery

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Breast cancer is the most prevalent cancer worldwide, where the majority of cases are estrogen-receptor positive and involve 2 receptor proteins. The binding of estrogen to estrogen receptor alpha (ERα) promotes breast cancer growth, while it's binding to estrogen-receptor beta (ERβ) inhibits tumor growth. While natural products have been a promising source of chemotherapeutic agents, the challenge remains in finding a bioactive compound that specifically targets cancer cells, minimizing side effects on normal cells. Flavonoids are natural products that act as phytoestrogens and induce the same response as estrogen. They are able to compete with estrogen for binding to ERα; however, it has a higher binding affinity for ERβ. Their abundance in nature and low toxicity make them a potential candidate for breast cancer treatment. This study aimed to determine which particular flavonoids can specifically recognize ERβ and potentially be used for breast cancer treatment through molecular docking. A total of 206 flavonoids comprised of 97 isoflavones and 109 flavanones were collected from ZINC15, while the 3D structures of ERβ and ERα were obtained from Protein Data Bank. These flavonoid subclasses were chosen as they bind more strongly to ERs due to their chemical structure. The structures of the flavonoid ligands were converted using Open Babel, while the estrogen receptor protein structures were prepared using Autodock MGL Tools. The optimal binding site was found using BIOVIA Discovery Studio Visualizer before docking all flavonoids on both ERβ and ERα through Autodock Vina. Genistein is a flavonoid that exhibits anticancer effects by binding to ERβ, so its binding affinity was used as a baseline. Eriodictyol and 4”,6”-Di-O-Galloylprunin both exceeded genistein’s binding affinity for ERβ and was lower than its binding affinity for ERα. Of the two, eriodictyol was pursued due to its antitumor properties on a lung cancer cell line and on glioma cells. It is able to arrest the cell cycle at the G2/M phase by inhibiting the mTOR/PI3k/Akt cascade and is able to induce apoptosis via the PI3K/Akt/NF-kB pathway. Protein pathway and gene analysis were also conducted using ChEMBL and PANTHER and it was shown that eriodictyol might induce anticancer effects through the ROS1, CA7, KMO, and KDM1A genes which are involved in cell proliferation in breast cancer, non-small cell lung cancer, and other diseases. The high binding affinity of eriodictyol to ERβ, as well as its potential affected genes and antitumor effects, therefore, make it a candidate for the development of new breast cancer treatment. Verification through in vitro experiments such as checking the upregulation and downregulation of genes through qPCR and checking cell cycle arrest using a flow cytometry assay is recommended.

Keywords: breast cancer, estrogen receptor, flavonoid, molecular docking

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Authors: Mohamed A. Morsy, Azza A. El-Sheikh, Abdulla Y. Al-Taher

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Resveratrol (RES) is a well-known polyphenol antioxidant. We have previously shown that testicular protective effect of RES against the anticancer drug methotrexate (MTX)-induced toxicity involves transporter-mediated mechanisms. Here, we investigated the effect of RES on MTX-induced nephrotoxicity. Rats were administered RES (10 mg/kg/day) for 8 days, with or without a single MTX dose (20 mg/kg i.p.) at day 4 of the experiment. MTX induced nephrotoxicity evident by significantly increase in serum blood urea nitrogen and creatinine compared to control, as well as distortion of kidney microscopic structure. MTX also significantly increased renal nitric oxide level, with induction of inducible nitric oxide synthase expression. MTX also significantly up-regulated fas ligand and caspase 3. Administering RES prior to MTX significantly improved kidney function and microscopic picture, as well as significantly decreased nitrosative and apoptotic markers compared to MTX alone. RES, but not MTX, caused significant increase in expression of breast cancer resistance protein (BCRP), an apical efflux renal transporter that participates in urinary elimination of both MTX and RES. Interestingly, concomitant MTX and RES caused further up-regulation of renal Bcrp compared to RES alone. Using Human BCRP ATPase assay, both RES and MTX exhibited dose-dependent increase in ATPase activity, with Km values of 0.52 ± 0.03 and 30.9 ± 4.2 µM, respectively. Furthermore, combined RES and MTX caused ATPase activity which was significantly less than maximum ATPase activity attained by the positive control; sulfasalazine (12.5 µM). In conclusion, RES exerted nephro-protection against MTX-induced toxicity through anti-nitrosative and anti-apoptotic effects, as well as via up-regulation of renal Bcrp.

Keywords: methotrexate, resveratrol, nephrotoxicity, breast cancer resistance protein

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Authors: Moheb R. Girgis, Tarek M. Mahmoud, Bahgat A. Abdullatif, Ahmed M. Rabie

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Mesh clients, mesh routers and gateways are components of Wireless Mesh Network (WMN). In WMN, gateways connect to Internet using wireline links and supply Internet access services for users. We usually need multiple gateways, which takes time and costs a lot of money set up, due to the limited wireless channel bit rate. WMN is a highly developed technology that offers to end users a wireless broadband access. It offers a high degree of flexibility contrasted to conventional networks; however, this attribute comes at the expense of a more complex construction. Therefore, a challenge is the planning and optimization of WMNs. In this paper, we concentrate on this challenge using a genetic algorithm and simulated annealing. The genetic algorithm and simulated annealing enable searching for a low-cost WMN configuration with constraints and determine the number of used gateways. Experimental results proved that the performance of the genetic algorithm and simulated annealing in minimizing WMN network costs while satisfying quality of service. The proposed models are presented to significantly outperform the existing solutions.

Keywords: wireless mesh networks, genetic algorithms, simulated annealing, topology design

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Authors: Austin Jiang, Richard M. Salmon, Nicholas W. Morrell, Wei Li

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BMP10 is highly expressed in the developing heart and plays essential roles in cardiogenesis. BMP10 deletion in mice results in embryonic lethality due to impaired cardiac development. In adults, BMP10 expression is restricted to the right atrium, though ventricular hypertrophy is accompanied by increased BMP10 expression in a rat hypertension model. However, reports of BMP10 activity in the circulation are inconclusive. In particular it is not known whether in vivo secreted BMP10 is active or whether additional factors are required to achieve its bioactivity. It has been shown that high-affinity binding of the BMP10 prodomain to the mature ligand inhibits BMP10 signaling activity in C2C12 cells, and it was proposed that prodomain-bound BMP10 (pBMP10) complex is latent. In this study, we demonstrated that the BMP10 prodomain did not inhibit BMP10 signaling activity in multiple endothelial cells, and that recombinant human pBMP10 complex, expressed in mammalian cells and purified under native conditions, was fully active. In addition, both BMP10 in human plasma and BMP10 secreted from the mouse right atrium were fully active. Finally, we confirmed that active BMP10 secreted from mouse right atrium was in the prodomain-bound form. Our data suggest that circulating BMP10 in adults is fully active and that the reported vascular quiescence function of BMP10 in vivo is due to the direct activity of pBMP10 and does not require an additional activation step. Moreover, being an active ligand, recombinant pBMP10 may have therapeutic potential as an endothelial-selective BMP ligand, in conditions characterized by loss of BMP9/10 signaling.

Keywords: bone morphogenetic protein 10 (BMP10), endothelial cell, signal transduction, transforming growth factor beta (TGF-B)

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Authors: Pravinchandra Patel

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Sustainable agriculture in sandy loam soil generally faces large constraints due to low water holding and nutrient retention capacity, and accelerated mineralization of soil organic matter. There is need to increase soil organic carbon in the soil for higher crop productivity and soil sustainability. Recently biochar is considered as sixth element and work as a catalyst for increasing crop yield, soil fertility, soil sustainability and mitigation of climate change. Biochar was generated at the Sansoli Farm of Anand Agricultural University, Gujarat, India by pyrolysis at temperatures (250-400°C) in absence of oxygen using slow chemical process (using two kilns) from corn stover (Zea mays, L), cluster bean stover (Cyamopsis tetragonoloba) and Prosopis julifera wood. There were 16 treatments; 4 organic sources (3 biochar; corn stover biochar (MS), cluster bean stover (CB) & Prosopis julifera wood (PJ) and one farmyard manure-FYM) with two rate of application (5 & 10 metric tons/ha), so there were eight treatments of organic sources. Eight organic sources was applied with the recommended dose of fertilizers (RDF) (80-40-0 kg/ha N-P-K) while remaining eight organic sources were kept without RDF. Application of corn stover biochar @ 10 metric tons/ha along with RDF (RDF+MS) increased dry matter (DM) yield, crude protein (CP) yield, chlorophyll content and plant height (at 30 and 60 days after sowing) than CB and PJ biochar and FYM. Nutrient uptake of P, K, Ca, Mg, S and Cu were significantly increased with the application of RDF + corn stover @ 10 metric tons/ha while uptake of N and Mn were significantly increased in RDF + corn stover @ 5 metric tons/ha. It was found that soil application of corn stover biochar @ 10 metric tons/ha along with the recommended dose of chemical fertilizers (RDF+MS ) exhibited the highest impact in obtaining significantly higher dry matter and crude protein yields and larger removal of nutrients from the soil and it also beneficial for built up nutrients in soil. It also showed significantly higher organic carbon content and cation exchange capacity in sandy loam soil. The lower dose of corn stover biochar @ 5 metric tons/ha (RDF+ MS) was also remained the second highest for increasing dry matter and crude protein yields of forage corn crop which ultimately resulted in larger removals of nutrients from the soil. This study highlights the importance of mixing of biochar along with recommended dose of fertilizers on its synergistic effect on sandy loam soil nutrient retention, organic carbon content and water holding capacity hence, the amendment value of biochar in sandy loam soil.

Keywords: biochar, corn yield, plant nutrient, fertility status

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Authors: Majede Modheji, Hamid Emadi, Hossein Vojoudi

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An efficient magnetic mesoporous structure was designed and prepared for the facile pre-concentration of As(III) ions. To prepare the sorbent, a core-shell magnetic silica nanoparticle was covered by MCM-41 like structure, and then the surface was modified by guanidine via an amine linker. The prepared adsorbent was investigated as an effective and sensitive material for the adsorption of arsenic ions from the aqueous solution applying a normal batch method. The imperative variables of the adsorption were studied to increase efficiency. The dynamic and static processes were tested that matched a pseudo-second order of kinetic model and the Langmuir isotherm model, respectively. The sorbent reusability was investigated, and it was confirmed that the designed product could be applied at best for six cycles successively without any significant efficiency loss. The synthesized product was tested to determine and pre-concentrate trace amounts of arsenic ions in rice and natural waters as a real sample. A desorption process applying 5 mL of hydrochloric acid (0.5 mol L⁻¹) as an eluent exhibited about 98% recovery of the As(III) ions adsorbed on the GA-MSMP sorbent.

Keywords: arsenic, adsorption, mesoporous, surface modification, MCM-41

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Authors: Juliana Lukša, Bazilė Ravoitytė, Elena Servienė, Saulius Serva

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Totiviridae L-A virus is a persistent Saccharomyces cerevisiae dsRNA virus. It encodes the major structural capsid protein Gag and Gag-Pol fusion protein, responsible for virus replication and encapsulation. These features also enable the copying of satellite dsRNAs (called M dsRNAs) encoding a secreted toxin and immunity to it (known as killer toxin). Viral capsid pore presumably functions in nucleotide uptake and viral mRNA release. During cell division, sporogenesis, and cell fusion, the virions remain intracellular and are transferred to daughter cells. By employing high throughput RNA sequencing data analysis, we describe the influence of solely L-A virus on the expression of genes in three different S. cerevisiae hosts. We provide a new perception into Totiviridae L-A virus-related transcriptional regulation, encompassing multiple bioinformatics analyses. Transcriptional responses to L-A infection were similar to those induced upon stress or availability of nutrients. It also delves into the connection between the cell metabolism and L-A virus-conferred demands to the host transcriptome by uncovering host proteins that may be associated with intact virions. To better understand the virus-host interaction, we applied differential proteomic analysis of virus particle-enriched fractions of yeast strains that harboreither complete killer system (L-A-lus and M-2 virus), M-2 depleted orvirus-free. Our analysis resulted in the identification of host proteins, associated with structural proteins of the virus (Gag and Gag-Pol). This research was funded by the European Social Fund under the No.09.3.3-LMT-K-712-19-0157“Development of Competences of Scientists, other Researchers, and Students through Practical Research Activities” measure.

Keywords: totiviridae, killer virus, proteomics, transcriptomics

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Authors: Miguel Pereira, Lígia Pimentel, Manuela Pintado

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Animal blood is a major by-product of slaughterhouses and still represents a cost and environmental problem in some countries. To be eliminated, blood should be stabilised by cooking and afterwards the slaughterhouses must have to pay for its incineration. In order to reduce the elimination costs and valorise the high protein content the aim of this study was the optimization of hydrolysis conditions, in terms of enzyme ratio and time, in order to obtain hydrolysates with biological activity. Two enzymes were tested in this assay: pepsin and proteases from Cynara cardunculus (cardosins). The latter has the advantage to be largely used in the Portuguese Dairy Industry and has a low price. The screening assays were carried out in a range of time between 0 and 10 h and using a ratio of enzyme/reaction volume between 0 and 5%. The assays were performed at the optimal conditions of pH and temperature for each enzyme: 55 °C at pH 5.2 for cardosins and 37 °C at pH 2.0 for pepsin. After reaction, the hydrolysates were evaluated by FPLC (Fast Protein Liquid Chromatography) and tested for their antioxidant activity by ABTS method. FPLC chromatograms showed different profiles when comparing the enzymatic reactions with the control (no enzyme added). The chromatogram exhibited new peaks with lower MW that were not present in control samples, demonstrating the hydrolysis by both enzymes. Regarding to the antioxidant activity, the best results for both enzymes were obtained using a ratio enzyme/reactional volume of 5% during 5 h of hydrolysis. However, the extension of reaction did not affect significantly the antioxidant activity. This has an industrial relevant aspect in what concerns to the process cost. In conclusion, the enzymatic blood hydrolysis can be a better alternative to the current elimination process allowing to the industry the reuse of an ingredient with biological properties and economic value.

Keywords: antioxidant activity, blood, by-products, enzymatic hydrolysis

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Authors: Seyed-Ahmad Shahidi, Salemeh Kazemzadeh, Mehdi Sharifi Soltani, Azade Ghorbani-HasanSaraei

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The kinetics of colour changes of American Pokeweed extract, due to concentration by various heating methods was studied. Three different heating/evaporation processes were employed for production of American Pokeweed extract concentrate. The American Pokeweed extract was concentrated to a final 40 °Brix from an initial °Brix of 4 by microwave heating, rotary vacuum evaporator and evaporating at atmospheric pressure. The final American Pokeweed extract concentration of 40 °Brix was achieved in 188, 216 and 320 min by using microwave, rotary vacuum and atmospheric heating processes, respectively. The colour change during concentration processes was investigated. Total colour differences, Hunter L, a and b parameters were used to estimate the extent of colour loss. All Hunter colour parameters decreased with time. The zero-order, first-order and a combined kinetics model were applied to the changes in colour parameters. All models were found to describe the L, a and b-data adequately. Results indicated that variation in TCD followed both first-order and combined kinetics models. This model implied that the colour formation and pigment destruction occurred during concentration processes of American Pokeweed extract.

Keywords: American pokeweed, colour, concentration, kinetics

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Authors: Kanchana Sitlaothaworn

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Yogurt capsule was made by mixing 14% w/v of reconstitution of skim milk with 2% FOS. The mixture was fermented by commercial yogurt starter comprising Lactobacillus bulgaricus and Streptococcus thermophilus. These yogurts were made as yogurt powder by freeze-dried. Yogurt powder was put into capsule then stored for 28 days at 4oc. 8ml of commercial yogurt was found to be the most suitable inoculum size in yogurt production. After freeze-dried, the viability of L. bulgaricus and S. thermophilus reduced from 109 to 107 cfu/g. The precence of sucrose cannot help to protect cell from ice crystal formation in freeze-dried process, high (20%) sucrose reduced L. bulgaricus and S. thermophilus growth during fermentation of yogurt. The addition of FOS had reduced slowly the viability of both L. bulgaricus and S. thermophilus similar to control (without FOS) during 28 days of capsule storage. The viable cell exhibited satisfactory viability level in capsule storage (6.7x106cfu/g) during 21 days at 4oC.

Keywords: yogurt capsule, Lactobacillus bulgaricus, Streptococcus thermophilus, freeze-drying, sucrose

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Authors: Y. Boulaksil, J. C. Fransoo, E.E. Blanco, S. Koubida

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In this paper, we study the small traditional retailers that are located in the neighborhoods of big cities in emerging markets. Although modern retailing has grown in the last two decades in these markets, the number of small retailers is still increasing and serving a substantial part of the daily demand for many basic products, such as bread, milk, and cooking oil. We conduct an empirical study to understand the business environment of these small traditional retailers in emerging markets by collecting data from 333 small retailers, spread over 8 large cities in Morocco. We analyze the data and describe their business environment with a focus on the informal credits they offer to their customers. We find that smaller small retailers that are funded from personal savings and managed by the owner himself offer relatively the most credits. Our study also provides interesting insights about these small retailers that will help FMCG manufacturers that are (planning to be) active in Morocco and other emerging markets. We also discuss a number opportunities to improve the efficiency of the supply chains that serve them.

Keywords: small retailers, big cities, emerging markets, empirical study, supply chain management, Morocco

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Authors: B. T. Naveen Kumar, Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, A. H. Shanthanagouda, Orawan Boodde, K. M. Shankar, Prakash Patil, Shubhkaramjeet Kaur

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Early mortality syndrome (EMS)/Acute Hepatopancreatic Necrosis Disease (AHPND) has emerged as a major obstacle for the shrimp farming around the world. It is caused by a strain of Vibrio parahaemolyticus. The possible preventive and control measure is, early and rapid detection of the pathogen in the broodstock, post-larvae and monitoring the shrimp during the culture period. Polymerase chain reaction (PCR) based early detection methods are good, but they are costly, time taking and requires a sophisticated laboratory. The present study was conducted to develop a simple, sensitive and rapid diagnostic farmer level kit for the reliable detection of AHPND in shrimp. A panel of monoclonal antibodies (MAbs) were raised against the recombinant Pir B protein (rPirB). First, an immunodot was developed by using MAbs G3B8 and Mab G3H2 which showed specific reactivity to purified r-PirB protein with no cross-reactivity to other shrimp bacterial pathogens (AHPND free Vibrio parahaemolyticus (Indian strains), V. anguillarum, WSSV, Aeromonas hydrophila, and Aphanomyces invadans). Immunodot developed using Mab G3B8 is more sensitive than that with the Mab G3H2. However, immunodot takes almost 2.5 hours to complete with several hands-on steps. Therefore, the flow-through assay (FTA) was developed by using a plastic cassette containing the nitrocellulose membrane with absorbing pads below. The sample was dotted in the test zone on the nitrocellulose membrane followed by continuos addition of five solutions in the order of i) blocking buffer (BSA) ii) primary antibody (MAb) iii) washing Solution iv) secondary antibody and v) chromogen substrate (TMB) clear purple dots against a white background were considered as positive reactions. The FTA developed using MAbG3B8 is more sensitive than that with MAb G3H2. In FTA the two MAbs showed specific reactivity to purified r-PirB protein and not to other shrimp bacterial pathogens. The FTA is simple to farmer/field level, sensitive and rapid requiring only 8-10 min for completion. Tests can be developed to kits, which will be ideal for use in biosecurity, for the first line of screening (at the port or pond site) and during monitoring and surveillance programmes overall for the good management practices to reduce the risk of the disease.

Keywords: acute hepatopancreatic necrosis disease, AHPND, flow-through assay, FTA, farmer level, immunodot, pond site, shrimp

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Authors: Jamaa Salih, Francis Edum-Fotwe, Andrew Price

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One of the most critical aspects of the management of road infrastructure is the type and scale of maintenance systems adopted and the consequences of their inadequacy. The performance of road maintenance systems can be assessed by a number of important indicators such as: cost, safety, environmental impact, and level of complaints by users. A review of practice reveals that insufficient level of expenditure or poor management of the road network often has serious consequences for the economic and social life of a country in terms of vehicle operating costs (VOC), travel time costs, accident costs and environmental impact. Despite an increase in the attention paid by global road agencies to the environmental and the road users’ satisfaction, the overwhelming evidence from the available literature agree on the lack of similar levels of attention for the two factors in many developing countries. While many sources agree that the road maintenance backlog is caused by either the shortage of expenditures or lack of proper management or both, it appears that managing the available assets particularly in the developing countries is the main issue. To address this subject, this paper will concentrate on exposing the various issues related to this field.

Keywords: environmental impact, performance indicators, road maintenance, users’ satisfaction

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Authors: Hyun Lim, Dong Sook Min, Han Eul Yun, Kil Tae Kim, Ya Nan Sun, Young Ho Kim, Hyun Pyo Kim

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Matrix metalloproteinase (MMP)-13 has an important role for degrading cartilage materials under inflammatory conditions such as arthritis. Since the 70% ethanol extract of Acanthopanax koreanum inhibited MMP-13 expression in IL-1β-treated human chondrocyte cell line, SW1353, two major constituents including acanthoic acid and impressic acid were initially isolated from the same plant materials and their MMP-13 down-regulating capacity was examined. In IL-1β-treated SW1353 cells, acanthoic acid and impressic acid significantly and concentration-dependently inhibited MMP-13 expression at 10 – 100 μM and 0.5 – 10 μM, respectively. The potent one, impressic acid, was found to inhibit MMP-13 expression by blocking the phosphorylation of signal transducer and activator of transcription-1/-2 (STAT-1/-2) and activation of c-Jun and c-Fos among cellular signaling pathway involved, but did not affect the activation of mitogen-activated protein kinases (MAPKs) and nuclear transcription factor-κB (NF-κB). Further, impressic acid was also found to inhibit the expression of MMP-13 mRNA (47.7% inhibition at 10 μM), the glycosaminoglycan release (42.2% reduction at 10 μM) and proteoglycan loss in IL-1-treated rabbit cartilage explants culture. For a further study, 21 impressic acid derivatives were isolated from the same plant materials and their suppressive activities against MMP-13 expression were examined. Among the derivatives, 3α-hydroxy-lup-20(29)-en-23-oxo,28-oic acid, (20R)-3α-hydroxy-29-dimethoxylupan-23,28-dioic acid, acankoreoside F and acantrifoside A clearly down-regulated MMP-13 expression, but impressic acid being most potent. All these results suggest that impressic acid, 3α-hydroxy-lup-20(29)-en-23-oxo,28-oic acid, (20R)-3α-hydroxy-29-dimethoxylupan-23,28-dioic acid, acankoreoside F, acantrifoside A and A. koreanum may have a potential for therapeutic agents to prevent cartilage degradation possibly by inhibiting matrix protein degradation.

Keywords: acanthoic acid, Acanthopanax koreanum, cartilage, impressic acid, matrix metalloproteinase

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Authors: R. Razavizadeh, F. Adabavazeh, M. Rezaee Chermahini

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Salinity is one of the most widespread agricultural problems in arid and semi-arid areas that limits the plant growth and crop productivity. In this study, the salt stress effects on protein, reducing sugar, proline contents and antioxidant enzymes activities of Carum copticum L. under in vitro conditions were studied. Seeds of C. copticum were cultured in Murashige and Skoog (MS) medium containing 0, 25, 50, 100 and 150 mM NaCl and calli were cultured in MS medium containing 1 μM 2, 4-dichlorophenoxyacetic acid, 4 μM benzyl amino purine and different levels of NaCl (0, 25, 50, 100 and 150 mM). After NaCl treatment for 28 days, the proline and reducing sugar contents of shoots, roots and calli increased significantly in relation to the severity of the salt stress. The highest amount of proline and carbohydrate were observed at 150 and 100 mM NaCl, respectively. The reducing sugar accumulation in shoots was the highest as compared to roots, whereas, proline contents did not show any significant difference in roots and shoots under salt stress. The results showed significant reduction of protein contents in seedlings and calli. Based on these results, proteins extracted from the shoots, roots and calli of C. copticum treated with 150 mM NaCl showed the lowest contents. The positive relationships were observed between activity of antioxidant enzymes and the increase in stress levels. Catalase, ascorbate peroxidase and superoxide dismutase activity increased significantly under salt concentrations in comparison to the control. These results suggest that the accumulation of proline and sugars, and activation of antioxidant enzymes play adaptive roles in the adaptation of seedlings and callus of C. copticum to saline conditions.

Keywords: antioxidant enzymes, Carum copticum, organic solutes, salt stress

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Authors: Dong Tran, Thanh Dac Van, Ly Le

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Hemagglutinin (HA) and Neuraminidase (NA) play an important role in host immune evasion across influenza virus evolution process. The correlation between HA and NA evolution in respect to epitopic evolution and drug interaction has yet to be investigated. In this study, combining of sequence to structure evolution and statistical analysis on epitopic/binding site specificity, we identified potential therapeutic features of HA and NA that show specific antibody binding site of HA and specific binding distribution within NA active site of current inhibitors. Our approach introduces the use of sequence variation and molecular interaction to provide an effective strategy in establishing experimental based distributed representations of protein-protein/ligand complexes. The most important advantage of our method is that it does not require complete dataset of complexes but rather directly inferring feature interaction from sequence variation and molecular interaction. Using correlated sequence analysis, we additionally identified co-evolved mutations associated with maintaining HA/NA structural and functional variability toward immunity and therapeutic treatment. Our investigation on the HA binding specificity revealed unique conserved stalk domain interacts with unique loop domain of universal antibodies (CR9114, CT149, CR8043, CR8020, F16v3, CR6261, F10). On the other hand, NA inhibitors (Oseltamivir, Zaninamivir, Laninamivir) showed specific conserved residue contribution and similar to that of NA substrate (sialic acid) which can be exploited for drug design. Our study provides an important insight into rational design and identification of novel therapeutics targeting universally recognized feature of influenza HA/NA.

Keywords: influenza virus, hemagglutinin (HA), neuraminidase (NA), sequence evolution

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Authors: Igor Sukhotnik

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Objectives: Notch signaling is thought to act to drive cell versification in the lining of the small intestine. When Notch signaling is blocked, proliferation ceases, and epithelial cells become secretory. The purpose of the present study was to evaluate the role of Notch signaling pathway in stem cell differentiation in a rat model of intestinal ischemia-reperfusion (IR). Methods: Male Sprague-Dawley rats were randomly divided into four experimental groups: Sham-24 and Sham-48 rats underwent laparotomy and were killed 24 or 48 h later, respectively; IR-24 and IR-48 rats underwent occlusion of SMA and portal vein for 30 min followed by 24 or 48 h of reperfusion, respectively. Notch-related gene and protein expression were determined using Real Time PCR, Western blotting and immunohistochemistry. Wax histology and immunohistochemistry was used to determine cell differentiation toward absorptive (enterocytes) or secretory progenitors (goblet cells, enteroendocrine cells or Paneth cells). Results: IR-48 rats exhibited a significant decrease in Notch-1 protein expression (Western blot) that was coincided with a significant decrease in the number of Notch-1 positive cells (immunohistochemistry) in jejunum and ileum as well as Hes-1 positive cells in jejunum and ileum compared to Sham-48 rats. A significant down-regulation of Notch signaling related genes and proteins in IR animals was accompanied by a significant increase in the number of goblet and Paneth cells and decreased number of absorptive cells compared to control rats. Conclusions: Forty-eight hours following intestinal IR in rats, inhibited Notch signaling pathway was accompanied by intestinal stem cells differentiation toward secretory progenitors.

Keywords: Intestine, notch, ischemia-reperfusion, cell differentiation, secretory

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Authors: Yu-Ting Chen, Shiao-Shing Chen, Hung-Te Hsu, Saikat Sinha Ray

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Forward osmosis (FO) is a crucial technology with low operating pressure and cost for water reuse and reclamation. In Taiwan, with the advance of science and technology, thin film transistor liquid crystal displays (TFT-LCD) based industries are growing exponentially. In the optoelectronic industry wastewater, the iodide is one of the valuable element; it is also used in the medical industry. In this study, it was intended to concentrate iodide by utilizing FO system and can be reused for TFT-LCD production. Cellulose triacetate (CTA) membranes were used for all these FO experiments, and potassium iodide solution was used as the feed solution. It has been found that EDTA-2Na as draw solution at pH 8 produced high water flux and minimized salt leakage. The result also demonstrated that EDTA-2Na of concentration 0.6M could achieve the highest water flux (6.69L/m2 h). Additionally, from the recovered iodide ion from pH 3-8, the I- species was found to be more than 99%, whereas I2 was measured to be less than 1%. When potassium iodide solution was used from low to high concentration (1000 ppm to 10000 ppm), the iodide rejection was found to be than more 90%. Since, CTA membrane is negatively charged and I- is anionic in nature, so it will from electrostatic repulsion and hence there will be higher rejection. The overall performance demonstrates that recovery of concentrated iodide using FO system is a promising technology.

Keywords: draw solution, EDTA-2Na, forward osmosis, potassium iodide

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Authors: Umairah Natasya Binti Mohd Omeershffudin, Suresh Kumar

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Klebsiella pneumoniae, a Gram-negative enteric bacterium that causes nosocomial and urinary tract infections. Particular concern is the global emergence of multidrug-resistant (MDR) strains of Klebsiella pneumoniae. Characterization of antibiotic resistance determinants at the genomic level plays a critical role in understanding, and potentially controlling, the spread of multidrug-resistant (MDR) pathogens. In this study, drug-resistant Klebsiella pneumoniae strain 825795-1 was investigated with extensive computational approaches aimed at identifying novel drug targets among hypothetical proteins. We have analyzed 1099 hypothetical proteins available in genome. We have used in-silico genome subtraction methodology to design potential and pathogen-specific drug targets against Klebsiella pneumoniae. We employed bioinformatics tools to subtract the strain-specific paralogous and host-specific homologous sequences from the bacterial proteome. The sorted 645 proteins were further refined to identify the essential genes in the pathogenic bacterium using the database of essential genes (DEG). We found 135 unique essential proteins in the target proteome that could be utilized as novel targets to design newer drugs. Further, we identified 49 cytoplasmic protein as potential drug targets through sub-cellular localization prediction. Further, we investigated these proteins in the DrugBank databases, and 11 of the unique essential proteins showed druggability according to the FDA approved drug bank databases with diverse broad-spectrum property. The results of this study will facilitate discovery of new drugs against Klebsiella pneumoniae.

Keywords: pneumonia, drug target, hypothetical protein, subtractive genomics

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Authors: Hasmadi Mamat, Noorfarahzilah Masri

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The search for lesser known and underutilized crops, many of which are potentially valuable as human and animal foods has been the focus of research in recent years. Tarap (Artocarpus odoratisimmus) is one of the most delicious tropical fruit and can be found extensively in Borneo, particularly in Sabah and Sarawak. This study was conducted in order to determine the proximate composition, mineral contents as well as to study the effect of the seed flour on the quality of bread produced. Tarap seed powder (TSP) was incorporated (up to 20%) with wheat flour and used to produce bread. The moisture content, ash, protein, fat, ash, carbohydrates, and dietary fiber were measured using AOAC methods while the mineral content was determined using AAS. The effect of substitution of wheat flour with Tarap seed flour on the quality of dough and bread was investigated using various techniques. Farinograph tests were applied to determine the effect of seaweed powder on the rheological properties of wheat flour dough, while texture profile analysis (TPA) was used to measure the textural properties of the final product. Besides that sensory evaluations were also conducted. On a dry weight basis, the TSP was composed of 12.50% moisture, 8.78% protein, 15.60% fat, 1.17% ash, 49.65% carbohydrate and 12.30% of crude fiber. The highest mineral found were Mg, followed by K, Ca, Fe and Na respectively. Farinograh results found that as TSP percentage increased, dough consistency, water absorption capacity and development time of dough decreased. Sensory analysis results showed that bread with 10% of TSP was the most accepted by panelists where the highest acceptability score were found for aroma, taste, colour, crumb texture as well as overall acceptance. The breads with more than 10% of TSP obtained lower acceptability score in most of attributes tested.

Keywords: tarap seed, proximate analysis, bread, sensory evaluation

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Authors: M. Soltani, K. Pakzad, A. Haghigh-Khiyabani, M. Alavi, R. Naderi, M. Castex

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Effect of probiotic Bactocell (Pediococcus acidilactici) as a supplementary diet was studied on post-larvae 12-15 of white leg shrimp (Litopenaeus vannamei) (150000 PL/0.5 h pond, average body weight=0.02 g) growth performance under farm condition for 102 days at water quality parameters consisting of temperature at 30.5-36οC, dissolved oxygen 4.1-6.6 mg/l, salinity 40-54 g/l, turbidity 35-110 cm, ammonia 0.1-0.8 mg/l and nitrite 0.1-0.9 mg/l. Also, the resistance level of the treated shrimps was assessed against a virulent strain of Vibrio paraheamolyticus as intramuscular injection route at 1.4 x 106 cells/shrimp. Significantly higher growth rate (11.3±1.54 g) and lower feed conversion ratio (1.1) were obtained in shrimps fed diets supplemented with Bactocell at 350 g/ tone feed compared to other treatments of 250 g Bactocell/ton feed (10.8±2 g, 1.3), 500 g Bactocell/ton feed (10.3±1.7 g, 1.3) and untreated control (10.1±2 g, 1.4). Also, thermal growth coefficient (0.057%) and protein efficiency ratio (2.13) were significantly improved in shrimps fed diets supplemented with Bactocell at 350 g/ton feed compare to other groups. Shrimps fed diet supplemented with Bactocell at 350 g/tone feed showed significantly higher protein content (72.56%) in their carcass composition than treatments of 250 g/ton feed (65.9%), 500 g/ton feed (67.5%) and control group (65.9%), while the carcass contents of moisture, lipid and ash in all shrimp groups were not significantly affected by different concentrations of Bactocell. No mortality occurred in the experimentally infected shrimps fed with Bactocell at 500 g/tone feed after 7 hours post-challenge with V. parahemolyticus. The mortality levels of 100%, 40%, 50% and 70% were obtained in shrimps fed with 0.0, 500 g/tone feed, 350 g/ton feed and 250 g/ton feed, respectively 14 hours post-infection. Also, the cumulative mortalities were achieved in 100%, 92% and 81% in shrimps few with Bactocell at 500 g/ton feed, 250 g/ton feed and 350 g/ton feed, respectively.

Keywords: litopenaeus vannamei, vibrio paraheamolyticus, pediococcus acidilactici, growth performance, bactocell

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Authors: V. Ravindra, P. A. Saikiran, M. Ramgopal

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This paper presents an energy analysis of a solar assisted trigeneration system using an Ejector for dairy applications. The working fluid in the trigeneration loop is Supercritical CO₂. The trigeneration system is a combination of Brayton cycle and ejector based vapor compression refrigeration cycle. The heating and cooling outputs are used for simultaneous pasteurization and chilling of the milk. The electrical power is used to drive the auxiliary equipment in the dairy plant. A numerical simulation is done with Engineering Equation Solver (EES), and a parametric analysis is performed by varying the operating variables over a meaningful range. The results show that the overall performance index decreases with increase in ambient temperature. For an ejector based system, the compressor work and cooling output are significant output quantities. An increase in total mass flow rate of the refrigerant (primary + secondary) results in an increase in the compressor work and cooling output.

Keywords: trigeneration, solar thermal, supercritical CO₂, ejector

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Authors: Maryam Ehsanpou, Majid Afkhami, Flora Mohammadizadeh, Amirhoushang Bahri, Rastin Afkhami

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Within the frame of a biodiversity and bioactivity study of marine macro organisms from the Persian Gulf, three hard coral species extracts were investigated for cytotoxic, antibacterial and antifungal activities against five human pathogenic microorganisms. All concentrations of extracts from three hard corals showed no antifungal activity towards the tested strains. In antibacterial assays, the hard coral extracts showed significant activity solely against Staphylococcus aureus with MICs ranging from 3 to 9 μg/ml. The highest antibacterial activity was found in the aqueous methanol extract of Porites compressa with an inhibition zone of 22 mm against Staphylococcus aureus at 18 μg/ml extract concentration. Methanol extracts from Porites harrisoi and Porites compressa exhibited only weak cytotoxic activities. It is important for future research to concentrate on finding the mechanisms employed by corals to defend themselves against invasion, the mechanism of infections and the type of chemical compounds in coral extracts that inhibit antibacterial growth or proliferation in underexplored areas such as the Persian Gulf.

Keywords: antibacterial, antifungal, cytotoxic, hard corals, Persian Gulf

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Authors: Moustafa Elhamouly, Masayuki Shimada

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The production of competent oocytes is essential for reproductivity in mammals. Maintenance of mitochondrial efficiency is required to supply the ATP necessary for granulosa cell proliferation during the follicular development process. Treatment with Pyrroloquinoline quinone (PQQ) has been reported to increase the number of ovulated oocytes and pups per delivery in mice by maintaining healthy mitochondrial function. This study aimed to elucidate how PQQ maintains mitochondrial function during ovarian follicle growth. To do this, both in vitro and in vivo experiments were performed with granulosa cells from superovulated immature (3-week-old) mice that were pretreated with or without PQQ. The effects of PQQ on beta-oxidation, mitochondrial function, mitophagy, and mitochondrial biogenesis were examined. PQQ increased beta-oxidation-related genes and CPT1 protein content in granulosa cells and this was associated with a decreased phosphorylation of P38 signaling protein. Using the fatty acid oxidation assay on the flux analyzer, PQQ increased the reliance of beta-oxidation on the endogenous fatty acids and was associated with a mild UCP-dependant mitochondrial uncoupling, ATP production, mitophagy, and mitochondrial biogenesis. PQQ also increased the expression of endogenous antioxidant enzymes. Thus, PQQ induced beta-oxidation in growing granulosa cells relying on endogenous fatty acids. And reduced the Reactive oxygen species (ROS) production by inducing a mild mitochondrial uncoupling with keeping high mitochondrial function. Damaged mitochondria were recycled by the induced mitophagy and replaced by the increased mitochondrial biogenesis. Collectively, PQQ may enhance reproductivity by maintaining the efficiency of mitochondria to produce enough ATP required for normal folliculogenesis.

Keywords: granulosa cells, mitochondrial uncoupling, mitophagy, pyrroloquinoline quinone (PQQ), reactive oxygen species (ROS).

Procedia PDF Downloads 76