Search results for: milk proteins

Authors: Carolina Gutierrez-Cortes, Hector Suarez, Gustavo Buitrago

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Bacteriocins are antimicrobial peptides produced by bacteria as a competitive strategy for substrate and habitat. Those peptides have a potential use as food biopreservatives due to their antimicrobial activity against foodborne pathogens, avoiding the use of additives that can be harmful to consumers. The industrial production of bacteriocins is currently expensive; one of the options to be competitive is the development of economic culture media, for example, with the use of agro-industrial wastes such as whey. This study evaluated the growth and production of bacteriocins from four strains: Pediococcus pentosaceus 63, Pediococcus pentosaceus 145, Pediococcus pentosaceus 146 and Pediococcus pentosaceus 147 isolated from ‘minas cheese’ (artisanal cheese made from raw milk in the state of Minas Gerais, Brazil) in order to select a strain with growth at high rates and higher antimicrobial activity against Listeria monocytogenes 104 after incubation on the culture medium designed with whey and other components. The media used were: MRS broth, modified MRS broth (using different sources of carbon and nitrogen and different amounts of micronutrients) and a culture medium designed by a factorial design using whey and other components. The final biomass concentrations of the four strains in MRS broth after 24 hours of incubation were very similar 9.25, 9.33, 9.25 and 9.22 (log CFU/mL) for P. pentosaceus 63, P. pentosaceus 145, P. pentosaceus 146 and P. pentosaceus 147 respectively. In the same assays, antimicrobial activity of 3200 AU/mL for the first three and of 12800 AU/mL for P. pentosaceus 147 were obtained. Culture of P. pentosaceus 63 on modified MRS broth, showed the effect of some sources of carbon on the activity of bacteriocin, obtaining 12800 AU/mL with dextrose and 25600 AU/mL with maltose. Cultures of P. pentosaceus 145, 146 and 147 with these same sugars presented activity of 12800 AU/mL. It was observed that the modified MRS medium using whey increased the antimicrobial activity of the strains at 16000, 6400, 16000 and 19200 AU/mL for each strain respectively, keeping the biomass at values close to 9 log units. About nitrogen sources, it was observed that the combination of peptone (10 g /L), meat extract (10 g/L) and yeast extract (5 g/L) promoted the highest activity (12800 AU/mL), and in all cases MgSO4, MnSO4, K2HPO4 and ammonium citrate at low concentrations adversely affected bacteriocin production. Because P. pentosaceus 147 showed the highest antimicrobial activity in the presence of whey, it was used to evaluate the culture medium (peptone (10 g/L), meat extract (8 g/L), yeast extract (2 g/L), Tween® 80 (1 g/L), ammonium citrate (2 g/L), sodium acetate (5 g/L), MgSO4 (0.2 g/L), MnSO4 (0.04 g/L)). With the designed medium added with whey, 9.34 log units of biomass concentration and 19200 AU/mL were achieved for P. pentosaceus 147. The above suggest that the new medium promotes the antimicrobial activity of P. pentosaceus 147 allowing the use of an economic medium using whey.

Keywords: antimicrobial activity, bacteriocins, pediococcus, whey

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Authors: Mohammed Alasel, Michael Keusgen

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Gold is a noble metal; in its nano-scale level (e.g. spherical nanoparticles), the conduction electrons are triggered to collectively oscillate with a resonant frequency when certain wavelengths of electromagnetic radiation interact with its surface; this phenomenon is known as surface plasmon resonance (SPR). SPR is responsible for giving the gold nanoparticles its intense red color depending mainly on its size, shape and distance between nanoparticles. A decreased distance between gold nanoparticles results in aggregation of them causing a change in color from red to blue. This aggregation enables gold nanoparticles to serve as a sensitive biosensoric indicator. In the proposed work, gold nanoparticles were modified with two proteins: i) Borrelia antigen, variable lipoprotein surface-exposed protein (VlsE), and ii) protein A. VlsE antigen induces a strong antibody response against Lyme disease and can be detected from early to late phase during the disease in humans infected with Borrelia. In addition, it shows low cross-reaction with the other non-pathogenic Borrelia strains. The high specificity of VlsE antigen to anti-Borrelia antibodies, combined simultaneously with the high specificity of protein A to the Fc region of all IgG human antibodies, was utilized to develop a rapid test for serological point of care diagnosis of borreliosis in human serum. Only in the presence of anti-Borrelia antibodies in the serum probe, an aggregation of gold nanoparticles can be observed, which is visible by a concentration-dependent colour shift from red (low IgG) to blue (high IgG). Experiments showed it is clearly possible to distinguish between positive and negative sera samples using a simple suspension of the two-protein modified gold nanoparticles in a very short time (30 minutes). The proposed work showed the potential of using such modified gold nanoparticles generally for serological diagnosis. Improved specificity and reduced assay time can be archived in applying increased salt concentrations combined with decreased pH values (pH 5).

Keywords: gold nanoparticles, gold aggregation, serological diagnosis, protein A, lyme borreliosis

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Authors: Qi LI, Xu Lin, Nengming Lin

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Objective: The aim of this study was to investigate the role of desmocollin 2 (DSC2) protein in the proliferation, migration and drug resistance of lung cancer cells. Method: CCK-8 assays and colony formation assays were used to evaluate the effect of dsc2 regulation on cancer cell viability and colony formation. Transwell assays and wound healing assays were also performed. Cell flow double staining was used to detect the apoptosis rate of cells with DSC2, which was added cisplatin. Western blot assay was used to detect cell cycle, PI3k/Akt and apoptosis-related proteins. Results: Our data showed that dsc2 is upregulated in clinical lung cancer tissues compared with pericarcinomatous tissues, and it is differentially expressed in lung cancer cell lines. The down-regulation of dsc2 in A549 and H358 lung cancer cells significantly suppressed the cell proliferation, metastasis, and motility. In contrast, the opposite effects were observed in overexpression of dsc2 both in H23 and PC9 cell lines. In addition to lung adenocarcinoma cell lines, we also examined its expression in lung squamous cell lines, such as H226. Western blotting showed that dsc2 could reduce the level of phosphorylated Akt (Ser 473) and p-mTOR. Thus, it is speculated that dsc2 up-regulation promotes proliferation and invasiveness through activation of the PI3K/AKT pathway. Also, knockdown of dsc2 in A549 and H226 could significantly decreased in the levels of cyclinB and wee1 protein. Additionally, flow cytometry showed that dsc2 knockdown combined with cisplatin could significantly enhance cell apoptosis rate. Conclusion: These data suggest that dsc2 promotes the proliferation and migration of lung cancer cells in vitro. Also, the results suggested that dsc2 could affect the cell cycle and apoptosis of lung cells. Furthermore, knockdown of dsc2 could sensitize cisplatin in both lung adenocarcinoma and lung squamous cell lines. Thus we suggested that dsc2 can be used as a therapeutic target for lung cancer.

Keywords: desmocollin 2, cisplatin, lung cancer, PI3K/AKT, lung squamous cell

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Authors: Nishanthi N. S., Srikanth Vedantam

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Intracellular delivery of materials has always proved to be a challenge in research and therapeutic applications. Usually, vector-based methods, such as liposomes and polymeric materials, and physical methods, such as electroporation and sonoporation have been used for introducing nucleic acids or proteins. Reliance on exogenous materials, toxicity, off-target effects was the short-comings of these methods. Microinjection was an alternative process which addressed the above drawbacks. However, its low throughput had hindered its adoption widely. Mechanical deformation of cells by squeezing them through constriction channel can cause the temporary development of pores that would facilitate non-targeted diffusion of materials. Advantages of this method include high efficiency in intracellular delivery, a wide choice of materials, improved viability and high throughput. This cell squeezing process can be studied deeper by employing simple models and efficient computational procedures. In our current work, we present a finite sized dissipative particle dynamics (FDPD) model to simulate the dynamics of the cell flowing through a constricted channel. The cell is modeled as a capsule with FDPD particles connected through a spring network to represent the membrane. The total energy of the capsule is associated with linear and radial springs in addition to constraint of the fixed area. By performing detailed simulations, we studied the strain on the membrane of the capsule for channels with varying constriction heights. The strain on the capsule membrane was found to be similar though the constriction heights vary. When strain on the membrane was correlated to the development of pores, we found higher porosity in capsule flowing in wider channel. This is due to localization of strain to a smaller region in the narrow constriction channel. But the residence time of the capsule increased as the channel constriction narrowed indicating that strain for an increased time will cause less cell viability.

Keywords: capsule, cell squeezing, dissipative particle dynamics, intracellular delivery, microfluidics, numerical simulations

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Authors: G. Disciglio, G. Gatta, F. Lops, A. Libutti, A. Tarantino, E. Tarantino

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The experimental trial was carried out in open field at Foggia district (Apulia Region, Southern Italy), during the spring-summer season 2014, in order to evaluate the effect of four biostimulant products (RadiconÒ, Viormon plusÒ, LysodinÒ and SiaptonÒ 10L), compared with a control (no biostimulant), on the infestation of processing tomato crop (cv Dres) by the chlorophyll-lacking root parasite Phelipanche ramosa. Biostimulants consist in different categories of products (microbial inoculants, humic and fulvic acids, hydrolyzed proteins and aminoacids, seaweed extracts) which play various roles in plant growing, including the improvement of crop resistance and quali-quantitative characteristics of yield. The experimental trial was arranged according to a complete randomized block design with five treatments, each of one replicated three times. The processing tomato seedlings were transplanted on 5 May 2014. Throughout the crop cycle, P. ramosa infestation was assessed according to the number of emerged shoots (branched plants) counted in each plot, at 66, 78 and 92 day after transplanting. The tomato fruits were harvested at full-stage of maturity on 8 August 2014. From each plot, the marketable yield was measured and the quali-quantitative yield parameters (mean weight, dry matter content, colour coordinate, colour index and soluble solids content of the fruits) were determined. The whole dataset was tested according to the basic assumptions for the analysis of variance (ANOVA) and the differences between the means were determined using Tukey’s tests at the 5% probability level. The results of the study showed that none of the applied biostimulants provided a whole control of Phelipanche, although some positive effects were obtained from their application. To this respect, the RadiconÒ appeared to be the most effective in reducing the infestation of this root-parasite in tomato crop. This treatment also gave the higher tomato yield.

Keywords: biostimulant, control methods, Phelipanche ramosa, tomato crop

Procedia PDF Downloads 279

Authors: Jelka Pleadin, Greta Krešić, Tina Lešić, Ana Vulić, Renata Barić, Tanja Bogdanović, Dražen Oraić, Ana Legac, Snježana Zrnčić

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Fish has been acknowledged as an integral component of a well-balanced diet, providing a healthy source of energy, high-quality proteins, vitamins, essential minerals and, especially, n-3 long-chain polyunsaturated fatty acids (n-3 LC PUFA), mainly eicosapentaenoic acid (20:5 n-3 EPA), and docosahexaenoicacid, (22:6 n-3 DHA), whose pleiotropic effects in terms of health promotion and disease prevention have been increasingly recognised. In this study, the fatty acids composition of three commercially important farmed fish species: sea bream (Sparus aurata), sea bass (Dicentrarchus labrax) and dentex (Dentex dentex) was investigated. In total, 60 fish samples were retrieved during 2015 (n = 30) and 2016 (n = 30) from different locations in the Adriatic Sea. Methyl esters of fatty acids were analysed using gas chromatography (GC) with flame ionization detection (FID). The results show that the most represented fatty acid in all three analysed species is oleic acid (C18:1n-9, OA), followed by linoleic acid (C18:2n-6, LA) and palmitic acid (C16:0, PA). Dentex was shown to have two to four times higher eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid content as compared to sea bream and sea bass. The recommended n-6/n-3 ratio was determined in all fish species but obtained results pointed to statistically significant differences (p < 0.05) in fatty acid composition among the analysed fish species and their potential as a dietary source of valuable fatty acids. Sea bass and sea bream had a significantly higher proportion of n-6 fatty acids, while dentex had a significantly higher proportion of n-3 (C18:4n-3, C20:4n-3, EPA, DHA) fatty acids. A higher hypocholesterolaemic and hypercholesterolaemic fatty acids (HH) ratio was determined for sea bass and sea bream, which comes as the consequence of a lower share of SFA determined in these two species in comparison to dentex. Since the analysed fish species vary in their fatty acids composition consumption of diverse fish species would be advisable. Based on the established lipid quality indicators, dentex, a fish species underutilised by the aquaculture, seems to be a highly recommendable and important source of fatty acids recommended to be included into the human diet.

Keywords: dentex, fatty acids, farmed fish, sea bass, sea bream

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Authors: Eitan Yefenof

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Glucocorticoid (GC) hormones, e.g. Dexamethasone and Prednisone, are widely used in the therapy of leukemia and lymphoma owing to their apoptogenic effect on lymphoid cells. However, the emergence of GC resistant cells during therapy is a major cause for treatment failure, urging the need for novel strategies that maintain leukemia sensitivity to the pro-apoptotic activity of GCs. GCs act by binding to the GC receptor (GR), which, in its inactive state, is sequestered in the cytosol by a multi-subunit complex of heat shock proteins. Upon ligand binding, the complex dissociates, allowing GR activation and translocation to the nucleus, where it regulates transcription of multiple genes. We demonstrated that in addition to gene expression, GR also regulates microRNA (miR) expression. Deep-sequencing analysis revealed 14 miRs that are regulated in GC-sensitive but resistant leukemias upon treatment with GC. GC up-regulates miR-103, miR-15~16 and miR-30e/d, while down-regulates miR-17, mir-18a, miR-19a, miR-19b, miR-20a and miR-92a (members of the miR-17∼92a multi-cistron). Upon transfection, miR-103 confers GC apoptotic sensitivity to otherwise GC-resistant cell. Furthermore, knocking down miR-103 expression reduces the GC apoptotic response of sensitive cells. miR-103 abrogates c-Myc expression, an oncogenic transcription factor which is deregulated in many cancers. In addition, miR-103 up-regulates Bim, a pro-apoptotic protein crucial for GC-induced death. Activated glycogen synthase kinase 3 (GSK3) is also crucial for GC-induced apoptosis. GSK3 is active in GC-sensitive but not in GC-resistant cells. We found that GSK3 associates with the GR multi-subunit complex. Upon GC exposure, it dissociates from the GR and interacts with Bim to enable activation of the mitochondrial apoptosis pathway. miR-103 mediated c-Myc ablation is followed by down-regulation of the multi-cistron miR-17~92a, in particular miR-18a and miR-20a. miR-18a targets GR for degradation whereas miR-20a targets Bim degradation. Hence, miR-103 acts, in concert with Bim and GR, as a "tumor suppressor" that leads to reduced proliferation, cell-cycle arrest and cell death. We suggest that miR-103 can provide a diagnostic tool that predicts the sensitivity of leukemia to GC based therapy. Furthermore, exosomal delivery of miR-103 or up-regulation of the endogenous miR-103 could confer apoptotic sensitivity to resistant cells at the outset, thus becoming a useful therapeutic tool combined with GCs.

Keywords: apoptosis, leukemia, micro-RNA, steroids

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Authors: Min-Chien Su, Tzu-Hsuan Hsu, Guan-Xuan Wu, Shyh-Ming Kuo

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Lung cancer is one of the clinically challenging malignant diseases worldwide. For efficient therapeutics in cancer, combination therapy has developed to acquire a better outcome. PLK-1 was one of the major factors affecting cell mitosis in cancer cells, its inhibitor Bi6727 was proven effective in treating several different cancers namely oral cancer, colon cancer and lung cancer. Despite its low toxicity toward normal cells compared to traditional chemotherapy, it is still yet to be evaluated in detail. Livistona Chinensis (LC) is a Chinese herb that used as a traditional prescription to treat lung cancer. Due to the uncertainty of the efficacy of LC, we utilized a water extraction method to extract the Livistona Chinensis and then lyophilized into powder for further study. In this study we investigated the antiproliferation activities of Bi6727 and LC extracts (LCE) on A549 non-small lung cancer cells. The IC50 of Bi6727 and LCE on A549 are 60 nM and 0.8 mg/mL, respectively. The fluorescent staining images shown nucleolus damage in cells treated with Bi6727 and mitochondrial damage after treated with LCE. A549 cells treated with Bi6727 and LCE showed increased expression of Bax, Caspase-3 and Caspase-9 proteins from Western blot assay. LCE also inhibited A549 cells growth keeping cells at G2-M phase from cell cycle assay. Apoptosis assay results showed that LCE induced late apoptosis of A549 cells. JC-1 assay showed that the mitochondria damaged at the LCE concentration of 0.4 mg/mL. In our preliminary anti-proliferation test of combined LCE and Bi-6727 on A549 cells, we found a dramatically decrease in proliferation after treated with LCE first for 24-h and then Bi-6727 for extra 24-h. This was an important finding regarding synergistic anti-proliferation effect of these drugs, However, the usage, the application sequence of LCE and Bi-6727 on A549 cells and their related mechanisms still need to be evaluated. In summary, the drugs exerted anti-proliferation effect on A549 cells independently. We hopefully combine the usage of these two drugs will bring a different and potential outcome in treating lung cancer.

Keywords: anti-proliferation, A549, Livistona Chinensis fruit extracts, PLK-1 inhibitor

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Authors: G. S. Sumanasekara, A. Balasuriya

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Introduction: In Sri Lanka studies reveal higher trend in prevalence of diabetes. The office employees have sedentary life style and their eating patterns changed due to nutritional transition. Further overall, urban and rural pre diabetes is also increasing. Objectives - Study the general food pattern of office employees and its relation to overweight/obesity and prevalence of diabetes among them. Method: The data was collected from office employees between 30-60 years (n-400).Data analyzed using SPSS 16 version.The Study design was a descriptive cross sectional study. The study setting was Ministry of Economic Development. Anthropometric measurements and blood glucose assessed by trained nurses. Dietary pattern was studied through a food frequency questionairre thereby calculated daily nutrient intakes. Results: Mean age of office employees were 38.98 SD (7.033) CI=95%) and 245 females (61.2%) 155 males (38.8 %) ,Nationality includes Sinhala (67.5%), Tamil(20%), and Muslims (12.5%).Owerweight(7,1.8%), obese male(36,9%), obese female(66,16%)/ diabetes/obese(18,4.5%) out of 127(31.8%) who were above the normal BMI whereas 273(68.2) were within the normal. Mean BMI was 24.1593.Mean Blood sugar level was 104.646,SD(16.018).12% consume tobacco products,17.8 consumed alcohol.15.8% had nutrition training. Two main dietary patterns identified who were vegetarians and non vegetarians .Mean energy intake 1727.1, (SD 4.97), Mean protein consumption(11.33, SD 1.811), Mean fat consumption(24.07, SD 4.131),Mean CHO consumption (64.56, SD 4.54), Mean Fibre (30.05, SD 17.9), Mean cholesterol(16.85, SD 17.22), Energy intake was higher in non vegetarians and larger propotion of energy derived from proteins , and fat. Their carbohydrate and cholesterol intake was also higher. Tamils were mostly vegetarians. Mainly BMI were within normal range(18.5-23.5) whereas Muslims who had higher energy intakes showed BMI above the normal. Conclusion: Two distinct dietary patterns identified. Different ethnic groups consume different diets with different nutrient composition. Dietary pattern has a relation to overweight. Overweight related to high blood glucose levels but some overweight subjects do not show any relation.

Keywords: obesity, overweight, diabetes, dietary pattern, nutrition, BMI, non communicable disease

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Authors: Sadaf Moeez, Madiha Khalid, Zoya Khalid, Sania Shaheen, Sumbul Khalid

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Background: Inter-individual variation in response to metformin, which has been considered as a first line therapy for T2DM treatment is considerable. In the current study, it was aimed to investigate the impact of two genetic variants Leu125Phe (rs77474263) and Gly64Asp (rs77630697) in gene SLC47A1 on the clinical efficacy of metformin in T2DM Pakistani patients. Methods: The study included 800 T2DM patients (400 metformin responders and 400 metformin non-responders) along with 400 ethnically matched healthy individuals. The genotypes were determined by allele-specific polymerase chain reaction. In-silico analysis was done to confirm the effect of the two SNPs on the structure of genes. Association was statistically determined using SPSS software. Results: Minor allele frequency for rs77474263 and rs77630697 was 0.13 and 0.12. For SLC47A1 rs77474263 the homozygotes of one mutant allele ‘T’ (CT) of rs77474263 variant were fewer in metformin responders than metformin non-responders (29.2% vs. 35.5 %). Likewise, the efficacy was further reduced (7.2% vs. 4.0 %) in homozygotes of two copies of ‘T’ allele (TT). Remarkably, T2DM cases with two copies of allele ‘C’ (CC) had 2.11 times more probability to respond towards metformin monotherapy. For SLC47A1 rs77630697 the homozygotes of one mutant allele ‘A’ (GA) of rs77630697 variant were fewer in metformin responders than metformin non-responders (33.5% vs. 43.0 %). Likewise, the efficacy was further reduced (8.5% vs. 4.5%) in homozygotes of two copies of ‘A’ allele (AA). Remarkably, T2DM cases with two copies of allele ‘G’ (GG) had 2.41 times more probability to respond towards metformin monotherapy. In-silico analysis revealed that these two variants affect the structure and stability of their corresponding proteins. Conclusion: The present data suggest that SLC47A1 Leu125Phe (rs77474263) and Gly64Asp (rs77630697) polymorphisms were associated with the therapeutic response of metformin in T2DM patients of Pakistan.

Keywords: diabetes, T2DM, SLC47A1, Pakistan, polymorphism

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Authors: Vasudha Devi, Arul Amuthan, K. Narayanan, Praveen KS, Venkata Rao J

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Background: Siddha Medicine is one of the Indian traditional medical systems, in which the cancer disease is mentioned as 'putrunoi' which literally means the disease of growth like termite mound. There are number of formulations available for the treatment of cancer disease. Neeradi muthu vallathymezugu (NMV) and thamira kattu chendooram (TKC) are two drugs commonly prescribed by Siddha physicians. These drugs have been clinically reported to be safe and effective when given orally. Though these formulations are in practice for centuries, no efforts have been made to standardize them and explore their anti-cancer potential systematically. Objective: To compare the cytotoxic activity of NMV and TKC with doxorubicin using cancer cell lines. Materials and methods: For this study, ethanol extract of NMV was taken, whereas TKC was used as such. In vitro cytotoxic activity was evaluated by sulphorhodamine (SRB) assay against human hepatic cancer cells (HepG2), human breast cancer cells (MCF-7) and human cervical cancer cells [KeLa]. Doxorubicin was used as the standard. The SRB assay is based on the ability of cellular proteins to bind with sulphorhodamine-B. The number of live cells in drug treated cell lines directly affects the color formation in the assay, which is estimated calorimetrically by measuring the absorbance at 540 nm to calculate the cytotoxicity (inhibitory concentration - IC50 value) of the drug. Results: The IC50values of NMV, TKC and doxorubicin against HepG2 were 3.08 µg/ml, 20.21 µg/ml and 1.21µg/ml respectively. In MCF-7, it was 11.75 µg/ml, 17.67 µg/ml and 2.8µg/ml. In HeLa, the values were 24.76 µg/ml, 73.35 µg/ml and 1.12µg/ml. Conclusions: The study proves the possible anti-cancer potential of these two formulations. Compared to TKC, NMV showed good cytotoxic effect even at low dose. Human hepatic cancer cells responded well even at very low dose, when compared to other cancer cells. Though, cytotoxic potential of these compounds was found to be less compared to doxorubicin, the isolated lead compound may have the potential to be used as an anticancer drug clinically.

Keywords: Neeradi muthu vallathymezugu (Hydnocarpus laurifolia), thamira kattu chendooram, cytotoxicity, in-vitro, Siddha Medicine

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Authors: Shadia Al-Bahlani, Ruqaya Al-Rashdi, Shadia Al-Sinawi, Maya Al-Bahri

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Background: Breast cancer is the most common cancer in women worldwide. Triple-negative breast cancer (TNBC) is an aggressive type of breast cancer, which is defined by the absence of Estrogen (ER), Progesterone (PR) and Human epidermal growth factor (Her-2) receptors. The calpain system plays an important role in many cellular processes including apoptosis, necrosis, cell signaling and proliferation. The role of clapins in pathogenesis and tumor progression has been studied in certain cancer types; however, its definite role is not yet established in breast cancer especially in the TNBC subtype. Objectives: This study aims to measure calpain-1 expression and correlate this measurement with the proliferating/apoptotic index as well with the prognostic factors in TNBC patients’ tissue. Materials and Methods: Thirty nine paraffin blocks from patients diagnosed with TNBC were used to measure the expression of calpain-1 and Ki-67 (proliferating marker) proteins using immunohistochemistry. Apoptosis was assessed morphological and biochemically using conventional Haematoxylin and Eosin (H&E) staining method and terminal deoxynucleotidyl transferase-mediate dUTP nick and labeling (TUNEL) assay respectively. Data was statistically analyzed using Pearson X2 test of association. Results: Calpain-1 content was visualized in the nucleus of the TNBC cells and its expression varied from low to high among the patients tissue. Calpain expression showed no significant correlation with the proliferating/apoptotic index as well with the clinicopathological variables. Apoptotic counts quantified by H&E staining showed significant association with the apoptotic TUNEL assay, validating both approaches. Conclusion: Although calpain-1 expression showed no significant association with the clinical outcome, its variable level of expression might indicate a hidden role in breast cancer tissue. Larger number of samples and different mode of assessments are needed to fully investigate such role. Exploring the involvement of calpain-1 in cancer progression might help in considering it as a biomarker of breast cancer.

Keywords: breast cancer, calpain, apoptosis, prognosis

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Authors: P. Kiran Kumar, S. Vijaya Krishna, Kavita Verma1, V. Himabindu

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Concern about global warming and energy security has led to increased biomass utilization as an alternative feedstock to fossil fuels. Biomass is a promising feedstock since it is abundant and cheap and can be transformed into fuels and chemical products. Microalgae biofuels are likely to have a much lower impact on the environment. Microalgae cultivation using sewage with industrial flue gases is a promising concept for integrated biodiesel production, CO₂ sequestration, and nutrients recovery. Autotrophic, Mixotrophic, and Heterotrophic are the three modes of cultivation for microalgae biomass. Several mechanical and chemical processes are available for the extraction of lipids/oily components from microalgae biomass. In organic solvent extraction methods, a prior drying of biomass and recovery of the solvent is required, which are energy-intensive. Thus, the hydrothermal process overcomes the drawbacks of conventional solvent extraction methods. In the hydrothermal process, the biomass is converted into oily components by processing in a hot, pressurized water environment. In this process, in addition to the lipid fraction of microalgae, other value-added products such as proteins, carbohydrates, and nutrients can also be recovered. In the present study was (Scenedesmus quadricauda) was isolated and cultivated in autotrophic, heterotrophic, and mixotrophically using sewage wastewater and industrial flue gas in batch and continuous mode. The harvested algae biomass from S. quadricauda was used for the recovery of lipids and bio-oil. The lipids were extracted from the algal biomass using sonication as a cell disruption method followed by solvent (Hexane) extraction, and the lipid yield obtained was 8.3 wt% with Palmitic acid, Oleic acid, and Octadeonoic acid as fatty acids. The hydrothermal process was also carried out for extraction of bio-oil, and the yield obtained was 18wt%. The bio-oil compounds such as nitrogenous compounds, organic acids, and esters, phenolics, hydrocarbons, and alkanes were obtained by the hydrothermal process of algal biomass. Nutrients such as NO₃⁻ (68%) and PO₄⁻ (15%) were also recovered along with bio-oil in the hydrothermal process.

Keywords: flue gas, hydrothermal process, microalgae, sewage wastewater, sonication

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Authors: C. Flynn, Q. Yuan, C. Reinhardt

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Alzheimer’s Disease (AD) is a progressive neurodegenerative disorder affecting broad areas of the cerebral cortex and hippocampus. More than 95% of AD cases are representative of sporadic AD, where both genetic and environmental risk factors play a role. The main pathological features of AD include the widespread deposition of amyloid-beta and neurofibrillary tau tangles in the brain. The earliest brain pathology related to AD has been defined as hyperphosphorylated soluble tau in the noradrenergic locus coeruleus (LC) neurons, characterized by Braak. However, the cause of this pathology and the ultimate progression of AD is not understood. Increasing research points to a connection between the gut microbiota and the brain, and mounting evidence has shown that there is a bidirectional interaction between the two, known as the gut-brain axis. This axis can allow for bidirectional movement of neuroinflammatory cytokines and pathogenic misfolded proteins, as seen in AD. Prebiotics and probiotics have been shown to have a beneficial effect on gut health and can strengthen the gut-barrier as well as the blood-brain barrier, preventing the spread of these pathogens across the gut-brain axis. Our laboratory has recently established a pretangle tau rat model, in which we selectively express pseudo-phosphorylated human tau (htauE14) in the LC neurons of TH-Cre rats. LC htauE14 produced pathological changes in rats resembling those of the preclinical AD pathology (reduced olfactory discrimination and LC degeneration). In this work, we will investigate the effects of pre/probiotic ingestion on AD behavioral deficits, blood inflammation/cytokines, and various brain markers in our experimental rat model of AD. Rats will be infused with an adeno-associated viral vector containing a human tau gene pseudophosphorylated at 14 sites (common in LC pretangles) into 2-3 month TH-Cre rats. Fecal and blood samples will be taken at pre-surgery, and various post-surgery time points. A collection of behavioral tests will be performed, and immunohistochemistry/western blotting techniques will be used to observe various biomarkers. This work aims to elucidate the relationship between gut health and AD progression by strengthening gut-brain relationship and aims to observe the overall effect on tau formation and tau pathology in AD brains.

Keywords: alzheimer’s disease, aging, gut microbiome, neurodegeneration

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Authors: Sengly Sroy, Elodie Arnaud, Adrien Servent, Sokneang In, Sylvie Avallone

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In Cambodia, fish plays an important role for local community in term of food habits, preference and contribution to several nutritional intakes. Consumed on a daily basis, fishes and their derivatives products are good sources of proteins, essential fatty acids and fat-soluble vitamins. They mainly obtain from the Tonle Sap Lake but, during the last decade, the fish population decreased drastically due to climate change and human activities as well. Contamination by agricultural residues and heavy metals were identified. However, fishes are currently used in several nutrition programs for children and pregnant women to improve their nutritional status. The aim of our work was to characterize the nutritional profile and contamination of 10 fish species consumed near the Tonle Sap Lake with a special attention to fatty acid and fat-soluble vitamin profiles. Fish samples were analyzed for their nutritional profiles (AOAC methods for macronutrients and micronutrients), their lipid content (Folch modified method), their Fatty acid (FAME method), their vitamin A (HPLC) and their heavy metals (ICP-MS). The total lipid contents ranged from 1.43 to 10.00% according to fish species. Lipid profile was mainly dominated by saturated fat (from 47.95 to 57.32%) but some fish species were particularly rich in ω-3 and ω-6 especially eicosapentaenoic acid EPA (3.05%) and docosahexaenoic acid DHA (2.82%). The more the fishes were fats, the more they contained vitamin A, DHA and EPA. Vitamin A is particularly abundant in small fishes (250.10 μg RE/100 g) compare to big ones (13.77 μg RE/100 g) because they are consumed as a whole with their organs (liver) and head. However, the contents of heavy metal in some species are higher than the maximum permitted level (MPL) from codex alimentarius, especially Mn. The results obtained provided important information on the most interesting fish in term of human nutrition and the potential risk of contaminants. The fatty acids are important for child development and pregnant women. These data are useful for supply chain stakeholders and the people in charge of nutrition program.

Keywords: fat-soluble vitamin, fatty acid, freshwater fish, lipid content, Tonle Sap Lake

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Authors: Muhammad Awais Afzal, Lorena Tuchscherr De Hauschopp, Christian Hübner

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Introduction: Autophagy is an evolutionarily conserved lysosome-dependent degradation pathway, which can be induced by extrinsic and intrinsic stressors in living systems to adapt to fluctuating environmental conditions. In the context of inflammatory stress, autophagy contributes to the elimination of invading pathogens, the regulation of innate and adaptive immune mechanisms, and regulation of inflammasome activity as well as tissue damage repair. Lysosomes can be recycled from autolysosomes by the process of autophagic lysosome reformation (ALR), which depends on the presence of several proteins including Spatacsin. Thus ALR contributes to the replenishment of lysosomes that are available for fusion with autophagosomes in situations of increased autophagic turnover, e.g., during bacterial infections, inflammatory stress or sepsis. Objectives: We aimed to assess whether ALR plays a role for cell survival in an in-vitro bacterial infection model. Methods: Mouse embryonic fibroblasts (MEFs) were isolated from wild-type mice and Spatacsin (Spg11-/-) knockout mice. Wild-type MEFs and Spg11-/- MEFs were infected with Staphylococcus aureus (multiplication of infection (MOI) used was 10). After 8 and 16 hours of infection, cell viability was assessed on BD flow cytometer through propidium iodide intake. Bacterial intake by cells was also calculated by plating cell lysates on blood agar plates. Results: in-vitro infection of MEFs with Staphylococcus aureus showed a marked decrease of cell viability in ALR deficient Spatacsin knockout (Spg11-/-) MEFs after 16 hours of infection as compared to wild-type MEFs (n=3 independent experiments; p < 0.0001) although no difference was observed for bacterial intake by both genotypes. Conclusion: Suggesting that ALR is important for the defense of invading pathogens e.g. S. aureus, we observed a marked increase of cell death in an in-vitro infection model in cells with compromised ALR.

Keywords: autophagy, autophagic lysosome reformation, bacterial infections, Staphylococcus aureus

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Authors: Maciej Sobczak, Julita Pietrzak, Tomasz Płoszaj, Agnieszka Robaszkiewicz

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Brg1, a member of SWI/SNF complex, plays a role in chromatin remodeling, therefore, regulates expression of many genes. Brg1 is an ATPase of SWI/SNF complex, thus its activity requires ATP. Through its bromodomain recognizes acetylated histone residues and evicts them, thus promoting transcriptionally active state of chromatin. One of the enzymes that is responsible for acetylation of histone residues is Ep300. It was previously shown in the literature that cooperation of Brg1 and Ep300 occurs at the promoter regions that have binding sites for E2F-family transcription factors as well as CpG islands. According to literature, approximately 20% of human cancer possess mutation in Brg1 or any other crucial SWI/SNF subunit. That phenomenon makes Brg1-Ep300 a very promising target for anti-cancer therapy. Therefore in our study, we investigated if physical interaction between Brg1 and Ep300 exists and what impact those two proteins have on key for breast cancer cells processes such as DNA damage repair and cell proliferation. Bioinformatical analysis pointed out, that genes involved in cell proliferation and DNA damage repair are overexpressed in MCF7 and MDA-MB-231 cells. Moreover, promoter regions of these genes are highly acetylated, which suggests high transcriptional activity of those sites. Notably, many of those gene possess within their promoters an E2F, Brg1 motives, as well as CpG islands and acetylated histones. Our data show that Brg1 physically interacts with Ep300, and together they regulate expression of genes involved in DNA damage repair and cell proliferation. Upon inhibiting Brg1 or Ep300, expression of vital for cancer cell survival genes such as CDK2/4, BRCA1/2, PCNA, and XRCC1 is decreased in MDA-MB-231 and MCF7 cells. Moreover, inhibition or silencing of either Brg1 or Ep300 leads to cell cycle arrest in G1. After inhibition of BRG1 or Ep300 on tested gene promoters, the repressor complex including Rb, HDAC1, and EZH2 is formed, which inhibits gene expression. These results highlight potentially significant target for targeted anticancer therapy to be introduced as a supportive therapy.

Keywords: brg1, ep300, breast cancer, epigenetics

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Authors: Surbhi Surbhi, Andrea Erni, Gunter Meister, Harold Cremer, Christophe Beclin

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MicroRNAs (miRNAs) are short 20-23 nucleotide long non-coding RNAs; there are 2605 miRNA in humans and 1936 miRNA in mouse in total (miRBase). The nervous system expresses the most abundant miRNA and most diverse. MiRNAs play a role in many steps during neurogenesis, like cell proliferation, differentiation, neural patterning, axon pathfinding, etc. Moreover, in vitro studies suggested a role in the regulation of local translation at the synapse, thus controlling neuronal plasticity. However, due to the specific structure of miRNA molecules, an in-vivo confirmation of the general role of miRNAs in the control of neuronal plasticity is still pending. For example, their small size and their high level of sequence homology make difficult the analysis of their cellular and sub-cellular localization in-vivo by in-situ hybridization. Moreover, it was found that only 40% of the expressed miRNA molecules in a cell are included in RNA-Induced Silencing Complexes (RISC) and, therefore, involved in inhibitory interactions while the rest is silent. Definitively, the development of new tools is needed to have a better understanding of the cellular function of miRNAs, in particular their role in neuronal plasticity. Here we describe a new technique called in-vivo AGO-APP designed to investigate miRNA expression and function in-vivo. This technique is based on the expression of a small peptide derived from the human RISC-complex protein TNRC6B, called T6B, which binds all known Argonaute (Ago) proteins with high affinity allowing the efficient immunoprecipitation of AGO-bound miRNAs. We have generated two transgenic mouse lines conditionally expressing T6B either ubiquitously in the cell or targeted at the synapse. A comparison of the repertoire of miRNAs immuno-precipitated from mature neurons of both mouse lines will provide us with a list of miRNAs showing a specific activity at the synapse. The physiological role of these miRNAs will be subsequently addressed through gain and loss of function experiments.

Keywords: RNA-induced silencing complexes, TNRC6B, miRNA, argonaute, synapse, neuronal plasticity, neurogenesis

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Authors: Hamza I. Isa, Gezina C. H. Ferreira, Jan E. Crafford, Christoffel J. Botha

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Moraea pallida Bak. (yellow tulip) poisoning is the most important plant-induced cardiac glycoside toxicosis in South Africa. Cardiac glycoside poisonings collectively account for about 33 and 10 % mortalities due to plants, in large and small stock respectively, in South Africa. The toxic principle is 1α, 2α-epoxyscillirosidine, a bufadienolide. The aim of the study was to investigate the potential to develop a vaccine against epoxyscillirosidine. Epoxyscillirosidine and the related bufadienolides proscillaridin and bufalin, which are commercially available, were conjugated to the carrier proteins [Hen ovalbumin (OVA), bovine serum albumin (BSA) and keyhole limpet haemocyanin (KLH)], rendering them immunogenic. Adult male New Zealand White rabbits were immunized. In Trials 1 and 2, rabbits (n=6) were, each assigned to two groups. Experimental animals (n=3; n=4) were vaccinated with epoxyscillirosidine-OVA conjugate, while the control (n=3; n=2) were vaccinated with OVA, using Freund’s complete and incomplete and Montanide adjuvants, for Trials 1 and 2, respectively. In Trial 3, rabbits (n=15), randomly allocated to 5 equal groups (I, II, III, IV and V), were vaccinated with proscillaridin-BSA, bufalin-BSA, epoxyscillirosidine-KLH, epoxyscillirosidine-BSA conjugates, and BSA respectively, using Montanide as adjuvant. Vaccination was on Days 0, 21 and 42. Additional vaccinations were done on Day 56 and 63 for Trial 1. Vaccination was by intradermal injection of 0.4 ml of the immunogen (4 mg/ml [Trial 1] and 8 mg/ml for Trials 2 and Trial 3, respectively). Blood was collected pre-vaccination and at 3 week intervals following each vaccination. Antibody response was determined using an indirect ELISA. There was poor immune response associated with the dose (0.4 mg per rabbit) and adjuvant used in Trial 1. Antibodies were synthesized against the conjugate administered in Trial 2. For Trail 3, antibodies against the immunogens were successfully raised in rabbits with epoxyscillirosidine-KLH inducing the highest immune response. The antibodies raised against proscillaridin and bufalin cross-reacted with epoxyscillirosidine when used as antigen in the ELISA. The study successfully demonstrated the synthesis of antibodies against the bufadienolide conjugates administered. The cross-reactivity of proscillaridin and bufalin with epoxyscillirosidine could potentially be utilized as alternative to epoxyscillirosidine in future studies to prevent yellow tulp poisoning by vaccination.

Keywords: antibodies , bufadienolides, cross-reactivity, epoxyscillirosidine, Moraea pallida, poisoning

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Authors: Lorenzo Petrucci

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This paper is intended to give insight into the potential use of small-scale off-grid trigeneration systems powered by biogas generated in a dairy farm. The off-grid plant object of analysis comprises a dual-fuel Genset as well as electrical and thermal storage equipment and an adsorption machine. The loads are the different apparatus used in the dairy farm, a household where the workers live and a small electric vehicle whose batteries can also be used as a power source in case of emergency. The insertion in the plant of an adsorption machine is mainly justified by the abundance of thermal energy and the simultaneous high cooling demand associated with the milk-chilling process. In the evaluated operational scenario, our research highlights the importance of prioritizing specific small loads which cannot sustain an interrupted supply of power over time. As a consequence, a photovoltaic and thermal panel is included in the plant and is tasked with providing energy independently of potentially disruptive events such as engine malfunctioning or scarce and unstable supplies of fuels. To efficiently manage the plant an energy dispatch strategy is created in order to control the flow of energy between the power sources and the thermal and electric storages. In this article we elaborate on models of the equipment and from these models, we extract parameters useful to build load-dependent profiles of the prime movers and storage efficiencies. We show that under reasonable assumptions the analysis provides a sensible estimate of the generated energy. The simulations indicate that a Diesel Generator sized to a value 25% higher than the total electrical peak demand operates 65% of the time below the minimum acceptable load threshold. To circumvent such a critical operating mode, dump loads are added through the activation and deactivation of small resistors. In this way, the excess of electric energy generated can be transformed into useful heat. The combination of PVT and electrical storage to support the prioritized load in an emergency scenario is evaluated in two different days of the year having the lowest and highest irradiation values, respectively. The results show that the renewable energy component of the plant can successfully sustain the prioritized loads and only during a day with very low irradiation levels it also needs the support of the EVs’ battery. Finally, we show that the adsorption machine can reduce the ice builder and the air conditioning energy consumption by 40%.

Keywords: hybrid power plants, mathematical modeling, off-grid plants, renewable energy, trigeneration

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Authors: F. Polito, M. Cucinotta, A. Conti, C. Lo Giudice, C. Tomasello, F. Angileri, D. La Torre, M. Aguennouz

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Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors, with an extremely poor prognosis. Telomeres lenght is associated with tumor progression in several type of human cancers and telomere elongation is a common molecular feature of advanced malignancies. Among the telomeric shelterin proteins PTOP is required for telomeric protein complex assembly, telomerase recruitment and activity, and telomere length regulation through a PTOP-telomerase interaction. Previous studies suggest that PTOP upregulation is involved in radioresistance and telomere lengthening in colorectal cancer cells. Moreover, in human osteosarcoma cells PTOP deletion led to telomere shortening, increased apoptosis and radiation sensitivity enhancement. However, to date, little is known about the role of PTOP in progression of glioma cancers. In light of this background aim of the study is to investigate the expression of PTOP in different grades of human glioma and its correlation with the pathological grade of gliomas, grades of malignancy, proliferative activity and apoptosis. Fifteen Low Grade Astrocytomas (LGA), 18 Anaplastic Astrocytomas (AA) and 26 Glioblastoma Multiforme (GBM) samples were analyzed. Three samples of normal brain tissue (NBT) were used as controls. The expression levels of PTOP, h-TERT, BIRC1 and cyclin D1 were determined by real time PCR and/or western blot. Results obtained shows that PTOP expression in glioma tissues is tightly correlated with clinical grade ( p < 0.01 ). No correlation was found between PTOP expression and other clinicopathologic parameters. The expression of PTOP was positively correlated with the expression of hTERT and TERF1. Furthermore PTOP positively correlates with cyclin D1 and negatively correlates with the expression of BIRC1. Our findings indicate that PTOP might play key role in the progression of glioma regulating telomerase activity and likely through regulation of cell cycle and apoptosis. In conclusion results obtained prompted us to speculate that PTOP might represents a potential molecular bio marker and a therapeutic target for the treatment of glioblastoma tumors.

Keywords: glioblastoma, PTOP, telomere, brain tumors

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Authors: Maico R. Severino, Laura G. Caixeta, Nadine M. Costa, Raísa L. T. Napoleão, Éverton F. V. Valle, Diego D. Calixto, Danielle Oliveira

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One of the biggest barriers that companies find nowadays is the coordination of production flow in their Supply Chains (SC). In this study, coordination is understood as a mechanism for incorporating the entire production channel, with everyone involved focused on achieving the same goals. Sometimes, this coordination is attempted by the use of logistics practices or production plan and control methods. No papers were found in the literature that presented the combined use of logistics practices and production plan and control methods. The main objective of this paper is to propose solutions for six case studies combining logistics practices and Ordering Systems (OS). The methodology used in this study was a conceptual model of decision making. This model contains six phases: a) the analysis the types and characteristics of relationships in the SC; b) the choice of the OS; c) the choice of the logistics practices; d) the development of alternative proposals of combined use; e) the analysis of the consistency of the chosen alternative; f) the qualitative and quantitative assessment of the impact on the coordination of the production flow and the verification of applicability of the proposal in the real case. This study was conducted on six Brazilian SC of different sectors: footwear, food and beverages, garment, sugarcane, mineral and metal mechanical. The results from this study showed that there was improvement in the coordination of the production flow through the following proposals: a) for the footwear industry the use of Period Bath Control (PBC), Quick Response (QR) and Enterprise Resource Planning (ERP); b) for the food and beverage sector firstly the use of Electronic Data Interchange (EDI), ERP, Continuous Replenishment (CR) and Drum-Buffer-Rope Order (DBR) (for situations in which the plants of both companies are distant), and secondly EDI, ERP, Milk-Run and Review System Continues (for situations in which the plants of both companies are close); c) for the garment industry the use of Collaborative Planning, Forecasting, and Replenishment (CPFR) and Constant Work-In-Process (CONWIP) System; d) for the sugarcane sector the use of EDI, ERP and CONWIP System; e) for the mineral processes industry the use of Vendor Managed Inventory (VMI), EDI and MaxMin Control System; f) for the metal mechanical sector the use of CONWIP System and Continuous Replenishment (CR). It should be emphasized that the proposals are exclusively recommended for the relationship between client and supplier studied. Therefore, it cannot be generalized to other cases. However, what can be generalized is the methodology used to choose the best practices for each case. Based on the study, it can be concluded that the combined use of OS and logistics practices enable a better coordination of flow production on SC.

Keywords: supply chain management, production flow coordination, logistics practices, ordering systems

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Authors: Jinhee Kang, Kwangman Park, Ruhee Song, Suhee Kim, Do-Hyeon Yu, Kyoungseong Choi, Jinho Park

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Statement of the Problem: Animal blood components maintain homeostasis when animals are healthy, and changes in chemical composition of the blood and body fluids can be observed if animals have a disease. In particular, newborn calves are susceptible to disease and therefore hematologic tests and serum chemistry tests could become an important guideline to the diagnosis and the treatment of diseases. Diarrhea in newborn calves is the most damaging to cattle ranch, whether dairy or cattle fattening, and is a large part of calf atrophy and death. However, since the study on calf electrophoresis was not carried out, a survey analysis was conducted on it. Methodology and Theoretical Orientation: The calves were divided into healthy calves and disease (diarrhea) calves, and calves were classified by 1-14d, 15-28d, and more than 28d, respectively. The fecal state was classified by solid (0-value), semi-solid (1-value), loose (2-value) and watery (3-value). In the solid (0-value) and semi-solid (1-value) feces valuable pathogen was not detected, but loose (2-value) and watery (3-value) feces were detected. Findings: ALB, α-1, α-2, α-SUM, β and γ (Gamma) were examined by electrophoresis analysis of healthy calves and diarrhea calves. Test results showed that there were age differences between healthy calves and diarrheic calves. When we look at the γ-globulin at 1-14 days of age, we can see that the average calf of healthy calves is 16.8% and the average of diarrheal calves is 7.7%, when we look at the figures for the α-2 at 1-14 days, we found that healthy calves average 5.2% and diarrheal calves 8.7% higher than healthy cows. On α-1, 15-28 days, and after 28 days, healthy calves average 10.4% and diarrheal calves average 7.5% diarrhea calves were 12.6% and 12.4% higher than healthy calves. In the α-SUM, the healthy calves were 21.6%, 16.8%, and 14.5%, respectively, after 1-14 days, 15-28 days and 28 days. diarrheal calves were 23.1%, 19.5%, and 19.8%. Conclusion and Significance: In this study, we examined the electrophoresis results of healthy calves and diseased (diarrhea) calves, gamma globulin at 1-14 days of age were lower than those of healthy calves (diarrhea), indicating that the calf was unable to consume colostrum from the mother when it was a new calf. α-1, α-2, α-SUM may be associated with an acute inflammatory response as a result of increased levels of calves with diarrhea (diarrhea). Further research is needed to investigate the effects of acute inflammatory responses on additional calf-forming proteins. Information on the results of the electrophoresis test will be provided where necessary according to the item.

Keywords: alpha, electrophoretogram, serum protein, γ, gamma

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Authors: G. Vici, L. Cesanelli, L. Belli, R. Ceci, V. Polzonetti

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Several factors contribute to success in sport and diet is one of them. Evidence-based sport nutrition guidelines underline the importance of macro- and micro-nutrients’ balance and timing in order to improve athlete’s physical status and performance. Nevertheless, a high content of proteins is commonly found in resistance training athletes’ diet with carbohydrate intake that is not enough or not well planned. The aim of the study was to evaluate the impact of different protein and carbohydrate diet contents on body composition and sport performance on a group of resistance training athletes. Subjects were divided as study group (n=16) and control group (n=14). For a period of 4 months, both groups were subjected to the same resistance training fitness program with study group following a specific diet and control group following an ab libitum diet. Body compositions were evaluated trough anthropometric measurement (weight, height, body circumferences and skinfolds) and Bioimpedence Analysis. Physical strength and training status of individuals were evaluated through the One Repetition Maximum test (RM1). Protein intake in studied group was found to be lower than in control group. There was a statistically significant increase of body weight, free fat mass and body mass cell of studied group respect to the control group. Fat mass remains almost constant. Statistically significant changes were observed in quadriceps and biceps circumferences, with an increase in studied group. The MR1 test showed improvement in study group’s strength but no changes in control group. Usually people consume hyper-proteic diet to achieve muscle mass development. Through this study, it was possible to show that protein intake fixed at 1,7 g/kg/d can meet the individual's needs. In parallel, the increased intake of carbohydrates, focusing on quality and timing of assumption, has enabled the obtainment of desired results with a training protocol supporting a hypertrophic strategy. Therefore, the key point seems related to the planning of a structured program both from a nutritional and training point of view.

Keywords: body composition, diet, exercise, protein

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Authors: Shanker Kalakotla, Krishna Mohan Gottumukkala

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Diabetes is a metabolic disorder characterized by hyperglycemia, carbohydrates, altered lipids and proteins metabolism. In recent research nanotechnology is a blazing field for the researchers; latterly there has been prodigious excitement in the nanomedicine and nano pharmacological area for the study of silver nanoparticles synthesis using natural products. Biological methods have been used to synthesize silver nanoparticles in presence of medicinally active antidiabetic plants, and this intention made us assess the biologically synthesized silver nanoparticles from the seed extract of Psoralea corylfolia using 1 mM silver nitrate solution. The synthesized herbal mediated silver nanoparticles (HMSNP’s) then subjected to various characterization techniques such as XRD, SEM, EDX, TEM, DLS, UV and FT-IR respectively. In current study, the silver nanoparticles tested for in-vitro anti-diabetic activity and possible toxic effects in healthy female albino mice by following OECD guidelines-425. Herbal mediated silver nanoparticles were successfully obtained from bioreduction of silver nitrate using Psoralea corylifolia plant extract. Silver nanoparticles have been appropriately characterized and confirmed using different types of equipment viz., UV-vis spectroscopy, XRD, FTIR, DLS, SEM and EDX analysis. From the behavioral observations of the study, the female albino mice did not show sedation, respiratory arrest, and convulsions. Test compounds did not cause any mortality at the dose level tested (i.e., 2000 mg/kg body weight) doses till the end of 14 days of observation and were considered safe. It may be concluded that LD50 of the HMSNPs was 2000mg/kg body weight. Since LD50 of the HMSNPs was 2000mg/kg body weight, so the preferred dose range for HMSNPs falls between the levels of 200 and 400 mg/kg. Further In-vivo pharmacological models and biochemical investigations will clearly elucidate the mechanism of action and will be helpful in projecting the currently synthesized silver nanoparticles as a therapeutic target in treating chronic ailments.

Keywords: herbal mediated silver nanoparticles, HMSNPs, toxicity of silver nanoparticles, PTP1B in-vitro anti-diabetic assay female albino mice, 425 OECD guidelines

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Authors: Amel Fayed, Shaden AlSubaih, Nouf Al-Qahtani, Asmaa Gosty, Asma Aljuhaimi

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Introduction: Sleep difficulties are vastly predominant among adults and affect different aspects of their life. Many literatures found out that females are more liable to suffer from sleeping problems. College students are typical example of people dealing with daily pressure and stress to fulfill the daily tasks and responsibilities. In addition to their ultimate goal of achieving excellent academic records which require their full concentration and effort. Consequently, many of them start complaining of sleep deprivations which can undesirably affect their academic achievements. This study was aiming to investigate how prevalent is sleeping disorders among different colleges in the university and its relation their academic achievements. Methods: A cross-sectional study of female university students at Princess Norah Bint Abdulrahman University using self-administered questionnaire was conducted. Insomnia Severity Index (ISI) was used to assess different grades of insomnia. Students were requested to answer the questions evaluating their sleeping habits over the last two weeks. Participants reported their latest Grade Point Average (GPA). According to ISI, insomnia severity is reported as ‘No clinically significant’, ‘Subthreshold ‘,’ Clinical moderate insomnia’ and ‘Clinical severe’. Results: In the current study, 228 students participated; 172(75.4%) from medical colleges and 56 (24.6%) from non-medical colleges. About 80% of them claimed to have never taken any medications to help them sleep while only three students confirmed their regular use of sleep-inducing medications. About 16% of the students drink milk or other hot drinks to help them fall asleep. None of the students was suspected of having obstructive sleep apnea or apparent psychiatric disorder. According to ISI, 182 (79.8%) students suffered from subthreshold insomnia, 37 (16.2%) had clinical insomnia (moderate severity) and 9 (3.9%) of students had sleeping problems of non-clinically significance level. However, none of students was found to have severe clinical insomnia. Clinical moderate insomnia was reported in 15.1% of medical students and 19.6% of non-medical students. Moreover, about 82% of medical students suffered from subthreshold insomnia compared to 73.2% of non-medical students. This difference was not statistically significant (P=0.24). About 63% of medical students and 48% of non-medical students believed that high percentage of their colleagues are suffering from insomnias (p-value 0.08) The association between GPA and insomnia revealed that; 19.5% of low GPA group compared to 9.3% of high GPA group had clinical moderate insomnia. This association was not statistically significant (p=0.15). The correlation between the GPA and the ISI score was negative but not conclusive (r=-0.08, p-value = 0.29). More than 92% of all students agreed that sleeping problems affect their academic achievement to varying degrees. Conclusion: our results suggest that insomnia is commonly prevalent among female university students and might affect the students’ achievement. This study provides preliminary data about the quality of sleep among medical and non-medical university students which may be used to promote the healthy sleeping habits among female students.

Keywords: academic achievement, females, insomnia, university student

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Authors: Sneha Singh, Abhimanyu Dev, Vinod Nigam

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The major issue which decides the impending use of gold nanoparticles (AuNPs) in nanobiotechnological applications is their particle size and stability. Often the AuNPs obtained biomimetically are considered useless owing to their instability in the aqueous medium and thereby limiting the widespread acceptance of this facile green synthesis procedure. So, the use of nontoxic surfactants is warranted to stabilize the biogenic nanoparticles (NPs). But does the surfactant only play a role in stabilizing by being adsorbed to the NPs surface or can it have any other significant contribution in synthesis process and controlling their size as well as shape? Keeping this idea in mind, AuNPs were synthesized by using surfactant treated (lechate) and untreated (cell lysate supernatant) Bacillus licheniformis cell extract. The cell extracts mediated reduction of chloroauric acid (HAuCl 4) in the presence of non-ionic surfactant, Tween 20 (TW20), and its effect on the AuNPs stability was studied. Interestingly, the surfactant used in the study served as potential alternative to harvest cellular enzymes involved in bioreduction process in a hassle free condition. The surfactants ability to solubilize/leach membrane proteins and simultaneously stabilizing the AuNPs could have advantage from process point of view as it will reduce the time and economics involve in the nanofabrication of biogenic NPs. The synthesis was substantiated with UV-Vis spectroscopy, Dynamic light scattering study, FTIR spectroscopy, and Transmission electron microscopy. The Zeta potential of AuNPs solutions was measured routinely to corroborate the stability observations recorded visually. Highly stable, ultra-small AuNPs of 2.6 nm size were obtained from the study. Further, the biological efficacy of the obtained AuNPs as potential antibacterial agent was evaluated against Bacilllus subtilis, Pseudomonas aeruginosa, and Escherichia coli by observing the zone of inhibition. This potential of AuNPs of size < 3 nm as antibacterial agent could pave way for development of new antimicrobials and overcoming the problems of antibiotics resistance

Keywords: antibacterial, bioreduction, nanoparticles, surfactant

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Authors: Muhammad Ali Siddiquee, Md. Alauddin, Md. Omar Faruque, Zakir Hossain Howlader, Mohammad Asaduzzaman

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Rice bran (RB) and rice bran oil (RBO) are explored as prominent food components worldwide. In this study, fermented rice bran (FRB) was produced by employing edible gram-positive bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus, and Bifidobacterium bifidum) at 125 x 10⁵ spore g⁻¹ of rice bran, and investigated to evaluate nutritional quality. The crude rice bran oil (CRBO) was extracted from RB, and its quality was also investigated compared to market-available rice bran oil (MRBO) in Bangladesh. We found that fermentation of rice bran with lactic acid bacteria increased total proteins (29.52%), fat (5.38%), ash (48.47%), crude fiber (38.96%), and moisture (61.04%) and reduced the carbohydrate content (36.61%). We also found that essential amino acids (methionine, tryptophan, threonine, valine, leucine, lysine, histidine, and phenylalanine) and non-essential amino acids (alanine, aspartate, glycine, glutamine, proline, serine, and tyrosine) were increased in FRB except methionine and proline. Moreover, total phenolic content, tannin content, flavonoid content, and antioxidant activity were increased in FRB. The RBO analysis showed that γ-oryzanol content (10.00mg/g) was found in CRBO compared to MRBO (ranging from 7.40 to 12.70 mg/g) and Vitamin-E content 0.20% was found higher in CRBO compared to MRBO (ranging 0.097 to 0.12%). The total saturated (25.16%) and total unsaturated fatty acids (74.44%) were found in CRBO, whereas MRBO contained total saturated (22.08 to 24.13%) and total unsaturated fatty acids (71.91 to 83.29%), respectively. The physiochemical parameters were found satisfactory in all samples except acid value and peroxide value higher in CRBO. Finally, animal experiments showed that FRB and CRBO reduce the body weight, glucose, and lipid profile in high-fat diet-induced animal models. Thus, FRB and RBO could be value-added food supplements for human health.

Keywords: fermented rice bran, crude rice bran oil, amino acids, proximate composition, gamma-oryzanol, fatty acids, heavy metals, physiochemical parameters

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Authors: Zeiler M., Stadler D., Schmaderer C.

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Over the years of dialysis treatment, most patients experience significant weight loss. The primary emphasis in earlier research was the underlying mechanism of protein energy wasting and the subsequent malnutrition inflammation syndrome. In the interest to provide an effective and rapid solution for the patients, the aim of this study is identifying individual influences of their assumed reduced dietary intake, such as nausea, appetite loss and taste changes, and to determine whether the patients adhere to their nutrition guidelines. A prospective, controlled study with 38 end-stage renal disease patients was performed using a questionnaire to reflect their diet within the last 12 months. Thereby, the daily intake for the most important macro-and micronutrients was calculated to be compared with the individual KDQOI-guideline value, as well as controls matched in age and gender. The majority of the study population did not report symptoms commonly associated with dialysis, such as nausea or inappetence, and denied any change in dietary behavior since receiving renal replacement therapy. The patients’ daily intake of energy (3080kcal ± 1266) and protein (89,9g [53,4-142,0]) did not differ significantly from the controls (energy intake: 3233kcal ± 1046, p=0,597; protein intake: 103,7g [90,1-125,5], p=0,120). The average difference to the individual calculated KDQOI-guideline was +176,0kcal ± 1156 (p=0,357) for energy intake and -1,75g ± 45,9 (p=0,491) for protein intake. However, there was an observed imbalance in the distribution of macronutrients, with a preference for fats over proteins. The patients’ daily intake of sodium (5,4g [ 2,95-10,1]) was higher than in the controls (4,1g [2,04-5,99], p= 0,058) whereas both values for potassium (3,7g ± 1,84) and phosphorous (1,79g ± 0,91) went significantly below the controls’ values (potassium intake: 4,89g ± 1,74, p=0,014; phosphorous intake: 2,04g ± 0,64, p=0,038). Thus, the values exceeded the calculated KDQOI-recommendation by + 3,3g [0,63-7,90] (p<0,001) for sodium, +1,49g ± 1,84 (p<0,001) for potassium and +0,89g ± 0,91 (p<0,001) for phosphorous. Contrary to the assumption, the patients did not under-eat. Nevertheless, their diets did not align with the recommended values. These findings highlight the need for intervention and education among patients and that regular dietary monitoring could prevent unhealthy nutrition habits. The elaboration of individual references instead of standardized guidelines could increase the compliance to the advised diet so that interdisciplinary comorbidities do not develop or worsen.

Keywords: compliance, dialysis, end-stage renal disease, KDQOI, malnutrition, nutrition guidelines, questionnaire, salt intake

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Authors: Fouad Chebib, Marie Hogan, Ziad El-Zoghby, Maria Irazabal, Sarah Senum, Christina Heyer, Charles Madsen, Emilie Cornec-Le Gall, Atta Behfar, Barbara Ehrlich, Peter Harris, Vicente Torres

Abstract:

Background: Mutations in PKD1 and PKD2, the genes encoding the proteins polycystin-1 (PC1) and polycystin-2 (PC2) cause autosomal dominant polycystic kidney disease (ADPKD). ADPKD is a systemic disease associated with several extrarenal manifestations. Animal models have suggested an important role for the polycystins in cardiovascular function. The aim of the current study is to evaluate the association of various cardiomyopathies in a large cohort of patients with ADPKD. Methods: Clinical data was retrieved from medical records for all patients with ADPKD and cardiomyopathies (n=159). Genetic analysis was performed on available DNA by direct sequencing. Results: Among the 58 patients included in this case series, 39 patients had idiopathic dilated cardiomyopathy (IDCM), 17 had hypertrophic obstructive cardiomyopathy (HOCM), and 2 had left ventricular noncompaction (LVNC). The mean age at cardiomyopathy diagnosis was 53.3, 59.9 and 53.5 years in IDCM, HOCM and LVNC patients respectively. The median left ventricular ejection fraction at initial diagnosis of IDCM was 25%. Average basal septal thickness was 19.9 mm in patients with HOCM. Genetic data was available in 19, 8 and 2 cases of IDCM, HOCM, and LVNC respectively. PKD1 mutations were detected in 47.4%, 62.5% and 100% of IDCM, HOCM and LVNC cases. PKD2 mutations were detected only in IDCM cases and were overrepresented (36.8%) relative to the expected frequency in ADPKD (~15%). The prevalence of IDCM, HOCM, and LVNC in our ADPKD clinical cohort was 1:17, 1:39 and 1:333 respectively. When compared to the general population, IDCM and HOCM was approximately 10-fold more prevalent in patients with ADPKD. Conclusions: In summary, we suggest that PKD1 or PKD2 mutations may predispose to idiopathic dilated or hypertrophic cardiomyopathy. There is a trend for patients with PKD2 mutations to develop the former and for patients with PKD1 mutations to develop the latter. Predisposition to various cardiomyopathies may be another extrarenal manifestation of ADPKD.

Keywords: autosomal dominant polycystic kidney (ADPKD), polycystic kidney disease, cardiovascular, cardiomyopathy, idiopathic dilated cardiomyopathy, hypertrophic cardiomyopathy, left ventricular noncompaction

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