Search results for: protein isolates

Authors: Tahra ElObeid, Omnya Ahmed, Reem Al-Sharshani, Doaa Dalloul, Jannat Alnattei

Abstract:

Background: Camelus dormedarius camels are also called ‘the Arabian camels’ and are present in the desert area of North Africa and the Middle East. Recently, camel’s milk has a great attention globally because of their proteins and peptides that have been reported to be beneficial for the health and in the management of many diseases. Objectives: This study was designed to investigate the antioxidant, antimicrobial activity and to evaluate the total phenolic content of camel’s milk proteins in Qatar. Methods: Fresh two camel’s milk samples from Omani breed and called Muhajer (camel’s milk A and B) were collected on the 1st of the December. Both samples were from the same location Al- Shahaniyah, Doha, Qatar, but from different local private farms and feeding system. Camel’s milk A and B were defatted by centrifugation and their proteins were extracted by acid and thermal precipitation. The antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Total phenolic compound (TPC) was evaluated by Folin-Ciocalteu reagent (FCR). On the other hand, the antimicrobial activity against eight different type of pathogenic bacteria was evaluated by disc diffusion method and the zone of inhibition was measured. Results: The of the total phenolic content of whole milk in both camel’s milk A and B were significantly the highest among the protein extracts. The % of the DPPH radical inhibition of casein protein in both camel’s milk A and B were significantly the highest among the protein extracts. In this study, there were marked changes in the antibacterial activity in the different camel milk protein extracts. All extracts showed bacterial overgrowth. Conclusion: The antioxidant activity of the camel milk protein extracts correlated to their unique phenolic compounds and bioactive protein peptides. The antimicrobial activity was not detected perhaps due to the technique, the quality, or the extraction method. Overall, camel's milk exhibits a high antioxidant activity, which is responsible for many health benefits besides the nutritional values.

Keywords: camels milk, antioxidant content, antimicrobial activity, proteins, Qatar

Procedia PDF Downloads 187

Authors: Sima E. Rugarabamu, Mecky I. Matee, Elison N. M. Simon

Abstract:

Background: In Tanzania bacteriological studies of etiological agents of oro-facial infections are very limited, and very few have investigated anaerobes. The aim of this study was to determine the spectrum of bacterial agents involved in oral and maxillofacial infections in patients attending Muhimbili National Hospital, Dar-es-salaam, Tanzania. Method: This was a hospital based descriptive cross-sectional study that was conducted in the Department of Oral and Maxillofacial Surgery of the Muhimbili National Hospital in Dar es Salaam, Tanzania from 1st January 2014 to 31st August 2014. Seventy (70) patients with various forms of oral and maxillofacial infections who were recruited for the study. The study participants were interviewed using a prepared questionnaire after getting their consent. Pus aspirate was cultured on Blood agar, Chocolate Agar, MacConkey agar and incubated aerobically at 37°C. Imported blood agar was used for anaerobic culture whereby they were incubated at 37°Cin anaerobic jars in an atmosphere of generated using commercial gas-generating kits in accordance with manufacturer’s instructions. Plates were incubated at 37°C for 24 hours (For aerobic culture and 48 hours for anaerobic cultures). Gram negative rods were identified using API 20E while all other isolates were identified by conventional biochemical tests. Antibiotic sensitivity testing for isolated aerobic and anaerobic bacteria was detected by the disk diffusion, agar dilution and E-test using routine and commercially available antibiotics used to treat oral facial infections. Results: This study comprised of 41 (58.5%) males and 29 (41.5%) females with a mean age of 32 years SD +/-15.1 and a range of 19 to 70 years. A total of 161 bacteria strains were isolated from specimens obtained from 70 patients which were an average of 2.3 isolates per patient. Of these 103 were aerobic organism and 58 were strict anaerobes. A complex mix of strict anaerobes and facultative anaerobes accounted for 87% of all infections.The most frequent aerobes isolated was streptococcus spp 70 (70%) followed by Staphylococcus spp 18 (18%). Other organisms such as Klebsiella spp 4 (4%), Proteus spp 5 (5%) and Pseudomonas spp 2 (2%) were also seen. The anaerobic group was dominated by Prevotella spp 25 (43%) followed by Peptostreptococcus spp 18 (31%); other isolates were Pseudomonas spp 2 (1%), black pigmented Pophyromonas spp 4 (5%), Fusobacterium spp 3 (3%) and Bacteroides spp 5 (8%). Majority of these organisms were sensitive to Amoxicillin (98%), Gentamycin (89%), and Ciprofloxacin (100%). A 40% resistance to metronidazole was observed in Bacteroides spp otherwise this drug and others displayed good activity against anaerobes. Conclusions: Oral and maxillofacial facial infections at Muhimbili National Hospital are mostly caused by streptococcus spp and Prevotella spp. Strict anaerobes accounted for 36% of all isolates. The profile of isolates should assist in selecting empiric therapy for infections of the oral and maxillofacial region. Inclusion of antimicrobial agents against anaerobic bacteria is highly recommended.

Keywords: bacteria, oral and maxillofacial infections, antibiotic susceptibility, Tanzania

Procedia PDF Downloads 308

Authors: Seyed Reza Aghili, Tahereh Shokohi, Shirin Sadat Hashemi Fesharaki, Mohammad Ali Boroumand, Bahar Salmanian

Abstract:

Introduction: Intravenous catheter-associated fungal infection as nosocomial infection continue to be a deep problem among hospitalized patients, decreasing quality of life and adding healthcare costs. The capacity of catheters in the spread of candidemia in heart failure patients is obvious. The aim of this study was to evaluate the prevalence and genetic identification of Candida species in heart disorder patients. Material and Methods: This study was conducted in Tehran Hospital of Cardiology Center (Tehran, Iran, 2014) during 1.5 years on the patients hospitalized for at least 7 days and who had central or peripheral vein catheter. Culture of catheters, blood and skin of the location of catheter insertion were applied for detecting Candida colonies in 223 patients. Identification of Candida species was made on the basis of a combination of various phenotypic methods and confirmed by sequencing the ITS1-5.8S-ITS2 region amplified from the genomic DNA using PCR and the NCBI BLAST. Results: Of the 223 patients samples tested, we identified totally 15 Candida isolates obtained from 9 (4.04%) catheter cultures, 3 (1.35%) blood cultures and 2 (0.90%) skin cultures of the catheter insertion areas. On the base of ITS region sequencing, out of nine Candida isolates from catheter, 5(55.6%) C. albicans, 2(22.2%) C. glabrata, 1(11.1%) C. membranifiaciens and 1 (11.1%) C. tropicalis were identified. Among three Candida isolates from blood culture, C. tropicalis, C. carpophila and C. membranifiaciens were identified. Non-candida yeast isolated from one blood culture was Cryptococcus albidus. One case of C. glabrata and one case of Candida albicans were isolated from skin culture of the catheter insertion areas in patients with positive catheter culture. In these patients, ITS region of rDNA sequence showed a similarity between Candida isolated from the skin and catheter. However, the blood samples of these patients were negative for fungal growth. We report two cases of catheter-related candidemia caused by C. membranifiaciens and C. tropicalis on the base of genetic similarity of species isolated from blood and catheter which were treated successfully with intravenous fluconazole and catheter removal. In phenotypic identification methods, we could only identify C. albicans and C. tropicalis and other yeast isolates were diagnosed as Candida sp. Discussion: Although more than 200 species of Candida have been identified, only a few cause diseases in humans. There is some evidence that non-albicans infections are increasing. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition are considered to be associated with candidemia in hospitalized heart failure patients. Identifying the route of infection in candidemia is difficult. Non-albicans candida as the cause of candidemia is increasing dramatically. By using conventional method, many non-albicans isolates remain unidentified. So, using more sensitive and specific molecular genetic sequencing to clarify the aspects of epidemiology of the unknown candida species infections is essential. The positive blood and catheter cultures for candida isolates and high percentage of similarity of their ITS region of rDNA sequence in these two patients confirmed the diagnosis of intravenous catheter-associated candidemia.

Keywords: catheter-associated infections, heart failure patient, molecular genetic sequencing, ITS region of rDNA, Candidemia

Procedia PDF Downloads 305

Authors: Omaima A. Sharaf, Tarek A. Moussa, Said M. Badr El-Din, H. Moawad

Abstract:

Background: Polycyclic aromatic hydrocarbons (PAHs) pose great environmental and human health concerns for their widespread occurrence, persistence, and carcinogenic properties. PAHs releases due to anthropogenic activities to the wider environment have led to higher concentrations of these contaminants than would be expected from natural processes alone. This may result in a wide range of environmental problems that can accumulate in agricultural ecosystems, which threatened to become a negative impact on sustainable agricultural development. Thus, this study aimed to evaluate the physico-chemical, and microbial properties of the compost leachate (CL) to assess its role as nutrient and microbial source (biostimulation/bioaugmentation) for developing a cost-effective bioremediation technology for PAHs contaminated sites. Material and Methods: PAHs-degrading bacteria were isolated from CL that was collected from a composting site located in central Scotland, UK. Isolation was carried out by enrichment using phenanthrene (PHR), pyrene (PYR) and benzo(a)pyrene (BaP) as the sole source of carbon and energy. The isolates were characterized using a variety of phenotypic and molecular properties. Six different isolates were identified based on the difference in morphological and biochemical tests. The efficiency of these isolates in PAHs utilization was assessed. Further analysis was performed to define taxonomical status and phylogenic relation between the most potent PAHs-utilizing bacterial strains and other standard strains, using molecular approach by partial 16S rDNA gene sequence analysis. Results indicated that the 16S rDNA sequence analysis confirmed the results of biochemical identification, as both of biochemical and molecular identification of the isolates assigned them to Bacillus licheniformis, Pseudomonas aeruginosa, Alcaligenes faecalis, Serratia marcescens, Enterobacter cloacae and Providenicia which were identified as the prominent PAHs-utilizers isolated from CL. Conclusion: This study indicates that the CL samples contain a diverse population of PAHs-degrading bacteria and the use of CL may have a potential for bioremediation of PAHs contaminated sites.

Keywords: polycyclic aromatic hydrocarbons, physico-chemical analyses, compost leachate, microbial and biochemical analyses, phylogenic relations, 16S rDNA sequence analysis

Procedia PDF Downloads 247

Authors: Ravinder Singh, Ankita Gurao, Saroj Bandhan, Sudhir Kumar Kashyap

Abstract:

The Bos taurus Beta defensin 3 is a defensin peptide secreted by neutrophils and epithelial that exhibits anti-microbial activity. It is one of the crucial components forming an innate defense against intra mammary infections in livestock. The beta defensin 3 by virtue of its anti-microbial activity inhibits major mastitis pathogens including Staphylococcus aureus and Pseudomonas aeruginosa etc, which are also responsible for biofilm formation leading to antibiotic resistance phenomenon. Therefore, the defensin may prove as a non-conventional option to treat mastitis. In this study, computational analysis has been performed including sequence comparison among species and homology modeling of Bos taurus beta defensin 3 protein. The assessments of protein structure were done using the protein structure and model assessment tools integrated in Swiss Model server, which employs various local and global quality evaluation parameters. Further, molecular docking was also carried out between the defensin peptide and the components of biofilm to gain insight into various interactions and structural differences crucial for functionality of this protein.

Keywords: beta defensin 3, bos taurus, docking, homology modeling

Procedia PDF Downloads 267

Authors: Ahreum Choi, Otgontuya Tsogbadrakh, Kwang-Hwan Jung

Abstract:

Microbial rhodopsins are well-known seven-transmembrane proteins that have been extensively studied. These retinal-binding proteins have divided into two types. The type I is microbial rhodopsin, and type II (visual pigment) is expressed mostly in mammalian eyes. For type I rhodopsin, there are two main functions that are ion pumping activity and sensory transduction. Anabaena sensory rhodopsin (ASR) is one of the microbial rhodopsin with main function as photo-sensory transduction. Although ASR is expressed fairly well in Escherichia coli, the expression level is relatively less compare to Proteorhodopsin. In this study, full length of ASR was used to test for the expression influence by codon usage in E. coli. Eight amino acids of codon at N-terminal part of ASR were changed randomly with designed primers, which allow 8,192 nucleotide different cases. The codon changes were screened for the preferable codons of each residue, which have given higher expression yield. Among those 57 selected mutations, there are 24 color-enhanced E. coli colonies that contain ASR proteins, and it showed better expression level than the wild type ASR codon usage. This strongly suggests that high codon usage of only partial N-terminal of protein can increase the expression level of whole protein.

Keywords: 7-transmembrane, all-trans retinal, rhodopsin, codon-usage, protein expression

Procedia PDF Downloads 161

Authors: D. S. V. Barbieri, R. R. Gomes, G. D. Santos, P. F. Herkert, M. Moreira, E. S. Trindade, V. A. Vicente

Abstract:

The importance of yeast infections has increased in recent decades. The monitoring of Candida yeasts has been relevant in the study of groups and populations. This research evaluated 31 Candida spp. isolates from oral microbiota of 12 Brazilian schoolchildren coinfected with Streptococcus mutans. The isolates were evaluated for their ability to form biofilm in vitro and molecularly characterized based on the sequencing of intergenic spacer regions ITS1-5,8S-ITS2 and variable domains of the large subunit (D1/D2) regions of the rDNA, as well as ABC system genotyping. The sequencing confirmed 26 lineages of Candida albicans, three Candida tropicalis, one Candida guillhermondii and one Candida glabrata. Genetic variability and differences on in biofilm formation were observed among Candida yeasts lineages. At least one Candida strain from each caries activity child was C.albicans genotype A or Candida non-albicans. C. tropicalis was associated with highest cavities rates. These results indicate that the presence of C. albicans genotype A or multi-colonization by non albicans species seem to be associates to the potentialization of caries risk.

Keywords: biofilm, Candida albicans, oral microbiota, caries

Procedia PDF Downloads 489

Authors: Latifat Afolake Ogunfolabo, Hakeem Babafemi Ogunfolabo

Abstract:

The gastrointestinal tract is densely populated with micro-organism which closely and intensively interacts with the host and ingested feed. Food borne infections are some of the major international challenges that lead to high mortality and also, antimicrobial resistance, which has been classified as a serious threat by World Health Organization. Samples of slaughtered cattle and goats intestines were collected and standard culture methods were used for bacteria isolation and identification. Minimum inhibitory concentration of commonly used antibiotic using modification of the disk diffusion method was carried out on isolates. The samples cultured were all positive to Pseudomonas aeruginosa (95% and 90%), Escherichia coli (85%), Salmonella typhi (70% and 60%), Staphylococcus aureus (75%and 100%), Micrococcus luteus (55% and35%), Bacillus macerans (60% and 5%), Bacillus cereus (25% and 20%), Clostridium perfringens (20% and 5%), Micrococcus varians (20% and 5%), Bacillus subtilis (25% and 5%), Streptococcus faecalis (40% and 25%) and Streptococcus faecium (15% and 10%) in goat and cattle respectively. Also, Proteus mirabilis (40%), Micrococcus luteus (35%), Proteus vulgaris (30%), Klebsiella aerogenes(15%) were isolated from cattle. The total coliform (13.55 x10⁵cfu/gm ± 1.77) and (20.30 x10⁵cfu/gm ± 1.27) counts were significantly higher than the total bacteria count (8.3 x10⁵cfu/gm ± 1.41) and (16.60 x10⁵cfu/gm ±0.49) for goat and cattle respectively. Selected Bacteria count of isolates showed that Staphylococcus aureus had the highest significant value (6.9 x10⁵cfu/gm ± 0.57) and (16.80 x10⁵cfu/gm ± 0.57) Escherichia coli (4.60 x10⁵cfu/gm ± 0.42) and (7.05 x10⁵cfu/gm ± 0.64) while the lowest significant value was obtained in Salmonella/Shigella (1.7 x10⁵cfu/gm ± 0.00) and (1.5 x10⁵cfu/gm ± 0.00) for goat and cattle respectively. Susceptibility of bacteria isolated from slaughtered goat and cattle intestine to commonly used antibiotics showed that the highest statistical significant value for zone of inhibition for goat was obtained for Ciprofloxacin (30.00 ± 2.25, 23.75 ± 2.49, 17.17 ± 1.40) followed by Augmentin (28.33 ± 1.22, 21. 83 ± 2.44, 16.67 ± 1.49), Erythromycin (27.75 ±1.48, 20.25 ± 1.29, 16.67 ± 1.26) while the lowest values were obtained for Ofloxacin (27.17 ± 1.89, 21.42 ± 2.19, 16.83 ± 1.26) respectively and values obtained for cattle are Ciprofloxacin (30.64 ± 1.6, 25.79 ± 1.76, 8.07 ± 11.49) followed by Augmentin (28.29 ± 1.33, 22.64 ± 1.82, 17.43 ± 1.55) Ofloxacin (26.57 ± 2.02, 20.79 ± 2.75, 16.21 ± 1.19) while the lowest values were obtained for Erythromycin (26.64 ± 1.49, 20.29 ± 1.49, 16.29 ± 1.33) at different dilution factor (10⁻¹, 10⁻², 10⁻³) respectively. The isolates from goat and cattle were all susceptible to Augmentin at the three different dilution factors. Some goat isolates are intermediate to Ciprofloxacin and Erythromycin at 10⁻² and 10⁻³, while resistance to Ciprofloxacin at 10⁻³ dilution factor. Ciprofloxacin and Ofloxacin at the dilution factors of 10⁻³ and 10⁻¹ for some cattle isolate and resistance were observed for Ofloxacin and Erythromycin at dilution of 10⁻³. These results indicate the susceptibilities and the antimicrobial resistance to commonly used antibiotic.

Keywords: antibiotic susceptibility, bacteria, cattle, goat, identification

Procedia PDF Downloads 98

Authors: Vijay Kumar Kutala, Addepalli Pavani, M. Amresh Rao, Naushad Sm

Abstract:

In this study, we evaluated the impact of CYP2C9*2 and CYP2C9*3 variants on binding and hydroxylation of warfarin. In silico data revealed that warfarin forms two hydrogen bonds with protein backbone i.e. I205 and S209, one hydrogen bond with protein side chain i.e. T301 and stacking interaction with F100 in CYP2C9*1. In CYP2C9*2 and CYP2C9*3 variants, two hydrogen bonds with protein backbone are disrupted. In double variant, all the hydrogen bonds are disrupted. The distances between C7 of S-warfarin and Fe-O in CYP2C9*1, CYP2C9*2, CYP2C9*3 and CYP2C9*2/*3 were 5.81A°, 7.02A°, 7.43° and 10.07°, respectively. The glide scores (Kcal/mol) were -7.698, -7.380, -6.821 and -6.986, respectively. Increase in warfarin/7-hydroxy warfarin ratio was observed with increase in variant alleles. To conclude, CYP2C9*2 and CYP2C9*3 variants result in disruption of hydrogen bonding interactions with warfarin and longer distance between C7 and Fe-O thus impairing warfarin 7-hydroxylation due to lower binding affinity of warfarin.

Keywords: warfarin, CYP2C9 polymorphism, personalized medicine, in Silico

Procedia PDF Downloads 296

Authors: Eleni Poulou-Sidiropoulou, Charalampos N. Bompas, Martina Samiotaki, Alexios Vlamis-Gardikas

Abstract:

The glutaredoxin (Grx) and thioredoxin (Trx) systems keep the intracellular environment reduced in almost all organisms. In Escherichia coli (E. coli), the Grx system relies on NADPH+ to reduce GSH reductase (GR), the latter reducing oxidized diglutathione to glutathione (GSH) which in turn reduces cytosolic Grxs, the electron donors for different intracellular substrates. In the Trx system, GR and GSH are replaced by Trx reductase (TrxR). Three of the Grxs of E. coli (Grx1, 2, 3) are reduced by GSH, while Grx4 is likely reduced by TrxR. Trx1 and Grx1 from E. coli may reduce ribonucleotide reductase Ia to ensure a constant supply of deoxyribonucleotides for the synthesis of DNA. The role of the other three Grxs is relatively unknown, especially for Grx2 that may amount up to 1 % of total cellular protein in the stationary phase of growth. The protein is known as a potent antioxidant, but no specific functions have been attributed to it. Herein, affinity chromatography of cellular extracts on immobilized Grx2, followed by MS analysis of the resulting eluates, was employed to identify protein ligands that could provide insights into the biological role of Grx2. Ionic, strong non-covalent, and covalent (disulfide) interactions with relevant proteins were detected. As a means of verification, the identified ligands were subjected to in silico docking with monothiol Grx2. In other experiments, protein extracts from E. coli cells lacking the gene for Grx2 (grxB) were compared to those of wild type. Taken together, the two approaches suggest that Grx2 is involved in protein synthesis, nucleotide metabolism, DNA damage repair, stress responses, and various metabolic processes. Grx2 appears as a versatile protein that may participate in a wide range of biological pathways beyond its known general antioxidant function.

Keywords: Escherichia coli, glutaredoxin 2, interactome, thiol-disulfide oxidoreductase

Procedia PDF Downloads 26

Authors: Shiva Emami, Jenny Persson, Bengt Johansson Lindbom

Abstract:

Group A streptococcus (GAS) is a prevalent human pathogen, causing a wide range of infections and diseases. One of the most well-known virulence factors in GAS is M protein, a surface protein that facilitates bacterial invasion. In this study, we used a recombinant GAS strain (GAS-2W) expressing M protein containing a hyper immunogenic peptide (2W). Mice were immunized three times with heat-killed-GAS subcutaneously at three weeks intervals. Three weeks post last immunization, mice were challenged intraperitoneally with a lethal dose of live GAS. In order to investigate the impact of IFN-ƴ and antibodies in protection against GAS infection, we used a mouse model knock-out for IFN-ƴ (IFN-ƴ KO). We observed immunization with GAS-2W strain can increase protection against GAS infection in mice compared with the original GAS strain. Higher levels of antibodies against M1 protein were measured in GAS-2W-immunized mice. There was also a significant increase in IgG2c response in mice immunized with GAS2W. By using IFN-ƴ KO mice, we showed that not a high level of total IgG, but IgG2c was correlated with protection through the i.p challenge. It also emphasizes the importance of IFN-ƴ cytokine to combat GAS by isotype switching to IgG2c (which is opsonic for phagocytosis). Our data indicate the crucial role of IFN-ƴ in the protective immune response that, together with IgG2c, can induce protection against GAS.

Keywords: Group A streptococcus, IgG2c, IFN-γ, protection

Procedia PDF Downloads 66

Authors: Ravinder Singh, C Rajesh, Saroj Badhan, Shailendra Mishra, Ranjit Singh Kataria

Abstract:

Heat shock protein 60 and 70 are crucial chaperones that guide appropriate folding of denatured proteins under heat stress conditions. HSP60 and HSP70 provide assistance in correct folding of a multitude of denatured proteins. The heat shock factors are the family of some transcription factors which controls the regulation of gene expression of proteins involved in folding of damaged or improper folded proteins during stress conditions. Under normal condition heat shock proteins bind with HSF-1 and act as its repressor as well as aids in maintaining the HSF-1’s nonactive and monomeric confirmation. The experimental protein structure for all these proteins in Bubalus bubalis is not known till date. Therefore computational approach was explored to identify three-dimensional structure analysis of all these proteins. In this study, an extensive in silico analysis has been performed including sequence comparison among species to comparative modeling of Bubalus bubalis HSP60, HSP70 and HSF-1 protein. The stereochemical properties of proteins were assessed by utilizing several scrutiny bioinformatics tools to ensure model accuracy. Further docking approach was used to study interactions between Heat shock proteins and HSF-1.

Keywords: Bubalus bubalis, comparative modelling, docking, heat shock protein

Procedia PDF Downloads 295

Authors: Farnaz Firuzian, Farhad Choobdar, Ali Mazouri

Abstract:

As Very Low Birth Weight (VLBW) neonates cannot tolerate enteral feeding, parenteral nutrition (PN) must be administered shortly after birth. To find an optimal combination of nutrition, in this study, we compare administering high glucose versus high protein intake as a component of total parenteral nutrition (TPN) to test their effect on birth weight (BW) regain in VLBW. This study employs a multi-arm randomized trial: 145 newborns with BW < 1500 g were randomized to control (C) or experimental groups: high glucose (G) or high protein (P). All samples in each group received the same TPN regimens except glucose and protein intake: Glocuse was provided by dextrose water (DW) serum: 7-15 g/kg/d (10% DW) in groups C and P versus 8.75-18.75 g/kg/d (12.5% DW) in group G. Protein provided by amino acids 3 g/kg/d for groups C and G versus 4 g/kg/d for group P. Outcomes (weight, height, and head circumference) was monitored on a daily basis until the BW was regained. Data has been gathered recently and is being processed. We hypothesize that neonates with higher amino acid intake will result in sooner BW regain than other groups. The result will be presented at the conference. The findings of this study not only can help optimize nutrition, cost reduction, and shorter NICU admission of VLBW neonates at the hospital level but eventually contribute to reduced healthcare-associated infections (HAIs) and an improved health economy.

Keywords: very low birth weight neonates, weight gain, parenteral nutrition, glucose, amino acids

Procedia PDF Downloads 62

Authors: Lukas Stiburek, Jana Cesnekova, Josef Houstek, Jiri Zeman

Abstract:

Cellular function depends on mitochondrial function and integrity that is therefore maintained by several classes of proteins possessing chaperone and/or proteolytic activities. In this work, we focused on characterization of LACE1 (lactation elevated 1) function in mitochondrial protein homeostasis maintenance. LACE1 is the human homologue of yeast mitochondrial Afg1 ATPase, a member of SEC18-NSF, PAS1, CDC48-VCP, TBP family. Yeast Afg1 was shown to be involved in mitochondrial complex IV biogenesis, and based on its similarity with CDC48 (p97/VCP) it was suggested to facilitate extraction of polytopic membrane proteins. Here we show that LACE1, which is a mitochondrial integral membrane protein, exists as part of three complexes of approx. 140, 400 and 500 kDa and is essential for maintenance of fused mitochondrial reticulum and lamellar cristae morphology. Using affinity purification of LACE1-FLAG expressed in LACE1 knockdown background we show that the protein physically interacts with mitochondrial inner membrane protease YME1L. We further show that human LACE1 exhibits significant pro-apoptotic activity and that the protein is required for normal function of the mitochondrial respiratory chain. Thus, our work establishes LACE1 as a novel factor with the crucial role in mitochondrial homeostasis maintenance.

Keywords: LACE1, mitochondria, apoptosis, protease

Procedia PDF Downloads 286

Authors: Asif Bilal, Fouzia Tanvir, Sibtain Ahmad

Abstract:

Estrogen receptor alpha, also known as estrogen receptor-1, is highly involved in risk of mammary carcinoma. The objectives of this study were to identify non-synonymous SNPs of estrogen receptor and their association with breast cancer and to identify the chemotherapeutic responses of phytochemicals against it via in-silico study design. For this purpose, different online tools. to identify pathogenic SNPs the tools were SIFT, Polyphen, Polyphen-2, fuNTRp, SNAP2, for finding disease associated SNPs the tools SNP&GO, PhD-SNP, PredictSNP, MAPP, SNAP, MetaSNP, PANTHER, and to check protein stability Mu-Pro, I-Mutant, and CONSURF were used. Post-translational modifications (PTMs) were detected by Musitedeep, Protein secondary structure by SOPMA, protein to protein interaction by STRING, molecular docking by PyRx. Seven SNPs having rsIDs (rs760766066, rs779180038, rs956399300, rs773683317, rs397509428, rs755020320, and rs1131692059) showing mutations on I229T, R243C, Y246H, P336R, Q375H, R394S, and R394H, respectively found to be completely deleterious. The PTMs found were 96 times Glycosylation; 30 times Ubiquitination, a single time Acetylation; and no Hydroxylation and Phosphorylation were found. The protein secondary structure consisted of Alpha helix (Hh) is (28%), Extended strand (Ee) is (21%), Beta turn (Tt) is 7.89% and Random coil (Cc) is (44.11%). Protein-protein interaction analysis revealed that it has strong interaction with Myeloperoxidase, Xanthine dehydrogenase, carboxylesterase 1, Glutathione S-transferase Mu 1, and with estrogen receptors. For molecular docking we used Asiaticoside, Ilekudinuside, Robustoflavone, Irinoticane, Withanolides, and 9-amin0-5 as ligands that extract from phytochemicals and docked with this protein. We found that there was great interaction (from -8.6 to -9.7) of these ligands of phytochemicals at ESR1 wild and two mutants (I229T and R394S). It is concluded that these SNPs found in ESR1 are involved in breast cancer and given phytochemicals are highly helpful against breast cancer as chemotherapeutic agents. Further in vitro and in vivo analysis should be performed to conduct these interactions.

Keywords: breast cancer, ESR1, phytochemicals, molecular docking

Procedia PDF Downloads 45

Authors: Hamza Javar Magnier, Robin Curtis

Abstract:

There have been rapid advances in the upstream processing of protein therapeutics, which has shifted the bottleneck to downstream purification and formulation. Finding liquid formulations with shelf lives of up to two years is increasingly difficult for some of the newer therapeutics, which have been engineered for activity, but their formulations are often viscous, can phase separate, and have a high propensity for irreversible aggregation1. We explore means to develop improved predictive ability from a better understanding of how protein-protein interactions on formulation conditions (pH, ionic strength, buffer type, presence of excipients) and how these impact upon the initial steps in protein self-association and aggregation. In this work, we study the initial steps in the aggregation pathways using a minimal protein model based on square-well potentials and discontinuous molecular dynamics. The effect of model parameters, including range of interaction, stiffness, chain length, and chain sequence, implies that protein models fold according to various pathways. By reducing the range of interactions, the folding- and collapse- transition come together, and follow a single-step folding pathway from the denatured to the native state2. After parameterizing the model interaction-parameters, we developed an understanding of low-temperature conformational properties and fluctuations, and the correlation to the folding transition of proteins in isolation. The model fluctuations increase with temperature. We observe a low-temperature point, below which large fluctuations are frozen out. This implies that fluctuations at low-temperature can be correlated to the folding transition at the melting temperature. Because proteins “breath” at low temperatures, defining a native-state as a single structure with conserved contacts and a fixed three-dimensional structure is misleading. Rather, we introduce a new definition of a native-state ensemble based on our understanding of the core conservation, which takes into account the native fluctuations at low temperatures. This approach permits the study of a large range of length and time scales needed to link the molecular interactions to the macroscopically observed behaviour. In addition, these models studied are parameterized by fitting to experimentally observed protein-protein interactions characterized in terms of osmotic second virial coefficients.

Keywords: protein folding, native-ensemble, conformational fluctuation, aggregation

Procedia PDF Downloads 337

Authors: Oqba Basal, Andras Szabo

Abstract:

Using mineral fertilization is increasing worldwide, as it is claimed to be majorly responsible for achieving high yields; however, the negative impacts of mineral fertilization on soil and environment are becoming more obvious, with alternative methods being more necessary and applicable, especially with the current climatic changes which have imposed serious abiotic stresses, such as drought. An experiment was made during 2017 growing season in Debrecen, Hungary to investigate the effects of inoculation and N fertilization on the seed yield and protein concentration of the soybean (Glycine max (L.) Merr.) cultivar (Panonia Kincse) under three different irrigation regimes: severe drought stress (SD), moderate drought stress (MD) and control with no drought stress (ND). Three N fertilizer rates were applied: no N fertilizer (0 N), 35 kg ha⁻¹ of N fertilizer (35 N) and 105 kg ha⁻¹ of N fertilizer (105 N). Half of the seeds in each treatment was inoculated with Bradyrhizobium japonicum inoculant, and the other half was not inoculated. The results showed significant differences in the seed yield associated with inoculation, irrigation and the interaction between them, whereas there were no significant differences in the seed yield associated with fertilization alone or in interaction with inoculation or irrigation or both. When seeds were inoculated, yield was increased when (35 N) was applied compared to (0 N) but not significantly; however, the high rate of N fertilizer (105 N) reduced the yield to a level even less than (0 N). When seeds were not inoculated, the highest rate of N increased the yield the most compared to the other two N fertilizer rates whenever the drought was present (moderate or severe). Under severe drought stress, inoculation was positively and significantly correlated with yield; however, adding N fertilizer increased the yield of uninoculated plants compared to the inoculated ones, regardless of the rate of N fertilizer. Protein concentration in the seeds was significantly affected by irrigation and by fertilization, but not by inoculation. Protein concentration increased as the N fertilization rate increased, regardless of the inoculation or irrigation treatments; moreover, increasing the N rate reduced the correlation coefficient of protein concentration with the irrigation. It was concluded that adding N fertilizer is not always recommended, especially when seeds are inoculated before being sown; however, it is very important under severe drought stress to sustain yield. Enhanced protein concentrations could be achieved by applying N fertilization, whether the seeds were pre-inoculated or not.

Keywords: drought stress, N fertilization, protein concentration, soybean

Procedia PDF Downloads 129

Authors: Meenakshi Chaudhary, V .S. Randhawa, M. Jais, R. Dutta

Abstract:

Introduction: An outbreak of Multi drug resistant S. Typhi (i.e. resistance to chloramphenicol, ampicillin, and trimethoprim-sulfamethoxazole) occurred in 1990's in India which peaked in 1992-93 and resulted in the change of drug of choice from chloramphenicol to ciprofloxacin for enteric fever. Currently an emergence of Ciprofloxacin susceptible S. Typhi isolates in the region is being reported which appears to be chromosomally mediated. Methodology: Six hundred sixty four strains were randomly selected from the time period between January 2008-December 2011 at the National Salmonella Phage Typing Centre, LHMC, New Delhi. The strains were representative of the north, central and south zones of India. All isolates were subjected to serotyping, biotyping, phage typing and then to antimicrobial susceptibility testing by CLSI disk diffusion (CLSI) technique to Ciprofloxacin, Cefotaxime, Ampicillin, Chloramphenicol, Trimethoprim-Sulfomethoxazole and Tetracycline. Subsequently MIC of the isolates was determined by E-test (AB-Biodisc). Results: More than 80% of the tested strains had intermediate susceptibility to ciprofloxacin. The E test revealed the MIC (Ciprofloxacin) of these strains to be in the range of 0.12 to 0.5 µg/ml. Sixty nine percent of ciprofloxacin intermediate susceptible strains belonged to Phage type E1 and fourteen percent of these were Vi- Negative i.e these could not be typed by the phage typing scheme of Craigie and Yen. All the strains remained susceptible to cefotaxime. Conclusion: Predominant isolation of intermediate susceptible S. Typhi strains from India would alter the recommendations of empiric treatment of enteric fever in the region. Alternative to the low cost ciprofloxacin will have to be sought or increased dosage and/or duration of ciprofloxacin will have to be recommended. The reasons for the trend of increase in percentage of intermediate susceptible S. Typhi strains are not clear but may be attributed partly to the revision of CLSI guidelines in 2013.

Keywords: salmonella typhi, decreased ciprofloxacin susceptibility, ciprofloxacin, minimum inhibitory concentration

Procedia PDF Downloads 294

Authors: Nelson R. Osungbemiro, O. A. Bello-Olusoji, M. Oladipupo

Abstract:

This study was carried out to determine the effects of probiotics Pseudomonas fluorescens on the growth performance, histology examination and immune modulation of African Catfish, (Clarias gariepinus) challenged with Clostridium botulinum. P. fluorescens, and C. botulinum isolates were removed from the gut, gill and skin organs of procured adult samples of Clarias gariepinus from commercial fish farms in Akure, Ondo State, Nigeria. The physical and biochemical tests were performed on the bacterial isolates using standard microbiological techniques for their identification. Antibacterial activity tests on P. fluorescens showed inhibition zone with mean value of 3.7 mm which indicates high level of antagonism. The experimental diets were prepared at different probiotics bacterial concentration comprises of five treatments of different bacterial suspension, including the control (T1), T2 (10³), T3 (10⁵), T4 (10⁷) and T5 (10⁹). Three replicates for each treatment type were prepared. Growth performance and nutrients utilization indices were calculated. The proximate analysis of fish carcass and experimental diet was carried out using standard methods. After feeding for 70 days, haematological values and histological test were done following standard methods; also a subgroup from each experimental treatment was challenged by inoculating Intraperitonieally (I/P) with different concentration of pathogenic C. botulinum. Statistically, there were significant differences (P < 0.05) in the growth performance and nutrient utilization of C. gariepinus. Best weight gain and feed conversion ratio were recorded in fish fed T4 (10⁷) and poorest value obtained in the control. Haematological analyses of C. gariepinus fed the experimental diets indicated that all the fish fed diets with P. fluorescens had marked significantly (p < 0.05) higher White Blood Cell than the control diet. The results of the challenge test showed that fish fed the control diet had the highest mortality rate. Histological examination of the gill, intestine, and liver of fish in this study showed several histopathological alterations in fish fed the control diets compared with those fed the P. fluorescens diets. The study indicated that the optimum level of P. fluorescens required for C. gariepinus growth and white blood cells formation is 10⁷ CFU g⁻¹, while carcass protein deposition required 10⁵ CFU g⁻¹ of P. fluorescens concentration. The study also confirmed P. fluorescens as efficient probiotics that is capable of improving the immune response of C. gariepinus against the attack of a virulent fish pathogen, C. botulinum.

Keywords: Clarias gariepinus, Clostridium botulinum, probiotics, Pseudomonas fluorescens

Procedia PDF Downloads 129

Authors: Moses Ikechukwu Benjamin

Abstract:

The continued increase in methicillin-resistant Staphylococcuspseudintermedius (MRSP) among dogs and the zoonotic transmission event of MRSP from dogs to humans threaten veterinary medicine and public health. The cardinal objective of this study was to determine the antibiogram and frequency of toxingenes in MRSP obtained from shelter dogs with skin infections and dog owners in Abakaliki, Eastern Nigeria. Skinswabs from 61 shelter dogs with skin infections and 33 nasal swabs from dog owners were processed and analyzed using standard microbiological techniques. Susceptibility to antibiotics was determined by Kirby Bauer disc diffusion technique. The screening for Seccanine, lukD, siet, and exitoxin genes was carried out by PCR. A total of 23 (37.7 %) and 1 (3 %) MRSP strains were obtained from shelter dogs and dog owners, respectively. Generally, isolates exhibited high resistance to amoxicillin-clavulanic acid, ceftazidime, and cefepime (100 % - 66.7 %) but were very susceptible (100 % - 70.7 %) to chloramphenicol and doripenem. The only isolate from dog owners harbouredseccanine, lukD, and siet toxin genes while solatesfrom shelter dogs harbouredseccanine16 (69.6 %), lukD 17 (73.9 %), siet 20 (87 %), and exi1 (4.4 %) toxin genes. Isolates were generally observed to be more resistant than other reports from the literature. Interesting, there was a similarity in the resistance antibiotypes and frequency of toxin genes harboured by MRSP isolates between shelter dogs with skin infections and their owner in a sampled household, thus suggesting a likely zoonotic transmission event. This report of the occurrence of MRSP and high frequency of toxin genes (Seccanine,lukD, and siet) in shelter dogs and dog owners represent a major challenge, especially in terms of antibiotic therapy, and is a serious concern for both animal and public health.

Keywords: methicillin-resistant S. pseudintermedius, zoonotic transmission, antibiotic resistance, companion dogs, toxin genes

Procedia PDF Downloads 138

Authors: Sarita Puri, Tapan K. Chaudhuri

Abstract:

Chaperonin GroEL is a tetradecameric Escherichia coli protein having identical subunits of 57 kDa. The elucidation of thermodynamic parameters related to stability for the native GroEL is not feasible as it undergoes irreversible unfolding because of its large size (800kDa) and multimeric nature. Nevertheless, it is important to determine the thermodynamic stability parameters for the highly stable GroEL protein as it helps in folding and holding of many substrate proteins during many cellular stresses. Properly folded monomers work as building-block for the formation of native tetradecameric GroEL. Spontaneous refolding behavior of monomeric GroEL makes it suitable for protein-denaturant interactions and thermodynamic stability based studies. The urea mediated unfolding is a three state process which means there is the formation of one intermediate state along with native and unfolded states. The heat mediated denaturation is a two-state process. The unfolding process is reversible as observed by the spontaneous refolding of denatured protein in both urea and head mediated refolding processes. Analysis of folding/unfolding data provides a measure of various thermodynamic stability parameters for the monomeric GroEL. The proposed mechanism of unfolding of monomeric GroEL is a three state process which involves formation of one stable intermediate having folded apical domain and unfolded equatorial, intermediate domains. Research in progress is to demonstrate the importance of specific residues in stability and oligomerization of GroEL protein. Several mutant versions of GroEL are under investigation to resolve the above mentioned issue.

Keywords: equilibrium unfolding, monomeric GroEl, spontaneous refolding, thermodynamic stability

Procedia PDF Downloads 254

Authors: Saba Belgheisi

Abstract:

Food packaging, is an important discipline in the area of food technology, concerns preservation and protection of foods. The objective of this research was to determine of the effect of whey protein based edible coating on the moisture loss and sensory attributes of fresh mutton after 0, 1, 3 and 5 days at 5° C. The moisture content, moisture loss and sensory attributes (juiciness, color and odor) of the coated and uncoated samples were analyzed. The results showed that, moisture content, moisture loss, juiciness and color of the coated and uncoated samples have significant differences (p < 0.05) at the intervals of 0 to 1 and 1 to 3 days of storage. But no significant difference was observed at interval time 3 to 5 days of storage (p > 0.05). Also, there was no significant differences in the odor values of the coated and uncoated samples (p > 0.05). Therefore, the coated samples had consistently more moisture, juiciness and colored values than uncoated samples after 3 days at 5° C. So, whey protein edible coating could enhance product presentation and eliminate the need for placing absorbent pads at the bottom of the trays.

Keywords: coating, whey protein, mutton, moisture, sensory

Procedia PDF Downloads 442

Authors: Yazeed Al-Sheikh

Abstract:

Pregnancy represents a risk factor in the occurrence of vulvovaginal candidiasis. To investigate the prevalence rate of vaginal carriage of Candida species in Saudi pregnant and non-pregnant women, high vaginal swab (HVS) specimens (707) were examined by direct microscopy (10% KOH and Giemsa staining) and parallel cultured on Sabouraud Dextrose Agar (SDA) as well as on “CHROM agar Candida” medium. As expected, Candida-positive cultures were frequently observed in pregnant-test group (24%) than in non-pregnant group (17%). The frequency of culture positive was correlated to pregnancy (P=0.047), parity (P=0.001), use of contraceptive (P=0.146), or antibiotics (P=0.128), and diabetic-patients (P < 0.0001). Out of 707 HVS examined specimens, 157 specimens were yeast-positive culture (22%) on Sabouraud Dextrose Agar or “CHROM agar Candida”. In comparison, the sensitivities of the direct 10% KOH and the Giemsa stain microscopic examination methods were 84% (132/157) and 95% (149/157) respectively but both with 100% specificity. As for the identity of recovered 157 yeast isolates, based on API 20C biotype carbohydrate assimilation, germ tube and chlamydospore formation, C. albicansand C. glabrata constitute 80.3 and 12.7% respectively. Rates of C. tropicalis, C. kefyr, C. famata or C. utilis were 2.6, 1.3, and 0.6% respectively. Sachromyces cerevisiae and Rhodotorula mucilaginosa yeasts were also encountered at a frequency of 1.3 and 0.6% respectively. Finally, among all recovered 157 yeast-isolates, strains resistant to ketoconazole were not detected, whereas 5% of the C. albicans and as high as 55% of the non-albicans yeast isolates (majority C. glabrata) showed resistance to fluconazole. Our findings may prove helpful for continuous determination of the existing vaginal candidiasis causative species during pregnancy, its lab-diagnosis and/or control and possible measures to minimize the incidence of the disease-associated pre-term delivery.

Keywords: vaginal candidiasis, Candida spp., pregnancy, risk factors, API 20C-yeast biotypes, giemsa stain, antifungal agents

Procedia PDF Downloads 221

Authors: Nwafia Ifeyinwa Nkeiruka, Nwafia Walter Chukwuma

Abstract:

Since the early 1980’s fungi have emerged as major causes of human diseases, especially among immunocompromised. The most commonly isolated yeast is Candida albicans and constitutes the 4th most common nosocomial BSI in humans. It is progressive and cumulative and become more complex over time.It can even lead to leaky gut syndrome that causes food and environmental allergies. It is worthy of note that all the available data on oral Candida risk factors in humans were documented essentially using data from studies conducted in other areas, hence there is need for comparative and complementary information from the South eastern part of Nigeria. Method: 200 subjects of all age groups of both sexes were randomly examined,by swabbing their palatine mucosa and dorsal tongue with sterile cotton wool,then cultured into Sabouraud dextrose agar plates supplemented with antibiotics and incubated aerobically at 37 degree for 48 hrs. Identification of Candida albicans was done by germ tubes tests, chlamydospores production on cornmeal agar supplemented with 1% Tween 80.Sugar and nitrogen assimilation test using API 20C Auxanogram and potassium nitrate agar. Results: Out of 30 samples that were positive for candida, 15 (50%) were candida albicans. Using the anova test (P < 0.05) this variation is significant (P = 0016). followed by C. dublinensis 3 (13%), C. tropicalis 3 (10%), C. pseudotropicalis 3 (10%), C, glabrata 2 (7%), C. parapsilosis 2 (7%) and lastly C. krusei 1 (3%).However, 53% of the patients were female while 47% were male. Among the HIV positive isolates.67% were HIV isolates not on drugs while 33% positives isolates were on drugs and the percentages of candida species in these patients were as follows C. albicans were 45% followed by C. glabrata and C.tropicalis which were 17% each, C.parapsilosis, C.dubliensis and C.pseudotropicalis were all 8% each. Conclusion: Oral Candidiasis is a marker of systemic diseases and in some cases, it may be the first clinical presentation. There is need for more intensive clinical and laboratory monitoring and possible early intervention to prevent the reoccurrence and resistance to treatment.

Keywords: oral cavity, Candida species, oral Candidiasis, risk factors

Procedia PDF Downloads 339

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

Abstract:

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

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

Procedia PDF Downloads 158

Authors: Sodamade A., Bolaji K. A.

Abstract:

Problems associated with protein malnutrition are still prevalent in third-world countries, leading to the constant search for plants that could serve as nutrients and medicinal purposes. Huckleberry is one of the plants that has been proven useful locally in the treatment of numerous ailments and diseases. A fresh sample of the plant (Vaccinium pavifolium) was collected from a vegetable garden situated near the Erelu dam of the Emmanuel Alayande College of Education Campus, Oyo. The sample was authenticated at the Forestry Research Institute of Nigeria (FRIN) Ibadan. The leaves of the plant were plucked and processed for leaf protein concentrates before proximate composition, mineral analysis phytochemical and antimicrobial properties were determined using a standard method of analysis. The results of proximate constituents showed; moisture content; 9.89±0.051g/100g, Ash; 3.23±0.12g/100g, crude fat; 3.96±0.11g/100g and 61.27±0.56g/100g of Nitrogen free extractive. The mineral analysis of the sample showed; Mg; 0.081±0.00mg/100g, Ca; 42.30±0.05mg/100g, Na; 27.57±0.09mg/100g, K; 6.81±0.01mg/100g, P; 8.90±0.03mg/100g, Fe; 0.51±0.00mg/100g, Zn; 0.021±0.00mg/100g, Cd; 0.04±0.04mg/100g, Pb; 0.002±0.00mg/100g, Cr; 0.041±0.00mg/100g Cadmium and Mercury were not detected in the sample. The result of phytochemical analysis of leaf protein concentrates of the Huckleberry showed the presence of Alkaloid, Saponin, Flavonoid, Tanin, Coumarin, Steroids, Terpenoids, Cardiac glycosides, Glycosides, Quinones, Anthocyanin, phytosterols, and phenols. Ethanolic extracts of the Vaccinium parvifolium L. leaf protein concentrates showed that it contains bioactive compounds that are capable of combating the following microorganisms; Staphylococcus aureus, Streptococcus pyogenes, Streptococcus faecalis, Pseudomonas aeruginosa, Klebisialae pneumonia and Proteus mirabilis. The results of the analysis of Vaccinium parvifolium L. leaf protein concentrates showed that the sample contains valuable nutrient and mineral constituents, and phytochemical compounds that could make the sample useful for medicinal activities.

Keywords: leaf protein concentrates, vaccinium parvifolium, nutritional characteristics, mineral composition, antimicrobial activity

Procedia PDF Downloads 49

Authors: Hemalatha Ganapathyswamy, Thirukkumar Subramani

Abstract:

Celiac disease is an autoimmune response to dietary wheat gluten. Gluten is the main structure forming protein in bread and hence developing gluten-free bread is a technological challenge. The study aims at using nonwheat flours like barnyard millet and lentil flour to replace wheat in bread formulations. Other characteristics of these grains, such as high protein, soluble fiber, mineral content and bioactive components make them attractive alternatives to traditional gluten-free ingredients in the production of high protein, gluten-free bread. The composite flour formulations for the development of gluten-free bread were optimized using lentil flour (50 to 70 g), barnyard millet flour (0 to 30 g) and corn flour (0 to 30 g) by means of response surface methodology with various independent variables for physical, sensorial and nutritional characteristics. The optimized composite flour which had a desirability value of 0.517, included lentil flour –62.94 g, barnyard millet flour– 24.34 g and corn flour– 12.72 g with overall acceptability score 8.00/9.00. The optimized gluten-free bread formulation had high protein (14.99g/100g) and fiber (1.95g/100g) content. The glycemic index of the gluten-free bread was 54.58 rendering it as low glycemic which enhances the functional benefit of the gluten-free bread. Since the standardised gluten-free bread from barnyard millet and lentil flour are high protein, and gluten-free with low glycemic index, the product would serve as an ideal therapeutic food in the management of both celiac disease and diabetes mellitus with better nutritional value.

Keywords: gluten free bread, lentil, low glycemic index, response surface methodology

Procedia PDF Downloads 167

Authors: M. H. Nahaisi, N. M. Almaroum

Abstract:

Laban is a Libyan traditional fermented milk product. This lactic fermentation has been known in many cities of Libya long time ago as stable, nutritious, refreshing drink especially during the summer. 16 naturally fermented milk samples were collected from different cities located in North West of Libya. The average pH, titratable acidity, fat and total solids were 4.16, 0.73%, 1.54% and 8.12 % respectively. Coliform, yeast and mold counts were 21×10⁴, 39×10⁴ and 41 ×10³ cfu/ ml. respectively. The average Lactococcus, Streptococcus, Mesophilic Lactobacillus / Leuconostoc and Thermophilic Lactobacillus counts were 99 ×10⁷, 96 ×10⁷, 93 ×10⁷ and 15 ×10⁷ cfu / ml. respectively. A total of one hundred forty two lactic acid bacteria (LAB) isolates were identified to the genus level as Lactobacillus (48.59%), Lactococcus (43.66%), Streptococcus (4.93%) and Leuconostoc (2.82%). Sugar fermentation tests have revealed that the most frequently Lactobacillus species was found to be Lactobacillus delbrueckii ssp. lactis (62.32%) followed by Lactobacillus plantarum (31.88%). Furthermore, other selected LAB isolates were identified by API 50 CH test as Lactococcus lactis ssp. lactics, Lactobacillus pentosus, Lactobacillus brevis and Leuconostoc mesenteroides ssp. cremoris.

Keywords: traditional fermented milk, laban, lactococcus, streptococcus, mesophilic lactobacillus, thermophilic lactobacillus counts

Procedia PDF Downloads 351

Authors: Harison Masih, Jyotsna Kiran Peter, Sundara Singh, Geetha Singh

Abstract:

The present investigation deals with diversity of dermatophytes and keratinophilic fungi from different tourist spots such as Agra Fort, Akbar tomb, It-Mat-Ud-Daulah, Mariam tomb, Radha Swami Bagh, and Taj Mahal of Agra City. These fungi are medically important which causes various infections and diseases in humans and animals. The main reservoir of these pathogens are the keratinous substances that increases due to birds and animal activities in the vicinity of monuments, where thousands (5413266) annual visitors from all over the world are visiting. The soil samples were subjected to isolate the pathogenic fungi through bait technique (buffalo skin, chicken feathers, human hair and goat tail hair). Baits were spread over the soil samples and incubated at room temperature for 30-35 days and pure culture isolates were maintained in SDA medium, stored at 4°C. Highest number of visitors were (3906453) from Taj Mahal, minimum 10785 at Mariam tomb annually, the total 271 isolates were encountered from soil samples out of these 18 genera and 38 species were found in different season. Highest incidence was 4.79% frequency shown by Chrysosporium keratinophilum while least 738% frequency occurrence by Trichophyton simii in soil samples. From the present study it was concluded that the incidence of pathogenic fungal isolates were the common in tourists soil that are etiological agents of superficial mycosis. Thus, both human and animal activity seemed to play an important role in occurrence and distribution of keratinophilic and related dermatophytes at various tourist places of Agra city.

Keywords: dermatophytic fungal diversity, bait technique, visitors at tourist spots, human and animal activities, soil samples

Procedia PDF Downloads 455

Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

Abstract:

Klebsiella pneumoniae causes a wide range of infections, including urinary tract infections, sepsis, bacteremia, pneumonia, and liver abscesses. The emergence of multi-drug resistance in this bacterium led to a major setback for clinical management. WHO also endorsed a need for finding alternative therapy to antibiotics for the treatment of these infections. Development of vaccines and passive antibody therapy has been proven as a potent alternative to antibiotics in the case of MDR, XDR, and PDR Klebsiella infections. Siderophore receptors have been demonstrated to be overexpressed for the internalization of iron siderophore complexes during infections in most Gram-negative bacteria. For the present study, immune response to siderophore receptors to establish this protein as a potential immunogen for the development of therapeutic leads was explored. Clinical strains of Klebsiella pneumoniae were grown in iron-deficient conditions, and the iron-regulated outer membrane proteins were extracted and characterized through mass spectrometry for specific identification. The gene for identified protein was cloned in pET- 28a vector and expressed in E. coli. The native protein and the recombinant protein were isolated and purified and used as antigens for the generation of immune response in BALB/c mice. The native protein of Klebsiella pneumoniae grown in iron-deficient conditions was identified as FepA (Ferrienterobactin receptor) and other siderophore receptors. This 80 kDa protein generated an immune response in BALB/c mice. The antiserum from mice after subsequent booster doses was collected and showed binding with FepA protein in western blot and phagocytic uptake of the K. pneumoniae in the presence antiserum from immunized mice also observed from the animal studies after bacterial challenge post immunisation in mice have shown bacterial clearance. The antiserum from mice showed binding and clearance of the Klebsiella pneumoniae bacteria in vitro and in vivo. These antigens used for generating an active immune response in mice can further be used for therapeutic monoclonal antibody development against Klebsiella pneumoniae infections.

Keywords: antiserum, FepA, Klebsiella pneumoniae, multi drug resistance, siderophore receptor

Procedia PDF Downloads 77