Search results for: milk proteins
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1443

Search results for: milk proteins

543 Mycoplasmas and Pathogenesis in Preventive Medicine

Authors: Narin Salehiyan

Abstract:

The later sequencing of the complete genomes of Mycoplasma genitalium and M. pneumoniae has pulled in significant consideration to the atomic science of mycoplasmas, the littlest self-replicating living beings. It shows up that we are presently much closer to the objective of defining, in atomic terms, the complete apparatus of a self-replicating cell. Comparative genomics based on comparison of the genomic cosmetics of mycoplasmal genomes with those of other microbes, has opened better approaches of looking at the developmental history of the mycoplasmas. There's presently strong hereditary bolster for the speculation that mycoplasmas have advanced as a department of gram-positive microbes by a handle of reductive advancement. Amid this prepare, the mycoplasmas misplaced significant parcels of their ancestors’ chromosomes but held the qualities basic for life. In this way, the mycoplasmal genomes carry a tall rate of preserved qualities, incredibly encouraging quality comment. The critical genome compaction that happened in mycoplasmas was made conceivable by receiving a parasitic mode of life. The supply of supplements from their has clearly empowered mycoplasmas to lose, amid advancement, the qualities for numerous assimilative forms. Amid their advancement and adjustment to a parasitic mode of life, the mycoplasmas have created different hereditary frameworks giving a profoundly plastic set of variable surface proteins to avoid the have safe framework.

Keywords: mycoplasma, plasma, pathogen, genome

Procedia PDF Downloads 40
542 Preventive and Attenuative Effect of Vitamin E on Selenite-induced Cataract in Rat

Authors: Seyedeh Zeinab Peighambarzadeh, Mehdi Tavana

Abstract:

Cataract is the most common cause of blindness worldwide and its incidence will increase as the World’s population ages. Even in modern ophthalmology, there is no effective medical treatment for cataract except surgery. Development of a drug which could prevent or delay the onset of cataract will lessen this burden and reduce the number of blind patients waiting for cataract surgery. This study was undertaken to evaluate the protective effect of vitamin E on Selenite-induced Cataract in Sprague-dawely rats. Cataracts were induced in rats by administration of sodium selenite. On postpartum day ten, in group I, saline was injected subcutaneously. Group II rat pups received subcutaneous injection of vitamin E (60mg/kg B.W.) at day 8 postpartum and every other day thereafter. Group III and IV rat pups received a subcutaneous injection of sodium selenite (13mg/kg B.W.) at day 10 postpartum. Group IV also received subcutaneous injection of vitamin E (60mg/kg B.W.) at day 8 postpartum and every other day thereafter. The development of cataract in rats was assessed clinically by slit-lamp biomicroscope from day 14 up to postpartum day 28. After sacrifice, extricated pup lenses were analyzed for total and soluble protein concentrations and eletrophoretic pattern (SDS-PAGE). There was no opacification of lens in Group I and II. There was mature cataract in 95% of Group III. In group IV, 55% of rats developed sub capsular or cortical cataract. Cataractous and biochemical changes of the crystalline lens proteins due to selenite can be retard or prevented by vitamin E.

Keywords: preventive effect, selenite-induced cataract, vitamin E, rat

Procedia PDF Downloads 359
541 Breastfeeding Experiences of Nutritionist who are Mothers in Quito- Ecuador

Authors: Maria Jose Mendoza Gordillo

Abstract:

Introduction: Research regarding breastfeeding is devoted to how essential breastfeeding is to guarantee wellbeing for the mother and the baby from a medical standpoint relegating the cultural, material and social barriers for breastfeeding. Consequently, worldwide breastfeeding rates are low, and Ecuador is not the exception, especially among working mothers. Worldwide, health care providers have low rates of breastfeeding due to several barriers to lactation, such as the work schedule, a lack of private places for pumping while at work, and negative emotions. Goals and Methods: This study aimed to explore how do Ecuadorian women embrace their identities as nutritionists and mothers within their breastfeeding experience. The primary data come from 20 synchronous semi-structured interviews, which follow a topic guide. The interviews were recorded and transcribed verbatim. The data analysis followed the Phronetic Iterative Approach. Results: Women shifted the preconceived idea of the ideal breastfeeding that came from the medicalized discourse of breastfeeding, and that was constructed in their training as nutritionists. Although these women believe that breast milk and breastfeeding is the best way to feed a baby, the internalized ideal of breastfeeding shifted through the experience of motherhood. When these women developed their identity as mothers, they understood that the ideal breastfeeding is different from the medicalized discourse. Although they have that clash between the ideal and the external reality, they continued breastfeeding their babies and those experiences made them improve their professional practice. Conclusions: The narratives that women shared illustrate how complex it was to manage the different roles and identities that they wanted to fulfill to keep their identity of a good mother who breastfeeds her baby and, at the same time, a good healthcare provider identity. The process of breastfeeding for this group of women who are mothers and healthcare professionals appears to be a unique relational and identity negotiation process.

Keywords: breastfeeding, identity, nutritionist, qualitative

Procedia PDF Downloads 157
540 In-Vivo Association of Multivalent 11 Zinc Fingers Transcriptional Factors CTCF and Boris to YB-1 in Multiforme Glioma-RGBM Cell Line

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

Abstract:

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

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

Procedia PDF Downloads 246
539 Sequence Analysis of the Effect of HPV-16 E1 Variation on Cervical Carcinogenesis

Authors: Fern Baedyananda, Arkom Chaiwongkot, Somchai Niruthisard, Nakarin Kitkumthorn, Parvapan Bhattarakosol

Abstract:

High-risk human papillomavirus (HPV) infections cause transformation of the host cells by down-regulating and inhibiting host regulatory proteins such as p53 and pRb by overexpressing the viral oncoproteins E6 and E7. However, the E1 protein which is the only enzyme encoded by HPV has also been shown to cause DNA instability leading to the integration of the virus into the host genome and triggering carcinogenic events. A 63bp duplication in the E1 helicase region has been detected in European patients. However, the clinical prognosis of these patients is still controversial. This study was performed to determine the presence of the HPV-16 E1 63bp duplication in patient cervical samples in Thai women and determine the sequence of the variant in the Thai population. Detection of the HPV-16 E1 duplication in the helicase region was performed in 90 patient cell samples across normal, cervical intraepithelial neoplasia I-III, and squamous cervical carcinoma stages by PCR. The PCR products were purified and sequenced to determine the presence of duplication variants.The variant form was found in 10% of all CIN 1 patients. In this study, the presence of the 63 bp duplication variant in the Thai population was found to be present and was further characterized. Interestingly, all samples that exhibited the variant form of HPV-16 E1 were classified as CIN I. Presence of the variant, constricted to mild dysplasia signifies the importance of HPV-16 E1 in carcinogenesis.

Keywords: carcinogenesis, cervical cancer, human papillomavirus, HPV-16 E1

Procedia PDF Downloads 216
538 Solid State Fermentation Process Development for Trichoderma asperellum Using Inert Support in a Fixed Bed Fermenter

Authors: Mauricio Cruz, Andrés Díaz García, Martha Isabel Gómez, Juan Carlos Serrato Bermúdez

Abstract:

The disadvantages of using natural substrates in SSF processes have been well recognized and mainly are associated to gradual decomposition of the substrate, formation of agglomerates and decrease of porosity bed generating limitations in the mass and heat transfer. Additionally, in several cases, materials with a high agricultural value such as sour milk, beets, rice, beans and corn have been used. Thus, the use of economic inert supports (natural or synthetic) in combination with a nutrient suspension for the production of biocontrol microorganisms is a good alternative in SSF processes, but requires further studies in the fields of modeling and optimization. Therefore, the aim of this work is to compare the performance of two inert supports, a synthetic (polyurethane foam) and a natural one (rice husk), identifying the factors that have the major effects on the productivity of T. asperellum Th204 and the maximum specific growth rate in a PROPHYTA L05® fixed bed bioreactor. For this, the six factors C:N ratio, temperature, inoculation rate, bed height, air moisture content and airflow were evaluated using a fractional design. The factors C:N and air flow were identified as significant on the productivity (expressed as conidia/dry substrate•h). The polyurethane foam showed higher maximum specific growth rate (0.1631 h-1) and productivities of 3.89 x107 conidia/dry substrate•h compared to rice husk (2.83x106) and natural substrate based on rice (8.87x106) used as control. Finally, a quadratic model was generated and validated, obtaining productivities higher than 3.0x107 conidia/dry substrate•h with air flow at 0.9 m3/h and C:N ratio at 18.1.

Keywords: bioprocess, scale up, fractional design, C:N ratio, air flow

Procedia PDF Downloads 491
537 Cardiac Hypertrophy in Diabetes; The Role of Factor Forkhead Box Class O-Regulation by O-GlcNAcylation

Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian

Abstract:

Cardiac hypertrophy arises in response to persistent increases in hemodynamic loads. In comparison, diabetic cardiomyopathy is defined by an abnormal myocardial changes without other cardiac-related risk factors. Pathological cardiac hypertrophy and myocardial remodeling are hallmarks of cardiovascular diseases and are risk factors for heart failure. The transcription factor forkhead box class O (FOXOs) can protect heart tissue by hostile oxidative stress and stimulating apoptosis and autophagy. FOXO proteins, as sensitive elements and mediators in response to environmental changes, have been revealed to prevent and inverse cardiac hypertrophy. FOXOs are inhibited by insulin and are critical mediators of insulin action. Insulin deficiency and uncontrolled diabetes lead to a catabolic state. FOXO1 acts downstream of the insulin-dependent pathways, which are dysregulated in diabetes. It regulates cardiomyocyte hypertrophy downstream of IGF1R/PI3K/Akt activation, which are critical regulators of cardiac hypertrophy. The complex network of signaling pathways comprising insulin/IGF-1 signaling, AMPK, JNK, and Sirtuins regulate the development of cardiovascular dysfunction by modulating the activity of FOXOs. Insulin receptors and IGF1R act via the PI3k/Akt and the MAPK/ERK pathways. Activation of Akt in response to insulin or IGF-1 induces phosphorylation of FOXOs. Increased protein synthesis induced by activation of the IGF-I/Akt/mTOR signaling pathway leads to hypertrophy. This pathway and the myostatin/Smad pathway are potent negative muscle development regulators. In cardiac muscle, insulin receptor substrates (IRS)-1 or IRS-2 activates the Akt signaling pathway and inactivate FOXO1. Under metabolic stress, p38 MAPK promotes degradation of IRS-1 and IRS-2 in cardiac myocytes and activates FOXO1, leading to cardiomyopathy. Sirt1 and FOXO1 interaction play an essential role in starvation-induced autophagy in cardiac metabolism. Inhibition of Angiotensin-II induced cardiomyocyte hypertrophy is associated with reduced FOXO1 acetylation and activation of Sirt1. The NF-κB, ERK, and FOXOs are de-acetylated by SIRT1. De-acetylation of FOXO1 induces the expression of genes involved in autophagy and stimulates autophagy flux. Therefore, under metabolic stress, FOXO1 can cause diabetic cardiomyopathy. The overexpression of FOXO1 leads to decreased cardiomyocyte size and suppresses cardiac hypertrophy through inhibition of the calcineurin–NFAT pathway. Diabetes mellitus is associated with elevation of O-GlcNAcylation. Some of its binding partners regulate the substrate selectivity of O-GlcNAc transferase (OGT). O-GlcNAcylation of essential contractile proteins may inhibit protein-protein interactions, reduce calcium sensitivity, and modulate contractile function. Uridine diphosphate (UDP)-GlcNAc is the obligatory substrate of OGT, which catalyzes a reversible post-translational protein modification. The increase of O-GlcNAcylation is accompanied by impaired cardiac hypertrophy in diabetic hearts. Inhibition of O-GlcNAcylation blocks activation of ERK1/2 and hypertrophic growth. O-GlcNAc modification on NFAT is required for its translocation from the cytosol to the nucleus, where NFAT stimulates the transcription of various hypertrophic genes. Inhibition of O-GlcNAcylation dampens NFAT-induced cardiac hypertrophic growth. Transcriptional activity of FOXO1 is enriched by improved O-GlcNAcylation upon high glucose stimulation or OGT overexpression. In diabetic conditions, the modification of FOXO1 by O-GlcNAc is promoted in cardiac troponin I and myosin light chain 2. Therefore targeting O-GlcNAcylation represents a potential therapeutic option to prevent hypertrophy in the diabetic heart.

Keywords: diabetes, cardiac hypertrophy, O-GlcNAcylation, FOXO1, Akt, PI3K, AMPK, insulin

Procedia PDF Downloads 92
536 Development of Cost-Effective Protocol for Preparation of Dehydrated Paneer (Indian Cottage Cheese) Using Freeze Drying

Authors: Sadhana Sharma, P. K. Nema, Siddhartha Singha

Abstract:

Paneer or Indian cottage cheese is an acid and heat coagulated milk product, highly perishable because of high moisture (58-60 %). Typically paneer is marble to light creamy white in appearance. A good paneer should have cohesive body with slight sponginess or springiness. The texture must be smooth and velvety with close-knit compactness. It should have pleasing mild acidic, slightly sweet and nutty flavour. Consumers today demand simple to prepare, convenient, healthy and natural foods. Dehydrated paneer finds numerous ways to be used. It can be used in curry preparation similar to paneer-in-curry, a delicacy in Indian cuisine. It may be added to granola/ trail mix yielding a high energy snack. If grounded to a powder, it may be used as a cheesy spice mix or used as popcorn seasoning. Dried paneer powder may be added to pizza dough or to a white sauce to turn it into a paneer sauce. Drying of such food hydrogels by conventional methods is associated with several undesirable characteristics including case hardening, longer drying time, poor rehydration ability and fat loss during drying. The present study focuses on developing cost-effective protocol for freeze-drying of paneer. The dehydrated product would be shelf-stable and can be rehydrated to its original state having flavor and texture comparable to the fresh form. Moreover, the final product after rehydration would be more fresh and softer than its frozen counterparts. The developed product would be shelf-stable at room temperature without any addition of preservatives.

Keywords: color, freeze-drying, paneer, texture

Procedia PDF Downloads 143
535 Resveratrol Ameliorates Benzo(a)Pyrene Induced Testicular Dysfunction and Apoptosis: Involvement of p38 MAPK/ATF2/iNOS Signaling

Authors: Kuladip Jana, Bhaswati Banerjee, Parimal C. Sen

Abstract:

Benzo(a)pyrene [B(a)P] is an environmental toxicant present mostly in cigarette smoke and car exhaust, is an aryl hydrocarbon receptor (AhR) ligand that exerts its toxic effects on both male and female reproductive systems along with carcinogenesis in skin, prostate, ovary, lung and mammary glands. Our study was focused on elucidating the molecular mechanism of B(a)P induced male reproductive toxicity and its prevention with phytochemical like resveratrol. In this study, the effect of B(a)P at different doses (0.1, 0.25, 0.5, 1 and 5 mg /kg body weight) was studied on male reproductive system of Wistar rat. A significant decrease in cauda epididymal sperm count and motility along with the presence of sperm head abnormalities and altered epididymal and testicular histology were documented following B(a)P treatment. B(a)P treatment resulted apoptotic sperm cells as observed by TUNEL and Annexin V-PI assay with increased Reactive Oxygen Species (ROS), altered sperm mitochondrial membrane potential (ΔΨm) with a simultaneous decrease in the activity of antioxidant enzymes and GSH status. TUNEL positive apoptotic cells also observed in testis as well as isolated germ and Leydig cells following B(a)P exposure. Western Blot analysis revealed the activation of p38 mitogen activated protein kinase (p38MAPK), cytosolic translocation of cytochrome-c, upregulation of Bax and inducible nitric oxide synthase (iNOS) with cleavage of poly ADP ribose polymerase (PARP) and down regulation of BCl2 in testis upon B(a)P treatment. The protein and mRNA levels of testicular key steroidogenesis regulatory proteins like steroidogenic acute regulatory protein (StAR), cytochrome P450 IIA1 (CYPIIA1), 3β hydroxy steroid dehydrogenase (3β HSD), 17β hydroxy steroid dehydrogenase (17β HSD) showed a significant decrease in a dose dependent manner while an increase in the expression of cytochrome P450 1A1 (CYP1A1), Aryl hydrocarbon Receptor (AhR), active caspase- 9 and caspase- 3 following B(a)P exposure. We conclude that exposure of benzo(a)pyrene caused testicular gamatogenic and steroidogenic disorders by induction of oxidative stress, inhibition of StAR and other steroidogenic enzymes along with activation of p38MAPK and initiated caspase-3 mediated germ and Leydig cell apoptosis. Next we investigated the role of resveratrol on B(a)P induced male reproductive toxicity. Our study highlighted that resveratrol co-treatment with B(a)P maintained testicular redox potential, increased serum testosterone level and prevented steroidogenic dysfunction with enhanced expression of major testicular steroidogenic proteins (CYPIIA1, StAR, 3β HSD,17β HSD) relative to treatment with B(a)P only. Resveratrol suppressed B(a)P-induced testicular activation of p38 MAPK, ATF2, iNOS and ROS production; cytosolic translocation of Cytochome c and Caspase 3 activation thereby prevented oxidative stress of testis and inhibited apoptosis. Resveratrol co-treatment also decreased B(a)P-induced AhR protein level, its nuclear translocation and subsequent CYP1A1 promoter activation, thereby decreased protein and mRNA levels of testicular cytochrome P4501A1 (CYP1A1) and prevented BPDE-DNA adduct formation. Our findings cumulatively suggest that resveratrol prevents activation of B(a)P by modulating the transcriptional regulation of CYP1A1 and acting as an antioxidant thus prevents B(a)P-induced oxidative stress and testicular apoptosis.

Keywords: benzo(a)pyrene, resveratrol, testis, apoptosis, cytochrome P450 1A1 (CYP1A1), aryl hydrocarbon receptor (AhR), p38 MAPK/ATF2/iNOS

Procedia PDF Downloads 213
534 Effect of Oxytocin on Cytosolic Calcium Concentration of Alpha and Beta Cells in Pancreas

Authors: Rauza Sukma Rita, Katsuya Dezaki, Yuko Maejima, Toshihiko Yada

Abstract:

Oxytocin is a nine-amino acid peptide synthesized in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus. Oxytocin promotes contraction of the uterus during birth and milk ejection during breast feeding. Although oxytocin receptors are found predominantly in the breasts and uterus of females, many tissues and organs express oxytocin receptors, including the pituitary, heart, kidney, thymus, vascular endothelium, adipocytes, osteoblasts, adrenal gland, pancreatic islets, and many cell lines. On the other hand, in pancreatic islets, oxytocin receptors are expressed in both α-cells and β-cells with stronger expression in α- cells. However, to our knowledge there are no reports yet about the effect of oxytocin on cytosolic calcium reaction on α and β-cell. This study aims to investigate the effect of oxytocin on α-cells and β-cells and its oscillation pattern. Islet of Langerhans from wild type mice were isolated by collagenase digestion. Isolated and dissociated single cells either α-cells or β-cells on coverslips were mounted in an open chamber and superfused in HKRB. Cytosolic concentration ([Ca2+]i) in single cells were measured by fura-2 microfluorimetry. After measurement of [Ca2+]i, α-cells were identified by subsequent immunocytochemical staining using an anti-glucagon antiserum. In β-cells, the [Ca2+]i increase in response to oxytocin was observed only under 8.3 mM glucose condition, whereas in α-cells, [Ca2+]i an increase induced by oxytocin was observed in both 2.8 mM and 8.3 mM glucose. The oscillation incidence was induced more frequently in β-cells compared to α-cells. In conclusion, the present study demonstrated that oxytocin directly interacts with both α-cells and β-cells and induces increase of [Ca2+]i and its specific patterns.

Keywords: α-cells, β-cells, cytosolic calcium concentration, oscillation, oxytocin

Procedia PDF Downloads 170
533 Rumination Time and Reticuloruminal Temperature around Calving in Eutocic and Dystocic Dairy Cows

Authors: Levente Kovács, Fruzsina Luca Kézér, Ottó Szenci

Abstract:

Prediction of the onset of calving and recognizing difficulties at calving has great importance in decreasing neonatal losses and reducing the risk of health problems in the early postpartum period. In this study, changes of rumination time, reticuloruminal pH and temperature were investigated in eutocic (EUT, n = 10) and dystocic (DYS, n = 8) dairy cows around parturition. Rumination time was continuously recorded using an acoustic biotelemetry system, whereas reticuloruminal pH and temperature were recorded using an indwelling and wireless data transmitting system. The recording period lasted from 3 d before calving until 7 days in milk. For the comparison of rumination time and reticuloruminal characteristics between groups, time to return to baseline (the time interval required to return to baseline from the delivery of the calf) and area under the curve (AUC, both for prepartum and postpartum periods) were calculated for each parameter. Rumination time decreased from baseline 28 h before calving both for EUT and DYS cows (P = 0.023 and P = 0.017, respectively). After 20 h before calving, it decreased onwards to reach 32.4 ± 2.3 and 13.2 ± 2.0 min/4 h between 8 and 4 h before delivery in EUT and DYS cows, respectively, and then it decreased below 10 and 5 min during the last 4 h before calving (P = 0.003 and P = 0.008, respectively). Until 12 h after delivery rumination time reached 42.6 ± 2.7 and 51.0 ± 3.1 min/4 h in DYS and EUT dams, respectively, however, AUC and time to return to baseline suggested lower rumination activity in DYS cows than in EUT dams for the 168-h postpartum observational period (P = 0.012 and P = 0.002, respectively). Reticuloruminal pH decreased from baseline 56 h before calving both for EUT and DYS cows (P = 0.012 and P = 0.016, respectively), but did not differ between groups before delivery. In DYS cows, reticuloruminal temperature decreased from baseline 32 h before calving by 0.23 ± 0.02 °C (P = 0.012), whereas in EUT cows such a decrease was found only 20 h before delivery (0.48 ± 0.05 °C, P < 0.01). AUC of reticuloruminal temperature calculated for the prepartum period was greater in EUT cows than in DYS cows (P = 0.042). During the first 4 h after calving, it decreased from 39.7 ± 0.1 to 39.00 ± 0.1 °C and from 39.8 ± 0.1 to 38.8 ± 0.1 °C in EUT and DYS cows, respectively (P < 0.01 for both groups) and reached baseline levels after 35.4 ± 3.4 and 37.8 ± 4.2 h after calving in EUT and DYS cows, respectively. Based on our results, continuous monitoring of changes in rumination time and reticuloruminal temperature seems to be promising in the early detection of cows with a higher risk of dystocia. Depressed postpartum rumination time of DYS cows highlights the importance of the monitoring of cows experiencing difficulties at calving.

Keywords: reticuloruminal pH, reticuloruminal temperature, rumination time, dairy cows, dystocia

Procedia PDF Downloads 301
532 Functional Cell Surface Display Using Ice Nucleation Protein from Erwina ananas on Escherischia coli

Authors: Mei Yuin Joanne Wee, Rosli Md. Illias

Abstract:

Cell surface display is the expression of a protein with an anchoring motif on the surface of the cell. This approach offers advantages when used in bioconversion in terms of easier purification steps and more efficient enzymatic reaction. A surface display system using ice nucleation protein (InaA) from Erwina ananas as an anchoring motif has been constructed to display xylanase (xyl) on the surface of Escherischia coli. The InaA was truncated so that it is made up of the N- and C-terminal domain (INPANC-xyl) and it has successfully directed xylanase to the surface of the cell. A study was also done on xylanase fused to two other ice nucleation proteins, InaK (INPKNC-xyl) and InaZ (INPZNC-xyl) from Pseudomonas syringae KCTC 1832 and Pseudomonas syringae S203 respectively. Surface localization of the fusion protein was verified using SDS-PAGE and Western blot on the cell fractions and all anchoring motifs were successfully displayed on the outer membrane of E. coli. Upon comparison, whole-cell activity of INPANC-xyl was more than six and five times higher than INPKNC-xyl and INPZNC-xyl respectively. Furthermore, the expression of INPANC-xyl on the surface of E. coli did not inhibit the growth of the cell. This is the first report of surface display system using ice nucleation protein, InaA from E. ananas. From this study, this anchoring motif offers an attractive alternative to the current surface display systems.

Keywords: cell surface display, Escherischia coli, ice nucleation protein, xylanase

Procedia PDF Downloads 372
531 Cytotoxicity of Thymoquinone Alone or in Combination with Cisplatin (CDDP) Against Oral Squamous Cell Carcinoma in Vitro

Authors: Omar M. Al Aufi, Abdulwahab Noorwali, Ahmed Al Abd, Safia Alattas, Fathya Zahran, Fahd Almutairi

Abstract:

Cisplatin (CDDP) is a potent anticancer agent used for several tumor types. Thymoquinone (TQ) is a naturally occurring compound drawing great attention as an anticancer and chemomodulator for chemotherapies. Herein, we studied the potential cytotoxicity of thymoquinone, CDDP and their combination against human oral squamous cell carcinoma cells in contrast to normal oral epithelial cells. CDDP similarly killed both head and neck squamous cell carcinoma cells (UMSCC-14C) and normal oral epithelial cells (OEC). TQ alone exerted considerable cytotoxicity against UMSCC-14C cells, while it induced a weaker killing effect against normal oral epithelial cells (OEC). The equitoxic combination of TQ and CDDP showed additive to synergistic interaction against both UMSCC-14C and OEC cells. TQ alone increased apoptotic cell fraction in UMSCC-14C cells as early as after 6 hours. In addition, prolonged exposure of UMSCC-14C to TQ alone resulted in 96.7±1.6% total apoptosis, which was increased after combination with CDDP to 99.3±1.2% in UMSCC-14C cells. On the other hand, TQ induced a marginal increase in the apoptosis in OEC and even decreased the apoptosis induced by CDDP alone. Finally, apoptosis induction results were confirmed by the change in the expression levels of p53, Bcl-2 and Caspase-9 proteins in both UMSCC-14c and OEC cells.

Keywords: thymoquinone, cisplatin, apoptosis, oral squamous cell carcinoma, P53, Caspase-9, Bcl-2

Procedia PDF Downloads 41
530 Breastfeeding Knowledge, Attitudes and Practice: A Cross-Sectional Study among a Sample of Tunisian Mothers

Authors: Arfaoui Emna, Nouira Mariem

Abstract:

Background and aims: Breastfeeding is the reference feeding for a child, especially during the first months of life. It is not widespread in many countries due to many factors. There has been a decline in exclusive breastfeeding (EB) practice, particularly in the middle- and low-income countries, i.e., Tunisia. The aim of our study was to describe the knowledge, attitudes, and practice of a sample of Tunisian mothers toward breastfeeding. Methods: It was a descriptive cross-sectional study conducted during the year 2022 over a period of two months in three health structures in the north of Tunisia among mothers of infants aged 2 to 18 months. Levels of mothers’ knowledge (low/moderate/high) were determined using a score ranging from 0 to 11 points. EB was defined as the proportion of infants who were exclusively breastfed during the first six months of life. Results: A total of 180 women with a mean age of 33±4.9 years were included. The average knowledge score was equal to 6.4 ±1.5 points, with extremes ranging from 3 to 11 points. Most of the respondents had a moderate knowledge level (44.4%). More than half of surveyed mothers (66.1%) thought that breastfeeding deforms breasts, and 16.7% thought that breastfeeding is specific to women who do not work. Breastfeeding experience during the first week of life was considered difficult in 70% of cases. The prevalence of EB up to 6 months of age was equal to 16.4% [10.8-23.2]. The main reported obstacles during breastfeeding practice were having an insufficient quantity of breast milk (18.3%) and child difficulties with sucking (12.8%), and having pain in the breast while breastfeeding (12.80%). Conclusion: Our results highlighted the insufficient level of knowledge and a low prevalence of EB in our study population. Improving mothers’ knowledge and promoting EB practice is needed. Implementing health education strategies involving healthcare workers, who represent a main actor in education and breastfeeding promotion, is very important to reach a satisfactory frequency for EB.

Keywords: breastfeeding, practices, knowledge, Tunisia

Procedia PDF Downloads 61
529 The Effect of Porous Alkali Activated Material Composition on Buffer Capacity in Bioreactors

Authors: Girts Bumanis, Diana Bajare

Abstract:

With demand for primary energy continuously growing, search for renewable and efficient energy sources has been high on agenda of our society. One of the most promising energy sources is biogas technology. Residues coming from dairy industry and milk processing could be used in biogas production; however, low efficiency and high cost impede wide application of such technology. One of the main problems is management and conversion of organic residues through the anaerobic digestion process which is characterized by acidic environment due to the low whey pH (<6) whereas additional pH control system is required. Low buffering capacity of whey is responsible for the rapid acidification in biological treatments; therefore alkali activated material is a promising solution of this problem. Alkali activated material is formed using SiO2 and Al2O3 rich materials under highly alkaline solution. After material structure forming process is completed, free alkalis remain in the structure of materials which are available for leaching and could provide buffer capacity potential. In this research porous alkali activated material was investigated. Highly porous material structure ensures gradual leaching of alkalis during time which is important in biogas digestion process. Research of mixture composition and SiO2/Na2O and SiO2/Al2O ratio was studied to test the buffer capacity potential of alkali activated material. This research has proved that by changing molar ratio of components it is possible to obtain a material with different buffer capacity, and this novel material was seen to have considerable potential for using it in processes where buffer capacity and pH control is vitally important.

Keywords: alkaline material, buffer capacity, biogas production, bioreactors

Procedia PDF Downloads 234
528 Effect of Heat Stress on the Physiology of the Cork Oak

Authors: J. Zekri, N. Souilah, W. Abdelaziz, D. Alatou

Abstract:

Our study shall focus on the ability of trees cork oak that showed vis-à-vis sensitivity to climate change, including late spring frosts. The combination of these factors resulted in damage alarmed, therefore forest ecosystems weakened trees that can affect their ability to support other abiotic and biotic stresses, For this we tested its tolerance to thermal variations and cold weather conditions by estimating some stress markers (quantification of proteins, RNA, soluble sugars) that are quantified to evaluate the cold tolerance of seedlings. Sowing of cork oak (Quercus suber L.) is grown in controlled conditions at 25° C ± 2° C in long days 16h. These seedlings are transferred at low temperatures between 5° C and -6° C for a period of 3 hours. Biochemical analyzes were performed in the various organs of the cork oak seedlings. Cool temperatures induced a significant accumulation of proline in different organs of seedlings and the optimum concentrations were observed in the roots with very high concentrations (4 times larger than those of the control). The accumulation of soluble sugars is significantly in stems and roots at 0° C. Protein concentrations are very high in leaves of both growth and high waves in rod at -4° C to -2° C. Tolerance cork oak seems to be at the thermal limit of -2°C. The concentration of these metabolites in the various organs showed the ability oak cork hardening during the winter.

Keywords: climate change, thermal change, semi-aride, biochemical markers, heat stress

Procedia PDF Downloads 226
527 Characteristics and Quality of Chilean Abalone Undergoing Different Drying Emerging Technologies

Authors: Mario Pérez-Won, Anais Palma-Acevedo, Luis González-Cavieres, Roberto Lemus-Mondaca, Gipsy Tabilo-Munizaga

Abstract:

The Chilean abalone (Concholepas Concholepas) is a gastropod mollusk; it has a high commercial value due to the qualities of its meat, especially hardness, as a critical acceptance parameter. However, its main problem is its short shelf-life which is usually extended using traditional technologies with high energy consumption. Therefore, applying different technologies for the pre-treatment and drying process is necessary. In this research, pulsed electric field (PEF) was used as a pre-treatment for vacuum microwave drying (VMD), freeze-drying (FD), and hot-air drying (HAD). Drying conditions and characteristics were set according to previous experiments. The Drying samples were analyzed in terms of physical quality (color, texture, microstructure, and rehydration capacity), protein quality (degree of hydrolysis and computer protein efficiency ratio), and energy parameters. Regarding quality, the treatment that obtained lower harness was PEF+FD (195 N ± 10), the lowest change of color was for treatment PEF+VMD (ΔE: 17 ± 1.5), and the best rehydration capacity was for treatment PEF+VMD (1.2 h for equilibrium). For protein quality, the highest Computer-Protein Efficiency Ratio was the sample 2.0 kV/ cm of PEF (index of 4.18 ± 0.26 at the end of the digestion). Moreover, about energetic consumption, results show that VMD decreases the drying process by 97% whether PEF was used or not. Consequently, it is possible to conclude that using PEF as a pre-treatment for VMD and FD treatments has advantages that must be used following the consumer’s needs or preferences.

Keywords: chilean abalone, freeze-drying, proteins, pulsed electric fields

Procedia PDF Downloads 97
526 Probiotic Antibacterial Test of Pediococcus pentosaceus Isolated from Dadih in Inhibiting Periodontitis Bacteria: In Vitro Study on Bacteria Aggregatibacter actinomycetemcomitans

Authors: Nurlaili Syafar Wulan, Almurdi, Suprianto Kosno

Abstract:

Introduction: Periodontitis defined as an inflammatory disease of teeth supporting tissue with irritation of specific pathogens as the main aetiology. Periodontitis can be cured by giving medical action accompanied by administration of an antibiotic, but the use of antibiotic has a side effect that can cause bacterial resistance. This side effect can be corrected by probiotic, which has antibiotic-like substance but do not have bacterial resistance effect; it makes probiotic became a promising future periodontitis medication. West Sumatran people has their own typical traditional food product made from fermented buffalo’s milk called dadih, and it contained probiotics. Objectives: The aim of this study was to determine the ability of probiotic Pediococcus pentosaceus isolated from dadih in inhibiting the growth of bacteria Aggregatibacter actinomycetemcomitans. Material and Method: This was a true experimental study with post-test and control group design. This study was conducted on 36 samples of 2 treatment groups, the test group with probiotic Pediococcus pentosacesus isolated from dadih and the negative control group with sterile aquadest. The antibacterial effect was tested using the Kirby-Bauer disk diffusion method and calculated by measuring the zone of inhibition on MHA around paper disk using a sliding caliper with 0.5 mm accuracy. Result: The result of bivariate analysis using Independent t-test was p=0.00 where p < 0.05 means that there is a significant difference between the tested group and negative control group. Conclusion: Probiotic Pediococcus pentosaceus isolated from dadih are able to inhibit the growth of Aggregatibacter actinomycetemcomitans.

Keywords: aggregatibacter actinomycetemcomitans, antibacterial activities, periodontitis, probiotic Pediococcus pentosaceus

Procedia PDF Downloads 118
525 Novel Molecular Mechanisms Involved in Macrophage Phenotypic Polarization

Authors: Mansi Srivastava, Uzma Saqib, Adnan Naim, Anjali Roy, Dongfang Liu, Deepak Bhatnagar, Ravinder Ravinder, Mirza S. Baig

Abstract:

Macrophages polarize to proinflammatory M1 or anti-inflammatory M2 states with distinct physiological functions. This transition within the M1 to M2 phenotypes decides the nature, duration, and severity of an inflammatory response. However, inspite of a substantial understanding of the fate of these phenotypes, the underlying molecular mechanisms are not well understood. We have investigated the role of Neuronal nitric oxide synthase (NOS1) mediated regulation of Activator protein 1 (AP-1) transcription factor in macrophages as a critical effector of macrophage phenotypic change. Activator protein 1 (AP-1) is a group of dimeric transcription factors composed of jun, Fos, and ATF family proteins. We determined that NOS1-derived nitric oxide (NO) facilitate Fos and jun interaction which induces IL12 & IL23 expression. Pharmacological inhibition of NOS1 inhibits Fos and jun interaction but increases ATF2 and Fos dimerization. Switching of Fos and jun dimer to ATF2 and jun dimerization switches phenotype from IL–12high IL-23high IL-10low to IL–12low IL-23lowIL-10high phenotype, respectively. Together, these findings highlight a key role of the TLR4-NOS1-AP1 signaling axis in regulating macrophage polarization.

Keywords: inflammation, macrophage, lipopolysaccharide (LPS), proinflammatory cytokines, activator protein 1 (AP-1), neuronal nitric oxide synthase (NOS1)

Procedia PDF Downloads 268
524 Protein Quality of Game Meat Hunted in Latvia

Authors: Vita Strazdina, Aleksandrs Jemeljanovs, Vita Sterna

Abstract:

Not all proteins have the same nutritional value, since protein quality strongly depends on its amino acid composition and digestibility. The meat of game animals could be a high protein source because of its well-balanced essential amino acids composition. Investigations about biochemical composition of game meat such as wild boar (Sus scrofa scrofa), roe deer (Capreolus capreolus) and beaver (Castor fiber) are not very much. Therefore, the aim of the investigation was evaluate protein composition of game meat hunted in Latvia. The biochemical analysis, evaluation of connective tissue and essential amino acids in meat samples were done, the amino acids score were calculate. Results of analysis showed that protein content 20.88-22.05% of all types of meat samples is not different statistically. The content of connective tissue from 1.3% in roe deer till 1.5% in beaver meat allowed classified game animal as high quality meat. The sum of essential amino acids in game meat samples were determined 7.05–8.26g100g-1. Roe deer meat has highest protein content and lowest content of connective tissues among game meat hunted in Latvia. Concluded that amino acid score for limiting amino acids phenylalanine and tyrosine is high and shows high biological value of game meat.

Keywords: dietic product, game meat, amino acids, scores

Procedia PDF Downloads 300
523 The Immunosuppressive Effects of Silymarin with Rapamaycin on the Proliferation and Apoptosis of T Cell

Authors: Nahid Eskandari, Marjan Ghagozolo, Ehsan Almasi

Abstract:

Introduction: Silymarin, as a polyphenolic flavonoid derived from milk thistle (Silybum marianum), is known to have antioxidant, immunomodulatory, antiproliferative, antifibrotic, and antiviral effects. The goal of this study was to determine immunosuppressive effect of Silymarin on proliferation and apoptosis of human T cells in comparison with Rapamycin and FK506. Methods: Peripheral Blood Mononuclear Cells (PBMCs) from healthy individuals were activated with Con A (5µg/ml) and then treated with Silymarin, Rapamycin and FK506 in various concentrations (0.001, 0.01, 0.1, 1, 10,100 and 200M) for 5 days. PBMCs were examined for proliferation using CFSE assay and the concentration that inhibited 50% of the cell proliferation (IC50) was determined for each treatment. For apoptosis assay using flow cytometry, PBMCs were activated with Con A and treated with IC50 dose of Silymarin, Rapamycin and FK506 for 5 days, then cell apoptosis was analysed by FITC-annexin V/PI staining and flow cytometry. The effects of Silymarin, Rapamycin and FK506 on the activation of PARP (poly ADP ribose polymerase) pathway in PBMCs stimulated with Con A and treated with IC50 dose of drugs for 5 days evaluated using the PathScan cleaved PARP sandwich ELISA kit. Results: This study showed that Silymarin had the ability to inhibit T cell proliferation in vitro. Moreover, our results indicated that 100 μM (P < 0.001) and 200 μM (P < 0.001) of Silymarin has more inhibitory effect on T cells proliferation than FK506 and Rapamycin. Our data showed that the effective doses (IC50) of Silymarin, FK506 and Rapamycin were 3×10-5 µM, 10-8 µM and 10-6 µM respectively. Data showed that the inhibitory effect of Silymarin, FK506 and Rapamycin on T cell proliferation was not due to cytotoxicity and none of these drugs at IC50 concentration had not affected the level of cleaved PARP. Conclusion: Silymarin could be a good candidate for immunosuppressive therapy for certain medical conditions with superior efficacy and lesser toxicity in comparison with other immunosuppressive drugs.

Keywords: silymarin, immunosuppressive effect, rapamycin, immunology

Procedia PDF Downloads 248
522 Multiple Strategies in Prevention of Metabolic Syndrome Result from Vitamin D Deficiency in Children

Authors: Maryam Ghavam Sadri, Maryam Shahrooz

Abstract:

Background: Nowadays the prevalence of metabolic syndrome (Mets) has taken on a growing trend. Studies have shown the relationship between vitamin D deficiency (VDD) status and Mets in children. Also studies have recorded that exerting strategies for vitamin D status improvement can help prevent Mets in children. This study investigated multiple strategies of prevention of Mets resulting from VDD in children. Methods: This review study has been done by using keywords related to the topic and 54 articles were found (2000-2015) that 25 were selected according to the indicators of Mets, supplementation and fortification of foods with vitamin D and attention to children environment and life style. Results: Studies have suggested the correlation between serum levels of vitamin D with waist circumference (p < 0.0001), systolic blood pressure (p=0.01), HOMA-IR (p=0.001) and HDL cholesterol (p < 0.0001). An inverse correlation between serum 25 (OH) D and HOMA-IR (p = 0.006) and insulin (P = 0.002) has been proved in overweight group. Higher HOMASDS and triglycerides found in vitamin D deficient obese children compared to control group without VDD (p=0.04). After supplementation with vitamin D, serum TG concentration decreases significantly (p=0.04), and improves insulin resistance (p=0.02). The prevalence of VDD is associated with time of watching TV (P < 0.01), hours of physical activity per week (P = 0.01), skipping breakfast (P < 0.001) soda intake (P < 0.001), and milk intake per day (P < 0.01). Conclusion: According to the beneficial role of vitamin D in prevention of Mets and proven relationship between serum levels of vitamin D and Mets indicators, we can prevent childhood Mets through the application of appropriate strategies such as supplementation and food fortification with vitamin D and positive changes in children life style with especial attention to physical activity in exposure of sunlight and their environment condition.

Keywords: children, metabolic syndrome, prevention strategies, vitamin D

Procedia PDF Downloads 550
521 Multiplying Vulnerability of Child Health Outcome and Food Diversity in India

Authors: Mukesh Ravi Raushan

Abstract:

Despite consideration of obesity as a deadly public health issue contributing 2.6 million deaths worldwide every year developing country like India is facing malnutrition and it is more common than in Sub-Saharan Africa. About one in every three malnourished children in the world lives in India. The paper assess the nutritional health among children using data from total number of 43737 infant and young children aged 0-59 months (µ = 29.54; SD = 17.21) of the selected households by National Family Health Survey, 2005-06. The wasting was measured by a Z-score of standardized weight-for-height according to the WHO child growth standards. The impact of education with place of residence was found to be significantly associated with the complementary food diversity score (CFDS) in India. The education of mother was positively associated with the CFDS but the degree of performance was lower in rural India than their counterpart from urban. The result of binary logistic regression on wasting with WHO seven types of recommended food for children in India suggest that child who consumed the milk product food (OR: 0.87, p<0.0001) were less likely to be malnourished than their counterparts who did not consume, whereas, in case of other food items as the child who consumed food product of seed (OR: 0.75, p<0.0001) were less likely to be malnourished than those who did not. The nutritional status among children were negatively associated with the protein containing complementary food given the child as those child who received pulse in last 24 hour were less likely to be wasted (OR: 0.87, p<0.00001) as compared to the reference categories. The frequency to feed the indexed child increases by 10 per cent the expected change in child health outcome in terms of wasting decreases by 2 per cent in India when place of residence, education, religion, and birth order were controlled. The index gets improved as the risk for malnutrition among children in India decreases.

Keywords: CFDS, food diversity index, India, logistic regression

Procedia PDF Downloads 238
520 Comparative Rumen Degradable and Rumen Undegradable Fractions in Untreated, Formaldehyde and Heat Treated Vegetable Protein Sources of Pakistan

Authors: Illahi Bakhsh Marghazani, Nasrullah, Masood Ul Haq Kakar, Abdul Hameed Baloch, Ahmad Nawaz Khoso, Behram Chacher

Abstract:

Protein sources are the major part of ration fed to dairy buffaloes in Pakistan however, the limited availability and lack of judicious use of protein resources are further aggravating the conditions to enhance milk and meat production. In order to gain maximum production from limited protein source availability, it is necessary to balance feed for rumen degradable and rumen undegradable protein fractions. This study planned to know the rumen degradable and rumen undegradable fractions in all vegetable protein sources with (formaldehyde and heat treatment) and without treatments. Samples of soybean meal, corn gluten meal 60%, maize gluten feed, guar meal, sunflower meal, rapeseed meal, rapeseed cake, canola meal, cottonseed cake, cottonseed meal, coconut cake, coconut meal, palm kernel cake, almond cake and sesame cake were collected from ten different geographical locations of Pakistan. These samples were also subjected to formaldehyde (1% /100g CP of test feed) and heat treatments (1 hr at 15 lb psi/100 g CP of test feed). In situ technique was used to know the ruminal degradability characteristics. Data obtained were fitted to Orskove equation. Results showed that both treatments significantly (P < 0.05) decreased ruminal degradability in all vegetable protein sources than untreated vegetable protein sources, however, of both treatments, heat treatment was more effective than formaldehyde treatment in decreasing ruminal degradability in most of the studied vegetable protein sources.

Keywords: formaldehyde and heat treatments, in situ technique, rumen degradable and rumen undegradable fractions, vegetable protein sources

Procedia PDF Downloads 313
519 Apoptosis Activity of Persea declinata (Bl.) Kosterm Bark Methanolic Crude Extract

Authors: P. Narrima, C. Y. Looi, M. A. Mohd, H. M. Ali

Abstract:

Persea declinata (Bl.) Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill), which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl.) Kosterm bark methanolic crude extract (PDM). PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 µg/mL after 48h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH) release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development.

Keywords: antiproliferative, apoptosis, MCF-7 human breast cancer, Persea declinata

Procedia PDF Downloads 232
518 Simultaneous Targeting of MYD88 and Nur77 as an Effective Approach for the Treatment of Inflammatory Diseases

Authors: Uzma Saqib, Mirza S. Baig

Abstract:

Myeloid differentiation primary response protein 88 (MYD88) has long been considered a central player in the inflammatory pathway. Recent studies clearly suggest that it is an important therapeutic target in inflammation. On the other hand, a recent study on the interaction between the orphan nuclear receptor (Nur77) and p38α, leading to increased lipopolysaccharide-induced hyperinflammatory response, suggests this binary complex as a therapeutic target. In this study, we have designed inhibitors that can inhibit both MYD88 and Nur77 at the same time. Since both MYD88 and Nur77 are an integral part of the pathways involving lipopolysaccharide-induced activation of NF-κB-mediated inflammation, we tried to target both proteins with the same library in order to retrieve compounds having dual inhibitory properties. To perform this, we developed a homodimeric model of MYD88 and, along with the crystal structure of Nur77, screened a virtual library of compounds from the traditional Chinese medicine database containing ~61,000 compounds. We analyzed the resulting hits for their efficacy for dual binding and probed them for developing a common pharmacophore model that could be used as a prototype to screen compound libraries as well as to guide combinatorial library design to search for ideal dual-target inhibitors. Thus, our study explores the identification of novel leads having dual inhibiting effects due to binding to both MYD88 and Nur77 targets.

Keywords: drug design, Nur77, MYD88, inflammation

Procedia PDF Downloads 287
517 Clustered Regularly Interspaced Short Palindromic Repeat/cas9-Based Lateral Flow and Fluorescence Diagnostics for Rapid Pathogen Detection

Authors: Mark Osborn

Abstract:

Clustered, regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. Commercially available reagents were integrated into a CRISPR/Cas9-based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. A rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction was also developed. These findings provide proof of principle for CRISPR/Cas9 point-of-care diagnosis that can detect specific SARS-CoV-2 strain(s). Further, Cas9 cleavage allows for a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology. Collectively, this approach is a facile platform for diagnostics with broad application to user-defined sequence interrogation and detection.

Keywords: CRISPR/Cas9, lateral flow assay, SARS-Co-V2, single-nucleotide resolution

Procedia PDF Downloads 167
516 Development of Enzymatic Amperometric Biosensors with Carbon Nanotubes Decorated with Iron Oxide Nanoparticles

Authors: Uc-Cayetano E. G., Ake-Uh O. E., Villanueva-Mena I. E., Ordonez L. C.

Abstract:

Carbon nanotubes (CNTs) and other graphitic nanostructures are materials with extraordinary physical, physicochemical and electrochemical properties which are being aggressively investigated for a variety of sensing applications. Thus, sensing of biological molecules such as proteins, DNA, glucose and other enzymes using either single wall or multiwall carbon nanotubes (MWCNTs) has been widely reported. Despite the current progress in this area, the electrochemical response of CNTs used in a variety of sensing arrangements still needs to be improved. An alternative towards the enhancement of this CNTs' electrochemical response is to chemically (or physically) modify its surface. The influence of the decoration with iron oxide nanoparticles in different types of MWCNTs on the amperometric sensing of glucose, urea, and cholesterol in solution is investigated. Commercial MWCNTs were oxidized in acid media and subsequently decorated with iron oxide nanoparticles; finally, the enzymes glucose oxidase, urease, and cholesterol oxidase are chemically immobilized to oxidized and decorated MWCNTs for glucose, urease, and cholesterol electrochemical sensing. The results of the electrochemical characterizations consistently show that the presence of iron oxide nanoparticles decorating the surface of MWCNTs enhance the amperometric response and the sensitivity to increments in glucose, urease, and cholesterol concentration when compared to non-decorated MWCNTs.

Keywords: WCNTs, enzymes, oxidation, decoration

Procedia PDF Downloads 110
515 Design of a Recombinant Expression System for Bacterial Cellulose Production

Authors: Gizem Buldum, Alexander Bismarck, Athanasios Mantalaris

Abstract:

Cellulose is the most abundant biopolymer on earth and it is currently being utilised in a multitude of industrial applications. Over the last 30 years, attention has been paid to the bacterial cellulose (BC), since BC exhibits unique physical, chemical and mechanical properties when compared to plant-based cellulose, including high purity and biocompatibility. Although Acetobacter xylinum is the most efficient producer of BC, it’s long doubling time results in insufficient yields of the cellulose production. This limits widespread and continued use of BC. In this study, E. coli BL21 (DE3) or E. coli HMS cells are selected as host organisms for the expression of bacterial cellulose synthase operon (bcs) of A.xylinum. The expression system is created based on pET-Duet1 and pCDF plasmid vectors, which carry bcs operon. The results showed that all bcs genes were successfully transferred and expressed in E.coli strains. The expressions of bcs proteins were shown by SDS and Native page analyses. The functionality of the bcs operon was proved by congo red binding assay. The effect of culturing temperature and the inducer concentration (IPTG) on cell growth and plasmid stability were monitored. The percentage of plasmid harboring cells induced with 0.025 mM IPTG was obtained as 85% at 22˚C in the end of 10-hr culturing period. It was confirmed that the high output cellulose production machinery of A.xylinum can be transferred into other organisms.

Keywords: bacterial cellulose, biopolymer, recombinant expression system, production

Procedia PDF Downloads 378
514 Genome Sequencing of the Yeast Saccharomyces cerevisiae Strain 202-3

Authors: Yina A. Cifuentes Triana, Andrés M. Pinzón Velásco, Marío E. Velásquez Lozano

Abstract:

In this work the sequencing and genome characterization of a natural isolate of Saccharomyces cerevisiae yeast (strain 202-3), identified with potential for the production of second generation ethanol from sugarcane bagasse hydrolysates is presented. This strain was selected because its capability to consume xylose during the fermentation of sugarcane bagasse hydrolysates, taking into account that many strains of S. cerevisiae are incapable of processing this sugar. This advantage and other prominent positive aspects during fermentation profiles evaluated in bagasse hydrolysates made the strain 202-3 a candidate strain to improve the production of second-generation ethanol, which was proposed as a first step to study the strain at the genomic level. The molecular characterization was carried out by genome sequencing with the Illumina HiSeq 2000 platform paired end; the assembly was performed with different programs, finally choosing the assembler ABYSS with kmer 89. Gene prediction was developed with the approach of hidden Markov models with Augustus. The genes identified were scored based on similarity with public databases of nucleotide and protein. Records were organized from ontological functions at different hierarchical levels, which identified central metabolic functions and roles of the S. cerevisiae strain 202-3, highlighting the presence of four possible new proteins, two of them probably associated with the positive consumption of xylose.

Keywords: cellulosic ethanol, Saccharomyces cerevisiae, genome sequencing, xylose consumption

Procedia PDF Downloads 306