Search results for: chick bean protein isolate

Authors: Adebisi M. Tiamiyu, Adewale F. Adeyemi, Olu-Ayobamikale V. Irewunmi

Abstract:

Fish consumption has increased in recent years in both developing and advanced countries, owing to increased awareness of its nutritional and therapeutic benefits and its availability and affordability relative to other animal protein sources. Fish and fish products, however, are extremely prone to contamination by a wide range of hazardous organic and inorganic substances. This study assessed the levels of three heavy metals, copper (Cu), iron (Fe), and zinc (Zn), in frozen fish imported into Nigeria and sold in Ondo City for their safety for human consumption as recommended by WHO and FEPA. Three species of frozen fish (Scombrus scombrus, Merluccius merluccius, and Clupea harengus) were purchased, and the wet tissues (gills, muscles, and liver) were digested using a 3:1 mixture of nitric acid (HNO3) and hydrochloric acid (HCL). An atomic absorption spectrophotometer (AAS) was used to detect the amount of metal in the tissues. The levels of heavy metals in different fish species' organs varied. The fish had Zn > Fe > Cu heavy metal concentrations in that order. While the concentration of Cu and Fe in the tissues of all three fish species studied were within the WHO and FEPA prescribed limits for food fish, the concentration of Zn in the muscles of M. merluccius (0.262±0.052), C. harengus harengus (0.327±0.099), and S. scombrus (0.362±0.119) was above the prescribed limit (0.075 ppm) set by FEPA. An excessive amount of zinc in the body can cause nausea, headaches, decreased immunity, and appetite loss.

Keywords: heavy metal, atomic absorption spectrophotometer, fish, agencies

Procedia PDF Downloads 72

Authors: Felicia Segeth, Florian G. Klein, Lea Berger, Andreas Kolk, Per S. Holm

Abstract:

The entry of oncolytic viruses (OVs) into clinical application opens groundbreaking changes in current and future treatment regimens. However, despite their potent anti-cancer activity in vitro, clinical studies revealed limitations of OVs as monotherapy. The same applies to CDK 4/6 inhibitors (CDK4/6i) targeting cell cycle as well as bromodomain and extra-terminal domain inhibitors (BETi) targeting gene expression. In this study, the anti-tumoral effect of XVir-N-31, an YB-1 dependent oncolytic adenovirus, was evaluated in combination with Ribociclib, a CDK4/6i, and JQ1, a BETi. The head and neck squamous cell carcinoma (HNSCC) cell lines Fadu, SAS, and Cal-33 were used. DNA replication and gene expression of XVir-N-31 was measured by RT-qPCR, protein expression by western blotting, and cell lysis by SRB assays. Treatment with CDK4/6i and BETi increased viral gene expression, viral DNA replication, and viral particle formation. The data show that the combination of oncolytic adenovirus XVir-N-31 with CDK4/6i & BETi acts highly synergistic in cancer cell lysis. Furthermore, additional molecular analyses on this subject demonstrate that the positive transcription elongation factor P-TEFb plays a decisive role in this regard, indicating an influence of the combinational therapy on gene transcription control. The combination of CDK4/6i & BETi and XVir-N-31 is an attractive strategy to achieve substantial cancer cell killing and is highly suitable for clinical testing.

Keywords: adenovirus, BET, CDK4/6, HNSCC, P-TEFb, YB-1

Procedia PDF Downloads 119

Authors: Elad Arad, Raz Jelinek, Hanna Rapaport

Abstract:

Amyloidoses are a family of diseases characterized by abnormal protein folding that leads to aggregation. They accumulate to form fibrillar plaques which are implicated in the pathogenesis of Alzheimer, prion, diabetes type II and other diseases. To the best of our knowledge, despite extensive research efforts devoted to plaque aggregates inhibition, there is yet no cure for this phenomenon. Titanium and its alloys are found in growing interest for biomedical applications. Variety of surface modifications enable porous, adhesive, bioactive coatings for its surface. Titanium oxides (titania) are also being developed for photothermal and photodynamic treatments. Inspired by this, we set to explore the effect of functionalized titania nanoparticles in combination with external stimuli, as potential photothermal ablating agents against amyloids. Titania nanoparticles were coated with bi-functional catechol derivatives (dihydroxy-phenylalanine propanoic acid, noted DPA) to gain targeting properties. In conjunction with UV-radiation, these nanoparticles may selectively destroy the vicinity of their target. Titania modified 5 nm nanoparticles coated with DPA were further conjugated to the amyloid-targeting Congo Red (CR). These Titania-DPA-CR nanoparticles were found to target mature amyloid fibril of both amyloid-β (Aβ 1-42 a.a). Moreover, irradiation of the peptides in presence of the modified nanoparticles decreased the aggregate content and oligomer fraction. This work provides insights into the use of modified titania nanoparticles for amyloid plaque targeting and photothermal destruction. It may shed light on future modifications and functionalization of titania nanoparticles for different applications.

Keywords: titanium dioxide, amyloids, photothermal treatment, catechol, Congo-red

Procedia PDF Downloads 147

Authors: Ben Abderrahmane Ayoub El Fateh, Bnina Rachid

Abstract:

A comprehensive comparison of antioxidant defenses in the dermis and epidermis and their response to exposure to ultraviolet (UV) irradiation has not previously been attempted. In this study, enzymic and non-enzymic antioxidants in epidermis and dermis of hairless mice were compared. Enzyme activities are presented both as units/gram of skin and units/milligram of protein; arguments are presented for the superiority of skin wet weight as a reference base. Catalase, glutathione peroxidase, and glutathione reductase (units/gram of skin) were higher in the epidermis than dermis by 49%, 86%, and 74%, respectively. Superoxide dismutase did not follow this pattern. Lipophilic antioxidants ( -tocopherol, ubiquinol 9, and ubiquinone 9) and hydrophilic antioxidants (ascorbic acid, dehydroascorbic acid, and glutathione) were 24–95% higher in the epidermis than in dermis. In contrast, oxidized glutathione was 60% lower in the epidermis than in dermis. Mice were irradiated with solar light to examine the response of these cutaneous layers to UV irradiation. After irradiation with 25 J/cm2 (UVA + UVB, from a solar simulator), 10 times the minimum erythemal dose, epidermal and dermal catalase and superoxide dismutase activities were greatly decreased. Tocopherol, ubiquinol 9, ubiquinone 9, ascorbic acid, dehydroascorbic acid, and reduced glutathione decreased in both epidermis and dermis by 26-93%. Oxidizedgiutathione showed a slight, non-significant increase. Because the reduction in total ascorbate and catalase was much more severe in the epidermis than dermis, it can be concluded that UV light is more damaging to the antioxidant defenses in the epidermis than in the dermis.

Keywords: antioxidant defenses, enzymic, epidermis, oxidizedgiutathione

Procedia PDF Downloads 441

Authors: Kolawole F. Farinloye, Samson O. Ojo

Abstract:

Two Columbiformes species were recently introduced to the newly established Awba Dam Tourism Centre [ADTC], hence there is need to investigate the effect of seasonal variation on these species with respect to hematological composition. Blood samples were obtained from superficial ulna vein of the 128 apparently healthy C. livia and C. guinea into tubes containing EDTA as anticoagulant. Thin blood smears (TBS) were prepared, stained and viewed under microscope. Values of Red Blood Cell (RBC) count, White Blood Cell (WBC) count, cholesterol (CH), Uric Acid (UA), Protein (PR), Mean Corpuscular Volume (MCV), Haemoglobin Content (HB), Blood Volume (BV), Plasma Glucose (PG) and Length/Width (L/W) ratio of red blood cells were assessed. The procedure was carried out on a seasonal basis (wet and dry seasons of 2013-2014). Data was analyzed using descriptive and inferential statistics. Lymphocyte count for C. livia was F3, 161 = 13.15, while for C. guinea was F3, 178 = 13.15. Heterophil, H/L ratio and Muscle score values for both species were (rs = -0.38, rs = -0.44), (rs = 0.51, rs = 0.31) (4, 3) respectively. Analyses also demonstrated a low WBC to RBC ratio (0.004: 25.3) in both species during the wet season compared to dry season, respectively. L/W varied significantly among sampling seasons i.e. wet (19.1% of BV, 12.6% of BV, 0.1% of BV) and dry (18.9% of BV, 12.7% of BV, 0.08% of BV). The level of HB in wet season (19.20±8.46108) is lower compared to dry season (19.70±8.48762). T-test also showed (wet=15.625, 0.111), (dry=12.125, 0.146) respectively, hence there is no association between species and haematological parameters. Species introduced were found to be haematologically stable. Although there were slight differences in seasonal composition, however this can be attributed to seasonal variation; suggesting little or no effect of seasons on their blood composition.

Keywords: seasonal variation, Columbiformes, Awba Dam tourism centre, University of Ibadan, Ibadan

Procedia PDF Downloads 343

Authors: Brian Marsh

Abstract:

Nitrogen fertilizer is the most used and often the most mismanaged nutrient input. Nitrogen management has tremendous implications on crop productivity, quality and environmental stewardship. Sufficient nitrogen is needed to optimum yield and quality. Soil and in-season plant tissue testing for nitrogen status are a time consuming and expensive process. Real time sensing of plant nitrogen status can be a useful tool in managing nitrogen inputs. The objectives of this project were to assess the reliability of remotely sensed non-destructive plant nitrogen measurements compared to wet chemistry data from sampled plant tissue, develop in-season nitrogen recommendations based on remotely sensed data for improved nitrogen use efficiency and assess the potential for determining yield and quality from remotely sensed data. Very good correlations were observed between early-season remotely sensed crop nitrogen status and plant nitrogen concentrations and subsequent in-season fertilizer recommendations. The transmittance/absorbance type meters gave the most accurate readings. Early in-season fertilizer recommendation would be to apply 40 kg nitrogen per hectare plus 16 kg nitrogen per hectare for each unit difference measured with the SPAD meter between the crop and reference area or 25 kg plus 13 kg per hectare for each unit difference measured with the CCM 200. Once the crop was sufficiently fertilized meter readings became inconclusive and were of no benefit for determining nitrogen status, silage yield and quality and grain yield and protein.

Keywords: wheat, nitrogen fertilization, chlorophyll meter

Procedia PDF Downloads 394

Authors: P. Serpytis, K. Azukaitis, U. Gargalskaite, R. Navickas, J. Badariene, V. Dzenkeviciute

Abstract:

Purpose: Total longitudinal displacement (tLoD) of the common carotid artery (CCA) wall is a novel ultrasound marker of vascular function that can be evaluated using modified speckle tracking techniques. Decreased CCA tLoD has already been shown to be associated with diabetes and was shown to predict one year cardiovascular outcome in patients with suspected coronary artery disease (CAD) . The aim of our study was to evaluate if CCA tLoD differ between patients with moderate or high cardiovascular (CV) risk and patients after recent acute myocardial infarction (AMI). Methods: 49 patients (54±6 years) with moderate or high CV risk and 42 patients (58±7 years) after recent AMI were included. All patients were non-diabetic. CCA tLoD was evaluated using GE EchoPAC speckle tracking software and expressed as mean of both sides. Data on systolic blood pressure, total and high density lipoprotein (HDL) cholesterol levels, high sensitivity C-reactive protein (hsCRP) level, smoking status and family history of early CV events was evaluated and assessed for association with CCA tLoD. Results: tLoD of CCA did not differ between patients with moderate or high CV risk and patients with very high CV risk after MI (0.265±0.128 mm vs. 0.237±0.103 mm, p>0.05). Lower tLoD was associated with lower HDL cholesterol levels (r=0.211, p=0.04) and male sex (0.228±0.1 vs. 0.297±0.134, p=0.01). Conclusions: tLoD of CCA did not differ between patients with moderate or high CV risk and patients with very high CV risk after AMI. However, lower CCA tLoD was significantly associated with low HDL cholesterol levels and male sex.

Keywords: total longitudinal displacement, carotid artery, cardiovascular risk, acute myocardial infarction

Procedia PDF Downloads 384

Authors: Jie Ding, Yingying Pan, Shammy Raj, Lindy Schaffrick, Jolene Wong, Antoinette Nguyen, Sharada Manchikanti, Larry Unsworth, Peter Kwan, Edward E. Tredget

Abstract:

Background: Exosomes (EXOs) have been considered a new target that is thought to be involved in and treat wound healing. More research is needed to fully understand the EXO characteristics and mechanisms of EXO-mediated wound healing, especially wound healing after burn injury. Methods: Total EXOs were isolated from 85 serum samples of 29 burn patients and 13 healthy individuals. We characterized the EXOs for morphology and density, serum concentration, protein level, marker expression, size distribution, and cytokine content. After confirmation of EXO uptake by dermal fibroblasts, we also explored functional regulation of primary human normal skin and hypertrophic scar fibroblast cell lines by the EXOs in vitro, including cell proliferation and apoptosis. Results: EXOs dynamically changed their morphology, density, size, and cytokine level during wound healing in burn patients, which were correlated with burn severity and the stages of wound healing. EXOs from both burn patients and healthy individuals stimulated dermal fibroblast proliferation and apoptosis. Conclusion: EXO features may be important signals that influence wound healing after burn injury; however, to understand the mechanisms by which EXOs regulated the fibroblasts in healing wounds, further studies will be required in the future.

Keywords: exosome, burn, wound healing, hypertrophic scarring, cytokines

Procedia PDF Downloads 83

Authors: Okolie Pius Ifeanyi, Emerenini Emilymary Chima

Abstract:

Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits using Molecular Nested PCR analysis and the efficacy of cell free supernatants from Lactic Acid Bacteria (LAB) isolated from fresh fruits for in vitro control of some tomato pathogens. Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples. The inhibitory potentials of supernatant obtained from LAB isolates of fruits origin that were molecularly characterized were investigated against some tomato phytopathogens using agar-well method with the view to develop biological agents for some tomato disease causing organisms. Methodology: Gram positive, catalase negative strains of LAB were isolated from fresh fruits on Man Rogosa and Sharpe agar (Lab M) using streaking method. Isolates obtained were molecularly characterized by means of genomic DNA extraction kit (Norgen Biotek, Canada) method. Standard methods were used for Nested Polymerase Chain Reaction (PCR) amplification targeting the 16S rRNA gene using universal 16S rRNA gene and LAB specific primers, agarose gel electrophoresis, purification and sequencing of generated Nested PCR products (Macrogen Inc., USA). The partial sequences obtained were identified by blasting in the non-redundant nucleotide database of National Center for Biotechnology Information (NCBI). The antimicrobial activities of characterized LAB against some tomato phytopathogenic bacteria which include (Xanthomonas campestries, Erwinia caratovora, and Pseudomonas syringae) were obtained by using the agar well diffusion method. Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%). The cell free supernatants of LAB from fresh fruits origin (Weissella cibaria, Weissella confusa, Leuconostoc paramensenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Lactobacillus pentosus) can inhibits these bacteria by creating clear zones of inhibition around the wells containing cell free supernatants of the above mentioned strains of lactic acid bacteria. Conclusion: This study shows that potentially LAB can be quickly characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer. Tomato disease causing organisms can be most likely biologically controlled by using extracts from LAB. This finding will reduce the potential hazard from the use of chemical herbicides on plant.

Keywords: nested pcr, molecular characterization, 16s rRNA gene, lactic acid bacteria

Procedia PDF Downloads 414

Authors: Muhammad Yousaf Ali, Shahid Mansoor, Javeria Qazi, Imran Amin, Musarrat Shaheen

Abstract:

Cotton leaf curl disease (CLCuD) is the major threat to the cotton crop and is transmitted by whitefly (Bemisia tabaci). Since multiple begomoviruses and associated satellites are involved in CLCuD, approaches based on the concept of broad-spectrum resistance are essential for effective disease control. Gro-El and G5 are two proteins from whitefly endosymbiont and M13 bacteriophage origin, respectively. Gro-El encapsulates the virus particle when it enters the whitefly and protects the virus from the immune system of the whitefly as well as prevents viral expression in it. This characteristic of Gro-El can be exploited to get resistance against viruses if expressed in plants. G5 is a single-stranded DNA binding protein, expression of which in transgenic plants will stop viral expression on its binding with ssDNA. The use of tissue-specific promoters is more efficient than constitutive promoters. Transgenics of Nicotiana benthamiana for Gro-El under constitutive promoter and Gro-El under phloem specific promoter were made. In comparison to non-transgenic plants, transgenic plants with Gro-El under NSP promoter showed promising results when challenged against cotton leaf curl Multan virus (CLCuMuV) along with cotton leaf curl Multan beta satellite (CLCuMB), cotton leaf curl Khokhran virus (CLCuKoV) along with cotton leaf curl Multan beta satellite (CLCuMB) and Pedilenthus leaf curl virus (PedLCV) along with Tobacco leaf curl beta satellite (TbLCB).

Keywords: cotton leaf curl disease, whitefly, endosymbionts, transgenic, resistance

Procedia PDF Downloads 98

Authors: Mohamed F. Ahmed, Mabruka S. Elashheb, Fatma M. Ben Rabha

Abstract:

This study aimed to determine the possible protective effects of L‐carnitine against gentamicin‐induced nephrotoxicity. Forty male albino rats were divided into 4 groups (10 rats each); Group 1: normal control, group 2: induced nephrotoxicity (gentamicin 50 mg/kg/day S.C; 8 days) , group 3: treated with L‐carnitine (40 mg/kg/d SC for 12 days) and group 4: treated with L‐carnitine 4 days before and for 8 days in concomitant with gentamicin. Gentamicin‐induced nephrotoxicity (group 2): caused significant increase in serum urea, creatinine, urinary N‐acetyl‐B‐D‐glucosaminidase (NAG), gamma glutamyl transpeptidase (GGT), urinary total protein and kidney tissue malondialdehyde (MDA) with significant decrease in serum superoxide dismutase (SOD), serum catalase and creatinine clearance and marked tubular necrosis in the proximal convoluted tubules with interruption in the basement membrane around the necrotic tubule compared to the normal control group. L‐carnitine 4 days before and for 8 days in concomitant with gentamicin (group 4) offered marked decrease in serum urea, serum creatinine, urinary NAG, urinary GGT, urinary proteins and kidney tissue MDA, with marked increase in serum SOD, serum catalase and creatinine clearance with marked improvement in the tubular damage compared to gentamicin‐induced nephrotoxicity group. L‐carnitine administered for 12 days produced no change in the above-mentioned parameters as compared to the normal control group. In conclusion: L‐carnitine could reduce most of the biochemical parameters and also improve the histopathological features of the kidney associated with gentamicin-induced nephrotoxicity.

Keywords: gentamicin, nephrotoxicity, L‐carnitine, kidney disease

Procedia PDF Downloads 359

Authors: Maryam Bordbar, Yaser Mokhayeri, Sajjad Roosta, Fatemeh Ghasemi, Saeed Choobkar, Hamidreza Nikbakht, Ebrahim Falahi

Abstract:

Objective: Diabetes mellitus type 2 is the most common metabolic disorder that is growing exponentially worldwide. Satureja Khuzestanica Jamzad is a native plant of Iran that grows widely in the south of Iran. Its antimicrobial, antioxidant, anti-inflammatory and pain-relieving effects have been documented in animal studies. The purpose of this study is to investigate the effect of consumption daily S. khuzestanica on inflammatory and antioxidant indicators in type 2 diabetic patients. Methods and Materials: In a double-blind, placebo-controlled clinical trial, 67 patients with type 2 diabetes were included and divided into two groups. One group received S. khuzestanica (capsule containing 500 mg) and the other group received placebo (500 mg talcum powder) once a day for 12 weeks. After the intervention, the inflammatory and antioxidant indicators of the two groups were compared. Results: In comparison to placebo groups, there was a significant difference in levels of total antioxidant capacity, superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase; these antioxidant indicators were higher in the intervention group (P<0.05). Moreover, a considerable decrease in weight, CRP and IL-6 levels were observed in patients in the S.Khuzestanica group. Conclusion: Our findings may provide novel complementary treatments without adverse effects for diabetes complications.

Keywords: Satureja khuzestanica Jamzad, diabetes mellitus, antioxidant indicators, IL-6, C-reactive protein

Procedia PDF Downloads 71

Authors: Mohd Sidek Ahmad, Zainon Mohd Noor, Zaidah Zainal Ariffin

Abstract:

Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Keywords: isolation, identification, fibrinolytic protease, endophytic fungi, Hibiscus leaves

Procedia PDF Downloads 436

Authors: Yung-Feng Yen, I-an Jen, Yi-Ming Arthur Chen

Abstract:

The molecular mimicry between human immunodeficiency virus (HIV) protein and red blood cell (RBC) antigens could induce the production of anti-RBC autoantibodies. However, the association between HIV infection and subsequent development of autoimmune hemolytic anemia (AIHA) remains unclear. This nationwide population-based cohort study aimed to determine the association between incident AIHA and HIV in Taiwan. From 2000–2012, we identified adult people living with HIV/AIDS (PLWHA) from the Taiwan centers for disease control HIV Surveillance System. HIV-infected individuals were defined by positive HIV-1 western blot. Age- and sex-matched controls without HIV infection were selected from the Taiwan National Health Insurance Research Database for comparison. All patients were followed until Dec. 31, 2012, and observed for occurrence of AIHA. Of 171,468 subjects (19,052 PLWHA, 152,416 controls), 30 (0.02%) had incident AIHA during a mean follow-up of 5.45 years, including 23 (0.12%) PLWHA and 7 (0.01%) controls. After adjusting for potential confounders, HIV infection was found to be an independent risk factor of incident AIHA (adjusted hazard ratio [AHR], 20.9; 95% confidence interval [CI], 8.34-52.3). Moreover, PLWHA receiving HAART were more likely to develop AIHA than those not receiving HAART (AHR, 10.8; 95% CI, 2.90-40.1). Additionally, the risk of AIHA was significantly increased in those taking efavirenz (AHR, 3.15; 95% CI, 1.18-8.43) or atazanavir (AHR, 6.58; 95% CI, 1.88-22.9) component of the HAART. In conclusion, HIV infection is an independent risk factor for incident AIHA. Clinicians need to be aware of the higher risk of AIHA in PLWHA.

Keywords: autoimmune disease , hemolytic anemia, HIV, highly active antiretroviral treatment

Procedia PDF Downloads 236

Authors: Jiandong Ren, Lijun Zhao, Peng Chen

Abstract:

Nafamostat (NA), a synthetic broad-spectrum serine protease inhibitor, has been routinely employed for the treatment of acute pancreatitis (AP) and other inflammatory-associated diseases in some East Asia countries. Although the potent inhibitory activity against inflammation-related proteases such as thrombin, trypsin, kallikrein, plasmin, coagulation factors and complement factors is generally considered to be responsible for the anti-inflammatory effects of NA, precise target and molecular mechanism underlying the anti-inflammatory activity in the treatment of AP remain largely unknown yet. As an intracellular inflammatory signaling platform, the NOD-like receptor protein 3 (NLRP3) inflammasome is recently identified to be involved in the development of AP. In present study, we have revealed that NA alleviated pancreatic injury in a caerulein-induced AP model by inhibiting the NLRP3 inflammasome activation in pancreas. Mechanistically, NA interacted with HDAC6, a cytoplasmic deacetylase implicated in the NLRP3 inflammasome pathway, and efficiently abrogated the function of HDAC6. This property enabled NA to influence HDAC6 dependent NF-κB transcriptional activity and thus block NF-κB-driven transcriptional priming of NLRP3 inflammasome. Moreover, NA exerted the potential to interfere HDAC6-mediated intracellular transport of NLRP3, thereby leading to the failure of NLRP3 inflammasome activation. Our current work has provided valuable insight into the molecular mechanism underlying the immunomodulatory effect of NA in treatment of AP, highlighting its promising application in prevention of NLRP3 inflammasome-associated inflammatory pathological damage.

Keywords: acute pancreatitis, HDAC6, nafamostat, NLRP3 inflammasome

Procedia PDF Downloads 72

Authors: Sarah Devi Silvian, Hanna Shobrina Iqomatul Haq, Fara Cholidatun Nabila, Agustin Krisna Wardani

Abstract:

Bacteria ability to survive in antibiotic is controlled by the presence of gene that encodes the antibiotic resistance protein. The instability of the antibiotic resistance gene can be observed by exposing the bacteria under the lethal dose of antibiotic. Low concentration of antibiotic can induce mutation, which may take a role in bacterial adaptation through the antibiotic concentration. Lactobacillus casei FNCC 0090 is one of the probiotic bacteria that has an ability to survive in tetracycline by expressing the tetM gene. The aims of this study are to observe the possibilities of mutation happened in L.casei FNCC 0090 by exposing in sub-lethal dose of tetracycline and also observing the instability of the tetM gene by comparing the sequence between the wild type and mutant. L.casei FNCC 0090 has a lethal dose in 60 µg/ml, low concentration is applied to induce the mutation, the range from 10 µg/ml, 15 µg/ml, 30 µg/ml, 45 µg/ml, and 50 µg/ml. L.casei FNCC 0090 is exposed to the low concentration from lowest to the highest concentration to induce the adaptation. Plasmid is isolated from the highest concentration culture which is 50 µg/ml by using modified alkali lysis method with the addition of lysozyme. The tetM gene is isolated by using PCR (Polymerase Chain Reaction) method, then PCR amplicon is purified and sequenced. Sequencing is done on both samples, wild type and mutant. Both sequences are compared and the mutations can be traced in the presence of nucleotides changes. The changing of the nucleotides means that the tetM gene is instable.

Keywords: L. casei FNCC 0090, probiotic, tetM, tetracycline

Procedia PDF Downloads 191

Authors: Nafiseh Noormohammadi, Ahmad Sobhani Najafabadi

Abstract:

Hypericins (hypericin and pseudohypericin) are natural napthodianthrone derivatives produced by Hypericum perforatum (St. John’s Wort), which have many medicinal properties such as antitumor, antineoplastic, antiviral, and antidepressant activities. Production and accumulation of hypericin in the plant are influenced by both genetic and environmental conditions. Despite the existence of different high-throughput data on the plant, genetic dimensions of hypericin biosynthesis have not yet been completely understood. In this research, 21 high-quality RNA-seq data on different parts of the plant were integrated into metabolic data to reconstruct a coexpression network. Results showed that a cluster of 30 transcripts was correlated with total hypericin. The identified transcripts were divided into three main groups based on their functions, including hypericin biosynthesis genes, transporters, detoxification genes, and transcription factors (TFs). In the biosynthetic group, different isoforms of polyketide synthase (PKSs) and phenolic oxidative coupling proteins (POCPs) were identified. Phylogenetic analysis of protein sequences integrated into gene expression analysis showed that some of the POCPs seem to be very important in the biosynthetic pathway of hypericin. In the TFs group, six TFs were correlated with total hypericin. qPCR analysis of these six TFs confirmed that three of them were highly correlated. The identified genes in this research are a rich resource for further studies on the molecular breeding of H. perforatum in order to obtain varieties with high hypericin production.

Keywords: hypericin, St. John’s Wort, data mining, transcription factors, secondary metabolites

Procedia PDF Downloads 94

Authors: S. Rajeswari

Abstract:

Medicinal Plant extracts and their bioactive compounds have been used for antimicrobial activities and have significant remedial properties. In the recent years, a wide range of investigations have been carried out throughout the world to confirm antimicrobial properties of different medicinally important plants. A number of plants showed efficient antimicrobial activities, which were comparable to that of synthetic standard drugs or antimicrobial agents. The large family Euphorbiaceae contains nearly about 300 genera and 7,500 speciesand one among is Ricinus communis or castor plant which has high traditional and medicinal value for disease free healthy life. Traditionally the plant is used as laxative, purgative, fertilizer and fungicide etc. whereas the plant possess beneficial effects such as anti-oxidant, antihistamine, antinociceptive, antiasthmatic, antiulcer, immunomodulatory anti diabetic, hepatoprotective, anti inflammatory, antimicrobial, and many other medicinal properties. This activity of the plant possess due to the important phytochemical constituents like flavonoids, saponins, glycosides, alkaloids and steroids. The presents study includes the phytochemical properties of Ricinus communis and to prediction of the anti-microbial activity of Ricinus communis using DNA gyrase of Staphylococcus aureus as molecular target. Docking results of varies chemicals compounds of Ricinus communis against DNA gyrase of Staphylococcus aureus by maestro 9.8 of Schrodinger show that the phytochemicals are effective against the target protein DNA gyrase. our studies suggest that the phytochemical from Ricinus communis such has INDICAN (G.Score 4.98) and SUPLOPIN-2(G.Score 5.74) can be used as lead molecule against Staphylococcus infections.

Keywords: euphorbiaceae, antimicrobial activity, Ricinus communis, Staphylococcus aureus

Procedia PDF Downloads 480

Authors: Suthida Panwong, Jeeraporn Pekkoh, Yingmanee Tragoolpua, Aussara Panya

Abstract:

Dengue virus (DENV) infection is a major public health problem in many countries, especially in tropical and subtropical countries. DENV infection causes dengue fever that can progress to serious conditions of dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS), relevant to a high risk of mortality. However, there are no effective treatments available against the manifestation and fatalities. Currently, natural extracts have been widely used for the treatment of infectious diseases due to their safety, non-accumulation in the body, or lower side effects. Chlorella spp. is a microalgae with anti-viral activity, but there is not much report to support its ability to inhibit DENV infection. Thus, this study aimed to investigate the inhibitory effect of ethanolic extract from Chlorella sp. AARL G049, which was explored in Thailand on inhibition of DENV-2 infection. The inhibitory effect on viral infection was assessed using a foci-forming assay (FFA), which revealed that a concentration of 125 µg/mL could inhibit viral infection in Vero cells by 75.45±8.06% when treated at the same time as DENV-2 infection. Moreover, the extract at an equal concentration effectively reduced viral protein synthesis by 90.51±5.48% when assessed in human cell lines using enzyme-linked immunosorbent assay (ELISA). Concordantly, the number of infected cells after treatment was reduced as measured by immunofluorescent assay (IFA). Therefore, the finding of this study supports the potential use of Chlorella sp. extract to suppress DENV infection.

Keywords: viral infection, flavivirus, treatment, natural extract

Procedia PDF Downloads 32

Authors: Muhammad Ihsan Andi Dagong, Cece Sumantri, Ronny Rachman Noor, Rachmat Herman, Mohamad Yamin

Abstract:

Calpastatin involved in various physiological processes in the body such as the protein turnover, growth, fusion and mioblast migration. Thus, allegedly Calpastatin gene diversity (CAST) have an association with growth and potential use as candidate genes for growth trait. This study aims to identify the association between the genetic diversity of CAST gene with some growth properties such as body dimention (morphometric), body weight and daily weight gain in sheep. A total of 157 heads of Thin Tail Sheep (TTS) reared intensively for fattening purposes in the uniform environmental conditions. Overall sheep used were male, and maintained for 3 months. The parameters of growth properties were measured among others: body weight gain (ADG) (g/head / day), body weight (kg), body length (cm), chest circumference (cm), height (cm). All the sheep were genotyped by using PCR-SSCP (single strand conformational polymorphism) methods. CAST gene in locus fragment intron 5 - exon 6 were amplified with a predicted length of about 254 bp PCR products. Then the sheep were stratified based on their CAST genotypes. The result of this research showed that no association were found between the CAST gene variations with morphometric body weight, but there was a significant association with daily body weight gain (ADG) in sheep observed. CAST-23 and CAST-33 genotypes has higher average daily gain than other genotypes. CAST-23 and CAST-33 genotypes that carrying the CAST-2 and CAST-3 alleles potential to be used in the selection of the nature of the growth trait of the TTS sheep.

Keywords: body weight, calpastatin, genotype, growth trait, thin tail sheep

Procedia PDF Downloads 322

Authors: Deepika Biswas

Abstract:

Acinetobacter baumannii, a hospital-acquired pathogen, causes nosocomial infections including pneumonia, urinary tract infection, and secondary meningitis. Carbapenem is most effective antibiotics against it. Its increased resistance to carbapenems has been a rising global concern. Antibiotics such as carbapenem are unable to use on hospital setups to eradicate A. baumannii, hence different concentrations of disinfectants including phenol; sodium hypochlorite and ethanol are increasingly being used. The objective of the present study is to find an effective concentration of above disinfectants against carbapenem-resistant strain RS307 of A. baumannii. Growth kinetics of RS307 has been determined using UV-Vis spectrophotometer in the presence and absence of disinfectants in triplicate and its standard deviation has also been calculated which make the results more reliable. Differential growth curves were plotted, which showed the effective concentration among all the concentrations of phenol, sodium hypochlorite and ethanol. On disc diffusion assay, antimicrobial effect was observed by comparing all the concentrations of disinfectants to check its synergy with imipenem, most effective carbapenem. All the results collectively revealed that 0.5% phenol, 0.5% sodium hypochlorite, and 70% ethanol could preferably be used as disinfectant for hospital setup against the carbapenem-resistant strain of A. baumannii. SDS PAGE analysis showed differential expression in the protein profile of A. baumannii after treatment. The present study highlighted that few disinfectants even in low concentration had shown better antimicrobial activity hence may be recommended for regular use in the hospitals, which will be cost effective and less harmful.

Keywords: Acenatobacter bomunii, phenol, sodium hypoclirite, ethanol, carbapenem resistance, disinfectant

Procedia PDF Downloads 258

Authors: Walaa Mohammed Saeed

Abstract:

Studies suggest a crosstalk between bone and metabolism through Osteocalcin (OC), a bone-derived protein that plays an important role in regulating glucose and fat metabolism. Studies relate type II Diabetes Mellitus (DMII) with Homocysteine (Hcy) and cardiovascular diseases (CVD). This study investigates the relationship between levels of OC, Hcy, and DMII in 85 subjects of which 50 were diabetic female patients (29–65 years) and 35 healthy controls. OC and Hcy levels were measured in fasting blood samples using immunoassay analyzer. Fasting serum glucose, glycated hemoglobin, lipid profile, were estimated by automated Siemens Dimension XP auto-analyzer. A significant increase in the frequency of low OC levels (p < 0.001) and high Hcy levels (p < 0.001) was detected in diabetic patients compared to controls (chi-squared test). Using ANOVA test, patients were divided into tertiles based on plasma OC and Hcy levels; fasting serum glucose varied inversely with OC but directly with Hcy tertiles (p=0.049, p=0.033 respectively). Atherogenic Index of Plasma (AIP=Log TG/HDL) predicts that diabetic patients with 36% high and 15% intermediate cardiovascular risk had increased frequency of low OC levels compared to low-risk patients (p=0.047). Another group of diabetic patients with 39% high and 11% intermediate CVD risk had increased frequency of high Hcy levels (p=0.033). A significant negative correlation existed between OC and glucose (r = -0.318; p = 0.035) while correlation between glucose level and Hcy (r = 0.851 p=0.022) was positive. Hence, low serum OC levels and high Hcy levels were associated with impaired glucose metabolism that may increase cardiovascular risk in DMII.

Keywords: osteocalcin, homocysteine, type 2 diabetes, cardiovascular

Procedia PDF Downloads 155

Authors: Gian Carlo F. Maliwat, Stephanie F. Velasquez, Janice A. Ragaza

Abstract:

A 50-day growth trial was conducted to evaluate the efficacy of Chlorella vulgaris (Beijerinck) as an ingredient in the diets of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae (PL30). Immune response (total haemocyte count and prophenoloxidase activity) was also assessed by subjecting postlarvae to a challenge test against Aeromonas hydrophila (Chester) for 14 days. Isonitrogenous and iso-lipidic test diets were prepared using a fishmeal-based-positive control diet (D0) and four basal diets with inclusion levels of 2% (D2), 4% (D4), 6% (D6) and 8% (D8) C. vulgaris. Postlarvae of M. rosenbergii were randomly stocked (mean initial body weight of 0.19 ± 0.02 g) in 30-L tanks in three replicates per dietary treatment for evaluation of growth performance. Another set of postlarvae (mean initial body weight of 1.25 ± 0.02 g) was randomly distributed in 95-L tanks in three replicates per dietary treatment for the assessment of immune response. Results showed that specific growth rate was significantly higher (P < 0.05) in postlarvae fed D4 and D6. Variations in values for carcass protein, lipid, moisture, and ash were also evident. Postlarvae fed diets with Chlorella showed increased prophenol oxidase activity and total haemocyte counts. Moreover, the survival rate after challenge with A. hydrophila was significantly increased (P < 0.05). Inclusion of C. vulgaris in diets enhanced immune response and resistance of M. rosenbergii postlarvae against A. hydrophila infection.

Keywords: Chlorella vulgaris, haemocyte count, Macrobrachium rosenbergii, prophenoloxidase activity

Procedia PDF Downloads 154

Authors: Bhupinder Kaur, P. P. Srivastav

Abstract:

The fruit and vegetable industries are responsible for producing huge amount of waste, which is a problem to environmental safety and should be utilized efficiently. Mango (Mangifera indica L.) is an important commercially grown fruit and referred as the “King of fruits”. In 2015, India was the largest producer (18.506 MT) of mangoes and out of which 9.16 % lost during post-harvest handling. The mango kernel and peel represent approximately 17-22% and 7-22% of the overall mass of fruit respectively and discarded as waste. Hence, an attempt has been made with three mango cultivars (Langra, Dashehari, Fazli) to investigate the effect of cryogenic grinding on various characteristics of mango peel powder (MPP). The cryogenic grinding is an emerging technology which is used for retention of beneficial volatile and bioactive components. The feed rate was highest for Langra followed by Chausa. The samples have 2-4% fat along with significant amount of protein (4-6%) and crude fiber (9-13%). Mango peel is also a good source of minerals such as calcium, potassium, manganese, iron, copper, zinc, and magnesium. Interestingly, the significant amount of essential minerals like phosphorus and chlorine in all the varieties was found with the highest value in Langra (phosphorus 10.83% and chlorine 2.41%) which are not reported earlier. SEM analysis revealed the surface morphology and shape of the particles. Waste utilization is a promising measure from both an environmental and economic point of view. Chemical characterization of the samples indicated its potential to be used for the fortification of food products which in turn reduces hazards due to waste and improve functional quality of the foods.

Keywords: cryogenic grinding, morphological, mineral composition, SEM

Procedia PDF Downloads 236

Authors: Rohit Sane, Pravin Ghadigaonkar, Purvi Ahuja, Suvarna Tirmare, Archana Kelhe, Kranti Shinde, Rahul Mandole

Abstract:

To evaluate the efficacy of Comprehensive Diabetic Care Program with the reduction of HbA1c in overweight Diabetes Mellitus Type II patients retrospectively. Methods: Retrospective study was carried out on 34 overweight type II diabetic patients (Mean Age = 54.58 ±11.38 yrs). A total of 34 patients were enrolled after screening of 68 patients (HbA1c 7-10%). The patients were on concomitant drugs namely insulin (11.76%), DPP-4 inhibitor (17.64%), Biguanide (55.88%), Sulfonylurea (52.94%), thiazolidinedione (11.76%), other medications (20.58%) and no allopathic medications (14.70%). The patients were given Comprehensive Diabetic Care Program consisting of panchkarma procedures namely snehana (external oleation), swedana (passive heat therapy) and basti (enema), which was completed in 15 sittings. During the therapy and next 90 days, the patients followed low carbohydrate and moderate protein & fat diet. The primary endpoint of this study was the evaluation of reduction in HbA1c at the end of the follow-up after 90 days. Results: Thirty-four overweight type II diabetic patients (mean age: 54.58[±11.38], HbA1c[7-10%], 67.64% male and 32.35% female) were enrolled in the study. A significant reduction was observed in HbA1c levels (14.30%, p<0.05) at the end of the 90 days follow-up as compared to baseline. Also, BMI was reduced by 5.87%. There was reduction in the usage of the concomitant drugs namely insulin (2.94%), DPP-4 inhibitor (2.94%), Biguanide (32.35%), Sulfonylurea (35.29%), thiazolidinedione (5.88%), other medications(17.64%) and no allopathic medications (32.35%). Conclusion: The results of the study highlight not only in the reduction of HbA1c, but also in BMI and drug tapering of the CDC program in the overweight type II diabetic patients with HbA1c (7-10%).

Keywords: HbA1c, low carb diet, Panchakarma therapy, Type II Diabetes

Procedia PDF Downloads 284

Authors: Shukla Ila, Azmi Lubna, S. S. Gupta, Ch. V. Rao

Abstract:

Oxidative stress has been increasingly associated with the induction and progression of liver damage. The current study was conducted to record the effect of combination of Lactobacillus and Lichen rangiferinus extract (LRE + Lac) on the severity of injury in experimental alcoholic liver disease and how it affects plasma levels of prostaglandin E2, endotoxin, thromboxane B2, and leukotriene B4. Male Wistar rats were grouped into five comprising six animals in each group. Group 1 served as negative control. Groups 2-5 were administered 10% ethanol for six weeks. Group 3 was administered with extract (200 mg/kg), group 4 received the diet containing 10% ethanol plus a bolus of lactobacilli GG (1010 CFU), and group 5 animals were given silymarin along with alcohol and it served as positive control. Aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total protein content, γ-glutamyltransferase, glutathione S-transferase, oxidative stress markers, glutathione, malondialdehyde and glutathione reductase were determined using standard diagnostic kits. Histopathological analysis of liver tissue was also made. A positive relation was found between plasma endotoxin levels and degree of liver injury. The pathology records were also related positively with leukotriene B4 and thromboxane B2. But a negative correlation was obtained with PgE2 levels. This study led us to hypothesize that the increased endotoxin levels modulate liver metabolism of eicosanoid, which gradually leads to liver injury. Endotoxemia increases leukotriene and thromboxane levels in plasma.

Keywords: lactobacillus, Lichen rangiferinus, endotoxemia, silymarin

Procedia PDF Downloads 325

Authors: Vun Yee Thien, Kenneth Francis Rodrigues, Clemente Michael Vui Ling Wong, Wilson Thau Lym Yong

Abstract:

Transcriptomes associated with the process of photosynthesis have offered insights into the mechanism of gene regulation in terrestrial plants; however, limited information is available as far as macroalgae are concerned. This investigation aims to decipher the underlying mechanisms associated with photosynthesis in the red alga, Kappaphycus alvarezii, by performing a differential expression analysis on a de novo assembled transcriptomes. Comparative analysis of gene expression was designed to examine the alteration of light qualities and its effect on physiological mechanisms in the red alga. High-throughput paired-end RNA-sequencing was applied to profile the transcriptome of K. alvarezii irradiated with different wavelengths of light (blue 492-455 nm, green 577-492 nm and red 780-622 nm) as compared to the full light spectrum, resulted in more than 60 million reads individually and assembled using Trinity and SOAPdenovo-Trans. The transcripts were annotated in the NCBI non-redundant (nr) protein, SwissProt, KEGG and COG databases with a cutoff E-value of 1e-5 and nearly 30% of transcripts were assigned to functional annotation by Blast searches. Differential expression analysis was performed using edgeR. The DEGs were designated to six categories: BL (blue light) regulated, GL (green light) regulated, RL (red light) regulated, BL or GL regulated, BL or RL regulated, GL or RL regulated, and either BL, GL or RL regulated. These DEGs were mapped to terms in KEGG database and compared with the whole transcriptome background to search for genes that regulated by light quality. The outcomes of this study will enhance our understanding of molecular mechanisms underlying light-induced responses in red algae.

Keywords: de novo transcriptome sequencing, differential gene expression, Kappaphycus alvareziired, red alga

Procedia PDF Downloads 509

Authors: Ga-Young Lee, Hyun-Man Kim

Abstract:

The modulation of the cell-cell junction is critical in epithelial-mesenchymal transition during tumorigenesis. As RhoA activity is known to be up-regulated to dissociate cell-cell junction by contracting acto-myosin complex in various cancer cells, the present study investigated if RhoA activity was also associated with the disruption of the cell-cell junction of oral cancer cells. We studied SCC-25 cells which are established from oral squamous cell carcinoma if their E-cadherin junction (ECJ) was under control of RhoA. Interestingly, development of ECJ of SCC-25 cells depended on the amount of fibronectin (FN) coated on the culture dishes. Seeded cells promptly aggregated to develop ECJ on the substrates coated with a low amount of FN, whereas they were retarded in the development of ECJ on the substrates coated with a high amount of FN. However, it was an unexpected finding that total RhoA activity was lower in the dissociated cells on the substrates of high FN than in the aggregated cells on the substrates of low FN. Treating the dissociated cells on the substrates of high FN with LPA, a RhoA activator, promoted the development to ECJ. In contrast, treating the aggregated cells on the substrates of low FN with Clostridium botulinum C3, a toxin decreasing RhoA activity, dissociated cells concomitant with the disruption of ECJ. Genetical knockdown of RhoA expression by transfecting RhoA siRNA also down-regulated the development of ECJ in SCC-25 cells. Furthermore, PMA, an activator of protein kinase C (PKC), down-regulated the development of ECJ junction of SCC-25 cells on the substrates coated with low FN. In contrast, GO6976, a PKC inhibitor, up-regulated the development of ECJ of SCC-25 cells with the activation of RhoA on the substrates coated with high FN. In conclusion, in the present study, we demonstrated unexpected results that the activation of RhoA promotes the development of ECJ, whereas the inhibition of RhoA retards the development of ECJ in SCC-25 cells.

Keywords: E-cadherin junction, oral squamous cell carcinoma, PKC, RhoA, SCC-25

Procedia PDF Downloads 332

Authors: Asma Alhuqail, Nagwa Aref

Abstract:

Programmed cell death resembles a real nature active defense in Datura metel against TMV after three days of virus infection. Physiological plant response was assessed for asymptomatic healthy and symptomatic infected detached leaves. The results indicated H2O2 and Chlorophyll-a as the most potential parameters. Chlorophyll-a was considered the only significant predictor variant for the H2O2 dependent variant with a P value of 0.001 and R-square of 0.900. The plant immune response was measured within three days of virus infection using the cutoff value of H2O2 (61.095 lmol/100 mg) and (63.201 units) for the tail moment in the Comet Assay. Their percentage changes were 255.12% and 522.40% respectively which reflects the stress of virus infection in the plant. Moreover, H2O2 showed 100% specificity and sensitivity in the symptomatic infected group using the receiver-operating characteristic (ROC). All tested parameters in the symptomatic infected group had significant correlations with twenty-five positive and thirty-one negative correlations where the P value was <0.05 and 0.01. Chlorophyll-a parameter had a crucial role of highly significant correlation between total protein and salicylic acid. Contrarily, this correlation with tail moment unit was (r = _0.930, P <0.01) where the P value was < 0.01. The strongest significant negative correlation was between Chlorophyll-a and H2O2 at P < 0.01, while moderate negative significant correlation was seen for Chlorophyll-b where the P value < 0.05. The present study discloses the secret of the three days of rapid transient production of activated oxygen species (AOS) that was enough for having potential quantitative physiological parameters for defensive plant response toward the virus.

Keywords: programmed cell death, plant–adaptive immune response, hydrogen peroxide (H2O2), physiological parameters

Procedia PDF Downloads 248

Authors: Ahmed Alkarimi, Kevin Welham

Abstract:

Organic mixed mode monolithic columns were fabricated from; glycidyl methacrylate-co-ethylene dimethacrylate-co-stearyl methacrylate, using glycidyl methacrylate and stearyl methacrylate as co monomers representing 30% and 70% respectively of the liquid volume with ethylene dimethacrylate crosslinker and 2,2-dimethoxy-2-phenylacetophenone as the free radical initiator. The monomers were mixed with a binary porogenic solvent, comprising propan-1-ol, and methanol (0.825 mL each). The monolith was formed by photo polymerization (365 nm) inside a borosilicate glass tube (1.5 mm ID and 3 mm OD x 50 mm length). The monolith was observed to have formed correctly by optical examination and generated reasonable backpressure, approximately 650 psi at a flow rate of 0.2 mL min⁻¹ 50:50 acetonitrile: water. The morphological properties of the monolithic columns were investigated using scanning electron microscopy images, and Brunauer-Emmett-Teller analysis, the results showed that the monolith was formed properly with 19.98 ± 0.01 mm² surface area, 0.0205 ± 0.01 cm³ g⁻¹ pore volume and 6.93 ± 0.01 nm average pore size. The polymer monolith formed was further investigated using proton nuclear magnetic resonance, and Fourier transform infrared spectroscopy. The monolithic columns were investigated using high-performance liquid chromatography to test their ability to separate different samples with a range of properties. The columns displayed both hydrophobic/hydrophilic and hydrophobic/ion exchange interactions with the compounds tested indicating that true mixed mode separations. The mixed mode monolithic columns exhibited significant separation of proteins.

Keywords: LC separation, proteins separation, monolithic column, mixed mode

Procedia PDF Downloads 164