Search results for: enzyme linked immunosorbent assay (ELISA)
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3365

Search results for: enzyme linked immunosorbent assay (ELISA)

3335 Chemical Synthesis of a cDNA and Its Expression Analysis

Authors: Salman Akrokayan

Abstract:

Synthetic cDNA (ScDNA) of granulocyte colony-stimulating factor (G-CSF) was constructed using a DNA synthesizer with the aim to increase its expression level. 5' end of the ScDNA of G-CSF coding region was modified by decreasing the GC content without altering the predicted amino acids sequence. The identity of the resulting protein from ScDNA was confirmed by the highly specific enzyme-linked immunosorbent assay. In conclusion, a synthetic G-CSF cDNA in combination with the recombinant DNA protocol offers a rapid and reliable strategy for synthesizing the target protein. However, the commercial utilization of this methodology requires rigorous validation and quality control.

Keywords: synthetic cDNA, recombinant G-CSF, cloning, gene expression

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3334 Surveillance for African Swine Fever and Classical Swine Fever in Benue State, Nigeria

Authors: A. Asambe, A. K. B. Sackey, L. B. Tekdek

Abstract:

A serosurveillance study was conducted to detect the presence of antibodies to African swine fever virus (ASFV) and Classical swine fever virus in pigs sampled from piggeries and Makurdi central slaughter slab in Benue State, Nigeria. 416 pigs from 74 piggeries across 12 LGAs and 44 pigs at the Makurdi central slaughter slab were sampled for serum. The sera collected were analysed using Indirect Enzyme Linked Immunosorbent Assay (ELISA) test kit to test for antibodies to ASFV, while competitive ELISA test kit was used to test for antibodies to CSFV. Of the 416 pigs from piggeries and 44 pigs sampled from the slaughter slab, seven (1.7%) and six (13.6%), respectively, tested positive to ASFV antibodies and was significantly associated (p < 0.0001). Out of the 12 LGAs sampled, Obi LGA had the highest ASFV antibody detection rate of (4.8%) and was significantly associated (p < 0.0001). None of the samples tested positive to CSFV antibodies. The study concluded that antibodies to CSFV were absent in the sampled pigs in piggeries and at the Makurdi central slaughter slab in Benue State, while antibodies to ASFV were present in both locations; hence, the need to keep an eye open for CSF too since both diseases may pose great risk in the study area. Further studies to characterise the ASFV circulating in Benue State and investigate the possible sources is recommended. Routine surveillance to provide a comprehensive and readily accessible data base to plan for the prevention of any fulminating outbreak is also recommended.

Keywords: African swine fever, classical swine fever, piggery, slaughter slab, surveillance

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3333 Effects of Probiotics on Specific Immunity in Broiler Chicken in Syria

Authors: Moussa Majed, Omar Yaser

Abstract:

The main objective of this experiment was to study the impact of Probiotic compound on the specific immunity as the case study of infectious bursal disease. Total of 8000 one-day old Ross 108 broiler were randomly divided into two experimental groups; control group (4500 birds) and experimental group (3500 birds). Birds in two groups were reared under similar environmental conditions. Birds in control group received basal diets without probiotic whereas the birds in experimental one were fed basal diets supplemented with a commercial probiotic mixture) probiotic lacting k, which contains bacteria cells beyond to lactobacillus, Streptococcus and bifidobacterium genus that are isolated from gut microflora in healthy chickens(. The commercial probiotic were used according to the manufacturer instruction. 400 blood samples for each group were collected from wing vein every 5-7 days as interval period till 42 days old. Indirect Enzyme-Linked Immunosorbent Assay (ELISA) test was performed to detect the level of infectious bursal disease virus (IBDV) antibodies. The results clearly showed that the mean of immune titers was significantly (p= 0.03) higher in trail group than control one. The coefficient of variance percentages were 55% and 39% for control and trial groups respectively, this illustrates that homogeneity of immunity titers in the trail group was much better comparing with control group. The values of geometric means of titers in the control group and trial group were reported 3820 and 8133, respectively. The crude mortality rate in the experimental group was two times lower comparing with control group (14% and 28% respectively, p = 0.005

Keywords: probiotic, broiler chicken, infectious bursal disease, immunity, ELISA test

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3332 The Pathology of Bovine Rotavirus Infection in Calves That Confirmed by Enzyme Linked Immunosorbant Assay, Reverse Transcription Polymerase Chain Reaction and Real-Time RT-PCR

Authors: Shama Ranjan Barua, Tofazzal M. Rakib, Mohammad Alamgir Hossain, Tania Ferdushy, Sharmin Chowdhury

Abstract:

Rotavirus is one of the main etiologies of neonatal diarrhea in bovine calves that causes significant economic loss in Bangladesh. The present study was carried out to investigate the pathology of neonatal enteritis in calves due to bovine rotavirus infection in south-eastern part of Bangladesh. Rotavirus was identified by using ELISA, RT-PCR (Reverse Transcription Polymerase Chain Reaction), real-time RT-PCR. We examined 12 dead calves with history of diarrhea during necropsy. Among 12 dead calves, in gross examination, 6 were found with pathological changes in intestine, 5 calves had congestion of small intestine and rest one had no distinct pathological changes. Intestinal contents and/or faecal samples of all dead calves were collected and examined to confirm the presence of bovine rotavirus A using Enzyme linked immunosorbant assay (ELISA), RT-PCR and real-time RT-PCR. Out 12 samples, 5 (42%) samples revealed presence of bovine rotavirus A in three diagnostic tests. The histopathological changes were found almost exclusively limited in the small intestine. The lesions of rotaviral enteritis ranged from slight to moderate shortening (atrophy) of villi in the jejunum and ileum with necrotic crypts. The villi were blunt and covered by immature epithelial cells. Infected cells, stained with Haematoxylin and Eosin staining method, showed characteristic syncytia and eosinophilc intracytoplasmic inclusion body. The presence of intracytoplasmic inclusion bodies in enterocytes is the indication of viral etiology. The presence of rotavirus in the affected tissues and/or lesions was confirmed by three different immunological and molecular tests. The findings of histopathological changes will be helpful in future diagnosis of rotaviral infection in dead calves.

Keywords: calves, diarrhea, pathology, rotavirus

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3331 Grains of Winter Wheat Spelt (Triticum spelta L.) for Save Food Production

Authors: D. Jablonskytė-Raščė, A. Mankevičienė, S. Supronienė, I. Kerienė, S. Maikštėnienė, S. Bliznikas, R. Česnulevičienė

Abstract:

Organic farming does not allow the use of conventional mineral fertilizers and crop protection products. As a result, in our experiments we chose to grow different species of cereals and to see how cereal species affects mycotoxin accumulation. From the phytopathological and entomological viewpoint, the glumes of spelt grain perform a positive role since they protect grain from the infection of pathogenic microorganisms. On the background of the above-mentioned infection, there were more Fusarium–affected grains of spelt than of common wheat. It can be assumed that spelt is more susceptible to the Fusarium fungi infection than common wheat. This study describes the occurrence of DON, ZEA and T2/HT2 toxin in a survey of spelt and common wheat and their bran as well as flour. The analysis was conducted using the enzyme-linked immunosorbent assay (ELISA) method. The concentrations of DON, ZEA, and T2/HT2 in Triticum spelta and Triticum aestivum are influenced by species, cereal type and year interaction. The highest concentration of mycotoxin was found in spelt grain with glumes. The obtained results indicate the significantly higher concentrations of Fusarium toxins in glumes than in dehulled grain which implicate the possible protective effect of spelt wheat glumes. The lowest DON, ZEA, and T2/HT2 concentration was determined in spelt grain without glumes.

Keywords: Fusarium mycotoxins, organic farming, spelt

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3330 Development of a Novel Score for Early Detection of Hepatocellular Carcinoma in Patients with Hepatitis C Virus

Authors: Hatem A. El-Mezayen, Hossam Darwesh

Abstract:

Background/Aim: Hepatocellular carcinoma (HCC) is often diagnosed at advanced stage where effective therapies are lacking. Identification of new scoring system is needed to discriminate HCC patients from those with chronic liver disease. Based on the link between vascular endothelial growth factor (VEGF) and HCC progression, we aimed to develop a novel score based on combination of VEGF and routine laboratory tests for early prediction of HCC. Methods: VEGF was assayed for HCC group (123), liver cirrhosis group (210) and control group (50) by Enzyme Linked Immunosorbent Assay (ELISA). Data from all groups were retrospectively analyzed including α feto protein (AFP), international normalized ratio (INR), albumin and platelet count, transaminases, and age. Areas under ROC curve were used to develop the score. Results: A novel index named hepatocellular carcinoma-vascular endothelial growth factor score (HCC-VEGF score)=1.26 (numerical constant) + 0.05 ×AFP (U L-1)+0.038 × VEGF(ng ml-1)+0.004× INR –1.02 × Albumin (g l-1)–0.002 × Platelet count × 109 l-1 was developed. HCC-VEGF score produce area under ROC curve of 0.98 for discriminating HCC patients from liver cirrhosis with sensitivity of 91% and specificity of 82% at cut-off 4.4 (ie less than 4.4 considered cirrhosis and greater than 4.4 considered HCC). Conclusion: Hepatocellular carcinoma-VEGF score could replace AFP in HCC screening and follow up of cirrhotic patients.

Keywords: Hepatocellular carcinoma, cirrhosis, HCV, diagnosis, tumor markers

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3329 Diagnostic Performance of Tumor Associated Trypsin Inhibitor in Early Detection of Hepatocellular Carcinoma in Patients with Hepatitis C Virus

Authors: Aml M. El-Sharkawy, Hossam M. Darwesh

Abstract:

Abstract— Background/Aim: Hepatocellular carcinoma (HCC) is often diagnosed at advanced stage where effective therapies are lacking. Identification of new scoring system is needed to discriminate HCC patients from those with chronic liver disease. Based on the link between tumor associated trypsin inhibitor (TATI) and HCC progression, we aimed to develop a novel score based on combination of TATI and routine laboratory tests for early prediction of HCC. Methods: TATI was assayed for HCC group (123), liver cirrhosis group (210) and control group (50) by Enzyme Linked Immunosorbent Assay (ELISA). Data from all groups were retrospectively analyzed including α feto protein (AFP), international normalized ratio (INR), albumin and platelet count, transaminases, and age. Areas under ROC curve were used to develop the score. Results: A novel index named hepatocellular carcinoma-vascular endothelial growth factor score (HCC-TATI score) = 3.1 (numerical constant) + 0.09 ×AFP (U L-1) + 0.067 × TATI (ng ml-1) + 0.16 × INR – 1.17 × Albumin (g l-1) – 0.032 × Platelet count × 109 l-1 was developed. HCC-TATI score produce area under ROC curve of 0.98 for discriminating HCC patients from liver cirrhosis with sensitivity of 91% and specificity of 82% at cut-off 6.5 (ie less than 6.5 considered cirrhosis and greater than 4.4 considered HCC). Conclusion: Hepatocellular carcinoma-TATI score could replace AFP in HCC screening and follow up of cirrhotic patients.

Keywords: Hepatocellular carcinoma, cirrhosis, HCV, diagnosis, TATI

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3328 Urinary Neutrophil Gelatinase Associated Lipocalin as Diagnostic Biomarkers for Lupus Nephritis

Authors: Lorena GóMez Escorcia, Gustavo Aroca MartíNez, Jose Luiz Villarreal, Elkin Navarro Quiroz

Abstract:

Lupus nephritis (LN) is a high-cost disease, occurring in about half of patients with Systemic Lupus Erythematosus (SLE). Renal biopsy constitutes the only protocol that, to date, allows a correct diagnosis of the level of renal involvement in these patients. However, this procedure can have various adverse effects such as kidney bleeding, muscle bleeding, infection, pain, among others. Therefore, the development of new diagnostic alternatives is required. The neutrophil gelatinase-associated lipocalin (NGAL) has been emerging as a novel biomarker of acute kidney injury. The aim of this study was to assess urinary NGAL levels as a marker for disease activity in patients with lupus nephritis. For this work included 50 systemic lupus erythematosus (SLE) patients, 50 with active lupus nephritis (LN), and 50 without autoimmune and renal disease as controls. TNGAL in urine samples was measured by enzyme-linked immunosorbent assay (ELISA). The results revealed that patients with kidney damage had an elevated urinary NGAL as compared to patients with lupus without kidney damage and controls (p <0.005), and the mean of uNGAL was (28.72 ± 4.53), (19.51 ± 4.72), (8.91 ± 3.37) respectively. Measurement of urinary NGAL levels showed a very good diagnostic performance for discriminating patients with Lupus nephritis from SLE without renal damage and of control individuals.

Keywords: lupus nephritis, biomarker, NGAL, urine samples

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3327 The Effect of Malaria Parasitaemia on Serum Reproductive Hormonal Levels of Asymptomatic HIV Subjects in Nauth Nnewi, South Eastern Nigeria

Authors: Ezeugwunne Ifeoma Priscilla, Charles Chinedum Onyenekwe, Joseph Eberendu Ahaneku, Rosemary Adanma Analike, Adesuwa Peace Eidangbe

Abstract:

This study was designed to assess the effect of malaria parasitaemia on serum reproductive hormone levels of asymptomatic HIV adult subjects. A total of 271 participants aged between 17 and 58 ears were conveniently recruited. 135 asymptomatic HIV-infected subjects participated in the study; 67 of them had malaria parasitaemia. 136 HIV seropositive control subjects, 68 of them had malaria parasitaemia. Blood samples were collected from the participants for the determination of HIV status by immunoassay and immunochromatography. Enzyme-linked immunosorbent assay (ELISA) was used to assay for serum LH, FSH, Estrogen, testosterone, progesterone, prolactin, and PSA levels, CD4+T cell counts by Cyflow method, thick and thin films determination of malaria parasitaemia count and density by WHO. Student's t-tests and ANOVA were used to compare means. P<0.05 was considered statistically significant. The results showed significant differences in serum levels of LH, FSH, PSA, estrogen, progesterone, and testosterone amongst the groups at P<0.05, respectively. The serum levels of LH, FSH, and PSA were significantly higher in malaria-infected asymptomatic HIV subjects than in asymptomatic HIV subjects with malaria parasitaemia (P<0.05 in each case). Also, the serum levels of LH, FSH, PSA, estrogen, and progesterone were significantly higher in malaria-infected asymptomatic HIV subjects compared with malaria-infected HIV seronegative subjects (P<0.05, respectively). The mean MP counts and MP density were significantly higher in asymptomatic HIV subjects compared to HIV seronegative subjects (P<0.05, in each case). The mean serum levels of testosterone were significantly lower in both malaria-infected and malaria uninfected HIV seronegative subjects (P<0.05, in each case). In conclusion, Malaria and HIV co-infection might increase the burden of hypogonadism as well as primary testicular failure, hyperprogesteronaemia, elevated levels of estrogen, and PSA in adult males asymptomatic HIV subjects.

Keywords: malaria parasitaemia, HIV, CD4, reproductive hormones

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3326 Chemiluminescent Detection of Microorganisms in Food/Drug Product Using Reducing Agents and Gold Nanoplates

Authors: Minh-Phuong Ngoc Bui, Abdennour Abbas

Abstract:

Microbial spoilage of food/drug has been a constant nuisance and an unavoidable problem throughout history that affects food/drug quality and safety in a variety of ways. A simple and rapid test of fungi and bacteria in food/drugs and environmental clinical samples is essential for proper management of contamination. A number of different techniques have been developed for detection and enumeration of foodborne microorganism including plate counting, enzyme-linked immunosorbent assay (ELISA), polymer chain reaction (PCR), nucleic acid sensor, electrical and microscopy methods. However, the significant drawbacks of these techniques are highly demand of operation skills and the time and cost involved. In this report, we introduce a rapid method for detection of bacteria and fungi in food/drug products using a specific interaction between a reducing agent (tris(2-carboxylethyl)phosphine (TCEP)) and the microbial surface proteins. The chemical reaction was transferred to a transduction system using gold nanoplates-enhanced chemiluminescence. We have optimized our nanoplates synthetic conditions, characterized the chemiluminescence parameters and optimized conditions for the microbial assay. The new detection method was applied for rapid detection of bacteria (E.coli sp. and Lactobacillus sp.) and fungi (Mucor sp.), with limit of detection as low as single digit cells per mL within 10 min using a portable luminometer. We expect our simple and rapid detection method to be a powerful alternative to the conventional plate counting and immunoassay methods for rapid screening of microorganisms in food/drug products.

Keywords: microorganism testing, gold nanoplates, chemiluminescence, reducing agents, luminol

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3325 Fucoidan: A Potent Seaweed-Derived Polysaccharide with Immunomodulatory and Anti-inflammatory Properties

Authors: Tauseef Ahmad, Muhammad Ishaq, Mathew Eapen, Ahyoung Park, Sam Karpiniec, Vanni Caruso, Rajaraman Eri

Abstract:

Fucoidans are complex, fucose-rich sulfated polymers discovered in brown seaweeds. Fucoidans are popular around the world, particularly in the nutraceutical and pharmaceutical industries, due to their promising medicinal properties. Fucoidans have been shown to have a variety of biological activities, including anti-inflammatory effects. They are known to inhibit inflammatory processes through a variety of mechanisms, including enzyme inhibition and selectin blockade. Inflammation is a part of the complicated biological response of living systems to damaging stimuli, and it plays a role in the pathogenesis of a variety of disorders, including arthritis, inflammatory bowel disease, cancer, and allergies. In the current investigation, various fucoidan extracts from Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica were assessed for inhibition of pro-inflammatory cytokine production (TNF-α, IL-1β, and IL-6) in LPS induced human macrophage cell line (THP-1) and human peripheral blood mononuclear cells (PBMCs). Furthermore, we also sought to catalogue these extracts based on their anti-inflammatory effects in the current in-vitro cell model. Materials and Methods: To assess the cytotoxicity of fucoidan extracts, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5, -diphenyltetrazolium bromide) cell viability assay was performed. Furthermore, a dose-response for fucoidan extracts was performed in LPS induced THP-1 cells and PBMCs after pre-treatment for 24 hours, and levels of TNF-α, IL-1β, and IL-6 cytokines were measured using Enzyme-Linked Immunosorbent Assay (ELISA). Results: The MTT cell viability assay demonstrated that fucoidan extracts exhibited no evidence of cytotoxicity in THP-1 cells or PBMCs after 48 hours of incubation. The results of the sandwich ELISA revealed that all fucoidan extracts suppressed cytokine production in LPS-stimulated PBMCs and human THP-1 cells in a dose-dependent manner. Notably, at lower concentrations, the lower molecular fucoidan (5-30 kDa) extract from Macrocystis pyrifera was a highly efficient inhibitor of pro-inflammatory cytokines. Fucoidan extracts from all species including Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica exhibited significant anti-inflammatory effects. These findings on several fucoidan extracts provide insight into strategies for improving their efficacy against inflammation-related diseases. Conclusion: In the current research, we have successfully catalogued several fucoidan extracts based on their efficiency in LPS-induced macrophages and PBMCs in downregulating the key pro-inflammatory cytokines (TNF-, IL-1 and IL-6), which are prospective targets in human inflammatory illnesses. Further research would provide more information on the mechanism of action, allowing it to be tested for therapeutic purposes as an anti-inflammatory medication.

Keywords: fucoidan, PBMCs, THP-1, TNF-α, IL-1β, IL-6, inflammation

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3324 Molecular and Serological Diagnosis of Newcastle and Ornithobacterium rhinotracheale Broiler in Chicken in Fars Province, Iran

Authors: Mohammadjavad Mehrabanpour, Maryam Ranjbar Bushehri, Dorsa Mehrabanpour

Abstract:

Respiratory diseases are the most important problems in the country’s poultry industry, particularly when it comes to broiler flocks. Ornithobacterium rhinotracheale (ORT) is a species that causes poor performance in growth rate, egg production, and mortality. This pathogen causes a respiratory infection including pulmonary alveolar inflammation, and pneumonia of birds throughout the world. Newcastle disease (ND) is a highly contagious disease in poultry, and also, it causes considerable losses to the poultry industry. The aim of this study was to evaluate the simultaneous occurrence of ORT and ND and NDV isolation by inoculation in embryonated eggs and confirmed by RT-PCR in broiler chicken flocks in Fars province. In this study, 318 blood and 85 tissue samples (brain, trachea, liver, and cecal tonsils) were collected from 15 broiler chicken farms. Survey serum antibody titers against ORT by using a commercial enzyme-linked immunosorbent assay (ELISA) kit performed. Evaluation of antibody titer against ND virus is performed by hemagglutination inhibition test. Virus isolation with chick embryo eggs 9-11 and RT-PCR method were carried out. A total of 318 serum samples, 135 samples (42.5%) were positive for antibodies to ORT and titer of HI antibodies against NDV in 122 serum samples (38/4%) were 7-10 (log2) and 61 serum samples (19/2%) had occurrence antibody titer against Newcastle virus and ORT. Results of the present study indicated that 20 tissue samples were positive in embryonated egg and in rapid hemagglutination (HA) test. HI test with specific ND positive serum confirmed that 6 of 20 samples. PCR confirmed that all six samples were positive and PCR products of samples indicated 535-base pair fragments in electrophrosis. Due to the great economic importance of these two diseases in the poultry industry, it is necessary to design and implement a comprehensive plan for prevention and control of these diseases.

Keywords: ELISA, Ornithobacterium rhinotracheale, newcastle disease, seroprevalence

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3323 Methylprednisolone Injection Did Not Inhibit Anti-Hbs Response Following Hepatitis B Vaccination in Mice

Authors: P. O. Ughachukwu, P. O. Okonkwo, P. C. Unekwe, J. O. Ogamba

Abstract:

Background: The prevalence of hepatitis B viral infection is high worldwide with liver cirrhosis and hepatocellular carcinoma as important complications. Cases of poor antibody response to hepatitis B vaccination abound. Immunosuppression, especially from glucocorticoids, is often cited as a cause of poor antibody response and there are documented evidences of irrational administration of glucocorticoids to children and adults. The study was, therefore, designed to find out if administration of glucocorticoids affects immune response to vaccination against hepatitis B in mice. Methods: Mice of both sexes were randomly divided into 2 groups. Daily intramuscular methylprednisolone injections, (15 mg kg-1), were given to the test group while sterile deionized water (0.1ml) was given to control mice for 30 days. On day 6 all mice were given 2 μg (0.1ml) hepatitis B vaccine and a booster dose on day 27. On day 34, blood samples were collected and analyzed for anti-HBs titres using enzyme-linked immunosorbent assay (ELISA). Statistical analysis was done using Graph Pad Prism 5.0 and the results taken as statistically significant at p value < 0.05. Results: There were positive serum anti-HBs responses in all mice groups but the differences in titres were not statistically significant. Conclusions: At the dosages and length of exposure used in this study, methylprednisolone injection did not significantly inhibit anti-HBs response in mice following immunization against hepatitis B virus. By extrapolation, methylprednisolone, when used in the usual clinical doses and duration of therapy, is not likely to inhibit immune response to hepatitis B vaccinations in man.

Keywords: anti-HBs, hepatitis B vaccine, immune response, methylprednisolone, mice

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3322 Chikungunya Virus Infection among Patients with Febrile Illness Attending University of Maiduguri Teaching Hospital, Nigeria

Authors: Abdul-Dahiru El-Yuguda, Saka Saheed Baba, Tawa Monilade Adisa, Mustapha Bala Abubakar

Abstract:

Background: Chikungunya (CHIK) virus, a previously anecdotally described arbovirus, is now assuming a worldwide public health burden. The CHIK virus infection is characterized by potentially life threatening and debilitating arthritis in addition to the high fever, arthralgia, myalgia, headache and rash. Method: Three hundred and seventy (370) serum samples were collected from outpatients with febrile illness attending University of Maiduguri Teaching Hospital, Nigeria, and was used to detect for Chikungunya (CHIK) virus IgG and IgM antibodies using the Enzyme Linked Immunosorbent Assays (ELISAs). Result: Out of the 370 sera tested, 39 (10.5%) were positive for presence of CHIK virus antibodies. A total of 24 (6.5%) tested positive for CHIK virus IgM only while none (0.0%) was positive for presence of CHIK virus IgG only and 15 (4.1%) of the serum samples were positive for both IgG and IgM antibodies. A significant difference (p<0.0001) was observed in the distribution of CHIK virus antibodies in relation to gender. The males had prevalence of 8.5% IgM antibodies as against 4.6% observed in females. On the other hand 4.6% of the females were positive for concurrent CHIK virus IgG and IgM antibodies when compared to a prevalence of 3.4% observed in males. Only the age groups ≤ 60 years and the undisclosed age group were positive for presence of CHIK virus IgG and/or IgM antibodies. No significant difference (p>0.05) was observed in the seasonal prevalence of CHIK virus antibodies among the study subjects Analysis of the prevalence of CHIK virus antibodies in relation to clinical presentation (as observed by Clinicians) of the patients revealed that headache and fever were the most frequently encountered ailments. Conclusion: The CHIK virus IgM and concurrent IgM and IgG antibody prevalence rates of 6.5% and 4.1% observed in this study indicates a current infection and the lack of IgG antibody alone observed shows that the infection is not endemic but sporadic. Recommendation: Further studies should be carried to establish the seasonal prevalence of CHIK virus infection vis-à-vis vector dynamics in the study area. A comprehensive study need to be carried out on the molecular characterization of the CHIK virus circulating in Nigeria with a view to developing CHIK virus vaccine.

Keywords: Chikungunya virus, IgM and IgG antibodies, febrile patients, enzyme linked immunosorbent assay

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3321 Occurrence of Ranavirus in Edible Frogs and Fish Sold for Human Consumption in Kaduna State, Northern Nigeria

Authors: Inikpi Ameh, Grace Kia, A. K. B. Sackey, Joy Atawodi, Richard Whittington

Abstract:

Ranaviruses are belonging to the viral Family Iridoviridae, are a group of globally emerging pathogens recognized as major viral pathogens of cold-blooded vertebrates. They cause systemic infection in fishes, amphibians, and reptiles. Ranaviruses have been associated with numerous disease outbreaks in natural and cultured populations of fish, amphibians, and reptiles. To investigate the presence of the ranavirus in fish and edible frogs sourced from dams and ponds in Zaria, Kaduna State, Nigeria. A total of 425 frogs (Rana spp.) and fishes (n=215 and n=200, respectively) were randomly collected based on consent and availability. Liver, kidney, and spleen tissue samples from each animal were pooled and homogenized. The samples were screened for ranavirus using the Indirect Enzyme linked Immunosorbent assay (ELISA). An overall prevalence of 46.1% (196/425) was obtained from the study. Frogs had a prevalence of 51.2% (110/215) while fish had 43% (86/200). This is the first study on ranavirus in fish and edible frogs in Nigeria. This study has established that edible frogs (Rana spp) and fishes sold in Zaria, Nigeria were infected with ranavirus which may have great economic importance to the nation’s aquaculture. In view of occasional massive economic losses observed in fishery industry due to deaths of unknown origin, this preliminary investigation is useful in directing veterinarians, policy makers and researchers on need to survey for ranavirus and also enlighten the relevant stakeholders on its prevention and control in Nigeria.

Keywords: fish, frogs, Nigeria, Ranavirus

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3320 Origanum vulgare as a Possible Modulator of Testicular Endocrine Function in Mice

Authors: Eva Tvrdá, Barbora Babečková, Michal Ďuračka, Róbert Kirchner, Július Árvay

Abstract:

This study was designed to assess the in vitro effects of Origanum vulgare L. (oregano) extract on the testicular steroidogenesis. We focused on identifying major biomolecules present in the oregano extract, as well as to investigate its in vitro impact on the secretion of cholesterol, testosterone, dehydroepiandrosterone and androstenedione by murine testicular fragments. The extract was subjected to high performance liquid chromatography (HPLC) which identified cyranosid, daidzein, thymol, rosmarinic and trans-caffeic acid among the predominant biochemical components of oregano. For the in vitro experiments, testicular fragments from 20 sexually mature Institute of Cancer Research (ICR) mice were incubated in the absence (control group) or presence of the oregano extract at selected concentrations (10, 100 and 1000 μg/mL) for 24 h. Cholesterol levels were quantified using photometry and the hormones were assessed by ELISA (Enzyme-Linked Immunosorbent Assay). Our data revealed that the release of cholesterol and androstenedione (but not dehydroepiandrosterone and testosterone) by the testicular fragments was significantly impacted by the oregano extract in a dose-dependent fashion. Supplementation of the extract resulted in a significant decline of cholesterol (P < 0.05 in case of 100 μg/mL; P < 0.01 with respect 100 μg/mL extract), as well as androstenedione (P < 0.01 with respect to 100 and 1000 μg/mL extract). Our results suggest that the biomolecules present in Origanum vulgare L. could exhibit a dose-dependent impact on the secretion of male steroids, playing a role in the regulation of testicular steroidogenesis.

Keywords: mice, Origanum vulgare L., steroidogenesis, testes

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3319 Seroepidemiology of Q Fever among Companion Dogs in Fars Province, South of Iran

Authors: Atefeh Esmailnejad, Mohammad Abbaszadeh Hasiri

Abstract:

Coxiella burnetii is a gram-negative obligatory intracellular bacterium that causes Q fever, a significant zoonotic disease. Sheep, cattle, and goats are the most commonly reported reservoirs for the bacteria, but infected cats and dogs have also been implicated in the transmission of the disease to human. The aim of present study was to investigate the presence of antibodies against Coxiella burnetii among companion dogs in Fars province, South of Iran. A total of 181 blood samples were collected from asymptomatic dogs, mostly referred to Veterinary Hospital of Shiraz University for regular vaccination. The IgG antibody detection against Coxiella burnetii was made by indirect Enzyme-linked Immunosorbent Assay (ELISA), employing phase I and II Coxiella burnetii antigens. A logistic regression model was developed to analyze multiple risk factors associated with seropositivity. An overall seropositivity of 7.7% (n=14) was observed. Prevalence was significantly higher in adult dogs above five years (18.18 %) compared with dogs between 1 and five years (7.86 %) and less than one year (6.17%) (P=0.043). Prevalence was also higher in male dogs (11.21 %) than in female (2.7 %) (P=0.035). There were no significant differences in the prevalence of positive cases and breed, type of housing, type of food and exposure to other farm animals (P>0.05). The results of this study showed the presence of Coxiella burnetii infection among the companion dogs population in Fars province. To our knowledge, this is the first study regarding Q fever in dogs carried out in Iran. In areas like Iran, where human cases of Q fever are not common or remain unreported, the public health implications of Q fever seroprevalence in dogs are quite significant.

Keywords: Coxiella burnetii, dog, Iran, Q fever

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3318 Profiling of Apoptotic Protein Expressions after Trabectedin Treatment in Human Prostate Cancer Cell Line PC-3 by Protein Array Technology

Authors: Harika Atmaca, Emir Bozkurt, Latife Merve Oktay, Selim Uzunoglu, Ruchan Uslu, Burçak Karaca

Abstract:

Microarrays have been developed for highly parallel enzyme-linked immunosorbent assay (ELISA) applications. The most common protein arrays are produced by using multiple monoclonal antibodies, since they are robust molecules which can be easily handled and immobilized by standard procedures without loss of activity. Protein expression profiling with protein array technology allows simultaneous analysis of the protein expression pattern of a large number of proteins. Trabectedin, a tetrahydroisoquinoline alkaloid derived from a Caribbean tunicate, Ecteinascidia turbinata, has been shown to have antitumor effects. Here, we used a novel proteomic approach to explore the mechanism of action of trabectedin in prostate cancer cell line PC-3 by apoptosis antibody microarray. XTT cell proliferation kit and Cell Death Detection Elisa Plus Kit (Roche) was used for measuring cytotoxicity and apoptosis. Human Apoptosis Protein Array (R&D Systems) which consists of 35 apoptosis related proteins was used to assess the omic protein expression pattern. Trabectedin induced cytotoxicity and apoptosis in prostate cancer cells in a time and concentration-dependent manner. The expression levels of the death receptor pathway molecules, TRAIL-R1/DR4, TRAIL R2/DR5, TNF R1/TNFRSF1A, FADD were significantly increased by 4.0-, 21.0-, 4.20- and 11.5-fold by trabectedin treatment in PC-3 cells. Moreover, mitochondrial pathway related pro-apoptotic proteins Bax, Bad, Cytochrome c, and Cleaved Caspase-3 expressions were induced by 2.68-, 2.07-, 2.8-, and 4.5-fold and the expression levels of anti-apoptotic proteins Bcl-2 and Bcl-XL were reduced by 3.5- and 5.2-fold in PC-3 cells. Proteomic (antibody microarray) analysis suggests that the mechanism of action of trabectedin may be exerted via the induction of both intrinsic and extrinsic apoptotic pathways. The antibody microarray platform can be utilised to explore the molecular mechanism of action of novel anticancer agents.

Keywords: trabectedin, prostate cancer, omic protein expression profile, apoptosis

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3317 Host Responses in Peri-Implant Tissue in Comparison to Periodontal Tissue

Authors: Raviporn Madarasmi, Anjalee Vacharaksa, Pravej Serichetaphongse

Abstract:

The host response in peri-implant tissue may differ from that in periodontal tissue in a healthy individual. The purpose of this study is to investigate the expression of inflammatory cytokines in peri-implant crevicular fluid (PICF) from single implant with different abutment types in comparison to healthy periodontal tissue. 19 participants with healthy implants and teeth were recruited according to inclusion and exclusion criteria. PICF and gingival crevicular fluid (GCF) was collected using sterile paper points. The expression level of inflammatory cytokines including IL-1α, IL-1β, TNF-α, IFN-γ, IL-6, and IL-8 was assessed using enzyme-linked immunosorbent assay (ELISA). Paired t test was used to compare the expression levels of inflammatory cytokines around natural teeth and peri-implant in PICF and GCF of the same individual. The Independent t-test was used to compare the expression levels of inflammatory cytokines in PICF from titanium and UCLA abutment. Expression of IL-6, TNF-α, and IFN-γ in PICF was not statistically different from GCF among titanium and UCLA abutment group. However, the level of IL-1α in the PICF from the implants with UCLA abutment was significantly higher than GCF (P=0.030). In addition, the level of IL-1β in PICF from the implants with titanium abutment was significantly higher than GCF (P=0.032). When different abutment types was compared, IL-8 expression in PICF from implants with UCLA abutment was significantly higher than titanium abutment (P=0.003).

Keywords: abutment, dental implant, gingival crevicular fluid and peri-implant crevicular fluid

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3316 HLB Disease Detection in Omani Lime Trees using Hyperspectral Imaging Based Techniques

Authors: Jacintha Menezes, Ramalingam Dharmalingam, Palaiahnakote Shivakumara

Abstract:

In the recent years, Omani acid lime cultivation and production has been affected by Citrus greening or Huanglongbing (HLB) disease. HLB disease is one of the most destructive diseases for citrus, with no remedies or countermeasures to stop the disease. Currently used Polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) HLB detection tests require lengthy and labor-intensive laboratory procedures. Furthermore, the equipment and staff needed to carry out the laboratory procedures are frequently specialized hence making them a less optimal solution for the detection of the disease. The current research uses hyperspectral imaging technology for automatic detection of citrus trees with HLB disease. Omani citrus tree leaf images were captured through portable Specim IQ hyperspectral camera. The research considered healthy, nutrition deficient, and HLB infected leaf samples based on the Polymerase chain reaction (PCR) test. The highresolution image samples were sliced to into sub cubes. The sub cubes were further processed to obtain RGB images with spatial features. Similarly, RGB spectral slices were obtained through a moving window on the wavelength. The resized spectral-Spatial RGB images were given to Convolution Neural Networks for deep features extraction. The current research was able to classify a given sample to the appropriate class with 92.86% accuracy indicating the effectiveness of the proposed techniques. The significant bands with a difference in three types of leaves are found to be 560nm, 678nm, 726 nm and 750nm.

Keywords: huanglongbing (HLB), hyperspectral imaging (HSI), · omani citrus, CNN

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3315 Current Epizootic Situation of Q Fever in Polish Cattle

Authors: Monika Szymańska-Czerwińska, Agnieszka Jodełko, Krzysztof Niemczuk

Abstract:

Q fever (coxiellosis) is an infectious disease of animals and humans causes by C. burnetii and widely distributed throughout the world. Cattle and small ruminants are commonly known as shedders of C. burnetii. The aims of this study were the evaluation of seroprevalence and shedding of C. burnetii in cattle. Genotypes of the pathogen present in the tested specimens were also identified using MLVA (Multiple Locus Variable-Number Tandem Repeat Analysis) and MST (multispacer sequence typing) methods. Sampling was conducted in different regions of Poland in 2018-2021. In total, 2180 bovine serum samples from 801 cattle herds were tested by ELISA (enzyme-linked immunosorbent assay). 489 specimens from 157 cattle herds such as: individual milk samples (n=407), bulk tank milk (n=58), vaginal swabs (n=20), placenta (n=3) and feces (n=1) were subjected to C. burnetii specific qPCR. The qPCR (IS1111 transposon-like repetitive region) was performed using Adiavet COX RealTime PCR kit. Genotypic characterization of the strains was conducted utilizing MLVA and MST methods. MLVA was performed using 6 variable loci. The overall herd-level seroprevalence of C. burnetii infection was 36.74% (801/2180). Shedders were detected in 29.3% (46/157) cattle herds in all tested regions. ST 61 sequence type was identified in 10 out of 18 genotyped strains. Interestingly one strain represents sequence type which has never been recorded previously. MLVA method identified three previously known genotypes: most common was J but also I and BE were recognized. Moreover, a one genotype has never been described previously. Seroprevalence and shedding of C. burnetii in cattle is common and strains are genetically diverse.

Keywords: Coxiella burnetii, cattle, MST, MLVA, Q fever

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3314 Soluble CD36 and Cardiovascular Risk in Middle-Aged Subjects

Authors: Mohammad Alkhatatbeh, Nehad Ayoub, Nizar Mhaidat, Nesreen Saadeh, Lisa Lincz

Abstract:

CD36 is involved in the development of atherosclerosis by enhancing macrophage endocytosis of oxidized-low density lipoproteins and foam cell formation. Soluble CD36 (sCD36) was found to be elevated in type 2 diabetic patients and was supposed to act as a marker of insulin resistance and atherosclerosis. In young subjects, sCD36 was associated with cardiovascular risk factors including obesity and hypertriglyceridemia. This study was conducted to further investigate the relationship between plasma sCD36 and cardiovascular risk factors among middle-aged patients with metabolic syndrome (MetS) and healthy controls. SCD36 concentrations were determined by enzyme-linked immunosorbent assays (ELISA) for 41 patients with MetS and 36 healthy controls. Data for other variables were obtained from patients' medical records. SCD36 concentrations were relatively low compared to most other studies and were not significantly different between the MetS group and controls (P-value=0.17). SCD36 was also not correlated with age, body mass index, glucose, lipid profile, serum electrolytes and blood counts. SCD36 was not significantly different between subjects with obesity, hyperglycemia, dyslipidemia, hypertension or cardiovascular disease and those without these abnormalities (P-value > 0.05). The inconsistency between results reported in this study and other studies may be unique to the study population or be a result of the lack of a reliable standardized method for determining absolute sCD36 concentrations. However, further investigations are required to assess CD36 tissue expression in the study population and to assess the accuracy of various commercially available sCD36 ELISA kits. Thus, the availability of a standardized simple sCD36 ELISA that could be performed in any basic laboratory would be more favorable to the specialized flow cytometry methods that detect CD36+ microparticles if it was to be used as a biomarker.

Keywords: metabolic syndrome, CD36, cardiovascular risk, obesity, type 2 diabetes mellitus

Procedia PDF Downloads 245
3313 Role of Pro-Inflammatory and Regulatory Cytokines in Pathogenesis of Graves’ Disease in Association with Autoantibody Thyroid and Regulatory FoxP3 T-Cells

Authors: Dwitya Elvira, Eryati Darwin

Abstract:

Background: Graves’ disease (GD) is an autoimmune thyroid disease. Imbalance of Th1/Th2 cells and T-regulatory (Treg)/Th17 cells was thought to play pivotal role in the pathogenesis of GD. Treg FoxP3 produced TGF-β to maintain regulatory function, and Th17 cells produced IL-17 as cytokines that were thought in mediating several autoimmune diseases. The aim of this study is to assess the role of IL-17 and TGF-β in the pathogenesis of GD and to investigate its correlation with Thyroid Stimulating Hormone Receptor Antibody (TRAb) and Treg FoxP3 expression. Method: 30 GD patients and 27 age and sex-matched controls were enrolled in this study. Diagnosis of GD was based on clinical and biochemical of GD. Serum IL-17, TGF-β, TRAb, and FoxP3 were measured by enzyme-linked immunosorbent assay (ELISA). Data were analyzed by using SPSS 21.0 (SPSS Inc.). Spearman rank correlation test was used for assessment of correlation. The statistical significance was accepted as P<0.05. Result: There was no significant correlation between IL-17 and TGF-β serum with expression of FoxP3 level in GD, but there was significant correlation between TGF-β and TRAb serum level (P<0.05). Serum levels of IL-17 and TGF-β were found to be elevated in patient group compared to control, where mean values of IL-17 were 14.43±2.15 pg/mL and TGF-β were 10.44±3.19 pg/mL in patients group; and in control group, level of IL-17 were 7.1±1.45 pg/mL and TGF-β were 4.95±1.35 pg/mL. Conclusion: Serum Il-17 and TGF-β were elevated in GD patients that reflect the role of inflammatory and regulatory cytokines activation in pathogenesis of GD. There was significant correlation between TGF-β and TRAb, revealing that Treg cytokines may play a role in pathogenesis of GD.

Keywords: IL-17, TGF-B, FoxP3, TRAb, Graves’ disease

Procedia PDF Downloads 249
3312 Survey of Potato Viral Infection Using Das-Elisa Method in Georgia

Authors: Maia Kukhaleishvili, Ekaterine Bulauri, Iveta Megrelishvili, Tamar Shamatava, Tamar Chipashvili

Abstract:

Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus strain. Selection of resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according potato cultivars for further selection of virus-free material in Georgia. During the summer 2015- 2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), Potato virus M (PVM), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY) and potato leaf roll virus (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DASELISA) was used at the laboratory to analyze the results. The result showed that PVY (21.4%) and PLRV (19.7%) virus presence in collected samples was relatively high compared to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentrations. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia) and PVM virus (3.12%) was characterized with low prevalence. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9% of samples were containing PVY/PLRV mix infection. Based on the results it can be concluded that PVY and PLRV infections are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term control of potato viral infection, especially seed-potatoes, must be regarded as the most important factor to increase seed productivity.

Keywords: virus, potato, infection, diseases

Procedia PDF Downloads 258
3311 Seroprevalence and Associated Factors of Hepatitis B and Hepatitis C Viral Infections among Prisoners in Tigrai, Northern Ethiopia

Authors: Belaynesh Tsegay Beyene, Teklay Gebrecherkos, Atsebaha Gebrekidan Kahsay, Mahmud Abdulkader

Abstract:

Background: Hepatitis B and C viruses are of important health and socioeconomic problem of the globe with remarkable diseases and deaths in Sub-Saharan African countries. The burden of hepatitis is unknown in the prison settings of Tigrai. Therefore, we aimed to describe the seroprevalence and associated factors of hepatitis B and C viruses among prisoners of Tigrai, Ethiopia. Methods: A cross-sectional study was carried out from February 2020 to May 2020 at the prison facilities of Tigrai. Demographics and associated factors were collected from 315 prisoners prospectively. Five milliliter of blood was collected and tested using rapid tests kits of HBsAg (Zhejiang orient Gene Biotech Co., Ltd., China) and HCV antibodies (Volkan Kozmetik Sanayi Ve Ticaret Ltd. STI, Turkey). Positive samples were confirmed using enzyme-linked immunosorbent assay (ELISA) (Beijing Wantai Biological Pharmacy Enterprise Co. Ltd). Data were analyzed using Statistical Package for Social Sciences (SPSS) version 20 and p < 0.05 was considered statistically significant. Results: The overall seroprevalence of HBV and HCV were 25 (7.9%) and 1(0.3%), respectively. The majority of hepatitis B viral infections were identified from the age groups of 18-25 years (10.7%) and unmarried prisoners (11.8%). Prisoners greater than 100 per cell [AOR =3.95, 95% CI= (1.15, 13.6, p =0.029)] and having history of alcohol consumption [AOR =3.01, 95% CI= (1.17, 7.74, p =0.022)] were significantly associated with HBV infections. Conclusions: The seroprevalence of HBV among prisoners was nearly high or borderline (7.9%) with a very low HCV prevalence (0.3%). HBV was most prevalent among young adults, large number of prisoners per cell and those who had history of alcohol consumption. This study recommends that there should be prison-focused intervention including regular health education by emphasis on the mode of transmission and introducing HBV screening policy for prisoners especially when they enter to the prison.

Keywords: seroprevalence, HBV, HCV, prisoners, Tigrai

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3310 Comparison of Transforming Growth Factor-β1 Levels in the Human Gingival Sulcus during Canine Retraction Using Elastic Chain and Closed Coil Spring

Authors: Sri Suparwitri

Abstract:

When an orthodontic force is applied to a tooth, an inflammatory response is initiated then lead to bone remodeling process, and the process accommodates tooth movement. One of cytokine that plays a prominent role in bone remodeling process was transforming growth factor-beta 1 (TGF-β1). The purpose of this study was to identify and compare changes of TGF-β1 in human gingival crevicular fluid during canine retraction using elastic chain and closed coil spring. Ten patients (mean age 20.7 ± 2.9 years) participated. The patients were entering the space closure phase of fixed orthodontic treatment. An upper canine of each patient was retracted using elastic chain, and the contralateral canine was retracted using closed coil spring. Gingival crevicular fluid samples were collected from the canine teeth before and 7 days after the force was applied. Transforming growth factor-beta 1 was determined by enzyme-linked immunosorbent assay (ELISA). The concentrations of TGF-β1 at 7 days were significantly higher compared to before canine retraction in both groups. In the evaluation of between-group difference, before retraction, the difference was insignificant, whereas at 7 days significantly higher values were determined in the closed coil spring group compared to elastic chain group. The result suggests that TGF-β1 is associated with the bone remodeling that occurs during canine distalization movement. Closed coil spring gave higher TGF-β1 concentrations thus more bone remodeling occurred and may be considered the treatment of choice.

Keywords: closed coil spring, elastic chain, gingival crevicular fluid, TGF-β1

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3309 The Effect of Environmental Enrichment on Anxiety and Stress Hormone in Maternally Separated Male Rats

Authors: Özge Selin Çevik, Leyla Şahin, Gülhan Örekeci Temel

Abstract:

The early postnatal period is critical for the development of cognitive and emotional functions. Maternal separation is a detrimental postnatal influence, whereas environmental enrichment is a therapeutic and protective agent. It is unclear if long-term environmental enrichment can compensate for the effects of maternal separation stress on anxiety behavior. This study was designed to examine how environmental enrichment affects anxiety levels and corticosterone levels in maternally separated rats. There are six main groups in this study: control (C), maternal separation+standard cage (MS), maternal separation+enriched environment (MSE), enriched environment (E), the maternal separation that decapitated at postnatal (PN) 21 (MS21), and standard cage that decapitated at PN21 (STD21). The maternal separation procedure consisted of PN for 21 days (between 09:00 a.m and 12:00 a.m). Enriched (E, MSE) or standard cage environment rats (MS, C) spent PN (22-55) days in either enriched cages or standard cages. Anxiety and locomotor activity were examined with the open field and elevated plus-maze test. Blood corticosterone level was evaluated by the enzyme-linked immunosorbent assay (ELISA) method. Results showed that maternal separation (MS) increased locomotor activity and anxiety. An enriched environment (E) did not change the locomotor activity. MSE group’s anxiety and locomotor activity did not change. Corticosterone levels increased in the maternal separation group that decapitated at the PN 21 days. Maternal separation increases anxiety. Environmental enrichment alone was insufficient to cause alterations in the anxiety level. In addition, environmental enrichment did not ameliorate the anxiety level in maternally separated rats. However, environmental enrichment decreased the locomotor activity in the maternally separated rats.

Keywords: maternal separation, environment enrichment, stress, hippocampus, anxiety, memory, rat

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3308 Immobilization Strategy of Recombinant Xylanase from Trichoderma reesei by Cross-Linked Enzyme Aggregates

Authors: S. Md. Shaarani, J. Md. Jahim, R. A. Rahman, R. Md. Illias

Abstract:

Modern developments in biotechnology have paved the way for extensive use of biocatalysis in industries. Although it offers immense potential, industrial application is usually hampered by lack of operational stability, difficulty in recovery as well as limited re-use of the enzyme. These drawbacks, however, can be overcome by immobilization. Cross-linked enzyme aggregates (CLEAs), a versatile carrier-free immobilization technique is one that is currently capturing global interest. This approach involves precipitating soluble enzyme with an appropriate precipitant and subsequent crosslinking by a crosslinking reagent. Without ineffective carriers, CLEAs offer high enzymatic activity, stability and reduced production cost. This study demonstrated successful CLEA synthesis of recombinant xylanase from Trichoderma reesei using ethanol as aggregating agent and glutaraldehyde (2% (v/v); 100 mM) as crosslinker. Effects of additives including proteic feeder such as bovine serum albumin (BSA) and poly-L-Lysine were investigated to reveal its significance in enhancing the performance of enzyme. Addition of 0.1 mg BSA/U xylanase showed considerable increment in CLEA development with approximately 50% retained activity.

Keywords: cross-linked, immobilization, recombinant, xylanase

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3307 Levels of Microcystin in the Coastal Waters of Nigeria

Authors: Medina Kadiri

Abstract:

Blue-green otherwise called cyanobacteria, produce an array of biotoxins grouped into five categories notably hapatotoxins, neurotoxins, cytotoxins, dermatotoxins, and irritant toxins. Microcystins which are examples of hepatotoxins produced by blue-green algae Microcystins comprise the most common group of the cyanobacterial toxins. Blue-green algae flourish in aquatic environments, whether marine, brackish or freshwater, producing blooms in different forms such as microscopic, mats, or unsightly odoriferous scums. Microcystins biotoxins cause a plethora of animal and human hazards such as liver damage/cirrhosis and cancer, kidney damage, dermatitis, tinnitus, gastroenteritis, sore throat, nausea, myalgia, neurological problems, respiratory irritation and death. Water samples were collected from coastal regions of Nigeria in March 2014, June 2014, October 2014 and January 2015 and analyzed with Enzyme Linked Immunosorbent Assay (ELISA) kits. Microcystin biotoxin was recorded in all sites both during dry and wet seasons. The range of microcystins found was 0.000041-There was a seasonal trend of increasing microcystin concentrations from March till Octobers and a decrease thereafter. Generally in the oceanic waters, microcystin levels were highest at Cross Rivers in March and January, Barbeach in June and Lekki in October. In the adjoining riverine ecosystems, on the other hand, the highest concentrations of microcystin were observed at Akwa Ibom in March, June and October and in Bayelsa in January. Continuous monitoring and screening of coastal water bodies is suggested to minimize the health risks of cyanobacterial biotoxins to coastal communities of Nigeria.

Keywords: biotoxins, harmful algae, marine, microcystin, Nigeria

Procedia PDF Downloads 258
3306 The Role of Neuroserpin in Rheumatoid Arthritis Patients

Authors: Sevil Arabaci Tamer, Gonul Gurol, Ibrahim Tekeoglu, Halil Harman, Ihsan Hakki Ciftci

Abstract:

Neuroserpin (NSP) is a serine protease inhibitor and member of the serpin family. It is expressed in developing and adult nervous systems, and acts as an inhibitor of protease tissue plasminogen activator (tPA) and a regulator of neuronal growth and plasticity. Also NSP displays anti-inflammatory activity. But, its role in rheumatoid arthritis had never been studied before. So, the aim of the present study was to investigate the effect of neuroserpin in patients with RA. A total of 50 frozen (-20 ºC) serum samples 40 of them belonged to patients with RA, and 10 sample belonged to healthy subjects, were enrolled prospectively. We used DAS-28 to evaluate disease activity. The following clinical data gathered from the original patients' charts. Serum neuroserpin levels were measured by enzyme-linked immunosorbent assay. Our preliminary study results demonstrate, for the first time, that NSP levels are significantly different in RA patients relative to healthy subjects (P = 0.014). So, NSP contribute to pathological condition of RA. Thus, we believe that serum NSP levels can be as a marker in patients with RA. However other inflammatory diseases should be further investigated.

Keywords: neuroserpin, rheumatoid arthritis, tPA, tPA inhibitor

Procedia PDF Downloads 445