Search results for: In Vitro isolated microspore culture

Authors: A. Attahiru, Rabi Abdullahi Danjuma, Fatima Bint

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Language and culture are two concepts which are closely related that one affects the other. This paper attempts to examine the definition of language and culture by discussing the relationship between them. The paper further presents some instructional strategies for the teaching of language and culture as well as the influence of culture on language. It also looks at its implication to language education and finally some recommendation and conclusion were drawn.

Keywords: culture, language, relationship, strategies, teaching

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Authors: Isaac Samuel

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A study was conducted to determine the in-vitro degradation of selected Nigerian browse plants consumed by small ruminants on free range in northern guinea savannah region of Nigeria using in vitro gas production, proximate composition, fibre components, methane gas production and dry matter degradation as tools. The leaves samples of the selected browse plants were collected, processed and incubated using in vitro gas dry matter degradation techniques. Results obtained showed variation in the rate of degradation. The result obtained from chemical analysis showed that the CP content of A. occidentale (26.49%) was higher than F. thonningi (23.58%), M. indica (20.58%) and T. catappa (18.61%). Both ADF and NDF of A. occidentale (40.00 and 50.00) were as well higher than F. thonningi (20.00 and 40.00), M. indica (20.00 and 40.00) and T.catappa (20.00 and 42.00). Results from in vitro gas production however showed that T. catappa (23.67ml/DM) has a significantly higher (p<0.05) value than F.thonningi (20.67ml/DM), A. occidentale (16.67ml/DM), and M. indica(14.00ml/DM) at 72 hours of incubation. Methane gas production and in vitro gas production can be used to predict dry matter degradation and nutritive value of feedstuff for small ruminants. A. occidentale with the least methane gas production and highest crude protein (CP) content might have the most nutritive value among the browse plants investigated.

Keywords: in vitro, degradation, browse, gas production

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Authors: S. I. Nwadioha, M. S. Odimayo, J. A. Omotayo, A. Olu Taiwo, O. E. Olabiyi

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The current study was conducted to determine the epidemiology and antibiotic sensitivity pattern of bacteria isolated from blood of septicemic patients for improved antibiotic therapy. A three-year descriptive study has been carried out at Microbiology Laboratory, Ekiti State University Teaching Hospital, Ado Ekiti, from April 2012 to April 2015. Information compiled from patients’ records includes age, sex, isolated organisms and antibiotic susceptibility patterns. Three hundred and thirteen blood cultures were collected from neonatology and pediatrics wards, Out Patients’ Department (OPD) and from other adult patients. Forty-one cultures yielded mono microbial growth (no polymicrobial growth), giving an incidence of 13.1% positive blood culture (N=41/313). There were 58.4% Gram-negative bacilli and 41.6% Gram-positive cocci in the microbial growth. Bacteria isolated were Staphylococcus aureus 34%(14/41), Klebsiella species22% (9/41), Enterococci 17%(7/41), Proteus species12%(5/41), Escherichia coli 7%(3/41) and Streptococcal pneumoniae 7%(3/41). There was a (35%) higher occurrence of septicemia in neonates than in any other age groups in the hospital. Bacterial sensitivity to 13 antibiotic agents was determined by antibiotics disc diffusion using modified Kirby Bauer’s method. Gram-positive organisms showed a higher antibiotic sensitivity ranging from 14- 100% than the Gram-negative bacteria (11-80%). Staphylococcus aureus and Klebsiella species are the most prevalent organisms. The third generation Cephalosporins (Ceftriaxone) and Floroquinolone(Levofloxacin, Ofloxacin) have proved reliable for management of these blood infections.

Keywords: blood cultures, septicemia, antibiogram, Nigeria

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Authors: Sihem Benaouda

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English is gaining an international identity as it affects every academic and professional field in the world. Without increasing their cultural understanding, it would obviously be difficult to completely educate learners for communication in a globalised environment. The concept of culture is intricate and needs to be elucidated, especially in an English language teaching (ELT) context. The study focuses on the investigation of the cultural studies integrated into the different types of English for specific purposes (ESP) materials, as opposed to English for general purposes (EGP) textbooks. A qualitative methodology based on a triangulation of techniques was conducted through materials analysis of five textbooks in both advanced EGP and three types of ESP. In addition to a semi-structured interview conducted with Algerian ESP practitioners, data analysis results revealed that culture in ESP textbooks is not overtly isolated into chapters and that cultural studies are predominantly present in business and economics materials, namely English for hotel and catering staff, tourism, and flight attendants. However, implicit cultural instruction is signalled in the social sciences and is negligible in science and technology sources. In terms of content, cultural studies in EGP are more related to generic topics, whereas, in some ESP materials, the topics are rather oriented to the specific field they belong to. Furthermore, the respondents’ answers showed an unawareness of the importance of culture in ESP teaching, besides some disregard for culture teaching per se in ESP contexts.

Keywords: ESP, EGP, cultural studies, textbooks, teaching, materials

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Authors: Kara Terki Ibtissem, Hassaine Hafida, Bellifa Samia

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Introduction: Staphylococcus aureus is one of the species that are most frequently isolated from urinary catheters. The ability to produce a biofilm is an important step in the pathogenesis of these staphylococci; biofilm formation is strongly dependent on the environmental conditions it also depends on the different parameters these biofilms are subjected to. Antiseptics, including ethanol and betadine, are used in clinical practice for disinfection and infection prevention. Recent studies, however, demonstrate that disinfectants may enhance biofilm production in Staphylococci. Methods: In this study, 48 staphylococcus aureus isolated from urinary catheters at the University Hospital Center of Sidi Bel Abbes (in Northwestern Algeria) were analyzed to detect the formation of biofilm by culture on Red Congo Agar (RCA), the Tube Method (TM) and tissue Culture Plate (TCP) techniques, this last was also used to investigate the effect of ethanol and Betadine on the preformed biofilm In a second time to know which environment is most favorable to the formation of the biofilm we perform a statistical test based on the student test by the software R. Results: It has been found that 23 strains produced a bacterial slime on the Congo red medium, 5 strains produced a biofilm by the tube method, 2 of which are highly productive. In addition, 7 strains produced a biofilm on polystyrene micro-plates; this number was higher in the presence of ethanol 70% and ethanol 90% with 19 and 11 biofilm-producing strains, respectively. On the other hand, no biofilm was formed in the presence of Betadine. Conclusion: It is important to examine the response of biofilms following an imposed external constraint, such as disinfectants, in order to develop new strategies to combat bacterial biofilms but also to better control their formation.

Keywords: staphylococcus aureus, biofilm, urinary catheter, ethanol

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Authors: Akwu A. Nneka, Naidoo, Yougasphree

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A significantly high number of microbes exist in higher plants; these microbes include bacteria, fungi, and actinomycetes. There are reports on the benefits of endophytic fungi and their products of metabolism to the host plant and man, consequently, it is expedient to explore the changes that could arise as a result of manipulating their growth media. Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is an indigenous Southern African plant, that belongs to a genus with known medicinal properties. Three media were used to culture the endophytic fungi viz., Potato Dextrose Agar (PDA), Malt Extract Agar (MEA), and Bacteriological Agar (BA) were used singly, and supplemented with three dilutions of the leaves and stem bark extracts. The manipulated growth media composition had a significant effect on the diversity of the isolated fungal populations. Several endophytic fungi were isolated; their distribution and diversity revealed a significant relatedness with the manipulated media. The media supplemented with the plant extracts was observed to give a significant increase in the growth rate and yield of the endophytes. To the best of our knowledge, this is the first study describing the endophytic fungi present in the leaves and stem bark of G. lasiocarpa E. Mey. ex Harv.

Keywords: Grewia lasiocarpa, plant-based extracts, endophytic fungi, Malvaceae

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Authors: Emmanuel O Ajani, Sabiu S, Mariam Zakari, Fisayo A Bamisaye

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Hydrocotyl bonariensis is a plant which anticataractogenic potentials have been reported. In the present study an attempt was made to evaluate the in vitro antioxidant activity of the fractionates of the leaves extract and also characterize some of its chemical constituents. DPPH, H₂O₂, OH and NO free radical scavenging, metal chelating and reducing power activity was used to evaluate the antioxidant activity of the crude extract fractionates. Fresh leaves of Hydrocotyl bonariensis leaves were extracted in 70% methanol. The extract was partitioned with different solvent system of increasing polarity (n-hexane, chloroform, ethyl acetate methanol and water). Compounds were isolated from the aqueous practitionate using accelerated gradient chromatography, vacuum liquid chromatography, preparative TLC and conventional column chromatography. The presence of the chemical groups was established with HPLC and Fourier Transform Infra Red. The structures of isolated compounds were elucidated by spectroscopic study and chemical shifts. Data from the study indicates that all the fractionates contain compounds with free radical scavenging activity. This activity was more pronounced in the aqueous fractionate (DPPH IC₅₀, 0025 ± 0.011 mg/ml, metal chelating capacity 27.5%, OH- scavenging IC₅₀, 0.846 ± 0.037 mg/ml, H₂O₂ scavenging IC₅₀ 0.521 ± 0.015 mg/ml, reducing power IC₅₀ 0.248 ± 0.025 mg/ml and NO scavenging IC₅₀ 0.537 ± 0.038 mg/ml). Two compounds were isolated and when compared with data from the literature; the structures were suggestive of polyphenolic flavonoid, quercetin and 3-O-β-D-glucopyranosyl-sitosterol. The result indicates that H. bonariensis leaves contain bioactive compounds with antioxidant activity.

Keywords: antioxidant, cataract, free radical, flavonoids, hydrocotyl bonariensis

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Authors: Philip Vercoe, Ali Hardan

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Samples of 29 Australian wild natives and exotic plants were tested in vitro batch rumen culture system for their methanogenic characteristics and potential usage as feed or antimicrobial to enhance sustainable livestock ruminant production system. The plants were tested for their in vitro rumen fermentation end products properties which include: methane production, total gas pressure, concentrations of total volatile fatty acids, ammonia, and acetate to propionate ratio. All of the plants were produced less methane than the positive control (i.e., oaten chaff) in vitro. Nearly 50 % of plants inhibiting methane by over 50% in comparison to the control. Eremophila granitica had the strongest inhibitory effect about 92 % on methane production comparing with oaten chaff. The exotic weed Arctotheca calendula (Capeweed) had the highest concentration of volatile fatty acids production as well as the highest in total gas pressure among all plants and the control. Some of the acacia species have the lowest production of total gas pressure. The majority of the plants produced more ammonia than the oaten chaff control. The plant species that produced the most ammonia was Codonocarpus cotinifolius, producing over 3 times as much methane as oaten chaff control while the lowest was Eremophila galeata. There was strong positive correlation between methane production and total gas production as well as between total gas production and the concentration of VFA produced with R² = 0.74, R² = 0.84, respectively. While there was weak positive correlation between methane production and the acetate to propionate ratio as well as between the concentration of VFA produced and methane production with R² = 0.41, R² = 0.52, respectively.

Keywords: in vitro Rumen Fermentation, methane, wild Australian native plants, forages

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Authors: N. Selami, M. Kaid Harche

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A pink-pigmented, aerobic, facultatively methylotrophic bacterium, was isolated from Retama monosperma root nodules and identified as a member of the genus Methylobacterium. Inoculation of R. monosperma plants by a pure culture of isolate strain under a hydroponic condition, resulted, 10 dpi, the puffiness at lateral roots. The observation in detail the anatomy and ultra-structure of infection sites by light and electron microscopy show that the bacteria induce stimulation of the division of cortical cells and digestion of epidermis cells then, Methylobacterium was observed in the inter and intracellular spaces of the outer cortex root. These preliminary results allow us to suggest the establishment of an epi-endosymbiotic interaction between Methylobacterium sp and R. monosperma.

Keywords: endophytic colonization, Methylobacterium, microscopy, nodule, pink pigmented, Retama monosperma

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Authors: Elyes Zarrouk, Yassine Benayed, Faiez Gargouri

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This paper presents a new hybrid model for isolated Arabic words recognition. To do this, we apply Support Vectors Machine (SVM) as an estimator of posterior probabilities within the Dynamic Bayesian networks (DBN). This paper deals a comparative study between DBN and SVM/DBN systems for multi-dialect isolated Arabic words. Performance using SVM/DBN is found to exceed that of DBNs trained on an identical task, giving higher recognition accuracy for four different Arabic dialects. In fact, the average of recognition rates for the four dialects with SVM/DBN was 87.67% while 83.01% with DBN.

Keywords: dynamic Bayesian networks, hybrid models, supports vectors machine, Arabic isolated words

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Authors: Dalvir Kataria, Khushminder Kaur Chahal, Amit Kumar

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The carrot seed essential oil was obtained by hydrodistillation. Hexane, dichloromethane, and methanol solvents were used for extraction of carrot seed by Soxhlet extraction methods. The major and minor compounds identified in carrot seed essential oil were carotol (52.73), daucol (5.10), daucene (5.68), (E)-β-farnesene (5.40), β-cubebene (3.19), longifolenaldehyde (3.23), β-elimene (3.23), (E)-caryophyllene (1.22), β-bisabolene (2.95) etc. The chemical composition of hexane, dichloromethane, and methanol extracts was different. Carotol was the common compound present. Major compounds isolated were from the carrot seed essential oil by column chromatography. Chemical transformations of carotol (2) with mercuric acetate/sodium borohydride, dry hydrochloric acid gas, acetonitrile/sulfuric acid, selenium dioxide/t-butyl hydrogen peroxide, N-bromosuccinimide, hydrogen iodide, and phenol were carried out. The derivatives of carotol were designed to explore the significance of some structural modifications in relation to antioxidant activities. The structures of major compounds and derivatives were confirmed on the basis of FT-IR, 1HNMR and 13CNMR spectroscopy. Antioxidant activity of carrot seed essential oil, various extracts and isolated compounds were tested by in vitro models involving 2, 2-diphenyl-1-picrylhydrazyl (DPPH•), hydroxyl (OH•), nitric oxide (NO•), superoxide radical scavenging methods and ferric reducing antioxidant power assay (FRAP). Chemical transformations of major isolated compound carotol were carried out, and antioxidant activity of all compounds was undertaken. The major sesquiterpenoidcarotol isolated from carrot seed essential oil showed the highest antioxidant activity in all the methods. The methanol extract showed higher antioxidant potential as compared to carrot seed essential oil, hexane, and dichloromethane extracts.

Keywords: antioxidant, carotol, carrot, DPPH

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Authors: Prativa Das, Lay Poh Tan

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Three-dimensional microenvironment is a need to study the event cascades of liver fibrosis in vitro. Electrospun nanofibers modified with essential extracellular matrix proteins can closely mimic the random fibrous structure of native liver extracellular matrix (ECM). In this study, we fabricate a series of 3D electrospun scaffolds by wet electrospinning process modified with different ratios of collagen-I to fibronectin to achieve optimized distribution of these two ECM proteins on the fiber surface. A ratio of 3:1 of collagen-I to fibronectin was found to be optimum for surface modification of electrospun poly(lactic-co-glycolic acid) (PLGA) fibers by chemisorption process. In 3:1 collagen-I to fibronectin modified scaffolds the total protein content increased by ~2 fold compared to collagen-I modified and ~1.5 fold compared to 1:1/9:1 collagen-I to fibronectin modified scaffolds. We have cultured LX-2 cells on this scaffold over 14 days and found that LX-2 cells acquired more quiescent phenotype throughout the culture period and shown significantly lower expression of alpha smooth muscle actin and collagen-I. Thus, this system can be used as a model to study liver fibrosis by using different fibrogenic mediators in vitro.

Keywords: electrospinning, collagen-I and fibronectin, surface modification of fiber, LX-2 cells, liver fibrosis

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Authors: Münüre Tanur Erkoyuncu, Mustafa Yorgancılar

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Crop improvement through genetic engineering depends on effective and reproducible plant regeneration systems. Immature embryos are the most widely used explant source for in vitro regeneration in barley (Hordeum vulgare L.). However, immature embryos require the continuous growth of donor plants and the suitable stage for their culture is also certainly limited. On the other hand, mature embryos can be procured and stored easily; they can be studied throughout the year. In this study, an effective callus induction and plant regeneration were aimed to develop from mature embryos of different barley genotypes. The effect of medium (MS₁ and MS₂), auxin type (2,4-D, dicamba, picloram and 2,4,5-T) and concentrations (2, 4, 6 mg/l) on callus formation and effect of cytokinin type (TDZ, BAP) and concentrations (0.2, 0.5, 1.0 mg/l) on green plant regeneration were evaluated in mature embryo culture of barley. Callus and shoot formation was successful for all genotypes. By depending on genotype, MS₁ is the best medium, 4 mg/l dicamba is the best growth regulator in the callus induction and MS₁ is the best medium, 1 mg/l BAP is the best growth regulator in the shoot formation were determined.

Keywords: barley, callus, embryo culture, mature embryo

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Authors: Payman A. A. Zibari, Mosleh M. S. Duhoky

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These experiments were conducted at Tissue Culture Laboratory/ Faculty of Agriculture / University of Duhok during the period from January 2011 to May 2013. The objectives of this study were to study the effects of interaction between colchcine concentrations and treatment time duration of Creps capilaris (with and without 2B chromosome) on chromosome polyploidy during fifteen passages until regeneration of plants from the callus. Data showed that high percentage of chromosome polyploidy approximately can be obtained from high concentration of colchicin and long time of duration.

Keywords: polyploidy, Crepis capilaris, colchicine, B chromosome

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Authors: Adel S. El-Azab, Alaa A. M. Abdel-Aziz, Ibrahim A. Al-Suwaidan, Amer M. Alanazi

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A novel series of 2,3,6-trisubstitute quinazolinone were designed, synthesized, and evaluated for their in-vitro antitumor activity. 3 (Benzylideneamino)-6-chloro-2-p-tolylquinazolin-4(3H)-One, 2-[(4-oxo-3-phenethyl-3,4-dihydroquinazolin-2-yl)thio]-N-(3,4;5-trimethoxyphenyl) acetamide and 3-(3-benzyl-6-methyl-4-oxo-3, 4-dihydroquinazolin-2-ylthio)-N-(3,4,5-trimethoxyphenyl) propanamide have shown amazing broad spectrum antitumor activity with mean GI50; 15.8, 3.16, and 7.4 μM respectively compared to known Quinazoline Derivatives antitumor drug 5-FU mean GI50=22.6 μM.

Keywords: quinazoline derivatives, in vitro antitumor, synthesis, 5-FU, NCI

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Authors: Madalin Enache, Simona Neagu, Roxana Cojoc, Ioana Gomoiu, Delia Ionela Dobre, Ancuta Roxana Trifoi

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Various types of halophilic and halotolerant microorganisms able to be cultivated in laboratory on culture media with a wide range of sodium chloride content are isolated from several salted environments. The extracellular enzymes of these microorganisms showed the enzymatic activity in these spectrums of salinity thus being attractive for several biotechnological processes developed at high ionic strength. In present work, a number of amylase, protease, esterase, lipase, cellulase, pectinase, xilanases and innulinase were identified for more than 50th bacterial strains isolated from water samples and sapropelic mud from four saline and hypersaline lakes located in Romanian plain. On the other hand, the cellulase and pectinase activity were also detected in some halotolerant microorganisms isolated from secondary agricultural flow of grapes processing. The preliminary data revealed that from totally tested strains seven harbor proteases activity, eight amylase activity, four for esterase and another four for lipase, three for pectinase and for one strain were identified either cellulase or pectinase activity. There were no identified enzymes able to hydrolase innulin added to culture media. Several strains isolated from sapropelic mud showed multiple extracellular enzymatic activities, namely three strains harbor three activities and another seven harbor two activities. The data revealed that amylase and protease activities were frequently detected if compare with other tested enzymes. In the case of pectinase were investigated, their ability to be used for increasing resveratrol recovery from material resulted after grapes processing. In this way, the resulted material from grapes processing was treated with microbial supernatant for several times (two, four and 24 hours) and the content of resveratrol was detected by High Performance Liquid Chromatography method (HPLC). The preliminary data revealed some positive results of this treatment.

Keywords: halophilic microorganisms, enzymes, pectinase, salinity

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Authors: Biswabandhu Bankura, Srikanta Guria, Madhusudan Das

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Purpose: Thyroid peroxidase (TPO) is the key enzyme in the biosynthesis of thyroid hormones. We aimed to identify the spectrum of mutations in the TPO gene leading to hypothyroidism in the population of West Bengal to establish the genetic etiology of the disease. Methods: 200 hypothyroid patients (case) and their corresponding sex and age matched 200 normal individuals (control) were screened depending on their clinical manifestations. Genomic DNA was isolated from peripheral blood samples and TPO gene (Exon 7 to Exon 14) was amplified by PCR. The PCR products were subjected to sequencing to identify mutations. Results: Single nucleotide changes such as Glu 641 Lys, Asp 668 Asn, Thr 725 Pro, Asp 620 Asn, Ser 398 Thr, and Ala 373 Ser were found. Changes in the TPO were assayed in vitro to compare mutant and wild-type activities. Five mutants were enzymatically inactive in the guaiacol and iodide assays. This is a strong indication that the mutations are present at crucial positions of the TPO gene, resulting in inactivated TPO. Key Findings: The results of this study may help to develop a genetic screening protocol for goiter and hypothyroidism in the population of West Bengal.

Keywords: thyroid peroxidase, hypothyroidism, mutation, in vitro assay, transfection

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Authors: Biswabandhu Bankura, Srikanta Guria, Madhusudan Das

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Purpose: Thyroid peroxidase (TPO) is the key enzyme in the biosynthesis of thyroid hormones. We aimed to identify the spectrum of mutations in the TPO gene leading to hypothyroidism in the population of West Bengal to establish the genetic etiology of the disease. Methods: 200 hypothyroid patients (case) and their corresponding sex and age matched 200 normal individuals (control) were screened depending on their clinical manifestations. Genomic DNA was isolated from peripheral blood samples and TPO gene (Exon 7 to Exon 14) was amplified by PCR. The PCR products were subjected to sequencing to identify mutations. Results: Single nucleotide changes such as Glu 641 Lys, Asp 668 Asn, Thr 725 Pro, Asp 620 Asn, Ser 398 Thr, and Ala 373 Ser were found. Changes in the TPO were assayed in vitro to compare mutant and wild-type activities. Five mutants were enzymatically inactive in the guaiacol and iodide assays. This is a strong indication that the mutations are present at crucial positions of the TPO gene, resulting in inactivated TPO. Key Findings: The results of this study may help to develop a genetic screening protocol for goiter and hypothyroidism in the population of West Bengal.

Keywords: thyroid peroxidase, hypothyroidism, mutation, in vitro assay, transfection

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Authors: Recep Kesli, Merih Simsek, Cengiz Demir, Onur Turkyilmaz

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Objective: Goal of this study was to determine the antibiotic resistance of Staphylococcus aureus (S. aureus) and Methicillin resistant staphylococcus aureus (MRSA) strains isolated at Medical Microbiology Laboratory of ANS Application and Research Hospital, Afyon Kocatepe University, Turkey. Methods: S. aureus strains isolated between October 2012 and September 2016, from various clinical specimens were evaluated retrospectively. S. aureus strains were identified by both the conventional methods and automated identification system -VITEK 2 (bio-Mérieux, Marcy l’etoile, France), and Meticillin resistance was verified using oxacillin disk with disk-diffusion method. Antibiotic resistance testing was performed by Kirby-Bauer disc diffusion method according to CLSI criteria, and intermediate susceptible strains were considered as resistant. Results: Seven hundred S.aureus strains which were isolated from various clinical specimens were included in this study. These strains were mostly isolated from blood culture, tissue, wounds and bronchial aspiration. All of 306 (43,7%) were oxacillin resistant. While all the S.aureus strains were found to be susceptible to vancomycin, teicoplanin, daptomycin and linezolid, 38 (9.6 %), 77 (19.5 %), 116 (29.4 %), 152 (38.6 %) and 28 (7.1 %) were found to be resistant aganist to clindamycin, erythromycin, gentamicin, tetracycline and sulfamethoxazole/trimethoprim, retrospectively. Conclusions: Comparing to the Methicillin sensitive staphylococcus aureus (MSSA) strains, increased resistance rates of, trimethoprim-sulfamethoxazole, clindamycin, erythromycin, gentamicin, and tetracycline were observed among the MRSA strains. In this study, the most effective antibiotic on the total of strains was found to be trimethoprim-sulfamethoxazole, the least effective antibiotic on the total of strains was found to be tetracycline.

Keywords: antibiotic resistance, MRSA, Staphylococcus aureus, VITEK 2

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Authors: Nurlaili Syafar Wulan, Almurdi, Suprianto Kosno

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

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

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Authors: Alsulami H., Alghamdi N., Alasker A., Almohen N., Shome D.

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Most conventional chemotherapeutic agents, which are used for the treatment of cancers, not only eradicate cancer cells but also affect normal hematopoietic Stem cells (HSCs) that lead to severe pancytopenia during treatment. Therefore, a need exists for distinct approaches to treat cancer without or with minimum effect on normal HSCs. Parthenolide (PTL), a herbal product occurring naturally in the plant Feverfew, is a potential chemotherapeutic agent for the treatment of many cancers, such as acute myeloid leukemia (AML) and chronic lymphocytic leukemia (CLL). In this study we investigated the effect of different PTL concentrations on the viability of normal HSCs and also on the ability of these cells to form colonies after they have been treated with PTL in vitro. Methods: Normal 24 bone marrow and cord blood samples were included in this study after obtaining informed consent. The mononuclear cells were isolated using density gradient separation. Cells were cultured with different PTL concentrations for 24 hours. Post-culture cell viability was assessed using 7ADD in a flow cytometry-based test. In addition, a colony-forming unit assay (CFU) was carried out to assess the effect of PTL on HSCs. the expression of NFҝB was also assessed using a PE-labeled anti-pNFκBP65 antibody. Results: This study showed that there was no statistically significant difference in the percentage of cell death between untreated and PTL-treated cells with 5 μM PTL (p = 0.7), 10 μM PTL (p = 0.4) and 25 μM (p = 0.09) respectively. However, at higher doses, PTL caused a significant increase in the percentage of cell death. These results were significant when compared to untreated control (p < 0.001). The response of cord blood cells (n=4), on the other hand, was slightly different from that for bone marrow cells in that the percentage of cell death was significant at 100 μM PTL. Therefore, cord blood cells seemed more resistant than bone marrow cells. Conclusion: At concentrations ≤25 μM, PTL has a minimum or no effect on HSCs in vitro. Cord blood HSCs are more resistant to PTL compared to bone marrow HSCs.

Keywords: stem cell, parthenolide, ALL, NFKB

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Authors: Christelle E. Chua, Alicia L. Ho

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The development of a panel of differential gene expression signatures has been of interest in the field of biomarker discovery for radiation exposure. In the absence of the availability of exposed human subjects, lymphocyte cell lines have often been used as a surrogate to human whole blood, when performing ex vivo irradiation studies. The extent of variation between different lymphocyte cell lines is currently unclear, especially with regard to the expression of extracellular miRNA. This study compares the expression profile of extracellular miRNA isolated from different lymphocyte cell lines. It also compares the profile of miRNA obtained when different exosome isolation kits are used. Lymphocyte cell lines were created using lymphocytes isolated from healthy adult males of similar racial descent (Chinese American and Chinese Singaporean) and immortalised with Epstein-Barr virus. The cell lines were cultured in exosome-free cell culture media for 72h and the cell culture supernatant was removed for exosome isolation. Two exosome isolation kits were used. Total exosome isolation reagent (TEIR, ThermoFisher) is a polyethylene glycol (PEG)-based exosome precipitation kit, while ExoSpin (ES, Cell Guidance Systems) is a PEG-based exosome precipitation kit that includes an additional size exclusion chromatography step. miRNA from the isolated exosomes were isolated using miRNEASY minikit (Qiagen) and analysed using nCounter miRNA assay (Nanostring). Principal component analysis (PCA) results suggested that the overall extracellular miRNA expression profile differed between the lymphocyte cell line originating from the Chinese American donor and the cell line originating from the Chinese Singaporean donor. As the gender, age and racial origins of both donors are similar, this may suggest that there are other genetic or epigenetic differences that account for the variation in extracellular miRNA gene expression in lymphocyte cell lines. However, statistical analysis showed that only 3 miRNA genes had a fold difference > 2 at p < 0.05, suggesting that the differences may not be of that great a significance as to impact overall conclusions drawn from different cell lines. Subsequent analysis using cell lines from other donors will give further insight into the reproducibility of results when difference cell lines are used. PCA results also suggested that the method of exosome isolation impacted the expression profile. 107 miRNA had a fold difference > 2 at p < 0.05. This suggests that the inclusion of an additional size exclusion chromatography step altered the subset of the extracellular vesicles that were isolated. In conclusion, these results suggest that extracellular miRNA can be isolated and analysed from exosomes derived from lymphocyte cell lines. However, care must be taken in the choice of cell line and method of exosome isolation used.

Keywords: biomarker, extracellular miRNA, isolation methods, lymphocyte cell line

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Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

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Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

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Authors: Muhammad Aasim, Mehmet Karataş, Fatih Erci, Şeyma Bakırcı, Ecenur Korkmaz, Burak Kahveci

Abstract:

Water hyssop (Bacopa monnieri L. Wettst.) is an important medicinal aquatic/semi aquatic plant native to India where it is used in traditional medicinal system. The plant contains bioactive compounds mainly Bacosides which are the main ingridient of commercial drug available as memory enhancer tonic. The local name of water hyssop is Brahmi and brahmi based drugs are available against for curing chronic diseases and disorders Alzheimer’s disease, anxiety, asthma, cancer, mental illness, respiratory ailments, and stomach ulcers. The plant is not a cultivated plant and collection of plant from nature make palnt threatened to endangered. On the other hand, low seed viability and availability make it difficult to propagate plant through traditional techniques. In recent years, plant tissue culture techniques have been employed to propagate plant for its conservation and production for continuous availability of secondary metabolites. On the other hand, application of nanoparticles has been reported for increasing biomass, in vitro regeneration and secondary metabolites production. In this study, silver nanoparticles (AgNPs) were applied at the rate of 2, 4, 6, 8 and 10 ppm to Murashihe and Skoog (MS) medium supplemented with 1.0 mg/l Benzylaminopurine (BAP), 3.0% sucrose and 0.7% agar. Leaf explants of water hyssop were cultured on AgNPs containing medium. Shoot induction from leaf explants were relatively slow compared to medium without AgNPs. Multiple shoot induction was recorded after 3-4 weeks of culture comapred to control that occured within 10 days. Regenerated shoots were rooted successfully on MS medium supplemented with 1.0 mg/l IBA and acclimatized in the aquariums for further studies.

Keywords: Water hyssop, Silver nanoparticles, In vitro, Regeneration, Secondary metabolites

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Authors: Haba Hamada, Lavaud Cathrine, Benkhaled Mohammed

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The Silene genus is the most representative of the caryophyllaceae family for their rich content in secondary metabolites; saponins, flavonoids and flavonoids glycosides, phytoecdysones, oligosaccharides have been isolated and identified. The Silene genus represented by about 700 species in the temrerate region of the word, the main concentration of spcies is Europe, Asia and North Africa. Three known compounds 1-3 were isolated from the aerial parts of Silene arenarioides Desf. by using different chromatographic methods. The structures of the isolated compounds were determined as stigmasterolglycoside, Soyacerebroside, maltol glycoside. The structures of the isolated compounds were determined by using the NMR (1H-NMR, 13C-NMR, COSY, HSQC, and HMBC) techniques and mass spectroscopy. The antimicrobial and antioxydant activities of the different extracts and compound have been reported.

Keywords: caryophyllaceae, flavonoids, saponosids, flavonoids glycosides

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Authors: M. Raissy, M. Shahrani

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The present study was done to evaluate the presence of tetracycline resistance genes in Lactococcus garvieae isolated from cultured rainbow trout, West Iran. The isolates were examined for antimicrobial resistance using disc diffusion method. Of the 49 strains tested, 19 were resistant to tetracycline (38.7%), 32 to enrofloxacin (65.3%), 21 to erythromycin (42.8%), 20 to chloramphenicol and trimetoprim-sulfamethoxazole (40.8%). The strains were then characterized for their genotypic resistance profiles. The results revealed that all 49 isolates contained at least one of the tetracycline resistance genes. Tet (A) was found in 89.4% of tetracycline resistant isolates and the frequency of other gene were as follow: tet (E) 42.1%, tet (B) 47.3%, tet (D) 15.7%, tet (L) 26.3%, tet (K) 52.6%, tet (G) 36.8%, tet (34) 21%, tet (S) 63.1%, tet (C) 57.8%, tet (M) 73.6%, tet (O) 42.1%. The results revealed high levels of antibiotic resistance in L. garvieae strains which is a potential danger for trout culture as well as for public health.

Keywords: Lactococcus garvieae, tetracycline resistance genes, rainbow trout, antimicrobial resistance

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Authors: Krishna Poudel, Tahar Katuwal, Sujan Karki

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A rapid propagation and acclimatization method of large cardamom was optimized in this study. Sprouted rhizome buds were collected. The excised rhizome bud explants were cultured on semi solid culture media. The explants were cultured on Murashige and Skoog’s (MS) medium supplemented with different concentration and combinations of BAP (6-Benzyl-amino-purine) and IBA (Indole-3-butyric acid) for shoot and root induction. Explants cultured on MS basal medium supplemented with 1.0 mg/l BAP + 0.5 gm/l IBA showed the highest rate of shoot multiplication. In vitro shoots were rooted on to the half-strength MS basal media supplemented with 0.5 mg/l IBA. Rooted shoots were transplanted in the screen house for hardening process. These hardened plants were subsequently shifted into the netted nursery for further multiplication process.

Keywords: concentration, explants, hardening, rhizome

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Authors: Ezeonu, Chukwuma Stephen, Onwurah, Ikechukwu Noel Emmanuel

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Pure strains of Aspergillus fumigatus (AF), aspergillus niger (AN), aspergillus oryzae (AO), trichophyton mentagrophyte (TM), trichophyton rubrum (TR) and Trichophyton soudanense (TS) were isolated from decomposing rice husk. Freshly processed rice husk in Mandle’s medium were heat pre-treated using an autoclave at 121oC for 20 minutes. The isolated fungi as monoculture and di-culture combinations were inoculated into each of the pre-treated rice husk with the exception of two controls. Seven days hydrolysis was followed by estimation of carbohydrate, reducing sugar and non-reducing sugar. Fungal treated rice husks were left to ferment for 7 days with introduction of both baker’s and palm wine yeast. The result obtained in the work gave the highest carbohydrate (20.53 ± 2.73 %) from rice husks treated with TS + TR di-culture. The highest soluble reducing sugar (2.66 ± 0.14 %) was obtained from rice husk treated with TM. The highest soluble nonreducing sugar (18.08 ± 2.61 %) was from AF. The introduction of yeasts from palm wine gave the highest bio-ethanol (12.82 ± 0.39 %) from AO. The highest bio-ethanol (6.60 ± 0.10 %) from baker's yeast fermentation was in AO + TS treated rice husk. There was increased availability of sugar and moderate yield of bio-ethanol, especially from palm wine yeast.

Keywords: fungi, rice husk, carbohydrate, reducing sugar, non-reducing sugar, ethanol, fermentation

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Authors: Chaoran Liu

Abstract:

Bioactive polysaccharides and polysaccharide-protein complex derived from mushrooms and fungi have a wide range of immunomodulatory activity with low side-effects and have therefore the potential to be developed as an adjuvant in cancer therapies. Mushrooms sclerotium is rich in polysaccharides and the polysaccharides isolated from the sclerotium of Polyporus rhinocerus have shown potent in vivo and in vitro immunomodulatory effects. Macrophages are considered to be an important component of the innate immune response against bacterial infection and cancer. To better understanding the immunomodulatory effects and its underlying mechanisms of sclerotial water-soluble polysaccharides extracted from P. rhinocerus on macrophages, the objectives of this study are to purify the water-soluble novel sclerotial polysaccharides and to characterize the structure and properties as well as to study the detailed molecular mechanisms of the in vitro immunomodulating effects in murine macrophages. The hot water-soluble fraction PRW from the sclerotium of P. rhinocerus was obtained using solvent extraction. PRW was further fractionated by membrane ultrafiltration to a give a fraction (PRW1) with molecular mass less than 50 kDa. PRW1 was characterized to be a polysaccharide-protein complex composed of 45.7% polysaccharide and 44.2% protein. The chemical structure of the carbohydrate moiety of PRW1 was elucidated by GC and FTIR to be mainly beta-D-glucan with trace amount of galactose and mannose. The immunomodulatory effects of PRW1 on murine RAW 264.7 macrophages were demonstrated in terms of the increase in nitric oxide production and cytokine production. Mechanistically, PRW1 initiates ERK phosphorylation to activate macrophages within 15 min and significantly improves the expression level of inducible NOS (iNOS) from 6 h after treatment. In summary, this study indicates that PRW1 is a potent immunomodulatory agent for macrophages and suggests that mushroom sclerotia from Polyporus rhinocerus requires for further investigation in cancer research.

Keywords: Polyporus rhinocerus, mushroom sclerotia, Polysaccharide-Protein Complex, macrophage activation

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Authors: Nunthawan Nowwarote, Waleerat Sukarawan, Prasit Pavasant, Thanaphum Osathanon

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Interleukin 6 (IL-6) is a multifunctional cytokine, regulating various biological responses in several tissues. A Recent study shows that IL-6 plays a role in stemness maintenance in stem cells isolated from human exfoliated deciduous teeth (SHEDs). However, the role of IL-6 on cell differentiation in SHEDs remains unknown. The present study investigated the effect of IL-6 on SHEDs differentiation. Cells were isolated from dental pulp tissues of human deciduous teeth. Flow cytometry was used to determined mesenchymal stem cell marker expression, and the multipotential differentiation (osteogenic, adipogenic and neurogenic lineage ) was also determined. The mRNA was determined using real-time quantitative polymerase chain reaction, and the phenotypes were confirmed by chemical and immunofluorescence staining. Results demonstrated that SHEDs expressed CD44, CD73, CD90, CD105 but not CD45. Further, the up-regulation of osteogenic, adipogenic and neurogenic marker genes was observed upon maintaining cells in osteogenic, adipogenic and neurogenic induction medium, respectively. The addition of IL-6 induced osteogenic by up-regulated osteogenic marker gene also increased in vitro mineralization. Under neurogenic medium supplement with IL-6, up-regulated neurogenic marker. Whereas, an addition of IL-6 attenuated adipogenic differentiation by SHEDs. In conclusion, this evidence implies that IL-6 may participate in cells differentiation ability of SHEDs.

Keywords: SHEDs, IL-6, cell differentiations, dental pulp

Procedia PDF Downloads 142