Search results for: mosaic antigen

Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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Protein apoferritin seems to be a very promising structure for use as a nanocarrier. It is prepared from intracellular ferritin protein naturally found in most organisms. The role of ferritin proteins is to store and transport ferrous ions. Apoferritin is a hollow protein cage without ferrous ions that can be prepared from ferritin by reduction with thioglycolic acid or dithionite. The structure of apoferritin is composed of 24 protein subunits, creating a sphere with 12 nm in diameter. The inner cavity has a diameter of 8 nm. The drug encapsulation process is based on the response of apoferritin structure to the pH changes of surrounding solution. In low pH, apoferritin is disassembled into individual subunits and its structure is “opened”. It can then be mixed with any desired cytotoxic drug and after adjustment of pH back to neutral the subunits are reconnected again and the drug is encapsulated within the apoferritin particles. Excess drug molecules can be removed by dialysis. The receptors for apoferritin, SCARA5 and TfR1 can be found in the membrane of both healthy and cancer cells. To enhance the specific targeting of apoferritin nanocarrier, it is possible to modify its surface with targeting moieties, such as antibodies. To ensure sterically correct complex, we used a a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (ApoDox) was coated either with gold nanoparticles (ApoDox-Nano) or gold (III) chloride hydrate reduced with sodium borohydride (ApoDox-HAu). The applied amount of gold in form of gold (III) chloride hydrate was 10 times higher than in the case of gold nanoparticles. However, after removal of the excess unbound ions by electrophoretic separation, the concentration of gold on the surface of apoferritin was only 6 times higher for ApoDox-HAu in comparison with ApoDox-Nano. Moreover, the reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties (excitation maximum at 480 nm with emission maximum at 600 nm) and thus its biological activity. Fluorescent properties of ApoDox-Nano were similar to the unmodified ApoDox, therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, we used ELISA-like method with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, we applied ApoDox without targeting antibodies and ApoDox-Nano modified with targeting antibodies (human IgG antibodies). The amount of unmodified ApoDox on antigen after incubation and subsequent rinsing with water was 5 times lower than in the case of ApoDox-Nano modified with targeting antibodies. The modification of non-gold ApoDox with antibodies caused no change in its targeting properties. It can therefore be concluded that the demonstrated procedure allows us to create nanocarrier with enhanced targeting properties, suitable for nanomedicine.

Keywords: apoferritin, doxorubicin, nanocarrier, targeting antibodies

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Authors: Khadidja Rahmani, Nahla Bouaziz

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Algeria is known for its rich heritage. It owns an immense historical heritage with a universal reputation. Unfortunately, this wealth is withered because of abundance. This research focuses on the Ksourian model, which constitutes a large portion of this wealth. In fact, the Ksourian model is not just a witness to a great part of history or a vernacular culture, but also it includes a panoply of assets in terms of energetic efficiency. In this context, the purpose of our work is to evaluate the performance of the old techniques which are derived from the Ksourian model , and that using the simulation tools. The proposed method is decomposed in two steps; the first consists of isolate and reintroduce each device into a basic model, then run a simulation series on acquired models. And this in order to test the contribution of each of these dialectal processes. In another scale of development, the second step consists of aggregating all these processes in an aboriginal model, then we restart the simulation, to see what it will give this mosaic on the environmental and energetic plan .The model chosen for this study is one of the ksar units of Knadsa city of Bechar (Algeria). This study does not only show the ingenuity of our ancestors in their know-how, and their adapting power to the aridity of the climate, but also proves that their conceptions subscribe in the current concerns of energy efficiency, and respond to the requirements of sustainable development.

Keywords: dialectal processes, energy efficiency, evaluation, Ksourian model, simulation tools

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Authors: Vanessa Roulet, Marc Turini, Juliane Hey, Stéphanie Bord-Romeu, Emilie Bonzom, Mahmoud Badawi, Mohammed-Amine Chakir, Valérie Simon, Vanessa Viotti, Jérémie Gautier, Françoise Le Boulaire, Catherine Coignard, Claire Vincent, Sandrine Greaume, Isabelle Voisin

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Background: Beckman Coulter, Inc. has recently developed fully automated assays for the detection of HBsAg on a new immunoassay platform. The objective of this European multicenter study was to evaluate the performance of the ACCESS HBsAg and ACCESS HBsAg Confirmatory assays† on the recently CE-marked DxI 9000 ACCESS Immunoassay Analyzer. Methods: The clinical specificity of the ACCESS HBsAg and HBsAg Confirmatory assays was determined using HBsAg-negative samples from blood donors and hospitalized patients. The clinical sensitivity was determined using presumed HBsAg-positive samples. Sample HBsAg status was determined using a CE-marked HBsAg assay (Abbott ARCHITECT HBsAg Qualitative II, Roche Elecsys HBsAg II, or Abbott PRISM HBsAg assay) and a CE-marked HBsAg confirmatory assay (Abbott ARCHITECT HBsAg Qualitative II Confirmatory or Abbott PRISM HBsAg Confirmatory assay) according to manufacturer package inserts and pre-determined testing algorithms. False initial reactive rate was determined on fresh hospitalized patient samples. The sensitivity for the early detection of HBV infection was assessed internally on thirty (30) seroconversion panels. Results: Clinical specificity was 99.95% (95% CI, 99.86 – 99.99%) on 6047 blood donors and 99.71% (95%CI, 99.15 – 99.94%) on 1023 hospitalized patient samples. A total of six (6) samples were found false positive with the ACCESS HBsAg assay. None were confirmed for the presence of HBsAg with the ACCESS HBsAg Confirmatory assay. Clinical sensitivity on 455 HBsAg-positive samples was 100.00% (95% CI, 99.19 – 100.00%) for the ACCESS HBsAg assay alone and for the ACCESS HBsAg Confirmatory assay. The false initial reactive rate on 821 fresh hospitalized patient samples was 0.24% (95% CI, 0.03 – 0.87%). Results obtained on 30 seroconversion panels demonstrated that the ACCESS HBsAg assay had equivalent sensitivity performances compared to the Abbott ARCHITECT HBsAg Qualitative II assay with an average bleed difference since first reactive bleed of 0.13. All bleeds found reactive in ACCESS HBsAg assay were confirmed in ACCESS HBsAg Confirmatory assay. Conclusion: The newly developed ACCESS HBsAg and ACCESS HBsAg Confirmatory assays from Beckman Coulter have demonstrated high clinical sensitivity and specificity, equivalent to currently marketed HBsAg assays, as well as a low false initial reactive rate. †Pending achievement of CE compliance; not yet available for in vitro diagnostic use. 2023-11317 Beckman Coulter and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries. All other trademarks are the property of their respective owners.

Keywords: dxi 9000 access immunoassay analyzer, hbsag, hbv, hepatitis b surface antigen, hepatitis b virus, immunoassay

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Authors: Satoshi Furukawa, Satomu Morita, Lisa Wingenfeld, Katsuji Nishi, Masahito Hitosugi

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We studied the expression of hypoxia-related antigens (e.g., cold-inducible antigens and apoptotic antigens) in the myocardium and the cerebellumthat were obtained from individuals after death by carbon monoxide or hypothermia. The immunohistochemistry results revealed that expression of cold-inducible RNA binding protein (CIRBP) and RNA-binding protein 3 (RBM3) may be associated with hpyothermic and the hypoxic conditions. The expression of CIRBP and RBM3 in the myocardium was different from their expression in the cerebellum, especially in the Purkinje cells. The results indicate that agonal duration influences antigen expression. In the hypothermic condition, the myocardium uses more ATP since the force of the excitation-contraction coupling of the myocardium increases by more than 400% when the experimental temperature is reduced from 35°C to 25°C. The results obtained in this study indicate that physicians should pay attention to the myocardium when cooling the patient’s body to protect the brain.

Keywords: carbon monoxide death, cerebellum, CIRBP, hypothermic death, myocardium, RBM3

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Authors: Mehmet Kale, Ramazan Sencan, Sibel Yavru, Ahmet Ak, Nuri Mamak, Sibel Hasırcıoglu, Mesih Kocamuftuoglu, Yakup Yıldırım, Hasbi Sait Saltık

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Bovine Papilloma Virus (BPV)-induced warts can cause mastitis, teat blindness, reduction of milk yield, udder deformities, and a difficulty in getting the teats into the milking machine. Especially, surgical operations cannot be performed in BPV-induced teat warts because of the increased sensitivity of the breast region and small-sized papillomas. Thus, there is a need to find new topical treatment methods. We have developed a pomade for treatment of BPV in cattle. The pomade is consists of lanoline, snakeskin (two special kind of snake), alcohol, vaseline, and ether. Firstly, we determined 46 cattle with teat warts. In the study, BPV antigen was detected in 28 cattle blood samples (61%) by ELISA. The pomade was applied to all BPV infected animals. The regression and recovery of warts were 100% in all animals. We advised using the pomade for treatment of BPV-induced warts in teats.

Keywords: bovine papilloma virus, pomade, teat, udder

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Authors: Y. Sarı Nayim, B. N. Nayim

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This research proposal includes the defining and evaluation of the semi-natural and cultural ecosystems at Bartın University main campus in Turkey in terms of landscape mosaics. The ecosystem mosaic of the main campus was divided into zones based on ecological classification technique. Based on the results from the study, it was found that 6 different ecosystem mosaics should be used as a base in the planning and design of the existing and future landscape planning of Kutlubeyyazıcılar campus. The first landscape zone involves the 'social areas'. These areas include yards, dining areas, recreational areas and lawn areas. The second landscape zone is 'main vehicle and pedestrian areas'. These areas include vehicle access to the campus landscape, moving in the campus with vehicles, parking and pedestrian walk ways. The third zone is 'landscape areas with high visual landscape quality'. These areas will be the places where attractive structural and plant landscape elements will be used. Fourth zone will be 'landscapes of building borders and their surroundings.' The fifth and important zone that should be survived in the future is 'Actual semi-natural forest and bush areas'. And the last zone is 'water landscape' which brings ecological value to landscape areas. While determining the most convenient areas in the planning and design of the campus, these landscape mosaics should be taken into consideration. This zoning will ensure that the campus landscape is protected and living spaces in the campus apart from the areas where human activities are carried out will be used properly.

Keywords: campus landscape planning and design, landscape ecology, landscape mosaics, Bartın

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Authors: Yefrem Yefremov

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In Kazakhstan, there are approximately 100,000 ethnic Koreans with the ethnonym "Koryo Saram". They are part of the global Korean diaspora around the world, deported to Kazakhstan by Stalin’s decree in 1937. Koryo Saram's diasporic identity is a composite of numerous identities based on a shared cultural heritage of the USSR and independent Kazakhstan and has mosaic character. The author has conducted a sociological survey to find out the main features of the identity of the Koryo Saram diaspora. The purpose of this paper is to depict the degree of ethnic, cultural, and diasporic identity of Koryo Saram and which effect on the preserving Korean diaspora in Kazakhstna do they have. The following elements impacting the above-mentioned identities were investigated in the survey: criteria by which Koryo Saram perceive themselves to be Korean, attitude of Koryo Saram to their ethnicity, degree of feeling of ethnocultural similarity between Koreans of Kazakhstan and Koreans of the Republic of Korea, degree of association of Koreans of Kazakhstan with other Koreans living in other CIS countries, degree of practicing Korean traditions Koryo Saram's attitudes towards interethnic marriages. The primary factor in defining the identity among the respondents is the factor of ethnic origin. Nationality is the second most significant component in establishing Koryo Saram’s identity. The maintenance of "Koreanness" of Koryo Sarams in the context of a multiethnic community, particularly in Kazakhstan, is based on genetic elements as well as the preservation of the culture. In conclusion, the high level of preserving Korean identity is being observed in the Korean Diaspora of Kazakhstan.

Keywords: diasporic identity, diaspora, ethnic identity, identity markers, korean diaspora, koreans of kazakhstan, koryo saram, multiethnicity

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Authors: Diana Serbezeanu, Ionela-Daniela Carja, Tachita Vlad-Bubulac, Sergiu Sova

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New phosphorus-containing monomers having methoxy end functional groups were prepared from methyl 4-hydroxybenzoate and two different dichlorides with phosphorus, namely phenyl phosphonic dichloride and phenyl dichlorophosphate. The structures of the monomers were confirmed by FTIR and NMR spectroscopy. The assignments for the 1H, 13C and 31P chemical shifts are based on 1D and 2D NMR homo- and heteronuclear correlations (H,H-COSY (Correlation Spectroscopy), H,C-HMQC (Heteronuclear Multiple Quantum Correlation and H,C-HMBC (Heteronuclear Multiple Bond Correlation)) and 31P-13C couplings. The monomers exhibited good solubility in common organic solvents. Dimethyl sulfoxide was to be a good solvent to grow crystals of considerable size which were investigated by X-ray analysis. One of these two new monomers presented thermotropic liquid crystalline behaviour, as revealed by differential scanning calorimetry (DSC), polarized light microscopy (PLM) and X-ray diffraction (XRD). The transition temperature from crystal to liquid crystalline state (K→LC) was 143°C and from the LC to isotropic state (LC→I) was 167°C. Upon heating, bis(4-(methoxycarbonyl)phenyl formed fine textures, difficult to be ascribed to smectic or nematic phases. Upon cooling from the isotropic state, bis(4-(methoxycarbonyl)phenyl exhibited a mosaic-type texture. X-ray diffraction measurements at small angles (SAXS) of bis(4-(methoxycarbonyl)phenyl showed two peaks at 1.8 Å and 3.5 Å, respectively suggesting organization at supramolecular level.

Keywords: phosphorus-containing monomers, polarized light microscopy, structure investigation, thermotropic liquid crystalline properties

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Authors: Andreas Latz, Daniela Heinz, Fatima Hashemi, Melek Baygül

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Introduction: Feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and feline coronavirus (FCoV) are serious infectious diseases affecting cats worldwide. Transmission of these viruses occurs primarily through close contact with infected cats (via saliva, nasal secretions, faeces, etc.). FeLV, FIV, and FCoV infections can occur in combination and are expressed in similar clinical symptoms. Diagnosis can therefore be challenging: Symptoms are variable and often non-specific. Sick cats show very similar clinical symptoms: apathy, anorexia, fever, immunodeficiency syndrome, anemia, etc. Sample volume for small companion animals for diagnostic purposes can be challenging to collect. In addition, multiplex diagnosis of diseases can contribute to an easier, cheaper, and faster workflow in the lab as well as to the better differential diagnosis of diseases. For this reason, we wanted to develop a new diagnostic tool that utilizes less sample volume, reagents, and consumables than multiplesingleplex ELISA assays Methods: The Multiplier from Dynextechonogies (USA) has been used as platform to develop a Multiplex diagnostic tool for the detection of antibodies against FIV and FCoV/FIP and antigens for FeLV. Multiplex diagnostics. The Dynex®Multiplier®is a fully automated chemiluminescence immunoassay analyzer that significantly simplifies laboratory workflow. The Multiplier®ease-of-use reduces pre-analytical steps by combining the power of efficiently multiplexing multiple assays with the simplicity of automated microplate processing. Plastic beads have been coated with antigens for FIV and FCoV/FIP, as well as antibodies for FeLV. Feline blood samples are incubated with the beads. Read out of results is performed via chemiluminescence Results: Bead coating was optimized for each individual antigen or capture antibody and then combined in the multiplex diagnostic tool. HRP: Antibody conjugates for FIV and FCoV antibodies, as well as detection antibodies for FeLV antigen, have been adjusted and mixed. 3 individual prototyple batches of the assay have been produced. We analyzed for each disease 50 well defined positive and negative samples. Results show an excellent diagnostic performance of the simultaneous detection of antibodies or antigens against these feline diseases in a fully automated system. A 100% concordance with singleplex methods like ELISA or IFA can be observed. Intra- and Inter-Assays showed a high precision of the test with CV values below 10% for each individual bead. Accelerated stability testing indicate a shelf life of at least 1 year. Conclusion: The new tool can be used for multiplex diagnostics of the most important feline infectious diseases. Only a very small sample volume is required. Fully automation results in a very convenient and fast method for diagnosing animal diseases.With its large specimen capacity to process over 576 samples per 8-hours shift and provide up to 3,456 results, very high laboratory productivity and reagent savings can be achieved.

Keywords: Multiplex, FIV, FeLV, FCoV, FIP

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Authors: Ibrahim Aly, Waleed Elawamy, Hanan Taher, Amira Matar

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Schistosomiasis is infection with blood flukes of the genus Schistosoma, which are acquired trans-cutaneously by swimming or wading in contaminated freshwater. The present study was proposed to produce ultra-sensitive, field-friendly high-throughput rapid immunochromatography diagnostic device for accurate detection of asymptomatic parasite carriers in schistosomiasis pre-elimination settings.For assessing diagnostic potential of rapid device, 50 blood samples from patients with schistosomiasis mansoni, 29 other proven parasitic diseases and 25 blood samples as negative control were from healthy individuals were used. The sensitivity of Quantitative antigen-capture nano-ELISAwas 82 %, and specificity was 87.1 %, where the sensitivity of Nano Dot- ELISA was 86 % and specificity was 90.7 %. The sensitivity of diagnostic device was 78 % and specificity was 85.2 %, with PPV and NPV of 86.2 % and 83.1 %, respectively.The Point of care device resulted in a good performance for the diagnosis of low-intensity infections, it was able to identify 19 out of 25 (76 %) individuals with ⩽7 eggs, 10 out of 14 individuals (71.4 %) with 11–99 eggs and 100 % of individuals with 100–399 eggs.

Keywords: schistosomiasis, immunochromatography, naon-dot-ELISa, diagnostis device

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Authors: Farrah Putri Salmanida, Mei-Li Wu, Rika Wahyuningtyas, Wen-Bin Chung, Hso-Chi Chaung, Ko-Tung Chang

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Within the field of immunotherapy, oncolytic virotherapy (OVT) employs dual approaches that directly eliminate tumor cells while preserving healthy ones and indirectly reprogram the tumor microenvironment (TME) to elicit antitumor responses. Within the TME, tumor associated macrophages (TAMs) manifest characteristics akin to those of anti-inflammatory M2 macrophages, thus earning the designation of M2-like TAMs. In prior research, two antigens denoted as A1 (g6Ld10T) and A3 (ORF6L5), derived from a complete sequence of ORF5 with partial sequence of ORF6 in Porcine Reproductive and Respiratory Syndrome Virus Genotype 2 (PRRSV-2), demonstrated the capacity to repolarize M2-type porcine alveolar macrophages (PAMs) into M1 phenotypes. In this study, we sought for utilizing OVT strategies by introducing A1 or A3 on TAMs to endow them with the anti-tumor traits of M1 macrophages while retaining their capacity to target cancer cells. Upon exposing human THP-1-derived M2 macrophages to a cross-species test with 2 µg/ml of either A1 or A3 for 24 hours, real time PCR revealed that A3, but not A1, treated cells exhibited upregulated gene expressions of M1 markers (CCR7, IL-1ß, CCL2, Cox2, CD80). These cells reacted to virus-derived antigen, as evidenced by increased expression of pattern-recognition receptors TLR3, TLR7, and TLR9, subsequently providing feedback in the form of type I interferon responses like IFNAR1, IFN-ß, IRF3, IRF7, OAS1, Mx1, and ISG15. Through an MTT assay, only after 15 µg/ml of A3 treatment could the cell viability decrease, with a predicted IC50 of 16.96 µg/ml. Interestingly, A3 caused dose-dependent toxicity to a rat C6 glial cancer cell line even at doses as low as 2.5 µg/ml and reached its IC50 at 9.419 µg/ml. Using Annexin V/7AAD staining and PCR test, we deduced that a significant proportion of C6 cells were undergoing the early apoptosis phase predominantly through the intrinsic apoptosis cascade involving Bcl-2 family proteins. Following this stage, we conducted a test on A3’s repolarization ability, which revealed a significant rise in M1 gene expression markers, such as TNF, CD80, and IL-1ß, in M2-like TAMs generated in vitro from murine RAW264.7 macrophages grown with conditioned medium of 4T1 breast cancer cells. This was corroborated by the results of transcriptome analysis, which revealed that the primary subset among the top 10 to top 30 significantly upregulated differentially expressed genes (DEGs) dominantly consisted of M1 macrophages profiles, including Ccl3, Ccl4, Csf3, TNF, Bcl6b, Stc1, and Dusp2. Our findings unveiled the remarkable potential of the PRRSV-derived antigen A3 to repolarize macrophages while also being capable of selectively inducing apoptosis in cancerous cells. While further in vivo study is needed for A3, it holds promise as an adjuvant by its dual effects in cancer therapy modalities.

Keywords: cancer cell apoptosis, interferon responses, macrophage repolarization, recombinant protein

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Authors: T. Nalowa, L. Foncha, S. Eposi

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Prostate enlargement, prostate cancer are major global health problems affecting many men as they advance in age. It is highly recommended to encourage older men to get Prostate Specific Antigen test screening frequently. Conventional treatments like radiation, chemotherapy are associated with many side effects. And this situation is a call for concern. Traditional medicine is affordable, easily prepared with little or no side effects and it contains many phytochemicals. The study aims to find the cure for prostate cancer and prostate enlargement by extracting products from plant tissues of specific herbs to determine anti-inflammatory, anti-cancer, and anti-hematuria properties. Descriptive statistical analysis was applied to describe the data process. The commonly used method of preparation was extraction. Overall, 40 patients were classified based on their medical conditions on their underlying user report. Rural communities in Fako are rich sources of plants with medicinal properties. The used plants consequently provide basic information and aid to investigate the cure of prostate cancer and prostate enlargement, with great significance.

Keywords: cancer, enlargement, metastases, prostate

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Authors: Massimiliano Biagini, Marco Spinsanti, Gabriella De Angelis, Sara Tomei, Ilaria Ferlenghi, Maria Scarselli, Alessia Biolchi, Alessandro Muzzi, Brunella Brunelli, Silvana Savino, Marzia M. Giuliani, Isabel Delany, Paolo Costantino, Rino Rappuoli, Vega Masignani, Nathalie Norais

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The gram-negative bacterium Neisseria meningitidis serogroup B (MenB) is an exclusively human pathogen representing the major cause of meningitides and severe sepsis in infants and children but also in young adults. This pathogen is usually present in the 30% of healthy population that act as a reservoir, spreading it through saliva and respiratory fluids during coughing, sneezing, kissing. Among surface-exposed protein components of this diplococcus, factor H binding protein is a lipoprotein proved to be a protective antigen used as a component of the recently licensed Bexsero vaccine. fHbp is a highly variable meningococcal protein: to reflect its remarkable sequence variability, it has been classified in three variants (or two subfamilies), and with poor cross-protection among the different variants. Furthermore, the level of fHbp expression varies significantly among strains, and this has also been considered an important factor for predicting MenB strain susceptibility to anti-fHbp antisera. Different methods have been used to assess fHbp expression on meningococcal strains, however, all these methods use anti-fHbp antibodies, and for this reason, the results are affected by the different affinity that antibodies can have to different antigenic variants. To overcome the limitations of an antibody-based quantification, we developed a quantitative Mass Spectrometry (MS) approach. Selected Reaction Monitoring (SRM) recently emerged as a powerful MS tool for detecting and quantifying proteins in complex mixtures. SRM is based on the targeted detection of ProteoTypicPeptides (PTPs), which are unique signatures of a protein that can be easily detected and quantified by MS. This approach, proven to be highly sensitive, quantitatively accurate and highly reproducible, was used to quantify the absolute amount of fHbp antigen in total extracts derived from 105 clinical isolates, evenly distributed among the three main variant groups and selected to be representative of the fHbp circulating subvariants around the world. We extended the study at the genetic level investigating the correlation between the differential level of expression and polymorphisms present within the genes and their promoter sequences. The implications of fHbp expression on the susceptibility of the strain to killing by anti-fHbp antisera are also presented. To date this is the first comprehensive fHbp expression profiling in a large panel of Neisseria meningitidis clinical isolates driven by an antibody-independent MS-based methodology, opening the door to new applications in vaccine coverage prediction and reinforcing the molecular understanding of released vaccines.

Keywords: quantitative mass spectrometry, Neisseria meningitidis, vaccines, bexsero, molecular epidemiology

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Authors: Aleena Rena Onozuka, Patompong Satapornpong

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Introduction: In a Human Leukocyte Antigen (HLA)’s immune response, HLA alleles (HLA class I and class II) play a crucial role in fighting against pathogens. HLA-B*15:01 allele had a significant association with asymptomatic COVID-19 infection (p-value = 5.67 x 10-5 ; OR = 2.40 and 95% CI = 1.54 - 3.64). There was also a notable linkage between HLA-B*51:01 allele and critically ill patients with COVID-19 (p-value = 0.007 and OR = 3.38). This study has described the distribution of HLA marker alleles in Thais and sub-groups. Objective: We want to investigate the prevalence of HLA-B*15:01 and HLA-B*51:01 alleles in the Thai population. Materials and Methods: 200 healthy Thai population were included in this study from the College of Pharmacy, Rangsit University. HLA-B alleles were genotyped using the sequence-specific oligonucleotides process (PCR-SSOs). Results: We found out that HLA-B*46:01 (12.00%), HLA-B*15:02 (9.25%), HLA-B*40:01 (6.50%), HLA-B*13:01 (6.25%), and HLA-B* 38:02 (5.50%) alleles were more common than other alleles in Thai population. HLA-B*46:01 and HLA-B*15:02 were the most common allele found across four regions. Moreover, the frequency of HLA-B*15:01 and HLA-B*51:01 alleles were similarly distributed in Thai population (0.50, 5.25 %) and (p-value > 0.05), respectively. The frequencies of HLA-B*15:01 and HLA-B*51:01 alleles were not significantly different from other populations compared to the Thai population. Conclusions: We can screen for HLA-B*15:01 and HLA-B*51:01 alleles to determine the symptoms of COVID-19 since they are universal HLA-B markers. Importantly, the database of HLA markers indicates the association between HLA frequency and populations. However, we need further research on larger numbers of COVID-19 patients and in different populations.

Keywords: HLA-B*15:01, HLA-B*51:01, COVID-19, HLA-B alleles

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Authors: Jai Myung Yang, Na Young Kim, Sung Ho Shin

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The adverse reactions of current live smallpox vaccines and potential use of smallpox as a bioterror weapon have heightened the development of new effective vaccine for this infectious disease. In the present study, DNA vaccine vector was produced which was optimized for expression of the vaccinia virus L1 antigen in the mouse model. A plasmid IgM-tL1R, which contains codon-optimized L1R gene, was constructed and fused with an IgM signal sequence under the regulation of a SV40 enhancer. The expression and secretion of recombinant L1 protein was confirmed in vitro 293 T cell. Mice were administered the DNA vaccine by electroporation and challenged with vaccinia virus. We observed that immunization with IgM-tL1R induced potent neutralizing antibody responses and provided complete protection against lethal vaccinia virus challenge. Isotyping studies reveal that immunoglobulin G2 (IgG2) antibody predominated after the immunization, indicative of a T helper type 1 response. Our results suggest that an optimized DNA vaccine, IgM-tL1R, can be effective in stimulating anti-vaccinia virus immune response and provide protection against lethal orthopoxvirus challenge.

Keywords: DNA vaccine, electroporation, L1R, vaccinia virus

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Authors: Liu Yuhong, Hussen Mansai, Mei Chunli

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Inflammatory discitis is a sterile inflammatary disease that typically presents with abnormalities in two adjacent vertebral bodies and the intervening disk. Diagnosis this disorder is usually difficult and ideal management remains controversial. In this report，we examine a case of inflammatory discitis in a 56 year old female in which treatment with adalimumab ameliorated symptoms. The 56-year-old female patient developed repeatedly inflammatory discitis in the past three years, presenting with severe back pain, an elevated C-reactive protein and erythrocyte sedimentation rate, radiological erosive changes in vertebral and intervertebral disk of the spine. Surgical treatment, antibiotics and non steroidal anti-inflammatory drugs(NSAIDs) were used, but the patient still suffered from recurrent onset of unbearable backache. Three years later from the patient’s first admission，adalimumab was prescribed due to the third occurrence of Anderson lesions, which she had been suffering from for years. Soon after the same day of adalimumab therapy, her symptoms had a dramatic improvement. On the following day she could stand and walk slowly, her CRP and ESR were decreased to nearly normal levels in 4 weeks. Human leukocyte antigen (HLA)-typing analysis revealed a positive result for HLA-B27, the patient’s inflammatory discitis was considered to be associated with spondyloarthropathy.

Keywords: adalimumab, inflammatory discitis, spondyloarthropathy, patient

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Authors: Asma Alhuqail, Nagwa Aref

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

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

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Authors: Anna Gaspard, Stéphane Boudreau, Martin Simard

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Altered temperature and precipitation regimes associated with climate change generally result in improved conditions for plant growth. For Arctic and sub-Arctic ecosystems, this new climatic context favours an increase in primary productivity, a phenomenon often referred to as "greening". The development of an erect shrub cover has been identified as the main driver of Arctic greening. Although this phenomenon has been widely documented at the circumpolar scale, little information is available at the scale of plant communities, the basic unit of the Arctic, and sub-Arctic landscape mosaic. The objective of this study is to quantify the variation of NDVI within the different plant communities of Nunavik, which will allow us to identify the plant formations that contribute the most to the increase in productivity observed in this territory. To do so, the variation of NDVI extracted from Landsat images for the period 1984 to 2020 was quantified. From the Landsat scenes, annual summer NDVI mosaics with a resolution of 30 m were generated. The ecological mapping of Northern Quebec vegetation was then overlaid on the time series of NDVI maps to calculate the average NDVI per vegetation polygon for each year. Our results show that NDVI increases are more important for the bioclimatic domains of forest tundra and erect shrub tundra, and shrubby formations. Surface deposits, variations in mean annual temperature, and variations in winter precipitation are involved in NDVI variations. This study has thus allowed us to quantify changes in Nunavik's vegetation communities, using fine spatial resolution satellite imagery data.

Keywords: climate change, latitudinal gradient, plant communities, productivity

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Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

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Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA₄₄₀₄. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: tPA, recombinant, transgenic, tobacco

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Authors: Shahzad Bhatti, M. Aslamkhan, Sana Abbas, Marcella Attimonelli, Hikmet Hakan Aydin, Erica Martinha Silva de Souza,

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Despite being situated at the crossroad of Asia, Pakistan has gained crucial importance because of its pivotal role in subsequent migratory events. To highlight the genetic footprints and to contribute an enigmatic picture of the relative population expansion pattern among four major Pashtun tribes in Khyber Pakhtunkhwa viz., Bangash, Khattak, Mahsuds and Orakzai, the complete mitochondrial control region of 100 Pashtun were analyzed. All Pashtun tribes studied here revealed high genetic diversity; that was comparable to the other Central Asian, Southeast Asian and European populations. The configuration of genetic variation and heterogeneity further unveiled through Multidimensional Scaling, Principal Component Analysis, and phylogenetic analysis. The results revealed that the Pashtun is a composite mosaic of West Eurasian ancestry of numerous geographic origin. They received substantial gene flow during different invasions and have a high element of the Western provenance. The most common haplogroups reported in this study are: South Asian haplogroup M (28%) and R (8%); whereas, West Asians haplogroups are present, albeit in high frequencies (67%) and widespread over all; HV (15%), U (17%), H (9%), J (8%), K (8%), W (4%), N (3%) and T (3%). Herein we linked the unexplored genetic connection between Ashkenazi Jews and Pashtun. The presence of specific haplotypes J1b (4%) and K1a1b1a (5%) point to a genetic connection of Jewish conglomeration with Khattak tribe. This was a result of an ancient genetic influx in the early Neolithic period that led to the formation of a diverse genetic substratum in present day Pashtun.

Keywords: mtDNA haplogroups, control region, Pakistan, KPK, ethnicity

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Authors: Ashrafosadat Mousavi Laer, Elaheh Moravej

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The art of binding was simple and elementary at the beginning of Islam. This art thrived gradually and continued its development as an independent art. Identification of the binding techniques and used materials in covers and investigation of the arrays give us indexes for the better identification of different doctrines and methods of that time. The catalogers of the manuscripts usually pay attention to four items: gender, color, art elegances, injury, and exquisiteness of the cover. The criterion for classification of the covers is their art nature and gender. 15th century AD (9th century AH) was the period of the binding art development in which the most beautiful covers were produced by the so-called method of ‘burning’. At 16th century AD (10th century AH), in Safavid era, art changed completely and a fundamental evolution occurred in the technique and method of binding. The greatest change in this art was the extensive use of stamp that was made mostly of steel and copper. Theses stamps were presses against leather. These covers were called ‘beat’. In this paper, writing and bookbinding of about 32 Shahnamehs of Safavid era available in the Iranian libraries and museums are studied. An analytical-statistical study shows that four methods have been used including beat, burning, mosaic, and oily. 69 percent of the covers of these copies are cardboards with a leathery coating (goatskin) and have been produced by burning and beat methods. Its reasons are that these two methods have been common methods in Safavid era and performing them was only feasible on leather and the most desirable and commonly used leather of that time was goatskin which was the best option for cover legend durability and preserving the book and it was more durable because it had been made of goat skin. In addition, it had prepared a suitable opportunity for the binding artist’s creativity and innovation.

Keywords: Shahnameh, Safavid era, bookbinding, beat cover, burning cover

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Authors: Watchawin Ekphinitphithaya, Patompong Satapornpong

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Introduction: Carbamazepine (CBZ) is one of the most prescribed antiepileptic drugs (AEDs) by neurologists and non-neurologist worldwide. CBZ is usually prescribed along with other drugs, leading to the possibility of severe cutaneous adverse drug reactions (SCARs). The HLA-B*15:02 is strongly associated with CBZ-induced Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS–TEN) in the Han Chinese and other Asian populations but not in European populations, while HLA-A*31:01 allele has been reported to be associated with CBZ-induced SCARs in European population and Japanese. Objective: The aim of this study is to investigate the distribution of pharmacogenetics HLA-A*31:01 marker in a healthy Thai population associated with Carbamazepine-induced SCARs. Materials and Methods: Prospective study, 350 unrelated healthy Thais were recruited in this study. Human leukocyte antigen-A alleles were genotyped using PCR-sequence specific oligonucleotides (PCR-SSOs). Results: The frequency of HLA-A alleles were HLA-A*11:01 (190 alleles, 27.14%), HLA-A*24:02 (82 alleles, 11.71%), HLA-A*02:03 (80 alleles, 11.43%), HLA-A*33:03 (76 alleles, 10.86%), HLA-A*02:07 (58 alleles, 8.29%), HLA-A*02:01 (35 alleles, 5.00%), HLA-A*24:07 (29 alleles, 4.14%), HLA-A*02:06 – HLA-A*30:01 (15 alleles, 2.14%), and HLA-A*01:01 (14 alleles, 2.00%). Particularly, the number of HLA-A*31:01 alleles was 6 of 700 (0.86%) in the healthy Thai population. Many research presented varying distributions of HLA-A*31:01 in Asians, including 2% of Han Chinese, 9% of Japanese and 5% of Koreans. In addition, this allele was found approximately 2-5% in the Caucasian population. Conclusions: Thus, the pharmacogenetics database is vital to support in many populations, especially in Thais, for screening HLA-A*31:01 allele to avoid CBZ-induced SCARs before initiating treatments in each population.

Keywords: Carbamazepine, HLA-A*31:01, Thai population, pharmacogenetics

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Authors: M. Toghyani, A. Ghasemi, S. A. Tabeidian

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The present study was carried out to evaluate the effect of different levels of dietary seed and extract of Harmal (Peganum harmala L.) on immunity of broiler chicks. A total of 350 one-day old broiler chicks (Ross 308) were randomly allocated to five dietary treatments with four replicates pen of 14 birds each. Dietary treatments consisted of control, 1 and 2 g/kg Harmal seed in diet, 100 and 200 mg/L Harmal seed extract in water. Broilers received dietary treatments from 1 to 42 d. Two birds from each pen were randomly weighed and sacrificed at 42 d of age, the relative weight of lymphoid organs (bursa of Fabercius and spleen) to live weight were calculated. Antibody titers against Newcastle and influenza viruses and sheep red blood cell were measured at 30 d of age. Results showed that the relative weights of lymphoid organs were not affected by dietary treatments. Furthermore, antibody titer against Newcastle and influenza viruses as well as sheep red blood cell antigen were significantly (P<0.05) enhanced by feeding Harmal seed and extract. In conclusion, the results indicated that dietary inclusion of Harmal seed and extract enhanced immunological responses in broiler chicks.

Keywords: broiler chicks, Harmal, immunity, Peganum harmala

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Authors: Zainul A. Khan, Malik Z. Abdin, Jawaid A. Khan

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Begomoviruses belonging to the family Geminiviridae, have single-stranded circular DNA genomes that are monopartite or bipartite. The large intergenic region (LIR) of the monopartite and common region (CR) of bipartite begomoviruses possess promoter activity in their genomes. In this study, we have characterized novel bidirectional promoters from Cotton leaf curl Burewala virus (CLCuBuV) genome using high-throughput software and analyzed with PlantCARE, PLACE, Cister and PlantPAN databases. The promoters (Rep and CP promoters) were assayed both in stable and transient expression systems in tobacco as well as cotton plants. Rep and CP-based promoters from the LIR sequence of CLCuBuV and 35S promoter of Cauliflower mosaic virus (CaMV) were tagged with β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes to check the efficacy of the promoters. Histochemical staining of GUS in transformed tobacco (Nicotiana tabacum cv. Xanthi) leaves showed higher GUS expression driven by CLCuBuV Rep (complimentary sense) promoter as compared to conventional CaMV 35S promoter and CLCuBuV CP (virion sense) promoter, respectively. GUS activity in individual plant cells driven by CLCuBuV Rep, CLCuBuV CP, and CaMV 35S promoter were quantified through fluorometric GUS assay and reverse transcription quantitative real-time PCR (RT-qPCR). The expression level of GUS tagged with CLCuBuV Rep promoter in the transformed tobacco plants was obtained 2 to 4 fold higher than CaMV 35S promoter. When CLCuBuV CP promoter was used, lower expression level was monitored than that by CaMV 35S promoter. The expression of GFP-tagged with CLCuBuV promoters was also investigated through agroinfiltration. The CLCuBuV Rep promoters showed stronger consistent transient expression in the leaves of N. benthamiana, N. tabacum and Gossypium hirsutum plants when compared with CaMV 35S and CLCuBuV CP promoter.

Keywords: Begmovirus, bidirectional promoter, CaMV 35S promoter, GFP, GUS, qPCR

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Authors: Sawarni Hasibuan, Juliza Hidayati

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Competitiveness and sustainability issues not only put pressure on big companies, but also small and medium enterprises (SMEs). SMEs Batik Bogor is one of the local culture-based creative industries in Bogor city which is also dealing with the issue of sustainability. The purpose of this research is to develop framework of sustainability at SMEs Batik Indonesia case of SMEs Batik Bogor by integrating innovation of cleaner production in its supply chain. The approach used is desk study, field survey, in-depth interviews, and benchmarking best practices of SMEs sustainability. In-depth interviews involve stakeholders to identify the needs and standards of sustainability of SMEs Batik. Data analysis was done by benchmarking method, Multi Dimension Scaling (MDS) method, and Strength, Weakness, Opportunity, Threat (SWOT) analysis. The results recommend the framework of sustainability for SMEs Batik in Indonesia. The sustainability status of SMEs Batik Bogor is classified as Moderate Sustainable. Factors that support the sustainability of SMEs Batik Bogor such is a strong commitment of top management in adopting cleaner production innovation and creativity approach. Successful cleaner production innovations are implemented primarily in the substitution of dye materials from toxic to non-toxic, reducing the intensity of non-renewable energy use, as well as the reuse and recycle of solid waste. “Mosaic Batik” is one of the innovations of solid waste utilization of batik waste produced by company R&D center that gives benefit to three pillars of sustainability, that is financial benefit, environmental benefit, and social benefit. The sustainability of SMEs Batik Bogor cannot be separated from the support of Bogor City Government which proactively facilitates the promotion of sustainable innovation produced by SMEs Batik Bogor.

Keywords: cleaner production innovation, creativity, SMEs Batik, sustainability supply chain

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Authors: Dharmendra Pratap

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Large Cardamom (Amomum subulatum), family Zingiberaceae is an aromatic spice crop and has rich medicinal value. Large Cardamom is as synonymous to Sikkim as Tea is to Darjeeling. Since Sikkim alone contributes up to 88% of India's large cardamom production which is the world leader by producing over 50% of the global yield. However, the production of large cardamom has declined almost to half since last two decade. The economic losses have been attributed to two viral diseases namely, chirke and Foorkey. Chirke disease is characterized by light and dark green streaks on leaves. The affected leaves exhibit streak mosaic, which gradually coalesce, turn brown and eventually dry up. Excessive sprouting and formation of bushy dwarf clumps at the base of mother plants that gradually die characterize the foorkey disease. In our surveys in Sikkim–Darjeeling hill area during 2012-14, 40-45% of plants were found to be affected with foorkey disease and 10-15% with chirke. Mechanical and aphid transmission study showed banana as an alternate host for both the disease. For molecular identification, total genomic DNA and RNA was isolated from the infected leaf tissues and subjected to Rolling circle amplification (RCA) and RT-PCR respectively. The DNA concatamers produced in the RCA reaction were monomerized by different restriction enzymes and the bands corresponding to ~1 kb genomes were purified and cloned in the respective sites. The nucleotide sequencing results revealed the association of Nanovirus with the foorkey disease of large cardamom. DNA1 showed 74% identity with Replicase gene of FBNYV, DNA2 showed 77% identity with the NSP gene of BBTV and DNA3 showed 74% identity with CP gene of BBTV. The finding suggests the presence of a new species of nanovirus associated with foorkey disease of large cardamom in Sikkim and Darjeeling hills. The details of their epidemiology and other factors would be discussed.

Keywords: RCA, nanovirus, large cardamom, molecular virology and microbiology

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Authors: Aminu Abdullahi Isyaku

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Semi-arid Bornu basin in northeast Nigeria is characterised with flat topography, thick cover sediments and lack of continuous bedrock outcrops discernible for field geology. This paper presents the methodology for the characterisation of neotectonic surface structures and surface lithology in the north-eastern Bornu basin in northeast Nigeria as an alternative approach to field geological mapping using free multispectral Landsat 7 ETM+, SRTM DEM and ASAR Earth Observation datasets. Spectral lithological mapping herein developed utilised spectral discrimination of the surface features identified on Landsat 7 ETM+ images to infer on the lithology using four steps including; computations of band combination images; band ratio images; supervised image classification and inferences of the lithological compositions. Two complementary approaches to lineament mapping are carried out in this study involving manual digitization and automatic lineament extraction to validate the structural lineaments extracted from the Landsat 7 ETM+ image mosaic covering the study. A comparison between the mapped surface lineaments and lineament zones show good geospatial correlation and identified the predominant NE-SW and NW-SE structural trends in the basin. Topographic profiles across different parts of the Bama Beach Ridge palaeoshorelines in the basin appear to show different elevations across the feature. It is determined that most of the drainage systems in the northeastern Bornu basin are structurally controlled with drainage lines terminating against the paleo-lake border and emptying into the Lake Chad mainly arising from the extensive topographic high-stand Bama Beach Ridge palaeoshoreline.

Keywords: Bornu Basin, lineaments, spectral lithology, tectonics

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Authors: Puthsodary Tat

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Democracy has been categorially accepted and used as foreign and domestic policy agendas for the hope of peace, economic growth and prosperity for more than 25 years in Cambodia. However, the country is now in the grip of dictatorship, human rights violations, and prospective economic sanctions. This paper examines different perceptions and experiences of democratic assistance. In this study, the author employs discourse theory, idealism and realism as a theory-based methodology for debating and assessing the implications of democratization. Discourse theory is used to establish a platform for understanding discursive formations, body of knowledge and the games of truth of democracy. Idealist approaches give rational arguments for adopting key tenets that work well on the ground. In contrast, realism allows for some sweeping critiques of utopian ideal and offers particular views on why Western hegemonic missions do not work well. From idealist views, the research finds that Cambodian people still believe that democracy is a prima facie universality for peace, growth and prosperity. From realism, democratization is on the brink of death in three reasons. Firstly, there are tensions between Western and local discourses about democratic values and norms. Secondly, democratic tenets have been undermined by the ruling party-controlled courts, corruption, structural oppression and political patronage-based institutions. The third pitfall is partly associated with foreign aid dependency and geopolitical power struggles in the region. Finally, the study offers a precise mosaic of democratic principles that may be used to avoid a future geopolitical and economic crisis.

Keywords: corruption, democracy, democratic principles, discourse theory, discursive formations, foreign aid dependency, games of truth, geopolitical and economic crisis, geopolitical power struggle, hegemonic mission, idealism, realism, utopian ideal

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Authors: Saekil Yun, Sib Sankar Giri, Jin Woo Jun, Hyoun Joong Kim, Sang Guen Kim, Sang Wha Kim, Jung Woo Kang, Se Jin Han, Se Chang Park

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In aquaculture, vaccination is important to control and prevent diseases. In the study, we utilized poly D,L-lactide-co-glycolic acid (PLGA) microparticles (MPs) for encapsulating formalin-killed Aeromonas hydrophila cells. To assess the innate and adaptive immune responses, carps and loaches were used for the experiments. Fish were divided into three groups (A, B, C). Total antigen of 0.1 ml vaccine was adjusted by 2 x 108 CFU and injected via intraperitoneal route. Group A was vaccinated with 0.1 ml of PLGA vaccine, group B was with 0.1 ml of FKC vaccine and group C was with 0.1 ml of sterile PBS. All three groups were challenged with A. hydrophila and challenge dose was lethal dose (LD50). Loaches and carp were then challenged with A. hydrophila at 12 and 20 weeks post vaccination (wpv), and 10 and 14 wpv, respectively, and relative survival rates were calculated. For both fish species, the curve of antibody titer over time was shallower in the PLGA group than the FKC group and the PLGA groups demonstrated higher survival rates at all time-points. In the groups of PLGA-MP, relative mRNA levels of IL-1β, TNF-α, lysozyme C and IgM were significantly upregulated than FKC treated groups. Biodegradable PLGA microparticle vaccine could induce longer immune responses than original FKC vaccines to protect from A. hydrophila infection.

Keywords: PLGA, microparticles, Aeromonas hydrophila, vaccine

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Authors: Siriniya Siribrahmanakul

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Introduction:SARsCOVID-19 is rapidly spreading, and some people may exhibit severe symptoms. Mortality rate of 2.0–3.0% with COVID-19 infection atworldwide. Human leukocyte antigen (HLA), located on chromosome 6, consist of HLA class I and class II. HLA are used by the immune system to attach self-antigens. Previous studies, HLA-DRB1*01:01,HLA-DRB1*12:01and HLA-DRB1*14:04 were, showed significant difference with severe COVID-19 in the Chinese population by p-value < 0.05. Objective: We investigated the prevalence of HLA-DRB1 alleles associated with severe COVID-19 in Thai population. Materials and Methods:200 DNA samples were isolated from EDTA blood using the MagNAprue Compact Nucleic Acid Isolation kits.HLA-DRB1alleles were genotyped using sequence-specific oligonucleotides (PCR-SSOs). Results:The frequency of HLA-DRB1 alleles in Thai population wereHLA-DRB1*12:02 (15.75%), HLA-DRB1*15:02 (14.50%), HLA-DRB1*09:01 (11.50%), HLA-DRB1*07:01 (9.50%), HLA-DRB1*03:01,HLA-DRB1*05:01 (5.75%), HLA-DRB1*14:01 (5.50%), HLA-DRB1*16:02 (4.50%), HLA-DRB1*04:05 (4.00%), HLA-DRB1*14:03 (3.25%), HLA-DRB1*10:01 (2.25%) and HLA-DRB1*13:02 (2.00%). Particularly, HLA-DRB1*12:02 allele was the highest allele frequency presented in the four regions groups of Thai population. Furthermore, the HLA-DRB1* alleles associated with severe COVID-19, which consists ofHLA-DRB1*14:04(2.00 %) and HLA-DRB1*12:01(0.50%) in Thai population, whereas HLA-DRB1*01:01 allele was not found in this population. HLA-DRB1*14:04 and HLA-DRB1*12:01alleles were similarly distributed in four regions populations in Thailand (p-value > 0.05). The alleles frequencies of HLA-DRB1*14:04 and HLA-DRB1*12:01, which associated with severe COVID-19, had no significant differences between Thai population, South China population, South Africa population, and South Koreapopulation (p-value > 0.05). Conclusions: Particularly, this study has focused on allele frequency of HLA-DRB1*14:04in a healthy Thai population to evaluating their impact on the severe COVID-19. Furthermore, our research needs to be done in larger numbers of Thai patients.

Keywords: HLA-DRB1, allele frequency, Thai population, COVID-19 marker

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