MyMedR

Displaying publications 1 - 20 of 52 in total

Abstract:

Sort:

First Report of Spiroplasma citri (-Induced) Associated with Periwinkle Lethal Yellows in Southeast Asia

Nejat N, Vadamalai G, Sijam K, Dickinson M

Plant Dis, 2011 Oct;95(10):1312.
PMID: 30731679 DOI: 10.1094/PDIS-03-11-0251

Madagascar periwinkle, Catharanthus roseus (L.) G. Don, is a member of the Apocynaceae plant family that is native to Madagascar and produces dimeric terpenoid indole alkaloids that are used in the treatment of hypertension and cancer. Periwinkle as an indicator plant is highly susceptible to phytoplasmas and spiroplasma infection from different crops, and has been found to be naturally infected with spiroplasmas in Arizona, California, and the Mediterranean countries. In this study, surveys of suspected diseased periwinkles were conducted in various regions of Selangor State, Malaysia. Periwinkles showing rapid decline in the number and size of the flowers, premature abscission of buds and flowers, reduction in leaf size, chlorosis of the leaf tips and margins, general chlorosis, and stunting and dying plants were collected. These symptoms were widespread on periwinkle in this state. Diagnosis of the disease was based on symptomatology, grafting, serology (ELISA), PCR techniques, and cultivation. Tests for transmission by grafting were conducted using symptomatic periwinkle plants. Symptoms were induced on all eight graft-inoculated healthy periwinkles approximately 2 weeks after side grafting. Preliminary examination was performed by ELISA with Spiroplasma citri Saglio polyclonal antibody that was prepared against an Iranian S. citri isolate (H. Rahimian, unpublished data). Leaf extracts of all 24 symptomatic periwinkles gave positive ELISA reactions at OD405 readings ranging from 0.310 to 0.654 to the antibody against S. citri by the indirect ELISA method. Six healthy periwinkle leaves gave OD405 readings around 0.128. Total nucleic acids were extracted from 10 symptomatic and 5 asymptomatic plants (4). PCR using the ScR16F1/ScR16R1 primer pair designed to detect S. citri in carrot and P1/P7 and secA for1/rev3 primer pairs designed for identification of phytoplasmas were used to detect the causal agent (1-3). Amplification failed when the P1/P7 universal phytoplasma primer pair was used for diseased samples. However, the PCR assays resulted in products of 1,833 and 800 bp with ScR16F1/ScR16R1 and secA for1/rev3, respectively. Five of each ScR16F1/ScR16R1 and SecAfor1/SecArev3 products were cloned with the Topo TA cloning kit (Invitrogen, Carlsbad, CA), sequenced, and deposited as GenBank Accession Nos. HM015669 and FJ011099, respectively. Sequences for both genes indicated that S. citri was associated with the disease on periwinkle. ScR16F1/ScR16R1 products cloned from symptomatic periwinkles had 98% sequence identity with S. citri (GenBank Accession No. AM285316), while nucleotide sequences of SecAfor1/SecArev3 products had 88% sequence identity with S. citri GII3-3X (GenBank Accession No. AM285304). S. citri was cultivated from 10 S. citri-infected periwinkles using filtration and SP-4 media. Twenty culture tubes started to change culture medium color from red to yellow 1 month after cultivation. Helical and motile S. citri was observed in the dark-field microscope. To our knowledge, this is the first report on the presence and occurrence of S. citri in Southeast Asia and its association with lethal yellows on periwinkle in Malaysia. References: (1) J. Hodgetts et al. Int. J. Syst. Evol. Microbiol. 58:1826, 2008. (2) I.-M. Lee et al. Phytopathology 85:728, 1995. (3) I.-M. Lee et al. Plant Dis. 90:989, 2006. (4) Y.-P. Zhang et al. J. Virol. Methods. 71:45, 1998.

Matched MeSH terms: Nucleic Acids
Fulltext Transcriptomic profiling of skeletal muscle from the Ts1Cje mouse model of Down syndrome suggests dysregulation of trisomic genes associated with neuromuscular junction signaling, oxidative stress and chronic inflammation

Leong, Melody Pui Yee, Usman Bala, Lim, Chai Ling, Rozita Rosli, Cheah, Pike-See, Ling, King-Hwa

Neuroscience Research Notes, 2018;1(1):21-41.
MyJurnal

Ts1Cje is a mouse model of Down syndrome (DS) with partial triplication of chromosome 16, which encompasses a high number of human chromosome 21 (HSA21) orthologous genes. The mouse model exhibits muscle weakness resembling hypotonia in DS individuals. The effect of extra gene dosages on muscle weakness or hypotonia in Ts1Cje and DS individuals remains unknown. To identify molecular dysregulation of the skeletal muscle, we compared the transcriptomic signatures of soleus and extensor digitorum longus (EDL) muscles between the adult Ts1Cje and disomic littermates. A total of 166 and 262 differentially expressed protein-coding genes (DEGs) were identified in the soleus and EDL muscles, respectively. The partial trisomy of MMU16 in Ts1Cje mice has a greater effect on gene expression in EDL. Top-down clustering analysis of all DEGs for represented functional ontologies revealed 5 functional clusters in soleus associated with signal transduction, development of reproductive system, nucleic acid biosynthesis, protein modification and metabolism as well as regulation of gene expression. On the other hand, only 3 functional clusters were observed for EDL namely neuron and cell development, protein modification and metabolic processes as well as ion transport. A total of 11 selected DEGs were validated using qPCR (disomic DEGs: Mansc1; trisomic DEGs: Itsn1, Rcan1, Synj1, Donson, Dyrk1a, Ifnar1, Ifnar2, Runx1, Sod1 and Tmem50b). The validated DEGs were implicated in neuromuscular junction signalling (Itsn1, Syn1), oxidative stress (Sod1, Runx1) and chronic inflammation processes (Runx1, Rcan1, Ifnar1, Ifnar2). Other validated DEGs have not been well-documented as involved in the skeletal muscle development or function, thus serve as interesting novel candidates for future investigations. To our knowledge, the study was the first attempt to determine the transcriptomic profiles of both soleus and EDL muscles in Ts1Cje mice. It provides new insights on the possible disrupted molecular pathways associated with hypotonia in DS individuals.

Matched MeSH terms: Nucleic Acids
The Occurrence of Bean common mosaic virus and Cucumber mosaic virus in Yardlong Beans in Indonesia

Damayanti TA, Alabi OJ, Rauf A, Naidu RA

Plant Dis, 2010 Apr;94(4):478.
PMID: 30754487 DOI: 10.1094/PDIS-94-4-0478B

Yardlong bean (Vigna unguiculata subsp. sesquipedalis) is extensively cultivated in Indonesia for consumption as a green vegetable. During the 2008 season, a severe outbreak of a virus-like disease occurred in yardlong beans grown in farmers' fields in Bogor, Bekasi, Subang, Indramayu, and Cirebon of West Java, Tanggerang of Banten, and Pekalongan and Muntilan of Central Java. Leaves of infected plants showed severe mosaic to bright yellow mosaic and vein-clearing symptoms, and pods were deformed and also showed mosaic symptoms on the surface. In cv. 777, vein-clearing was observed, resulting in a netting pattern on symptomatic leaves followed by death of the plants as the season advanced. Disease incidence in the Bogor region was approximately 80%, resulting in 100% yield loss. Symptomatic leaf samples from five representative plants tested positive in antigen-coated plate-ELISA with potyvirus group-specific antibodies (AS-573/1; DSMZ, German Resource Center for Biological Material, Braunschweig, Germany) and antibodies to Cucumber mosaic virus (CMV; AS-0929). To confirm these results, viral nucleic acids eluted from FTA classic cards (FTA Classic Card, Whatman International Ltd., Maidstone, UK) were subjected to reverse transcription (RT)-PCR using potyvirus degenerate primers (CIFor: 5'-GGIVVIGTIGGIWSIGGIAARTCIAC-3' and CIRev: 5'-ACICCRTTYTCDATDATRTTIGTIGC-3') (3) and degenerate primers (CMV-1F: 5'-ACCGCGGGTCTTATTATGGT-3' and CMV-1R: 5' ACGGATTCAAACTGGGAGCA-3') specific for CMV subgroup I (1). A single DNA product of approximately 683 base pairs (bp) with the potyvirus-specific primers and a 382-bp fragment with the CMV-specific primers were amplified from ELISA-positive samples. These results indicated the presence of a potyvirus and CMV as mixed infections in all five samples. The amplified fragments specific to potyvirus (four samples) and CMV (three samples) were cloned separately into pCR2.1 (Invitrogen Corp., Carlsbad, CA). Two independent clones per amplicon were sequenced from both orientations. Pairwise comparison of these sequences showed 93 to 100% identity among the cloned amplicons produced using the potyvirus-specific primers (GenBank Accessions Nos. FJ653916, FJ653917, FJ653918, FJ653919, FJ653920, FJ653921, FJ653922, FJ653923, FJ653924, FJ653925, and FJ653926) and 92 to 97% with a corresponding nucleotide sequence of Bean common mosaic virus (BCMV) from Taiwan (No. AY575773) and 88 to 90% with BCMV sequences from China (No. AJ312438) and the United States (No. AY863025). The sequence analysis indicated that BCMV isolates from yardlong bean are more closely related to an isolate from Taiwan than with isolates from China and the United States. The CMV isolates (GenBank No. FJ687054) each were 100% identical and 96% identical with corresponding sequences of CMV subgroup I isolates from Thailand (No. AJ810264) and Malaysia (No. DQ195082). Both BCMV and CMV have been documented in soybean, mungbean, and peanut in East Java of Indonesia (2). Previously, BCMV, but not CMV, was documented on yardlong beans in Guam (4). To our knowledge, this study represents the first confirmed report of CMV in yardlong bean in Indonesia and is further evidence that BCMV is becoming established in Indonesia. References: (1) J. Aramburu et al. J. Phytopathol. 155:513, 2007. (2) S. K. Green et al. Plant Dis. 72:994, 1988. (3) C. Ha et al. Arch. Virol. 153:25, 2008. (4) G. C. Wall et al. Micronesica 29:101, 1996.

Matched MeSH terms: Nucleic Acids
Zeolite-iron oxide nanocomposite from fly ash formed a 'clubbell' structure: integration of cardiac biocapture macromolecules in serum on microelectrodes

Liu Z, Gopinath SCB, Wang Z, Li Y, Anbu P, Zhang W

Mikrochim Acta, 2021 05 15;188(6):187.
PMID: 33990848 DOI: 10.1007/s00604-021-04834-w

A new zeolite-iron oxide nanocomposite (ZEO-IO) was extracted from waste fly ash of a thermal power plant and utilized for capturing aptamers used to quantify the myocardial infarction (MI) biomarker N-terminal prohormone B-type natriuretic peptide (NT-ProBNP); this was used in a probe with an integrated microelectrode sensor. High-resolution microscopy revealed that ZEO-IO displayed a clubbell structure and a particle size range of 100-200 nm. Energy-dispersive X-ray spectroscopy and X-ray photoelectron spectroscopy confirmed the presence of Si, Al, Fe, and O in the synthesized ZEO-IO. The limit of detection for NT-ProBNP was 1-2 pg/mL (0.1-0.2 pM) when the aptamer was sandwiched with antibody and showed the doubled current response even at a low NT-ProBNP abundance. A dose-dependent interaction was identified for this sandwich with a linear plot in the concentration range 1 to 32 pg/mL (0.1-3.2 pM) with a determination coefficient R2 = 0.9884; y = 0.8425x-0.5771. Without sandwich, the detection limit was 2-4 pg/mL (0.2-0.4 pM) and the determination coefficient was R2 = 0.9854; y = 1.0996x-1.4729. Stability and nonfouling assays in the presence of bovine serum albumin, cardiac troponin I, and myoglobin revealed that the aptamer-modified surface is stable and specific for NT-Pro-BNP. Moreover, NT-ProBNP-spiked human serum exhibited selective detection. This new nanocomposite-modified surface helps in detecting NT-Pro-BNP and diagnosing MI at stages of low expression.

Matched MeSH terms: Immobilized Nucleic Acids/chemistry
Development of a dry-reagent-based nucleic acid-sensing platform by coupling thermostabilised LATE-PCR assay to an oligonucleotide-modified lateral flow biosensor

Ang GY, Yu CY, Chan KG, Singh KK, Chan Yean Y

J Microbiol Methods, 2015 Nov;118:99-105.
PMID: 26342435 DOI: 10.1016/j.mimet.2015.08.024

In this study, we report for the first time the development of a dry-reagent-based nucleic acid-sensing platform by combining a thermostabilised linear-after-the-exponential (LATE)-PCR assay with a one-step, hybridisation-based nucleic acid lateral flow biosensor. The nucleic acid-sensing platform was designed to overcome the need for stringent temperature control during transportation or storage of reagents and reduces the dependency on skilled personnel by decreasing the overall assay complexity and hands-on time. The platform was developed using toxigenic Vibrio cholerae as the model organism due to the bacterium's propensity to cause epidemic and pandemic cholera. The biosensor generates result which can be visualised with the naked eyes and the limit of detection was found to be 1pg of pure genomic DNA and 10CFU/ml of toxigenic V. cholerae. The dry-reagent-based nucleic acid-sensing platform was challenged with 95 toxigenic V. cholerae, 7 non-toxigenic V. cholerae and 66 other bacterial strains in spiked stool sample and complete agreement was observed when the results were compared to that of monosialoganglioside (GM1)-ELISA. Heat-stability of the thermostabilised LATE-PCR reaction mixes at different storage temperatures (4-56°C) was investigated for up to 90days. The dry-reagent-based genosensing platform with ready-to-use assay components provides an alternative method for sequence-specific detection of nucleic acid without any cold chain restriction that is associated with conventional molecular amplification techniques.

Matched MeSH terms: Nucleic Acids
Fulltext Advances in the Diagnosis of Foot-and-Mouth Disease

Wong CL, Yong CY, Ong HK, Ho KL, Tan WS

Front Vet Sci, 2020;7:477.
PMID: 32974392 DOI: 10.3389/fvets.2020.00477

Foot-and-mouth disease (FMD) is a devastating livestock disease caused by foot-and-mouth disease virus (FMDV). Outbreaks of this disease in a country always result in conspicuous economic losses to livestock industry and subsequently lead to serious socioeconomic damages due to the immediate imposition of trade embargo. Rapid and accurate diagnoses are imperative to control this infectious virus. In the current review, enzyme-linked immunosorbent assay (ELISA)-based methods used in FMD diagnosis are extensively reviewed, particularly the sandwich, liquid-phase blocking, and solid-phase competition ELISA. The differentiation of infected animals from vaccinated animals using ELISA-based methods is also highlighted, in which the role of 3ABC polyprotein as a marker is reviewed intensively. Recently, more studies are focusing on the molecular diagnostic methods, which detect the viral nucleic acids based on reverse transcription-polymerase chain reaction (RT-PCR) and RT-loop-mediated isothermal amplification (RT-LAMP). These methods are generally more sensitive because of their ability to amplify a minute amount of the viral nucleic acids. In this digital era, the RT-PCR and RT-LAMP are progressing toward the mobile versions, aiming for on-site FMDV diagnosis. Apart from RT-PCR and RT-LAMP, another diagnostic assay specifically designed for on-site diagnosis is the lateral flow immunochromatographic test strips. These test strips have some distinct advantages over other diagnostic methods, whereby the assay often does not require the aid of an external device, which greatly lowers the cost per test. In addition, the on-site diagnostic test can be easily performed by untrained personnel including farmers, and the results can be obtained in a few minutes. Lastly, the use of FMDV diagnostic assays for progressive control of the disease is also discussed critically.

Matched MeSH terms: Nucleic Acids
Fulltext Combinatorial Antimicrobial Efficacy and Mechanism of Linalool Against Clinically Relevant Klebsiella pneumoniae

Yang SK, Yusoff K, Ajat M, Wee CY, Yap PS, Lim SH, et al.

Front Microbiol, 2021;12:635016.
PMID: 33815320 DOI: 10.3389/fmicb.2021.635016

Antibiotic-adjuvant combinatory therapy serves as a viable treatment option in addressing antibiotic resistance in the clinical setting. This study was carried out to assess and characterize the adjuvant potential and mode of action of linalool against carbapenemase-producing Klebsiella pneumoniae (KPC-KP). Linalool exhibited bactericidal activity alone (11,250 μg/ml) and in combination with meropenem (5,625 μg/ml). Comparative proteomic analysis showed significant reduction in the number of cytoplasmic and membrane proteins, indicating membrane damage in linalool-treated KPC-KP cells. Upregulation of oxidative stress regulator proteins and downregulation of oxidative stress-sensitive proteins indicated oxidative stress. Zeta potential measurement and outer membrane permeability assay revealed that linalool increases the bacterial surface charge as well as the membrane permeability. Intracellular leakage of nucleic acid and proteins was detected upon linalool treatment. Scanning and transmission electron microscopies further revealed the breakage of bacterial membrane and loss of intracellular materials. Linalool induced oxidative stress by generating reactive oxygen species (ROS) which initiates lipid peroxidation, leading to damage of the bacterial membrane. This leads to intracellular leakage, eventually killing the KPC-KP cells. Our study demonstrated that linalool possesses great potential in future clinical applications as an adjuvant along with existing antibiotics attributed to their ability in disrupting the bacterial membrane by inducing oxidative stress. This facilitates the uptake of antibiotics into the bacterial cells, enhancing bacterial killing.

Matched MeSH terms: Nucleic Acids
Fulltext Comparison of neurotoxic effects of ethanol and endosulfan on biochemical changes of brain tissues in Javanese medaka (Oryzias Javanicus) and zebrafish (Danio Reri

Nurul Farhana Ramlan, Noraini Abu Bakar, Albert, Emmellie Laura, Syaizwan Zahmir Zulkifli, Syahida Ahmad, Mohammad Noor Amal Azmai, et al.

Pertanika Journal of Science & Technology, 2020;28(2):689-701.
MyJurnal

An ideal model organism for neurotoxicology research should meet several characteristics, such as low cost and amenable for high throughput testing. Javanese medaka (JM) has been widely used in the ecotoxicological studies related to the marine and freshwater environment, but rarely utilized for biomedical research. Therefore, in this study, the applicability of using JM in the neurotoxicology research was assessed using biochemical comparison with an established model organism, the zebrafish. Identification of biochemical changes due to the neurotoxic effects of ethanol and endosulfan was assessed using Fourier Transform Infrared (FTIR) analysis. Treatment with ethanol affected the level of lipids, proteins, glycogens and nucleic acids in the brain of JM. Meanwhile, treatment with endosulfan showed alteration in the level of lipids and nucleic acids. For the zebrafish, exposure to ethanol affected the level of protein, fatty acid and amino acid, and exposure to endosulfan induced alteration in the fatty acids, amino acids, nucleic acids and protein in the brain of zebrafish. The sensitive response of the JM toward chemicals exposure proved that it was a valuable model for neurotoxicology research. More studies need to be conducted to further develop JM as an ideal model organism for neurotoxicology research.

Matched MeSH terms: Nucleic Acids
Fulltext Nucleic Acid-Based Diagnostic Tests for the Detection SARS-CoV-2: An Update

Yu CY, Chan KG, Yean CY, Ang GY

Diagnostics (Basel), 2021 Jan 01;11(1).
PMID: 33401392 DOI: 10.3390/diagnostics11010053

The coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) began as a cluster of pneumonia cases in Wuhan, China before spreading to over 200 countries and territories on six continents in less than six months. Despite rigorous global containment and quarantine efforts to limit the transmission of the virus, COVID-19 cases and deaths have continued to increase, leaving devastating impacts on the lives of many with far-reaching effects on the global society, economy and healthcare system. With over 43 million cases and 1.1 million deaths recorded worldwide, accurate and rapid diagnosis continues to be a cornerstone of pandemic control. In this review, we aim to present an objective overview of the latest nucleic acid-based diagnostic tests for the detection of SARS-CoV-2 that have been authorized by the Food and Drug Administration (FDA) under emergency use authorization (EUA) as of 31 October 2020. We systematically summarize and compare the principles, technologies, protocols and performance characteristics of amplification- and sequencing-based tests that have become alternatives to the CDC 2019-nCoV Real-Time RT-PCR Diagnostic Panel. We highlight the notable features of the tests including authorized settings, along with the advantages and disadvantages of the tests. We conclude with a brief discussion on the current challenges and future perspectives of COVID-19 diagnostics.

Matched MeSH terms: Nucleic Acids
Fulltext Antimicrobial activity and mode of action of terpene linalyl anthranilate against carbapenemase-producing Klebsiella pneumoniae

Yang SK, Yusoff K, Ajat M, Yap WS, Lim SE, Lai KS

J Pharm Anal, 2021 Apr;11(2):210-219.
PMID: 34012697 DOI: 10.1016/j.jpha.2020.05.014

Mining of plant-derived antimicrobials is the major focus at current to counter antibiotic resistance. This study was conducted to characterize the antimicrobial activity and mode of action of linalyl anthranilate (LNA) against carbapenemase-producing Klebsiella pneumoniae (KPC-KP). LNA alone exhibited bactericidal activity at 2.5% (V/V), and in combination with meropenem (MPM) at 1.25% (V/V). Comparative proteomic analysis showed a significant reduction in the number of cytoplasmic and membrane proteins, indicating membrane damage in LNA-treated KPC-KP cells. Up-regulation of oxidative stress regulator proteins and down-regulation of oxidative stress-sensitive proteins indicated oxidative stress. Zeta potential measurement and outer membrane permeability assay revealed that LNA increases both bacterial surface charge and membrane permeability. Ethidium bromide influx/efflux assay showed increased uptake of ethidium bromide in LNA-treated cells, inferring membrane damage. Furthermore, intracellular leakage of nucleic acid and proteins was detected upon LNA treatment. Scanning and transmission electron microscopies again revealed the breakage of bacterial membrane and loss of intracellular materials. LNA was found to induce oxidative stress by generating reactive oxygen species (ROS) that initiate lipid peroxidation and damage the bacterial membrane. In conclusion, LNA generates ROS, initiates lipid peroxidation, and damages the bacterial membrane, resulting in intracellular leakage and eventually killing the KPC-KP cells.

Matched MeSH terms: Nucleic Acids
Fulltext PH-responsive strontium nanoparticles for targeted gene therapy against mammary carcinoma cells

Bakhtiar A, Chowdhury EH

Asian J Pharm Sci, 2021 Mar;16(2):236-252.
PMID: 33995617 DOI: 10.1016/j.ajps.2020.11.002

Genetic intervention via the delivery of functional genes such as plasmid DNA (pDNA) and short-interfering RNA (siRNA) offers a great way to treat many single or multiple genetic defects effectively, including mammary carcinoma. Delivery of naked therapeutic genes or siRNAs is, however, short-lived due to biological clearance by scavenging nucleases and circulating monocytes. Low cellular internalization of negatively-charged nucleic acids further causes low transfection or silencing activity. Development of safe and effectual gene vectors is therefore undeniably crucial to the success of nucleic acid delivery. Inorganic nanoparticles have attracted considerable attention in the recent years due to their high loading capacity and encapsulation activity. Here we introduce strontium salt-based nanoparticles, namely, strontium sulfate, strontium sulfite and strontium fluoride as new inorganic nanocarriers. Generated strontium salt particles were found to be nanosized with high affinity towards negatively-charged pDNA and siRNA. Degradation of the particles was seen with a drop in pH, suggesting their capacity to respond to pH change and undergo dissolution at endosomal pH to release the genetic materials. While the particles are relatively nontoxic towards the cells, siRNA-loaded SrF2 and SrSO3 particles exerted superior transgene expression and knockdown activity of MAPK and AKT, leading to inhibition of their phosphorylation to a distinctive extent in both MCF-7 and 4T1 cells. Strontium salt nanoparticles have thus emerged as a promising tool for applications in cancer gene therapy.

Matched MeSH terms: Nucleic Acids
Fulltext Glutathione (GSH) improves sperm quality and testicular morphology in streptozotocin-induced diabetic mice

Abdullah F, Khan Nor-Ashikin MN, Agarwal R, Kamsani YS, Abd Malek M, Bakar NS, et al.

Asian J Androl, 2021 1 22;23(3):281-287.
PMID: 33473013 DOI: 10.4103/aja.aja_81_20

Diabetes mellitus (DM) is known to cause reproductive impairment. In men, it has been linked to altered sperm quality and testicular damage. Oxidative stress (OS) plays a pivotal role in the development of DM complications. Glutathione (GSH) is a part of a nonenzymatic antioxidant defense system that protects lipid, protein, and nucleic acids from oxidative damage. However, the protective effects of exogenous GSH on the male reproductive system have not been comprehensively examined. This study determined the impact of GSH supplementation in ameliorating the adverse effect of type 1 DM on sperm quality and the seminiferous tubules of diabetic C57BL/6NTac mice. GSH at the doses of 15 mg kg-1 and 30 mg kg-1 was given intraperitoneally to mice weekly for 6 consecutive weeks. The mice were then weighed, euthanized, and had their reproductive organs excised. The diabetic (D Group) showed significant impairment of sperm quality and testicular histology compared with the nondiabetic (ND Group). Diameters of the seminiferous lumen in diabetic mice treated with 15 mg kg-1 GSH (DGSH15) were decreased compared with the D Group. Sperm motility was also significantly increased in the DGSH15 Group. Improvement in testicular morphology might be an early indication of the protective roles played by the exogenous GSH in protecting sperm quality from effects of untreated type 1 DM or diabetic complications. Further investigation using different doses and different routes of GSH is necessary to confirm this suggestion.

Matched MeSH terms: Nucleic Acids
Fulltext Loop-mediated isothermal amplification (LAMP) reaction as viable PCR substitute for diagnostic applications: a comparative analysis study of LAMP, conventional PCR, nested PCR (nPCR) and real-time PCR (qPCR) based on Entamoeba histolytica DNA derived from faecal sample

Foo PC, Nurul Najian AB, Muhamad NA, Ahamad M, Mohamed M, Yean Yean C, et al.

BMC Biotechnol, 2020 Jun 22;20(1):34.
PMID: 32571286 DOI: 10.1186/s12896-020-00629-8

BACKGROUND: This study reports the analytical sensitivity and specificity of a Loop-mediated isothermal amplification (LAMP) and compares its amplification performance with conventional PCR, nested PCR (nPCR) and real-time PCR (qPCR). All the assays demonstrated in this study were developed based on Serine-rich Entamoeba histolytica protein (SREHP) gene as study model.
RESULTS: A set of SREHP gene specific LAMP primers were designed for the specific detection of Entamoeba histolytica. This set of primers recorded 100% specificity when it was evaluated against 3 medically important Entamoeba species and 75 other pathogenic microorganisms. These primers were later modified for conventional PCR, nPCR and qPCR applications. Besides, 3 different post-LAMP analyses including agarose gel electrophoresis, nucleic acid lateral flow immunoassay and calcein-manganese dye techniques were used to compare their limit of detection (LoD). One E. histolytica trophozoite was recorded as the LoD for all the 3 post-LAMP analysis methods when tested with E. histolytica DNA extracted from spiked stool samples. In contrast, none of the PCR method outperformed LAMP as both qPCR and nPCR recorded LoD of 100 trophozoites while the LoD of conventional PCR was 1000 trophozoites.
CONCLUSIONS: The analytical sensitivity comparison among the conventional PCR, nPCR, qPCR and LAMP reveals that the LAMP outperformed the others in terms of LoD and amplification time. Hence, LAMP is a relevant alternative DNA-based amplification platform for sensitive and specific detection of pathogens.

Matched MeSH terms: Nucleic Acids
Fulltext Review article : hepatitis A virus – a general overview

Yong HT, Son R

International Food Research Journal, 2009;16(4):455-467.
MyJurnal

Hepatitis A virus infection occurs globally and is causing a public health concern, primarily in developing countries due to its persistent circulation in the environment. The improved sanitary condition and increase in awareness of personal hygiene have led to the marked reduction of HAV prevalence in industrialized countries during childhood and to a shift of the infection towards adulthood. HAV is an environmentally stable, positive single stranded RNA virus that is primarily transmitted by the fecal-oral route, person to person contact or ingestion of contaminated food and drink. One of the main causes leading to HAV infection is epidemiologically linked to the consumption of raw or undercooked shellfish particularly oysters and clams. Due to their filter-feeding style, these shellfishes readily concentrate viruses from the surrounding water containing municipal sewage, and as a consequence pose a health threat to consumers. Therefore, development of detection techniques possessing the requisite sensitivity and specificity for the practical routine monitoring purposes is of great importance necessary for the protection of shellfish-consuming public. Nucleic acid based method such as reverse transcription PCR has emerged as the popular method of choice in view of its rapidity, accuracy and
sensitivity in contrary of the time-consuming conventional cell culture and hybridization techniques. However, detection of hepatitis A virus is firstly hampered by the non-cytophatic effect of wild type HAV strain, secondly, the low concentration of viral genome present in the environmental sample which requires effective isolation and concentration of virions and lastly the labor-extensive purification and thorough removal of the abundance of the PCR inhibitors which will unfavorably reduce the efficiency of PCR detection.

Matched MeSH terms: Nucleic Acids
Fulltext SINGLE NUCLEOTIDE POLYMORPHISM OF ECCB5 GENE OF MYCOBACTERIUM TUBERCULOSIS COMPLEX ISOLATES FROM SUSPECTED PULMONARY TB PATIENTS IN SURABAYA INDONESIA

Kurniawati S, Soedarsono S, Aulanni'am A, Mertaniasih NM

Afr J Infect Dis, 2018;12(2):37-42.
PMID: 30109284 DOI: 10.21010/ajid.v12i2.6

Background: Mycobacterium tuberculosis Complex (MTBC) is a group of Mycobacterium that causes tuberculosis (TB). TB is an infectious disease that remains a global health problem. Indonesia is one of the five countries in the world where TB is the most prevalent and became the country with tle second largest rate of TB in 2014 and 2015. MTBC has high pathogenicity that can cause infections in animals and humans. The most common route of transmission is via airborne droplet nuclei and contact with animals or humans infected with TB. MTBC has many virulence factors. One of these factors is EccB5 that is encoded by eccB5 gene. EccB5 is a transmembrane protein-conserved membrane protein and could play a role in inducing damage in host cells, macrophage infection, and may correlate with active disease. The characterization of eccB5 gene needs to be studied to determine the nucleotide sequences, which may be associated with active disease. The aim of this research was to analyze the nuclotide sequences of eccB5 gene of MTBC from suspected pulmonary tuberculosis patients, SNPs of eccB5 gene and possible correlation with the disease, especially in Indonesia.
Materials and Methods: Samples were collected from the Tuberculosis Laboratory, Clinical Microbiology of Dr. Soetomo Hospital Surabaya Indonesia. DNA extraction used boiling extraction method and continued nucleic acid amplification using PCR techniques. Primer pairs used eccB5 SK.. The positivity of DNA specific revealed amplicon in 1592 bp. PCR product was sequenced by 1st Base (First BASE Laboratories Sdn Bhd, Selangor, Malaysia). The sequence analysis used Genetyx-Win version 10.0 (Genetyx Corporation, Tokyo, Japan).
Results: Total isolates of Mycobacterium spp. were 28 and those that showed positive MTBC were 24 isolates and 4 nontuberculosis mycobacteria (NTM) using immunochromatographic test (ICT). The amount of homology from MTBC using blast NCBI was 99%-100%. Two SNPs were found in position c.1277 which revealed replacement of amino acid in 426 of codon position.
Conclusion: The sequence of eccB5 gene of MTBC showed high significant homology, while proposed non-synoymous single nucleotide polymorphisms (nsSNP) may associated with clinical outcomes.

Matched MeSH terms: Nucleic Acids
Nipah virus disease: A rare and intractable disease

Banerjee S, Gupta N, Kodan P, Mittal A, Ray Y, Nischal N, et al.

Intractable Rare Dis Res, 2019 Feb;8(1):1-8.
PMID: 30881850 DOI: 10.5582/irdr.2018.01130

Nipah virus, an enveloped ribonucleic acid virus, has been a major cause of encephalitis out-breaks with high mortality, primarily in the Indo-Bangladesh regions. Except for the first outbreak in Malaysia-Singapore, which was related to contact with pigs and the outbreak in Philippines associated with horse slaughter, most other outbreaks have affected the Indo- Bangladesh regions. The Indo-Bangladesh outbreaks were associated with consumption of raw date palm sap contaminated by fruit bats and had a very high secondary attack rate. The patient usually presents with fever, encephalitis and/or respiratory involvement with or without thrombocytopenia, leukopenia and transaminitis. Diagnosis can be confirmed by isolation and nucleic acid amplification in the acute phase or antibody detection during the convalescent phase. Treatment is mostly limited to supportive care and syndromic management of acute encephalitis syndrome. Ribavirin, m102.4 monoclonal antibody and favipiravir are the only anti-virals with some activity against Nipah virus. Standard precautions, hand hygiene and personal protective equipments are the cornerstone of comprehensive infection prevention and control strategy. With the recent outbreaks affecting newer geographical areas, there is a need for physicians to be aware of this disease and keep abreast of its current detection and management strategies.

Matched MeSH terms: Nucleic Acids
An iron oxide nanoworm hybrid on an interdigitated microelectrode silica surface to detect abdominal aortic aneurysms

Yan G, Li Q, Hong X, Gopinath SCB, Anbu P, Li C, et al.

Mikrochim Acta, 2021 05 11;188(6):185.
PMID: 33977395 DOI: 10.1007/s00604-021-04836-8

An abdominal aortic aneurysm (AAA) is abnormal swelling in the abdominal aorta and a prevalent life-threatening disease. This research introduces a new interdigitated microelectrode (IDME)-sensing surface modified by iron oxide nanoworms (IONWs) for detecting the AAA biomarker insulin-like growth factor-1 (IGF1). A sandwich pattern was formulated with the IGF1 aptamer and IGFBP1 (IGF binding protein-1) on the IONW-constructed IDME hybrid to identify IGF1. The surface morphology of the IONWs revealed a uniform distribution of worm-like structures (80-100 nm) as confirmed by FESEM and FETEM analyses. Further, the presence of the major elements, Fe and O, was confirmed by EDX and XPS studies. The crystal planes that appeared in the IONW reflect cubic magnetite. IONW-modified IDME attained a limit of detection for IGF1 of 1 fM (3σ) with an aptamer-IGF1-IGFBP1 sandwich. This sandwich with IGFBP1 enhanced the current level at all concentrations of IGF1 and displayed linearity in the range 1 fM to 100 pM with a determination coefficient of R2 = 0.9373 [y = 3.38221x - 4.79]. Control experiments with complementary aptamer sequences, IGF2 and IGFBP3 did not show notable signal changes, indicating the specific detection of IGF1. This IONW constructed electrode helps to achieve the detection of low amounts of IGF1 and diagnose AAA at the stage prior to rupture.

Matched MeSH terms: Immobilized Nucleic Acids/chemistry
Electrochemical determination of zearalenone using a label-free competitive aptasensor

Azri FA, Eissa S, Zourob M, Chinnappan R, Sukor R, Yusof NA, et al.

Mikrochim Acta, 2020 04 12;187(5):266.
PMID: 32279134 DOI: 10.1007/s00604-020-4218-7

An electrochemical aptasensor is described for determination of the phytohormone of zearalenone (ZEA). The gold electrode was modified with ZEA via covalent attachment using cysteamine-hydrochloride and 1,4-phenylene diisocyanate linker. A truncated ZEA aptamer with a dissociation constant of 13.4 ± 2.1 nM was used in an aptasensor. The electrochemical property was investigated using square wave voltammetry for monitoring the change in the electron transfer using the ferro/ferricyanide system as redox probe. Under optimal experimental conditions, the response was best measured at a potential of 0.20 V (vs. Ag/AgCl). The signals depended on the competitive mechanism between the immobilised ZEA and free ZEA for the aptamer binding site. The aptasensor works in the range 0.01 to 1000 ng·mL-1 ZEA concentration, with a detection limit of 0.017 ng·mL-1. High degree of cross-reactivity with the other analogues of ZEA was observed, whereas none towards other mycotoxins. The aptasensor was further applied for the determination of ZEA in the extract of maize grain and showed good recovery percentages between 87 and 110%. Graphical abstract Schematic representation of the electrochemical determination of zearalenone based on indirect competitive assay. Step a Immobilisation of ZEA on the surface of gold electrode via covalent attachment, b competition for the ZEA aptamer binding site between immobilised and free ZEA, and c current signal of the binding event based on SWV technique.

Matched MeSH terms: Immobilized Nucleic Acids/chemistry
Cellulose supported magnetic nanohybrids: Synthesis, physicomagnetic properties and biomedical applications-A review

Haniffa MACM, Munawar K, Chee CY, Pramanik S, Halilu A, Illias HA, et al.

Carbohydr Polym, 2021 Sep 01;267:118136.
PMID: 34119125 DOI: 10.1016/j.carbpol.2021.118136

Cellulose and its forms are widely used in biomedical applications due to their biocompatibility, biodegradability and lack of cytotoxicity. It provides ample opportunities for the functionalization of supported magnetic nanohybrids (CSMNs). Because of the abundance of surface hydroxyl groups, they are surface tunable in either homogeneous or heterogeneous solvents and thus act as a substrate or template for the CSMNs' development. The present review emphasizes on the synthesis of various CSMNs, their physicomagnetic properties, and potential applications such as stimuli-responsive drug delivery systems, MRI, enzyme encapsulation, nucleic acid extraction, wound healing and tissue engineering. The impact of CSMNs on cytotoxicity, magnetic hyperthermia, and folate-conjugates is highlighted in particular, based on their structures, cell viability, and stability. Finally, the review also discussed the challenges and prospects of CSMNs' development. This review is expected to provide CSMNs' development roadmap in the context of 21st-century demands for biomedical therapeutics.

Matched MeSH terms: Nucleic Acids/isolation & purification
Improved sensitivity of lateral flow assay using paper-based sample concentration technique

Tang R, Yang H, Choi JR, Gong Y, Hu J, Feng S, et al.

Talanta, 2016 May 15;152:269-76.
PMID: 26992520 DOI: 10.1016/j.talanta.2016.02.017

Lateral flow assays (LFAs) hold great promise for point-of-care testing, especially in resource-poor settings. However, the poor sensitivity of LFAs limits their widespread applications. To address this, we developed a novel device by integrating dialysis-based concentration method into LFAs. The device successfully achieved 10-fold signal enhancement in Human Immunodeficiency Virus (HIV) nucleic acid detection with a detection limit of 0.1nM and 4-fold signal enhancement in myoglobin (MYO) detection with a detection limit of 1.56ng/mL in less than 25min. This simple, low-cost and portable integrated device holds great potential for highly sensitive detection of various target analytes for medical diagnostics, food safety analysis and environmental monitoring.

Matched MeSH terms: Nucleic Acids

Filters

Please provide feedback to Administrator (afdal@afpm.org.my)

External Links