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The effect of extrinsic incubation temperature on development of dengue serotype 2 and 4 viruses in Aedes aegypti (L.)

Rohani A, Wong YC, Zamre I, Lee HL, Zurainee MN

Southeast Asian J. Trop. Med. Public Health, 2009 Sep;40(5):942-50.
PMID: 19842378

Dengue 2 and 4 viruses obtained from dengue-infected patients were maintained in a C6/36 Aedes albopictus Skuse cell line and used to infect adult female Aedes aegypti mosquitoes. Each serotype was mixed separately with fresh human erythrocytes and fed to adult female mosquitoes using an artificial membrane feeding technique. Fully engorged mosquitoes were selected and retained at 26 degrees C, 28 degrees C and 30 degrees C to observe dengue virus development in Aedes vectors. Virus detection was carried out by reverse-transcriptase polymerase chain reaction (RT-PCR). The virus was first detected on Day 9 at 26 degrees C and 28 degrees C and on Day 5 at 30 degrees C for both dengue 2 and 4. The study shows the incubation period of the viruses decreased when the extrinsic incubation temperature increases.

Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction
Fulltext Risk factors and spatial distribution of extended spectrum β-lactamase-producing- Escherichia coli at retail poultry meat markets in Malaysia: a cross-sectional study

Aliyu AB, Saleha AA, Jalila A, Zunita Z

BMC Public Health, 2016 08 02;16:699.
PMID: 27484086 DOI: 10.1186/s12889-016-3377-2

BACKGROUND: The significant role of retail poultry meat as an important exposure pathway for the acquisition and transmission of extended spectrum β-lactamase-producing Escherichia coli (ESBL-EC) into the human population warrants understanding concerning those operational practices associated with dissemination of ESBL-EC in poultry meat retailing. Hence, the objective of this study was to determine the prevalence, spatial distribution and potential risk factors associated with the dissemination of ESBL-EC in poultry meat retail at wet-markets in Selangor, Malaysia.
METHODS: Poultry meat (breast, wing, thigh, and keel) as well as the contact surfaces of weighing scales and cutting boards were sampled to detect ESBL-EC by using culture and disk combination methods and polymerase chain reaction assays. Besides, questionnaire was used to obtain data and information pertaining to those operational practices that may possibly explain the occurrence of ESBL-EC. The data were analysed using logistic regression analysis at 95 % CI.
RESULTS: The overall prevalence of ESBL-EC was 48.8 % (95 % CI, 42 - 55 %). Among the risk factors that were explored, type of countertop, sanitation of the stall environment, source of cleaning water, and type of cutting board were found to be significantly associated with the presence of ESBL-EC.
CONCLUSIONS: Thus, in order to prevent or reduce the presence of ESBL-EC and other contaminants at the retail-outlet, there is a need to design a process control system based on the current prevailing practices in order to reduce cross contamination, as well as to improve food safety and consumer health.

Matched MeSH terms: Polymerase Chain Reaction
Characterization of secreted recombinant Toxoplasma gondii surface antigen 2 (SAG2) heterologously expressed by the yeast Pichia pastoris

Fong MY, Lau YL, Zulqarnain M

Biotechnol Lett, 2008 Apr;30(4):611-8.
PMID: 18043869

The surface antigen 2 (SAG2) gene of the protozoan parasite, Toxoplasma gondii, was cloned and extracellularly expressed in the yeast Pichia pastoris. The effectiveness of the secreted recombinant SAG2 (rSAG2-S) as a serodiagnosis reagent was assessed by western blots and ELISA. In the western blot assay, rSAG2-S reacted with all Toxoplasma-antibody positive human serum samples but not with Toxoplasma-negative samples. In the ELISA, rSAG2-S yielded sensitivity rates ranging from 80% (IgG negative, IgM positive) to 100% (IgG positive, IgM negative). In vivo experiments showed that serum from mice immunized with rSAG2-S reacted specifically with the native SAG2 of T. gondii. These mice were protected when challenged with live cells of T. gondii.

Matched MeSH terms: Polymerase Chain Reaction
Fulltext A review on the global widespread of TTV infection among humans population

Nur Syazwani Jarkasi, Zamberi Sekawi, Cheah, Yoke Kqueen, Zulkefley Othman

The Pertanika Journal of Scholarly Research Reviews, 2018;4(1):10-24.
MyJurnal

Torque Teno Virus (TTV) is a human-infected virus that is present ubiquitously in nature. Globally, it infects up to 95% of the healthy individuals without any clinical manifestations. The widely used laboratory diagnosis of TTV infection is Polymerase chain reaction (PCR). Nevertheless, several other methods have been developed. The rapid growth of TTV variants over time has posed a challenge in estimating the global TTV infection as none of the PCR protocol has the ability to detect the entire spectrum of TTV variants. Multiple TTV epidemiological studies have been conducted among Asian population, whereas other continents showed a limited number of studies. The horizontal and vertical transmission of TTV among humans population, as well as interspecies transmission are potentially related to the global widespread of TTV infection.

Matched MeSH terms: Polymerase Chain Reaction
Fulltext Light microscopy and molecular identification of Sarcocystis spp. in meat producing animals in Selangor, Malaysia

Latif B, Kannan Kutty M, Muslim A, Hussaini J, Omar E, Heo CC, et al.

Trop Biomed, 2015 Sep;32(3):444-52.
PMID: 26695204 MyJurnal

One thousand and forty-five tissue samples of skeletal muscles, tongue, heart, diaphragm and esophagus were collected from 209 animals (43 sheep, 89 goats and 77 cattle) from an abattoir in Selangor between February and October, 2013. Each sample was divided into three pieces with each piece measuring 2-3 mm3. Each piece was then squeezed between two glass slides and examined microscopically at x 10 magnification for the presence of sarcocystosis. Three positive samples from each animal species were then fixed in 10% formalin for histological processing. Seven positive samples collected from each animal species were preserved at -80°C or 90% ethanol for gene expression studies. Microsarcocysts were detected in 114 (54.5%) animals by light microscopy (LM). The infection rates in sheep, goat and cattle were 86, 61.8 and 28.6% respectively. The highest rate of infection was in the skeletal muscles of sheep (64.9%) and goats (63.6%) and in the heart of cattle (63.6%). The cysts were spindle to oval in shape and two stages were recognized, the peripheral metrocytes and centrally located banana-shaped bradyzoites. 18S rRNA gene expression studies confirmed the isolates from the sheep as S. ovicanis, goats as S. capracanis and cattle as S. bovicanis. This, to the best of our knowledge, is the first molecular identification of an isolate of S. ovicanis and S. capracanis in Malaysia. Further studies with electron microscopy (EM) are required in the future to compare the features of different types of Sarcocysts spp.

Matched MeSH terms: Polymerase Chain Reaction
Fulltext Sybr Green Real-Time PCR is a useful method in screening for high risk human papilloma virus types 16 and 18 in neoplastic cervical cancer

Nor Rizan, K., Abdul Manaf ,A., Sabariah, A.R., Siti Aishah, M.A., Noorjahan Banu, M.A., Zubaidah, Z.

Medicine & Health, 2011;6(1):59-67.
MyJurnal

Human papillomavirus (HPV) plays an important role in the pathogenesis of cervical cancer. HPV has been found in 99.7% of cervical cancers worldwide. In Malaysia, it is the second most common cancer among women in all major ethnic groups. The main purpose of this study was to establish the method of SyBrGreen Real-Time PCR and apply it for identification of multiple infections of the two high risk HPV subtypes. In this study, 57 positive samples for HPV 16 and HPV 18 were used to establish a simple
and sensitive method to detect and identify HPV infection in the cervical neoplasia at different stages of the disease by using real-time ABICycler SyBrGreen 1 technology. The results showed 67 HPV genomes in 57 samples. HPV 16 genome was detected in 55/67 (82%) cases while HPV 18 was detected in 8/67 (12%) cases with 4 cases showing multiple infections of HPV 16 and HPV 18. HPV 16 was the most prevalent followed by HPV 18. Using SyBr Green Real-Time PCR techniques, the results
showed that DNA melting curve for HPV 16 had a peak around 80.2ºC and Ct value of 20 cycles whereas the DNA melting curve for HPV 18 around 79.2ºC and Ct value of 22 cycles. In conclusion, a SyBr Green Real-Time PCR method has the potential for clinical usage in detection and identification of HPV infection in cervical neoplasia at different stages of the disease.

Matched MeSH terms: Real-Time Polymerase Chain Reaction
Real time quantitative PCR by the LightCycler system

Chin, Yuet Meng, Arison Mohamad, Zubaidah Zakaria

Buletin Persatuan Genetik Malaysia, 2002;8(1):0-0.
MyJurnal

For many years counting cells and identifying them under the microscope has been the conventional method to determine the number of abnormal and normal cells in cancers. During the last decade, studies have shown that the detection and quantification of residual tumor cells is important in predicting the clinical outcome of several types of hematological malignancies. Detection of
minimal residual disease (MRD) is now becoming routinely implemented in treatment protocols and is increasingly used for guiding therapy and for evaluation of new treatment modalities (Raanani & Hashomer, 2004). A wide variety of techniques have been developed to detect residual malignant cells beyond the sensitivity of conventional approaches by cell morphology. One of these technology is by real time quantitative (RQ) polymerase chain reaction (PCR) using the Taqman and LightCycler systems.

Matched MeSH terms: Real-Time Polymerase Chain Reaction
Fulltext Distribution of alpha thalassaemia in 16 year old Malaysian Students in Penang, Melaka and Sabah

Rahimah AN, Nisha S, Safiah B, Roshida H, Punithawathy Y, Nurul H, et al.

Med J Malaysia, 2012 Dec;67(6):565-70.
PMID: 23770946 MyJurnal

OBJECTIVES: Alpha thalassaemia is wide spread in Malaysia and is a public health problem. This study aimed to describe the carrier frequencies of α‒thalassaemia and its distribution among major ethnic groups in three states of Malaysia.

METHODS: Educational forums were organised and study was explained to students from three schools. Students were invited to take part in the screening with parent consent. A total of 8420 adolescent students aged 16 years volunteered to participate in the study. Peripheral blood samples were analysed for complete blood counts, haemoglobin quantification and typing, and serum ferritin levels. Genomic DNA was used for screening alpha thalassaemia alleles by PCR based molecular methods.

RESULTS: We identified seven α‒globin gene defects in 341 (4.08%) students: amongst them α(+)‒ and α(0)‒thalassaemias were detected in 232 (2.77%) and 107 (1.28%) students respectively. Genotype ‒α(3.7)/αα was the most prevalent among sub-populations of Malay, indigenous communities of Sahab and Indian, while ‒‒(SEA)/αα deletion is more prevalent in Malaysian Chinese. It is estimated that 63 pregnancies annually are at risk of Hb Bart's hydrops fetalis.

CONCLUSIONS: We have demonstrated the prevalence and mutation patterns of α‒thalassaemia in the 16 year olds in three states of Malaysia. High α(0)‒thalassaemia deletions amongst the study subjects place these carriers at an increased risk of conceiving fetuses with HbH disease and Hb Bart's hydrops fetalis should they choose another heterozygous partner. It is therefore highly recommended to institute community screening programmes and provide prospective carriers with genetic counselling to help them make informed choices.

Matched MeSH terms: Polymerase Chain Reaction
Fulltext Diagnostic techniques for COVID-19 and new developments

Sheikhzadeh E, Eissa S, Ismail A, Zourob M

Talanta, 2020 Dec 01;220:121392.
PMID: 32928412 DOI: 10.1016/j.talanta.2020.121392

COVID-19 pandemic is a serious global health issue today due to the rapid human to human transmission of SARS-CoV-2, a new type of coronavirus that causes fatal pneumonia. SARS -CoV-2 has a faster rate of transmission than other coronaviruses such as SARS and MERS and until now there are no approved specific drugs or vaccines for treatment. Thus, early diagnosis is crucial to prevent the extensive spread of the disease. The reverse transcription-polymerase chain reaction (RT-PCR) is the most routinely used method until now to detect SARS-CoV-2 infections. However, several other faster and accurate assays are being developed for the diagnosis of COVID-19 aiming to control the spread of infection through the identification of patients and immediate isolation. In this review, we will discuss the various detection methods of the SARS-CoV-2 virus including the recent developments in immunological assays, amplification techniques as well as biosensors.

Matched MeSH terms: Polymerase Chain Reaction
Fulltext Posterior capsule rupture-causes, associations and outcomes: eight year analysis in a Malaysian General Hospital

Thevi T, Zin MM

International Eye Science, 2016;16:600-606.

AIM: To see the causative factors, associations and outcomes of posterior capsule rupture (PCR). METHODS: A retrospective cohort study was done of all patients with PCR from 2007 to 2014 in Melaka Hospital. Associations between ocular comorbidities, lens related complications, surgeon grade, type of cataract surgery, whether done alone or in combination, with the occurrences of PCR were studied. The final visual outcome of cases with PCR was studied. RESULTS: PCR was the commonest intraoperative complication (n=623, 4.8%) among 12 846 patients. Ocular comorbidities and status of the lens did not cause PCR. Experience and seniority of surgeons were significantly associated with PCR. Medical officers had more PCRs than gazetting specialists (P=0.0000), who inturn had more PCRs than specialists (P=0.000). Each type of cataract surgery done was also significantly associated with PCR-phacoemulsification (phaco), phaco convert to extracapsular cataract extraction (ECCE) and intracapsular cataract extraction (ICCE) (P=0.000), lens aspiration (P=0.020), and ECCE (P=0.003). Specialists got good outcomes compared to trainees without PCR occurring (P=0.001) and also with PCR occurring (P=0.013). However, no difference was observed in the occurrence of PCR in complicated cases between specialists and trainees. Vision was compromised mainly by astigmatism following PCR. Impaired vision (P=0.000) and poor vision (P=0.000) were more than good vision. Poor vision was more in PCR compared to other complications.(P=0.000). CONCLUSION: PCR occurs in all types of cataract surgeries and is the main intraoperative complication causing poor vision. Good outcomes were significantly more when specialists got PCR compared to trainees. Junior surgeons should practice in wet labs and be given more cases. Copyright 2016 by the IJO Press.

Matched MeSH terms: Polymerase Chain Reaction
The USM human genome centre experience on molecular diagnostic testing of spinal muscular atrophy

Fatemeh, H., Watihayati, M.S., Marini, M., NurShafawati, A.R., Atif, A.B., Zabidi-Hussin, Z.A.M.H., et al.

Malaysian Journal of Paediatrics and Child Health, 2007;15(1):24-0.
MyJurnal

Spinal Muscular Atrophy (SMA) is a heredity neuromuscular disorder and is one of the most common genetic causes of childhood fatality. SMA is classified into three groups based on age of onset and achieved motor milestone. Survival Motor Neuron (SMN) gene has been identified as the responsible gene for SMA. From August 2003 until Feb 2007 we have received 93 samples for SMN1 gene deletion analysis from various hospitals in Malaysia. All the patients except for 3 patients were Malaysian (71 Malays, 5 Indians, 9 Chinese and 5 patients are mixed ethnicity). DNA were extracted from blood samples using DNA extraction kit and subjected to SMN/ gene deletion analysis by PCR-RE. Forty nine out of 93 samples (20 type I, 21 type II, and 8 type III) were found to have homozygous deletion of at least exon 7 of the SMN1 gene. Twelve patients (7 type I, 4 type II, 1 type III) showed the presence of the SMN1 gene and the rest were excluded as they did not fulfill the criteria of International SMA Consortium. Deletion analysis of exon 7 of the SMN gene can be an alternative to the existing diagnostic modalities of SMA.

Matched MeSH terms: Polymerase Chain Reaction
A new method for FMR1 gene methylation screening by multiplex methylation-specific real-time polymerase chain reaction

Elias MH, Ankathil R, Salmi AR, Sudhikaran W, Limprasert P, Zilfalil BA

Genet Test Mol Biomarkers, 2011 Jun;15(6):387-93.
PMID: 21329465 DOI: 10.1089/gtmb.2010.0191

Fragile X Syndrome (FXS) is the most common form of inherited mental retardation in men. It is caused by abnormalities in the FMR1 gene that are associated with CGG repeat expansion and the hypermethylation status of its promoter. Methylated alleles lead to transcriptional inhibition and consequent loss of Fragile X Mental Retardation Protein. Chemical modification of cytosine to uracil by bisulfite treatment has proved to be an attractive method for DNA methylation studies that precludes labor-intensive Southern blot analysis, which is the gold standard test for FXS. In this report, bisulfite-treated DNA samples were amplified using real-time multiplex methylation-specific polymerase chain reaction followed by melting curve analysis. Our results show that all control samples with known CGG repeat numbers and methylation statuses were correctly diagnosed. The samples from 43 male patients were also successfully diagnosed, which were in complete agreement with the results of Southern blotting. By such means, 39 patients were found to have an unmethylated allele; 3, a fully mutated allele; and 1, both methylated and unmethylated alleles, thus implying a diagnosis of mosaicism. In conclusion, we find our method to be convenient for screening a large number of male patients with FXS, because it is rapid and easy to perform, especially when there is a low quantity of DNA that must be sensitively and accurately assayed.

Matched MeSH terms: Polymerase Chain Reaction/methods*
Screening for gap junction protein beta-2 gene mutations in Malays with autosomal recessive, non-syndromic hearing loss, using denaturing high performance liquid chromatography

Aishah ZS, Khairi MD, Normastura AR, Zafarina Z, Zilfalil BA

J Laryngol Otol, 2008 Dec;122(12):1284-8.
PMID: 18353197 DOI: 10.1017/S0022215108002041

To determine the frequency and type of gap junction protein beta-2 gene mutations in Malay patients with autosomal recessive, non-syndromic hearing loss.

Matched MeSH terms: Polymerase Chain Reaction
Combination of SMN2 copy number and NAIP deletion predicts disease severity in spinal muscular atrophy

Watihayati MS, Fatemeh H, Marini M, Atif AB, Zahiruddin WM, Sasongko TH, et al.

Brain Dev, 2009 Jan;31(1):42-5.
PMID: 18842367 DOI: 10.1016/j.braindev.2008.08.012

Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder caused by mutations in the SMN1 gene. The SMN2 gene is highly homologous to SMN1 and has been reported to be correlated with severity of the disease. The clinical presentation of SMA varies from severe to mild, with three clinical subtypes (type I, type II, and type III) that are assigned according to age of onset and severity of the disease. Here, we aim to investigate the potential association between the number of copies of SMN2 and the deletion in the NAIP gene with the clinical severity of SMA in patients of Malaysian origin. Forty-two SMA patients (14 of type I, 20 type II, and 8 type III) carrying deletions of the SMN1 gene were enrolled in this study. SMN2 copy number was determined by fluorescence-based quantitative polymerase chain reaction assay. Twenty-nine percent of type I patients carried one copy of SMN2, while the remaining 71% carried two copies. Among the type II and type III SMA patients, 29% of cases carried two copies of the gene, while 71% carried three or four copies of SMN2. Deletion analysis of NAIP showed that 50% of type I SMA patients had a homozygous deletion of exon 5 of this gene and that only 10% of type II SMA cases carried a homozygous deletion, while all type III patients carried intact copies of the NAIP gene. We conclude that there exists a close relationship between SMN2 copy number and SMA disease severity, suggesting that the determination of SMN2 copy number may be a good predictor of SMA disease type. Furthermore, NAIP gene deletion was found to be associated with SMA severity. In conclusion, combining the analysis of deletion of NAIP with the assessment of SMN2 copy number increases the value of this tool in predicting the severity of SMA.

Matched MeSH terms: Polymerase Chain Reaction/methods
Fulltext Detection of beta-globin gene mutations among Kelantan Malay thalassaemia patients by polymerase chain reaction restriction fragment length polymorphism

Rozitah R, Nizam MZ, Nur Shafawati AR, Nor Atifah MA, Dewi M, Kannan TP, et al.

Singapore Med J, 2008 Dec;49(12):1046-9.
PMID: 19122960

Beta-thalassaemia major is an autosomal recessive disorder that results in severe microcytic, hypochromic, haemolytic anaemia among affected patients. Beta-thalassaemia has emerged as one of the most common public health problems in Malaysia, particularly among Malaysian Chinese and Malays. This study aimed to observe the spectrum of mutations found in Kelantan Malay beta-thalassaemia major patients who attended the Paediatrics Daycare Unit, Hospital Universiti Sains Malaysia, Kelantan, Malaysia, the data of which was being used in establishing the prenatal diagnosis in this Human Genome Centre.

Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction
Allele-specific PCR for a cost-effective & time-efficient diagnostic screening of spinal muscular atrophy

Marini M, Sasongko TH, Watihayati MS, Atif AB, Hayati F, Gunadi, et al.

Indian J Med Res, 2012;135:31-5.
PMID: 22382180

Genetic diagnosis of spinal muscular atrophy (SMA) is complicated by the presence of SMN2 gene as majority of SMA patients show absence or deletion of SMN1 gene. PCR may amplify both the genes non selectively in presence of high amount of DNA. We evaluated whether allele-specific PCR for diagnostic screening of SMA is reliable in the presence of high amount of genomic DNA, which is commonly used when performing diagnostic screening using restriction enzymes.

Matched MeSH terms: Polymerase Chain Reaction/methods*
PCR tools for the verification of the specific identity of ascaridoid nematodes from dogs and cats

Li MW, Lin RQ, Chen HH, Sani RA, Song HQ, Zhu XQ

Mol Cell Probes, 2007 Oct-Dec;21(5-6):349-54.
PMID: 17532185

Based on the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of Toxocara canis, Toxocara cati, Toxocara malaysiensis and Toxascaris leonina, specific forward primers were designed in the ITS-1 or ITS-2 for each of the four ascaridoid species of dogs and cats. These primers were used individually together with a conserved primer in the large subunit of rDNA to amplify partial ITS-1 and/or ITS-2 of rDNA from 107 DNA samples from ascaridoids from dogs and cats in China, Australia, Malaysia, England and the Netherlands. This approach allowed their specific identification, with no amplicons being amplified from heterogeneous DNA samples, and sequencing confirmed the identity of the sequences amplified. The minimum amounts of DNA detectable using the PCR assays were 0.13-0.54ng. These PCR assays should provide useful tools for the diagnosis and molecular epidemiological investigations of toxocariasis in humans and animals.

Matched MeSH terms: Polymerase Chain Reaction/methods*
Molecular surveillance of coxsackievirus A16 reveals the emergence of a new clade in mainland China

Chen L, Yao XJ, Xu SJ, Yang H, Wu CL, Lu J, et al.

Arch Virol, 2018 Nov 29.
PMID: 30498962 DOI: 10.1007/s00705-018-4112-3

Coxsackievirus A16 (CV-A16) of the genotypes B1a and B1b have co-circulated in mainland China in the past decades. From 2013 to 2017, a total of 3,008 specimens from 3,008 patients with mild hand, foot, and mouth disease were collected in the present study. Viral RNA was tested for CV-A16 by a real-time RT-PCR method, and complete VP1 sequences and full-length genome sequences of CV-A16 strains from this study were determined by RT-PCR and sequencing. Sequences were analyzed using a series of bioinformatics programs. The detection rate for CV-A16 was 4.1%, 25.9%, 10.6%, 28.1% and 12.9% in 2013, 2014, 2015, 2016 and 2017, respectively. Overall, the detection rate for CV-A16 was 16.5% (497/3008) in this 5-year period in Shenzhen, China. One hundred forty-two (142/155, 91.6%) of the 155 genotype B1 strains in the study belonged to subgenotype B1b, and 13 (13/155, 8.4%) strains belonged to subgenotype B1a. Two strains (CVA16/Shenzhen174/CHN/2017 and CVA16/Shenzhen189/CHN/2017) could not be assigned to a known genotype. Phylogenetic analysis of these two strains and other Chinese CV-A16 strains indicated that these two CV-A16 strains clustered independently in a novel clade whose members differed by 8.4%-11.8%, 8.4%-12.1%, and 14.6%-14.8% in their nucleotide sequences from those of Chinese B1a, B1b, and genotype D strains, respectively. Phylogenetic analysis of global CV-A16 strains further indicated that the two novel CV-A16 strains from this study grouped in a previously uncharacterized clade, which was designated as the subgenogroup B3 in present study. Meanwhile, phylogenetic reconstruction revealed two other new genotypes, B1d and B4, which included a Malaysian strain and two American strains, respectively. The complete genome sequences of the two novel CV-A16 strains showed the highest nucleotide sequence identity of 92.3% to the Malaysian strain PM-15765-00 from 2000. Comparative analysis of amino acid sequences of the two novel CV-A16 strains and their relatives suggested that variations in the nonstructural proteins may play an important role in the evolution of modern CV-A16.

Matched MeSH terms: Reverse Transcriptase Polymerase Chain Reaction; Real-Time Polymerase Chain Reaction
Decellularized Matrix Derived from Neural Differentiation of Embryonic Stem Cells Enhances the Neurogenic Potential of Dental Follicle Stem Cells

Heng BC, Gong T, Wang S, Lim LW, Wu W, Zhang C

J Endod, 2017 Mar;43(3):409-416.
PMID: 28231979 DOI: 10.1016/j.joen.2016.10.033

INTRODUCTION: Dental follicle stem cells (DFSCs) possess neurogenic potential because they originate from the embryonic neural crest. This study investigated whether neural differentiation of DFSCs can be enhanced by culture on decellularized matrix substrata (NSC-DECM) derived from neurogenesis of human embryonic stem cells (hESCs).
METHODS: The hESCs were differentiated into neural stem cells (NSCs), and NSC-DECM was extracted from confluent monolayers of NSCs through treatment with deionized water. DFSCs seeded on NSC-DECM, Geltrex, and tissue culture polystyrene (TCPS) were subjected to neural induction during a period of 21 days. Expression of early/intermediate (Musashi1, PAX6, NSE, and βIII-tubulin) and mature/late (NGN2, NeuN, NFM, and MASH1) neural markers by DFSCs was analyzed at the 7-, 14-, and 21-day time points with quantitative real-time polymerase chain reaction. Immunocytochemistry for detection of βIII-tubulin, PAX6, and NGN2 expression by DFSCs on day 7 of neural induction was also carried out.
RESULTS: Quantitative RT-PCR showed that expression of PAX6, Musashi1, βIII-tubulin, NSE, NGN2, and NFM by DFSCs was enhanced on NSC-DECM versus either the Geltrex or TCPS groups. Immunocytochemistry showed that DFSCs in the NSC-DECM group displayed more intense staining for βIII-tubulin, PAX6, and NGN2 expression, together with more neurite outgrowths and elongated morphology, as compared with either Geltrex or TCPS.
CONCLUSIONS: DECM derived from neurogenesis of hESCs can enhance the neurogenic potential of DFSCs.

Matched MeSH terms: Real-Time Polymerase Chain Reaction
Fulltext Symptomatic chronic strongyloidiasis in children following treatment for solid organ malignancies: case reports and literature review

Norsarwany M, Abdelrahman Z, Rahmah N, Ariffin N, Norsyahida A, Madihah B, et al.

Trop Biomed, 2012 Sep;29(3):479-88.
PMID: 23018511

Strongyloidiasis is an infection caused by the intestinal nematode Strongyloides stercoralis. Infected healthy individuals are usually asymptomatic, however it is potentially fatal in immunocompromised hosts due to its capacity to cause an overwhelming hyperinfection. Strongyloidiasis could be missed during routine screening because of low and intermittent larval output in stool and variable manifestations of the symptoms. We present two cases of strongyloidiasis occurring in children with solid organ malignancies suspected to have the infection based on their clinical conditions and treatment history for cancer. Both patients were diagnosed by molecular and serological tests and were successfully treated. Thus, strongyloidiasis in patients undergoing intensive treatment for malignancies should be suspected, properly investigated and treated accordingly.

Matched MeSH terms: Real-Time Polymerase Chain Reaction

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