Human cytokeratin 1 plays a role in the interaction of Pteropine orthoreovirus with Hek293 cells but not HeLa cells.

Pteropine orthoreovirus (PRV) causes respiratory tract infections in humans. Despite its emergence as a zoonotic and respiratory virus, little is known about its cell tropism, which hampers progress in fully understanding its pathogenesis in humans. Hek293 cells are most susceptible to PRV infection, while HeLa cells are the least. Human cytokeratin 1 (CK1) was identified as the protein that interacts with PRV. The immunofluorescence assay and qPCR results revealed prior treatment with anti-CK1 may provide Hek293 cells protection against PRV. The KRT1-knockout Hek293 cells were less susceptible to PRV infection. Further study into the pathogenesis of PRV in humans is needed.

Oncolytic Reoviruses: Can These Emerging Zoonotic Reoviruses Be Tamed and Utilized?

Orthoreovirus is a nonenveloped double-stranded RNA virus under the Reoviridae family. This group of viruses, especially mammalian orthoreovirus (MRV), are reported with great therapeutic values due to their oncolytic effects. In this review, the life cycle and oncolytic effect of MRV and a few emerging reoviruses were summarized. This article also highlights the challenges and strategies of utilizing MRV and the emerging reoviruses, avian orthoreovirus (ARV) and pteropine orthoreovirus (PRV), as oncolytic viruses (OVs). Besides, the emergence of potential ARV and PRV as OVs were discussed in comparison to MRV. Finally, the risk of reovirus as zoonosis or reverse zoonosis (zooanthroponosis) were debated, and concerns were raised in this article, which warrant continue surveillance of reovirus (MRV, ARV, and PRV) in animals, humans, and the environment.

Incorporating Insulin Growth Factor-1 into Regenerative and Personalized Medicine for Cardiovascular Disease: A Systematic Review.

BACKGROUND: Cardiovascular disease (CVD) is one of the world's leading causes of increased morbidity and mortality. Current interventions for CVD, including percutaneous transluminal coronary angioplasty (PTCA) and coronary artery bypass grafting (CABG), carry certain risks and complications, which may also affect the patient's quality of life. It is important to minimize those risks and complications while speeding up the recovery. Insulin Growth Factor-1 (IGF-1) is a growth factor responsible for cellular migration, proliferation, differentiation, and angiogenesis, which supports cardiovascular regeneration. METHODS: In light of the current trend of regenerative medicine, the present review aims to pool data relating to the incorporation of IGF-1 in regenerative medicine and provide input on the current research gaps and concerns arising on translating this approach from benchwork into clinical settings. RESULTS: Using the keywords IGF-1 'OR' Insulin Growth Factor 1 'AND' Mesenchymal Stem Cells 'AND' Tissue Healing from 2009 to 2020, we identified 160 and 52 from Medline and PubMed, screening out 202 articles due to non-fulfilment of the inclusion criteria. CONCLUSION: Incorporating IGF-1 into regenerative and personalized medicine may be promising for treating CVD; however, the concerns include the role of IGF-1 in inducing cancer growth and its ability to migrate to the specific site of injury, especially for those who present with multiple pathologies should be addressed prior to its translation from bench work into clinical settings.

Pteropine Orthoreovirus, PRV7S (Sikamat Virus) Demonstrates Oncolysis in Nasopharyngeal Carcinoma Cell Lines.

BACKGROUND: Oncolytic properties had been demonstrated in Mammalian Orthoreovirus (MRV) and Avian Orthorevirus (ARV). Besides MRV and ARV, Pteropine Orthoreovirus (PRV) is also categorized under the genus Orthoreovirus. PRV7S (Sikamat virus) is an orthoreovirus isolated in Malaysia. Present study aims to investigate the oncolytic effects of PRV7S on ranges of nasopharyngeal carcinoma (NPC) cells through apoptosis in comparison to MRV3. METHODS: Non-cancerous nasopharyngeal (NCNP) and NPC cells were infected by PRV7S and MRV3. The effects of PRV7S on the proliferation inhibition and apoptotic activity of NPC cells was examined using MTT assay and flow cytometry. Additionally, western blot assay was performed to analyze the expression of RAS and apoptotic protein. Lastly, qPCR assay was performed to demonstrate that PRV7S and MRV3 replicated in infected-NPC and infected-NCNP cells. RESULTS: The proliferation of NPC cells were significantly inhibited after PRV7S infection in a time dependent manner in comparison to infected-NCPC cells. Flow cytometry analysis showed that PRV7S infection was able to induce apoptosis on NPC cells at 48 hpi. Western blot results showed that upon PRV7S infection, N/H/K RAS protein expression was reduced, whereas caspase-3 protein expression increased in NPC cells. qPCR assay showed higher viral load of PRV7S found in infected-NPC compared to infected-NCNP cells. CONCLUSIONS: PRV7S inhibits the proliferation and induces apoptosis of NPC cells similar to MRV3. Therefore, PRV7S is a potential oncolytic virus.

Knowledge, Perceptions and Behaviors Related to COVID-19 in a University Setting in Malaysia.

Background: In Malaysia the COVID-19 disease (COVID-19) has continued to escalate since its first detection in late January 2020, despite widespread implementation of control measures. This study aims to determine the knowledge, perception and behaviors with respect to COVID-19 in the midst of the third wave of the infection. Methods: A cross-sectional study was carried out among staffs and students of Universiti Tunku Abdul Rahman (UTAR). The survey consists of basic sociodemographic information, 22 items on knowledge on COVID-19, 3 items on perceived self-risk, 2 items on preparedness & perceived self-efficacy, 10 items on preventive (own) measures, 9 items assessing unwanted and desirable behaviors during the pandemic. Simple and multiple linear regression were performed to determine the factors associated with knowledge, preventive measures adopted, self-risk perception, preparedness & perceived self-efficacy, and behaviors. Results: A total of 434 responded to the survey of whom the majority (85.1%) had high scores for knowledge (mean score of 18.72 out of 22). A significant positive association was found between knowledge and older age (adjusted B coefficient (SE) = 0.046 (0.022), p = 0.039), those from medical faculty (adjusted B coefficient (SE) = 0.870 (0.420), p = 0.039) and residence in high-risk areas (adjusted B coefficient (SE) = 0.831 (0.295), p = 0.005). Predictors for higher perception of COVID-19 risk included presence of COVID-19 cases among social contacts (adjusted B coefficient (SE) = 0.751 (0.308), p = 0.015) and living with elderly (adjusted B coefficient (SE) = 1.137 (0.296), p < 0.001), while that for perception of preparedness and self-efficacy were living with children (adjusted Beta coefficient (SE) = 0.440 (0.173), p = 0.011) and absence of positive cases among social contacts (adjusted B coefficient (SE) = 0.418 (0.183), p = 0.023). Good preventive measures among the respondents were positively associated with knowledge (adjusted B coefficient (SE) = 0.116 (0.025), p < 0.001), as well as with female gender (adjusted B coefficient (SE) = 0.348 (0.142), p = 0.014). Unwanted behavior was significantly associated with male gender (adjusted B coefficient (SE) = 0.664 (0.321), p = 0.039) and COVID-19 positive status (adjusted B coefficient (SE) = 9.736 (3.297), p = 0.003). Knowledge of COVID-19 (adjusted B coefficient (SE) = 0.069 (0.035), p = 0.048) and being married (adjusted B coefficient (SE) = 0.917 (0.462), p = 0.048) were the predictors of desirable behavior. Conclusion: Overall, the UTAR community had demonstrated a good level of knowledge and preventive behaviors, albeit with some areas for improvement.

Isolation and Quantification of Mimivirus-Like and Marseillevirus-Like Viruses from Soil Samples in An Aboriginal (Orang asli) Village in Peninsular Malaysia.

BACKGROUND: The giant amoebal viruses of Mimivirus and Marseillevirus are large DNA viruses and have been documented in water, soil, and sewage samples. The trend of discovering these giant amoebal viruses has been increasing throughout Asia with Japan, India, and Saudi Arabia being the latest countries to document the presence of these viruses. To date, there have been no reports of large amoebal viruses being isolated in South East Asia. OBJECTIVE: In this study, we aim to discover these viruses from soil samples in an aboriginal village (Serendah village) in Peninsular -Malaysia. METHOD AND RESULTS: We successfully detected and isolated both Mimivirus-like and Marseillevirus-like viruses using Acanthamoeba castellanii. Phylogeny analysis identified them as Mimivirus and Marseillevirus, respectively. CONCLUSION: The ubiquitous nature of both Mimivirus and Marseillevirus is further confirmed in our study as they are detected in higher quantity in soil that is near to water vicinities in an aboriginal village in Peninsular Malaysia. However, this study is limited by our inability to investigate the impact of Mimivirus and Marseillevirus on the aboriginal villagers. More studies on the potential impact of these viruses on human health, especially on the aborigines, are warranted.

Association between polymorphisms of interleukin-17A G197A and interleukin-17F A7488G and risk of colorectal cancer.

BACKGROUND: Interleukin (IL)-17A and IL-17F are inflammatory cytokines mainly produced by T helper 17 cells. IL-17A is known to be protumorigenic while IL-17F has a protective role in cancer. A number of studies have been conducted to determine the association between polymorphisms of IL-17AG197A (rs2275913) and IL-17FA7488G (rs763780) and risk of cancers. No studies have yet to be conducted to genotype the IL-17AG197A polymorphism in colorectal cancer (CRC). OBJECTIVE: To assess the association of IL-17AG197A and IL-17FA7488G polymorphisms with CRC risk. MATERIALS AND METHODS: We performed the genotyping by polymerase chain reaction-restriction fragment length polymorphism method on blood samples from 80 healthy individuals and paraffin-embedded tumor tissues from 70 CRC patients. RESULTS: Our study showed that IL-17A197AA genotype was significantly associated with an increased CRC risk with odds ratios of 6.08 (95% confidence interval [CI]: 2.25-16.42, P < 0.001) and 2.80 (95% CI: 1.23-6.35, P = 0.014), in comparison with GG and AG genotypes, respectively. However, IL-17FA7488G polymorphism was not significantly associated with CRC risk (P = 0.102). No significant association of IL-17AG197A and IL-17FA7488G polymorphisms with patient and tumor variables was found. CONCLUSION: This report from Malaysia shows the relationship of IL-17A197AA genotype with susceptibility to CRC.

Risk factors associated with abdominal obesity in suburban adolescents from a Malaysian district.

INTRODUCTION: We aimed to determine the risk factors associated with abdominal obesity (AO) in suburban adolescents. METHODS: This cross-sectional study included adolescents aged 15-17 years from five randomly selected secondary schools in the Hulu Langat district of Selangor state, Malaysia. Waist circumference (WC) was measured at the midpoint between the lower margin of the last palpable rib and the top of the iliac crest. Information on sociodemographic data, dietary habits, physical activity levels and duration of sleep was obtained via interviewer-administered questionnaires. Participants' habitual food intake was determined using a 73-item Food Frequency Questionnaire. RESULTS: Among 832 participants, 56.0% were girls; 48.4% were Malay, 40.5% Chinese, 10.2% Indian and 0.8% of other ethnic groups. Median age and WC were 16 (interquartile range [IQR] 15-16) years and 67.9 (IQR 63.0-74.6) cm, respectively. Overall prevalence of AO (> 90th percentile on the WC chart) was 11.3%. A higher proportion (22.4%) of Indian adolescents were found to have AO compared with Malay and Chinese adolescents. Logistic regression analysis showed that female gender (adjusted odds ratio [OR] 7.064, 95% confidence interval [CI] 2.087-23.913; p = 0.002), Indian ethnicity (adjusted OR 10.164, 95% CI 2.182-47.346; p = 0.003), irregular meals (adjusted OR 3.193, 95% CI 1.043-9.774; p = 0.042) and increasing body mass index (BMI) (adjusted OR 2.867, 95% CI 2.216-3.710; p < 0.001) were significantly associated with AO. CONCLUSION: AO was common among Malaysian adolescents. Female gender, Indian ethnicity, irregular meals and increasing BMI were significant risk factors.

Generation of a MLL-AF9-specific stem cell model of acute monocytic leukemia.

Acute monocytic leukemia (AML-M5), a subtype of acute myeloid leukemia (AML), affects mostly young children and has poor prognosis. The mechanisms of treatment failure of AML-M5 are still unclear. In this study, we generated iPSC from THP-1 cells from a patient with AML-M5, using retroviruses encoding the pluripotency-associated genes (OCT3/4, SOX2, KLF4 and c-MYC). These AML-M5-derived iPSC showed features similar with those of human embryonic stem cells in terms of the morphology, gene expression, protein/antigen expression and differentiation capability. Parental-specific markers were down-regulated in these AML-M5-derived iPSCs. Expression of MLL-AF9 fusion gene (previously identified to be associated with pathogenesis of AML-M5) was observed in all iPSC clones as well as parental cells. We conclude that AML-M5-specific iPSC clones have been successfully developed. This disease model may provide a novel approach for future study of pathogenesis and therapeutic intervention of AML-M5.

Pteropine orthoreovirus infection among out-patients with acute upper respiratory tract infection in Malaysia.

This study aims to assess the incidence rate of Pteropine orthreovirus (PRV) infection in patients with acute upper respiratory tract infection (URTI) in a suburban setting in Malaysia, where bats are known to be present in the neighborhood. Using molecular detection of PRVs directly from oropharyngeal swabs, our study demonstrates that PRV is among one of the common causative agents of acute URTI with cough and sore throat as the commonest presenting clinical features. Phylogenetic analysis on partial major outer and inner capsid proteins shows that these PRV strains are closely related to Melaka and Kampar viruses previously isolated in Malaysia. Further study is required to determine the public health significance of PRV infection in Southeast Asia, especially in cases where co-infection with other pathogens may potentially lead to different clinical outcomes.

Prevalence of obesity and its associated risk factors among Chinese adults in a Malaysian suburban village.

INTRODUCTION: Obesity is a major modifiable risk factor associated with most chronic diseases. The aim of this study was to determine the prevalence of obesity, and its associated risk factors, among apparently healthy Chinese adults in a Malaysian suburban village. METHODS: This was a cross-sectional study conducted among the Chinese residents in Seri Kembangan New Village, Klang Valley, Selangor, Malaysia. Convenience sampling was used for the selection of participants. Body weight, height, waist and hip circumferences, and blood pressure were measured. Fasting venous plasma was drawn for the measurement of fasting glucose level and lipid profile. Data on sociodemographic factors, dietary habits, physical activity, perceived stress level and sleep duration were collected using interviewer-administered, pretested and validated questionnaires. RESULTS: Among the 258 Chinese residents (mean age 41.4 ± 10.0 years) recruited, the prevalence of obesity was 40%. The obese participants had significantly higher mean blood pressure, and triglyceride and fasting plasma glucose levels than the non-obese participants (p < 0.05). The obese participants also had a significantly lower high-density lipoprotein cholesterol level than the non-obese participants. Logistic regression analysis showed that drinking soy milk (adjusted odds ratio [OR] 0.447; 95% confidence interval [CI] 0.253-0.787; p < 0.05) and the perception that a balanced diet consists mainly of vegetables (adjusted OR 0.440; 95% CI 0.215-0.900; p < 0.05) were associated with a reduced risk of obesity. The risk of obesity was higher in younger participants (adjusted OR 2.714; 95% CI 1.225-6.011; p < 0.05). CONCLUSION: The prevalence of obesity was high among the apparently healthy suburban Chinese. Our findings suggest that soy milk consumption and the perception that a balanced diet consists mainly of vegetables are associated with a lower risk of developing obesity in this population.

Molecular alterations of Ras-Raf-mitogen-activated protein kinase and phosphatidylinositol 3-kinase-Akt signaling pathways in colorectal cancers from a tertiary hospital at Kuala Lumpur, Malaysia.

Molecular alterations in KRAS, BRAF, PIK3CA, and PTEN have been implicated in designing targeted therapy for colorectal cancer (CRC). The present study aimed to determine the status of these molecular alterations in Malaysian CRCs as such data are not available in the literature. We investigated the mutations of KRAS, BRAF, and PTEN, the gene amplification of PIK3CA, and the protein expression of PTEN and phosphatidylinositol 3-kinase (PI3K) catalytic subunit (p110α) by direct DNA sequencing, quantitative real-time PCR, and immunohistochemistry, respectively, in 49 CRC samples. The frequency of KRAS (codons 12, 13, and 61), BRAF (V600E), and PTEN mutations, and PIK3CA amplification was 25.0% (11/44), 2.3% (1/43), 0.0% (0/43), and 76.7% (33/43), respectively. Immunohistochemical staining demonstrated loss of PTEN protein in 54.5% (24/44) of CRCs and no significant difference in PI3K p110α expression between CRCs and the adjacent normal colonic mucosa (p = 0.380). PIK3CA amplification was not associated with PI3K p110α expression level, but associated with male cases (100% of male cases vs 56% of female cases harbored amplified PIK3CA, p = 0.002). PI3K p110α expression was significantly higher (p = 0.041) in poorly/moderately differentiated carcinoma compared with well-differentiated carcinoma. KRAS mutation, PIK3CA amplification, PTEN loss, and PI3K p110α expression did not correlate with Akt phosphorylation or Ki-67 expression. KRAS mutation, PIK3CA amplification, and PTEN loss were not mutually exclusive. This is the first report on CRC in Malaysia showing comparable frequency of KRAS mutation and PTEN loss, lower BRAF mutation rate, higher PIK3CA amplification frequency, and rare PTEN mutation, as compared with published reports.

HLA-A and breast cancer in West Peninsular Malaysia.

Breast cancer is the most common malignancy among females in Malaysia. Attempts have been made to investigate the association between breast cancer and human leukocyte antigen (HLA) types. However, data from those previous studies are highly variable. The aim of this study is to investigate the association between HLA-A types and clinicopathological factors in breast cancer. The frequencies of HLA-A type in 59 female patients with infiltrating ductal of the breast were determined by polymerase chain reaction method. HLA-A2/A30 and A2/A31 haplotype (5.1%; P = 0.045) as well as HLA-A30 (5.1%, P = 0.045) and A31 (6.8%; P = 0.020) allele were significant higher in the patients than controls (0%). HLA-A24 allele was negatively related to lymph node metastasis (r = -0.316; P = 0.021) whereas, A26 (r = -0.430; P = 0.001) and A36 (r = -0.430; P = 0.001) alleles were negatively correlated to distant metastasis in breast cancer. Negative correlations between HLA-A26/A36 (r = -0.430; P = 0.001), A2/A11 (r = -0.276; P = 0.044), A24/A34 (r = -0.430; P = 0.001) haplotypes and distant metastasis were identified. Interestingly, Her2 expression in breast carcinoma was negatively correlated to A11/24 haplotypes (r = -0.294; P = 0.034) but positively correlated to homozygous HLA-A24 (r = 0.396; P = 0.040). In conclusion, HLA-A2, -A30 and A31 were associated with breast cancer.

PIK3CA gene mutations in breast carcinoma in Malaysian patients.

Somatic mutations of phosphoinositide-3-kinase, catalytic, alpha; PIK3CA gene have been reported in several types of human cancers. The majority of the PIK3CA mutations map to the three "hot spots" - E542 K and E545 K in the helical (exon 9) and H1047R in the kinase (exon 20) domains of the p110alpha. These hot spot mutations lead to a gain of function in PI3 K signaling. We aimed to determine the frequency of PIK3CA mutations in the three most common Malaysian cancers. In this study, we assessed the genetic alterations in the PIK3CA gene in a series of 20 breast carcinomas, 24 colorectal carcinomas, 27 nasopharyngeal carcinomas (NPC), and 5 NPC cell lines. We performed mutation analysis of the PIK3CA gene by genomic polymerase chain reaction (PCR) and followed by DNA direct sequencing in exons 9 and 20. No mutations were detected in any of the 24 colorectal and 27 NPC samples, but one hot spot mutation located at exon 20 was found in a NPC cell line, SUNE1. Interestingly, PIK3CA somatic mutations were present in 6/20 (30%) breast carcinomas. Two of the six mutations, H1047R, have been reported previously as a hot spot mutation. Only one out of three hot spot mutations were identified in breast tumor samples. The remaining four mutations were novel. Our data showed that a higher incidence rate of PIK3CA mutations was present in Malaysian breast cancers as compared to colorectal and nasopharyngeal tumor tissues. Our findings also indicate that PIK3CA mutations play a pivotal role in activation of the PI3 K signaling pathway in breast cancer, and specific inhibitors of PIK3CA could be useful for breast cancer treatment in Malaysia.

Phenotyping of lymphocytes expressing regulatory and effector markers in infiltrating ductal carcinoma of the breast.

Dysfunction of the host immune system in cancer patients can be due to a number of reasons including suppression of tumour associated antigen reactive lymphocytes by regulatory T (Treg) cells. In this study, we used flow cytometry to determine the phenotype and relative abundance of the tumour infiltrating lymphocytes (TILs) from 47 enzymatically dissociated tumour specimens from patients with infiltrating ductal carcinoma (IDC) of the breast. The expression of both effector and regulatory markers on the TILs were determined by using a panel of monoclonal antibodies. Analysis revealed CD8(+) T cells (23.4+/-2.1%) were predominant in TILs, followed by CD4(+) T cells (12.6+/-1.7%) and CD56(+) natural killer cells (6.4+/-0.7%). The CD4(+)/CD8(+) ratio was 0.8+/-0.9%. Of the CD8(+) cells, there was a higher number (68.4+/-3.5%) that expressed the effector phenotype, namely, CD8(+)CD28(+) and about 46% of this subset expressed the activation marker, CD25. Thus, a lower number of infiltrating CD8(+) T cells (31.6+/-2.8%) expressed the marker for the suppressor phenotype, CD8(+)CD28(-). Of the CD4(+) T cells, 59.6+/-3.9% expressed the marker for the regulatory phenotype, CD4(+)CD25(+). About 43.6+/-3.8% CD4(+)CD25(+) subset co-expressed both the CD152 and FOXP3, the Treg-associated molecules. A positive correlation was found between the presence of CD4(+)CD25(+) subset and age (> or =50 years old) (r=0.51; p=0.045). However, no significant correlation between tumour stage and CD4(+)CD25(+) T cells was found. In addition, we also found that the CD4(+)CD25(-) subset correlated with the expression of the nuclear oestrogen receptor (ER)-alpha in the tumour cells (r=0.45; p=0.040). In conclusion, we detected the presence of cells expressing the markers for Tregs (CD4(+)CD25(+)) and suppressor (CD8(+)CD28(-)) in the tumour microenvironment. This is the first report of the relative abundance of Treg co-expressing CD152 and FOXP3 in breast carcinoma.