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Fulltext Ethanol extracts of Allium sp. regulate cyclooxygenase-2 and E-cadherin expression in gastric cancer MKN74 cell line and enhance doxorubicin toxicity

Korga A, Ostrowska M, Iwan M, Skierucha M, Józefczyk A, Pawłowski P, et al.

Food Nutr Res, 2019;63.
PMID: 31297043 DOI: 10.29219/fnr.v63.3449

Background: Gastric cancer (GC) remains one of the leading causes of cancer-related death. Its aetiology is multifactorial, but the major risk factor is a high in salt diet. During gastric carcinogenesis, cadherin-1 (CDH1) down-expression and cyclooxygenase 2 (COX2) overexpression may be observed. The intensity of these alterations contributes to the GC invasion, its metastases and poor prognosis. As the diet plays a significant role in the aetiology of GC, it is reasonable to include the nutritional chemoprevention agents. One of the plant genus demonstrating chemoprotective properties is Allium genus, which includes garlic. The relationship between CDH1 and COX2 in GC cells treated with Allium species extract has never been evaluated.
Methods: In this study, the MKN28 and MKN74 GC cell lines were treated with ethanol extracts of Allium angulosum L., Allium lusitanicum Lam., Allium sativum L. (from Malaysia and Poland), Allium tibeticum Rendle and Allium ursinum L. The cytotoxicity of the extracts and their influence on COX2 and CDH1 mRNA and protein expression were evaluated as well as their influence on doxorubicin's (DOX) efficacy - a drug that has been used in GC treatment.
Results: Among the tested species, ethanol extracts of A. sativum L. (Poland and Malaysia), A. tibeticum Rendle and A. ursinum L. influenced the levels of CDH1 and COX2, but only in the MKN74 cell line. Thus, it is possible that tumours with increased COX2 expression will be more susceptible to garlic treatment. Observed phenomenon was independent of Allium extract's toxicity. In comparison to DOX, tested extracts were more toxic. Moreover, A. sativum revealed synergistic effect with the drug.
Conclusion: In conclusion, the results indicate the potential application of Allium genus to GC chemoprevention and treatment support through CDH restoration and COX2 downregulation. This issue needs further investigations as it might be used in clinics.

Matched MeSH terms: RNA, Messenger
Fulltext Hepatitis C infection and detection of antibodies, RNA and genotypes among female healthcare workers in Baghdad

Waqar, A.K., Nik Shamsidah N.I., Nor Aini M.N., Waqar, Abd Alqahar Al –Kubaisy

JUMMEC, 2018;21(1):14-20.
MyJurnal

Background: Hepatitis C Virus (HCV) is a major public health problem worldwide. About 130- 200 million people
are infected with HCV worldwide leading to 500,000 deaths annually (WHO 2014). Healthcare workers (HCWs)
have played an important role in the transmission of HCV infection, either as victims or as sources of infection.
Objectives: To determine the prevalence of HCV, antibodies (Abs) RNA and genotypes among the female HCWs
in Baghdad and to identify whether HCWs were infective or only infected.
Subjects and Methods: A cross-sectional study involving 1001 women attending 17 health care centres in
Baghdad, Iraq, was carried out. Information on type and duration of their occupation was obtained. HCV Abs
(anti-HCV) were tested using a third generation enzyme immunoassay (EIA-3) and immunoblot assay (Lia
Tek-111). Molecular analysis using RT-PCR and DNA enzyme immunoassay (DEIA) for HCV-RNA and genotype
detections were carried out for 63 serum samples.
Results: Only 160/1001 (15.98%) were HCWs. Anti-HCV and HCV- RNA seroprevalence were significantly higher
(6.37%, p=0.0057, 88.83%, p= 0.011 respectively) among HCWs than non HCWs. HCWs were at a significantly
higher risk of exposure to HCV infection (OR=2.75, 95% C.I. =1.31-5.79). There was no significant association
between HCV genotypes and the HCWs. HCV-4 showed higher expression (62.5%) among HCWs.
Conclusion: Female HCWs were infective and infected with HCV, thus there is a need for medical equipment
to be sterilized and cleaned thoroughly.

Matched MeSH terms: RNA
Fulltext Particulate matter (PM2.5) as a potential SARS-CoV-2 carrier

Nor NSM, Yip CW, Ibrahim N, Jaafar MH, Rashid ZZ, Mustafa N, et al.

Sci Rep, 2021 01 28;11(1):2508.
PMID: 33510270 DOI: 10.1038/s41598-021-81935-9

The rapid spread of the SARS-CoV-2 in the COVID-19 pandemic had raised questions on the route of transmission of this disease. Initial understanding was that transmission originated from respiratory droplets from an infected host to a susceptible host. However, indirect contact transmission of viable virus by fomites and through aerosols has also been suggested. Herein, we report the involvement of fine indoor air particulates with a diameter of ≤ 2.5 µm (PM2.5) as the virus's transport agent. PM2.5 was collected over four weeks during 48-h measurement intervals in four separate hospital wards containing different infected clusters in a teaching hospital in Kuala Lumpur, Malaysia. Our results indicated the highest SARS-CoV-2 RNA on PM2.5 in the ward with number of occupants. We suggest a link between the virus-laden PM2.5 and the ward's design. Patients' symptoms and numbers influence the number of airborne SARS-CoV-2 RNA with PM2.5 in an enclosed environment.

Matched MeSH terms: RNA, Viral
Fulltext A High Amylose Wheat Diet Improves Gastrointestinal Health Parameters and Gut Microbiota in Male and Female Mice

Lim SM, Choo JM, Li H, O'Rielly R, Carragher J, Rogers GB, et al.

Foods, 2021 Jan 21;10(2).
PMID: 33494480 DOI: 10.3390/foods10020220

High amylose wheat (HAW) contains more resistant starch than standard amylose wheat (SAW) and may have beneficial effects on gastrointestinal health. However, it is currently unclear whether these effects differ according to the level of HAW included in the diet or between males and females. Male and female C57BL/6 mice (n = 8/group/sex) were fed SAW65 (65% SAW; control), HAW35 (35% HAW), HAW50 (50% HAW) or HAW65 (65% HAW) diet for eight weeks. Female but not male, mice consuming any amount of HAW exhibited accelerated gastric emptying compared to SAW65 group. In both sexes, relative colon weights were higher in the HAW65 group compared to SAW65 group and in females, relative weights of the small intestine and cecum were also higher in the HAW65 group. In females only, colonic expression of Pyy and Ocln mRNAs were higher in the HAW65 group compared to HAW35 and HAW50 groups. In both sexes, mice consuming higher amounts of HAW (HAW50 or HAW65) had increased fecal bacterial load and relative abundance of Bacteroidetes phylum and reduced relative abundance of Firmicutes compared to SAW65 group. These data are consistent with a beneficial impact of HAW on gastrointestinal health and indicate dose-dependent and sex-specific effects of HAW consumption.

Matched MeSH terms: RNA, Messenger
Fucoidan Ameliorates Renal Injury-Related Calcium-Phosphorus Metabolic Disorder and Bone Abnormality in the CKD-MBD Model Rats by Targeting FGF23-Klotho Signaling Axis

Liu BH, Chong FL, Yuan CC, Liu YL, Yang HM, Wang WW, et al.

Front Pharmacol, 2020;11:586725.
PMID: 33708111 DOI: 10.3389/fphar.2020.586725

Background: Recently, chronic kidney disease (CKD)-mineral and bone disorder (MBD) has become one of common complications occurring in CKD patients. Therefore, development of a new treatment for CKD-MBD is very important in the clinic. In China, Fucoidan (FPS), a natural compound of Laminaria japonica has been frequently used to improve renal dysfunction in CKD. However, it remains elusive whether FPS can ameliorate CKD-MBD. FGF23-Klotho signaling axis is reported to be useful for regulating mineral and bone metabolic disorder in CKD-MBD. This study thereby aimed to clarify therapeutic effects of FPS in the CKD-MBD model rats and its underlying mechanisms in vivo and in vitro, compared to Calcitriol (CTR). Methods: All male rats were divided into four groups: Sham, CKD-MBD, FPS and CTR. The CKD-MBD rat models were induced by adenine administration and uninephrectomy, and received either FPS or CTR or vehicle after induction of renal injury for 21 days. The changes in parameters related to renal dysfunction and renal tubulointerstitial damage, calcium-phosphorus metabolic disorder and bone lesion were analyzed, respectively. Furthermore, at sacrifice, the kidneys and bone were isolated for histomorphometry, immunohistochemistry and Western blot. In vitro, the murine NRK-52E cells were used to investigate regulative actions of FPS or CTR on FGF23-Klotho signaling axis, ERK1/2-SGK1-NHERF-1-NaPi-2a pathway and Klotho deficiency. Results: Using the modified CKD-MBD rat model and the cultured NRK-52E cells, we indicated that FPS and CTR alleviated renal dysfunction and renal tubulointerstitial damage, improved calcium-phosphorus metabolic disorder and bone lesion, and regulated FGF23-Klotho signaling axis and ERK1/2-SGK1-NHERF-1-NaPi-2a pathway in the kidney. In addition, using the shRNA-Klotho plasmid-transfected cells, we also detected, FPS accurately activated ERK1/2-SGK1-NHERF-1-NaPi-2a pathway through Klotho loss reversal. Conclusion: In this study, we emphatically demonstrated that FPS, a natural anti-renal dysfunction drug, similar to CTR, improves renal injury-related calcium-phosphorus metabolic disorder and bone abnormality in the CKD-MBD model rats. More importantly, we firstly found that beneficial effects in vivo and in vitro of FPS on phosphorus reabsorption are closely associated with regulation of FGF23-Klotho signaling axis and ERK1/2-SGK1-NHERF-1-NaPi-2a pathway in the kidney. This study provided pharmacological evidences that FPS directly contributes to the treatment of CKD-MBD.

Matched MeSH terms: RNA, Small Interfering
Muscle cellularity, growth performance and growth-related gene expression of juvenile climbing perch Anabas testudineus in response to different eggs incubation temperature

Ahammad AKS, Asaduzzaman M, Uddin Ahmed MB, Akter S, Islam MS, Haque MM, et al.

J Therm Biol, 2021 Feb;96:102830.
PMID: 33627269 DOI: 10.1016/j.jtherbio.2020.102830

Although indigenous climbing perch (Anabas testudineusis) is a highly valuable species, slow growth pattern during the culture period impeding its commercial success in aquaculture. In many fish species, it has been demonstrated that incubation temperature of eggs influenced the muscle development and growth rates, which persisted throughout the subsequent larval and juvenile phases. Therefore, this study aimed to investigate whether different incubation temperature of eggs prior to hatching can stimulate the muscle development, growth, and growth-related gene expression of the slow-growing indigenous species of climbing perch. The fertilized eggs of A. testudineus from an artificial breeding program were incubated under control temperature of 24 °C (IT24), 26 °C (IT26), 28 °C (IT28), and 30 °C (IT30) in 10L glass aquaria with four replicated units for each temperature treatment. After hatching, the larvae from each incubated temperature were separately reared at ambient temperature for 10 days in aquarium, 20 days in hapas, and the next 42 days in cages, totaling 72 days post-hatching (dph). The hatching rates were found significantly (P

Matched MeSH terms: RNA, Messenger
Fulltext Molecular detection of feline leukemia virus in clinically ill cats in Klang Valley, Malaysia

Mummoorthy K, Yasmin AR, Arshad SS, Omar AR, Nur-Fazila SH, Anand P, et al.

Vet World, 2021 Feb;14(2):405-409.
PMID: 33776305 DOI: 10.14202/vetworld.2021.405-409

Background and Aim: Feline leukemia virus (FeLV) is classified as Retroviridae gammaretrovirus. FeLV occurs worldwide, including Malaysia. Thus far, only one decade-old study on molecular characterization of Malaysian FeLV isolates exists, which resulted in a scarcity of updated information of current FeLV isolates circulating in Malaysia. This study was conducted to determine the status of FeLV in clinically ill cats and to study the molecular characterization and phylogenetic relatedness of the current isolates.
Materials and Methods: Convenience sampling was performed in 20 cats from the Gasing Veterinary Hospital in Selangor. Plasma and saliva samples were collected from 15 clinically ill cats and 5 healthy cats subjected to one-step reverse transcription-polymerase chain reaction with primers targeting a highly conserved gene of U3-LTR-gag.
Results: Two clinically ill cats' plasma and saliva samples tested positive for FeLV RNA. Partial nucleotide sequencing and phylogenetic analysis revealed that the current isolates were 94-99% homologous to the previous Malaysian and Japanese FeLV isolates.
Conclusion: Current FeLV isolates from this study displayed higher similarity with the previous Malaysian isolates, signifying that a similar FeLV strain circulated among the cat population in Selangor.

Matched MeSH terms: RNA
In Vivo Antitumor Activity of Folate-Conjugated Cholic Acid-Polyethylenimine Micelles for the Codelivery of Doxorubicin and siRNA to Colorectal Adenocarcinomas

Amjad MW, Amin MC, Katas H, Butt AM, Kesharwani P, Iyer AK

Mol Pharm, 2015 Dec 7;12(12):4247-58.
PMID: 26567518 DOI: 10.1021/acs.molpharmaceut.5b00827

Multidrug resistance poses a great challenge to cancer treatment. In order to improve the targeting and codelivery of small interfering RNA (siRNA) and doxorubicin, and to overcome multidrug resistance, we conjugated a cholic acid-polyethylenimine polymer with folic acid, forming CA-PEI-FA micelles. CA-PEI-FA exhibited a low critical micelle concentration (80 μM), small average particle size (150 nm), and positive zeta potential (+ 12 mV). They showed high entrapment efficiency for doxorubicin (61.2 ± 1.7%, w/w), forming D-CA-PEI-FA, and for siRNA, forming D-CA-PEI-FA-S. X-ray photoelectron spectroscopic analysis revealed the presence of external FA on D-CA-PEI-FA micelles. About 25% doxorubicin was released within 24 h at pH 7.4, while more than 30% release was observed at pH 5. The presence of FA enhanced micelle antitumor activity. The D-CA-PEI-FA and D-CA-PEI-FA-S micelles inhibited tumor growth in vivo. No significant differences between their in vitro cytotoxic activities or their in vivo antitumor effects were observed, indicating that the siRNA coloading did not significantly increase the antitumor activity. Histological analysis revealed that tumor tissues from mice treated with D-CA-PEI-FA or D-CA-PEI-FA-S showed the lowest cancer cell density and the highest levels of apoptosis and necrosis. Similarly, the livers of these mice exhibited the lowest level of dihydropyrimidine dehydrogenase among all treated groups. The lowest serum vascular endothelial growth factor level (VEGF) (24.4 pg/mL) was observed in mice treated with D-CA-PEI-FA-S micelles using siRNA targeting VEGF. These findings indicated that the developed CA-PEI-FA nanoconjugate has the potential to achieve targeted codelivery of drugs and siRNA.

Matched MeSH terms: RNA, Small Interfering
Fulltext Familial hypercholesterolaemia: an updated overall management

Noor Alicezah Mohd Kasim, Chua Yung An, Hapizah Nawawi

Journal of Clinical and Health Sciences, 2020;5(2):19-38.
MyJurnal

Familial hypercholesterolaemia (FH), the commonest and serious but potentially treatable
form of inherited dyslipidaemias, is characterised by severely elevated plasma low-density
lipoprotein-cholesterol (LDL-C) level, which subsequently leads to premature coronary artery
disease (pCAD). Effectiveness of FH early detection and treatment is supported by the
outcome of several international cohort studies. Optimal FH management relies on
prescription of statins either alone or together with other lipid-lowering therapies (LLT).
Intensive lifestyle intervention is required in parallel with LLT, which should be commenced at
diagnosis in adults and childhood. Treatment with high intensity statin should be started as
soon as possible. Combination with ezetimibe and/or bile acid sequestrants is indicated if
target LDL-C is not achieved. For FH patients in the very-high risk category, if their LDL-C
targets are not achieved, despite being on maximally tolerated statin dose and ezetimibe,
proprotein convertase subtilisin/kexin type1 inhibitor (PCSK9i) is recommended. In statin
intolerance, ezetimibe alone, or in combination with PCSK9i may be considered. Clinical
evaluation of response to treatment and safety are recommended to be done about 4-6 weeks
following initiation of treatment. Homozygous FH (HoFH) patients should be treated with
maximally tolerated intensive LLT and, when available, with lipoprotein apheresis. This review
highlights the overall management, and optimal treatment combinations in FH in adults and
children, newer LLT including PCSK9i, microsomal transfer protein inhibitor, allele-specific
oligonucleotide to ApoB100 and PCSK9 mRNA. Family cascade screening and/or screening
of high-risk individuals, is the most cost-effective way of identifying FH cases and initiating
early and adequate LLT.

Matched MeSH terms: RNA, Messenger
Fulltext Milestone of polio Environmental Surveillance in Malaysia conjoining with the Polio Global Eradication Initiative

Kamal Haikal Mat Rabi, Amry Khursany Ismail, Mohd Samsul Samsuddin, Manisha Zauri Abdul Wahid, Zarina Mohd Zamawi, Ravindran Thayan

Malaysian Journal of Medicine and Health Sciences, 2019;15(106):78-78.
MyJurnal

Introduction: Poliomyelitis is an incapacitating and highly infectious disease which effect mostly young children. It is caused by one of the three serotypes of polioviruses (PV) and transmitted through faecal-oral route hence making the disease quite pertinent to the lower and middle class society or under-immunized population. This surveillance is one of the strategy included by WHO in the “Eradication, Integration and Certification: The Endgame Strategy 2019-2023” as a supplement to AFP surveillance by which it could be more sensitive to detect low circulation of WPV and circulating vaccine derived poliovirus (cVDPV). Methods: Routine collection and testing of representative environmental surveillance are carried out in the National Polio Laboratory. The specimens are collected from designated locations draining target populations at increased risk of poliovirus transmission using the grab method once a month and processed according to WHO standard protocol. Polioviruses were identified by real time reverse transcriptase polymerase chain reaction (rRT-PCR) for intratypic differentiation (ITD) and vaccine derived poliovirus (VDPV) whereas non-polio enteroviruses (NPEVs) were identified by PCR and sequencing. Results: From 2012 to 2019, results showed various isolation of PVs and NPEVs. A total of 12 sewage disposal plants located in urban highly populated areas in Kuala Lumpur (3), Selangor (5), Sabah (3 ) and Negeri Sembilan (1) were investigated. A total of 22 Sabin-like PVs were isolated consisting of 3 PV1, 8 PV2 and 11 PV3 thus indicated that in Malaysia even though PVs were existed in environment, but all of them were Sabin-Like viruses and no evidence of imported WPV or VDPV in the sampling sites. Conclusion: Even though Malaysia has been declared as WPV free country in 2000, Environmental Surveillance is very important and crucial in detecting the introduction and silent circulation of WPV and cVDPV before the virus reaches the community.

Matched MeSH terms: RNA-Directed DNA Polymerase
Fulltext Innovative models of HCV testing across four countries: Malaysia, Georgia, India and Myanmar – The HEAD-start project

Noor Annisa Darman, Muhammad Adib Abd Wahab, Wan Nur Illyana Wan Yusoff, Sasikala Siva, Xiao Hui Sem, Jessica Markby

Malaysian Journal of Medicine and Health Sciences, 2019;15(106):79-79.
MyJurnal

Introduction:Despite significant advancements in testing technologies and treatment, 80% of 80 million chronically hepatitis C virus (HCV) infected people remain undiagnosed. FIND in collaboration with Ministries of Health and local partners, initiated seven Unitaid funded Hepatitis C Elimination through Access to Diagnostics (HEAD-Start) projects in four countries (Georgia, India, Malaysia and Myanmar) to demonstrate innovative models of HCV care to inform programme scale-up nationally and globally. Here, we describe preliminary findings for patient retention in the HCV cascade for each project. Methods: Target populations and settings include people who inject drugs (PWID) in harm reduction sites (Manipur, Georgia, Myanmar), high risk patients in primary health care facilities (Malaysia), general population in polyclinics and district hospitals (Delhi) and people living with HIV (PLHIV) in antiretroviral therapy (ART) clinics (Punjab). All projects employ rapid diagnostic tests for HCV screening. HCV seropositive patients received confirmatory testing either on site (Georgia, Myanmar) or via centralized laboratory (Malaysia, Georgia, Delhi) or testing hubs (Punjab, Manipur). Results: In Malaysia, 4,613 patients were screened - 23% (1,042) are HCV seropositive, 57% (594) received RNA confirmatory testing and 11% (54) confirmed RNA-positive initiated treatment. In Delhi, 17,101 patients were screened – 2% (355) are seropositive, 92% (326) received confirmatory testing and 70% (180) initiated treatment. In Punjab, 17,507 patients were screened – 19% (3,368) are seroposi-tive, 94% (3,165) received confirmatory testing and 31% (828) initiated treatment. In Manipur, 3,845 patients were screened - 56% (2,180) are seropositive, 73% (1,591) received confirmatory testing and 67% (823) initiated treat-ment. In Myanmar, 199 patients were screened – 96% (191) are seropositive, 100% received confirmatory testing and 86% (151) initiated treatment. In Georgia (Arm 1) - 100% (619) received confirmatory testing and 76% (389) initiated treatment. Conclusion: The fully decentralised HCV testing model demonstrated higher numbers of patients progressing to each step in the care cascade.

Matched MeSH terms: RNA
Fulltext Safrana l Prevents Prostate Cancer Recurrence by Blocking the Re-activation of Quiescent Cancer Cells via Downregulation of S-Phase Kinase-Associated Protein 2

Jiang X, Li Y, Feng JL, Nik Nabil WN, Wu R, Lu Y, et al.

Front Cell Dev Biol, 2020;8:598620.
PMID: 33392189 DOI: 10.3389/fcell.2020.598620

The re-proliferation of quiescent cancer cells is considered to be the primary contributor to prostate cancer (Pca) recurrence and progression. In this study, we investigated the inhibitory effect of safranal, a monoterpene aldehyde isolated from Crocus sativus (saffron), on the re-proliferation of quiescent Pca cells in vitro and in vivo. The results showed that safranal efficiently blocked the re-activation of quiescent Pca cells by downregulating the G0/G1 cell cycle regulatory proteins CDK2, CDK4, CDK6, and phospho-Rb at Ser807/811 and elevating the levels of cyclin-dependent kinase inhibitors, p21 and p27. Further investigation on the underlying mechanisms revealed that safranal suppressed the mRNA and protein expression levels of Skp2, possibly through the deregulation of the transcriptional activity of two major transcriptional factors, E2F1 and NF-κB subunits. Moreover, safranal inhibited AKT phosphorylation at Ser473 and deregulated both canonical and non-canonical NF-κB signaling pathways. Safranal suppressed the tumor growth of quiescent Pca cell xenografts in vivo. Furthermore, safranal-treated tumor tissues exhibited a reduction in Skp2, E2F1, NF-κB p65, p-IκBα (Ser32), c-MYC, p-Rb (Ser807), CDK4, CDK6, and CDK2 and an elevation of p27 and p21 protein levels. Therefore, our findings demonstrate that safranal suppresses cell cycle re-entry of quiescent Pca cells in vitro and in vivo plausibly by repressing the transcriptional activity of two major transcriptional activators of Skp2, namely, E2F1 and NF-κB, through the downregulation of AKT phosphorylation and NF-κB signaling pathways, respectively.

Matched MeSH terms: RNA, Messenger
Fulltext Microbiome and colorectal cancer: an overview and local perspective

Rahman Jamal

Malaysian Journal of Medicine and Health Sciences, 2019;15(108):4-4.
MyJurnal

Colorectal cancer (CRC) is an important health problem that is on the rise globally, where it is the fourth most com-mon cause of deaths from cancer. CRC is now the 2nd commonest cancer in men and 3rd commonest in women in Malaysia. Diet, lifestyle, genetics and environmental interaction, together with underlying gut conditions such as inflammatory bowel disease have been reported to contribute to the disease. In addition, the gut microbiome has also been increasingly reported to be associated with CRC development, with dysbiosis of the commensal bacteria ob-served in CRC patients. Bacterial genera such as Bacteroides, Fusobacterium and Prevotella are more commonly de-tected in CRC patients compared to healthy individuals. Nevertheless, not much is known about the gut microbiome among Malaysians with different ethnicities. In Malaysia, the Chinese has the highest incidence of CRC, followed by Malays and Indians. The reason behind this difference may be contributed by the differences in the dietary intake that could modulate the gut microbiome and contribute towards the development of CRC. The current knowledge on this field still much depends on reports from individuals of American, European, Chinese, Brazilian and Japanese descendants in origin. The oncogenic potential of bacteria was suggested to include inflammation and the produc-tion of mutagenic toxin. A significant increase in certain intestinal microbiota including the genuses Enteroccus and Streptococcus spp. was detected in the advanced stage of colorectal adenoma. However, there are discrepancies in the previous studies, where some bacteria genera might be over-reported or underestimated. It is likely that the gut microbiome differs between populations. There is also no available data on the gut microbiome of the healthy individuals, colorectal adenoma (pre-cancerous) and colorectal cancer patients in the Malaysian population. Recent advancements in next generation sequencing allow faster and more accurate determination of microbial consortium in various niches of the human body and environment. In particular, sequencing of the 16S rRNA gene with specific primers have been reported to allow accurate determination of bacterial orders commonly found in the human gut as well as for those which are not expected in the digestive system. Recent developments in gut microbiome DNA ex-traction also contributed to the robustness of gut microbiome determination and analysis. All the above will contrib-ute towards an accurate and rapid cataloging process of the Malaysian gut microbiome and also enable comparison between healthy individuals, colorectal adenoma and CRC patients of the Malaysian population.

Matched MeSH terms: RNA, Ribosomal, 16S
Fulltext Annexin II Light Chain p11 Interacts With ENaC to Increase Functional Activity at the Membrane

Cheung TT, Ismail NAS, Moir R, Arora N, McDonald FJ, Condliffe SB

Front Physiol, 2019;10:7.
PMID: 30800070 DOI: 10.3389/fphys.2019.00007

The epithelial Na+ channel (ENaC) provides for Na+ absorption in various types of epithelia including the kidney, lung, and colon where ENaC is localized to the apical membrane to enable Na+ entry into the cell. The degree of Na+ entry via ENaC largely depends on the number of active channels localized to the cell membrane, and is tightly controlled by interactions with ubiquitin ligases, kinases, and G-proteins. While regulation of ENaC endocytosis has been well-studied, relatively little is understood of the proteins that govern ENaC exocytosis. We hypothesized that the annexin II light chain, p11, could participate in the transport of ENaC along the exocytic pathway. Our results demonstrate that all three ENaC channel subunits interacted with p11 in an in vitro binding assay. Furthermore, p11 was able to immunoprecipitate ENaC in epithelial cells. Quantitative mass spectrometry of affinity-purified ENaC-p11 complexes recovered several other trafficking proteins including HSP-90 and annexin A6. We also report that p11 exhibits a robust protein expression in cortical collecting duct epithelial cells. However, the expression of p11 in these cells was not influenced by either short-term or long-term exposure to aldosterone. To determine whether the p11 interaction affected ENaC function, we measured amiloride sensitive Na+ currents in Xenopus oocytes or mammalian epithelia co-expressing ENaC and p11 or a siRNA to p11. Results from these experiments showed that p11 significantly augmented ENaC current, whereas knockdown of p11 decreased current. Further, knockdown of p11 reduced ENaC cell surface population suggesting p11 promotes membrane insertion of ENaC. Overall, our findings reveal a novel protein interaction that controls the number of ENaC channels inserted at the membrane via the exocytic pathway.

Matched MeSH terms: RNA, Small Interfering
First Report of Soft Rot Caused by Pectobacterium carotovorum subsp. carotovorum on Pepper Fruits (Capsicum annuum) in Malaysia

Golkhandan E, Kamaruzaman S, Sariah M, Abidin MZZ, Nasehi A, Nazerian E

Plant Dis, 2013 Aug;97(8):1109.
PMID: 30722490 DOI: 10.1094/PDIS-01-13-0042-PDN

Symptoms of water-soaked lesions and soft rot were first observed in June 2011 on bell pepper fruits (Capsicum annuum cv. Annuum) in the two main regions of pepper production in Malaysia (Cameron Highlands and Johor State). Economic losses exceeded 40% in severely infected fields and greenhouses with the estimated disease incidence of 70%. In pepper fruits damaged by insects, sunscald, or other factors, symptoms initially appeared in the peduncle and calyx tissues and entire fruits were turned into watery masses within 2 to 6 days. Fruits infected in the field tended to collapse and hang on the plant. When the contents leaked out, the outer skin of the fruit dried and remained attached to the plant. Field-grown transplants and infected soil were identified as probable sources of inocula. A total of 50 attached fruits were collected from 10 pepper fields and greenhouses located in the two growing regions. Tissue from the margins of water-soaked lesions was surface-sterilized in 1% NaOCl for 2 min, rinsed in sterile water, dried, and plated onto nutrient agar (NA) and eosin methylene blue agar (EMB) media (3). A similar bacterium was isolated from all samples. After 2 days, white to creamy bacterial colonies on NA and emerald green colonies on EMB developed. Five independent strains were subjected to further biochemical, molecular, and pathogenicity tests. Bacterial strains were gram-negative, motile rods, grew at 37°C, were facultatively anaerobic, oxidase-negative, phosphatase-negative, and catalase-positive. They degraded pectate, were sensitive to erythromycin, did not utilize Keto-methyl glucoside, were indole production-negative, and reduced sugars from sucrose (3). Acid production was negative from sorbitol and arabitol, but positive from melibiose and citrate. PCR amplification of the pel gene by Y1 and Y2 primers produced a 434-bp fragment (2). Amplification of the intergenic transcribed spacer (ITS) region by G1 and L1 primers (4) gave two amplicons ca. 550 and 580 bp long. The expected amplicon was not produced with any of the strains using primers Br1f/L1r and Eca1f/Eca2r (1), whereas a 550-bp PCR product, typical of Pectobacterium carotovorum subsp. carotovorum, was obtained with primers EXPCCF and EXPCCR (1). Based on biochemical and molecular characteristics, and analysis of PCR-RFLP of 16S-ITS-23R rRNA genes using Rsa I enzyme (4), all five bacterial strains were identified as P. carotovorum subsp. carotovorum. BLAST analysis of the 16S rRNA sequence (GenBank Accession No KC189032) showed 100% identity to the 16S rRNA of P. carotovorum subsp. carotovorum strain PPC192. For pathogenicity tests, four mature pepper fruits of cv. Annuum were inoculated by injecting 10 μl of a bacterial suspension (108 CFU/ml) into pericarps and the fruits were incubated in a moist chamber at 80 to 90% relative humidity and 30°C. After 72 h, water-soaked lesions similar to those observed in the fields and greenhouses were observed and bacteria with the same characteristics were consistently reisolated, thereby fulfilling Koch's postulates. Symptoms were not observed on water-inoculated controls. References: (1) S. Baghaee-Ravari et al. Eur. J. Plant Pathol. 129:413, 2001. (2) A. Darraas et al. Appl. Environ. Microbiol. 60:1437, 1994. (3) N. W Schaad et al. Laboratory Guide for the Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society Press, St Paul, MN, 2001. (4) I. K. Toth et al. Appl. Environ. Microbiol. 67:4070, 2001.

Matched MeSH terms: RNA, Ribosomal, 16S
Fulltext Brain Beta-Catenin Signalling During Stress and Depression

Teo CH, Soga T, Parhar IS

Neurosignals, 2018 02 22;26(1):31-42.
PMID: 29490303 DOI: 10.1159/000487764

Beta-catenin is a protein with dual functions in the cell, playing a role in both adhesion between cells as well as gene transcription via the canonical Wnt signalling pathway. In the canonical Wnt signalling pathway, beta-catenin again plays multiple roles. In the embryonic stage, the regulation of beta-catenin levels activates genes that govern cell proliferation and differentiation. In an adult organism, beta-catenin continues to regulate the cell cycle - as a result over-expression of beta-catenin may lead to cancer. In the brain, dysfunctions in Wnt signalling related to beta-catenin levels may also cause various pathological conditions like Alzheimer's disease, Parkinson's disease, and depression. Beta-catenin can be influenced by stressful conditions and increases in glucocorticoid levels. In addition, beta-catenin can be regulated by neurotransmitters such as serotonin and dopamine. Fluctuations in beta-catenin in brain regions under duress have been associated with depressive-like behaviours. It is theorized that the change in behaviour can be attributed to the regulation of Dicer by beta-catenin. Dicer, a protein that produces micro-RNAs in the cell, is a target gene for beta-catenin. Amongst the micro-RNA that it produces are those involved in stress resilience. In this way, beta-catenin has taken its place in the well-studied biochemistry of stress and depression, and future research into this interesting protein may yet yield fruitful results in that field.

Matched MeSH terms: DEAD-box RNA Helicases
Intron retention is among six unreported AGL mutations identified in Malaysian GSD III patients

Abdullah IS, Teh SH, Khaidizar FD, Ngu LH, Keng WT, Yap S, et al.

Genes Genomics, 2019 08;41(8):885-893.
PMID: 31028654 DOI: 10.1007/s13258-019-00815-9

BACKGROUND: Glycogen storage disease type III is an autosomal recessive disorder that is caused by deficiencies of the glycogen debranching enzyme. Mutations within the AGL gene have been found to be heterogeneous, with some common mutations being reported in certain populations. The mutation spectrum of AGL gene in the multi-ethnic Malaysian population is still unknown.
OBJECTIVE: The present study seeks to determine the mutation spectrum of the AGL gene in Malaysian population.
METHODS: A total of eleven patients (eight Malay, two Chinese and one Bajau) were investigated. Genomic DNA was extracted and subsequently the AGL gene was amplified using specific primers and sequenced. Mutations found were screened in 150 healthy control samples either by restriction enzyme digestion assay or TaqMan® SNP Genotyping assay.
RESULTS: We identified six unreported mutations (c.1423+1G>T, c.2914_2915delAA, c.3814_3815delAG, c.4333T>G, c.4490G>A, c.4531_4534delTGTC) along with three previously reported mutations (c.99C>T, c.1783C>T, c.2681+1G>A). One of the six unreported mutation causes abnormal splicing and results in retention of intron 12 of the mature transcript, while another is a termination read-through. One of the reported mutation c.2681+1G>A was recurrently found in the Malay patients (n = 7 alleles; 31.8%).
CONCLUSION: The mutation spectrum of the AGL gene in Malaysian patients has shown considerable heterogeneity, and all unreported mutations were absent in all 150 healthy control samples tested.

Matched MeSH terms: RNA Splicing
Evaluation of wound healing activity of plumbagin in diabetic rats

Shao Y, Dang M, Lin Y, Xue F

Life Sci, 2019 Aug 15;231:116422.
PMID: 31059689 DOI: 10.1016/j.lfs.2019.04.048

This study was performed to evaluate the antidiabetic and wound healing activity of plumbagin in diabetic rats by macroscopical, biochemical, histological, immunohistochemical and molecular methods. Percentage of wound closure and contraction was delayed in diabetic rats when compared to non-diabetic group. There was significant reduction in period of epithelialization, collagen and protein content. Serum insulin level was significantly lowered together with increase in glucose level in diabetic rats. Lipid levels were increased significantly with concomitant decrease in HDL level. The mRNA levels of Nrf2, collagen-1, TGF-β and α-SMA were significantly lowered whereas Keap-1 levels were increased in diabetic rats. The level of lipid peroxides was increased while the levels of antioxidants were lowered significantly. ELISA results reveal upregulated levels of inflammatory markers. Western blot result shows upregulated levels of CD68 and CD163 proteins in wound area of diabetic rats. Histopathological observation revealed increased inflammatory cells infiltration in diabetic control. Immunofluorescent staining and immunohistochemical analysis also displayed delayed wound healing in diabetic groups. Diabetic rats treated with 10% and 20% plumbagin showed increased epithelialization, collagen deposition, increased serum insulin level and increased antioxidant status. Lipid peroxides and lipid levels were lowered significantly with increase in HDL level. Inflammatory markers were lowered, and growth factors expressions were increased markedly. Thus, the results of the study indicated that plumbagin administration could improve wound healing activity and could serve as a potent antidiabetic and anti-inflammatory agent.

Matched MeSH terms: RNA, Messenger
Combinatorial effects of genistein and sex-steroids on the level of cystic fibrosis transmembrane regulator (CFTR), adenylate cyclase (AC) and cAMP in the cervix of ovariectomised rats

Salleh N, Ismail N, Muniandy S, Korla PK, Giribabu N

Reprod Toxicol, 2015 Dec;58:194-202.
PMID: 26529183 DOI: 10.1016/j.reprotox.2015.10.017

The combinatorial effects of genistein and estrogen (E) or estrogen plus progesterone (E+P) on CFTR, AC and cAMP levels in cervix were investigated. Ovariectomised adult female rats received 50 or 100mg/kg/day genistein with E or E followed by E+P [E+(E+P)] for seven consecutive days. Cervixes were harvested and analyzed for CFTR mRNA levels by Real-time PCR. Distribution of AC and CFTR proteins in endocervix were observed by immunohistochemistry. Levels of cAMP were measured by enzyme-immunoassay. Molecular docking predicted interaction between genistein and AC. Our results indicate that levels of CFTR, AC and cAMP in cervix of rats receiving genistein plus E were higher than E-only treatment (p<0.05) while genistein plus [E+(E+P)] were higher than E+(E+P)-only treatment (p<0.05). In conclusions, increased levels of CFTR, AC and cAMP in cervix of E and E+(E+P)-treated rats by genistein could affect the cervical secretory function which could influence the female reproductive processes.

Matched MeSH terms: RNA, Messenger
Fulltext Data on PCR primer design for glucose 6-phosphate dehydrogenase gene and the effects of dietary carbohydrate levels on its expression in the liver of Malaysian mahseer (Tor tambroides)

Ishak SD, Razali SA, Kamarudin MS, Abol-Munafi AB

Data Brief, 2020 Aug;31:105916.
PMID: 32642522 DOI: 10.1016/j.dib.2020.105916

The enzyme glucose-6-phosphate dehydrogenase (G6PD) catalyses the metabolite glucose-6-phosphate in producing NADPH during the first phase of pentose-phosphate pathway thus provides reducing power to all cells for cellular growth, antioxidant defence, and biosynthetic reactions in all living organism. The deliberate inclusion of starch as carbohydrate source in commercial feed however may affect the G6PD hepatic activity in cultured fish. We designed a set of primers to target G6PD gene in the popular Malaysian aquaculture species, Tor tambroides. For this dataset, the molecular characteristics of obtained T. tambroides G6PD (TtG6PD) nucleotide sequence was analysed using multiple alignments and phylogenetic analyses of the deduced amino acids. The set of primers obtained were then used in a study to evaluate the effect of different dietary carbohydrate inclusion levels on the hepatic TtG6PD mRNA expression of the T. tambroides fingerlings. Four groups of fish were given a dietary treatment of 15%, 20%, 25% and 30% starch at the optimal inclusion level of 23.4% for 10 weeks. The TtG6PD mRNA transcripts were measured using real-time-PCR assays and its expression normalized against β-actin, which acts as the internal control gene. This article provides supportive data in relation between hepatic TtG6PD mRNA gene expression in T. tambroides and how it is influenced by its dietary carbohydrate intake. These data will also assist in further nutritional genomic studies of carbohydrate and energy utilization for all species in the mahseer family.

Matched MeSH terms: RNA, Messenger

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