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Studies on extraction of mannanase enzyme by Aspergillus terreus SUK-1 from fermented palm kernel cake

Rashid JI, Samat N, Yusoff WM

Pak J Biol Sci, 2013 Sep 15;16(18):933-8.
PMID: 24502150

Microbial mannanases have become biotechnologically important in industry but their application is limited due to high production cost. In presents study, the extraction of mannanase from fermented Palm Kernel Cake (PKC) in the Solid State Fermentation (SSF) was optimized. Local isolate of Aspergillus terreus SUK-1 was grown on PKC in (SSF) using column bioreactor. The optimum condition were achieved after two washes of fermented PKC by adding of 10% glycerol (v/v) soaked for 10 h at the room temperature with solvent to ratio, 1:5 (w/v).
Effect of some environmental parameters on biobutanol production by Clostridium acetobutylicum NCIMB 13357 in date fruit medium

Khamaiseh EI, Hamid AA, Yusoff WM, Kalil MS

Pak J Biol Sci, 2013 Oct 15;16(20):1145-51.
PMID: 24506014

Date fruit juice contains high concentration of simple sugars ranging from 65 to 75% (w/w) in dry form. In this study, the potential of date fruit juice as biobutanol fermentation medium by C. acetobutylicum was investigated. The fermentation process was carried out at initial pH of 5, 6 and 7, incubation temperature of 30, 35 and 40 degrees C for 72 hours. The date fruit concentrations tested were 10, 20, 30 and 40 g L(-1). Medium containing 30 g L(-1) of date fruit at 35 degrees C incubation temperature with initial medium pH 7.0 gave the highest concentration of solvents of 3.1, 0.1 and 1.1 g L(-1) butanol, ethanol and acetone respectively. The yield and productivity of biobutanol were 0.32 g g(-1) and 0.044 g L(-1)/h respectively, while for total ABE were 0.45 g g(-1) and 0.06 g L(-1) h, respectively.
Optimization of temperature, moisture content and inoculum size in solid state fermentation to enhance mannanase production by Aspergillus terreus SUK-1 using RSM

Rashid JI, Samat N, Mohtar W, Yusoff W

Pak J Biol Sci, 2011 May 01;14(9):533-9.
PMID: 22032082

Optimization of three parameters, temperature (25-35 degrees C), moisture content (40% (w/v)-60% (w/v) and inoculum sizes (5% (w/v)-15% (w/v) were investigated and optimized by Response Surface Methodology (RSM) for optimal mannanase production by Aspergillus terreus SUK-1. A second order polynomial equation was fitted and the optimum condition was established. The result showed that the moisture content was a critical factor in terms of its effect on mannanase. The optimum condition for mannanase production was predicted at 42.86% (w/v) initial moisture (31 C) temperature and 5.5% (w/v) inoculum size. The predicted optimal parameter were tested in the laboratory and the mannanase activity 45.12 IU mL-1 were recorded to be closed to the predicted value (44.80 IU mL-1). Under the optimized SSF condition (31 degrees C, 42.86% moisture content (w/v) and 5.5% inoculum size (w/v)), the maximum mannanase production was to prevail about 45.12 IU mL-1 compare to before optimized (30 degrees C, 50% moisture content (w/v) and 10% inoculum size (w/v)) was only 34.42 IU mL-1.
Fulltext Histopathological Alterations in Gills, Liver and Kidney of African Catfish (Clarias gariepinus, Burchell 1822) Exposed to Melaleuca cajuputi Extract

Hassan M, Melad AAN, Zakariah MI, Yusoff NAH

Trop Life Sci Res, 2023 Jun;34(2):177-196.
PMID: 38144386 DOI: 10.21315/tlsr2023.34.2.9

This study evaluated the histopathological changes in the gill, liver and kidney of African catfish (Clarias gariepinus) intoxicated with a sub-lethal dose of Melaleuca cajuputi leaves extract (MCLE) for 96 h. The acute toxicity test has been determined previously with a value of 96-h LC50 = 127 mg/L, hence the selection of sub-lethal ranges from 60 mg/L to 160 mg/L of MCLE. Degenerative alterations were prominent in all tested organs, particularly after exposure to a high concentration of MCLE. Gill exhibited haemorrhage, epithelial lifting, lamellar disorganisation, and necrosis after exposure to a high MCLE concentration. Alterations in the liver include congestion, hydropic degeneration, and vacuolation, whereas lesions in the kidney were pyknosis, vacuolation, hydropic degeneration, and tubular necrosis. The obtained data showed that the organs experienced severe changes proportional to the increase in MCLE concentration. In addition, fish exposed to higher concentrations than the LC50 value experienced irreversible lesions. The present study suggests that the use of MCLE below the LC50 is recommended to avoid severe alterations to organs, particularly in African catfish. This study demonstrated that the use of MCLE above the LC50 promotes severe damage to the gills, liver and kidney of African catfish. However, further investigations are needed to define the causing-mechanisms underlying these effects.
Fulltext Public perception on human exposure risk: A case study on endocrine disrupting compounds in the environment

Wee SY, Aris AZ, Yusoff FM, Praveena SM

Ecotoxicol Environ Saf, 2024 Jan 15;270:115830.
PMID: 38141339 DOI: 10.1016/j.ecoenv.2023.115830

Humans are exposed to environmental risks owing to the broad usage of endocrine disrupting compounds (EDCs). However, the subjective evaluation of risk levels and characteristics, as well as the variation in risk processing, have not been thoroughly examined. The objective was to understand the public's perception of the risk associated with human exposure to environmental EDCs and identify any variations in risk perception. In this pioneering study conducted within the distinctive social and cultural context of Malaysia, a developing nation, a quantitative analysis approach was employed to assess the subjective evaluation of risk levels and characteristics among the public while developing a risk perception model. Data gathered from surveys and questionnaires were analyzed to gather information on the public's perception of environmental and health issues pertaining to pesticides, hormones, plastics, medicines, and cosmetics. The analysis revealed that the majority of the public assessed the level of human exposure to environmental risks based on experiential processing, which was influenced by cognitive and affective variables. Interestingly, a higher proportion of individuals in the community had a low risk perception of environmental EDCs, surpassing the overall risk perception by 19.3%. Furthermore, the public showed significant awareness of environmental and health issues related to pesticides, hormones, and plastics but had a lesser inclination to acknowledge the vulnerability of humans to risks associated with medicines and cosmetics. These findings suggest that the public is likely to be exposed to environmental EDCs based on their current perceived risks, and that sociopsychological factors play a significant role in shaping perceptions and judgments. This understanding can inform the development of targeted risk management strategies and interventions to mitigate the potential harm caused by environmental EDCs.
WITHDRAWN: Rh genotypes among Malaysian blood donors

Musa RH, Hassan A, Ayob Y, Yusoff NM

Transfus Apher Sci, 2014 Dec 12.
PMID: 26144905 DOI: 10.1016/j.transci.2014.12.001

This article has been withdrawn at the request of the author and editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.
Fulltext In-Vitro and In-Silico Evaluations of Heterocyclic-Containing Diarylpentanoids as Bcl-2 Inhibitors Against LoVo Colorectal Cancer Cells

Leong SW, Chia SL, Abas F, Yusoff K

Molecules, 2020 Aug 26;25(17).
PMID: 32858795 DOI: 10.3390/molecules25173877

In the present study, we investigated the in-vitro anti-cancer potential of six diarylpentanoids against a panel of BRAF- and KRAS-mutated colorectal cancer cell lines including T84, SW620, LoVo, HT29, NCI-H508, RKO, and LS411N cells. Structure-activity relationship study suggested that the insertions of tetrahydro-4H-thiopyran-4-one and brominated phenyl moieties are essential for better cytotoxicity. Among the evaluated analogs, 2e has been identified as the lead compound due to its low IC50 values of approximately 1 µM across all cancer cell lines and high chemotherapeutic index of 7.1. Anti-proliferative studies on LoVo cells showed that 2e could inhibit cell proliferation and colony formations by inducing G2/M cell cycle arrest. Subsequent cell apoptosis assay confirmed that 2e is a Bcl-2 inhibitor that could induce intrinsic cell apoptosis by creating a cellular redox imbalance through its direct inhibition on the Bcl-2 protein. Further molecular docking studies revealed that the bromophenyl moieties of 2e could interact with the Bcl-2 surface pocket through hydrophobic interaction, while the tetrahydro-4H-thiopyran-4-one fragment could form additional Pi-sulfur and Pi-alkyl interactions in the same binding site. In all, the present results suggest that 2e could be a potent lead that deserves further modification and investigation in the development of a new Bcl-2 inhibitor.
Fulltext Reviews of the toxicity behavior of five potential engineered nanomaterials (ENMs) into the aquatic ecosystem

Jahan S, Yusoff IB, Alias YB, Bakar AFBA

Toxicol Rep, 2017;4:211-220.
PMID: 28959641 DOI: 10.1016/j.toxrep.2017.04.001

Presently, engineered nanomaterials (ENMs) are used in a wide variety of commercial applications, resulting in an uncontrolled introduction into the aquatic environment. The purpose of this review is to summarize the pathways and factors that controlling the transport and toxicity of five extensively used ENMs. These toxicological pathways are of great importance and need to be addressed for sustainable implications of ENMs without environmental liabilities. Here we discuss five potentially utilized ENMs with their possible toxicological risk factors to aquatic plants, vertebrates model and microbes. Moreover, the key effect of ENMs surface transformations by significant reaction with environmental objects such as dissolved natural organic matter (DOM) and the effect of ENMs surface coating and surface charge will also be debated. The transformations of ENMs are subsequently facing a major ecological transition that is expected to create a substantial toxicological effect towards the ecosystem. These transformations largely involve chemical and physical processes, which depend on the properties of both ENMs and the receiving medium. In this review article, the critical issues that controlling the transport and toxicity of ENMs are reviewed by exploiting the latest reports and future directions and targets are keenly discussed to minimize the pessimistic effects of ENMs.
Fulltext Delayed Diagnosis of Unilateral Absence of Pulmonary Artery Presenting with Recurrent Bronchopneumonia

Raja Shariff RE, Yusoff MR, Ibrahim KS, Kasim S

CASE (Phila), 2024 Mar;8(3Part A):103-108.
PMID: 38524981 DOI: 10.1016/j.case.2023.12.004

• Unilateral absence of the pulmonary artery (UAPA) is a rare congenital condition. • Patients with UAPA may present initially following recurrent bouts of pneumonia. • Echocardiography remains a useful tool for diagnosis in resource-limited settings.
Fulltext Hypoplastic Posterior Leaflet of the Tricuspid Valve-a Unique Cause of Right Heart Failure

Raja Shariff RE, Yusoff MR, Ibrahim KS, Kasim S

CASE (Phila), 2024 Mar;8(3Part A):157-161.
PMID: 38524993 DOI: 10.1016/j.case.2023.11.009

• TR can be due to either primary or secondary causes. • Primary TR due to congenital hypoplasia of leaflets is rare. • Multimodality imaging is key in identifying the cause of TR.
Fulltext Reduced Newcastle disease virus-induced oncolysis in a subpopulation of cisplatin-resistant MCF7 cells is associated with survivin stabilization

Jamal MH, Ch'ng WC, Yusoff K, Shafee N

Cancer Cell Int, 2012 Aug 01;12(1):35.
PMID: 22853623 DOI: 10.1186/1475-2867-12-35

BACKGROUND: Cisplatin resistance is a serious problem in cancer treatment. To overcome it, alternative approaches including virotherapy are being pursued. One of the candidates for anticancer virotherapy is the Newcastle disease virus (NDV). Even though NDV's oncolytic properties in various cancer cells have been widely reported, information regarding its effects on cisplatin resistant cancer cells is still limited. Therefore, we tested the oncolytic efficacy of a strain of NDV, designated as AF2240, in a cisplatin-resistant breast cancer cell line.
METHODS: Cisplatin-resistant cell line (MCF7-CR) was developed from the MCF7 human breast adenocarcinoma cell line by performing a seven-cyclic exposure to cisplatin. Following NDV infection, fluorescence-activated cell sorting (FACS) analysis and immunoblotting were used to measure cell viability and viral protein expression, respectively. Production of virus progeny was then assessed by using the plaque assay technique.
RESULTS: Infection of a mass population of the MCF7-CR with NDV resulted in 50% killing in the first 12 hours post-infection (hpi), comparable to the parental MCF7. From 12 hpi onwards, the remaining MCF7-CR became less susceptible to NDV killing. This reduced susceptibility led to increased viral protein synthesis and virus progeny production. The reduction was also associated with a prolonged cell survival via stabilization of the survivin protein.
CONCLUSIONS: Our findings showed for the first time, the involvement of survivin in the reduction of NDV-induced oncolysis in a subpopulation of cisplatin-resistant cells. This information will be important towards improving the efficacy of NDV as an anticancer agent in drug resistant cancers.
Fulltext The roles of emotional intelligence, neuroticism, and academic stress on the relationship between psychological distress and burnout in medical students

Yusoff MSB, Hadie SNH, Yasin MAM

BMC Med Educ, 2021 May 22;21(1):293.
PMID: 34022865 DOI: 10.1186/s12909-021-02733-5

BACKGROUND: Stress and burnout commonly threaten the mental health of medical students in Malaysia and elsewhere. This study aimed to explore the interrelations of psychological distress, emotional intelligence, personality traits, academic stress, and burnout among medical students.
METHODS: A cross-sectional study was conducted with 241 medical students. Validated questionnaires were administered to measure burnout, psychological distress, emotional intelligence, personality traits, and academic stress, respectively. A structural equation modelling analysis was performed by AMOS.
RESULTS: The results suggested a structural model with good fit indices, in which psychological distress and academic stress were noted to have direct and indirect effects on burnout. The burnout levels significantly increased with the rise of psychological distress and academic stress. Neuroticism was only found to have significant indirect effects on burnout, whereby burnout increased when neuroticism increased. Emotional intelligence had a significant direct effect on lowering burnout with the incremental increase of emotional intelligence, but it was significantly reduced by psychological distress and neuroticism.
CONCLUSION: This study showed significant effects that psychological distress, emotional intelligence, academic stress, and neuroticism have on burnout. Academic stress and neuroticism significantly increased psychological distress, leading to an increased burnout level, while emotional intelligence had a significant direct effect on reducing burnout; however, this relationship was compromised by psychological distress and neuroticism, leading to increased burnout. Several practical recommendations for medical educators, medical students, and medical schools are discussed.
Fulltext The Uprising of Mitochondrial DNA Biomarker in Cancer

Mohd Khair SZN, Abd Radzak SM, Mohamed Yusoff AA

Dis Markers, 2021;2021:7675269.
PMID: 34326906 DOI: 10.1155/2021/7675269

Cancer is a heterogeneous group of diseases, the progression of which demands an accumulation of genetic mutations and epigenetic alterations of the human nuclear genome or possibly in the mitochondrial genome as well. Despite modern diagnostic and therapeutic approaches to battle cancer, there are still serious concerns about the increase in death from cancer globally. Recently, a growing number of researchers have extensively focused on the burgeoning area of biomarkers development research, especially in noninvasive early cancer detection. Intergenomic cross talk has triggered researchers to expand their studies from nuclear genome-based cancer researches, shifting into the mitochondria-mediated associations with carcinogenesis. Thus, it leads to the discoveries of established and potential mitochondrial biomarkers with high specificity and sensitivity. The research field of mitochondrial DNA (mtDNA) biomarkers has the great potential to confer vast benefits for cancer therapeutics and patients in the future. This review seeks to summarize the comprehensive insights of nuclear genome cancer biomarkers and their usage in clinical practices, the intergenomic cross talk researches that linked mitochondrial dysfunction to carcinogenesis, and the current progress of mitochondrial cancer biomarker studies and development.
Fulltext Use of movement restrictions during an outbreak of COVID-19 in Selangor, Malaysia

Suleiman A, Ngadiman S, Ramly M, Yusoff AF, Yusof MP

Western Pac Surveill Response J, 2021 06 22;12(2):51-56.
PMID: 34540313 DOI: 10.5365/wpsar.2020.11.3.008

Objective: Various public health and social measures have been used during the COVID-19 outbreak, including lockdowns, contact-tracing, isolation and quarantine. The objective of this manuscript is to describe outbreaks of COVID-19 in Selangor, Malaysia, the public health strategies used and the observed impact of the measures on the epidemic curve.
Methods: Information on all confirmed COVID-19 cases in Selangor between 25 January and 28 April 2020 was obtained. Clusters were identified, and cases were disaggregated into linked, unlinked and imported cases. Epidemic curves were constructed, and the timing of movement control orders was compared with the numbers of cases reported.
Results: During the study period, 1395 confirmed COVID-19 cases were reported to the Selangor Health Department, of which 15.8% were imported, 79.5% were linked and 4.7% were unlinked cases. For two main clusters, the number of cases decreased after control measures were instituted, by contact-tracing followed by isolation and home quarantine for the first cluster (n = 126), and with the addition of the movement control order for the second, much larger cluster (n = 559).
Discussion: The findings suggest that appropriate, timely public health interventions and movement control measures have a synergistic effect on controlling COVID-19 outbreaks.
Fulltext Gelam Honey Inhibits the Production of Proinflammatory, Mediators NO, PGE(2), TNF-α, and IL-6 in Carrageenan-Induced Acute Paw Edema in Rats

Hussein SZ, Mohd Yusoff K, Makpol S, Mohd Yusof YA

Evid Based Complement Alternat Med, 2012;2012:109636.
PMID: 22919407 DOI: 10.1155/2012/109636

Natural honey is well known for its therapeutic value and has been used in traditional medicine of different cultures throughout the world. The aim of this study was to investigate the anti-inflammatory effect of Malaysian Gelam honey in inflammation-induced rats. Paw edema was induced by a subplantar injection of 1% carrageenan into the rat right hind paw. Rats were treated with the nonsteroidal anti-inflammatory drug (NSAID) Indomethacin (10 mg/kg, p.o.) or Gelam honey at different doses (1 or 2 g/kg, p.o.). The increase in footpad thickness was considered to be edema, which was measured using a dial caliper. Plasma and paw tissue were collected to analyze the production of inflammatory mediators, such as NO, PGE(2), TNF-α, and IL-6, as well as iNOS and COX-2. The results showed that Gelam honey could reduce edema in a dose-dependent fashion in inflamed rat paws, decrease the production of NO, PGE(2), TNF-α, and IL-6 in plasma, and suppress the expression of iNOS, COX-2, TNF-α, and IL-6 in paw tissue. Oral pretreatment of Gelam honey at 2 g/kg of body weight at two time points (1 and 7 days) showed a significantly decreased production of proinflammatory cytokines, which was similar to the effect of the anti-inflammatory drug Indomethacin (NSAID), both in plasma and tissue. Thus, our results suggest that Gelam honey has anti-inflammatory effects by reducing the rat paw edema size and inhibiting the production of proinflammatory mediators. Gelam honey is potentially useful for treating inflammatory conditions.
Fulltext Genetic diversity and phylogeographic patterns of the peacock jewel-damselfly, Rhinocypha fenestrella (Rambur, 1842)

Noorhidayah M, Azrizal-Wahid N, Low VL, Yusoff NR

PLoS One, 2024;19(4):e0301392.
PMID: 38578719 DOI: 10.1371/journal.pone.0301392

Despite is known to have widespread distribution and the most active species of the family Chlorocyphidae, the molecular data of Rhinocypha fenestrella (Rambur, 1842) are relatively scarce. The present study is the first that examined the genetic diversity and phylogeographic pattern of the peacock jewel-damselfly R. fenestrella by sequencing the cytochrome C oxidase I (cox1) and 16S rRNA gene regions from 147 individuals representing eight populations in Malaysia. A total of 26 and 10 unique haplotypes were revealed by the cox1 and 16S rRNA genes, respectively, and 32 haplotypes were recovered by the concatenated sequences of cox1+16S. Analyses indicated that haplotype AB2 was the most frequent and the most widespread haplotype in Malaysia while haplotype AB1 was suggested as the common ancestor haplotype of the R. fenestrella that may arose from the Negeri Sembilan as discovered from cox1+16S haplotype network analysis. Overall haplotype and nucleotide diversities of the concatenated sequences were Hd = 0.8937 and Pi = 0.0028, respectively, with great genetic differentiation (FST = 0.6387) and low gene flow (Nm = 0.14). Population from Pahang presented the highest genetic diversity (Hd = 0.8889, Pi = 0.0022, Nh = 9), whereas Kedah population demonstrated the lowest diversity (Hd = 0.2842, Pi = 0.0003, Nh = 4). The concatenated sequences of cox1+16S showed genetic divergence ranging from 0.09% to 0.97%, whereas the genetic divergence for cox1 and 16S rRNA genes were 0.16% to 1.63% and 0.01% to 0.75% respectively. This study provides for the first-time insights on the intraspecific genetic diversity, phylogeographic pattern and ancestral haplotype of Rhinocypha fenestrella. The understanding of molecular data especially phylogeographic pattern can enhance the knowledge about insect origin, their diversity, and capability to disperse in particular environments.
Fulltext The Characteristics and the Pharmacological Management of Cancer Pain and Its Effect on the Patients' Daily Activities and their Quality of Life: A Cross - Sectional study from Malaysia

K C B, Binti Mohd Yusoff Z, Alrasheedy AA, Othman S

J Clin Diagn Res, 2013 Jul;7(7):1408-13.
PMID: 23998077 DOI: 10.7860/JCDR/2013/5450.3148

CONTEXT: Pain is a major health care problem for the patients with cancer and one of the most frequent and disturbing cancer related symptoms.
AIM: To study the characteristics of pain in cancer patients and its pharmacological management by using a subjective self-assessment questionnaire and the World Health Organization (WHO)analgesic ladder for pain management.
SETTINGS AND DESIGNS: This study was conducted in the Oncology Wards of Penang Hospital, Penang, Malaysia. A questionnaire was developed to assess the pain characteristics and their effect on the patients' daily life activities and the information on the pharmacological management of the cancer pain. The cancer pain intensity was noted from the patients' medical database.
METHOD AND MATERIAL: By using the validated questionnaire, an observational, cross sectional study was conducted on the cancer patientswho were admitted in the oncology wards of Penang Hospital, Malaysia, for a period of 1 month.
STATISTICAL ANALYSIS: Descriptive statistics like mean, frequency and percentages were used for this study.
RESULTS AND CONCLUSION: A total of 42 patients out of 143, who fulfilled the criteria, were interviewed. The results showed mild pain in 66.7% (28) of the patients, moderate pain in 7.1% (3) and severe painin 26.2% (11). The normal daily life activities were affected by the pain in almost all the patients. Among the interviewed patients, sleep was affected in 88% (37) of the patients and the normal physical activity was affected in 92.9% (39) of the patients. Similarly, the pain decreased the appetite in 78.6% (33) of the patients, it affected the personal relationship in 35.7% (15), it affected the emotion in 71.5% (30) and it affected the visual activity in 33.6% (13) of the patients. Mild pain with distressing symptoms was not treated with any analgesic or adjuvant medications in 40.5% (17) of the patients. In contrast, all the patients with moderate and severe pain were treated with medications. Among them, 66.7% (2) of the patients with moderate pain and 90.9% (10) of the patients with severe pain were treated with analgesics as per the WHO analgesic ladder. The WHO analgesic ladder guide for pain management was followed in a majority of the cases, when analgesics were prescribed. However, there was inadequate treatment of the cancer pain in many patients with mild pain and, consequently, their quality of life was largely affected.
Surgical Management of Huge Ranula Resembling Double Tongue in Pediatric Patients: A Successful Treatment of Two Cases

Fauzi FA, Abdullah MF, Samsudin NA, Md Yusoff B

Cureus, 2024 Apr;16(4):e57884.
PMID: 38725744 DOI: 10.7759/cureus.57884

Ranula is a fluid collection in a pseudo cystic wall secondary to the damage of the sublingual salivary gland causing blockage of salivary flow, leading to the extravasation phenomena. The growth rate of ranula varies depending on its severity. Due to its tendency to recur, the gold standard management of ranula has yet to be decided. The authors described two cases of young girls with huge ranulas on the floor of the mouth (FOM) resembling double tongue, which caused pain and discomfort during mastication. Following surgical excision of the ranulas along with the affected sublingual glands, both cases demonstrated successful treatment outcomes with no recurrence observed during post-operative follow-up. These cases highlight the importance of surgical excision of ranulas and removal of affected sublingual glands to prevent recurrence.
Autophagy inhibition suppresses Newcastle disease virus-induced cell death by inhibiting viral replication in human breast cancer cells

Rozilah MI, Yusoff K, Chia SL, Ismail S

Virology, 2024 Feb;590:109957.
PMID: 38100982 DOI: 10.1016/j.virol.2023.109957

Newcastle disease virus (NDV) is an oncolytic virus which selectively replicates in cancer cells without harming normal cells. Autophagy is a cellular mechanism that breaks down unused cytoplasmic constituents into nutrients. In previous studies, autophagy enhanced NDV-induced oncolysis in lung cancer and glioma cells. However, the effect of autophagy inhibition on NDV-induced oncolysis in breast cancer cells remains unknown. This study aimed to examine the effect of autophagy inhibition on NDV-induced oncolysis in human breast cancer cells, MCF7. To inhibit autophagy, we knocked down the expression of the autophagy protein beclin-1 (BECN1) by short interfering RNA (siRNA). The cells were infected with the recombinant NDV strain AF2240 expressing green fluorescent protein. We found that NDV induced autophagy and knockdown of BECN1 significantly reduced the NDV-induced autophagy in MCF7 cells. Importantly, BECN1 knockdown significantly suppressed cell death by inhibiting viral replication, as observed at 24 h post infection. Overall, our data suggest that autophagy inhibition may not be a suitable strategy to enhance NDV oncolytic efficacy against breast cancer.
Purification and characterization of Nipah virus nucleocapsid protein produced in insect cells

Eshaghi M, Tan WS, Ong ST, Yusoff K

J Clin Microbiol, 2005 Jul;43(7):3172-7.
PMID: 16000431

The nucleocapsid (N) protein of Nipah virus (NiV) is a major constituent of the viral proteins which play a role in encapsidation, regulating the transcription and replication of the viral genome. To investigate the use of a fusion system to aid the purification of the recombinant N protein for structural studies and potential use as a diagnostic reagent, the NiV N gene was cloned into the pFastBacHT vector and the His-tagged fusion protein was expressed in Sf9 insect cells by recombinant baculovirus. Western blot analysis of the recombinant fusion protein with anti-NiV antibodies produced a band of approximately 62 kDa. A time course study showed that the highest level of expression was achieved after 3 days of incubation. Electron microscopic analysis of the NiV recombinant N fusion protein purified on a nickel-nitrilotriacetic acid resin column revealed different types of structures, including spherical, ring-like, and herringbone-like particles. The light-scattering measurements of the recombinant N protein also confirmed the polydispersity of the sample with hyrdrodynamic radii of small and large types. The optical density spectra of the purified recombinant fusion protein revealed a high A(260)/A(280) ratio, indicating the presence of nucleic acids. Western blotting and enzyme-linked immunosorbent assay results showed that the recombinant N protein exhibited the antigenic sites and conformation necessary for specific antigen-antibody recognition.

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