Displaying publications 1521 - 1540 of 4699 in total

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  1. Sivasothy Y, Ibrahim H, Paliany AS, Alias SA, Md Nor NR, Awang K
    Planta Med, 2013 Dec;79(18):1775-80.
    PMID: 24356874 DOI: 10.1055/s-0033-1351075
    The rhizomes of Alpinia pahangensis yielded a new bis-labdanic diterpene for which the name pahangensin C (1) was proposed along with twelve known analogues (2-13). The structure of 1 was elucidated via spectroscopic methods including 1D and 2D NMR techniques and LCMS-IT-TOF analysis. Compounds 2 and 12 were isolated for the first time from the genus Alpinia. This is the second occurrence of compounds 2 and 12 in the Zingiberaceae family. Selected analogues exhibited moderate to strong inhibitory activity against Staphylococcus aureus and Bacillus cereus.
    Matched MeSH terms: Diterpenes/isolation & purification*; Plant Extracts/isolation & purification*
  2. Ch'ng YS, Loh YC, Tan CS, Ahmad M, Asmawi MZ, Wan Omar WM, et al.
    Pharm Biol, 2017 Dec;55(1):2083-2094.
    PMID: 28832263 DOI: 10.1080/13880209.2017.1357735
    CONTEXT: Vernonia amygdalina Del. (VA) (Asteraceae) is commonly used to treat hypertension in Malaysia.

    OBJECTIVE: This study investigates the vasorelaxant mechanism of VA ethanol extract (VAE) and analyzes its tri-step FTIR spectroscopy fingerprint.

    MATERIALS AND METHODS: Dried VA leaves were extracted with ethanol through maceration and concentrated using rotary evaporator before freeze-dried. The vasorelaxant activity and the underlying mechanisms of VAE using the cumulative concentration (0.01-2.55 mg/mL at 20-min intervals) were evaluated on aortic rings isolated from Sprague Dawley rats in the presence of antagonists.

    RESULTS: The tri-step FTIR spectroscopy showed that VAE contains alkaloids, flavonoids, and saponins. VAE caused the relaxation of pre-contracted aortic rings in the presence and absence of endothelium with EC50 of 0.057 ± 0.006 and 0.430 ± 0.196 mg/mL, respectively. In the presence of Nω-nitro-l-arginine methyl ester (EC50 0.971 ± 0.459 mg/mL), methylene blue (EC50 1.203 ± 0.426 mg/mL), indomethacin (EC50 2.128 ± 1.218 mg/mL), atropine (EC50 0.470 ± 0.325 mg/mL), and propranolol (EC50 0.314 ± 0.032 mg/mL), relaxation stimulated by VAE was significantly reduced. VAE acted on potassium channels, with its vasorelaxation effects significantly reduced by tetraethylammonium, 4-aminopyridine, barium chloride, and glibenclamide (EC50 0.548 ± 0.184, 0.158 ± 0.012, 0.847 ± 0.342, and 0.304 ± 0.075 mg/mL, respectively). VAE was also found to be active in reducing Ca2+ released from the sarcoplasmic reticulum and blocking calcium channels.

    CONCLUSIONS: The vasorelaxation effect of VAE involves upregulation of NO/cGMP and PGI2 signalling pathways, and modulation of calcium/potassium channels, and muscarinic and β2-adrenergic receptor levels.

    Matched MeSH terms: Plant Extracts/isolation & purification; Vasodilator Agents/isolation & purification
  3. Latifah I, Abdul Halim A, Rahmat MS, Nadia MF, Ubil ZE, Asmah H, et al.
    Malays J Pathol, 2017 08;39(2):161-166.
    PMID: 28866698 MyJurnal
    BACKGROUND: A study was conducted to confirm the status of rats as the carrier of pathogenic leptospira in Kuala Lumpur, Malaysia.

    METHOD: A total of 140 urine samples were collected from trapped rats. These samples were cultured in EMJH enriched media and 18 of these samples (12.9%) were found to be positive when observed under x40 by dark field microscope. Genomic DNA was extracted from all the 18 native isolates for PCR.

    RESULT: All the 18 isolates generated the expected 786 base pair band when the set of primers known to amplify LipL32 gene were utilized. These results showed that the primers were suitable to be used for the identification of pathogenic leptospira from the 18 rat samples.

    CONCLUSION: The sequencing of the PCR products and BLAST analysis performed on each representative isolates confirmed the pathogenic status of all these native isolates as the LipL32 gene was detected in all the Leptospira isolates. This indicates that the rats are carriers of the pathogenic leptospira in the study area, and therefore are of public health importance.

    Matched MeSH terms: Bacterial Outer Membrane Proteins/isolation & purification; Lipoproteins/isolation & purification
  4. Kumaran S, Pandurangan AK, Shenbhagaraman R, Esa NM
    Int J Med Mushrooms, 2017;19(8):675-684.
    PMID: 29199567 DOI: 10.1615/IntJMedMushrooms.2017021274
    The growth and lectin production of Ganoderma applanatum, a white rot fungus, was optimized in broth cultures. The fungus was found to have a higher growth rate and higher lectin activity when grown in a medium adjusted to pH 6.5 at 26°C under stationary conditions. Expression of lectin activity started in 5-day-old mycelial culture; maximum activity was expressed after the 15th day of incubation. Among the various carbon and nitrogen sources tested, the carbon source sucrose and the nitrogen source yeast extract support maximum growth and lectin production. Lectin from G. applanatum was purified by ammonium sulfate precipitation and ion exchange chromatography. The purified fraction revealed a single band with a molecular weight of 35.0 kDa. Moreover, carbohydrates such as mannitol, glucose, sucrose, maltose, mannose, galactose, sorbose, and fructose were found to inhibit the hemagglutinating activity of the lectin. The purified lectins from G. applanatum contain cytotoxic and proapoptotic activities against HT-29 colon adenocarcinoma cells.
    Matched MeSH terms: Antineoplastic Agents/isolation & purification*; Lectins/isolation & purification
  5. Henry Sum MS, Yee SF, Eng L, Poili E, Lamdin J
    Biomed Res Int, 2017;2017:3608042.
    PMID: 29201901 DOI: 10.1155/2017/3608042
    Rice tungro disease (RTD) is one of the most destructive diseases of rice in South and Southeast Asia. RTD is routinely detected based on visual observation of the plant. However, it is not always easy to identify the disease in the field as it is often confused with other diseases or physiological disorders. Here we report the development of two serological based assays for ease of detection of RTD. In this study we had developed and optimized an indirect ELISA and dot-blot assay for detection of RTD. The efficiency of both assays was evaluated by comparing the specificity and sensitivity of the assays to PCR assay using established primer sets. The indirect ELISA showed 97.5% and 96.6%, while the dot-blot assay showed 97.5% and 86.4% sensitivity and specificity, respectively, when compared to established PCR method. The high sensitivity and specificity of the two assays merit the use of both assays as alternative methods to diagnose RTD. Furthermore, the dot-blot assay is a simple, robust, and rapid diagnostic assay that is suitable for field test for it does not require any specialized equipment. This is a great advantage for diagnosing RTD in paddy fields, especially in the rural areas.
    Matched MeSH terms: RNA, Viral/isolation & purification; Waikavirus/isolation & purification*
  6. Chew YL, Mahadi AM, Wong KM, Goh JK
    BMC Complement Altern Med, 2018 Feb 20;18(1):70.
    PMID: 29463252 DOI: 10.1186/s12906-018-2137-5
    BACKGROUND: Bauhinia kockiana originates from Peninsular Malaysia and it is grown as a garden ornamental plant. Our previous study reported that this plant exhibited fairly strong antioxidant and antimicrobial activities. This paper focused on the assessment of the antibacterial activity of B. kockiana towards methicillin-resistance Staphylococcus aureus (MRSA), to purify and to identify the antibacterial compounds, and to determine the mechanism of antibacterial activity.

    METHODS: Antibacterial activity of B. kockiana flower was evaluated qualitatively and quantitatively using disc diffusion assay and microbroth dilution method. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of extracts were examined. Phytochemical analysis was performed to determine the classes of phytochemicals in the extracts. Bioactivity guided isolation was employed to purify the antibacterial agents and identified via various spectroscopy methods. Scanning electron microscopy (SEM) technique was used to evaluate the antibacterial mechanism of extract and compounds isolated.

    RESULTS: B. kockiana flower was found to exhibit fairly strong antibacterial activity towards both strains of MRSA bacteria used, MIC varies from 62.5-250 μg/mL. Tannins and flavonoids have been detected in the phytochemical analysis. Gallic acid and its ester derivatives purified from ethyl acetate extract could inhibit MRSA at 250-500 μg/mL. SEM revealed that the cells have undergone plasmolysis upon treatment with the extract and compounds.

    CONCLUSION: Tannins and polyphenols are the antibacterial components towards MRSA in B. kockiana. Massive leakage of the cell content observed in treated cells showed that the phytochemicals have changed the properties of the cell membranes. Amphiphilic nature of the compounds exhibited the antibacterial activity towards MRSA via three stages: (1) cell membrane attachment; (2) cell membrane fluidity modification; and (3) cell membrane structure disruption.

    Matched MeSH terms: Anti-Bacterial Agents/isolation & purification; Plant Extracts/isolation & purification
  7. Chong SE, Mohamad Zaini RH, Suraiya S, Lee KT, Lim JA
    Malar J, 2017 01 03;16(1):2.
    PMID: 28049485 DOI: 10.1186/s12936-016-1666-y
    BACKGROUND: Dengue and malaria are two common, mosquito-borne infections, which may lead to mortality if not managed properly. Concurrent infections of dengue and malaria are rare due to the different habitats of its vectors and activities of different carrier mosquitoes. The first case reported was in 2005. Since then, several concurrent infections have been reported between the dengue virus (DENV) and the malaria protozoans, Plasmodium falciparum and Plasmodium vivax. Symptoms of each infection may be masked by a simultaneous second infection, resulting in late treatment and severe complications. Plasmodium knowlesi is also a common cause of malaria in Malaysia with one of the highest rates of mortality. This report is one of the earliest in literature of concomitant infection between DENV and P. knowlesi in which a delay in diagnosis had placed a patient in a life-threatening situation.

    CASE PRESENTATION: A 59-year old man staying near the Belum-Temengor rainforest at the Malaysia-Thailand border was admitted with fever for 6 days, with respiratory distress. His non-structural protein 1 antigen and Anti-DENV Immunoglobulin M tests were positive. He was treated for severe dengue with compensated shock. Treating the dengue had so distracted the clinicians that a blood film for the malaria parasite was not done. Despite aggressive supportive treatment in the intensive care unit (ICU), the patient had unresolved acidosis as well as multi-organ failure involving respiratory, renal, liver, and haematological systems. It was due to the presentation of shivering in the ICU, that a blood film was done on the second day that revealed the presence of P. knowlesi with a parasite count of 520,000/μL. The patient was subsequently treated with artesunate-doxycycline and made a good recovery after nine days in ICU.

    CONCLUSIONS: This case contributes to the body of literature on co-infection between DENV and P. knowlesi and highlights the clinical consequences, which can be severe. Awareness should be raised among health-care workers on the possibility of dengue-malaria co-infection in this region. Further research is required to determine the real incidence and risk of co-infection in order to improve the management of acute febrile illness.

    Matched MeSH terms: Dengue Virus/isolation & purification*; Plasmodium knowlesi/isolation & purification*
  8. Zaw MT, Emran NA, Ibrahim MY, Suleiman M, Awang Mohd TA, Yusuff AS, et al.
    J Microbiol Immunol Infect, 2019 Aug;52(4):563-570.
    PMID: 29428381 DOI: 10.1016/j.jmii.2018.01.003
    BACKGROUND: Cholera is an important health problem in Sabah, a Malaysian state in northern Borneo; however, Vibrio cholerae in Sabah have never been characterized. Since 2002, serogroup O1 strains having the traits of both classical and El Tor biotype, designated as atypical El Tor biotype, have been increasingly reported as the cause of cholera worldwide. These variants are believed to produce clinically more severe disease like classical strains.

    PURPOSE: The purpose of this study is to investigate the genetic diversity of V.cholerae in Sabah and whether V.cholerae in Sabah belong to atypical El Tor biotype.

    METHODS: ERIC-PCR, a DNA fingerprinting method for bacterial pathogens based on the enterobacterial repetitive intergenic consensus sequence, was used to study the genetic diversity of 65 clinical V.cholerae O1 isolates from 3 districts (Kudat, Beluran, Sandakan) in Sabah and one environmental isolate from coastal sea water in Kudat district. In addition, we studied the biotype-specific genetic traits in these isolates to establish their biotype.

    RESULTS: Different fingerprint patterns were seen in isolates from these three districts but one of the patterns was seen in more than one district. Clinical isolates and environmental isolate have different patterns. In addition, Sabah isolates harbor genetic traits specific to both classical biotype (ctxB-1, rstRCla) and El Tor biotype (rstRET, rstC, tcpAET, rtxC, VC2346).

    CONCLUSION: This study revealed that V.cholerae in Sabah were genetically diverse and were atypical El Tor strains. Fingerprint patterns of these isolates will be useful in tracing the origin of this pathogen in the future.

    Matched MeSH terms: DNA, Bacterial/isolation & purification; Vibrio cholerae O1/isolation & purification
  9. Chan YS, Cheah YH, Chong PZ, Khor HL, Teh WS, Khoo KS, et al.
    Pak J Pharm Sci, 2018 Jan;31(1):119-127.
    PMID: 29348093
    This study was conducted to investigate the antifungal potential and cytotoxicity of selected medicinal plants from Malaysia. The extracts from the stem of Cissus quadrangularis and the leaves of Asplenium nidus, Pereskia bleo, Persicaria odorata and Sauropus androgynus were assayed against six fungi using p-iodonitrotetrazolium-based on colorimetric broth microdilution method. All the plant extracts were found to be fungicidal against at least one type of fungus. The strongest fungicidal activity (minimum fungicidal concentration=0.16 mg/mL) were exhibited by the hexane extract of C. quadrangularis, the hexane, chloroform, ethanol and methanol extracts of P. bleo, the hexane and ethyl acetate extracts of P. odorata, and the water extract of A. nidus. In terms of cytotoxicity on the African monkey kidney epithelial (Vero) cells, the chloroform extract of P. odorata produced the lowest 50% cytotoxic concentration (100.3 ± 4.2 μ g/mL). In contrast, none of the water extracts from the studied plants caused significant toxicity on the cells. The water extract of A. nidus warrants further investigation since it showed the strongest fungicidal activity and the highest total activity (179.22 L/g) against Issatchenkia orientalis, and did not cause any toxicity to the Vero cells.
    Matched MeSH terms: Antifungal Agents/isolation & purification; Plant Extracts/isolation & purification
  10. Mohd-Ilham I, Muhd-Syafi AB, Khairy-Shamel ST, Shatriah I
    Singapore Med J, 2020 Jun;61(6):312-319.
    PMID: 31598730 DOI: 10.11622/smedj.2019121
    INTRODUCTION: Limited data is available on paediatric orbital cellulitis in Asia. We aimed to describe demographic data, clinical presentation, predisposing factors, identified microorganisms, choice of antibiotics and management in children with orbital cellulitis treated in a tertiary care centre in Malaysia.

    METHODS: A retrospective review was performed on children with orbital cellulitis aged below 18 years who were admitted to Hospital Universiti Sains Malaysia, Kelantan, Malaysia, between January 2013 and December 2017.

    RESULTS: A total of 14 paediatric patients fulfilling the diagnostic criteria for orbital cellulitis were included. Their mean age was 6.5 ± 1.2 years. Boys were more likely to have orbital cellulitis than girls (71.4% vs. 28.6%). Involvement of both eyes was observed in 14.3% of the patients. Sinusitis (28.6%) and upper respiratory tract infection (21.4%) were the most common predisposing causes. Staphylococcus aureus (28.6%) was the leading pathogen. Longer duration of hospitalisation was observed in those infected with methicillin-resistant Staphylococcus aureus and Burkholderia pseudomallei. 10 (71.4%) patients were treated with a combination of two or three antibiotics. In this series, 42.9% had surgical interventions.

    CONCLUSION: Young boys were found to be more commonly affected by orbital cellulitis than young girls. Staphylococcus aureus was the most common isolated microorganism. Methicillin-resistant Staphylococcus aureus and Burkholderia pseudomallei caused severe infection. Sinusitis and upper respiratory tract infection were the most common predisposing factors. A majority of the children improved with medical treatment alone. Our findings are in slight disagreement with other published reports on paediatric orbital cellulitis, especially from the Asian region.

    Matched MeSH terms: Staphylococcus aureus/isolation & purification; Methicillin-Resistant Staphylococcus aureus/isolation & purification
  11. Rajeh MA, Zuraini Z, Sasidharan S, Latha LY, Amutha S
    Molecules, 2010 Aug 31;15(9):6008-18.
    PMID: 20877206 DOI: 10.3390/molecules15096008
    The antimicrobial activities of the methanolic extracts of Euphorbia hirta L leaves, flowers, stems and roots were evaluated against some medically important bacteria and yeast using the agar disc diffusion method. Four Gram positive (Staphylococcus aureus, Micrococcus sp., Bacillus subtilis and Bacillus thuringensis), four Gram negative (Escherichia coli, Klebsiella pneumonia, Salmonella typhi and P. mirabilis) and one yeast (Candida albicans) species were screened. Inhibition zones ranged between 16-29 mm. Leaves extract inhibited the growth of all tested microorganisms with large zones of inhibition, followed by that of flowers, which also inhibited all the bacteria except C. albicans. The most susceptible microbes to all extracts were S. aureus and Micrococcus sp. Root extract displayed larger inhibition zones against Gram positive bacteria than Gram negative bacteria and had larger inhibition zones compared to stem extract. The lowest MIC values were obtained with E. coli and C. albicans (3.12 mg/mL), followed by S. aureus (12.50 mg/mL) and P. mirabilis (50.00 mg/mL). All the other bacteria had MIC values of 100.00 mg/mL. Scanning Electron Microscopic (SEM) studies revealed that the cells exposed to leaf extract displayed a rough surface with multiple blends and invaginations which increased with increasing time of treatment, and cells exposed to leaf extract for 36 h showed the most damage, with abundant surface cracks which may be related to final cell collapse and loss of function. Time-kill assay of C. albicans indicated a primarily fungicidal effect at 1- and 2-fold MIC. E. hirta extracts had LC(50) values of 0.71, 0.66, 0.41 and 0.03 mg/mL for stems, leaves, roots and flowers, respectively against Artemia salina. Hence, these plants can be used to discover new bioactive natural products that may serve as leads in the development of new pharmaceuticals.
    Matched MeSH terms: Anti-Bacterial Agents/isolation & purification*; Antifungal Agents/isolation & purification*
  12. Ma A, Wang Y, Liu XL, Zhang HM, Eamsobhana P, Yong HS, et al.
    J Helminthol, 2019 Jan;93(1):26-32.
    PMID: 29144215 DOI: 10.1017/S0022149X17001080
    Human gnathostomiasis is an emerging food-borne parasitic disease caused by nematodes of the genus Gnathostoma. Currently, serological tests are commonly applied to support clinical diagnosis. In the present study, a simple and rapid filtration-based test, dot immune-gold filtration assay (DIGFA) was developed using a partially purified antigen of Gnathostoma third-stage larvae (L3). A total of 180 serum samples were tested to evaluate the diagnostic potential of DIGFA for gnathostomiasis. The diagnostic sensitivity and specificity were 96.7% (29/30) and 100% (25/25), respectively. The cross-reactivity with sera from other helminthiasis patients ranged from 0 to 4%, with an average of 1.6% (2/125). DIGFA using a partially purified L3 antigen was not only simple and rapid, but also more accurate than standard assays for the diagnosis of human gnathostomiasis. DIGFA may represent a promising tool for application in laboratories or in the field, without requiring any instrumentation.
    Matched MeSH terms: Antigens, Helminth/isolation & purification; Gnathostoma/isolation & purification*
  13. Bae N, Li L, Lödl M, Lubec G
    Proc Natl Acad Sci U S A, 2012 Oct 30;109(44):17920-4.
    PMID: 23071323 DOI: 10.1073/pnas.1209632109
    Protein profiling has revealed the presence of glacontryphan-M, a peptide toxin identified only in the sea snail genus Conus, in the wings of Hebomoia glaucippe (HG). The wings and body of HG were homogenized and the proteins were extracted and analyzed by 2D gel electrophoresis with subsequent in-gel digestion. Posttranslational protein modifications were detected and analyzed by nano-LC-MS/MS. An antibody was generated against glacontryphan-M, and protein extracts from the wings of HG samples from Malaysia, Indonesia, and the Philippines were tested by immunoblotting. Glacontryphan-M was unambiguously identified in the wings of HG containing the following posttranslational protein modifications: monoglutamylation at E55, methylation at E53, quinone modification at W61, cyanylation at C56, and amidation of the C terminus at G63. Immunoblotting revealed the presence of the toxin in the wings of HG from all origins, showing a single band for glacontryphan-M in HG samples from Malaysia and Philippines and a double band in HG samples from Indonesia. Intriguingly, sequence analysis indicated that the Conus glacontryphan is identical to that of HG. The toxin may function as a defense against diverse predators, including ants, mantes, spiders, lizards, green frogs, and birds.
    Matched MeSH terms: Mollusk Venoms/isolation & purification*; Peptides, Cyclic/isolation & purification*
  14. Tam YJ, Allaudin ZN, Lila MA, Bahaman AR, Tan JS, Rezaei MA
    BMC Biotechnol, 2012 Oct 05;12:70.
    PMID: 23039947 DOI: 10.1186/1472-6750-12-70
    BACKGROUND: Cell disruption strategies by high pressure homogenizer for the release of recombinant Hepatitis B surface antigen (HBsAg) from Pichia pastoris expression cells were optimized using response surface methodology (RSM) based on the central composite design (CCD). The factors studied include number of passes, biomass concentration and pulse pressure. Polynomial models were used to correlate the above mentioned factors to project the cell disruption capability and specific protein release of HBsAg from P. pastoris cells.

    RESULTS: The proposed cell disruption strategy consisted of a number of passes set at 20 times, biomass concentration of 7.70 g/L of dry cell weight (DCW) and pulse pressure at 1,029 bar. The optimized cell disruption strategy was shown to increase cell disruption efficiency by 2-fold and 4-fold for specific protein release of HBsAg when compared to glass bead method yielding 75.68% cell disruption rate (CDR) and HBsAg concentration of 29.20 mg/L respectively.

    CONCLUSIONS: The model equation generated from RSM on cell disruption of P. pastoris was found adequate to determine the significant factors and its interactions among the process variables and the optimum conditions in releasing HBsAg when validated against a glass bead cell disruption method. The findings from the study can open up a promising strategy for better recovery of HBsAg recombinant protein during downstream processing.

    Matched MeSH terms: Hepatitis B Surface Antigens/isolation & purification; Recombinant Proteins/isolation & purification
  15. Thong KL, Goh YL, Radu S, Noorzaleha S, Yasin R, Koh YT, et al.
    J Clin Microbiol, 2002 Jul;40(7):2498-503.
    PMID: 12089269
    The incidence of food-borne salmonellosis due to Salmonella enterica serotype Weltevreden is reported to be on the increase in Malaysia. The pulsed-field gel electrophoresis (PFGE) subtyping method was used to assess the extent of genetic diversity and clonality of Salmonella serotype Weltevreden strains from humans and the environment. PFGE of XbaI-digested chromosomal DNA from 95 strains of Salmonella serotype Weltevreden gave 39 distinct profiles with a wide range of Dice coefficients (0.27 to 1.00), indicating that PFGE is very discriminative and that multiple clones of Salmonella serotype Weltevreden exist among clinical and environmental isolates. Strains of one dominant pulsotype (pulsotype X1/X2) appeared to be endemic in this region, as they were consistently recovered from humans with salmonellosis between 1996 and 2001 and from raw vegetables. In addition, the sharing of similar PFGE profiles among isolates from humans, vegetables, and beef provides indirect evidence of the possible transmission of salmonellosis from contaminated raw vegetables and meat to humans. Furthermore, the recurrence of PFGE profile X21 among isolates found in samples of vegetables from one wet market indicated the persistence of this clone. The environment in the wet markets may represent a major source of cross-contamination of vegetables with Salmonella serotype Weltevreden. Antibiotic sensitivity tests showed that the clinical isolates of Salmonella serotype Weltevreden remained drug sensitive but that the vegetable isolates were resistant to at least two antibiotics. To the best of our knowledge, this is the first study to compare clinical and environmental isolates of Salmonella serotype Weltevreden in Malaysia.
    Matched MeSH terms: DNA, Bacterial/isolation & purification; Salmonella enterica/isolation & purification*
  16. Pennie RA, Malik AS, Wilcox L
    J Clin Microbiol, 1996 May;34(5):1275-6.
    PMID: 8727917
    A 6-year-old boy presented to a university hospital in Malaysia with infective endocarditis complicating cyanotic congenital heart disease. Blood cultures showed a gram-positive, aerobic, coryneform-like bacillus identified by the hospital laboratory as Corynebacterium xerosis, but a reference laboratory identified the organism as a toxigenic strain of Corynebacterium diphtheriae. The two laboratories concurred on all biochemical test results except for sucrose fermentation.
    Matched MeSH terms: Corynebacterium/isolation & purification; Corynebacterium diphtheriae/isolation & purification
  17. Thong KL, Passey M, Clegg A, Combs BG, Yassin RM, Pang T
    J Clin Microbiol, 1996 Apr;34(4):1029-33.
    PMID: 8815078
    Molecular characterization of a total of 52 human isolates of Salmonella typhi from Papua New Guinea was performed by using pulsed-field gel electrophoresis (PFGE) after digestion of chromosomal DNA with three restriction endonucleases, XbaI (5'-TCTAGA-3'), AvrII (5'-CCTAGG-3'), and SpeI (5'-ACTAGT-3'). Of the 52 isolates tested, 11 were obtained from patients with fatal typhoid fever and 41 were obtained from patients with nonfatal disease. The 52 isolates showed limited genetic diversity as evidenced by only three different PFGE patterns detected following digestion with XbaI (patterns X1 to X3; F [coefficient of similarity] = 0.86 to 1.0), four patterns detected following digestion with AvrII (patterns A1 to A4; F =0.78 to 1.0), and two patterns detected following digestion with SpeI (patterns S1 and S2; F = 0.97 to 1.0). Of the 52 isolates, 37 were phage typed, and all belonged to phage type D2. All 11 isolates obtained from patients with fatal typhoid fever were identical (F = 1.0) and possessed the PFGE pattern combination X1S1A1, whereas the 41 isolates from patients with nonfatal typhoid fever had various PFGE pattern combinations, the most common being X2S1A2 (39%), X1S1A1 (24%), and X1S1A2 (15%). Thus, all the isolates from patients with the fatal disease had the X1 and A1 patterns, whereas the majority of the isolates from patients with nonfatal typhoid fever possessed the X2 and A2 patterns. The data suggest that there is an association among strains of S. typhi between genotype, as assessed by PFGE patterns, and the capability to cause fatal illness. Analysis of blood and fecal isolates of S. typhi from the same patient also indicated that some genetic changes occur in vivo during the course of infection.
    Matched MeSH terms: DNA, Bacterial/isolation & purification; Salmonella typhi/isolation & purification*
  18. Thong KL, Puthucheary S, Yassin RM, Sudarmono P, Padmidewi M, Soewandojo E, et al.
    J Clin Microbiol, 1995 Jul;33(7):1938-41.
    PMID: 7665677
    Pulsed-field gel electrophoresis (PFGE) revealed that multiple genetic variants of Salmonella typhi are simultaneously present in Southeast Asia and are associated with sporadic cases of typhoid fever and occasional outbreaks. Comparative analysis of PFGE patterns also suggested that considerable genetic diversity exists among S. typhi strains and that some PFGE patterns are shared between isolates obtained from Malaysia, Indonesia, and Thailand, implying movement of these strains within these regions of Southeast Asia, where they are endemic.
    Matched MeSH terms: DNA, Bacterial/isolation & purification; Salmonella typhi/isolation & purification*
  19. Thong KL, Cheong YM, Puthucheary S, Koh CL, Pang T
    J Clin Microbiol, 1994 May;32(5):1135-41.
    PMID: 7914202
    Pulsed-field gel electrophoresis (PFGE) was used to compare and analyze 158 isolates of Salmonella typhi from five well-defined outbreaks of typhoid fever in Malaysia and also isolates involved in sporadic cases of typhoid fever occurring during the same period. Digestion of chromosomal DNAs from these S. typhi isolates with the restriction endonucleases XbaI (5'-TCTAGA-3'), SpeI (5'-ACTAGT-3'), and AvrII (5'-CCTAGG-3') and then PFGE produced restriction endonuclease analysis (REA) patterns consisting of 11 to 24 DNA fragments ranging in size from 20 to 630 kbp. Analysis of the REA patterns generated by PFGE after digestion with XbaI and SpeI indicated that the S. typhi isolates obtained from sporadic cases of infection were much more heterogeneous (at least 13 different REA patterns were detected; Dice coefficient, between 0.73 and 1.0) than those obtained during outbreaks of typhoid fever. The clonal nature and the close genetic identities of isolates from outbreaks in Alor Setar, Penang, Kota Kinabalu, Johor Bahru, and Kota Bahru were suggested by the fact that only a limited number of REA patterns, which mostly differed by only a single band, were detected (one to four patterns; Dice coefficient, between 0.82 and 1.0), although a different pattern was associated with each of these outbreaks. Comparison of REA patterns with ribotyping for 18 S. typhi isolates involved in sporadic cases of infection showed a good correlation, in that 72% of the isolates were in the same group. There was no clear correlation of phage types with a specific REA pattern. We conclude that PFGE of s. typhi chromosomal DNA digested with infrequently cutting restriction endonucleases is a useful method for comparing and differentiating S. typhi isolates for epidemiological purposes.
    Matched MeSH terms: DNA, Bacterial/isolation & purification; Salmonella typhi/isolation & purification*
  20. Fu Z, Hamid SB, Razak CN, Basri M, Salleh AB, Rahman RN
    Protein Expr Purif, 2003 Mar;28(1):63-8.
    PMID: 12651108
    Bacteriocin release proteins (BRPs) can be used for the release of heterologous proteins from the Escherichia coli cytoplasm into the culture medium. The gene for a highly thermostable alkaline protease was cloned from Bacillus stearothermophilus F1 by the polymerase chain reaction. The recombinant F1 protease was efficiently excreted into the culture medium using E. coli XL1-Blue harboring two vectors: pTrcHis bearing the protease gene and pJL3 containing the BRPs. Both vectors contain the E. coli lac promoter-operator system. In the presence of 40 microM IPTG, the recombinant F1 protease and the BRP were expressed and mature F1 protease was released into the culture medium. This opens the way for the large-scale production of this protease in E. coli. The recombinant enzyme was purified through a one-step heat treatment at 70 degrees C for 3h and this method purified the protease to near homogeneity. The purified enzyme showed a pH optimum of 9.0, temperature optimum of 80 degrees C, and was stable at 70 degrees C for 24h in the pH range from 8.0 to 10.0. The enzyme exhibited a high degree of thermostability with a half-life of 4 h at 85 degrees C, 25 min at 90 degrees C, and was inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride (PMSF).
    Matched MeSH terms: Recombinant Proteins/isolation & purification; Serine Endopeptidases/isolation & purification*
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