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  1. Phenotypic and genotypic characterization of two closely related subgroups of Candida rugosa in clinical specimens
    Tay ST, Tan HW, Na SL, Lim SL
    J Med Microbiol, 2011 Nov;60(Pt 11):1591-1597.
    PMID: 21700741 DOI: 10.1099/jmm.0.032854-0
    In this study, six clinical isolates (two from blood, two from urine and one each from a bronchoalveolar lavage and a vaginal swab) were identified as Candida rugosa based on carbohydrate assimilation profiles using API 20C AUX and ID32 C kits (bioMérieux). Sequence analysis of the D1/D2 domain of the yeasts differentiated the isolates into two subgroups, A and B (three isolates per subgroup), which were closely related (99.1-99.6 % nucleotide similarity) to C. rugosa strain ATCC 10571. Compared with the C. rugosa type strain, the intergenic transcribed spacer (ITS) nucleotide similarity for subgroup A was only 89.2 % (29 mismatches and one deletion) and for subgroup B was 93.7 % (20 mismatches). All isolates grew green colonies on Oxoid Chromogenic Candida Agar, with darker pigmentation observed for subgroup A. All isolates were able to grow at 25-42 °C but not at 45 °C. The isolates had identical enzymic profiles, as determined by API ZYM (bioMérieux) analysis, and produced proteinase. High amphotericin MICs (≥1 µg ml(-1)) were noted for two isolates from each subgroup. Dose-dependent susceptibility to fluconazole (MIC 32 µg ml(-1)) was noted in a blood isolate. The biofilms of the isolates demonstrated increased resistance to amphotericin and fluconazole. The greater ITS sequence variability of subgroup A isolates is in support of this yeast being recognized as a distinct species; however, further verification using more sophisticated molecular approaches is required. A sequence comparison study suggested the association of subgroup A with environmental sources and subgroup B with clinical sources. Accurate identification and antifungal susceptibility testing of C. rugosa are important in view of its decreased susceptibility to amphotericin and fluconazole. The ITS region has been shown to be a valuable region for differentiation of closely related subgroups of C. rugosa.
  2. Fulltext Molecular survey and sequence analysis of Anaplasma spp. in cattle and ticks in a Malaysian farm
    Tay ST, Koh FX, Kho KL, Ong BL
    Trop Biomed, 2014 Dec;31(4):769-76.
    PMID: 25776603 MyJurnal
    This study was conducted to determine the occurrence of Anaplasma spp. in the blood samples of cattle, goats, deer and ticks in a Malaysian farm. Using polymerase chain reaction (PCR) and sequencing approach, Anaplasma spp. was detected from 81(84.4%) of 96 cattle blood samples. All blood samples from 23 goats and 22 deer tested were negative. Based on the analysis of the Anaplasma partial 16S ribosomal RNA gene, four sequence types (genotypes 1 to 4) were identified in this study. Genotypes 1-3 showed high sequence similarity to those of Anaplasma platys/ Anaplasma phagocytophilum, whilst genotype 4 was identical to those of Anaplasma marginale/ Anaplasma centrale/ Anaplasma ovis. Anaplasma DNA was detected from six (5.5%) of 109 ticks which were identified as Rhipicephalus (formely known as Boophilus) microplus ticks collected from the cattle. This study reported for the first time the detection of four Anaplasma sequence types circulating in the cattle population in a farm in Malaysia. The detection of Anaplasma DNA in R. microplus ticks in this study provides evidence that the ticks are one of the potential vectors for transmission of anaplasmosis in the cattle.
  3. Fulltext Enzymatic profiling of clinical and environmental isolates of Burkholderia pseudomallei
    Liew SM, Tay ST, Wongratanacheewin S, Puthucheary SD
    Trop Biomed, 2012 Mar;29(1):160-8.
    PMID: 22543616 MyJurnal
    Melioidosis has been recognized as an important cause of sepsis in the tropics. The disease caused by an environmental saprophyte Burkholderia pseudomallei, affects mostly adults with underlying immunocompromised conditions. In this study, the enzymatic profiles of 91 clinical and 9 environmental isolates of B. pseudomallei were evaluated using the APIZYM system, in addition to assessment of protease, phospholipase C and sialidase activities using agar plate methods and other assays. The activity of 10 enzymes - alkaline phosphatase, esterase, esterase lipase, lipase, leucine arylamidase, valine arylamidase, cystine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase and N-acetyl-β-glucosaminidase were detected in >75% of the clinical isolates. The majority of B. pseudomallei isolates in this study exhibited protease and phospholipase activities. No sialidase activity was detected. Five Burkholderia thailandensis isolates had similar APIZYM profiles as B. pseudomallei clinical isolates except for the lower detection rate for N-acetyl-β-glucosaminidase. The subtle differences in the number of enzymes secreted and the levels of enzymatic activities of phenotypically identical clinical and environmental strains of B. pseudomallei give weight to the fact that the causative agent of melioidodis originates from the environment.
  4. Fulltext The integron prevalence of extended-spectrum beta-lactamase producing enterobacterial isolates in a Malaysian teaching hospital
    Ibrahim N, Wajidi MF, Yusof MY, Tay ST
    Trop Biomed, 2011 Dec;28(3):668-71.
    PMID: 22433898 MyJurnal
    The increased frequency of antibiotic resistance is known to be associated with the dissemination of integrons in the Enterobacteriaceae. This study determined the prevalence and type of integrons amongst 160 extended-spectrum beta-lactamase producing enterobacterial isolates kept in our culture collection. Integrons were detected in 98(61.3%) isolates, including 28(62.2%) Escherichia coli, 34(64.2%) Klebsiella spp., 27(61.4%), Enterobacter spp. and 9(50.0%) Citrobacter spp. investigated in this study. Restriction analysis of the integron gene fragments revealed that class I integron was the principal integron detected in 92(57.5%) of our isolates. Class II integron was detected in 6(3.8%) of our isolates, while no class III integron was detected in this study. The high rates of integron prevalence particularly of the class I integron in the E. coli and Klebsiella spp. concur with previous studies in other geographical regions. The higher (≥50%) integron prevalence of Citrobacter and Enterobacter isolates comparing to previous studies suggests the potential of these isolates as sources for dissemination of resistance determinants. The finding in this study serves as a basis for further study on the antibiotic resistance mechanisms of enterobacterial species in this teaching hospital.
  5. Fulltext Rickettsial infections in monkeys, Malaysia
    Tay ST, Koh FX, Kho KL, Sitam FT
    Emerg Infect Dis, 2015 Mar;21(3):545-7.
    PMID: 25695615 DOI: 10.3201/eid2103.141457
  6. Revisiting oral thrush in South-East Asian patients: A review of published studies (2000-2020)
    Karajacob AS, Al-Maleki AR, Tay ST
    J Oral Pathol Med, 2022 Jan;51(1):98-105.
    PMID: 34859509 DOI: 10.1111/jop.13266
    BACKGROUND: Oral thrush, a clinical condition due to an overgrowth of Candida yeasts in the oral cavity, is prominent in patients with immunosuppression. As recent updates on oral thrush in South-East Asian (SEA) countries are lacking, this review aimed to address the epidemiology, clinical features and distribution of Candida species, based on published studies in SEA countries over the last two decades.

    METHODS: Published studies on oral candidiasis (2000-2020) were retrieved from PubMed, Scopus, ISI Web of Science and Google Scholar databases to provide information on the incidence and factors affecting oral thrush cases in SEA countries.

    RESULTS: A total of 22 cross-sectional studies involving 3697 subjects from five SEA countries were reviewed in this study. The most frequently reported population were human immunodeficiency virus (HIV)-infected patients. The overall incidence rates amongst HIV-infected patients ranged from 20.7% to 97.0%, while incidence rates ranging from 0% to 72.7% were recorded for non-HIV-infected populations. Pseudomembranous candidiasis and erythematous candidiasis were the most common clinical presentations of oral thrush lesions. Candida albicans was the most common species identified in SEA studies. As oral thrush assessments were made merely based on clinical diagnosis, culture results were not available for most studies.

    CONCLUSION: This review highlights that most studies reporting on oral candidiasis in SEA countries were based on HIV-positive patients. Data are still lacking on oral candidiasis amongst non-HIV immunocompromised and immunocompetent patients. Increasing awareness on the diagnosis, treatment and consequences of this infection, and improved laboratory methods are essential for the management of oral candidiasis in this region.

  7. Prevention practices of vaginitis among Malaysian women and its associated factors
    Wan Muda WM, Wong LP, Tay ST
    J Obstet Gynaecol, 2018 Jul;38(5):708-715.
    PMID: 29526143 DOI: 10.1080/01443615.2017.1405923
    Information on vaginal-related issues among Malaysian women is very limited. This study aimed to explore factors associated with preventive practices of vaginitis among Malaysian women. A cross-sectional computer-assisted telephone interview survey of a representative sample of multi-racial Malaysian women aged 18-50 years old was conducted from January to April 2014. Women from 1446 households responded to the survey and nearly one-third (32.1%) reported to have experienced vaginitis. In multivariate analyses, respondents in the urban locality were more likely to practice vaginitis prevention (OR = 1.40, 95% CI = 1.06-1.84) compared with those in the rural areas. Respondents who perceived low susceptibility to vaginitis were less likely to practice vaginitis prevention (OR = 0.72, 95% CI = 0.57-0.91) compared with responders who highly perceive susceptibility. Respondents who had no formal education were less likely to practice vaginitis prevention (OR = 0.16, 95% CI = 0.05-0.48) compared with those with tertiary education. This study showed that comprehensive education and health programmes need to focus on women with a low educational level, living in rural areas and women with low perceived susceptibility to vaginitis. Impact statement What is already known on this subject? Little is known about vaginitis issues among women in Malaysia. This study provides information regarding vaginitis among Malaysian women by looking at the factors associated with prevention practices. What do the results of this study add? From our study, factors associated with prevention practices were found to be educational level, locality, and perceived susceptibility of vaginitis. Those who perform fewer vaginitis prevention practices were women with a low educational level and those who live in rural areas. From the Health Belief Model, women with a low perceived susceptibility of vaginitis were less likely to carry out vaginitis prevention practices. What are the implications of these findings for clinical practice and/or further research? The findings may provide additional insights for policy makers and healthcare providers to deliver effective approaches in order to improve prevention practices of vaginitis among women in multi-ethnic communities. This study has identified points of interest which need to be put in attention for women's health section which has been overlooked in Malaysia.
  8. Fulltext Candida auris: A Mini Review on Epidemiology in Healthcare Facilities in Asia
    Thatchanamoorthy N, Rukumani Devi V, Chandramathi S, Tay ST
    J Fungi (Basel), 2022 Oct 26;8(11).
    PMID: 36354893 DOI: 10.3390/jof8111126
    Candida auris, a newly emerging healthcare-associated yeast pathogen from the Metschnikowiaceae family, was first described in the ear canal of an elderly Japanese patient in 2009. The yeast is one of the causative agents of candidemia, which has been linked with nosocomial outbreaks and high mortality rates in healthcare facilities worldwide. Since its first isolation, the occurrence of C. auris in six continents has becomes a grave concern for the healthcare professionals and scientific community. Recent reports showed the identification of five geographically distinct clades and high rates of antifungal resistance associated with C. auris. Till date, there are no effective treatment options, and standardized measures for prevention and control of C. auris infection in healthcare facilities. This leads to frequent therapeutic failures and complicates the eradication of C. auris infection in healthcare facilities. Thus, this review focuses on the recent understanding of the epidemiology, risk factors, diagnosis, transmission and prevention and control strategies of C. auris infection in healthcare facilities in Asia.
  9. Fulltext Correction: First isolation and identification of Cystobasidium calyptogenae from the oral samples of an elderly patient presenting with angular cheilitis
    Karajacob AS, Goh JPE, Kallarakkal TG, Tay ST
    Eur J Med Res, 2022 Dec 06;27(1):277.
    PMID: 36471451 DOI: 10.1186/s40001-022-00884-9
  10. In vitro susceptibilities of Malaysian clinical isolates of Cryptococcus neoformans var. grubii and Cryptococcus gattii to five antifungal drugs
    Tay ST, Tanty Haryanty T, Ng KP, Rohani MY, Hamimah H
    Mycoses, 2006 Jul;49(4):324-30.
    PMID: 16784448
    The in vitro susceptibilities of Malaysian clinical isolates of Cryptococcus neoformans var. grubii and C . gattii to five antifungal drugs (amphotericin B, flucytosine, fluconazole, itraconazole and ketoconazole) were determined using the Etest method. None of the Malaysian isolates was resistant to amphotericin B and ketoconazole. Isolates resistant to flucytosine, fluconazole and itraconazole were observed in this study. Minimum inhibition concentrations (MICs) of > or = 32 microg ml(-1) against flucytosine, > or = 64 microg ml(-1) against fluconazole and > or = 1 microg ml(-1) against itraconazole were noted in four (8.3%), two (4.2%) and one (2.1%) isolates respectively. There was no significant difference in the MICs for both Cryptococcus species (P > 0.05), indicating that C. gattii was as susceptible as var. grubii to all the antifungal drugs tested. No significant difference in the MICs for both Cryptococcus species collected from 1980 to 1990 and 2002 to 2004 were observed (P > 0.05).
  11. Epidemiology of cryptococcosis in Malaysia
    Tay ST, Rohani MY, Hoo TS, Hamimah H
    Mycoses, 2010 Nov;53(6):509-14.
    PMID: 19627508 DOI: 10.1111/j.1439-0507.2009.01750.x
    This study describes the isolation of Cryptococcus neoformans and Cryptococcus gattii from patients with chronic meningitis who were admitted to 16 Malaysian hospitals, from 2003 to 2004. Of the 96 cryptococcal cases reported over the 2-year period, 74 (77.1%) patients were male and 45 (46.9%) patients were between 30 and 39 years old. Cryptococcosis was uncommon in children. A total of 57 (59.4%) and 23 (24.0%) patients were Malay and Chinese respectively. Human immunodeficiency virus infection was the major underlying disease reported in 36 (37.5%) patients. C. neoformans var. grubii (serotype A and molecular type VNI) was the predominant Cryptococcus species isolated from 88.5% of cryptococcal cases in this country. Cryptococcal cases due to C. neoformans var. grubii were reported from all the five regions in Malaysia, with the most number of cases reported from the central and northern regions. Cryptococcus gattii (all were serotype B and molecular types VGI/II) was isolated from all regions except the southern region. Compared with a study conducted prior to the AIDS era, our findings show substantial changes in the demographical characteristics of patients.
  12. Fulltext Short report: Predictors of severe disease in melioidosis patients in Kuala Lumpur, Malaysia
    Mohd Roslani AD, Tay ST, Puthucheary SD, Rukumani DV, Sam IC
    Am J Trop Med Hyg, 2014 Dec;91(6):1176-8.
    PMID: 25246695 DOI: 10.4269/ajtmh.14-0354
    The predictors of severe disease or death were determined for 85 melioidosis patients in Kuala Lumpur, Malaysia. Most of the patients were male, > 40 years old, and diabetic. Severe disease or death occurred in 28 (32.9%) cases. Lower lymphocyte counts and positive blood cultures were significant independent predictors of severe disease, but age, presentations with pneumonia, inappropriate empirical antibiotics, or flagellin types of the infecting isolates were not. Knowledge of local predictors of severe disease is useful for clinical management.
  13. Occurrence and characterization of Candida nivariensis from a culture collection of Candida glabrata clinical isolates in Malaysia
    Tay ST, Lotfalikhani A, Sabet NS, Ponnampalavanar S, Sulaiman S, Na SL, et al.
    Mycopathologia, 2014 Oct;178(3-4):307-14.
    PMID: 25022264 DOI: 10.1007/s11046-014-9778-9
    BACKGROUND: Candida nivariensis and C. bracarensis have been recently identified as emerging yeast pathogens which are phenotypically indistinguishable from C. glabrata. However, there is little data on the prevalence and antifungal susceptibilities of these species.

    OBJECTIVE: This study investigated the occurrence of C. nivariensis and C. bracarensis in a culture collection of 185 C. glabrata isolates at a Malaysian teaching hospital.

    METHODS: C. nivariensis was discriminated from C. glabrata using a PCR assay as described by Enache-Angoulvant et al. (J Clin Microbiol 49:3375-9, 2011). The identity of the isolates was confirmed by sequence analysis of the D1D2 domain and internal transcribed spacer region of the yeasts. The isolates were cultured on Chromogenic CHROMagar Candida (®) agar (Difco, USA), and their biochemical and enzymic profiles were determined. Antifungal susceptibilities of the isolates against amphotericin B, fluconazole, voriconazole and caspofungin were determined using E tests. Clotrimazole MICs were determined using a microbroth dilution method.

    RESULTS: There was a low prevalence (1.1 %) of C. nivariensis in our culture collection of C. glabrata. C. nivariensis was isolated from a blood culture and vaginal swab of two patients. C. nivariensis grew as white colonies on Chromogenic agar and demonstrated few positive reactions using biochemical tests. Enzymatic profiles of the C. nivariensis isolates were similar to that of C. glabrata. The isolates were susceptible to amphotericin B, fluconazole, voriconazole and caspofungin. Clotrimazole resistance is suspected in one isolate.

    CONCLUSION: This study reports for the first time the emergence of C. nivariensis in our clinical setting.

  14. Enhanced intracellular survival and epithelial cell adherence abilities of Burkholderia pseudomallei morphotypes are dependent on differential expression of virulence-associated proteins during mid-logarithmic growth phase
    Al-Maleki AR, Mariappan V, Vellasamy KM, Shankar EM, Tay ST, Vadivelu J
    J Proteomics, 2014 Jun 25;106:205-20.
    PMID: 24742602 DOI: 10.1016/j.jprot.2014.04.005
    Colony morphology variation is a characteristic of Burkholderia pseudomallei primary clinical isolates, associated with variations in expression of virulence factors. Here, we performed comparative investigations on adhesion, invasion, plaque-forming abilities and protein profiles of B. pseudomallei wild-type (WT) and a small colony variant (SCV). The percentage of SCV adherence to A549 cells was significantly higher (2.73%) than WT (1.91%). In contrast, WT was significantly more efficient (0.63%) than SCV (0.31%) in invasiveness and in inducing cellular damage. Using 2-DE and MALDI TOF/TOF, 263 and 258 protein spots were detected in WT and SCV, respectively. Comparatively, 49 proteins were differentially expressed in SCV when compared with WT. Of these, 31 proteins were up-regulated, namely, nucleoside diphosphate kinase (Ndk), phosphoglycerate kinase (Pgk), thioredoxin (TrxA), putative ferritin DPS-family DNA-binding protein (DPS) and oxidoreductase (AhpC) that are known to be involved in adhesion, intracellular survival and persistence. However, among the 18 down-regulated proteins, enolase (Eno), elongation factor (EF-Tu) and universal stress-related proteins were associated with invasion and virulence. Differences observed in these protein profiles provide ample clues to their association with the morphotypic and phenotypic characteristics of colony variants, providing additional insights into the potential association of B. pseudomallei colony morphotypes with disease pathogenesis.
  15. Fulltext Phenotypic detection of metallo-β-lactamase in imipenem-resistant Pseudomonas aeruginosa
    Khosravi Y, Loke MF, Chua EG, Tay ST, Vadivelu J
    ScientificWorldJournal, 2012;2012:654939.
    PMID: 22792048 DOI: 10.1100/2012/654939
    Carbapenems are the primary choice of treatment for severe Pseudomonas aeruginosa infection. However, the emergence of carbapenem resistance due to the production of metallo-β-lactamases (MBLs) is of global concern. In this study, 90 imipenem- (IPM- or IP-) resistant P. aeruginosa (IRPA) isolates, including 32 previously tested positive and genotyped for MBL genes by PCR, were subjected to double-disk synergy test (DDST), combined disk test (CDT), and imipenem/imipenem-inhibitor (IP/IPI) E-test to evaluate their MBLs detection capability. All three methods were shown to have a sensitivity of 100%. However, DDST was the most specific of the three (96.6%), followed by IP/IPI E-test interpreted based on the single criteria of IP/IPI ≥8 as positive (62.1%), and CDT was the least specific (43.1%). Based on the data from this evaluation, we propose that only IRPA with IP MIC >16 μg/mL and IP/IPI ≥8 by IP/IPI E-test should be taken as positive for MBL activity. With the new dual interpretation criteria, the MBL IP/IPI E-test was shown to achieve 100% sensitivity as well as specificity for the IRPA in this study. Therefore, the IP/IPI E-test is a viable alternative phenotypic assay to detect MBL production in IRPA in our population in circumstances where PCR detection is not a feasible option.
  16. Development of a multiplex PCR assay for rapid identification of Burkholderia pseudomallei, Burkholderia thailandensis, Burkholderia mallei and Burkholderia cepacia complex
    Koh SF, Tay ST, Sermswan R, Wongratanacheewin S, Chua KH, Puthucheary SD
    J Microbiol Methods, 2012 Sep;90(3):305-8.
    PMID: 22705921 DOI: 10.1016/j.mimet.2012.06.002
    We have developed a multiplex PCR assay for rapid identification and differentiation of cultures for Burkholderia pseudomallei, Burkholderia thailandensis, Burkholderia mallei and Burkholderia cepacia complex. The assay is valuable for use in clinical and veterinary laboratories, and in a deployable laboratory during outbreaks.
  17. The isolation, characterization and antifungal susceptibilities of Cryptococcus neoformans from bird excreta in Klang Valley, Malaysia
    Tay ST, Chai HC, Na SL, Hamimah H, Rohani MY, Soo-Hoo TS
    Mycopathologia, 2005 Jun;159(4):509-13.
    PMID: 15983736
    The occurrence of Cryptococcus neoformans in bird excreta in Klang valley, Malaysia was determined in this study. Of 544 samples of bird excreta collected from a local zoo, pet shops and public areas, 20 strains of C. neoformans were isolated. All C. neoformans strains were serotype A and thus identified as C. neoformans variety grubii. All did not produce color changes on canavanine-glycine-bromothymol blue agar. All were of alpha-mating types, as determined by a pheromone-specific PCR assay. The antifungal susceptibility testing using agar diffusion method Neo-sensitabs showed that all were susceptible to amphotericin B, fluconazole and itraconazole.
  18. Fulltext Genome-Wide Transcription Study of Cryptococcus neoformans H99 Clinical Strain versus Environmental Strains
    Movahed E, Munusamy K, Tan GM, Looi CY, Tay ST, Wong WF
    PLoS One, 2015;10(9):e0137457.
    PMID: 26360021 DOI: 10.1371/journal.pone.0137457
    The infection of Cryptococcus neoformans is acquired through the inhalation of desiccated yeast cells and basidiospores originated from the environment, particularly from bird's droppings and decaying wood. Three environmental strains of C. neoformans originated from bird droppings (H4, S48B and S68B) and C. neoformans reference clinical strain (H99) were used for intranasal infection in C57BL/6 mice. We showed that the H99 strain demonstrated higher virulence compared to H4, S48B and S68B strains. To examine if gene expression contributed to the different degree of virulence among these strains, a genome-wide microarray study was performed to inspect the transcriptomic profiles of all four strains. Our results revealed that out of 7,419 genes (22,257 probes) examined, 65 genes were significantly up-or down-regulated in H99 versus H4, S48B and S68B strains. The up-regulated genes in H99 strain include Hydroxymethylglutaryl-CoA synthase (MVA1), Mitochondrial matrix factor 1 (MMF1), Bud-site-selection protein 8 (BUD8), High affinity glucose transporter 3 (SNF3) and Rho GTPase-activating protein 2 (RGA2). Pathway annotation using DAVID bioinformatics resource showed that metal ion binding and sugar transmembrane transporter activity pathways were highly expressed in the H99 strain. We suggest that the genes and pathways identified may possibly play crucial roles in the fungal pathogenesis.
  19. Fulltext Triclosan Demonstrates Synergic Effect with Amphotericin B and Fluconazole and Induces Apoptosis-Like Cell Death in Cryptococcus neoformans
    Movahed E, Tan GM, Munusamy K, Yeow TC, Tay ST, Wong WF, et al.
    Front Microbiol, 2016;7:360.
    PMID: 27047474 DOI: 10.3389/fmicb.2016.00360
    Cryptococcus neoformans is an opportunistic fungus that causes fatal meningoencephalitis especially in AIDS patients. There is an increasing need for discovery of new anti-cryptococcal drugs due to emergence of resistance cases in recent years. In this study, we aim to elucidate the antifungal effect of triclosan against C. neoformans.
  20. Fulltext Corrigendum to "Phenotypic Detection of Metallo-β-Lactamase in Imipenem-Resistant Pseudomonas aeruginosa"
    Khosravi Y, Loke MF, Chua EG, Tay ST, Vadivelu J
    ScientificWorldJournal, 2016;2016:9562039.
    PMID: 27314061
    [This corrects the article DOI: 10.1100/2012/654939.].
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