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Synthesis and Staphylococcus aureus biofilm inhibitory activity of indolenine-substituted pyrazole and pyrimido[1,2-b]indazole derivatives

Yap CH, Ramle AQ, Lim SK, Rames A, Tay ST, Chin SP, et al.

Bioorg Med Chem, 2023 Nov 15;95:117485.
PMID: 37812886 DOI: 10.1016/j.bmc.2023.117485

Staphylococcus aureus is a highly adaptable opportunistic pathogen that can form biofilms and generate persister cells, leading to life-threatening infections that are difficult to treat with antibiotics alone. Therefore, there is a need for an effective S. aureus biofilm inhibitor to combat this public health threat. In this study, a small library of indolenine-substituted pyrazoles and pyrimido[1,2-b]indazole derivatives were synthesised, of which the hit compound exhibited promising antibiofilm activities against methicillin-susceptible S. aureus (MSSA ATCC 29213) and methicillin-resistant S. aureus (MRSA ATCC 33591) at concentrations significantly lower than the planktonic growth inhibition. The hit compound could prevent biofilm formation and eradicate mature biofilms of MSSA and MRSA, with a minimum biofilm inhibitory concentration (MBIC50) value as low as 1.56 µg/mL and a minimum biofilm eradication concentration (MBEC50) value as low as 6.25 µg/mL. The minimum inhibitory concentration (MIC) values of the hit compound against MSSA and MRSA were 50 µg/mL and 25 µg/mL, respectively, while the minimum bactericidal concentration (MBC) values against MSSA and MRSA were > 100 µg/mL. Preliminary structure-activity relationship analysis reveals that the fused benzene ring and COOH group of the hit compound are crucial for the antibiofilm activity. Additionally, the compound was not cytotoxic to human alveolar A549 cells, thus highlighting its potential as a suitable candidate for further development as a S. aureus biofilm inhibitor.
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.
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.
Whole-genome sequence analysis and exploration of the zoonotic potential of a rat-borne Bartonella elizabethae

Tay ST, Kho KL, Wee WY, Choo SW

Acta Trop, 2016 Mar;155:25-33.
PMID: 26658020 DOI: 10.1016/j.actatropica.2015.11.019

Bartonella elizabethae has been known to cause endocarditis and neuroretinitis in humans. The genomic features and virulence profiles of a B. elizabethae strain (designated as BeUM) isolated from the spleen of a wild rat in Kuala Lumpur, Malaysia are described in this study. The BeUM strain has a genome size of 1,932,479bp and GC content of 38.3%. There is a high degree of conservation between the genomes of strain BeUM with B. elizabethae type strains (ATCC 49927 and F9251) and a rat-borne strain, Re6043vi. Of 2137 gene clusters identified from B. elizabethae strains, 2064 (96.6%) are indicated as the core gene clusters. Comparative genome analysis of B. elizabethae strains reveals virulence genes which are known in other pathogenic Bartonella species, including VirB2-11, vbhB2-B11, VirD4, trw, vapA2-5, hbpA-E, bepA-F, bepH, badA/vomp/brp, ialB, omp43/89 and korA-B. A putative intact prophage has been identified in the strain BeUM, in addition to a 8kb pathogenicity island. The whole genome analysis supports the zoonotic potential of the rodent-borne B. elizabethae, and provides basis for future functional and pathogenicity studies of B. elizabethae.
Fulltext Growth inhibition of Candida species by Wickerhamomyces anomalus mycocin and a lactone compound of Aureobasidium pullulans

Tay ST, Lim SL, Tan HW

BMC Complement Altern Med, 2014;14:439.
PMID: 25380692 DOI: 10.1186/1472-6882-14-439

The increasing resistance of Candida yeasts towards antifungal compounds and the limited choice of therapeutic drugs have spurred great interest amongst the scientific community to search for alternative anti-Candida compounds. Mycocins and fungal metabolites have been reported to have the potential for treatment of fungal infections. In this study, the growth inhibition of Candida species by a mycocin produced by Wickerhamomyces anomalus and a lactone compound from Aureobasidium pullulans were investigated.
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.
Fulltext Isolation and molecular identification of Bartonellae from wild rats (Rattus species) in Malaysia

Tay ST, Mokhtar AS, Zain SN, Low KC

Am J Trop Med Hyg, 2014 Jun;90(6):1039-42.
PMID: 24732465 DOI: 10.4269/ajtmh.13-0273

This study describes our investigation on the prevalence and molecular identification of bartonellae from Rattus diardii and R. norvegicus in the urban areas of Malaysia. Of 95 rats investigated, Bartonella tribocorum, B. rattimassiliensis, B. coopersplainsensis, B. elizabethae, and B. queenslandensis were isolated from kidney and spleen homogenates of four rats. Bartonellae DNA was amplified from the rat organ tissues by using primers specific for the bartonellae RNA polymerase beta subunit (rpoB) gene in nine other rats. Sequence analysis of the rpoB gene fragments shows the identification of B. queenslandensis in five rats, B. elizabethae in three rats, and B. tribocorum in one rat. Combining the results of isolation and molecular detection of bartonellae, we found that the prevalence of Bartonella infection in the Rattus spp. investigated in this study was 13.7%. Implementation of effective rat control program in the urban areas is necessary to prevent the spillover of bartonellosis from rats to humans.
Fulltext Antibody prevalence and factors associated with exposure to Orientia tsutsugamushi in different aboriginal subgroups in West Malaysia

Tay ST, Mohamed Zan HA, Lim YA, Ngui R

PLoS Negl Trop Dis, 2013;7(8):e2341.
PMID: 23936576 DOI: 10.1371/journal.pntd.0002341

Limited data is available on the current status of scrub typhus infection in the aboriginal population in Malaysia. This study was aimed to provide recent data on the degree of exposure of 280 individuals from seven aboriginal subgroups to Orientia tsutsugamushi (causative agent of scrub typhus) in West Malaysia. The environment, socioeconomic and behavioural risk factors associated with the disease were also investigated.
Sequence analysis of the hypervariable regions of the 56 kDa immunodominant protein genes of Orientia tsutsugamushi strains in Malaysia

Tay ST, Rohani YM, Ho TM, Shamala D

Microbiol. Immunol., 2005;49(1):67-71.
PMID: 15665455

The DNA sequences encompassing two hypervariable regions, VD II and III of the 56 kDa immunodominant protein gene of 21 Malaysian strains of Orientia tsutsugamushi were determined. Two strains demonstrated a 100% DNA homology with the Gilliam prototype strain, and one with TH1817 strain and TA678 strain respectively. High percentages of DNA similarity (95-99%) were observed with Karp (4 strains), Gilliam (2 strains), TH1817 (4 strains), TC586 (3 strains) and TA763 (1 strain). The remaining strains demonstrated the highest DNA similarity with TA763 (1 strain, 89%), TA678 (1 strain, 86%) and TA686 (1 strain, 87%). Our study provides additional evidence on the existence and the genetic heterogeneity of TA strains of the Southeast Asia and their closely related strains in Malaysia.
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.
Molecular subtyping of clinical isolates of Candida albicans and identification of Candida dubliniensis Malaysia

Tay ST, Chai HC, Na SL, Ng KP

Mycopathologia, 2005 Apr;159(3):325-9.
PMID: 15883714

The genotypes of 221 recent isolates of Candida albicans from various clinical specimens of 213 patients admitted to the University Malaya Medical Centre, Malaysia was determined based on the amplification of a transposable intron region in the 25 S rRNA gene. The analyses of 178 C. albicans isolated from nonsterile clinical specimens showed that they could be classified into three genotypes: genotype A (138 isolates), genotype B (38 isolates) and genotype C (2 isolates). The genotyping of 43 clinical isolates from sterile specimens showed that they belonged to genotype A (29 isolates), genotype B (10 isolates), genotype C (2 isolates) and genotype D (2 isolates). The overall distribution of C. albicans genotypes in sterile and nonsterile specimens appeared similar, with genotype A being the most predominant type. This study reported the identification of C. dubliniensis (genotype D) in 2 HIV-negative patients with systemic candidiasis, which were missed by the routine mycological procedure. The study demonstrated the genetic diversity of clinical isolates of C. albicans in Malaysia.
Isolation and PCR detection of rickettsiae from clinical and rodent samples in Malaysia

Tay ST, Rohani MY, Ho TM, Devi S

Southeast Asian J. Trop. Med. Public Health, 2002 Dec;33(4):772-9.
PMID: 12757225

Isolation of rickettsiae from patients' blood samples and organ samples of wild rodents from areas with high seroprevalence of rickettsial infections was attempted using cell culture assay and animal passages. L929 mouse fibroblast cells grown in 24 well tissue culture plate were inoculated with buffy coat of febrile patients and examined for the growth of rickettsiae by Giemsa, Gimenez staining and direct immunofluorescence assay. No rickettsiae were isolated from 48 patients' blood samples. No symptomatic infections were noted in mice or guinea pigs infected with 50 organ samples of wild rodents. There was no rickettsial DNA amplified from these samples using various PCR detection systems for Orientia tsutsugamushi, typhus and spotted fever group rickettsiae.
Borrelia burgdorferi (strain B. afzelii) antibodies among Malaysian blood donors and patients

Tay ST, Kamalanathan M, Rohani MY

Southeast Asian J. Trop. Med. Public Health, 2002 Dec;33(4):787-93.
PMID: 12757227

In this study, the presence of IgG and IgM antibodies against Borrelia burgdorferi (strain B. afzelii) among Malaysian blood donors and patients admitted to hospital with various infectious diseases was determined. Sera were screened using enzyme-linked immunosorbent assay (ELISA); positive sera were then subjected to Western blot testing. All but one of the blood donors were negative for borrelial antibodies. Of 121 patients' sera, IgM antibodies were detected in 24 (19.8%) and IgG antibodies were detected in 5 (4.1%) sera. Only one of two patients with skin manifestations suggestive of Lyme disease had IgM antibody against B. afzelii. Of 30 patients with exposure to tick typhus, 4 (13.3%) were IgM positive and 1 (3.3%) was IgG positive. Based on the detection of antigenic bands by Western blot, 6 patients' sera showed positive reactions. Antigenic bands of p39, p41 and p59/62 kDa were the commonest findings of Western blotting. This study provides serological evidence of B. afzelii infections in Malaysia; further investigation is needed to correlate serological and clinical findings.
Virulence of Malaysian isolates of Orientia tsutsugamushi in mice

Tay ST, Rohani MY, Ho TM, Devi S

Southeast Asian J. Trop. Med. Public Health, 2002 Sep;33(3):565-9.
PMID: 12693592

The pathogenicity of Malaysian isolates of Orientia tsutsugamushi was investigated by a mouse virulence assay. The isolates could be differentiated as low (4 isolates), moderately (3 isolates) and highly virulent (2 isolates) based on the different responses in infected mice. No direct correlation between severity of human scrub typhus infections and virulence of the O. tsutsugamushi in mice was observed. Mice infected with virulent strains of O. tsutsugamushi showed splenomegaly, ascitis accumulation and enlargement of kidneys and livers whereas avirulent O. tsutsugamushi strains were asymptomatic and exhibited ruffled fur for a short period after infection. There was low antibody response in mice infected with isolates of low pathogenicity as compared with those of highly virulent isolates. Upon dissection of the infected mice, enlargement of mouse organs such as spleen, kidney and liver was noted. Presence of rickettsemia in mice was confirmed by the growth of O. tsutsugamushi in the L929 cells when inoculated with blood from infected mice. O. tsutsugamushi was also cultured from the peritoneal exudates of the infected mice. However, DNA of O. tsutsugamushi was only detected in the peritoneal exudates (by PCR) and blood (by cell culture) and not from other tissue samples.
Antigenic types of Orientia tsutsugamushi in Malaysia

Tay ST, Rohani MY, Ho TM, Devi S

Southeast Asian J. Trop. Med. Public Health, 2002 Sep;33(3):557-64.
PMID: 12693591

The seroprevalence of various Orientia tsutsugamushi (OT) strains among Malaysian patients with suspected scrub typhus infections was determined using an indirect immunoperoxidase (IIP) assay. IgG against a single OT strain were detected in six sera (3 Karp, 1 Gilliam and 2 TC586), whereas IgM antibodies against a single OT strain (Gilliam) were noted in 3 sera (Gilliam). IgG reactive to all OT strains were present in 33 (47.1%) of the 70 sera and IgM reactive to all OT strains were present in 22 (78.6%) of the 28 sera. The fact that most sera were reactive to multiple OT strains suggests that group-specific antigens are involved in scrub typhus infections, whereas very few were due to strain-specific epitopes present on these strains. Peak IgG and IgM titers were noted more frequently against Gilliam, Karp, and TA763 strains: this suggests that these strains may be the commonest infecting strains among Malaysian patients. Two predominant OT polypeptides consistently reacted with patients' sera were the 70 kDa and 56 kDa proteins.
The use of the indirect immunoperoxidase test for the serodiagnosis of rickettsial diseases in Malaysia

Tay ST, Rohani MY

Southeast Asian J. Trop. Med. Public Health, 2002 Jun;33(2):314-20.
PMID: 12236431

The indirect immunoperoxidase (HP) test has been used extensively in most government hospitals in Malaysia for the serodiagnosis of scrub typhus, murine typhus and tick typhus during the 1990s. The test was used to determine the IgG and IgM antibody titers in patients' sera for three rickettsial species, ie Orientia tsutsugamushi OT; the causative agent of scrub typhus), Rickettsia typhi (RT; the causative agent of murine typhus), and TT118 spotted fever group rickettsiae (TT; the causative agent of tick typhus). The serological findings obtained from Malaysian hospitals using the IIP test (1994-1999) were analyzed. During the six-year period, a total of 61,501 patients' sera were tested, of which 9.6%, 10.5%, and 12.9% had antibody (IgG and/or IgM of > or = 1:50) for OT, RT and TT respectively. A total of 8.6%, 9.8%, and 9.7% of sera had IgG antibody of > or = 1:50 for OT, RT, and TT respectively, indicating past infection. A total of 3.4%, 3.8%, and 6.4 % of sera had IgM antibody of > or = 1:50 for OT, RT, and TT respectively, indicating recent infection. A total of 2,986 (4.9%), 1,882 (3.1%), and 1,574 (2.6%) of sera had IgG and/or IgM antibody titers of > or = 1:400 for OT, RT, and TT respectively, suggesting active rickettsial infection. The seropositivity rates of OT, RT and TT varied according to geographical locations. While the seropositivity of OT remained constant during the six-year period, a reduction in the seropositivity of both RT and TT was noted during recent years. The serological findings reflect the endemicity of rickettsial diseases, including tick typhus, and endemic typhus in various parts of Malaysia. Awareness of these diseases by health and medical staff and by the general public is important if the mortality and morbidity associated with scrub typhus, tick typhus, and murine typhus in Malaysia, are to be reduced.
Fulltext Prevalence and antibiotic susceptibility of Ureaplasma urealyticum in Malaysian neonates with respiratory distress

Tay ST, Boo NY, Khoo TB, Koay AS, Rohani MY

Med J Malaysia, 1997 Dec;52(4):409-11.
PMID: 10968119

Ureaplasma urealyticum was isolated from the endotracheal aspirates of 39 (21.4%) of 182 neonates with respiratory distress requiring ventilatory support. Mycoplasma hominis was isolated from one (0.5%) neonate. Bacterial cultures were negative in 123 (67.6%) neonates. Antibiotic susceptibility test carried out on ten isolates of U. urealyticum showed that all the organisms were sensitive to erythromycin but resistant to lincomycin and sulfamethoxazole trimethoprim. All, except one, U. urealyticum were sensitive to tetracycline and minocycline. Two isolates were resistant to ciprofloxacin. This study showed that U.urealyticum was a common organism isolated from the endotracheal aspirates of neonates with respiratory distress.
Expression of recombinant proteins of Orientia tsutsugamushi and their applications in the serodiagnosis of scrub typhus

Tay ST, Rohani MY, Ho TM, Devi S

Diagn Microbiol Infect Dis, 2002 Oct;44(2):137-42.
PMID: 12458119

In this study, recombinant proteins that encompassed the AD I-AD III regions of 56 kDa immunodominant gene of 2 Orientia tsutsugamushi (OT) serotypes; Gilliam and TA763 were expressed in Escherichia coli. Both recombinant proteins exhibited serologic cross-reactivity with the rabbit antisera against various OT serotypes, as evaluated by enzyme-linked immunosorbent assay (ELISA), but not against other rickettsial species, including Rickettsia typhi, R. prowazekii and TT118 SFG rickettsiae. The feasibility of using the recombinant proteins as a diagnostic reagent was further evaluated by ELISA using sera from blood donors and scrub typhus patients. The results suggested a higher affinity of the antihuman IgM than IgG with both recombinant proteins. The IgM ELISA findings were agreeable with the results of indirect immunoperoxidase (IIP) assay especially with sera of high antibody (1:1600). However, more than one antigen are probably needed for development of an effective assay for serodiagnosis of scrub typhus in endemic areas.
Isolation and polymerase chain reaction detection of Mycoplasma pneumoniae in Malaysian patients with respiratory tract infections

Tay ST, Habsah MY, Tan SC, Rohani MY

Southeast Asian J. Trop. Med. Public Health, 2000 Dec;31(4):688-92.
PMID: 11414413

Isolation and polymerase chain reaction (PCR) were performed for detection of Mycoplasma pneumoniae from respiratory tract specimens obtained from 200 adult and 200 pediatric patients. M. pneumoniae was isolated from bronchoalveolar lavage fluid of 1(0.5%) adult patient and 4(2.0%) tracheal aspirates of pediatric patients. PCR was positive for only one (0.5%) broncoalveolar lavage fluid of an adult patient and fifteen (7.5%) tracheal aspirates of pediatric patients. This study suggested that M. pneumoniae was more frequently detected in pediatric patients and PCR appears to have advantages over isolation, in terms of rapidity and sensitivity.
Detection of Mycoplasma pneumoniae from cerebrospinal fluid samples of pediatric patients

Tay ST, Habsah MY, Tan SC, Rohani MY

Southeast Asian J. Trop. Med. Public Health, 2000 Dec;31(4):684-7.
PMID: 11414412

Central nervous system manifestations are probably the most frequent extrapulmonary complications of infections due to Mycoplasma pneumoniae, occur mostly in children. In this study, we attempted to isolate M. pneumoniae and to detect the organism by polymerase chain reaction (PCR) from cerebrospinal fluid samples (CSF) of pediatric patients. Of the 244 CSF samples, no M. pneumoniae was isolated. Six (2.5%) of the CSF samples were positive by PCR amplification. More effort are necessary to isolate the organism from CSF samples in order to ascertain the role of M. pneumoniae in causing neurological complications.

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