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INTRODUCTION: Maternal medicine is important and complex. It focuses on the care of mothers with comorbidities or medical disorders during pregnancy. To date, there are limited tools to assess clinicians' knowledge, attitude and practice (KAP) in this field. MATERIALS AND METHODS: This study aims to develop and validate a questionnaire measuring the KAP of clinicians towards maternal medicine. Literature reviews, item generation and multiple experts' reviews were conducted during the questionnaire development phase. Convenient sampling was undertaken for this cross-sectional survey. A total of 168 clinicians from one tertiary hospital and three health clinics participated in thepilot testing using the modified questionnaire. In addition, test-retest was performed on 30 participants to examine its reliability, whilst exploratory factor analysis (EFA) was undertaken to determine its construct validity. RESULTS: This finalised questionnaire contained 36 items with excellent content validity. Pertaining to test-retest reliability, all knowledge domain items showed Kappa values > 0.20, except item K29; attitude domain items overall Cronbach's alpha was 0.787 with corrected item-total correlation > 0.300 and lastly, all items in practice domain achieved intra-class correlation index > 0.700, except P2.5. EFA supported four factor structures, but six items were removed due to the following reasons: cross loading, negative or poor loading factor (< 0.3). CONCLUSION: Overall, this instrument has an acceptable psychometric property, content validity, internal reliability and construct validity. It is hope that this questionnaire would be validated in other populations and be used in future research to enrich our understanding of clinicians' KAP towards maternal medicine.

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[This corrects the article DOI: 10.3389/frobt.2023.1123153.].

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INTRODUCTION: Dengue fever is an arthropod-borne disease and has a wide clinical spectrum. It is hypothesised that dengue serotypes could be a possible factor for such phenomena and therefore be a possible predictor for the development of severe dengue. METHOD: A retrospective cohort study was done to explore the association between dengue serotypes and the various complications. All patients who underwent dengue serotyping from 1st January to 31st December 2018 in Tengku Ampuan Rahimah Hospital were selected. Serotypes were randomly done for admitted dengue patients. Notes were then retrieved for data collection. Secondary outcomes like length of stay and highest lactate level were also studied. Data analysis was done using SPSS version 20. RESULT: A total of 193 patient records were included in the analysis. Chi-square test for independence indicated that the proportion of dengue complications between male and female were significantly different (χ2(1) = 11.37, p = 0.001). Dengue serotype was not associated with the development of dengue complications, total number of dengue complications, length of admission, lactate level and survival among the serotypes. Results of the binary logistic regression showed that men have thrice the odds (AOR = 3.3, 95% CI: 1.6 6.7) for developing dengue complications. One patient was found to be co-infected with serotype 2 and 3. CONCLUSION: Our study did not reveal any association between the different dengue virus serotypes and its complications. Therefore, all dengue infection should be approached with equal meticulousness. There are possibilities that apart from serotype, dengue genotype and lineage would determine clinical outcome. However, more studies are required to study such associations.

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A Healthcare-assistive Infection-control RObot (HIRO) is a healthcare-assistive robot that is deployed in an outpatient primary care clinic to sanitise the premises, monitor people in its proximity for their temperature and donning of masks, and usher them to service points. This study aimed to determine the acceptability, perceptions of safety, and concerns among the patients, visitors, and polyclinic healthcare workers (HCWs) regarding the HIRO. A cross-sectional questionnaire survey was conducted from March to April 2022 when the HIRO was at Tampines Polyclinic in eastern Singapore. A total of 170 multidisciplinary HCWs serve approximately 1,000 patients and visitors daily at this polyclinic. The sample size of 385 was computed using a proportion of 0.5, 5% precision, and 95% confidence interval. Research assistants administered an e-survey to gather demographic data and feedback from 300 patients/visitors and 85 HCWs on their perceptions of the HIRO using Likert scales. The participants watched a video on the HIRO's functionalities and were given the opportunity to directly interact with it. Descriptive statistics was performed and figures were presented in frequencies and percentages. The majority of the participants viewed the HIRO's functionalities favourably: sanitising (96.7%/91.2%); checking proper mask donning (97%/89.4%); temperature monitoring (97%/91.7%); ushering (91.7%/81.1%); perceived user friendliness (93%/88.3%), and improvement in the clinic experience (96%/94.2%). A minority of the participants perceived harm from the HIRO's liquid disinfectant (29.6%/31.5%) and that its voice-annotated instructions may be upsetting (14%/24.8%). Most of the participants accepted the HIRO's deployment at the polyclinic and perceived it to be safe. The HIRO used ultraviolet irradiation for sanitisation during after-clinic hours instead of disinfectants due to the perceived harm.

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To validate survival of Lactobacillus casei strain Shirota (LcS) during passage through the gastrointestinal tract of healthy Singaporean young adults, 21 participants (18-25 years old) were asked to consume a 100 ml of fermented milk drink containing 1.0×108 cfu/ml of LcS daily for 14 days, and to maintain their dietary habit and life style. During and at the end of the ingestion period, both culture method (identity confirmed by ELISA) and 16s rRNA sequencing results revealed that viable LcS (7.27 and 7.64 log10 cfu/g of faeces at the ingestion period Day 7 and Day 14, respectively) and Lactobacillus could be recovered from the faeces of all the subjects. The viable LcS count from male and female were comparable for each time point. Before consumption (baseline) and 14 days after cessation of consumption of the fermented milk, LcS was not detected in most of the subjects. In this study condition, the composition of the major gut microbiota (>0.1% in relative abundance of genus) and characteristics of defaecation such as stool consistency and frequency of defecation did not change throughout the study before and after ingestion of LcS. LcS was able to survive passage through the gastrointestinal tract of Singapore adults without sustainable colonisation, but the effect of LcS on microbiota modulation, stool consistency and frequency was not observed under this study condition.

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Genetic diversity in human leukocyte antigen (HLA) molecules is thought to have arisen from the co-evolution between host and pathogen and maintained by balancing selection. Heterozygote advantage is a common proposed scenario for maintaining high levels of diversity in HLA genes, and extending from this, the divergent allele advantage (DAA) model suggests that individuals with more divergent HLA alleles bind and recognize a wider array of antigens. While the DAA model seems biologically suitable for driving HLA diversity, there is likely an upper threshold to the amount of sequence divergence. We used peptide-binding and pathogen-recognition capacity of DRB1 alleles as a model to further explore the DAA model; within the DRB1 locus, we examined binding predictions based on two distinct phylogenetic groups (denoted group A and B) previously identified based on non-peptide-binding region (PBR) nucleotide sequences. Predictions in this study support that group A allele and group B allele lineages have contrasting binding/recognition capacity, with only the latter supporting the DAA model. Furthermore, computer simulations revealed an inconsistency in the DAA model alone with observed extent of polymorphisms, supporting that the DAA model could only work effectively in combination with other mechanisms. Overall, we support that the mechanisms driving HLA diversity are non-exclusive. By investigating the relationships among HLA alleles, and pathogens recognized, we can provide further insights into the mechanisms on how humans have adapted to infectious diseases over time.

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Major histocompatibility complex class II (MHCII) genes code for proteins that bind and present antigenic peptides and trigger the adaptive immune response. We present a broad geographical study of MHCII DA ß1 (DAB) and DB ß1 (DBB) variants of the koala (Phascolarctos cinereus; n=191) from 12 populations across eastern Australia, with a total of 13 DAB and 7 DBB variants found. We identified greater MHCII variation and, possibly, additional gene copies in koala populations in the north (Queensland and New South Wales) relative to the south (Victoria), confirmed by STRUCTURE analyses and genetic differentiation using analysis of molecular variance. The higher MHCII diversity in the north relative to south could potentially be attributed to (i) significant founder effect in Victorian populations linked to historical translocation of bottlenecked koala populations and (ii) increased pathogen-driven balancing selection and/or local genetic drift in the north. Low MHCII genetic diversity in koalas from the south could reduce their potential response to disease, although the three DAB variants found in the south had substantial sequence divergence between variants. This study assessing MHCII diversity in the koala with historical translocations in some populations contributes to understanding the effects of population translocations on functional genetic diversity.

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APPL1 is an adaptor protein that binds to both AKT and adiponectin receptors and is hypothesised to mediate the effects of adiponectin in activating downstream effectors such as AMP-activated protein kinase (AMPK). We aimed to establish whether APPL1 plays a physiological role in mediating glycogen accumulation and insulin sensitivity in muscle and the signalling pathways involved. In vivo electrotransfer of cDNA- and shRNA-expressing constructs was used to over-express or silence APPL1 for 1 week in single tibialis cranialis muscles of rats. Resulting changes in glucose and lipid metabolism and signalling pathway activation were investigated under basal conditions and in high-fat diet (HFD)- or chow-fed rats under hyperinsulinaemic-euglycaemic clamp conditions. APPL1 over-expression (OE) caused an increase in glycogen storage and insulin-stimulated glycogen synthesis in muscle, accompanied by a modest increase in glucose uptake. Glycogen synthesis during the clamp was reduced by HFD but normalised by APPL1 OE. These effects are likely explained by APPL1 OE-induced increase in basal and insulin-stimulated phosphorylation of IRS1, AKT, GSK3ß and TBC1D4. On the contrary, APPL1 OE, such as HFD, reduced AMPK and acetyl-CoA carboxylase phosphorylation and PPARγ coactivator-1α and uncoupling protein 3 expression. Furthermore, APPL1 silencing caused complementary changes in glycogen storage and phosphorylation of AMPK and PI3-kinase pathway intermediates. Thus, APPL1 may provide a means for crosstalk between adiponectin and insulin signalling pathways, mediating the insulin-sensitising effects of adiponectin on muscle glucose disposal. These effects do not appear to require AMPK. Activation of signalling mediated via APPL1 may be beneficial in overcoming muscle insulin resistance.

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Medulloblastoma is the commonest primary central nervous system malignancy in childhood, but is rare in adults. Rarer still is metastasis of such a tumour to a lymph node. This report describes such a case, and briefly discusses relevant pathophysiology and epidemiology.

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Dermatofibrosarcoma protuberans (DFSP) is a rare slow-growing soft tissue tumour which characteristically occurs on the chest, head and neck regions. Typical histologic features include monomorphous spindle-shaped cells arranged in a storiform pattern on a background of fibrous stroma. The tumour has a propensity for local invasion and high recurrence rate. While the imaging features are not pathognomonic of the tumour, the characteristic location and shape should prompt inclusion in the differential diagnostic list.

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The objective was to evaluate the association of an abnormal second trimester prenatal biochemical screening with the subsecuent development of pregnancy complications in women carrying chromosomally normal fetuses. A prospective study of 123 pregnant patients was performed. Specimens were assayed for alpha-fetoprotein, unconjugated estriol, free alpha hCG, and total hCG. The study included the evaluation of the mean and standard desviation as well as multiple of the median. Six women (4.6%) had positive results. The frequency of pregnancy complications in this group was 33.3%, while in the group with negative screening was 11.1%. As conclusion, positive four marker screening is associated with a adverse pregnancy evolution. However the usefulness of four marker screening for to predict pregnancy complications needs more investigations.

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Signal transduction pathways involving the c-Raf protein kinase are frequently activated in tumor cells. We have addressed the relevance of this activation by a loss-of-function approach. An anti-sense phosphorothioate oligonucleotide (ODN) specifically targeted against c-raf mRNA (Monia et al., 1996a) was used to block c-Raf protein expression in four different cell lines derived from lung, cervical, prostate and colon carcinomas. Concomitant with the abrogation of c-Raf expression we observed the occurrence of classical apoptotic markers, including chromatin condensation, inter-nucleosomal DNA cleavage, annexin V binding and cleavage of PARP, which was followed by cell death, affecting most of the cell population. This induction of apoptosis occurred independent of the p53 status of the cell. These findings demonstrate that c-Raf can protect tumor cells from undergoing programmed cell death, and suggest that the interference with c-Raf expression or function by ODNs or specific drugs could represent a powerful means for improving the efficacy of anti-cancer therapy.

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OBJECTIVES: To employ polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis for the rapid differentiation of Neisseria gonorrhoeae protein IB (PIB) isolates and to compare its usefulness with the widely accepted auxotype/serovar classification scheme. METHODS: The outer membrane protein IB genes of 47 gonococcal isolates belonging to 10 different serovars were amplified by PCR. The approximately 1 kb DNA products were then digested separately with restriction enzymes CfoI and MspA1I, and electrophoresed on agarose gels. RESULTS: Cleavage of PIB genes by MspA1I and CfoI differentiated all the N gonorrhoeae strains into five and six PCR-RFLP profiles, respectively. PCR-RFLP was more discriminatory than auxotyping, which classifies the strains into only two auxotypes. Some strains belonging to common serovars could be further differentiated. A combination of PCR-RFLP analysis, auxotyping and serotyping further increased the discrimination of the strains into 34 subtypes. The PCR-RFLP method was easy to perform, reliable, reproducible, and consistent with published nucleotide sequence data. CONCLUSION: The PCR-RFLP method can augment auxotyping and serotyping or be used as a preliminary screening tool to differentiate N gonorrhoeae strains in areas where serotyping reagents are not easily available.

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Serotyping of Neisseria gonorrhoeae based on the differential recognition by a panel of six monoclonal antibodies (MAbs) against Protein IB (PIB) is a valuable tool for studying the epidemiology of gonorrhoea. However, the predominance of certain serovars in specific geographic regions necessitates the development of new MAbs or new genotyping approaches. Nucleotide and amino-acid sequence analysis of PIB from strains within the IB-3 and IB-7 serovars revealed strain differences within the same serovar. Based on the generation of PIB nucleotide and deduced amino-acid sequences that centred on amino-acid residues 196 and 237, eight serovar IB-3 strains and nine serovar IB-7 strains were separately subdivided into five groups. Intra-serovar differences were also established by pulsed-field gel electrophoresis (PFGE) of macro-restriction fragments generated by SpeI- and NheI-cleavage of genomic DNA. There was good correlation between the results based on PIB gene PCR-sequencing and those based on PFGE analysis of macro-restriction fragment patterns. These data demonstrate the high precision of PFGE analysis and indicate that this approach can be used as a rapid epidemiological subtyping system for major serovars of N. gonorrhoeae.

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A pair of primers were designed for the polymerase chain reaction (PCR) to amplify a 341-base pair fragment of the gene encoding the outer membrane protein IB (PIB) of Neisseria gonorrhoeae. This PCR technique is specific and sensitive, being able to detect gonococcal strains belonging to ten different PIB serovars, but not PIA gonococcus nor other negative control bacteria. PCR products of four representative PIB strains were directly sequenced. Of the three strains belonging to serovar IB4, two (S11 and S48) shared identical nucleotide and amino acid sequences in the PIB region examined. The third IB4 strain (S4) revealed sequences identical to the published IB26 strain (P9). The sequences of strains P9, S4, S11 and S48 were found to differ from those of strain S34 (serovar IB5). The PCR sequencing technique can further differentiate strains belonging to a common serovar and establish clonal relationships among strains. As a molecular epidemiological tool, the PCR-sequencing strategy can augment existing typing methods including serotyping.

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Genomic DNA from 48 Neisseria gonorrhoeae isolates was digested with low-frequency cleavage (LFC) endonucleases (SpeI and NheI) and analysed by contour-clamped homogeneous electric fields electrophoresis (CHEF). The restriction patterns generated were reproducible, stable, easy to read and offer a more practical approach than restriction endonuclease analysis (REA) with high-frequency cleavage (HFC) endonucleases. Strains sharing common auxotypes and serovars could be differentiated and correlation with auxotype/serovar (A/S) classes was demonstrated for some, but not all strains. The strains were distributed into 18 A/S classes and 38 SpeI and 40 NheI restriction patterns. This greater discriminatory power of CHEF REA should allow subdivision of strains within common A/S classes.

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