MiR-181a-5p promotes neural stem cell proliferation and enhances the learning and memory of aged mice.

Hippocampal neural stem cell (NSC) proliferation is known to decline with age, which is closely linked to learning and memory impairments. In the current study, we found that the expression level of miR-181a-5p was decreased in the hippocampal NSCs of aged mice and that exogenous overexpression of miR-181a-5p promoted NSC proliferation without affecting NSC differentiation into neurons and astrocytes. The mechanistic study revealed that phosphatase and tensin homolog (PTEN), a negative regulator of the AKT signaling pathway, was the target of miR-181a-5p and knockdown of PTEN could rescue the impairment of NSC proliferation caused by low miR-181a-5p levels. Moreover, overexpression of miR-181a-5p in the dentate gyrus enhanced the proliferation of NSCs and ameliorated learning and memory impairments in aged mice. Taken together, our findings indicated that miR-181a-5p played a functional role in NSC proliferation and aging-related, hippocampus-dependent learning and memory impairments.

Near infrared spectroscopy quantification based on Bi-LSTM and transfer learning for new scenarios.

This study proposed a deep transfer learning methodology based on an improved Bi-directional Long Short-Term Memory (Bi-LSTM) network for the first time to address the near infrared spectroscopy (NIR) model transfer issue between samples. We tested its effectiveness on two datasets of manure and polyglutamic acid (γ-PGA) solution, respectively. First, the optimal primary Bi-LSTM networks for cattle manure and the first batch of γ-PGA were developed by ablation experiments and both proved to outperform one-dimensional convolutional neural network (1D-CNN), Partial Least Square (PLS) and Extreme Learning Machine (ELM) models. Then, two types of transfer learning approaches were carried out to determine model transferability to non-homologous samples. For poultry manure and the second batch of γ-PGA, the obtained predicting results verified that the second approach of fine-tuning Bi-LSTM layers and re-training FC layers transcended the first approach of fixing Bi-LSTM layers and only re-training FC layers by reducing the RMSEPtarget of 23.4275% and 50.7343%, respectively. Finally, comparisons with fine-tuning 1D-CNN and other traditional model transfer methods further identified the superiority of the proposed methodology with exceeding accuracy and smaller variation, which decreased RMSEPtarget of poultry manure and the second batch of γ-PGA of 7.2832% and 48.1256%, 67.1117% and 80.6924% when compared to that acquired by fine-tuning 1D-CNN, Tradaboost-ELM and CCA-PLS which were the best of five traditional methods, respectively. The study demonstrates the potential of the Fine-tuning-Bi-LSTM enabled NIR technology to be used as a simple, cost effective and reliable detection tool for a wide range of applications under various new scenarios.

lncNBAT1/APOBEC3A is a mediator of HBX-induced chemoresistance in diffuse large B cell lymphoma cells.

Individuals with diffuse large B cell lymphoma (DLBCL) infected with hepatitis B virus (HBV) have worse chemotherapy efficacy and poorer outcomes. It is still unclear whether long noncoding RNAs (lncRNAs) serve as prognostic and therapeutic targets in the chemotherapy resistance of individuals with DLBCL and HBV infection. Here we found that the core component of HBV (HBX) directly upregulated the expression of lncNBAT1, which was closely associated with the chemotherapy outcomes of HBV-infected individuals with DLBCL. Upregulation of lncNBAT1 reduced the sensitivity of DLBCL cells to chemotherapeutic agents (methotrexate [MTX] or cytarabine [Ara-C]) that induced S phase arrest, whereas knockdown of lncNBAT1 significantly relieved the chemoresistance of HBX-expressing DLBCLs. Mechanistically, lncNBAT1 could interact with the signal transducer and activator of transcription 1 (STAT1) to prevent its enrichment at the promoter region of the functional target gene apolipoprotein B mRNA editing enzyme catalytic subunit 3A (APOBEC3A), inhibiting expression of APOBEC3A and inducing resistance to MTX in DLBCL cells. Furthermore, clinical data analysis showed that lncNBAT1 and APOBEC3A expression was closely related to the poor prognosis and short survival of individuals with DLBCL. Our findings suggest a potential prognostic marker and a candidate lncRNA target for treating HBV-infected individuals with DLBCL.

Mitochondrial genome characterization and phylogenetic analysis of arbuscular mycorrhizal fungus Rhizophagus sp.

In the present study, the complete mitochondrial genome of Rhizophagus sp. was assembled by the next-generation sequencing. We found that the complete mitochondrial genome of Rhizophagus sp. is 50,449 bp in length and consists of 14,741 (29.22%) adenine, 9427 (18.69%) cytosine, 9248 (18.33%) guanosine, and 17,033 (33.76%) thymine. The genome contains 24 conserved core protein-coding genes, 25 tRNA genes, and 2 rRNA genes. Phylogenetic analysis based on the combined mitochondrial gene set showed that Rhizophagus sp. has a close relationship with Rhizophagus fasciculatus, Glomus irregular, and G. intratadices.

Quantitative TLC-SERS detection of histamine in seafood with support vector machine analysis.

Scombroid fish poisoning caused by histamine intoxication is one of the most prevalent allergies associated with seafood consumption in the United States. Typical symptoms range from mild itching up to fatal cardiovascular collapse seen in anaphylaxis. In this paper, we demonstrate rapid, sensitive, and quantitative detection of histamine in both artificially spoiled tuna solution and real spoiled tuna samples using thin layer chromatography in tandem with surface-enhanced Raman scattering (TLC-SERS) sensing methods, enabled by machine learning analysis based on support vector regression (SVR) after feature extraction with principal component analysis (PCA). The TLC plates used herein, which were made from commercial food-grade diatomaceous earth, served simultaneously as the stationary phase to separate histamine from the blended tuna meat and as ultra-sensitive SERS substrates to enhance the detection limit. Using a simple drop cast method to dispense gold colloidal nanoparticles onto the diatomaceous earth plate, we were able to directly detect histamine concentration in artificially spoiled tuna solution down to 10 ppm. Based on the TLC-SERS spectral data of real tuna samples spoiled at room temperature for 0 to 48 hours, we used the PCA-SVR quantitative model to achieve superior predictive performance exceling traditional partial least squares regression (PLSR) method. This work proves that diatomaceous earth based TLC-SERS technique combined with machine-learning analysis is a cost-effective, reliable, and accurate approach for on-site detection and quantification of seafood allergen to enhance food safety.

Photonic Crystal-Enhanced Fluorescence Imaging Immunoassay for Cardiovascular Disease Biomarker Screening with Machine Learning Analysis.

When myocardial walls experience stress due to cardiovascular diseases, like heart failure, hormone N-terminal pro-B-type natriuretic peptide (NT-proBNP) is secreted into the blood. Early detection of NT-proBNP can assist diagnosis of heart failure and enable early medical intervention. A simple, cost-effective detection technique such as the widely used fluorescence imaging immunoassay is yet to be developed to detect clinically relevant levels of NT-proBNP. In this work, we demonstrate photonic crystal-enhanced fluorescence imaging immunoassay using diatom biosilica, which is capable of detecting low levels of NT-proBNP in solution with the concentration range of 0~100 pg/mL. By analyzing the fluorescence images in the spatial and spatial frequency domain with principle component analysis (PCA) and partial least squares regression (PLSR) algorithms, we create a predictive model that achieves great linearity with a validation R2 value of 0.86 and a predictive root mean square error of 14.47, allowing for good analyte quantification. To demonstrate the potential of the fluorescence immunoassay biosensor for clinical usage, we conducted qualitative screening of high and low concentrations of NT-proBNP in human plasma. A more advanced machine learning algorithm, the support vector machine classification, was paired with the PCA and trained by 160 fluorescence images. In the 40 testing images, we achieved excellent specificity of 93%, as well as decent accuracy and sensitivity of 78% and 65% respectively. Therefore, the photonic crystal-enhanced fluorescence imaging immunoassay reported in this article is feasible to screen clinically relevant levels of NT-proBNP in body fluid and evaluate the risk of heart failure.

Trace Detection of Tetrahydrocannabinol in Body Fluid via Surface-Enhanced Raman Scattering and Principal Component Analysis.

Tetrahydrocannabinol (THC) is the main active component in marijuana and the rapid detection of THC in human body fluid plays a critical role in forensic analysis and public health. Surface-enhanced Raman scattering (SERS) sensing has been increasingly used to detect illicit drugs; however, only limited SERS sensing results of THC in methanol solution have been reported, while its presence in body fluids, such as saliva or plasma, has yet to be investigated. In this article, we demonstrate the trace detection of THC in human plasma and saliva solution using a SERS-active substrate formed by in situ growth of silver nanoparticles (Ag NPs) on diatom frustules. THC at extremely low concentration of 1 pM in plasma and purified saliva solutions were adequately distinguished with good reproducibility. The SERS peak at 1603 cm-1 with standard deviation of 3.4 cm-1 was used for the evaluation of THC concentration in a methanol solution. Our SERS measurement also shows that this signature peak experiences a noticeable wavenumber shift and a slightly wider variation in the plasma and saliva solution. Additionally, we observed that THC in plasma or saliva samples produces a strong SERS peak at 1621 cm-1 due to the stretching mode of O-C═O, which is related to the metabolic change of THC structures in body fluid. To conduct a quantitative analysis, principal component analysis (PCA) was applied to analyze the SERS spectra of 1 pM THC in methanol solution, plasma, and purified saliva samples. The maximum variability of the first three principal components was achieved at 71%, which clearly denotes the impact of different biological background signals. Similarly, the SERS spectra of THC in raw saliva solution under various metabolic times were studied using PCA and 98% of the variability is accounted for in the first three principal components. The clear separation of samples measured at different THC resident times can provide time-dependent information on the THC metabolic process in body fluids. A linear regression model was used to estimate the metabolic rate of THC in raw saliva and the predicted metabolic time in the testing data set matched well with the training data set. In summary, the hybrid plasmonic-biosilica SERS substrate can achieve ultrasensitive, near-quantitative detection of trace levels of THC in complex body fluids, which can potentially transform forensic sensing techniques to detect marijuana abuse.

Molecular mechanisms of azole resistance in Candida bloodstream isolates.

BACKGROUND: Antifungal resistance rates are increasing. We investigated the mechanisms of azole resistance of Candida spp. bloodstream isolates obtained from a surveillance study conducted between 2012 and 2015. METHODS: Twenty-six azole non-susceptible Candida spp. clinical isolates were investigated. Antifungal susceptibilities were determined using the Sensititre YeastOne® YO10 panel. The ERG11 gene was amplified and sequenced to identify amino acid polymorphisms, while real-time PCR was utilised to investigate the expression levels of ERG11, CDR1, CDR2 and MDR1. RESULTS: Azole cross-resistance was detected in all except two isolates. Amino acid substitutions (A114S, Y257H, E266D, and V488I) were observed in all four C. albicans tested. Of the 17 C. tropicalis isolates, eight (47%) had ERG11 substitutions, of which concurrent observation of Y132F and S154F was the most common. A novel substitution (I166S) was detected in two of the five C. glabrata isolates. Expression levels of the various genes differed between the species but CDR1 and CDR2 overexpression appeared to be more prominent in C. glabrata. CONCLUSIONS: There was interplay of various different mechanisms, including mechanisms which were not studied here, responsible for azole resistance in Candida spp in our study.

Biological Photonic Crystal-Enhanced Plasmonic Mesocapsules: Approaching Single-Molecule Optofluidic-SERS Sensing.

Surface-enhanced Raman scattering (SERS) sensing in microfluidic devices, namely optofluidic-SERS, suffers an intrinsic trade-off between mass transport and hot spot density, both of which are required for ultra-sensitive detection. To overcome this compromise, photonic crystal-enhanced plasmonic mesocapsules are synthesized, utilizing diatom biosilica decorated with in-situ growth silver nanoparticles (Ag NPs). In our optofluidic-SERS testing, 100× higher enhancement factors and greater than 1,000× better detection limit were achieved compared with traditional colloidal Ag NPs, the improvement of which is attributed to unique properties of the mesocapsules. First, the porous diatom biosilica frustules serve as carrier capsules for high density Ag NPs that form high density plasmonic hot-spots. Second, the submicron-pores embedded in the frustule walls not only create a large surface-to-volume ratio allowing for effective analyte capture, but also enhance the local optical field through the photonic crystal effect. Last, the mesocapsules provide effective mixing with analytes as they are flowing inside the microfluidic channel. The reported mesocapsules achieved single molecule detection of Rhodamine 6G in microfluidic devices and were further utilized to detect 1 nM of benzene and chlorobenzene compounds in tap water with near real-time response, which successfully overcomes the constraint of traditional optofluidic sensing.

Quaternion-based Parallel Feature Extraction: Extending the Horizon of Quantitative Analysis using TLC-SERS Sensing.

Quantitative analysis using thin-layer chromatography coupled in tandem with surface-enhanced Raman scattering (TLC-SERS) still remains a grand challenge due to many uncontrollable variations during the TLC developing process and the random nature of the SERS substrates. Traditional chemometric methods solve this problem by sampling multiple SERS spectra in the sensing spot and then conducting statistical analysis of the SERS signals to mitigate the variation of quantitative analysis, while still ignoring the spatial distribution of the target species and the correlation among the multiple sampling points. In this paper, we proposed for the first time a parallel feature extraction and fusion method based on quaternion signal processing techniques, which can enable quantitative analysis using recently established TLC-SERS techniques. By marking three deterministic sampling points, we recorded spatially correlated SERS spectra to constitute an integral representation model of triple-spectra by a pure quaternion matrix. Quaternion principal component analysis (QPCA) was utilized for features extraction and followed by feature crossing among the quaternion principal components to obtain final fusion spectral feature vectors. Support vector regression (SVR) was then used to establish the quantitative model of melamine-contaminated milk samples with seven concentrations (1ppm to 250ppm). Compared with traditional TLC-SERS analysis methods, QPCA method significantly improved the accuracy of quantification by reaching only 7% and 2% quantization errors at 20 and 105 ppm concentration. Validation testing based on reasonable amount of statistic measurement results showed consistently smaller measurement errors and variance, which proved the effectiveness of QPCA method for TLC-SERS based quantitative sensing applications.

Correction to: Candidemia in a major regional tertiary referral hospital - epidemiology, practice patterns and outcomes.

[This corrects the article DOI: 10.1186/s13756-017-0184-1.].

Factors affecting clinical outcomes in the management of melioidosis in Singapore: a 16-year case series.

BACKGROUND: Burkholderia pseudomallei is a gram negative bacteria that causes a spectrum of human diseases in the tropics. Although melioidosis is endemic in Southeast Asia, large clinical case series were rarely reported from metropolitan Singapore. METHODS: This is a retrospective study of 219 consecutive patients with culture proven infections due to Burkholderia pseudomallei between the years 2001 to 2016 managed in Singapore General Hospital (SGH). We aimed to review local patients' characteristics and identify clinical factors associated with mortality and recurrent melioidosis. RESULTS: Culture proven melioidosis occurred in 219 patients, 83.1% were male with a mean age of 55.7 ± 14.3 years and 63.0% had diabetes mellitus. Most patients (71.7%) present within 4 weeks of symptom onset and the most common symptom was fever. The majority of patients had bacteremia (67.6%) and had infection involving the respiratory system (71.2%), presenting most frequently with multi-lobar pneumonia. Thirty-four (15.5%) deaths occurred during the initial hospitalisation with a median time from presentation to death of 6.0 days (interquartile range: 2.8-16.3). Twelve patients demised before the diagnosis of melioidosis was made. Univariate analysis identified patients with symptom duration of longer than 4 weeks, bacteremia, and disease requiring mechanical ventilation, inotropic support or temporary dialysis as factors that were significantly associated with mortality. Having bacteremia and disease requiring mechanical ventilation remained statistically significant factors in the multivariable analysis. Twenty-one (11.4%) patients developed at least 1 episode of culture proven recurrent infection, with 15 recurring within the first 12 months of their initial infection. Eight patients developed more than 1 episode of culture proven recurrent infection. Patients with multifocal infection were more likely to develop recurrent infection. CONCLUSION: In metropolitan Singapore, melioidosis was associated with mortality in excess of 15%, where more than a third occurred before diagnosis. This study reminds local physicians that melioidosis is still a serious infection affecting local male diabetic patients and an important differential diagnosis in a patient presenting with severe multi-lobar pneumonia and septic shock. Recurrent infections occurred in 11.4% and the weight-based dosing of oral eradication antibiotics may improve the management of this disease locally.

The Asia Cornea Society Infectious Keratitis Study: A Prospective Multicenter Study of Infectious Keratitis in Asia.

PURPOSE: To survey the demographics, risk factors, microbiology, and outcomes for infectious keratitis in Asia. DESIGN: Prospective, nonrandomized clinical study. METHODS: Thirteen study centers and 30 sub-centers recruited consecutive subjects over 12-18 months, and performed standardized data collection. A microbiological protocol standardized the processing and reporting of all isolates. Treatment of the infectious keratitis was decided by the managing ophthalmologist. Subjects were observed for up to 6 months. Main outcome measures were final visual acuity and the need for surgery during infection. RESULTS: A total of 6626 eyes of 6563 subjects were studied. The majority of subjects were male (n = 3992). Trauma (n = 2279, 34.7%) and contact lens wear (n = 704, 10.7%) were the commonest risk factors. Overall, bacterial keratitis was diagnosed in 2521 eyes (38.0%) and fungal keratitis in 2166 eyes (32.7%). Of the 2831 microorganisms isolated, the most common were Fusarium species (n = 518, 18.3%), Pseudomonas aeruginosa (n = 302, 10.7%), and Aspergillus flavus (n = 236, 8.3%). Cornea transplantation was performed in 628 eyes to manage ongoing infection, but 289 grafts (46%) had failed by the end of the study. Moderate visual impairment (Snellen vision less than 20/60) was documented in 3478 eyes (53.6%). CONCLUSION: Demographic and risk factors for infection vary by country, but infections occur predominantly in male subjects and are frequently related to trauma. Overall, a similar percentage of bacterial and fungal infections were diagnosed in this study. Visual recovery after infectious keratitis is guarded, and corneal transplantation for active infection is associated with a high failure rate.

Candidemia in a major regional tertiary referral hospital - epidemiology, practice patterns and outcomes.

BACKGROUND: Candidemia is a common cause of nosocomial bloodstream infections, resulting in high morbidity and mortality. This study was conducted to describe the epidemiology, species distribution, antifungal susceptibility patterns and outcomes of candidemia in a large regional tertiary referral hospital. METHODS: A retrospective surveillance study of patients with candidemia was conducted at Singapore General Hospital between July 2012 and December 2015. In addition, incidence densities and species distribution of candidemia episodes were analysed from 2008 to 2015. RESULTS: In the period of 2012 to 2015, 261 candidemia episodes were identified. The overall incidence was 0.14/1000 inpatient-days. C. glabrata (31.4%), C. tropicalis (29.9%), and C. albicans (23.8%) were most commonly isolated. The incidence of C. glabrata significantly increased from 2008 to 2015 (Coefficient 0.004, confidence interval 0-0.007, p = 0.04). Fluconazole resistance was detected primarily in C. tropicalis (16.7%) and C. glabrata (7.2%). fks mutations were identified in one C. albicans and one C. tropicalis. Candidemia episodes caused by C. tropicalis were more commonly encountered in patients with haematological malignancies (p = 0.01), neutropenia (p < 0.001) and higher SAPS II scores (p = 0.02), while prior exposure to echinocandins was associated with isolation of C. parapsilosis (p = 0.001). Echinocandins (73.3%) were most commonly prescribed as initial treatment. The median (range) time to initial treatment was 1 (0-9) days. The 30-day in-hospital mortality rate was 49.8%. High SAPS II score (Odds ratio, OR 1.08; 95% confidence interval, CI 1.05-1.11) and renal replacement therapy (OR 5.54; CI 2.80-10.97) were independent predictors of mortality, while drain placement (OR 0.44; CI 0.19-0.99) was protective. CONCLUSIONS: Decreasing azole susceptibilities to C. tropicalis and the emergence of echinocandin resistance suggest that susceptibility patterns may no longer be sufficiently predicted by speciation in our institution. Candidemia is associated with poor outcomes. Strategies optimising antifungal therapy, especially in the critically-ill population, should be explored.

[Quantitative analysis model of multi-component complex oil spill source based on near infrared spectroscopy].

Near infrared spectroscopy technology was used for quantitative analysis of the simulation of complex oil spill source. Three light petroleum products, i. e. gasoline, diesel fuel and kerosene oil, were selected and configured as simulated mixture of oil spill samples in accordance with different concentrations proportion, and their near infrared spectroscopy in the range of 8 000 -12 000 cm(-1) was collected by Fourier transform near infrared spectrometer. After processing the NIR spectra with different pretreatment methods, partial least squares method was used to establish quantitative analysis model for the mixture of oil spill samples. For gasoline, diesel fuel and kerosene oil, the second derivative method is the optimal pretreatment method, and for these three oil components in the ranges of 8 501.3-7 999.8 and 6 102.1-4 597.8 cm(-1); 6 549.5-4 597.8; 7 999.8-7 498.4 and 102.1-4 597.8 cm(-1), the correlation coefficients R2 of the prediction model are 0.998 2, 0.990 2 and 0.993 6 respectively, while the forecast RMSEP indicators are 0.474 7, 0.936 1 and 1.013 1 respectively; The experimental results show that using near infrared spectroscopy can quantitatively determine the content of each component in the simulated mixed oil spill samples, thus this method can provide effective means for the quantitative detection and analysis of complex marine oil spill source.

[Identification of spilled oil by NIR spectroscopy technology based on sparse nonnegative matrix factorization and support vector machine].

A novel method was proposed to discriminate different kinds of spilled oil. The identification of the spilled oils has great significance to developing the treatment program and tracking the source. The present method adapts to Fourier transform NIR spectrophotometer to collect the spectral data of simulation gasoline, diesel fuel and kerosene oil spills. The Sparse Nonnegative Matrix Factorization algorithm was used to extract features. Through training with 210 samples and 5-fold cross-validation, the authors constructed the qualitatvie analysis model based on support vector machine. The authors also researched the effect of the number of features and sparseness factor. The proposed method has the identification capabilities with the accuracy of 97.783 for 90 samples for validation. The present method of SNMF-SVM has a good identification effect and strong generalization ability, and can work as a new method for rapid identification of spilled oil.

Risk factors for contact lens-related fusarium keratitis: a case-control study in Singapore.

OBJECTIVE: To evaluate the specific contact lens-related or other factors that may contribute to the outbreak of Fusarium keratitis. METHODS: A case-control study was conducted of Fusarium keratitis in contact lens users in Singapore from March 1, 2005, to May 31, 2006, and included 61 patients with Fusarium keratitis and 188 population-based and 179 hospital-based control subjects. Interviewers asked about contact lens solution use and other risk factors. RESULTS: Patients with Fusarium keratitis were more likely to use ReNu contact lens solutions (Bausch & Lomb, Rochester, NY) 58 [95.1%] of 61 cases) than were either population-based (62 [34.3%] of 181) or hospital-based (50 [30.1%] of 166) control subjects. After controlling for age, sex, contact lens hygiene, and other factors, the use of ReNu with MoistureLoc significantly increased the risk of Fusarium keratitis (odds ratio, 99.3; 95% confidence interval, 18.4-535.4; P<.001), and the risk was 5 times higher compared with the risk with use of ReNu MultiPlus, a multipurpose solution (odds ratio, 21.5; 95% confidence interval, 4.0-115.5; P<.001). CONCLUSIONS: The use of ReNu contact lens solutions significantly increased the risk of contact lens-related Fusarium keratitis in Singapore. Our data support the recall of ReNu MultiPlus from the Singapore market and the need for further investigations into the role of ReNu MultiPlus in the development of Fusarium keratitis in other populations.

An outbreak of Fusarium keratitis associated with contact lens wear in Singapore.

CONTEXT: Fungal keratitis is a potentially blinding condition that is rarely seen with contact lens wear. OBJECTIVE: To describe a nationwide outbreak of fungal keratitis caused by Fusarium species among contact lens wearers in Singapore. DESIGN, SETTING, AND PATIENTS: Nationwide, hospital-based case series. All cases of fungal keratitis among contact lens wearers in all ophthalmology departments in Singapore were reviewed along with the charts of all contact lens wearers with culture-proven fungal keratitis from March 2005 through May 2006. A standardized telephone interview was conducted to obtain additional clinical information. MAIN OUTCOME MEASURE: Diagnosis of Fusarium keratitis associated with contact lens wear. RESULTS: During the study period, 66 patients (68 affected eyes) were diagnosed with Fusarium keratitis associated with contact lens wear; the estimated annual national incidence is 2.35 cases per 10,000 contact lens wearers (95% confidence interval, 0.62-7.22). Patients ranged in age from 13 to 44 years (mean [SD], 27.1 [8.4] years), of which 32 (48.5%) were men. The vast majority (65 patients; 98.5%) wore soft, disposable contact lenses; 62 patients (93.9%) reported using 1 brand of contact lens cleaning solution (ReNu, Bausch & Lomb, Rochester, NY), including 42 patients (63.6%) who recalled using ReNu with MoistureLoc. Most patients (81.8%) reported poor contact lens hygiene practices, including overnight use of daily wear contact lenses (19.7%), and use of contact lenses past the replacement date (43.9%). The final best-corrected visual acuity ranged from 20/20 to 20/80. Five patients (5 eyes; 7.4%) required emergency therapeutic or tectonic corneal transplantation. CONCLUSIONS: A new and evolving epidemic of Fusarium keratitis associated with contact lens wear was found in Singapore. Physicians and eye care practitioners worldwide need to be aware of the likelihood of similar outbreaks emerging among contact lens wearers.

Molecular typing of Salmonella enterica serovar typhi isolates from various countries in Asia by a multiplex PCR assay on variable-number tandem repeats.

A multiplex PCR method incorporating primers flanking three variable-number tandem repeat (VNTR) loci (arbitrarily labeled TR1, TR2, and TR3) in the CT18 strain of Salmonella enterica serovar Typhi has been developed for molecular typing of S. enterica serovar Typhi clinical isolates from several Asian countries, including Singapore, Indonesia, India, Bangladesh, Malaysia, and Nepal. We have demonstrated that the multiplex PCR could be performed on crude cell lysates and that the VNTR banding profiles produced could be easily analyzed by visual inspection after conventional agarose gel electrophoresis. The assay was highly discriminative in identifying 49 distinct VNTR profiles among 59 individual isolates. A high level of VNTR profile heterogeneity was observed in isolates from within the same country and among countries. These VNTR profiles remained stable after the strains were passaged extensively under routine laboratory culture conditions. In contrast to the S. enterica serovar Typhi isolates, an absence of TR3 amplicons and a lack of length polymorphisms in TR1 and TR2 amplicons were observed for other S. enterica serovars, such as Salmonella enterica serovar Typhimurium, Salmonella enterica serovar Enteritidis, and Salmonella enterica serovar Paratyphi A, B, and C. DNA sequencing of the amplified VNTR regions substantiated these results, suggesting the high stability of the multiplex PCR assay. The multiplex-PCR-based VNTR profiling developed in this study provides a simple, rapid, reproducible, and high-resolution molecular tool for the epidemiological analysis of S. enterica serovar Typhi strains.