Multi-omics Analyses Reveal Synergistic Carbohydrate Metabolism in Streptococcus mutans-Candida albicans Mixed-Species Biofilms.

Candida albicans, a major opportunistic fungal pathogen, is frequently found together with Streptococcus mutans in dental biofilms associated with severe childhood caries (tooth decay), a prevalent pediatric oral disease. However, the impact of this cross-kingdom relationship on C. albicans remains largely uncharacterized. Here, we employed a novel quantitative proteomics approach in conjunction with transcriptomic profiling to unravel molecular pathways of C. albicans when cocultured with S. mutans in mixed biofilms. RNA sequencing and iTRAQ (isobaric tags for relative and absolute quantitation)-based quantitative proteomics revealed that C. albicans genes and proteins associated with carbohydrate metabolism were significantly enhanced, including sugar transport, aerobic respiration, pyruvate breakdown, and the glyoxylate cycle. Other C. albicans genes and proteins directly and indirectly related to cell morphogenesis and cell wall components such as mannan and glucan were also upregulated, indicating enhanced fungal activity in mixed-species biofilm. Further analyses revealed that S. mutans-derived exoenzyme glucosyltransferase B (GtfB), which binds to the fungal cell surface to promote coadhesion, can break down sucrose into glucose and fructose that can be readily metabolized by C. albicans, enhancing growth and acid production. Altogether, we identified key pathways used by C. albicans in the mixed biofilm, indicating an active fungal role in the sugar metabolism and environmental acidification (key virulence traits associated with caries onset) when interacting with S. mutans, and a new cross-feeding mechanism mediated by GtfB that enhances C. albicans carbohydrate utilization. In addition, we demonstrate that comprehensive transcriptomics and quantitative proteomics can be powerful tools to study microbial contributions which remain underexplored in cross-kingdom biofilms.

In vivo measurement of gadolinium diffusivity by dynamic contrast-enhanced MRI: a preclinical study of human xenografts.

Compartmental tracer kinetic models currently used for analysis of dynamic contrast-enhanced MRI data yield poor fittings or parameter values that are unphysiological in necrotic regions of the tumor, as these models only describe microcirculation in perfused tissue. In this study, we explore the use of Fick's law of diffusion as an alternative method for analysis of dynamic contrast-enhanced MRI data in the necrotic regions. Xenografts of various human cancer cell lines were implanted in 14 mice that were subjected to dynamic contrast-enhanced MRI performed using a spoiled gradient recalled sequence. Tracer concentration was estimated using the variable flip angle technique. Poorly perfused and necrotic tumor regions exhibiting delayed and slow enhancement were identified using a k-means clustering algorithm. Tracer behavior in necrotic regions was shown to be consistent with Fick's diffusion equation and the in vivo gadolinium diffusivity was estimated to be 2.08 (±0.88) × 10(-4) mm(2)/s. This study proposes the use of gadolinium diffusivity as an alternative parameter for quantifying tracer transport within necrotic tumor regions.

Diagnostic characteristics of clinical prediction models for obstructive sleep apnea in different clinic populations.

PURPOSE: As predictive factors and their diagnostic values are affected by the characteristics of the population studied, clinical prediction model for obstructive sleep apnea (OSA) may exhibit different diagnostic characteristics in different populations. We aimed to compare the diagnostic characteristics of clinical prediction models developed in two different populations. METHODS: One hundred seventeen consecutive clinic patients (group 1) were evaluated to develop a clinical prediction model for OSA (local model). The diagnostic characteristics of this local model were compared with those of a foreign model by applying both models to another group of 52 patients who were referred to the same clinic (group 2). All patients underwent overnight polysomnography. RESULTS: The local model had an area under receiver operator characteristics curve of 79%. A cutoff of 0.6 was associated with sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 77.9%, 72.5%, 84.5%, and 63.0%, respectively. The overall diagnostic accuracy, sensitivity, specificity, PPV, and NPV of the local model using data from patients in group 2 were 69.0%, 78.1%, 45.0%, 69.4%, and 56.3%, respectively. The foreign model had an overall diagnostic accuracy of 64.0% when applied to data from patients in group 2. At the optimal cutoff of 17, the foreign model was associated with sensitivity of 38.2%, specificity of 83.3%, NPV of 41.7% and PPV of 81.3%. CONCLUSIONS: Clinical prediction model for OSA derived from a foreign population exhibits markedly different diagnostic characteristics from one that is developed locally, even though the overall accuracy is similar. Our findings challenge the predictive usefulness and the external validity of clinical prediction models.

The impact of a delay in intensive care unit admission for community-acquired pneumonia.

The primary objective of the present study was to evaluate the effect on hospital mortality of a delay in intensive care unit (ICU) admission for severe community-acquired pneumonia (CAP). The secondary objectives were to assess if such delays were associated with treatment variations by the emergency department (ED) and deterioration in the general wards, and to evaluate the prognostic ability of the Infectious Disease Society of America (IDSA)/American Thoracic Society (ATS) minor criteria. We retrospectively compared patients who were admitted straight from the ED to the ICU (direct group, n = 54) and those who were first admitted from the ED to the general wards before ICU transfer (delayed group, n = 49), over 2.5 yrs. Even after excluding patients who required mechanical ventilation and/or vasopressors at the ED, delayed ICU admission was an independent predictor of hospital mortality (OR 9.61). The delayed group received fewer fluid boluses in the ED and rapidly deteriorated in the general wards. The presence of ≥3 IDSA/ATS minor criteria was associated with increased mortality in the delayed group. In conclusion, prompt recognition of severe CAP using the IDSA/ATS minor criteria, followed by aggressive management at the ED and direct ICU admission, are all crucial toward improving outcomes.

Effects of left bundle-branch block on cardiac structure, function, perfusion, and perfusion reserve: implications for myocardial contrast echocardiography versus radionuclide perfusion imaging for the detection of coronary artery disease.

BACKGROUND: We aimed to investigate the cardiac effects of left bundle-branch block (LBBB) using myocardial contrast echocardiography (MCE) to ascertain the value of MCE for detecting coronary artery disease (CAD) and to uncover the mechanism that affects the accuracy of single-photon emission computed tomography (SPECT) in these patients. METHODS AND RESULTS: Sixty-three symptomatic LBBB patients (group A), 10 left ventricular ejection fraction-matched control subjects without LBBB and no CAD (group B), and 10 normal control subjects (group C) underwent resting echocardiography. Rest and vasodilator MCE and SPECT were undertaken in LBBB patients. Septal (SW) and posterior wall (PW) thickness, thickening, quantitative myocardial blood flow (MBF), and MBF reserve were measured. SW/PW thickness and percentage thickening ratios were lower (P<0.01 and P<0.05, respectively) in group A compared with both groups B and C, but resting SW/PW MBF and MBF reserve ratios were similar in all 3 groups. MBF reserve but not MBF was reduced in groups A and B (2.2+/-0.7 versus 2.2+/-0.2; P=0.98) compared with group C (3.1+/-0.5; P<0.01). SW thickness was an independent predictor (P=0.006) of SPECT perfusion defects in LBBB patients without CAD. MCE (92%) had a sensitivity similar to SPECT (92%); however, the specificity of MCE (95%) was superior (P<0.0001) to SPECT (47%) for the detection of CAD. CONCLUSIONS: Despite asymmetrical reduction in SW thickness and function, MBF is preserved and MBF reserve is homogeneously reduced in LBBB patients with left ventricular systolic dysfunction. Because of partial volume effects, the accuracy of SPECT for detecting CAD was significantly compromised compared with MCE in this patient cohort.

Validation and clinical implications of the IDSA/ATS minor criteria for severe community-acquired pneumonia.

BACKGROUND: The 2007 Infectious Disease Society of America (IDSA)/American Thoracic Society (ATS) guidelines defined severe community-acquired pneumonia (CAP) and recommended intensive care unit (ICU) admission when patients fulfilled three out of nine minor criteria. These criteria have not been validated. METHODS: All patients admitted to our hospital from 2004 to 2007 for CAP were reviewed retrospectively. Patients who fulfilled any IDSA/ATS major criteria for severe CAP at the emergency department (ie, the need for mechanical ventilation or vasopressors) were excluded. The predictive characteristics of the IDSA/ATS minor criteria were compared with those of the Pneumonia Severity Index (PSI) and the CURB-65 score for hospital mortality and ICU admission. RESULTS: 1242 patients were studied (mean age 65.7 years, hospital mortality 14.7%). The areas under the receiver operating characteristic curves for the IDSA/ATS minor criteria were 0.88 (95% CI 0.86 to 0.91) and 0.85 (95% CI 0.81 to 0.88) for predicting hospital mortality and ICU admission, respectively. These were greater than the corresponding areas for the PSI and the CURB-65 score (p < 0.05). The sensitivity, specificity, positive and negative predictive values of the minor criteria were 81.4%, 82.9%, 45.2% and 96.3%, respectively, for hospital mortality and 58.3%, 90.6%, 52.9% and 92.3%, respectively, for ICU admission. The minor criteria were more specific than the PSI and more sensitive than the CURB-65 score for both outcomes. CONCLUSION: These findings support the use of the IDSA/ATS minor criteria to predict hospital mortality and guide ICU admission in inpatients with CAP who do not require emergency mechanical ventilation or vasopressors.

Metered-dose inhaler with spacer instead of nebulizer during the outbreak of severe acute respiratory syndrome in Singapore.

BACKGROUND: The use of metered-dose inhaler (MDI) with spacer instead of nebulizer may be important during an outbreak of an airborne infection. However, there is a paucity of data on patients' and nurses' abilities and perspectives on MDI with spacer for the treatment of acute airway obstruction during such an outbreak. METHODS: We evaluated 50 consecutive MDI-with-spacer treatments administered in the respiratory wards of the National University Hospital of Singapore, and interviewed the patients after each treatment during the outbreak of severe acute respiratory syndrome (SARS). We also conducted interviews with 50 nurses who had experience in administering bronchodilators via both nebulizer and MDI with spacer. RESULTS: Forty-six patients (92%) were able to use MDI with spacer effectively. Sixteen percent of the patients preferred nebulizer over MDI with spacer. Fifty-eight percent of the patients thought MDI with spacer was easier to use than nebulizer, and 34% thought MDI was as easy to use as nebulizer. Sixteen percent of the patients thought that nebulizer was more effective than MDI with spacer in relieving their symptoms. Ninety-six percent of the nurses preferred nebulizer over MDI with spacer. Forty-two nurses (84%) thought that nebulizer was more effective for treating acute airflow obstruction in the hospital. CONCLUSIONS: In the in-patient setting during an outbreak of an airborne infection, for treatment of acute airflow obstruction, MDI with spacer was acceptable and preferred by a high percentage of patients. However, a high percentage of nurses had misconceptions regarding the efficacy of and patients' ability to use MDI with spacer.

Use of traditional versus electronic medical-information resources by residents and interns.

BACKGROUND: Little is known about the information-seeking behaviour of junior doctors, with regard to their use of traditional versus electronic sources of information. AIMS: To evaluate the amount of time junior doctors spent using various medical-information resources and how useful they perceived these resources to be. METHODS: A questionnaire study of all residents and interns in a tertiary teaching hospital in July and August 2004. RESULTS: In total, 134 doctors returned the completed questionnaires (response rate 79.8%). They spent the most time using traditional resources like teaching sessions and print textbooks, rating them as most useful. However, electronic resources like MEDLINE, UpToDate, and online review articles also ranked highly. Original research articles were less popular. CONCLUSION: Residents and interns prefer traditional sources of medical information. Meanwhile, though some electronic resources are rated highly, more work is required to remove the barriers to evidence-based medicine.

Is a short arm cast appropriate for stable distal radius fractures in patients older than 55 years? A randomized prospective multicentre study.

We conducted a prospective randomized, multicentre study to compare short arm and long arm plaster casts for the treatment of stable distal radius fracture in patients older than 55 years. We randomly assigned patients over the age of 55 years who had stable distal radius fracture to either a short arm or long arm plaster cast at the first review 1 week after their injury. Radiographic and clinical follow-up was conducted at 1, 3, 5, 12 and 24 weeks following their injury. Also, degree of disability caused by each cast immobilization was evaluated at the patient's visit to remove the cast. There were no significant differences in radiological parameters between the groups except for volar tilt. Despite these differences in volar tilt, neither functional status as measured by the Disabilities of the Arm, Shoulder and Hand, nor visual analogue scale was significantly different between the groups. However, the mean score of disability caused by plaster cast immobilization and the incidence rate of shoulder pain were significantly higher in patients who had a long plaster cast. Our findings suggest that a short arm cast is as effective as a long arm cast for stable distal radius fractures in the elderly. Furthermore, it is more comfortable and introduces less restriction on daily activities. LEVEL OF EVIDENCE: II.

Dexmedetomidine-associated hyperthermia: a retrospective cohort study of intensive care unit admissions between 2009 and 2016.

Dexmedetomidine-associated hyperthermia has not been previously studied. Analysis is warranted to determine whether this potentially dangerous complication is more prevalent than previously realised. We aimed to examine the association between dexmedetomidine and temperature ≥39.5°C, including patient characteristics, temporality and potential risk factors. We conducted a retrospective cohort study of all intensive care unit (ICU) admissions between 1 July 2009 and 31 May 2016 in a tertiary ICU in Australia. Temperature data was available for 9,782 ICU admissions. Dexmedetomidine was given intravenously to 611 (6.3%) patients at a dose of 0 to 1.5 g/kg/hour. Temperatures ≥39.5°C were recorded in 341 (3.5%) patients. Overall hospital mortality was 10.8% for all admissions and 29.3% for patients with temperatures ≥39.5°C. Dexmedetomidine exposure was more frequent in patients with temperature recordings ≥39.5°C compared to those with temperatures <39.5°C, 11.94% versus 2.94% (odds ratio [OR] 4.49; 95% confidence intervals [CI] 3.37, 5.92; P <0.001). The association was stronger for patients post-open heart surgery (OHS) with temperatures ≥39.5°C (OR 12.9; 95% CI 5.01, 31.62; P <0.001). Multivariate analysis showed an independent association between dexmedetomidine and a temperature ≥39.5°C in two particular patient groups: OHS (OR 2.72; 95% CI 1.1, 6.9; P <0.001), and obesity (OR 3.44; 95% CI 1.5, 7.9; P <0.001). Dexmedetomidine exposure is associated with an increased risk of hyperthermia. Possible risk factors are open heart surgery and obesity.

Proteomic Profiling of De Novo Protein Synthesis in Starvation-Induced Autophagy Using Bioorthogonal Noncanonical Amino Acid Tagging.

Autophagy is an intracellular degradation process activated by stress factors such as nutrient starvation to maintain cellular homeostasis. There is emerging evidence demonstrating that de novo protein synthesis is involved in the autophagic process. However, up-to-date characterizing of these de novo proteins is technically difficult. In this chapter, we describe a novel method to identify newly synthesized proteins during starvation-mediated autophagy by bioorthogonal noncanonical amino acid tagging (BONCAT), in conjunction with isobaric tagging for relative and absolute quantification (iTRAQ)-based quantitative proteomics. l-azidohomoalanine (AHA) is an analog of methionine, and it can be readily incorporated into the newly synthesized proteins. The AHA-containing proteins can be enriched with avidin beads after a "click" reaction between alkyne-bearing biotin and the azide moiety of AHA. The enriched proteins are then subjected to iTRAQ™ labeling for protein identification and quantification using liquid chromatography-tandem mass spectrometry (LC-MS/MS). By using this technique, we have successfully profiled more than 700 proteins that are synthesized during starvation-induced autophagy. We believe that this approach is effective in identification of newly synthesized proteins in the process of autophagy and provides useful insights to the molecular mechanisms and biological functions of autophagy.

Prospective study of a new serological test (ASSURE TB Rapid Test) for the diagnosis of pulmonary tuberculosis.

OBJECTIVE: To prospectively compare a rapid tuberculosis serological test, ASSURE TB Rapid Test, with traditional smear and culture methods for the diagnosis of pulmonary tuberculosis (PTB). DESIGN: All consecutive in-patients aged > or = 18 years suspected of having active PTB and admitted between June 2001 and March 2003 were tested with three sputum samples for smear and culture of Mycobacterium tuberculosis and serology (done within 3 days). RESULTS: Of 238 patients initially enrolled (male: female 2.5:1, mean age 56.6 years), the final analysis included 216 patients. For the final diagnosis of PTB, the sensitivity and specificity of the serological test were respectively 60.2% (95%CI 50.5-69.1) and 82.3% (95%CI 74.2-88.2) compared to 53.4% (95%CI 43.8-62.7) and 98.2% (95%CI 93.8-99.5) for the smear test. A combination of smear and serology provided an increased sensitivity of 74.8% (95%CI 65.6-82.2), but a lower specificity of 80.5% (95%CI 72.3-86.8). CONCLUSION: The new serological test showed a moderate increase in sensitivity but a decrease in specificity compared to smear examination. The combination (smear + serology) test further increased the sensitivity while maintaining a moderate specificity.

Tuberculosis associated with Mycobacterium tuberculosis Beijing and non-Beijing genotypes: a clinical and immunological comparison.

BACKGROUND: The Mycobacterium tuberculosis Beijing genotype is biologically different from other genotypes. We aimed to clinically and immunologically compare human tuberculosis caused by Beijing and non-Beijing strains. METHODS: Pulmonary tuberculosis patients were prospectively enrolled and grouped by their M. tuberculosis genotypes. The clinical features, plasma cytokine levels, and cytokine gene expression levels in peripheral blood mononuclear cells (PBMC) were compared between the patients in Beijing and non-Beijing groups. RESULTS: Patients in the Beijing group were characterized by significantly lower frequency of fever (odds ratio, 0.12, p = 0.008) and pulmonary cavitation (odds ratio, 0.2, p = 0.049). Night sweats were also significantly less frequent by univariate analysis, and the duration of cough prior to diagnosis was longer in Beijing compared to non-Beijing groups (medians, 60 versus 30 days, p = 0.048). The plasma and gene expression levels of interferon (IFN) gamma and interleukin (IL)-18 were similar in the two groups. However, patients in the non-Beijing group had significantly increased IL-4 gene expression (p = 0.018) and lower IFN-gamma : IL-4 cDNA copy number ratios (p = 0.01). CONCLUSION: Patients with tuberculosis caused by Beijing strains appear to be less symptomatic than those who have disease caused by other strains. Th1 immune responses are similar in patients infected with Beijing and non-Beijing strains, but non-Beijing strains activate more Th2 immune responses compared with Beijing strains, as evidenced by increased IL-4 expression.

The clinical utility of Xpert® MTB/RIF testing in induced sputum.

We performed a prospective clinical audit in hospitalised patients with suspected pulmonary tuberculosis (PTB) with the objective of evaluating the accuracy and clinical utility of Xpert® MTB/RIF in induced sputum in an intermediate-burden setting. Of 450 patients audited, 61 (13.6%) were diagnosed with PTB based on positive culture. The sensitivity, specificity and positive and negative predictive values for Xpert in smear-negative cases for induced sputum were respectively 75%, 99.5%, 94.7% and 97.0%. Xpert in induced sputum may facilitate diagnostic yield and expedite treatment in up to a quarter of PTB patients.

Regulation of the biosynthesis of cytochrome P-450 in brewer's yeast. Role of cyclic AMP.

The drug metabolising enzyme cytochrome P-450 has been studied in great detail in mammalian systems and its presence in microorganisms is also well established. However, neither its function nor its means of control in brewer's yeast, Saccharomyces cerevisiae, has been investigated. We demonstrate here using yeast protoplasts that it is the intracellular concentration of cyclic AMP which controls, by repression, the de novo synthesis of the enzyme, and also that cyclic AMP concentrations are in turn inversely related to the concentration of glucose in the yeast growth medium.

Phase structures and transitions in fully hydrated diacyltrehalose.

Real time X-ray diffraction was used to examine the gel bilayer to disordered bilayer phase transition in fully hydrated dipalmitoyltrehalose. The L beta to L alpha phase transition was shown to proceed via a second-order thermodynamic process involving incommensurate mesophase bilayer repeat structures and the formation of an intermediate rectangular acyl chain packing subcell. This phenomenon has only been previously shown to occur for dihexadecylphosphatidylcholine (DHPC) and dipalmitoylphosphatidylcholine (DPPC) dihydrates undergoing stepwise (i.e., noncontinuous) temperature changes. It can thus be inferred that the presence of trehalose-trehalose intra-bilayer interactions is a sufficient condition to modify the acyl chain structural rearrangements within the bilayer as a function of temperature.

Noninvasive ventilation in hypercapnic acute respiratory failure due to chronic obstructive pulmonary disease vs. other conditions: effectiveness and predictors of failure.

OBJECTIVE: This study compared the effectiveness of noninvasive ventilation (NIV) and the risk factors for NIV failure in hypercapnic acute respiratory failure (ARF) due to chronic obstructive pulmonary disease (COPD) vs. non-COPD conditions. DESIGN AND SETTING: Prospective cohort study in the medical intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: 111 patients with hypercapnic ARF, 43 of whom had COPD exacerbations and 68 other conditions. Baseline characteristics of the two groups were similar. MEASUREMENTS AND RESULTS: The risk of NIV failure, defined as the need for endotracheal intubation, was significantly lower in COPD than in other conditions (19% vs. 47%). High APACHE II score was an independent predictor of NIV failure in COPD (OR 5.38 per 5 points). The presence of pneumonia (OR 5.63), high APACHE II score (OR 2.59 per 5 points), rapid heart rate (OR 1.22 per 5 beats/min), and high PaCO(2) 1 h after NIV (OR 1.22 per 5 mmHg) were independent predictors of NIV failure in the non-COPD group. Failure of NIV independently predicted mortality (OR 10.53). CONCLUSIONS: Noninvasive ventilation was more effective in preventing endotracheal intubation in hypercapnic ARF due to COPD than non-COPD conditions. High APACHE II score predicted NIV failure in both groups. Noninvasive ventilation was least effective in patients with hypercapnic ARF due to pneumonia.