Attention to time-of-day variability improves the reproducibility of gene expression patterns in multiple sclerosis.

Low reproducibility in gene expression profiles has been observed in transcriptome studies, and this often limits applying findings to clinical practice. Here, we show time-of-day effects on gene expression and analytical schemes to increase the reproducibility in expression patterns. We recruited patients with relapsing-remitting multiple sclerosis (RRMS) and healthy subjects and collected blood from individuals twice a day, day (2 pm) and night (9 pm). RNA sequencing analyses found that gene expression in RRMS in relapse (Relapse) is significantly changed at night compared with either Relapse at day or RRMS in remission (Remission). Gene set overrepresentation analysis demonstrated that gene sets significantly changed in Relapse at night are enriched to immune responses related to MS pathology. In those gene sets, 68 genes are significantly changed expression in Relapse at night compared with Relapse at day and Remission. This supports that times of sample collections should be standardized to obtain reproducible gene expression patterns.

High serum neurofilament levels among Chinese patients with aquaporin-4-IgG-seropositive neuromyelitis optica spectrum disorders.

BACKGROUND: Serum neurofilament light chain (sNfL) is a promising biomarker for neuromyelitis optica spectrum disorders (NMOSD) and multiple sclerosis (MS), but there is limited validation data in specific ethnic and disease groups. OBJECTIVE: To investigate the levels of sNfL in a cohort of Chinese patients with NMOSD and compare sNfL levels in patients with different disease courses and treatments. METHODS: We analysed sNfL levels in 153 Chinese patients with NMOSD (n = 51) and MS (n = 102) using single-molecule array (Simoa) technology. The sNfL levels were compared with those of 71 healthy controls from two centres in southern China. For each disease, we assessed correlations between sNfL and disease phases and treatments. RESULTS: Higher levels of sNfL were found in the patients with NMOSD [17.97 (10.55-27.94) pg/mL] and MS [15.83 (8.92-25.67) pg/mL] compared to healthy controls [10.09 (7.19-13.29) pg/mL, p < 0.001]. No significant differences were found between the AQP4-IgG-positive NMOSD group and OCB-positive MS group. CONCLUSIONS: sNfL measured by Simoa technology is a potential candidate blood biomarker for the diagnosis and disease monitoring of NMOSD in Chinese patients, warranting further prospective and multicentre studies.

Advanced MRI features in relapsing multiple sclerosis patients with and without CSF oligoclonal IgG bands.

Oligoclonal IgG bands (OCB) in cerebrospinal fluid (CSF) are important in diagnosis of multiple sclerosis (MS). We evaluated the MRI features of clinically definite MS subjects with and without CSF-OCB. Relapsing MS subjects were recruited from a prospective registry in a university center. CSF-OCB were detected using isoelectric focusing and lgG-specific immunofixation. MRI metrics including brain volumes, lesion volumes and microstructural measures, were analyzed by FMRIB Software Library (FSL) and Statistical Parametric Mapping (SPM). Seventy-five subjects with relapsing MS were analyzed. Forty-four (59%) subjects had an interval MRI at around 1 year. CSF-OCB were detected in 46 (61%) subjects. The OCB-positive group had a higher proportion of cerebellar lesions than the OCB-negative group (23.9% vs. 3.4%, p = 0.057). Except for amygdala volumes which were lower in the OCB-positive group (p = 0.034), other regional brain volumes including the subcortical deep gray matter and corpus callosum were similar. The two groups also showed comparable brain atrophy rate. For DTI, the OCB-positive group showed significantly higher mean diffusivity (MD) value in perilesional normal-appearing white matter (p = 0.043). Relapsing MS patients with and without CSF-OCB shared similar MRI features regarding volumetric analyses and DTI microstructural integrity.

Impaired cognition is related to microstructural integrity in relapsing remitting multiple sclerosis.

BACKGROUND: Cognitive impairment is common in multiple sclerosis (MS). However, the relationship between cognitive deficits and microstructural abnormalities in Chinese MS patients remains unclear. We aimed to investigate the importance of microstructural abnormalities and the associations with cognitive impairment in Chinese MS patients. METHODS: Three-dimensional T1-weighted magnetic resonance imaging (MRI) scans were obtained from 36 relapsing remitting MS patients. Diffusion tensor imaging (DTI) scans were acquired for 29 (81%) patients. Cognitive impairment was assessed using a comprehensive neuropsychological battery. Patients were classified into cognitively impaired (CI) group and cognitively preserved (CP) group. Using volBrain and FSL software, we assessed white matter lesion burden, white matter (WM) and gray matter (GM) volumetric as well as microstructural diffusivity. MRI variables explaining cognitive impairment were analyzed. RESULTS: Fifteen (42%) patients were classified as CI. Verbal learning and memory was the most commonly impaired domain (n = 16, 44%). CI patients had lower mean skeleton fractional anisotropy (FA) value than CP patients (275.45 vs. 283.61 × 10-3 , P = 0.023). The final predicting model including demographic variables and global skeleton mean diffusivity (MD) explained 43.6% of variance of the presence of cognitive impairment (ß = 0.131, P = 0.041). CI patients showed a widespread change of microstructural integrity comparing to CP patients, which was rarely overlapping with lesion probability map. Microstructural abnormalities in corpus callosum were associated with performance in verbal learning and memory, processing speed and selective attention (P < 0.05). CONCLUSION: Loss of microstructural integrity demonstrated by DTI helps explain cognitive dysfunction in Chinese MS patients.

Pneumocystis pneumonia in a patient treated with alemtuzumab for relapsing multiple sclerosis.

Pneumocystis jirovecii pneumonia (PJP) is a known risk in patients with chronic lymphocytic leukemia treated with alemtuzumab (Campath®). However, no recommendation for PJP prophylaxis for alemtuzumab use in multiple sclerosis (Lemtrada®) and no known associated PJP has been reported to date. We report a patient who developed PJP two months after receiving the first course of Lemtrada, and fully recovered after receiving cotrimoxazole treatment. We should remain vigilant of opportunistic infections in patients who develop pneumonitis and evaluate the need for PJP prophylaxis during Lemtrada treatment.

Comparison of multiple sclerosis patients with and without oligoclonal IgG bands in South China.

The data for oligoclonal IgG bands (OCB) in cerebrospinal fluid and the association with clinical profiles of Chinese patients with multiple sclerosis (MS) is lacking. We aimed to investigate the positive OCB incidence, as well as the clinical and magnetic resonance imaging (MRI) features associated with positive OCB in MS patients in South China. Consecutive MS patients were recruited from two centers, located in Guangzhou city and Hong Kong in South China. MS was re-diagnosed by McDonald criteria 2017. OCB was tested using isoelectric focusing method. The difference in the clinical and MRI features between OCB-positive and OCB-negative MS patients was evaluated. Among 184 MS patients analyzed (102 from Guangzhou and 82 from Hong Kong), 110 (59.8%) patients were OCB-positive. Except for onset age (P = 0.019), there was no significant difference between MS patients with or without OCB in relation to the course of disease, symptoms of first attack, lesion distribution on MRI, relapse rates, or disability progression. Our results demonstrate no significant clinical differences between the OCB positive and negative MS patients in this MS cohort. Larger cohort in Chinese patients with MS is warranted.

High prevalence and indexes of anti-John Cunningham virus antibodies in a cohort of Chinese patients with multiple sclerosis.

We performed a cross-sectional study in 123 Chinese multiple sclerosis patients residing in Hong Kong to evaluate their anti-John Cunningham virus status using STRATIFY JCV DxSelect assays. Anti-John Cunningham virus antibody was present in 98/123 (80%) subjects, among which 75/98 (77%) had an anti-John Cunningham virus index ≥1.5. Anti-John Cunningham virus antibody seropositivity was not correlated with age, disease duration, Expanded Disability Status Scale scores, types of multiple sclerosis (relapsing vs progressive), or disease-modifying treatments used. We found a very high seroprevalence and index of anti-John Cunningham virus antibodies in Chinese multiple sclerosis patients, which may impact the risk assessment and recommendation of disease-modifying treatments in this population.

Prevalence of neutralising antibodies to interferon-beta and clinical response in Chinese patients with relapsing multiple sclerosis.

BACKGROUND: There are no data on neutralising antibodies to interferon-beta and its clinical implications in Chinese patients with multiple sclerosis (MS). OBJECTIVES: The objectives of this study were to investigate the prevalence of neutralising antibodies among Chinese patients with relapsing MS receiving interferon-beta (1a or 1b) and to study the association between neutralising antibodies and the clinical-radiological response. METHODS: We performed a cross-sectional study on MS patients who received interferon-beta for 9 months or more, and evaluated the clinical response by relapses and magnetic resonance imaging lesions. Blood samples were evaluated for myxovirus resistance protein A (MxA) gene expression by polymerase chain reaction, anti-interferon-beta binding antibodies by enzyme-linked immunosorbent assay, and neutralising antibodies by cell-based MxA protein induction and luciferase reporter gene assays. Assay performances were evaluated by receiver operating characteristic analysis. RESULTS: Among 78 subjects recruited, 61/77 (79%) had anti-interferon-beta binding antibodies, and 22/78 (28%) had neutralising antibodies by MxA protein induction assay. The presence of high-titre neutralising antibodies was associated with poor clinical outcome (odds ratio 6.1, 95% confidence interval 1.5-25.6, P = 0.013). The sensitivity and specificity for neutralising antibodies using MxA gene expression assay (cut-off 0.20) was 80% and 68%, respectively (area under the curve 0.71). CONCLUSIONS: Neutralising antibodies are associated with poor clinical outcome in Chinese patients with relapsing MS. MxA gene expression and protein induction assays are complimentary assays for neutralising antibody detection.

Chronic kidney disease and the severity of coronary artery disease and retinal microvasculature changes: a cross-sectional study.

BACKGROUND: Prior studies have suggested the association between incidence of coronary artery disease (CAD) with chronic kidney disease (CKD) and poor glomerular filtration function. However to the best of our knowledge, few studies have specifically assessed this relationship based on the severity of CAD as quantified using Extent and Gensini scores. METHODS: Between June 2009 and January 2012, data were collected from 1,680 participants as part of the Australian Heart Eye Study (AHES) cohort. Coronary angiograms were scored according to Gensini (severity) and Extent scores. Retinal vessel calibres were measured using validated semi-automated software. Potential confounders were adjusted for using multivariate analysis. RESULTS: There were no significant associations found between CKD status with Extent and Gensini scores for CAD severity, both unadjusted and when adjusted for confounding factors. In unadjusted analyses, a significant association between CKD and narrower retinal arteriolar diameter was observed (P=0.0072). After multivariate adjustment, the association between CKD and retinal arteriolar diameter was attenuated and was no longer significant (P=0.1466). No associations were observed between retinal venular calibre and prevalent CKD. CONCLUSIONS: The present study demonstrated no independent associations between CKD and CAD severity. These results warrant validation by future large, prospective longitudinal studies.

Efficacy and cost-effectiveness of intracameral vancomycin in reducing postoperative endophthalmitis incidence in Australia.

BACKGROUND: Endophthalmitis is a rare but devastating postoperative complication of cataract surgery. We aimed to describe the incidence of acute postoperative endophthalmitis in an Australian population over a 14-year period. DESIGN: This was a retrospective longitudinal cohort study performed at Westmead Hospital, a major tertiary hospital in Sydney, Australia. PARTICIPANTS: Patients who had acute postoperative endophthalmitis within 6 weeks of their cataract surgery at Westmead Hospital were included. METHODS: Endophthalmitis cases from 2000 to 2014 were identified from computerized diagnostic coding. These were cross-referenced with the cataract surgery list over this time period. The routine use of intracameral vancomycin at the end of cataract surgery was introduced in Westmead Hospital in 2004. MAIN OUTCOME MEASURES: We quantified the incidence of acute postoperative endophthalmitis pre and post the routine use of intracameral vancomycin. RESULTS: A total of 14 805 cataract cases were performed at Westmead Hospital from 2000 to 2014. Seventeen cases of endophthalmitis were within 6 weeks post cataract surgery performed at Westmead Hospital. In the period 2000 to 2003, the incidence of postoperative endophthalmitis was 0.43% (11/2539). From 2004 to 2014, there was a dramatic decrease in the incidence of postoperative endophthalmitis to 0.049% (6/12 266, P < 0.0001). CONCLUSIONS: There has been a nine-fold reduction in the rate of acute postoperative endophthalmitis with the use of intracameral vancomycin in cataract surgery. Post-cataract surgery endophthalmitis is now a relatively rare cause of endophthalmitis in this Australian population. Our study supports the routine use of intracameral vancomycin as postoperative endophthalmitis prophylaxis.

Increase in the length of superficial temporal artery biopsy over 14 years.

BACKGROUND: Giant cell arteritis is a systemic inflammatory vasculitis of large-sized and medium-sized arteries. Superficial temporal artery biopsy of at least 20 mm has traditionally been the standard length for histopathology to accurately diagnose giant cell arteritis. Recent studies suggest than a post-fixation superficial temporal artery biopsy length of 7 to 10 mm is adequate for diagnosing giant cell arteritis. DESIGN: This is a retrospective observational study. PARTICIPANTS OR SAMPLES: The participants were all patients who underwent superficial temporal artery biopsy at Royal Prince Alfred Hospital, a large tertiary teaching hospital in Sydney, Australia, from 2008 to 2014. METHODS: Patients were identified using computerized hospital databases. Superficial temporal artery biopsy lengths were obtained from the histopathology reports. MAIN OUTCOME MEASURES: We aimed to compare the superficial temporal artery biopsy lengths performed at a large tertiary hospital over the past 7 years, to those performed from 2000 to 2005, and to determine the frequency of diagnosis of giant cell arteritis over the two time periods. RESULTS: There was a total of 96 superficial temporal artery biopsies performed from 2008 to 2014. The superficial temporal artery biopsy mean (standard deviation) length was 16.0(7.3) mm. This represented a significant (P = 0.015) increase in mean superficial temporal artery biopsy length when compared with a previous audit performed from 2000 to 2005 where the mean (standard deviation) superficial temporal artery biopsy was 11.7(6.2) mm. Of the 96 TABs, 20 (20.8%) were positive for giant cell arteritis, compared with a giant cell arteritis positivity rate of 20.4% for the previous audit period from 2000 to 2005. CONCLUSION: There has been a significant improvement in the length of superficial temporal artery biopsy performed at a tertiary hospital. Despite the increase in superficial temporal artery biopsy lengths, the giant cell arteritis positivity rate has remained stable.

Antenatal management of gestational diabetes mellitus can improve neonatal outcomes.

OBJECTIVE: Pregnancies complicated with gestational diabetes mellitus (GDM) are at a higher risk for caesarean and instrumental deliveries as well as adverse neonatal outcomes such as fetal overgrowth, hypoglycaemia and neonatal intensive care admission. Our primary objective was to describe neonatal outcomes in a sample that included term infants of both GDM mothers and mothers with normal glucose tolerance (NGT). DESIGN AND SETTING: this cross-sectional study included 599 term babies born between September and October 2010 at Royal Prince Alfred Hospital, Sydney, Australia. Maternal and neonatal data were collected from medical records and a questionnaire. Glycaemic control data was based on third trimester HbA1c levels and self-monitoring blood glucose levels (BGL). Univariate associations between GDM status and maternal demographic factors, as well as pregnancy outcomes, were estimated using χ(2) tests and t-tests, as appropriate. FINDINGS: of 599 babies, 67(11%) were born to GDM mothers. GDM mothers were more likely to be overweight/obese and of Asian ethnicity. Good glycaemic control was achieved in most GDM mothers. GDM babies were more likely to have been induced (p=0.013) and delivered earlier than non-GDM mothers (p<0.001), and they were also more likely to be breastfed within one hour of birth. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: in this study, GDM infants were more likely to be induced and delivered earlier but otherwise they did not have significantly different neonatal outcomes compared to infants of NGT mothers. This can be attributed to the good GDM control by lifestyle modification and insulin if necessary. The role of labour induction in GDM pregnancies should be further investigated. Midwives have an important role in maternal education during pregnancy and in the postnatal period.

Association between Retinal Arteriolar and Venule Calibre with Prevalent Heart Failure: A Cross-Sectional Study.

BACKGROUND: There is evidence to suggest that microvascular disease, particularly diabetic retinopathy, plays a role in the pathogenesis of HF. However, whether changes in retinal vessel calibre predicts HF is unclear. The purpose of this study was to examine the association of retinal microvascular structure with prevalent heart failure (HF). METHODS: The Australian Heart Eye Study (AHES) is a cross-sectional study that surveyed 1680 participants who presented to a tertiary referral hospital for the evaluation of potential coronary artery disease by coronary angiography. Retinal vessel calibre was graded using retinal photography and participants' self-reported echocardiography-confirmed HF was obtained via an extensive medical questionnaire. RESULTS: There were 107 participants (8.1%) with prevalent self-reported HF. Persons with wider retinal arteriolar calibre (comparing highest versus lowest tertile or reference) were more likely to have prevalent HF (OR 3.5; 95% CI, 1.7-7.2) when adjusted for age and sex. After further adjustment for body mass index, hypertension, diabetes, smoking status, triglycerides and estimated glomerular filtration rate, this association remained significant (OR 4.5; 95% CI, 2.0-9.8). After further stratification, this association remained significant among participants with diabetes (OR 10.3; 95% CI, 2.7-39.3) but not in those without diabetes (OR 2.7; 95% CI, 0.9-7.5). The strength of this association was not dependent on the length of history of diabetes, or retinopathy status. There was no significant association between retinal venular calibre and prevalence of HF. CONCLUSIONS: Wider retinal arteriolar diameter was significantly and independently associated with prevalent HF in participants of a cross-sectional study. This association was significant stronger among participants with diabetes compared to without diabetes. No association was found between retinal venule calibre with prevalent HF.

Platelets Play Differential Role During the Initiation and Progression of Autoimmune Neuroinflammation.

RATIONALE: Platelets are known to participate in vascular pathologies; however, their role in neuroinflammatory diseases, such as multiple sclerosis (MS), is unknown. Autoimmune CD4 T cells have been the main focus of studies of MS, although the factors that regulate T-cell differentiation toward pathogenic T helper-1/T helper-17 phenotypes are not completely understood. OBJECTIVE: We investigated the role of platelets in the modulation of CD4 T-cell functions in patients with MS and in mice with experimental autoimmune encephalitis, an animal model for MS. METHODS AND RESULTS: We found that early in MS and experimental autoimmune encephalitis, platelets degranulated and produced soluble factors serotonin (5-hydroxytryptamine), platelet factor 4, and platelet-activating factor, which specifically stimulated differentiation of T cells toward pathogenic T helper-1, T helper-17, and interferon-γ/interleukin-17-producing CD4 T cells. At the later stages of MS and experimental autoimmune encephalitis, platelets became exhausted in their ability to produce proinflammatory factors and stimulate CD4 T cells but substantially increased their ability to form aggregates with CD4 T cells. Formation of platelet-CD4 T-cell aggregates involved the interaction of CD62P on activated platelets with adhesion molecule CD166 on activated CD4 T cells, contributing to downmodulation of CD4 T-cell activation, proliferation, and production of interferon-γ. Blocking of formation of platelet-CD4 T-cell aggregates during progression of experimental autoimmune encephalitis substantially enhanced proliferation of CD4 T cells in the central nervous system and the periphery leading to exacerbation of the disease. CONCLUSION: Our study indicates differential roles for platelets in the regulation of functions of pathogenic CD4 T cells during initiation and progression of central nervous system autoimmune inflammation.

Patient experiences in retinal trials: a cross-sectional study.

BACKGROUND: Patient-centered care recognizes the obligation to understand and meet patient's expectations. An individual's satisfaction has been found to affect health-related decisions and treatment-related behaviours, which in turn affect medical compliance, follow-up, the success of treatment and the appropriate use of services. We studied the expectations, experiences and satisfaction of patients who participated in clinical trials for retinal diseases at the Sydney Eye Hospital. METHODS: The study was undertaken at the research clinic of the major public quaternary eye hospital in New South Wales, Australia. A 37-question survey was conducted on patients enrolled in or who had finished a clinical trial for macular disease in the 12 months preceding this study in November 2012. Patient satisfaction was assessed using close-ended, multiple choice questions. First, the decision making process for entering into the clinical trial was evaluated. Then the level of patient understanding and experience during the study was assessed. Finally, there was a series of questions to gauge the participants' perception of trial outcomes and overall impression gained from the experience. RESULTS: Eighty patients completed the questionnaire. Overall patient satisfaction was high with the majority of patients stating they would recommend participation in a retinal clinical trial (94 %) and participate in a subsequent trial (78 %). Most patients rated themselves as the most important factor in making the decision to join a trial. Patients felt well informed and expectations were generally felt to be met, however 14 % did not believe that they could withdraw from the study voluntarily. The most common reasons for trial participation were to contribute to medical science and to have improved treatment outcomes. CONCLUSIONS: We found that patients generally found participation in retinal clinical trials to be a positive experience. Factors contributing to dissatisfaction mainly related to inconvenience experienced by transportation and waiting times. We also found that patients felt well informed about the study, but some did not have a complete understanding of their rights, which had been communicated to them when they entered the study. There were both altruistic and self-motivated reasons behind patients' decisions to join a retinal trial.

Fetal and maternal factors associated with neonatal adiposity as measured by air displacement plethysmography: a large cross-sectional study.

BACKGROUND: There is evidence that the fetal and early postnatal environments play a role in determining the risk of lifetime obesity, diabetes and cardiovascular disease. Neonatal body composition, as a surrogate marker of the in-utero environment, can be reliably and accurately measured by air displacement plethysmography (ADP). Our primary objective was to identify preconception, fetal and maternal factors affecting neonatal body composition. METHODS: This cross-sectional study included 599 term babies born between September and October 2010 at Royal Prince Alfred Hospital, Sydney, Australia. Neonatal body fat percentage (BF%) was measured within 48 h of birth using ADP. Maternal demographic, anthropometric and medical data as well as neonatal gestational age and sex were used to develop a regression model that predicted body composition and birthweight. RESULTS: The mean (SD) neonatal BF% in our whole population was 9.2(4.4)%. Significant variables in the model for neonatal BF% were neonatal sex, gestational age, maternal ethnicity, gestational weight gain (GWG), pre-pregnancy BMI, parity and maternal hypertension (p<0.05); together, these explained 19% of the variation in BF%. GDM status was not a significant variable. Neonatal female sex, maternal Caucasian ethnicity and increased gestational weight gain explained the most variation and were most strongly associated with increased BF%. CONCLUSIONS: This study highlights maternal obesity and increased gestational weight gain as two factors that are amenable to intervention as risk factors for newborn adiposity, which is important in the future study of the "developmental origins of health and disease" hypothesis.