Stimulation of monocytes, macrophages, and microglia by amphotericin B and macrophage colony-stimulating factor promotes remyelination.

Approaches to stimulate remyelination may lead to recovery from demyelinating injuries and protect axons. One such strategy is the activation of immune cells with clinically used medications, since a properly directed inflammatory response can have healing properties through mechanisms such as the provision of growth factors and the removal of cellular debris. We previously reported that the antifungal medication amphotericin B is an activator of circulating monocytes, and their tissue-infiltrated counterparts and macrophages, and of microglia within the CNS. Here, we describe that amphotericin B activates these cells through engaging MyD88/TRIF signaling. When mice were subjected to lysolecithin-induced demyelination of the spinal cord, systemic injections of nontoxic doses of amphotericin B and another activator, macrophage colony-stimulating factor (MCSF), further elevated the representation of microglia/macrophages at the site of injury. Treatment with amphotericin B, particularly in combination with MCSF, increased the number of oligodendrocyte precursor cells and promoted remyelination within lesions; these pro-regenerative effects were mitigated in mice treated with clodronate liposomes to reduce circulating monocytes and tissue-infiltrated macrophages. Our results have identified candidates among currently used medications as potential therapies for the repair of myelin.

Reduction of microglial activity in a model of multiple sclerosis by dipyridamole.

BACKGROUND: Despite extensive and persistent activation of microglia in multiple sclerosis (MS), microglia inhibitors have not yet been identified for treatment of the disorder. We sought to identify medications already in clinical use that could inhibit the activation of microglia. On the basis of the reported inhibitory effects of dipyridamole on phosphodiesterase activity that result in the production of various anti-inflammatory outcomes, we selected it for study. Dipyridamole is used clinically for secondary prevention in stroke. In this study, dipyridamole was examined using microglia in culture and in the mouse model of MS, experimental autoimmune encephalomyelitis (EAE). RESULTS: We found that dipyridamole attenuated the elevation of several cytokines and chemokines in human microglia caused by Toll-like receptor stimulation. Morphological characteristics of activated microglia in culture were also normalized by dipyridamole. In mice, dipyridamole decreased the clinical severity of EAE and reduced microglial activity and other histological indices of EAE in the spinal cord. CONCLUSIONS: Dipyridamole is an inhibitor of microglia activation and may have a role in MS and other neurological conditions to attenuate microglial activity.

Chondroitin sulfate proteoglycans in demyelinated lesions impair remyelination.

OBJECTIVE: Failure of remyelination is a critical impediment to recovery in multiple sclerosis (MS). Chondroitin sulfate proteoglycans (CSPGs) have been reported to accumulate in MS lesions, and we thus examined the functional roles of CSPGs on oligodendrocyte precursor cells (OPCs), oligodendrocytes, and remyelination. METHODS: We evaluated the expression of CSPGs in lysolecithin-injected mouse spinal cord, an animal model of demyelination and spontaneous remyelination. The functional impact of CSPGs on OPCs and remyelination was investigated using cultured adult murine and human OPCs and by treating demyelinated mice with xyloside to reduce the CSPG deposition that occurred following injury. RESULTS: Early and robust upregulation of CSPGs following lysolecithin-induced demyelination was cleared during remyelination. In culture, CSPGs anchored onto the substratum reduced the adhesion of mouse and human OPCs and their subsequent morphological differentiation into process-bearing oligodendrocytes. Soluble CSPGs added to already adherent OPCs reduced the development of processes, whereas the acquisition of mature myelin proteins was unimpeded. Stripe assays of alternating CSPG and control substrata confirmed the nonpermissive nature of CSPGs for OPC adhesion and morphological differentiation. Enzymatic degradation of CSPGs with chondroitinase ABC was sufficient to overcome CSPG-dependent inhibition of human oligodendrocytes. Finally, in vivo xyloside treatment to reduce CSPG synthesis in lysolecithin-demyelinated mice increased numbers of OPCs and oligodendrocytes in lesions, and culminated in improved remyelination. INTERPRETATION: These results identify CSPGs as a nonpermissive substrate for OPCs and oligodendrocytes, and as a prominent impediment to remyelination. The data suggest the requirement for the neutralization of CSPGs for repair after demyelination.

Structured approach to patients with memory difficulties in family practice.

OBJECTIVE: To provide family physicians with a structured approach to patients presenting with memory difficulties. SOURCES OF INFORMATION: The approach is based on an accredited memory clinic training program developed by the Centre for Family Medicine Memory Clinic in partnership with the Ontario College of Family Physicians. MAIN MESSAGE: Use of a structured clinical reasoning approach can assist physicians in achieving an accurate diagnosis in patients presenting with memory difficulties. Delirium, depression, and reversible causes need to be excluded, followed by differentiation among normal cognitive aging, mild cognitive impairment, and dementia. Obtaining collateral history and accurate functional assessment are critical. Common forms of dementia can be clinically differentiated by the order in which symptoms appear and by how cognitive deficits evolve over time. Typically, early signs of Alzheimer dementia involve impairment in episodic memory, whereas dementia involving predominantly vascular causes might present with early loss of executive function and relatively preserved episodic memory. Frontotemporal dementia and Lewy body spectrum disorders might have early loss of executive function and visuospatial function, as well as characteristic clinical features. CONCLUSION: A clinical reasoning approach can help physicians achieve early, accurate diagnoses that can guide appropriate management and improve care for patients with memory difficulties.

The use of weather nowcasting convolutional neural network extrapolators in cardiac PET imaging.

INTRODUCTION: Algorithms to predict short-term changes in local weather modalities have been used in meteorology for many years. These algorithms predict the temporospatial change in the movement of weather patterns such as cloud cover or precipitation. This paper extends convolutional neural network models for weather prediction/nowcasting to predict evolution in the extrapolation of sequentially acquired count data seen with cardiac positron tomography (PET) data to expected value temporally rather than spatially. METHODS: Six different algorithms used for nowcasting were modified and applied to confirm the approach. These algorithms were trained on an image data set of both simulated ellipsoids and simulated cardiac PET data. The peak signal-to-noise ratio (PSNR) and structural similarity (SSIM) were calculated for each of these trained models. They were compared to the BM3D denoising algorithm as a baseline comparison to a standard method of image denoising. RESULTS: Most of the implemented algorithms showed a significant improvement in both PSNR and SSIM when compared with the baseline standard, especially when the algorithms were implemented in combination. The best results were obtained with a combination of the ConvLSTM and TrajGRU algorithms with a PSNR improvement over the standard of 5 and more than double the SSIM metric. CONCLUSION: This approach of using serially acquired count data to extrapolate a future expected representation through convolutional neural networks has been shown to produce accurate representations of the expected value when compared with a baseline analytic methodology. This paper confirms that algorithms such as these can be used to substantially improve image estimation and shows significant improvement over a baseline standard.

Predominance of Th2 polarization by vitamin D through a STAT6-dependent mechanism.

BACKGROUND: Vitamin D has several reported immunomodulatory properties including the reduced generation of pro-inflammatory CD4+ T helper 1 (Th1) cells and the increase in levels of the anti-inflammatory Th2 subset. Less clear has been the impact of vitamin D on the pro-inflammatory Th17 subset, and whether and how vitamin D may preferentially drive the polarization of one of the T helper subsets. METHODS: Using human peripheral blood-derived mononuclear cells and mouse splenocytes and lymph node cells in culture, we examined whether and how vitamin D preferentially skews T cells towards the Th1, Th2 or Th17 subsets. Mice afflicted with the multiple sclerosis-like condition, experimental autoimmune encephalomyelitis (EAE), were examined in vivo for the relevance of the tissue culture-derived results. RESULTS: We report that the biologically active form of vitamin D, 1,25-dihydroxyvitamin D3 {1,25(OH)2D3}, consistently generates human and murine Th2 cells in culture, frequently leaving unchanged the levels of Th1/Th17 cytokines. As a result, the ratio of Th2 to Th1 and Th17 is increased by 1,25(OH)2D3. The upregulation of Th2 to Th1 or Th17 subsets by 1,25(OH)2D3 is enabled by an increase of the GATA-3 transcription factor, which itself is promoted upstream by an elevation of the STAT6 transcription factor. In mice, the alleviation of EAE severity by 1,25(OH)2D3 is accompanied by elevation of levels of GATA-3 and STAT6. Significantly, the efficacy of 1,25(OH)2D3 in ameliorating EAE is completely lost in mice genetically deficient for STAT6, which was accompanied by the inability of 1,25(OH)2D3 to raise GATA-3 in STAT6 null lymphocytes. CONCLUSIONS: These results of vitamin D promoting a Th2 shift through upstream GATA-3 and STAT6 transcription factors shed mechanistic understanding on the utility of vitamin D in MS.

A quantitative analysis of suspected environmental causes of MS.

BACKGROUND: Multiple sclerosis (MS) is a disease with purported environmental causes. Consistent correlations have been found in various settings for latitude, smoking exposure, sunlight, and vitamin D deficiency. We analysed the contribution of various environmental factors to the risk of developing MS from a population perspective. METHODS: We collated global data of MS prevalence from 54 studies over the previous ten years and calculated the degree of risk contributed by latitude, longitude, ultraviolet radiation (from NASA satellite data and formulae for available sunlight hours), population smoking rates (from WHO data), gender, study date, study demographics, and several socioeconomic factors. We report a very significant negative correlation between MS prevalence and available ultraviolet (UV) radiation. RESULTS: The lack of available UV radiation outweighs other factors by at least 20 fold (p < 10⁻8) from single variate regression analysis. Multiple regression analysis revealed that latitude and longitude are also significant factors; smoking may also provide a very minimal role. The eight prevalence studies from Scandinavia produced prevalences that were lower than expected, given their global geospatial positioning. CONCLUSIONS: The available ultraviolet radiation is a significant environmental factor, more so than all the other factors examined.

Seasonal variation of maternal serum vitamin D in Newfoundland and Labrador.

BACKGROUND: Research has suggested that vitamin D insufficiency and deficiency is common at northern latitudes, and that vitamin D insufficiency and deficiency may be common during pregnancy. We measured the serum 25-hydroxyvitamin D (25-[OH]D) status of pregnant women across the province of Newfoundland and Labrador in both summer and winter to investigate seasonal differences, age associations, and differences in geospatial distribution across the province. METHODS: We uniformly and randomly sampled blood from pregnant women in each of 79 census consolidated subdivisions across Newfoundland and Labrador from January to March 2007 and from July to September 2007. RESULTS: We obtained 304 samples from the end of winter (March) and 289 samples from the end of summer (September). Mean serum 25-(OH)D concentration was 52.1 nmol/L in winter and 68.6 nmol/L in summer (P < 0.001); 89% were vitamin D insufficient in the winter and 64% in the summer (P < 0.001); 6.6% were vitamin D deficient in winter and 1.7% in summer (P = 0.003), and younger women tended to be more vitamin D insufficient in the winter than older women. The geospatial distribution of vitamin D insufficiency tends to follow a north-south distribution in the winter. CONCLUSIONS: A significant proportion of pregnant women in Newfoundland and Labrador are vitamin D insufficient. Vitamin D insufficiency may have important adverse health consequences for both the mother and the fetus. Further study is necessary to address health outcomes and effects of vitamin D supplementation and lifestyle changes in this population.

Vitamin D insufficiency common in newborns, children and pregnant women living in Newfoundland and Labrador, Canada.

Vitamin D deficiency is associated with poor bone health, colorectal cancer, type 1 diabetes and multiple sclerosis. Two national health-related societies in Canada have made recommendations for vitamin D supplementation, yet little research has been reported on the vitamin D status of Canadians. Lifestyle changes, such as sunscreen use, spending less time outdoors and insufficient intake of vitamin D-containing foods as well as northern latitude, may be affecting human vitamin D status. A cross-sectional analysis of 25-hydroxyvitamin D [25-(OH)D] was conducted in pregnant women, newborns (umbilical cord blood) and children. Samples were analysed by liquid chromatography mass spectrometry. Published ranges for 25-(OH)D were used to determine vitamin D status. The prevalence of 25-(OH)D deficiency for the three groups studied revealed most concentrations in the 25-(OH)D deficiency or insufficiency ranges. There were significant differences in all groups studied between seasons, with the exception of maternal blood and female cord blood samples. 25-(OH)D insufficiency was common in all groups for winter and summer, more so in winter. 25-(OH)D insufficiency was common in the three groups studied. The Newfoundland and Labrador population may be at increased risk for vitamin D insufficiency because of factors such as northern latitude and lifestyle issues. Further research on the vitamin D status of this population is important, considering the potential adverse health-related outcomes and the recommendations on supplementation being made.

Time series analysis of ultraviolet B radiation and type 1 diabetes in Newfoundland.

BACKGROUND: Type 1 diabetes mellitus (T1DM) has been previously been associated with decreased levels of vitamin D. This study investigates the temporal association between average daily ultraviolet B (UVB) irradiance and T1DM in Newfoundland. METHODS: A complete list of patients diagnosed with T1DM in the province of Newfoundland and Labrador was constructed using multiple sources. Pooled and unpooled monthly incidence data along with monthly UVB measurements were used to build a time series transfer function model. The model was used to predict the future incidence of T1DM based on previous monthly trends, and these predictions were compared with actual measured incidences. RESULTS: A seasonal variation in pooled monthly incidence was observed. The transfer function model was able to reasonably predict the future incidence of T1DM based on previous observations and monthly UVB measurements. Tests of seasonality demonstrated a significant seasonal trend (p = 0.0003). CONCLUSIONS: This study suggests that erythemal UVB radiation may be temporally associated with the incidence of T1DM.

1,25-Dihydroxyvitamin D3 Protects against Immune-Mediated Killing of Neurons in Culture and in Experimental Autoimmune Encephalomyelitis.

Several studies have reported that low vitamin D levels are associated with an increased risk of developing multiple sclerosis (MS). As MS is an inflammatory disorder with degeneration of axons and neurons, we examined whether the biologically active form of vitamin D, 1,25-dihydroxyvitamin D3 (1,25D3), could protect against the T cell-mediated killing of human neurons in culture, and the axonal loss seen in mice with experimental autoimmune encephalomyelitis (EAE). Human neurons were exposed to activated human T lymphocytes and the loss of neurons was documented 24 hours later by counting the number of microtubule-associated protein-2 positive cells. Mice with EAE were harvested for counts of axonal profiles in the spinal cord. 1,25D3 was exposed to T cells in culture or administered to mice from peak EAE clinical severity when axonal loss was already evolving. Activated T lymphocytes killed human neurons prominently within 24 hours but toxicity was significantly attenuated when T cells were exposed to 1,25D3 prior to the co-culture. In EAE, 1,25D3 treatment initiated from peak clinical severity reduced the extent of clinical disability and mitigated the progressive loss of axons. The reduction of axonal and neuronal loss by 1,25D3 in the context of an inflammatory assault to the central nervous system is a potential contributor to the putative benefits of vitamin D in MS.

Laquinimod reduces neuroaxonal injury through inhibiting microglial activation.

OBJECTIVE: Laquinimod is an emerging oral medication for multiple sclerosis (MS) that reduces brain atrophy and progression of disability in two Phase III clinical trials. The mechanism of these effects is unclear. Persistent activation of microglia occurs in MS and contributes to injury. Thus, we investigated whether laquinimod alters properties of microglia in culture and in experimental autoimmune encephalomyelitis (EAE), and whether this reduces neurodegeneration. METHODS: Microglia were cultured from human brains. EAE was induced in mice. RESULTS: The activation of human microglia increased levels of several pro- and anti-inflammatory cytokines and these elevations were attenuated by pretreatment with laquinimod. Laquinimod prevented the decline in activated microglia of miR124a, a microRNA implicated in maintaining microglia quiescence, and reduced the activity of several signaling pathways (Jun-N-terminal kinase, ribosomal S6 kinase, and AKT/protein kinase B) in activated microglia. In EAE, axonal injury correlated with accumulation of microglia/macrophages in the spinal cord. EAE mice treated with laquinimod before onset of clinical signs subsequently had reduced microglia/macrophage density and axonal injury. Remarkably, when laquinimod treatment was initiated well into the disease course, the progressive demyelination, and axonal loss was halted. Besides inflammatory molecules associated with microglia, the level of inducible nitric oxide (NO) synthase capable of producing free radical toxicity was attenuated by laquinimod in EAE mice. Finally, in coculture where microglia activation caused neuronal death, laquinimod decreased NO levels, and neurotoxicity. INTERPRETATION: Laquinimod is a novel inhibitor of microglial activation that lowers microglia-induced neuronal death in culture and axonal injury/loss in EAE.

The geospatial relation between UV solar radiation and type 1 diabetes in Newfoundland.

Type 1 diabetes (T1DM) has been previously associated with northern latitude and vitamin D insufficiency. This study investigates the geospatial association between average daily ultraviolet B (UVB) irradiance and T1DM across the province of Newfoundland (NL), Canada. NL has one of the highest documented incidences of T1DM worldwide. A complete list of patients diagnosed (1987-2005) with T1DM in the province of Newfoundland and Labrador (NL) was constructed using multiple sources. All places of habitation at diagnosis were ascertained. Ecological analysis using Bayesian estimation was performed employing both NASA UVB data and latitude. Correlation of T1DM to both UVB irradiation and latitude was measured. A statistically significant correlation of erythemal UVB irradiance was observed (-0.0284: 95% CI -0.0542 to -0.0096). A more significant correlation of T1DM was observed with erythemal UVB irradiance than with latitude. This study suggests that erythemal UVB radiation may be geospatially associated with the incidence of T1DM in NL.

Observations on recent studies showing increased co-occurrence of autoimmune diseases.

The co-occurrence of autoimmune diseases has been epidemiologically studied and has aided in our understanding of autoimmunity. However, as new perspectives develop on the pathogenesis and natural history of autoimmune diseases, a refinement in the methodology for the study of the co-occurrence of disease is warranted in order to maximize the information that one may realize from such studies. This paper presents some recent results of co-occurrence studies and then proposes several refinements in the design of epidemiological studies in light of current understanding of the natural history of autoimmune diseases. It also suggests an historical perspective on the results of past studies as to the type of information that can be inferred from them.