Ferritin accumulation in dystrophic microglia is an early event in the development of Huntington's disease

Huntington's Disease (HD) is characterized primarily by neuropathological changes in the striatum, including loss of medium-spiny neurons, nuclear inclusions of the huntingtin protein, gliosis, and abnormally high iron levels. Information about how these conditions interact, or about the temporal order in which they appear, is lacking. This study investigated if, and when, iron-related changes occur in the R6/2 transgenic mouse model of HD and compared the results with those from HD patients. Relative to wild-type mice, R6/2 mice had increased immunostaining for ferritin, an iron storage protein, in the striatum beginning at 2-4 weeks postnatal and in cortex and hippocampus starting at 5-7 weeks. The ferritin staining was found primarily in microglia, and became more pronounced as the mice matured. Ferritin-labeled microglia in R6/2 mice appeared dystrophic in that they had thick, twisted processes with cytoplasmic breaks; some of these cells also contained the mutant huntingtin protein. Brains from HD patients (Vonsattel grades 0-4) also had increased numbers of ferritin-containing microglia, some of which were dystrophic. The cells were positive for Perl's stain, indicating that they contained abnormally high levels of iron. These results provide the first evidence that perturbations to iron metabolism in HD are predominately associated with microglia and occur early enough to be important contributors to HD progression.     

Improving education in primary care: development of an online curriculum using the blended learning model

Background  

Standardizing the experiences of medical students in a community preceptorship where clinical sites vary by geography and discipline can be challenging. Computer-assisted learning is prevalent in medical education and can help standardize experiences, but often is not used to its fullest advantage. A blended learning curriculum combining web-based modules with face-to-face learning can ensure students obtain core curricular principles.     

Protein protein interaction inhibition (2P2I) combining high throughput and virtual screening: Application to the HIV-1 Nef protein

Protein-protein recognition is the cornerstone of multiple cellular and pathological functions. Therefore, protein-protein interaction inhibition (2P2I) is endowed with great therapeutic potential despite the initial belief that 2P2I was refractory to small-molecule intervention. Improved knowledge of complex molecular binding surfaces has recently stimulated renewed interest for 2P2I, especially after identification of "hot spots" and first inhibitory compounds. However, the combination of target complexity and lack of starting compound has thwarted experimental results and created intellectual barriers. Here we combined virtual and experimental screening when no previously known inhibitors can be used as starting point in a structure-based research program that targets an SH3 binding surface of the HIV type I Nef protein. High-throughput docking and application of a pharmacophoric filter on one hand and search for analogy on the other hand identified drug-like compounds that were further confirmed to bind Nef in the micromolar range (isothermal titration calorimetry), to target the Nef SH3 binding surface (NMR experiments), and to efficiently compete for Nef-SH3 interactions (cell-based assay, GST pull-down). Initial identification of these compounds by virtual screening was validated by screening of the very same library of compounds in the cell-based assay, demonstrating that a significant enrichment factor was attained by the in silico screening. To our knowledge, our results identify the first set of drug-like compounds that functionally target the HIV-1 Nef SH3 binding surface and provide the basis for a powerful discovery process that should help to speed up 2P2I strategies and open avenues for new class of antiviral molecules.     

Association between serum vitamin B12 levels and metabolic syndrome in a euthyroid population

Aims

To determine the association between serum levels of vitamin B12 and metabolic syndrome (MetS) in a population of euthyroid adults.

High free triiodothyronine and free-triiodothyronine-to-free-thyroxine ratio levels are associated with metabolic syndrome in a euthyroid population

Aims

To determine the association between free triiodothyronine (FT3), free thyroxine (FT4) and free-triiodothyronine-to-free-thyroxine ratio (FT3/FT4) levels and Metabolic Syndrome (MetS).

High triglycerides to HDL-cholesterol ratio is associated with insulin resistance in normal-weight healthy adults

AimTo evaluate the association between high triglyceride/HDL-cholesterol (TG/HDL-C) ratio and insulin resistance (IR) or hyperinsulinemia after oral glucose tolerance test (OGTT) in normal-weight healthy adults.MethodsWe carried out an analytical cross-sectional study in euthyroid non-diabetic adults, who attended the outpatient service of a private clinic in Lima-Peru from 2012 to 2016. Participants were divided in two groups according to the presence or absence of high TG/HDL-C ratio, IR or hyperinsulinemia after OGTT. TG/HDL-C ratio values ≥ 3 were considered as high. IR was defined as a Homeostasis Model Assessment (HOMA-IR) value ≥ 2.28 and hyperinsulinemia after OGTT as a serum insulin value ≥ 80μU/mL after 120 min of 75-g glucose intake. We elaborated crude and adjusted Poisson generalized linear models to evaluate the association between high TG/HDL-C ratio and IR or hyperinsulinemia after OGTT and reported the prevalence ratio (PR) with their respective 95% confidence intervals (95%CI).ResultsWe analyzed the data of 118 individuals. Prevalence of high TG/HDL-C ratio was 17.8% (n = 21) while the prevalence of IR and hyperinsulinemia after OGTT was 24.6% (n = 29) and 17.0% (n = 20), respectively. TG/HDL-C-ratio values were positively correlated with HOMA-IR (r = 0.498; p < 0.01) and serum insulin after OGTT (r = 0.326; p < 0.001). In the adjusted model, high TG/HDL-C ratio was associated with both IR (aPR = 3.16; 95%CI: 1.80–5.77) and hyperinsulinemia after OGTT (aPR = 2.36; 95%CI: 1.20–4.63).ConclusionsHigh TG/HDL-C ratio was associated with both IR markers used in our study, appearing to be a clinically useful tool to assess IR in euthyroid normal-weight adults without type 2 diabetes mellitus.     

In silico molecular target prediction unveils mebendazole as a potent MAPK14 inhibitor

The concept of polypharmacology involves the interaction of drug molecules with multiple molecular targets. It provides a unique opportunity for the repurposing of already‐approved drugs to target key factors involved in human diseases. Herein, we used an in silico target prediction algorithm to investigate the mechanism of action of mebendazole, an antihelminthic drug, currently repurposed in the treatment of brain tumors. First, we confirmed that mebendazole decreased the viability of glioblastoma cells in vitro (IC₅₀ values ranging from 288 nm to 2.1 µm). Our in silico approach unveiled 21 putative molecular targets for mebendazole, including 12 proteins significantly upregulated at the gene level in glioblastoma as compared to normal brain tissue (fold change > 1.5; P < 0.0001). Validation experiments were performed on three major kinases involved in cancer biology: ABL1, MAPK1/ERK2, and MAPK14/p38α. Mebendazole could inhibit the activity of these kinases in vitro in a dose‐dependent manner, with a high potency against MAPK14 (IC₅₀ = 104 ± 46 nm). Its direct binding to MAPK14 was further validated in vitro, and inhibition of MAPK14 kinase activity was confirmed in live glioblastoma cells. Consistent with biophysical data, molecular modeling suggested that mebendazole was able to bind to the catalytic site of MAPK14. Finally, gene silencing demonstrated that MAPK14 is involved in glioblastoma tumor spheroid growth and response to mebendazole treatment. This study thus highlighted the role of MAPK14 in the anticancer mechanism of action of mebendazole and provides further rationale for the pharmacological targeting of MAPK14 in brain tumors. It also opens new avenues for the development of novel MAPK14/p38α inhibitors to treat human diseases.

Abbreviations

BRET

bioluminescence resonance energy transfer

GBM

glioblastoma

GTeX

Genotype‐Tissue Expression

IC₅₀

half‐maximal inhibitory concentration

ITC

isothermal titration calorimetry

MBZ

mebendazole

nanoDSF

nanoscale differential scanning fluorimetry

qRT‐PCR

quantitative real‐time polymerase chain reaction

RT

room temperature

siRNA

small interfering RNA

TCGA

The Cancer Genome Atlas

TSA

thermal shift assay

     

Association between free thyroid hormones values and the lipid profile in middle-aged women with chronic symptoms

Aims

To determine the association between the thyroid hormones(FT3, FT4 and TSH) and the lipid profile markers(HDL-c, LDL-c and triglycerides) values in middle-aged women with no metabolic disorders and recurrent chronic symptomatology.