Latent Murine Cytomegalovirus Infection Contributes to EAE Pathogenesis / Latentna Infekcija Mišjim Citomegalovirusom Ima Ulogu U Patogenezi Eksperimentalnog Autoimunskog Encefalomijelitisa

Virusna infekcija se navodi kao najverovatniji faktor okoline koji utiče na razvoj multiple skleroze (MS). Postoje konfliktni podaci o ulozi infekcije citomegalovirusom (CMV) u patogenezi multiple skleroze. Koristili smo BALB/c miševe, rezistentne na indukciju eksperimentalnog autoimunskog encefalomijelitisa (EAE), i mišji citomegalovirus (MCMV), mišji homolog humanom citomegalovirusu da ispitamo kako virusna infekcija može da utiče na razvoj autoimunske neuroinflamacije. Miševi sa latentnom neonatalnom infekcijom mišjim citomegalovirusom su razvili tipičan EAE. Slično kao u MS, MCMV EAE miševi su razvili infiltrate u centralnom nervnom sistemu (CNS) sa sličnom zastupljenošću CD4+ i CD8+ T limfocita. Uočen je influks i Th 1 i Th 17 ćelija u CNS MCMV EAE miševa. Interesantno je da razvoj autoimunske inflamacije nakon latentne MCMV infekcije prati značajan influks samo Tc17 (CD8+IL-17+ i CD8+RoRγt+), a ne i Tc1 ćelija. Naši rezultati ukazuju da latentna MCMV infekcija verovatno utiče na razvoj inflamatornih limfocita koji mogu da indukuju autoimunski proces u CNS-u, direktno pojačava razvoj patoloških procesa u CNS-u nakon indukcije EAE i ukazuje na CMV kao na mogući faktor okoline koji utiče na razvoj multiple skleroze i drugih autoimunskih bolesti

Molecular Aspects of MAFLD—New Insights on Pathogenesis and Treatment

Metabolic-associated liver disease (MAFLD) affects up to 70% of overweight and more than 90% of morbidly obese people, and its pathogenesis is rather complex and multifactorial. The criteria for MAFLD include the presence of hepatic steatosis in addition to one of the following three criteria: overweight or obesity, presence of type 2 diabetes mellitus (T2DM), or evidence of metabolic dysregulation. If the specific criteria are present, the diagnosis of MAFLD can be made regardless of alcohol consumption and previous liver disease. The pathophysiological mechanisms of MAFLD, including inflammation, lipotoxicity, mitochondrial disfunction, and oxidative stress, as well as the impact of intestinal gut microbiota, are constantly being elucidated. Treatment strategies that are continually emerging are based on different key points in MAFLD pathogenesis. Yet, the ideal therapeutic option has still not been found and future research is of great importance, as MAFLD represents a multisystemic disease with numerous complications

A Passively Suspended Tesla Pump Left Ventricular Assist Device

Detail of hoof and texture of the hair; Both wild boars (a male and female) were created in 1926 and cast in 1929. Carl Milles created them on order from an English lord, Lord Melchett. There are multiple casts, and one set was eventually acquired by the Swedish Royal Court and can now be found at Ulriksdal Palace. Carl Milles sculpted many animals. He was often interested in heavyset, large animals, such as bears, elephants and wild boars. There is a beetle and a lizard between the front legs of the boar. The Cranbrook casts stand guard over the painted iron Nichols Gate, designed by Eliel Saarinen. Source: Millesgården [estate website]; http://www.millesgarden.se/ (accessed 7/22/2015

The New Therapeutic Approaches in the Treatment of Non-Alcoholic Fatty Liver Disease

Non-alcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease which is characterized by extremely complex pathogenetic mechanisms and multifactorial etiology. Some of the many pathophysiological mechanisms involved in the development of NAFLD include oxidative stress, impaired mitochondrial metabolism, inflammation, gut microbiota, and interaction between the brain-liver-axis and the regulation of hepatic lipid metabolism. The new therapeutic approaches in the treatment of NAFLD are targeting some of these milestones along the pathophysiological pathway and include drugs like agonists of peroxisome proliferator-activated receptors (PPARs), glucagon-like peptide-1 (GLP-1) agonists, sodium/glucose transport protein 2 (SGLT2) inhibitors, farnesoid X receptor (FXR) agonists, probiotics, and symbiotics. Further efforts in biomedical sciences should focus on the investigation of the relationship between the microbiome, liver metabolism, and response to inflammation, systemic consequences of metabolic syndrome

Identification of the cell type that expresses IL-33 in inflammatory foci in infected liver tissue as F4/80⁺ macrophages.

<p>BALB/c mice were injected i.v. with 5x10⁵ PFU of WT MCMV (MW97.01) and liver tissue was harvested on day 5 p.i. (<b>A</b>) Consecutive serial 1-μm sections of liver tissue focusing on an infected hepatocyte (Hc) that is delimited from uninfected tissue by a sheath made up by a mononuclear cell infiltrate. The expression of the indicated marker molecules was tested in a two-color IHC (2C-IHC) staining. (<b>a</b>-<b>d</b>) Identification of the infected Hc by red staining of the intranuclear viral IE1 protein. (<b>a</b>) Focus-forming mononuclear cells are not CD31^<b>+</b> black-stained endothelial cells (EC). (<b>b</b>) Focus-forming mononuclear cells are not CD3ε^<b>+</b> black-stained cells, thus excluding α/ß and γ/δ T cells as well as NKT cells. (<b>c</b>) Identification of focus-forming mononuclear cells as black-stained F4/80^<b>+</b> macrophages (Mø). (<b>d</b>) IL33-expressing cells stained in turquoise-green color colocalize with focus-forming F4/80 macrophages in the neighboring section of image <b>c</b>. Counterstaining with hematoxylin. Arrows point to the indicated cell types exemplarily. The bar markers represent 50 μm throughout. (<b>B</b>) 2C-IHC verifying colocalization of F4/80 and IL33 on the cellular level. (<b>a</b>) Higher resolution image of an advanced, aged focus consisting of a cluster of dual-stained F4/80⁺ (red) IL33⁺ (turquoise-green) macrophages (Mø) surrounding an infected hepatocyte (Hc) that is identified by an intranuclear inclusion body. Note that dually-expressing macrophages localize also to liver tissue outside of a focus. (<b>b</b>) A young focus in which dual-stained F4/80^<b>+</b> (red) IL33⁺ (turquoise-green) macrophages cling to an infected hepatocyte (Hc) that shows the pathocytomorphology of an owl’s eye cell with an intranuclear inclusion body that indicates the late phase (L phase) in the viral gene expression program. Counterstaining with hematoxylin. Arrows point to sites of interest. The bar markers represent 50 μm.</p

Treg cells show an activated phenotype after MCMV infection.

<p>BALB/c mice were i.v. injected with 2x10⁵ PFU of WT MCMV (clone MW97.01) or left uninfected. (<b>A</b>) Absolute number of Treg cells in spleen and liver is shown. (<b>B</b>) Representative FACS plots and (<b>C</b>) graphs showing percentages and (<b>D</b>) median fluorescence intensity (MFI) of Ki-67 expression by naive Treg cells. (<b>F</b>) Bcl-2 expression by naive Treg cells. (<b>E</b>) Mice were treated with BrdU in drinking water for 6 days starting at the day of infection. Percentage of BrdU positive Treg cells on day 7 was determined. (<b>G</b>) Histograms show a representative expression of different markers by Treg cells from uninfected and 7 days infected mice. (<b>H</b>) Representative FACS plots and (<b>I</b>) graphs showing percentages and (<b>J</b>) median fluorescence intensity (MFI) of ST2 expression by Treg cells isolated from the spleen and liver of naive BALB/c and ST2^-/- mice. Data are shown as mean ± SEM of n = 3–5 mice from one representative experiment out of three. *p<0.05; ** p<0.01; ***p<0.001 from two tailed, unpaired Student’s t-test.</p

ST2^-/- mice show increased liver damage and mortality rate after MCMV infection.

<p>(<b>A</b>) BALB/c and ST2^-/- mice were i.v. injected with 2x10⁵ PFU of WT MCMV (MW97.01) and lymphocytes from spleen and liver were analyzed on day 7 p.i. Absolute number of Treg cells is shown. (<b>B</b>-<b>E</b>) BALB/c and ST2^-/- mice were i.v. injected with 10⁶ PFU of WT MCMV (pSM3fr-MCK-2fl clone 3.3) and analyzed on day 5 p.i. (<b>B</b>) AST and ALT serum levels were determined. (<b>C</b>) Scores of cumulative liver pathology for apoptosis, intranuclear inclusion bodies (INIBs), inflammation, and necrosis. Bars correspond to the mean score for each parameter. The height of each bar represents the mean of the total histological score (out of 12). (<b>D</b>) Representative H&E and (<b>E</b>) Caspase-3 staining of paraffin embedded liver sections. (<b>F</b>) BALB/c and ST2^-/- mice were i.p. injected with indicated doses of SGV MCMV. Survival rates were monitored daily. Data are shown as mean ± SEM of n = 3–5 mice from one representative experiment out of three. For survival monitoring n = 7. *p <0.05 and **p<0.01 from two tailed, unpaired Student’s t-test.</p

Intrinsic requirement for ST2 expression in Treg cells.

<p>Mixed bone marrow chimeras were generated by irradiation of C57BL/6 CD45.1⁺CD45.2⁺ mice followed by i.v. injection of 1:1 mixture of wild-type (WT; CD45.1⁺) and knockout (ST2^-/-; CD45.2⁺) bone marrow cells. After reconstitution, mixed chimeras were infected with 2x10⁵ PFU of Δm157 MCMV and analyzed on day 7 p.i. (<b>A</b>) Percentage of donor WT and KO Treg cells is shown. (<b>B</b>) The ratio between WT and KO Treg cells is shown. Dotted line represents initial ratio of transferred bone marrow cells. (<b>C</b>) Percentage of Ki-67 expression by donor Treg cells is shown. Data are shown as mean ± SEM of n = 4–5 mice per group from one representative experiment out of two. *p <0.05 and **p<0.01 from two tailed, unpaired Student’s t-test.</p