ABSTRACT
OBJECTIVE: To compare the unilateral signs of knee osteoarthritis (KOA) 30 and 60 days after anterior cruciate ligament transection (ACLT). Pain, gait function, synovial fluid inflammation, and histopathological changes in the synovial membrane were analyzed, as well as the interaction between the variables. MATERIALS AND METHODS: Male Wistar rats (n = 32; 219.2 ± 18.6 g) were randomly distributed into four groups of eight animals each. Two groups were submitted to unilateral ACLT surgery to induce KOA and analyzed after 30 (KOA30) and 60 days (KOA60). Two control groups (without surgery) were also assessed after the same time periods (C30 and C60). All the groups were evaluated before ACLT from the least to most stressful tests (skin temperature, mechanical response threshold, gait test, thermal response threshold, and joint swelling), as well as 30 and 60 days after surgery. After euthanasia, the synovial fluid and synovial membrane were collected. RESULTS: Thirty days after ACLT, KOA30 showed decrease paw print area and mechanical response threshold, higher joint swelling, skin temperature, leukocyte count, cytokine levels, and synovitis score. No differences were found between KOA30 and KOA60. CONCLUSION: Our data showed that 30 days after ACLT is sufficient to induce signs of KOA in rats, such as pain, functional impairment, and synovial inflammation, suggesting that a shorter time period can be used as an experimental model.
Subject(s)
Anterior Cruciate Ligament/surgery , Inflammation/metabolism , Osteoarthritis, Knee/physiopathology , Animals , Cell Movement , Cytokines/metabolism , Disease Models, Animal , Knee Joint/pathology , Leukocytes/cytology , Male , Osteoarthritis, Knee/etiology , Pain Measurement , Rats , Rats, Wistar , Skin Temperature , Synovial Fluid/chemistry , Synovial Fluid/cytology , Synovial Membrane/pathologyABSTRACT
We report 2 fatal cases of congenital Zika virus (ZIKV) infection. Brain anomalies, including atrophy of the cerebral cortex and brainstem, and cerebellar aplasia were observed. The spinal cord showed architectural distortion, severe neuronal loss, and microcalcifications. The ZIKV proteins and flavivirus-like particles were detected in cytoplasm of spinal neurons, and spinal cord samples were positive for ZIKV RNA.
Subject(s)
Pregnancy Complications, Infectious , Spinal Cord Diseases , Spinal Cord/abnormalities , Zika Virus Infection , Zika Virus , Fatal Outcome , Female , Humans , Infant, Newborn , Male , Pregnancy , Pregnancy Complications, Infectious/pathology , Pregnancy Complications, Infectious/virology , Spinal Cord Diseases/congenital , Spinal Cord Diseases/pathology , Spinal Cord Diseases/virology , Zika Virus Infection/congenital , Zika Virus Infection/pathology , Zika Virus Infection/virologyABSTRACT
BACKGROUND: Steatosis is a risk factor in partial hepatectomy (PH) under ischaemia-reperfusion (I/R), which is commonly applied in clinical practice to reduce bleeding. Nutritional support strategies, as well as the role of peripheral adipose tissue as energy source for liver regeneration, remain poorly investigated. AIMS: To investigate whether the administration of either glucose or a lipid emulsion could protect steatotic and non-steatotic livers against damage and regenerative failure in an experimental model of PH under I/R. The relevance of peripheral adipose tissue in liver regeneration following surgery is studied. METHODS: Steatotic and non-steatotic rat livers were subjected to surgery and the effects of either glucose or lipid treatment on damage and regeneration, and part of the underlying mechanisms, were investigated. RESULTS: In non-steatotic livers, treatment with lipids or glucose provided the same protection against damage, regeneration failure and ATP drop. Adipose tissue was not required to regenerate non-steatotic livers. In the presence of hepatic steatosis, lipid treatment, but not glucose, protected against damage and regenerative failure by induction of cell cycle, maintenance of ATP levels and elevation of sphingosine-1-phosphate/ceramide ratio and phospholipid levels. Peripheral adipose tissue was required for regenerating the steatotic liver but it was not used as an energy source. CONCLUSION: Lipid treatment in non-steatotic livers provides the same protection as that afforded by glucose in conditions of PH under I/R, whereas the treatment with lipids is preferable to reduce the injurious effects of liver surgery in the presence of steatosis.
Subject(s)
Fatty Liver/metabolism , Glucose/pharmacology , Hepatectomy/adverse effects , Ischemia/metabolism , Lipids/pharmacology , Reperfusion , Adenosine Triphosphate/metabolism , Analysis of Variance , Animals , Ceramides , Fatty Liver/drug therapy , Fatty Liver/surgery , Glucose/metabolism , Ischemia/etiology , Liver/drug effects , Liver/physiology , Lysophospholipids , Rats , Regeneration/drug effects , Regeneration/physiology , Sphingosine/analogs & derivativesABSTRACT
In this study, the changes in oncogenic and tumor suppressor signaling pathways in liver and their association with serum and urinary biomarkers of aflatoxin exposure were evaluated in Wistar rats fed diets containing aflatoxin B1 (AFB1) for 90 days. Rats were divided into four groups (n = 15 per group) and assigned to dietary treatments containing 0 (control), 50 (AFB50), 100 (AFB100) and 200 µg AFB1 kg-1 diet (AFB200). Multiple preneoplastic foci of hepatocytes marked with glutathione-S-transferase-placental form (GST-P) were identified in AFB100 and AFB200 groups. Hepatocellular damage induced by AFB1 resulted in overexpression of cyclin D1 and ß-catenin. The liver expression of retinoblastoma (Rb) and p27Kip1 decreased in AFB100 and AFB200 groups, confirming the favorable conditions for neoplastic progression to hepatocellular carcinoma. All samples from rats fed AFB1-contaminated diets had quantifiable AFB1-lysine in serum or urinary AFM1 and AFB1-N7-guanine, with mean levels of 20.42-50.34 ng mL-1, 5.31-37.68 and 39.15-126.37 ng mg-1 creatinine, respectively. Positive correlations were found between AFB1-lysine, AFM1 or AFB1-N7-guanine and GST-P+, ß-catenin+ and cyclin D1+ hepatocytes, while Rb + cells negatively correlated with those AFB1 exposure biomarkers. The pathways evaluated are critical molecular mechanisms of AFB1-induced hepatocarcinogenesis in rats.
Subject(s)
Aflatoxin B1/toxicity , Cyclin D1/metabolism , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Retinoblastoma Protein/metabolism , beta Catenin/metabolism , Aflatoxin B1/analogs & derivatives , Aflatoxin B1/blood , Aflatoxin B1/metabolism , Aflatoxin B1/urine , Aflatoxin M1/urine , Alanine Transaminase/metabolism , Animals , Aspartate Aminotransferases/metabolism , Biomarkers/blood , Biomarkers/urine , Gene Expression/drug effects , Guanine/analogs & derivatives , Guanine/urine , Hepatocytes/drug effects , Liver/drug effects , Liver/pathology , Lysine/blood , Male , Precancerous Conditions/chemically induced , Precancerous Conditions/pathology , Rats, WistarABSTRACT
Th17 cell differentiation and pathogenicity depend on metabolic reprogramming inducing shifts toward glycolysis. Here, we show that the pyruvate kinase M2 (PKM2), a glycolytic enzyme required for cancer cell proliferation and tumor progression, is a key factor mediating Th17 cell differentiation and autoimmune inflammation. We found that PKM2 is highly expressed throughout the differentiation of Th17 cells in vitro and during experimental autoimmune encephalomyelitis (EAE) development. Strikingly, PKM2 is not required for the metabolic reprogramming and proliferative capacity of Th17 cells. However, T cell-specific PKM2 deletion impairs Th17 cell differentiation and ameliorates symptoms of EAE by decreasing Th17 cell-mediated inflammation and demyelination. Mechanistically, PKM2 translocates into the nucleus and interacts with STAT3, enhancing its activation and thereby increasing Th17 cell differentiation. Thus, PKM2 acts as a critical nonmetabolic regulator that fine-tunes Th17 cell differentiation and function in autoimmune-mediated inflammation.
Subject(s)
Autoimmunity/physiology , Inflammation/metabolism , Pyruvate Kinase/physiology , STAT3 Transcription Factor/metabolism , Th17 Cells/physiology , Animals , Cell Differentiation , Encephalomyelitis, Autoimmune, Experimental/metabolism , Encephalomyelitis, Autoimmune, Experimental/physiopathology , Flow Cytometry , Fluorescent Antibody Technique , Mice , Mice, Inbred C57BL , Pyruvate Kinase/metabolism , Real-Time Polymerase Chain Reaction , Th17 Cells/metabolismABSTRACT
We examined whether angiotensin (Ang) II receptor antagonists could be considered a therapeutic strategy in steatotic and nonsteatotic livers in conditions of partial hepatectomy under ischemia-reperfusion (I/R), which is commonly applied in clinical practice to reduce blood loss. We report that Ang II type I receptor (AT1R) antagonist, but not Ang II type II receptor (AT2R) antagonist, increased regeneration in nonsteatotic livers. In the presence of steatosis, both AT1R and AT2R antagonists increased liver regeneration. This effect was stronger when the two were combined. Neither of the Ang II receptor antagonists protected nonsteatotic livers against damage. Only the AT1R antagonist, through nitric oxide inhibition, reduced damage in steatotic livers. The combination of the AT1R and AT2R antagonists in steatotic livers conferred a similar degree of protection to AT1R antagonist alone. Herein, we show that p38 mitogen-activated protein kinase (p38) was a key mechanism in the regeneration induced by the Ang II receptor antagonists in both liver types because when this signaling pathway was inhibited, the beneficial effects of the Ang II receptor antagonists on liver regeneration disappeared, regardless of hepatocyte growth factor or transforming growth factor beta-hepatic levels. In conclusion, in conditions of partial hepatectomy under I/R, the AT1R antagonist for nonsteatotic livers and the AT1R and AT2R antagonists for steatotic livers improved regeneration in the remnant liver through p38 activation. In addition, the combination of the AT1R and AT2R antagonists in steatotic livers led to stronger liver regeneration than either antagonists used separately and also provided the same protection against damage as that afforded by AT1R antagonist alone.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/pharmacology , Angiotensin II Type 2 Receptor Blockers , Fatty Liver/drug therapy , Hepatectomy , Liver/pathology , Reperfusion Injury/drug therapy , Animals , Blotting, Western , Cell Proliferation/drug effects , Enzyme Inhibitors/pharmacology , Fatty Liver/pathology , Hepatocytes/drug effects , Nitric Oxide/metabolism , Nitric Oxide Synthase/antagonists & inhibitors , Obesity/drug therapy , Obesity/pathology , Rats , Rats, Zucker , Receptor, Angiotensin, Type 1/genetics , Receptor, Angiotensin, Type 2/genetics , Reperfusion Injury/pathology , Reverse Transcriptase Polymerase Chain Reaction , p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
Neutrophil influx is essential for corneal regeneration (Gan et al. 1999). KM+, a lectin from Artocarpus integrifolia, induces neutrophil migration (Santos-de-Oliveira et al. 1994). This study aims at investigating a possible effect of KM+ on corneal regeneration in rabbits. A 6.0-mm diameter area of debridement was created on the cornea of both eyes by mechanical scraping. The experimental eyes received drops of KM+ (2.5 microg/ml) every 2 h. The control eyes received buffer. The epithelial wounded areas of the lectin-treated and untreated eyes were stained with fluorescein, photographed and measured. The animals were killed 12 h (group 1, n = 5), 24 h (group 2, n = 10) and 48 h (group 3, n = 5) after the scraping. The corneas were analysed histologically (haematoxylin and eosin and immunostaining for proliferation cell nuclear antigen, p63, vascular endothelial growth factor, c-Met and laminin). No significant differences were found at the epithelial gap between treated and control eyes in the group 1. However, the number of neutrophils in the wounded area was significantly higher in treated eyes in this group. Three control and seven treated eyes were healed completely and only rare neutrophils persisted in the corneal stroma in group 2. No morphological distinction was observed between treated and control eyes in group 3. In treated corneas of group 2, there was an increase in immunostaining of factors involved in corneal healing compared to controls. Thus, topical application of KM+ may facilitate corneal epithelial wound healing in rabbits by means of a mechanism that involves increased influx of neutrophils into the wounded area induced by the lectin.
Subject(s)
Epithelium, Corneal/injuries , Mannose-Binding Lectins/pharmacology , Plant Lectins/pharmacology , Wound Healing/drug effects , Animals , Drug Evaluation, Preclinical/methods , Epithelium, Corneal/drug effects , Epithelium, Corneal/pathology , Male , Neutrophil Infiltration/drug effects , Rabbits , Wound Healing/physiologyABSTRACT
We have used an experimental model of aorta stenosis, with a Plexiglas plug, simulating a stable atheromatous plaque that promotes local turbulence and thrombosis. With animal survival of more than 24 h, we followed the partial fibrinolysis of the thrombus as well as its posterior organization and incorporation to the arterial wall as a neointima for up to 30 days. The mushroom plug form permitted the development of recirculation and stasis areas around it, favouring this evolution. Despite noted limitations, this study demonstrates that thrombus incorporation can contribute to plaque extension, as it can promote recirculation and stasis areas.
Subject(s)
Aortic Valve Stenosis/complications , Thrombosis/etiology , Animals , Aorta/ultrastructure , Aortic Valve Stenosis/diagnostic imaging , Aortic Valve Stenosis/pathology , Aortic Valve Stenosis/physiopathology , Disease Models, Animal , Hemorheology , Male , Microscopy, Electron , Microscopy, Electron, Scanning , Rats , Rats, Wistar , Thrombosis/pathology , Thrombosis/physiopathology , Tunica Intima/pathology , Tunica Media/pathology , Ultrasonography, Doppler, Color/methodsABSTRACT
BACKGROUND/AIMS: Nuclear factor kappa B (NFkappaB) plays important role in the pathogenesis of skeletal muscle ischemia/reperfusion (I/R) injury. Caffeic acid phenyl ester (CAPE), a potent NFkappaB inhibitor, exhibits protective effects on I/R injury in some tissues. In this report, the effect of CAPE on skeletal muscle I/R injury in rats was studied. METHODS: Wistar rats were submitted to sham operation, 120-min hindlimb ischemia, or 120-min hindlimb ischemia plus saline or CAPE treatment followed by 4-h reperfusion. Gastrocnemius muscle injury was evaluated by serum aminotransferase levels, muscle edema, tissue glutathione and malondialdehyde measurement, and scoring of histological damage. Apoptotic nuclei were determined by a terminal uridine deoxynucleotidyl transferase dUTP nick end labeling assay. Muscle neutrophil and mast cell accumulation were also assessed. Lipoperoxidation products and NFkappaB were evaluated by 4-hydroxynonenal and NFkappaB p65 immunohistochemistry, respectively. RESULTS: Animals submitted to ischemia showed a marked increase in aminotransferases after reperfusion, but with lower levels in the CAPE group. Tissue glutathione levels declined gradually during ischemia to reperfusion, and were partially recovered with CAPE treatment. The histological damage score, muscle edema percentage, tissue malondialdehyde content, apoptosis index, and neutrophil and mast cell infiltration, as well as 4-hydroxynonenal and NFkappaB p65 labeling, were higher in animals submitted to I/R compared with the ischemia group. However, the CAPE treatment significantly reduced all of these alterations. CONCLUSIONS: CAPE was able to protect skeletal muscle against I/R injury in rats. This effect may be associated with the inhibition of the NFkappaB signaling pathway and decrease of the tissue inflammatory response following skeletal muscle I/R.
Subject(s)
Caffeic Acids/therapeutic use , Muscle, Skeletal/injuries , NF-kappa B/antagonists & inhibitors , Phenylethyl Alcohol/analogs & derivatives , Reperfusion Injury/drug therapy , Animals , Caffeic Acids/pharmacology , Male , Medicine, Traditional , Phenylethyl Alcohol/pharmacology , Phenylethyl Alcohol/therapeutic use , Rats , Rats, WistarABSTRACT
BACKGROUND/AIMS: The transcription factor nuclear factor-kappa B (NF-kappaB) exerts a pivotal role in the pathogenesis of hepatic ischemia/reperfusion (I/R) injury. Caffeic acid phenyl ester (CAPE), a potent and specific NF-kappaB inhibitor, presents protective effects on I/R injury in some tissues. This study aimed to evaluate the effect of CAPE on hepatic I/R injury in rats. MATERIALS AND METHODS: Wistar rats were submitted to a sham operation, 60 min ischemia, or 60 min ischemia plus saline or CAPE treatment followed by 6 h reperfusion. Liver tissue injury was evaluated by alanine aminotransferase, aspartate aminotransferase, and tissue glutathione measurement, and histological damage score. Apoptotic hepatocytes were determined by the transferase-mediated dUTP-biotin nick-end labeling assay. Hepatic neutrophil accumulation was assessed by the naphthol method. Lipid peroxidation and NF-kappaB activation were evaluated by 4-hydroxynonenal and NF-kappaB p65 immunohistochemistry, respectively. RESULTS: Animals submitted to ischemia showed a marked increase of alanine aminotransferase and aspartate aminotransferase after reperfusion, but with lower levels in CAPE group. Tissue glutathione content declined gradually during ischemia to reperfusion and was partially recovered with CAPE treatment. The histological damage score, apoptosis index, and neutrophil infiltration, as well as 4-hydroxynonenal and NF-kappaB p65 nuclear labeling, were higher in the liver of animals submitted to I/R compared to the ischemia group. However, the CAPE treatment significantly reduced all of these alterations. CONCLUSIONS: CAPE was able to protect the liver against normothermic I/R injury in rats. This effect may be associated with the inhibition of the NF-kappaB signaling pathway and decrease of the acute inflammatory response following I/R in the liver.
Subject(s)
Caffeic Acids/therapeutic use , Liver/injuries , Phenylethyl Alcohol/analogs & derivatives , Reperfusion Injury/prevention & control , Alanine Transaminase/blood , Animals , Apoptosis , Aspartate Aminotransferases/blood , Glutathione/metabolism , Lipid Peroxidation , Liver/metabolism , Male , NF-kappa B/metabolism , Neutrophils/metabolism , Phenylethyl Alcohol/therapeutic use , Rats , Rats, WistarABSTRACT
In this study, hepatic biopsies from autopsy cases in São Paulo, Brazil, showing hepatocellular carcinoma (HCC, n = 8), cirrhosis associated with viral hepatitis (VC, n = 20), cirrhosis associated with alcoholism (AC, n = 20), and normal livers (NL or controls, n = 10) were subjected to determination of aflatoxin B1 (AFB1) and its main metabolites, and of markers of hepatic carcinogenesis Only non-metabolized AFB1 was detected in 13 samples (27.1%, N = 48) of liver disorders (HCC, VC and AC), at levels between 10.0 and 418.0 pg/g (mean: 76.6 ± 107.7 pg/g). Immuno-labeling of p53, cyclin D1, p21, ß-catenin, and Prohibitin (PB) increased mainly in HCC patients, in relation to the controls. AFB1+ samples of HCC presented higher expressions of p53, cyclin D1, p21, and ß-catenin compared with AFB1-livers. In contrast, p27, p16, and Rb immuno-labeling decreased in HCC, VC, and AC samples, compared with NL, with lowest values in AFB1+ samples for all liver disorders. Compared with NL, gene expression of cyclin D1 and PB in AFB1+ samples of HCC and AC were also higher, along with higher gene expression of p21 in VC and AC AFB1+ livers. Results indicated that patients with liver disorders were exposed to dietary aflatoxins, and that residual AFB1 in liver negatively affected the p53 and protein Rb pathways in HCC. Moreover, the presence of AFB1 in cirrhotic livers warrants concern about the potential contribution of dietary aflatoxin to disease progression during VC and AC.
ABSTRACT
Sepsis is an overwhelming systemic inflammation resulting from an uncontrolled infection that causes extensive tissue damage, organ dysfunction and eventually death. A growing body of evidence indicates that impaired neutrophil migration to the site of infection is associated with poor outcome in sepsis. Here we show that galectin-3 (Gal-3), an endogenous glycan-binding protein, plays a critical role in sepsis outcome. We found that serum Gal-3 concentration increased in patients with septic shock and mice undergoing sepsis induced by cecal ligation and puncture (CLP). Mice deficient in Gal-3 (Gal-3 KO) are more resistant to sepsis induced by CLP, showing lower levels of biochemical markers and neutrophil infiltration for organ injury/dysfunction than those observed in wild-type mice (WT). Furthermore, Gal-3 KO mice show an increased number of neutrophils in the primary focus of infection and reduced bacterial loads in the peritoneal cavity, blood, and lungs. Mechanistically, blood neutrophils from septic mice show higher levels of surface-bound Gal-3 than neutrophils from naive mice. The deficiency of Gal-3 was associated with increased rolling and adhesion of these cells in mesenteric venules. Our results indicate that Gal-3, secreted during sepsis, inhibits neutrophil migration into the infectious focus, which promotes the bacterial spread and worsens the outcome of sepsis.
Subject(s)
Coinfection/blood , Coinfection/immunology , Galectin 3/blood , Neutrophil Infiltration , Sepsis/immunology , Sepsis/microbiology , Aged , Animals , Blood Proteins , Disease Models, Animal , Female , Galectin 3/immunology , Galectins , Humans , Lung/microbiology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Middle Aged , Peritoneum/microbiologyABSTRACT
Sepsis is a systemic inflammatory response as a result of uncontrolled infections. Neutrophils are the first cells to reach the primary sites of infection, and chemokines play a key role in recruiting neutrophils. However, in sepsis chemokines could also contribute to neutrophil infiltration to vital organs leading to multiple organ failure. ACKR2 is an atypical chemokine receptor, which can remove and degrade inflammatory CC chemokines. The role of ACK2 in sepsis is unknown. Using a model of cecal ligation and puncture (CLP), we demonstrate here that ACKR2 deficient () mice exhibited a significant reduction in the survival rate compared with similarly treated wild-type (WT) mice. However, neutrophil migration to the peritoneal cavity and bacterial load were similar between WT and ACKR2 mice during CLP. In contrast, ACKR2 mice showed increased neutrophil infiltration and elevated CC chemokine levels in the lung, kidney, and heart compared with the WT mice. In addition, ACKR2 mice also showed more severe lesions in the lung and kidney than those in the WT mice. Consistent with these results, WT mice under nonsevere sepsis (90% survival) had higher expression of ACKR2 in these organs than mice under severe sepsis (no survival). Finally, the lungs from septic patients showed increased number of ACKR2 cells compared with those of nonseptic patients. Our data indicate that ACKR2 may have a protective role during sepsis, and the absence of ACKR2 leads to exacerbated chemokine accumulation, neutrophil infiltration, and damage to vital organs.
Subject(s)
Multiple Organ Failure/metabolism , Neutrophil Infiltration , Neutrophils/metabolism , Receptors, Chemokine/metabolism , Sepsis/metabolism , Animals , Disease Models, Animal , Female , Male , Mice , Multiple Organ Failure/pathology , Neutrophils/pathology , Sepsis/pathologyABSTRACT
BACKGROUND: The inhibition of pyrimidine biosynthesis by blocking the dihydroorotate dehydrogenase (DHODH) activity, the prime target of leflunomide (LEF), has been proven to be an effective strategy for rheumatoid arthritis (RA) treatment. However, a considerable proportion of RA patients are refractory to LEF. Here, we investigated lapachol (LAP), a natural naphthoquinone, as a potential DHODH inhibitor and addressed its immunosuppressive properties. METHODS: Molecular flexible docking studies and bioactivity assays were performed to determine the ability of LAP to interact and inhibit DHODH. In vitro studies were conducted to assess the antiproliferative effect of LAP using isolated lymphocytes. Finally, collagen-induced arthritis (CIA) and antigen-induced arthritis (AIA) models were employed to address the anti-arthritic effects of LAP. RESULTS: We found that LAP is a potent DHODH inhibitor which had a remarkable ability to inhibit both human and murine lymphocyte proliferation in vitro. Importantly, uridine supplementation abrogated the antiproliferative effect of LAP, supporting that the pyrimidine metabolic pathway is the target of LAP. In vivo, LAP treatment markedly reduced CIA and AIA progression as evidenced by the reduction in clinical score, articular tissue damage, and inflammation. CONCLUSIONS: Our findings propose a binding model of interaction and support the ability of LAP to inhibit DHODH, decreasing lymphocyte proliferation and attenuating the severity of experimental autoimmune arthritis. Therefore, LAP could be considered as a potential immunosuppressive lead candidate with potential therapeutic implications for RA.
Subject(s)
Arthritis, Experimental/drug therapy , Arthritis, Rheumatoid/drug therapy , Immunosuppressive Agents/pharmacology , Naphthoquinones/pharmacology , Animals , CD4-Positive T-Lymphocytes/drug effects , Cell Proliferation/drug effects , Dihydroorotate Dehydrogenase , Humans , Lymphocyte Activation/drug effects , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Molecular Docking Simulation , Oxidoreductases Acting on CH-CH Group Donors/antagonists & inhibitors , Rats , Rats, WistarABSTRACT
PURPOSE: Strabismus is an oculomotor disorder in which there is a misalignment of the visual axes of the eyes. Inferior oblique muscle (IOM) overaction is a common finding in comitant horizontal strabismus, but its origin is unclear. Recent studies have demonstrated that myogenic satellite cells (SCs) are still activated in adult extraocular muscles, with continuous myonuclear addition in normal uninjured muscles. The objective of this study was to determine whether there are differences in the processes of activation and proliferation of SCs in IOMs of patients with strabismus and IOM overaction and in patients with no history of strabismus. METHODS: Cross sections of IOMs from strabismic and control groups were analyzed immunohistochemically for the presence of MyoD1 and myogenin, specific markers of activated SCs, and for c-Met, which is expressed in quiescent, activated, and proliferating SCs. RESULTS: In overacting IOMs of 26 patients in the strabismic group and 10 patients in the control group, 28.8% and 3.0% of the myofibers, respectively, were associated with MyoD1-positive SC. The frequency of myogenin-positive SC was 30.8% in the strabismic group and 3.6% in the control group, and the frequency of presumptive SCs immunostained for c-Met was 33.6% in the strabismic group and 34.1% in the control group. CONCLUSIONS: The presence of an increased number of activated SCs in overacting IOMs of the strabismic group in contrast to the frequency in the control group resembles the findings detected in developing, regenerating, or hypertrophic muscle tissue. High levels of MyoD1- and myogenin-positive SC in overacting IOMs support the hypothesis that these cells may be involved in alterations in IOM structure correlated with the overaction observed clinically.
Subject(s)
Oculomotor Muscles/pathology , Satellite Cells, Skeletal Muscle/pathology , Strabismus/pathology , Adolescent , Adult , Cell Proliferation , Child , Child, Preschool , Female , Humans , Immunoenzyme Techniques , Male , Middle Aged , MyoD Protein/metabolism , Myogenin/metabolism , Oculomotor Muscles/metabolism , Satellite Cells, Skeletal Muscle/metabolism , Strabismus/metabolismABSTRACT
This study compares the populations of liver mesenchymal cells (LMCs) and their proliferative activity in schistosomal periportal fibrosis and in hepatitis C virus-induced cirrhosis. LMCs were evaluated by immunohistochemical double staining for proliferating cell nuclear antigen (PCNA) and alpha-smooth muscle actin (alpha-SMA) or glial fibrillary acid protein (GFAP) in liver biopsies from humans with schistosomal fibrosis (n=40), hepatitis C virus-induced cirrhosis (n=20), and normal controls (n=20). The number of LMCs was found to be higher in schistosomal fibrosis than in the normal liver, but lower than in cirrhosis. alpha-SMA- and GFAP-positive cells were increased in both diseases, but more so in cirrhosis. In cirrhotic liver, alpha-SMA-positive cells were highly predominant in relation to GFAP-positive cells. However, there was an inverted ratio between these cells in schistosomiasis as compared to cirrhosis. The PCNA labeling index was higher in alpha-SMA-positive cells than in GFAP-positive cells, and did not differ between pipe-stem fibrosis and liver cirrhosis regarding alpha-SMA- or GFAP-positive cells. The predominance of GFAP-positive cells observed in schistosomiasis suggests that hepatic stellate cells (HSCs) have a major role in connective tissue deposition in the human schistosomal liver. On the other hand, the smaller number of LMCs in schistosomal fibrosis in comparison to liver cirrhosis may be related to mild and limited injury due to the schistosomal egg-induced inflammatory response. The granulomatous inflammation around Schistosoma mansoni eggs appears to mobilize and activate a reduced number of mesenchymal cells in comparison to the scattered necro-inflammatory reaction produced by the hepatitis C virus.
Subject(s)
Hepatocytes/pathology , Liver Cirrhosis/pathology , Schistosomiasis mansoni/pathology , Actins/metabolism , Adult , Female , Glial Fibrillary Acidic Protein/metabolism , Hepatocytes/metabolism , Humans , Immunohistochemistry , Liver Cirrhosis/metabolism , Male , Middle Aged , Schistosomiasis mansoni/metabolismABSTRACT
ArtinM is a d-mannose-binding lectin found in the seeds of Artocarpus heterophyllus (jackfruit) that interacts with N-glycans, that is associated with receptors on the surface of phagocytic cells and induces the production of inflammatory mediators. Some of them are especially important because they may be required for antitumor immune response. This study aimed to evaluate the effect of ArtinM on hepatocellular preneoplastic foci. Wistar rats received 50 mg/kg of diethyl-nitrosamine (DEN) intraperitoneal weekly for 12 weeks. From the 14th week, the treated animals received 50 µg/kg of ArtinM subcutaneous every 2 weeks until the 18th week, whereas control animals were injected with vehicle alone. Preneoplastic-related factors were estimated using histological, western blotting and RT-PCR analysis. In comparison to the groups exposed to DEN, the ArtinM-treated rats showed diminution of preneoplastic foci, decreased expression of proliferating cell nuclear antigen (PCNA), increased number of nuclear p21 and p27 stained cells, augmented number of apoptotic cells, increased expression of p53, p42/44 MAPK and p21 proteins, reduced cyclin D1 (CCND1) protein levels and increased expression of TNFα and IFNγ genes. No difference was observed in interleukin 12 (IL12) protein levels. These findings indicate that ArtinM may provide protection against hepatocarcinogenesis as a result of the induction of cell-cycle blockage and pro-apoptotic mechanisms.
Subject(s)
Apoptosis/drug effects , Cell Proliferation/drug effects , Liver Neoplasms/prevention & control , Mannose-Binding Lectin/pharmacology , Precancerous Conditions/prevention & control , Animals , Apoptosis/genetics , Apoptosis Regulatory Proteins/metabolism , Artocarpus/chemistry , Blotting, Western , Cell Proliferation/genetics , Cyclin D1/metabolism , Gene Expression Regulation, Neoplastic/drug effects , Interferon-gamma/genetics , Interleukin-12/genetics , Liver Neoplasms/genetics , Liver Neoplasms/metabolism , Male , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Phytotherapy , Precancerous Conditions/genetics , Precancerous Conditions/metabolism , Proliferating Cell Nuclear Antigen/metabolism , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Tumor Necrosis Factor-alpha/geneticsABSTRACT
OBJECTIVES: To evaluate the discrepancies between clinical and autopsy diagnoses in patients who died in the pediatric intensive care units (PICUs) of a tertiary care university hospital. METHODS: A prospective study of all consecutive autopsies discussed at monthly mortality conferences over 5 years. Discrepancies between premortem and autopsy diagnoses were classified according to modified Goldman et al criteria. RESULTS: From January 1, 2011, to December 31, 2015, a total of 2,679 children were admitted to the two PICUs of our hospital; 257 (9.6%) died, 150 (58.4%) underwent autopsy, and 123 were included. Complete concordance between clinical and postmortem diagnoses was observed in 86 (69.9%) patients; 20 (16.3%) had a class I discrepancy, and eight (6.5%) had a class II discrepancy. Comparing 2011 and 2015, the rate of major discrepancies decreased from 31.6% to 15%. CONCLUSIONS: Our results emphasize the importance of autopsy to clarify the cause of death and its potential contribution to improvement of team performance and quality of care.
Subject(s)
Autopsy , Cause of Death , Critical Illness , Adolescent , Child , Child, Preschool , Diagnostic Errors , Female , Humans , Infant , Infant, Newborn , Intensive Care Units , Male , Prospective StudiesABSTRACT
BACKGROUND: Bradykinin (BK) is used in different tissues. Dose-dependent studies have demonstrated that low doses protect against ischemia/reperfusion (I/R) injury while higher doses lead to adverse effects. Although the beneficial effects of BK infusion were observed in myocardium, its role on the I/R impact in skeletal muscle (SM) has not been fully clarified. OBJECTIVE: This study was carried out to evaluate the effects of BK, administered in the hindlimbs of rats subjected to I/R. METHODS: The study design included three experimental groups: Group 1 control (saline), Group 2 (bradykinin), and Group 3 (HOE 140, a BK2 receptor blocker). In all three groups, rats were subjected to hindlimb ischemia for a total of 2 h followed by continuous 4 h of reperfusion with pharmacological interventions. The methods include analysis of enzymes (lactate dehydrogenase-LDH and creatinine phosphokinase-CPK), cell membrane marker of injury (malondialdeyde-MDA), recruitment of neutrophils (myeloperoxidase-MPO), and apoptosis index (immunohistochemistry TUNEL in situ peroxidase dead end). RESULTS: Except for the apoptotic index, all parameters studied were shown to be elevated in the reperfusion group intervened with BK. The blocking of BK2 receptors by HOE 140 did not affect the I/R injury. CONCLUSION: After 2 h of total ischemia, infusion of bradykinin during 4 h of reperfusion, worsened the I/R injury in the hindlimb skeletal muscle.
Subject(s)
Bradykinin B2 Receptor Antagonists/administration & dosage , Bradykinin/analogs & derivatives , Reperfusion Injury/prevention & control , Vasodilator Agents/administration & dosage , Animals , Apoptosis , Bradykinin/administration & dosage , Creatine Kinase/blood , Creatine Kinase/metabolism , Hindlimb/physiopathology , L-Lactate Dehydrogenase/metabolism , Male , Malondialdehyde/blood , Malondialdehyde/metabolism , Muscle, Skeletal/metabolism , Muscle, Skeletal/physiopathology , Peroxidase/blood , Peroxidase/metabolism , Rats , Rats, Wistar , Reperfusion Injury/blood , Reperfusion Injury/metabolismABSTRACT
BACKGROUND: Overexpression of the oncogene yes-associated-protein-1 (YAP1) is associated with increased cell proliferation in human cancers. YAP1 is a potential target of the Wnt/beta-catenin pathway, which plays an important role in adrenocortical tumors (ACT). The role of YAP1 in adrenocortical tumorigenesis has not been assessed. AIMS: To evaluate YAP1 expression in normal adrenals and pediatric ACT and its association with disease outcome. To investigate the interaction between YAP1 and the Wnt/beta-catenin pathway in adrenocortical cells. RESULTS: Strong YAP1 staining was present in fetal adrenals and pediatric ACT but weak in postnatal adrenals. In pediatric ACT, YAP1 mRNA overexpression was associated with death, recurrent/metastatic disease and lower overall survival. The inhibition of the Wnt/beta-catenin pathway increased YAP1 mRNA expression. siYAP1 increased CTNNB1/beta-catenin expression and nuclear staining regardless of DLV2, moreover, it decreased cell growth and impaired cell migration. MATERIALS AND METHODS: We assessed in 42 pediatric ACT samples the YAP1 protein expression by immunohistochemistry and mRNA expression by RT-qPCR and analyzed their association with outcome. As controls, we resort 32 fetal and postnatal normal adrenals for IHC and 10 normal adrenal cortices for RT-qPCR. The interaction between YAP1 and the Wnt/beta-catenin pathway was assessed in NCI-H295 adrenocortical cells by inhibiting the TCF/beta-catenin complex and by knocking down YAP1. CONCLUSION: YAP1 overexpression is a marker of poor prognosis for pediatric patients with ACT. In adrenocortical cells, there is a close crosstalk between YAP1 and Wnt/beta-catenin. These data open the possibility of future molecular therapies targeting Hippo/YAP1 signaling to treat advanced ACT.