The Role of CD 34 Hematopoietic Progenitor Cells, Macrophages, and Smooth Muscle Cells in Human Coronary Artery Atherogenesis

BACKGROUND: Atherosclerosis is a widespread and devastating disease and one of the leading causes of death worldwide. So much is there to understand about atherosclerosis. And although a lot is already discovered, yet most of the studies are performed in cell cultures and animal models. Recent technologies for genetic engineering and imaging are mainly performed on animal models, with few studies in human tissues. A better understanding of their role is required. AIM: We aim to study the expression of CD 34 hematopoietic progenitor stem cell, CD 68 macrophages, and smooth muscle actin (SMA)-positive smooth muscle cells (SMCs) in the human coronary arteries and correlate their differential expression with the atherosclerosis progression. RESULTS: CD 68 and CD 34 expression increase as the atherosclerotic process proceeds from early atheroma to advanced atheroma and start to decrease as the process proceeds to fibroatheroma with a significant p < 0.001. Conversely, SMA expression decreases as the atherosclerotic process progresses with a significant p < 0.001. CONCLUSION: CD34 progenitor cells in conjunction with CD 68 macrophages have a major role in the development of atherosclerosis, whereas the SMCs are minimal in the early stages and reach their maximal levels during the stage of fibroatheroma

EFFECT OF MANGOSTEEN EXTRACT ON NEUROINFLAMMATION IN RAT MODEL OF ACUTE TRAUMATIC BRAIN INJURY

Objective: Traumatic brain injury (TBI) is one of the major health problems regarding morbidity and mortality, especially in productive ages. Following primary injury, there is a secondary insult, resulting in oxidative stress, neuroinflammation, and cell death. Mangosteen is a powerful natural antioxidant and anti-inflammation that also has neuroprotective property. The aim of this study was to explore the effect of mangosteen extract (ME)on neuroinflammation following TBI. Methods: A total of 30 Sprague-Dawley rats were randomized into three treatments group, i.e., sham-operated controls, closed head injury (CHI), and treatment group. In the treatment group, we gave ME once daily every day after CHI for 7 days. As oxidative process marker, we investigated malondialdehyde (MDA) expression. As neuroinflammation marker, we investigated glial fibrillary acidic protein (GFAP) and CD-68. Results: TBI increased the expression of GFAP and CD-68, but not MDA. There was significant GFAP expression difference between treatment group and CHI group. Regarding the expression of CD-68 and MDA, there was no significant difference between treatment and CHI group. Conclusion: Mangosteen extract supplementation decreased GFAP expression significantly after TBI

STUDY OF Coleus amboinicus STEM EXTRACT IN INHIBITING MACROPHAGE CD-68 EXPRESSION IN WISTAR RATS WITH URIC ACID-INDUCED NEPHROPATHY

The purpose of this study was to evaluate effect of Coleus amboinicus (CA) stem extracts on uric acid-induced nephropathy by comparingthe levels of Macrophage CD-68 expression and concentration of serum Cystatine C (CYS C ) in Wistar rats. Twenty-four male Wistar rats(Rattus norvegicus) with a body weight (bw) of 200-250 g, were allocated into three groups, with eight animals per group. The rats in controlgroup (PO) received 0.1% carboxymethyl cellulose (CMC) solution orally The rats in group 2 (P1) were orally induced with uric acid (UA) (500mg/kg) and oxonic acid (OA) (750 mg/kg.) and the rats in group 3 (P2) received uric acid (500 mg/kg), oxonic acid (750 mg/kg), and 500 mg/kgof the CA stem extracts for 35 days. Bloods were collected for analysis of serum CYS C expression and concentration of serum creatinine andblood nitrogen urea (BUN). The rats in all groups were sacrificed for kidney tissue extractions for macrophage CD-68 identification andhistopathology analysis. The levels of CYS C concentrations were analyzed by Avidin-Horseradish Peroxidase (HRP) Sandwich-ELISA. Theresults showed that Coleus amboinicus stem extract at dose of 500 mg/kg bw can significantly reduce BUN and creatinine levels (P≤0.05), whileCys C levels were not different. In the treatment group (P2) compared with group (P1). CD-68 (ED-1) macrophage activity decreasedsignificantly (P≤0.05) in the treatment group (P2) compared to the control group and (P1). Nephrophaty induction using UA and OA causessevere kidney lesions characterized by degeneration, necrosis and inflammation of the renal tubules and glomerulus in the treatment group

Click-modified cyclodextrins as non-viral vectors for neuronal siRNA delivery

RNA interference (RNAi) holds great promise as a strategy to further our understanding of gene function in the central nervous system (CNS) and as a therapeutic approach for neurological and neurodegenerative diseases. However, the potential for its use is hampered by the lack of siRNA delivery vectors, which are both safe and highly efficient. Cyclodextrins have been shown to be efficient and low toxicity gene delivery vectors in various cell types in vitro. However, to date they have not been exploited for delivery of oligonucleotides to neurons. To this end, a modified β-cyclodextrin (CD) vector was synthesised, which complexed siRNA to form cationic nanoparticles of less than 200nm in size. Furthermore, it conferred stability in serum to the siRNA cargo. The in vitro performance of the CD in both immortalised hypothalamic neurons and primary hippocampal neurons was evaluated. The CD facilitated high levels of intracellular delivery of labelled siRNA, whilst maintaining at least 80% cell viability. Significant gene knockdown was achieved, with a reduction in luciferase expression of up to 68% and a reduction in endogenous glyceraldehyde phosphate dehydrogenase (GAPDH) expression of up to 40%. To our knowledge, this is the first time that a modified CD has been used as a safe and efficacious vector for siRNA delivery into neuronal cells

Correlation of mRNA and protein levels: Cell type-specific gene expression of cluster designation antigens in the prostate

Background: Expression levels of mRNA and protein by cell types exhibit a range of correlations for different genes. In this study, we compared levels of mRNA abundance for several cluster designation (CD) genes determined by gene arrays using magnetic sorted and laser-capture microdissected human prostate cells with levels of expression of the respective CD proteins determined by immunohistochemical staining in the major cell types of the prostate - basal epithelial, luminal epithelial, stromal fibromuscular, and endothelial - and for prostate precursor/stem cells and prostate carcinoma cells. Immunohistochemical stains of prostate tissues from more than 50 patients were scored for informative CD antigen expression and compared with cell-type specific transcriptomes. Results: Concordance between gene and protein expression findings based on 'present' vs. 'absent' calls ranged from 46 to 68%. Correlation of expression levels was poor to moderate (Pearson correlations ranged from 0 to 0.63). Divergence between the two data types was most frequently seen for genes whose array signals exceeded background (> 50) but lacked immunoreactivity by immunostaining. This could be due to multiple factors, e.g. low levels of protein expression, technological sensitivities, sample processing, probe set definition or anatomical origin of tissue and actual biological differences between transcript and protein abundance. Conclusion: Agreement between these two very different methodologies has great implications for their respective use in both molecular studies and clinical trials employing molecular biomarkers.This work was supported by grant DK63630 and DK069690 from NIDDK. Additional funding came from grants CA85859, CA98699 and CA111244 from NCI, and PM50 GMO76547/Center for Systems Biology

Sustained Tau Phosphorylation and Microglial Activation Following Repetitive Traumatic Brain Injury

BACKGROUND: Repetitive traumatic brain injury (TBI), even without acute sequela, can induce a delayed neurodegenerative with overexpression of phosphorylated tau (p-tau) as hallmark, caused by chronic inflammation mediated in part by microglial activation. AIM: The aim of this study was to examine the dynamics of p-tau accumulation and microglial activation following repetitive TBI. MATERIALS AND METHODS: Thirty Sprague–Dawley rats were randomized into a sham control group and two treatment groups receiving three successive closed-skull impacts (TBI model) from a 40-g mass dropped from a 1-m height on alternating days (days 0, 1, 3, and 7). The first treatment group was sacrificed on the last day of trauma and the second treatment group after 7 days of no trauma. The expression level of p-tau was evaluated by AT-8 antibody immunostaining and microglial activation by anti-CD-68 immunostaining. RESULTS: Immunoexpression of AT-8 was significantly elevated 7 days after TBI compared to the last day of trauma and compared to the sham control group, while CD-68 expression was significantly higher than sham controls on the last day of trauma and remained elevated for 7 days without trauma. CONCLUSION: The study showed that brain trauma can induce p-tau overexpression and microglial activation that is sustained during the non-trauma period

1014-94 Activated Endothelial and Interstitial Cells in Chronic Myocarditis – Expression of Endothelial Adhesion Molecules

A chronic immunological process — based on an initial event of myocarditis, probably triggered by a viral infection — is considered as a possible pathogenetic mechanism for dilated cardiomyopathy (DCM). The cytokine-respondent induction of adhesion molecules on endothelial cells (EC) enables the adhesion of immunocompetent cells to activated EC and the consecutive transmigration. We studied the expression pattern of adhesion molecules (immunoglobulin-superfamily and β1-integrins) in biopsies from patients with clinically suspected DCM (n=134). Immunohistologically negative specimens (n=61: poor lymphocytic infiltration) presented a missing or weak immunoreactivity of adhesion molecules on EC. An enhanced intensity of expression was noticed in the percentage of positive biopsies (n=73: pathologically increased lymphocytic infiltration >2.0 CD 3-lymphocytes per high power field/HPF, x400-fold magnification) depicted at the following table:Negative (n=61)Positive (n=73)AntigenEndothelInterstitiumEndothelInterstitiumHLA class I15%13%63%68%HLA DR20%18%55%64%ICAM-l/CD 5425%11%84%77%VCAM-123%–88%–VLA-β/CD 2926%26%89%70%VLA-β/CDw49d15%13%66%37%LFA-3/CD 5818%16%66%36%ConclusionsPathologically increased lymphocytic infiltrates in chronic myocarditis are associated with an endothelial and interstitial inflammatory activation. This phenomenon is independent of focally concentrated infiltrates. Thus, the implication of adhesion molecules in the immunohistological diagnosis of myocarditis could provide further information apart from the sole criterion “lymphocytic infiltration” and minimize the “sampling-error-effect”

Pathologic evaluation of tumor-associated macrophage density and vessel inflammation in invasive breast carcinomas

Tumor-associated macrophages (TAMs) are major constituents of the tumor microenvironment in solid tumors and have been implicated as mediators of tumor progression, invasion and metastasis. Correspondingly, accumulation of TAMs is associated with unfavorable clinical outcomes in numerous types of solid tumors. E-selectin is a hallmark of inflammation and a key adhesion molecule that accommodates the initial contact of circulating immune cells with the inflamed vessel surface. Currently, the association between E-selectin and TAMs is not fully elucidated; therefore, the present study investigated the association between vessel inflammation, TAM infiltration, and clinical outcome in breast cancer. A total of 53 procedure-naïve invasive breast cancer cases were immunohistochemically analyzed for the presence of cluster of differentiation (CD)68+ TAMs, E-selectin+ vessels and tumor inflammation. The association between CD68 and E-selectin expression, and tumor inflammation as well as overall survival was evaluated using Kaplan-Meier survival curves and multivariable Cox\u27s proportional hazards regression analysis. The abundance of TAMs was identified to be positively associated with tumor inflammation, estrogen receptor and E-selectin expression levels. A greater prevalence of TAMs and tumor inflammation was significantly associated with shorter overall survival times. E-selectin expression levels were significantly higher in tumor vessels among elderly patients, but were not associated with overall survival. The abundance of TAMs was associated with the presence of E-selectin-expressing inflamed tumor vessels and tumor inflammation, as well as overall survival in patients with invasive breast carcinoma. © 2017, Spandidos Publications. All rights reserved

Cadmium exposure is associated with increased transcript abundance of multiple heavy metal associated transporter genes in roots of hemp (Cannabis sativa L.)

Industrial hemp (Cannabis sativa L.) has demonstrated promise for phytoremediation due to an extensive root system, large biomass, and ability to survive under relatively high levels of heavy metals. However, little research has been conducted to determine the impact of heavy metal uptake in hemp grown for medicinal use. This study evaluated the potential for cadmium (Cd) uptake and its impact on growth, physiological responses, and transcript expression of metal transporter genes in a hemp variety grown for flower production. The cultivar ‘Purple Tiger’ was exposed to 0, 2.5, 10, and 25 mg·L-1 Cd in a greenhouse hydroponic study in two independent experiments. Plants exposed to 25 mg·L-1 Cd displayed stunted plant growth characteristics, reduced photochemical efficiency, and premature senescence suggesting Cd toxicity. At the two lower concentrations of Cd (2.5 and 10 mg·L-1 Cd), plant height, biomass, and photochemical efficiency were not affected, with chlorophyll content index (CCI) being slightly lower at 10 mg·L-1 Cd, compared to 2.5 mg·L-1 Cd. There were no consistent differences between the two experiments in total cannabidiol (CDB) and tetrahydrocannabinol (THC) concentrations in flower tissues at 2.5 and 10 mg·L-1 Cd, compared to the control treatment. Root tissue accumulated the highest amount of Cd compared to other tissues for all the Cd treatments, suggesting preferential root sequestration of this heavy metal in hemp. Transcript abundance analysis of heavy metal-associated (HMA) transporter genes suggested that all seven members of this gene family are expressed in hemp, albeit with higher expression in the roots than in the leaves. In roots, CsHMA3 was up-regulated at 45 and 68 d after treatment (DAT), and CsHMA1, CsHMA4, and CsHMA5 were upregulated only under long term Cd stress at 68 DAT, at 10 mg·L-1 Cd. Results suggest that expression of multiple HMA transporter genes in the root tissue may be upregulated in hemp exposed to 10 mg·L-1 Cd in a nutrient solution. These transporters could be involved in Cd uptake in the roots via regulating its transport and sequestration, and xylem loading for long distance transport of Cd to shoot, leaf, and flower tissues

Opposite macrophage polarization in different subsets of ovarian cancer: observation from a pilot study

The role of the innate immune system in ovarian cancer is gaining importance. The relevance of tumor-associated macrophages (TAM) is insufficiently understood. In this pilot project, comprising the immunofluorescent staining of 30 biopsies taken from 24 patients with ovarian cancer, we evaluated the presence of total TAM (cluster of differentiation (CD) 68 expression), M1 (major histocompatibility complex (MHC) II expression), and M2 (anti-mannose receptor C type 1 (MRC1) expression), and the blood vessel diameter. We observed a high M1/M2 ratio in low-grade ovarian cancer compared to high-grade tumors, more total TAM and M2 in metastatic biopsies, and a further increase in total TAM and M2 at interval debulking, without beneficial effects of bevacizumab. The blood vessel diameter was indicative for M2 tumor infiltration (Spearman correlation coefficient of 0.65). These data mainly reveal an immune beneficial environment in low-grade ovarian cancer in contrast to high-grade serous ovarian cancer, where immune suppression is not altered by neoadjuvant therapy