Leukocyte Apoptosis Research Articles

Varis etiyopatogenezinde lökosit aktivasyonu ile apopitozisin ilişkisi

Aim: Varicose veins are the most common disorders of the lower extremities. The purpose of this study is to investigate the effect of leukocyte activation in the vein wall and apoptosis in the etiopathogenesis of varicose veins.Material and Methods:Forty-six patients with varicose veins were included in the study. High ligation with stripping with or without additional individual ligation and excision was performed for symptoms, complications or cosmetic needs. ELISA was used to measure the serum concentration of caspase-8 and caspase-9.Results: There was a statistically significant difference between the cephalic vein and saphenous vein groups (p=0.04 and p<0.0001 respectively). Conclusion: As a conclusion, the complex pathophysiology underlying varicose veins has yet to be fully defined. It is suggested that both leukocyte activation and dysregulation of apoptosis are associated with susceptibility for varicose vein formation however this argument needs further research.

Targeted Delivery of Curcumin Rescues Endoplasmic Reticulum-Retained Mutant NOX2 Protein and Avoids Leukocyte Apoptosis.

Chronic granulomatous disease (CGD) is a primary immunodeficiency disease caused by defects in the leukocyte NADP oxidase. We previously reported that sarcoplasmic/endoplasmic reticulum calcium pump (SERCA) inhibitors could be used to rescue mutant H338Y-gp91phox protein of a particular type of CGD with a CybbC1024T mutation, leading to endoplasmic reticulum (ER) retention of the mutant protein. In this study, we developed a novel mouse model with the CybbC1024T mutation on a Cybb knockout background and investigated the therapeutic effects of ER-targeted delivery of the SERCA inhibitor, curcumin, with poly(lactic-coglycolic acid) (PLGA) nanoparticles (NPs). We found that PLGA encapsulation improved the efficacy of curcumin as a SERCA inhibitor to induce ER calcium release. ER-targeting curcumin-loaded PLGA NPs reduced and delayed extracellular calcium entry and protected the cells from mitochondrial damage and apoptosis. In vivo studies showed that ER-targeting curcumin-loaded PLGA NPs treatment enhanced neutrophil gp91phox expression, ROS production and peritoneal bacterial clearance ability of the CybbC1024T transgenic Cybb -/- mice. Our findings indicate that ER-targeted delivery of curcumin not only rescues ER-retained H338Y-gp91phox protein, and hence leukocyte function, but also enhances the bioavailability and reduces cytotoxicity. Modulation of ER function by using organelle-targeted NPs may be a promising strategy to improve the therapeutic potential of curcumin as a treatment for CGD.

Methylmercury, cadmium and arsenic(III)-induced toxicity, oxidative stress and apoptosis in Pacific red snapper leukocytes

Methylmercury (MeHg), cadmium (Cd) and arsenic (As(III)) are among the most toxic metals in aquatic systems that have been associated with multiple animal and human health problems. This study investigated cytotoxic, oxidative stress, and apoptosis effects on fish leukocytes following their exposure to metals. A preliminary study indicated that leukocytes exposed to MeHg at a concentration of 0.01 mM, Cd at 0.05 mM, and As(III) at 2 mM showed a time-dependent cell viability reduction (around 40%), so they were selected for further experiments. To evaluate the effect of MeHg, Cd and As(III) on Pacific red snapper Lutjanus peru, we measured cytotoxicity, reactive oxygen species, antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT)), nitric oxide production, apoptosis-related and immune-related genes on head-kidney and spleen leukocytes following exposure to MeHg (0.01 mM), Cd (0.05 mM) and As(III) (2 mM) for 30 min and 2 h. Reactive oxygen species (ROS) generation highly increased in time-dependent doses in head-kidney leukocytes compared with the control group. Regarding antioxidant activity, SOD increased significantly in leukocytes exposed to any heavy metals after two h. Expressly, CAT activity decreased in those leukocytes exposed to Cd and As(III). Apoptotic function genes (Casp-2, Casp-3, and Casp-7) strongly up-regulated after heavy metal exposure, but Cd was more toxic. Finally, granzyme A and perforin 1 strongly up-regulated in leukocytes exposed to MeHg and As(III) compared with the control group. Our data showed that MeHg, Cd, and As(III) might have been cytotoxic and induced oxidative stress and apoptosis with possible biological consequences in fish.

Study of cell death kinds and stages of leukocytes apoptosis in pulmonary tuberculosis patient during antimycobacterial therapy

Study of cell death kinds and stages of leukocytes apoptosis in pulmonary tuberculosis patient during antimycobacterial therapy

Curcumin induced oxidative stress causes autophagy and apoptosis in bovine leucocytes transformed by Theileria annulata

Bovine tropical theileriosis is a tick-borne disease, caused by Theileria annulata which is a protozoan parasite that resides within the B-cells and macrophages. T. annulata is a unique parasite that can transform bovine leucocytes which leads to the cancer hallmarks in the infected cells. Previously, curcumin has been shown to possess multiple pharmacological activities such as anti-inflammatory and anti-cancer activities. In this study, we demonstrated that curcumin inhibits the proliferation of Theileria-transformed bovine leucocytes by promoting apoptosis and autophagy. The transcriptome analysis of curcumin treated cells showed that the genes involved in cell death and autophagy are also differentially regulated. We further elucidated the mechanism of action of curcumin on Theileria infected bovine cells. We found that curcumin induced the generation of reactive oxygen species (ROS) which activated caspase 8 and destabilized the mitochondrial membrane potential leading to the release of cytochrome c from mitochondria. This subsequently led to the activation of caspase 3 and PARP cleavage, finally leading to apoptosis in the infected cells. Furthermore, curcumin induced the process of autophagy which was characterized by the formation of acidic vesicular organelles, LC3B accumulation with lysosome inhibitor, E64d, and the presence of autophagosomes as visualized by transmission electron microscopy (TEM). Curcumin treatment suppressed the mTOR and increased the expression of autophagy-related proteins. We also found that N- acetylcysteine, an inhibitor of ROS, could rescue the infected cells from curcumin induced apoptosis and autophagy mediated cell death. Intriguingly, curcumin had no effect on uninfected bovine PBMCs. Altogether, these data suggest the therapeutic potential of curcumin against bovine tropical theileriosis.

Therapeutic effects of a phospholipase D1 inhibitor against polymicrobial sepsis

Abstract In this study, we found that a phospholipase D1-selective inhibitor (VU0155069), significantly increased survival rate against cecal ligation and puncture (CLP)-induced sepsis. Pathological events caused by CLP such as lung inflammation and widespread leukocyte apoptosis were markedly inhibited by the PLD1 inhibitor. The PLD1 inhibitor also strongly inhibited the production of proinflammatory cytokines especially IL-1β in CLP mice and in LPS-stimulated splenocytes. VU0155069 also significantly blocked IL-1β production, caspase-1 activation, and pyroptosis caused by several inflammasome-activating signals (LPS+MSU, NIG, dsDNA) in mouse bone marrow-derived macrophages (BMDM). However, VU0155069 did not affect LPS-induced activation of signaling such as MAPK (ERK, p38 MAPK, JNK), Akt, NF-kB, and NLRP3 expression in BMDM. The PLD1 inhibitor also failed to affect mitochondrial ROS generation and calcium increase caused by LPS or ATP, and subsequent ASC oligomerization caused by several inflammasome-activating signals. The PLD1 inhibitor inhibited caspase-1 activity caused by LPS+NIG in BMDM, which is not a direct effect on caspase-1. Our results suggest that the PLD1 inhibitor shows anti-septic activity as well as inflammasome-inhibiting effects.

Влияние цитофлавина на апоптоз лейкоцитов крови пациентов с ишемической болезнью сердца в зависимости от носительства полиморфизма гена цитохрома CYP2C9 в условиях <i>in vitro</i>

С целью определения характера влияния цитофлавина на апоптоз лейкоцитов крови пациентов с ишемической болезнью сердца (ИБС) в зависимости от наличия полиморфизма гена цитохрома CYP2C9*2 C430T были обследованы 90 пациентов с ИБС: стабильной стенокардией напряжения. Методом полимеразной цепной реакции определяли генные полиморфизмы цитохрома CYP2C9*2 C430T. Методом ДНК-комет, который позволяет оценить уровень апоптоза белых клеток крови, по разработанному авторами способу в тестах in vitro оценивали влияние цитофлавина в терапевтической концентрации на ДНК лейкоцитов крови больных. При частотном анализе гена CYP2C9*2 C430T среди пациентов было установлено, что 75 человек (83 %) гомозиготны по дикому аллелю (генотип СС), 15 человек (17 %) — гетерозиготны (генотип СТ), гомозиготных лиц по полиморфному аллелю (генотип ТТ) в выборке не обнаружено. У носителей полиморфного гена цитохрома CYP2C9*2 С430Т (генотипа СТ) наблюдали более сильное повреждение ДНК в исходном статусе (11,00 ± 3,21) % повреждённых клеток в сравнении с (3,43 ± 1,13) % у пациентов с генотипом СС (р = 0,02). При введении цитофлавина в терапевтической концентрации in vitro обнаружили тенденцию разнонаправленного изменения состояния ДНК лейкоцитов в зависимости от наличия полиморфизма гена CYP2C9*2 С430Т. Так, у пациентов с диким геном CYP2C9*2 С430Т при введении цитофлавина in vitro обнаружили незначительное недостоверное увеличение количества апоптотических лейкоцитов крови, в то время как у пациентов с полиморфным геном CYP2C9*2 С430Т (генотипом СТ) обнаружили чёткую тенденцию снижения количества апоптотических клеток практически до нулевых значений показателя (p = 0,087), что позволяет выдвинуть гипотезу о возможной способности цитофлавина модулировать стабильность ДНК в зависимости от состояния генома и о необходимости разработки персонализированных подходов к использованию цитофлавина у пациентов с ИБС в клинической практике.

Razlikovanje modaliteta ćelijske smrti leukocita korišćenjem protočne citometrije kod bolesnika sa hroničnim rinosinuzitisom bez nazalnih polipa

Objective. The objective of our research is to study the rate and modes of cell death among white blood cells using an Annexin V and 7-AAD flow cytometry kit in patients with chronic rhinosinusitis without nasal polyps (CRSsNP). Methods. Flow cytometry was used to detect cell death modes of leukocytes in twelve patients with CRSsNP. Annexin V FITC, anti-CD45 PE and 7-AAD (7-aminoactinomycin D) were used for the assessment of apoptosis stages. The current method allows identifying four different states of cells: 1 - viable cells (Annexin Vnegative,7-AAD negative cells); 2 - early apoptotic cells (AnnexinV positive, 7-AAD negative cells); 3 - late apoptotic/necrotic cells (AnnexinV positive, 7-AAD positive cells); 4 - dead necrotic cells (AnnexinV negative, 7-AAD positive cells). To assess the rate of oxidative stress in CRSsNP, 8-isoprostane levels in blood serum were determined using ELISA. Results. We found that CRSsNP was accompanied by the development of oxidative stress, evidenced by elevation of 8isoprostane in blood serum, and the increased rate of cell death among white blood cells with the prevalence of early apoptotic cells, confirmed by a higher percentage of AnnexinV positive, 7-AAD negative cells compared with control subjects. Conclusion. Our findings suggest that CRSsNP is accompanied by the activation of leukocyte apoptosis

Redox correlation in muscle lengthening and immune response in eccentric exercise.

This study was designed to examine the potential involvement of reactive oxygen species in skeletal muscle dysfunction linked with stretching in a mouse model and to explore the effects of combined antioxidant intake on peripheral leukocyte apoptosis following eccentrically-biased downhill runs in human subjects. In the mouse model, diaphragmatic muscle was stretched by 30% of its optimal length, followed by 5-min contraction. Muscle function and extracellular reactive oxygen species release was measured ex vivo. In human models, participants performed two trials of downhill running either with or without antioxidant supplementation, followed by apoptotic assay of inflammatory cells in the blood. The results showed that stretch led to decreased muscle function and prominent ROS increase during muscle contraction. In human models, we observed an elevation in circulating leukocyte apoptosis 24–48 hours following acute downhill runs. However, there is an attenuated leukocyte apoptosis following the second bout of downhill run. Interestingly, the combination of ascorbic acid (vitamin C) and α-tocopherol (vitamin E) supplementation attenuated the decrease in B-cell lymphoma 2 (Bcl-2) at 24 hours following acute downhill running. These data collectively suggest that significant ROS formation can be induced by muscle-lengthening associated with eccentric exercise, which is accompanied by compromised muscle function. The combination of antioxidants supplementation appears to have a protective role via the attenuation of decrease in anti-apoptotic protein.

L-arginine inhibited apoptosis of fish leukocytes via regulation of NF-κB-mediated inflammation, NO synthesis, and anti-oxidant capacity

The increased apoptosis plays an important role in bacterial invasion. In addition, LPS can induce inflammation and apoptosis of leukocytes via the production of reactive oxygen and nitrogen species. In the present study, we investigated the potential protective role of l-arginine (L-Arg) against the apoptosis of fish leukocytes in vitro. The results of Annexin V-FITC/PI staining and TUNEL assay indicated that L-Arg significantly alleviated the apoptosis of fish leukocytes induced by LPS at 24 h and 72 h post incubation (hpi). High caspase-3 activities induced by LPS at 72 hpi were significantly inhibited by L-Arg. Moreover, L-Arg supplementation also significantly decreased the mRNA expression levels of caspases at most time points, which contributed to the anti-apoptotic roles of L-Arg. Further analysis showed that L-Arg significantly inhibited the expression of several pro-inflammatory cytokines including IL-8 and TNF-α, partially via the down-regulation of the genes involved in NF-κB/MyD88 including NF-κB, IKKα and IKKγ. The down-regulation of these pro-inflammatory cytokines by L-Arg supplementation led to the further decrease in the expression of death receptor FasL, contributing to the anti-apoptotic effect of L-Arg. In addition, L-Arg supplementation increased both iNOS mRNA expression and NO production. The mRNA expressions of several anti-oxidant enzymes including SOD, CAT and GSHPx were also significantly increased after L-Arg supplementation, which accelerated the clearance of reactive oxygen species. In all, L-Arg inhibited apoptosis of fish leukocytes both via the increased NO production and antioxidant capacity and via the inhibition of inflammation mediated by NF-κB/MyD88 pathway.

Analysis of differential gene expression by RNA-seq data in ABCG1 knockout mice

AimsThe previous studies on ABCG1 using genetically modified mice showed inconsistent results on atherosclerosis. The aim of this study was to determine whether accurate target knockout of ABCG1 would result in transcriptional changes of other atherosclerosis-related genes. MethodsABCG1 knockout mouse model was obtained by precise gene targeting without affecting non-target DNA sequences in C57BL/6 background. The wildtype C57BL/6 mice were regarded as control group. 12-week-old male mice were used in current study. We performed whole transcriptome analysis on the peripheral blood mononuclear cells obtained from ABCG1 knockout mice (n = 3) and their wildtype controls (n = 3) by RNA-seq. ResultsCompared with wildtype group, 605 genes were modified at the time of ABCG1 knockout and expressed differentially in knockout group, including 306 up-regulated genes and 299 down-regulated genes. 54 genes were associated with metabolism regulation, of which 13 were related to lipid metabolism. We also found some other modified genes in knockout mice involved in cell adhesion, leukocyte transendothelial migration and apoptosis, which might also play roles in the process of atherosclerosis. 7 significantly enriched GO terms and 19 significantly enriched KEGG pathways were identified, involving fatty acid biosynthesis, immune response and intracellular signal transduction. ConclusionsABCG1 knockout mice exhibited an altered expression of multiple genes related to many aspects of atherosclerosis, which might affect the further studies to insight into the effect of ABCG1 on atherosclerosis with this animal model.

Kidney histopathology in lethal human sepsis

PurposeThe histopathology of sepsis-associated acute kidney injury (AKI) in critically ill patients remains an understudied area. Previous studies have identified that acute tubular necrosis (ATN) is not the only driver of sepsis-AKI. The focus of this study was to identify additional candidate processes that may drive sepsis-AKI. To do this we immunohistochemically characterized the histopathological and cellular features in various compartments of human septic kidneys.MethodsWe studied the following histopathological features: leukocyte subsets, fibroblast activation, cellular proliferation, apoptosis, and fibrin deposition in the glomerulus and the tubulointerstitium in human post-mortem kidney biopsy tissue. Biopsy tissue samples from 27 patients with sepsis-AKI were collected 33 min (range 24–150) after death in the ICU. The unaffected part of the kidneys from 12 patients undergoing total nephrectomy as a result of renal carcinoma served as controls.ResultsImmunohistochemical analysis revealed the presence of more neutrophils and macrophages in the glomeruli and more neutrophils in the tubulointerstitium of renal tissue from patients with sepsis compared to control renal tissue. Type II macrophages were predominant, with some macrophages expressing both type I and type II markers. In contrast, there were almost no macrophages found in control kidneys. The number of activated (myo)fibroblasts was low in the glomeruli of sepsis-AKI kidneys, yet this was not observed in the tubulointerstitium. Cell proliferation and fibrin deposition were more pronounced in the glomeruli and tubulointerstitium of sepsis-AKI than in control kidneys.ConclusionsThe extensive heterogeneity of observations among and within patients emphasizes the need to thoroughly characterize patients with sepsis-AKI in a large sample of renal biopsy tissue from patients with sepsis.

Abnormal microRNA expression in the course of hematological malignancies.

Research on the carcinogenesis process is currently focused primarily on understanding its genetic basis and molecular abnormalities that may be predictive factors and therapeutic targets. It was clearly confirmed recently that microRNAs are involved in the mechanisms of leukocyte development, differentiation, and apoptosis, as well as in the pathogenesis of proliferative diseases of the hematopoietic system. Currently, research strategies allow determination of the deregulation of microRNA profiles in relation to other cytogenetic aberrations, as well as prognostic factors and primary end points. The problem of the possibility of their use as therapeutic targets is also increasingly discussed. In this article, we analyze literature data on abnormalities in microRNA expression in proliferative diseases of the hematopoietic system in the context of classic cytogenetic and molecular aberrations.

Interaction of systemic oxidative stress and mesial temporal network degeneration in Parkinson\u2019s disease with and without cognitive impairment

BackgroundTo identify the vulnerable areas associated with systemic oxidative stress and further disruption of these vulnerable areas by measuring the associated morphology and functional network alterations in Parkinson’s disease (PD) patients with and without cognitive impairment.MethodsThis prospective study was approved by the institutional review board of KCGMH, and written informed consent was obtained. Between December 2010 and May 2015, 41 PD patients with different levels of cognitive functions and 29 healthy volunteers underwent peripheral blood sampling to quantify systemic oxidative stress, as well as T1W volumetric and resting state functional MRI (rs-fMRI) scans. Rs-fMRI was used to derive the healthy intrinsic connectivity patterns seeded by the vulnerable areas associated with any of the significant oxidative stress markers. The two groups were compared in terms of the functional connectivity correlation coefficient (fc-CC) and gray matter volume (GMV) of the network seeded by the vulnerable areas.ResultsThe levels of oxidative stress markers, including leukocyte apoptosis and adhesion molecules, were significantly higher in the PD group. Using whole-brain VBM-based correlation analysis, the bilateral mesial temporal lobes (MTLs) were identified as the most vulnerable areas associated with lymphocyte apoptosis (P < 0.005). We found that the MTL network of healthy subjects resembled the PD-associated atrophy pattern. Furthermore, reduced fc-CC and GMV were further associated with the aggravated cognitive impairment.ConclusionThe MTLs are the vulnerable areas associated with peripheral lymphocyte infiltration, and disruptions of the MTL functional network in both architecture and functional connectivity might result in cognitive impairments in Parkinson’s disease.

Abstract B63: Bittersweet symphony: How tumor-associated glycan structures orchestrate immune evasion

Abstract The tumor microenvironment is immune suppressive, allowing tumor cells to escape from immune attack. By relieving this immune suppression, immune cells might be able to effectively eliminate the tumor. Therefore, a detailed understanding on how tumor cells induce an immunosuppressive tumor microenvironment is required. Glycosylation is the most abundant post-translational modification of proteins. It is highly diverse on a cellular level and strongly affected by oncogenesis. Tumor cells generally display an aberrant glycosylation profile and this is now well accepted as a new hallmark of cancer. Overexpression of glycan structures containing sialic acids (Sia) is frequently observed on tumors. Glycan structures can be recognized by lectin receptors expressed on immune cells, whereby sialic acids specifically bind to the Siglec receptor family. The majority of Siglec receptors has an immunoreceptor tyrosine-based inhibition motif and therefore, sialylated glycan structures are known to suppress ongoing immune responses. Hence, we hypothesize that tumor cells overexpress sialylated glycan structures in order to communicate with the immune system and to dampen immune attack. We have previously demonstrated that a genetically engineered Sialow glycovariant of the mouse melanoma B16 model displayed delayed growth in vivo compared to wild-type B16 Siahigh tumors (1). Moreover, higher frequencies of IFNγ-producing effector T-cells were detected in B16 Sialow tumors. This was paralleled by a 50% reduction in FoxP3+CD4+ regulatory T-cell frequencies in Sialow tumors (1), together indicating that tumor-associated sialic acids indeed play, indeed, a crucial role in suppressing the anti-tumor immune response. To check whether the inhibitory effect of sialic acids on the effector T cell response is tumor type specific, we investigated the role of sialic acids in a mouse colorectal cancer model (MC38). With the use of Crispr-Cas9 we successfully knocked out the N-acylneuraminate cytidylyltransferase (CMAS) gene, which is responsible for the sialylation pathway and thus the generation of sialylated glycans on the cell surface. In contrast to the B16 Sialow tumors, the Sianeg MC38 CMAS KO tumors displayed an increased tumor growth in vivo compared to the Siahigh MC38 Mock tumors. Moreover, less viable leukocytes could be recovered from the Sianeg MC38 CMAS KO tumors compared to the Siahigh MC38 Mock tumors. With the use of mass spectrometry, we analyzed the complete glycosylation profile of our Sianeg MC38 CMAS KO and Siahigh MC38 Mock cells and observed, in addition to the absence of sialylation, an increase in LacNAc (Galβ1-4GlcNAc) structures on the Sianeg MC38 CMAS KO cells. This glycan structure is a ligand for carbohydrate-binding Galectin receptors. Galectins are well known for their potential to induce leukocyte apoptosis after binding specific glycoproteins on the leukocyte surface. Using qPCR we observed higher mRNA levels of Galectin-1 in our MC38 CMAS KO cells. The enhanced production of Galectin-1 and the elevated expression of LacNAc together suggest that MC38 CMAS KO cells are able to recruit Galectin-1, thereby facilitating the enhanced immune suppression in these tumors. Currently, we are confirming the role of Galectins in our MC38 CMAS KO tumors. In conclusion, tumor cells alter their glycosylation profile and based on our data, we propose that tumor cells use these glycan structures as a mechanism to influence antitumor immunity and that this phenomenon might be tumor-type specific. Their immune inhibitory nature and their tumor-specific expression identifies tumor-associated glycans as potent candidates for future immune checkpoint blockade therapy. Reference: 1. Perdicchio M. et al. Tumor sialylation impedes T cell mediated anti-tumor responses while promoting tumor associated-regulatory T cells. Oncotarget 2016;7(8):8771-8. Citation Format: Lenneke A.M. Cornelissen, Athanasios Blanas, Joost C. Van der Horst, Laura Kruijssen, Yvette Van Kooyk, Sandra J. Van Vliet. Bittersweet symphony: How tumor-associated glycan structures orchestrate immune evasion [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2017 Oct 1-4; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2018;6(9 Suppl):Abstract nr B63.

In Vivo genotoxicity of a commercial C.I. Disperse Red 1 dye.

Color Index (C.I.) Disperse Red 1 (DR1) is a widely used textile azo dye found in rivers. As it may not be completely removed by conventional treatments, humans can be exposed through drinking water. Studies have supported in vitro toxicity and mutagenicity of commercial DR1. This study aimed to investigate the mutagenic and toxicogenomic effects of commercial DR1 in multiple tissues/organs of Swiss male mice. For that, animals were orally exposed to the dye (by gavage), at single doses of 0.0005, 0.005, 0.5, 50, or 500 mg/kg bw. The two lowest doses were equivalent to the ones found in two Brazilian rivers receiving influx of textile discharges. Cytotoxicity, micronucleated cell frequencies (for all doses tested), primary DNA damage (comet assay), and gene expression profiling of (0.0005 and 0.005 mg/kg of bw) were investigated 24 hr after animal exposure to commercial DR1. Data showed increased frequencies of micronucleated polychromatic erythrocytes in bone marrow cells after treatment with 0.5 and 50 mg/kg bw. At 0.005 mg/kg bw, commercial DR1 induced an increase of primary DNA damage in liver, but not in kidney cells. Additionally, upregulation of genes involved in the inflammatory process (IL1B) (0.0005 and 0.005 mg/kg bw) and cell-cycle control (CDKN1A) in liver cells, and apoptosis (BCL2 and BAX) in leukocytes (0.005 mg/kg bw) were also detected. In conclusion, the commercial DR1 was genotoxic (chromosome aberrations and primary DNA damage) and modulated gene expression in mice, and such effects were dependent on the doses and tissues analyzed. Environ. Mol. Mutagen. 59:822-828, 2018. © 2018 Wiley Periodicals, Inc.

Functional convergence of gliP and aspf1 in Aspergillus fumigatus pathogenicity

ABSTRACT Gliotoxin contributes to the virulence of the fungus Aspergillus fumigatus in non-neutropenic mice that are immunosuppressed with corticosteroids. To investigate how the absence of gliotoxin affects both the fungus and the host, we used a nanoString nCounter to analyze their transcriptional responses during pulmonary infection of a non-neutropenic host with a gliotoxin-deficient ΔgliP mutant. We found that the ΔgliP mutation led to increased expression of aspf1, which specifies a secreted ribotoxin. Prior studies have shown that aspf1, like gliP, is not required for virulence in a neutropenic infection model, but its role in a non-neutropenic infection model has not been fully investigated. To investigate the functional significance of this up-regulation of aspf1, a Δaspf1 single mutant and a Δaspf1 ΔgliP double mutant were constructed. Both Δaspf1 and ΔgliP single mutants had reduced lethality in non-neutropenic mice, and a Δaspf1 ΔgliP double mutant had a greater reduction in lethality than either single mutant. Analysis of mice infected with these mutants indicated that the presence of gliP is associated with massive apoptosis of leukocytes at the foci of infection and inhibition of chemokine production. Also, the combination of gliP and aspf1 is associated with suppression of CXCL1 chemokine expression. Thus, aspf1 contributes to A. fumigatus pathogenicity in non-neutropenic mice and its up-regulation in the ΔgliP mutant may partially compensate for the absence of gliotoxin. Abbreviations:PAS: periodic acid-Schiff; PBS: phosphate buffered saline; ROS: reactive oxygen species; TUNEL: terminal deoxynucleotidyl transferase dUTP nick-end labeling

Examination of the cellular mechanisms of leukocyte elevation by 10.6\xa0\u03bcm and 650\xa0nm laser acupuncture-moxibustion

To investigate the cell cycle and cellular mechanisms of leukocyte elevation by laser acupuncture in rats with cyclophosphamide (CTX)-induced leukopenia. Sixty-six rats were randomized into six groups: normal, model control group, sham treatment group, 10.6 μm laser treatment group, 650 nm laser treatment group, and 10.6 μm–650 nm compound laser treatment group. Eleven rats were used in the normal group and 55 were models that were injected with cyclophosphamide to induce leukopenia. For the three laser treatment groups, 10.6-μm and 650-nm lasers, and 10.6-μm–650-nm compound lasers were used to irradiate the DU14 (Dazhui) and bilateral ST36 (Zusanli) for 5 min each. The sham laser group received the same operation as the laser group but without irradiation. The normal group and model group were not treated. Differences in the number of nucleated cells in the femoral bone marrow, and cell cycle and cellular apoptosis of peripheral leukocytes in rats in various groups were compared. Compared with the model group and the sham laser group, the number of nucleated cells in the femoral bone marrow in the 10.6-μm laser, 650-nm laser, and 10.6-μm–650-nm compound laser group was significantly increased after treatment (P = 0.001, 0.002, 0.034, respectively) and did not show any significant difference with the normal group (P = 0.964, P = 0.838, P = 0.287, respectively). The number of cells in G2 phase in the 10.6 μm laser group was similar to that of the normal group (P = 0.973). The number of cells in G2 phase in the model, sham, 650-nm laser group, and 10.6-μm–650-nm compound laser group were significantly lower than in the normal group and 10.6-μm laser group (P = 0.016, P = 0.023, P = 0.044, P = 0.039, respectively). In the model group and the sham treatment group, the apoptosis rates of peripheral leukocytes were increased compared with the normal group (P = 0.001), while the proportion of cells in the G2 phase was significantly lower than in the normal group (P = 0.016), and the proportion of cells in S phase was higher than in the normal group (P = 0.014). The incidence of apoptosis in peripheral blood cells in the three laser treatment groups did not show any statistically significant difference when compared with the normal group (P > 0.05). Treatment with the 10.6-μm, 650-nm, and 10.6-μm–650-nm compound lasers increased the number of nucleated cells in the bone marrow, decreased the unfavorable effects of cyclophosphamide on the cell cycle, induced the cell cycle towards proliferation, decreased apoptosis, improved the intramedullary hematopoietic system, and increased peripheral leukocyte count.

Glucocorticoids, Sex Hormones, and Immunity.

Glucocorticoid hormones regulate essential body functions in mammals, control cell metabolism, growth, differentiation, and apoptosis. Importantly, they are potent suppressors of inflammation, and multiple immune-modulatory mechanisms involving leukocyte apoptosis, differentiation, and cytokine production have been described. Due to their potent anti-inflammatory and immune-suppressive activity, synthetic glucocorticoids (GCs) are the most prescribed drugs used for treatment of autoimmune and inflammatory diseases. It is long been noted that males and females exhibit differences in the prevalence in several autoimmune diseases (AD). This can be due to the role of sexual hormones in regulation of the immune responses, acting through their endogenous nuclear receptors to mediate gene expression and generate unique gender-specific cellular environments. Given the fact that GCs are the primary physiological anti-inflammatory hormones, and that sex hormones may also exert immune-modulatory functions, the link between GCs and sex hormones may exist. Understanding the nature of this possible crosstalk is important to unravel the reason of sexual disparity in AD and to carefully prescribe these drugs for the treatment of inflammatory diseases. In this review, we discuss similarities and differences between the effects of sex hormones and GCs on the immune system, to highlight possible axes of functional interaction.

Transcriptome analysis provides insights into the immunity function of venom glands in Pardosa pseudoannulata in responses to cadmium toxicity.

Due to some similarity of innate immunity between insects and mammals, the study of the molecular mechanism of innate immunity in insects has become a focus of research. However, the exact molecular and cellular basis of immune system in insect remains poorly understood. Characterization of the transcriptomic response to Cd of spider is an effective approach to understanding the innate immunity mechanisms. In this study, we carried out transcriptome sequencing and gene expression analyses to develop molecular resources for Pardosa pseudoannulata venom glands with and without Cd treatments. A total of 92,778 assembled unigenes and 237 Cd stress-associated differentially expressed genes between the Cd-treated and control groups were obtained. Expression profile analysis demonstrated that immunity-related genes involved in bacterial invasion of epithelial cells, leukocyte transendothelial migration, platelet activation, apoptosis, phagosome, and Rap1 signaling pathway were upregulated by Cd exposure, except the genes involved in PPAR signaling pathway were downregulated. Our results provide the first comprehensive transcriptome dataset of venom glands in P. pseudoannulata response to Cd, which is valuable for throws light on the immunotoxicity mechanism of Cd, and the innate immunity complexity.