Beta Gene Expression Research Articles

Molecular Interplay of John Cunningham Virus with Interleukin 1 Beta in Colorectal Carcinomatous Tissues from a Group of Iraqi Patients.

Colorectal cancer is ranked to have high mortality among most malignancies worldwide. In the adult population, the seroprevalence rates of the John Cunningham virus (JCV) range from 70% to 90%. Recently the association for JCV in many malignant tumours have been reported worldwide, including colonic and rectum cancers. Interleukin-1β (IL-1β) can promote tumour growth where it is abundant in the tumour microenvironment, and its up-regulation is considered a poor prognostic feature in different types of solid tumours, including colon malignancies. One hundred tissue biopsies belonged to 50 patients with colorectal cancers and 30 benign colonic tumour patients, and 20 colorectal control tissues were enrolled in this study. JCV was detected via chromogenic in situ hybridization (CISH), while IL-1 beta was detected by immunohistochemistry (IHC). The recorded data showed that 21 out of 50 (42%) tissue samples with colorectal carcinoma showed positive CISH reactions for JCV DNA in this study. The benign colorectal tumours group revealed positive signals in 2 out of 30 tissues representing 6.7% of this group. Lastly, no control tissues showed positive signals for the JCV -CISH test. The positive signals of IL-1 Beta-IHC detection were found in 26 out of 50 (52 %) colorectal carcinoma tissues, while in the benign colorectal tumour was 43.3% (13 out of 30) and in AHC was 20% (4 out of 20 tissues). The high rates of JCV infection in this group of Iraqi patients with colonic adenocarcinoma in concordance with IL-1 Beta expression could play an essential role in the development and progression of these malignant tumours along with benign colonic tumours. To analyze the concordant expression of IL-1 beta gene and JCV in issues from a group of Iraqi patients with colonic adenocarcinomas.

Evaluation of tooth eruption rate of incisor teeth in rats with estrogen deficiency.

To assess the influence of estrogen deficiency on tooth eruption rate (TER) and gene expression of estrogen receptor alpha and beta (ERα and ERβ) in the odontogenic region of teeth with continuous formation in a rat model. Ovariectomies (OVX; n = 25) and sham surgeries (SHAM; n = 25) were performed in female Wistar rats when animals were 25days old. The TER of the lower incisors, both in impeded (hyperfunction condition) and unimpeded (trimmed incisal edge-hypofunction condition) conditions, was evaluated using standardized digital photographs acquired every 48-72h for 3weeks (35th-53rd day of life), using a camera coupled to a stereomicroscope. Quantitative real-time PCR was performed to evaluate the relative gene expression of ERα and ERβ in the odontogenic region. The OVX group showed a significant reduction in TER when compared to the SHAM group, only in the impeded condition (p = 0.03). There was no statistically significant difference between the groups in ERα gene expression (p = 0.33). ERβ showed a significantly higher gene expression in the OVX group (p ≤ 0.05). Estrogen deficiency decreases TER in teeth under impeded condition. Estrogen deficiency also increases ERβ gene expression in the odontogenic region of teeth with continuous formation. Hormonal disturbances affecting estrogen levels can cause alterations in dental formation and teeth eruption.

Expression of IL1β Gene in the Placentas from Humans and Cows during Brucella Infection.

Brucellosis is an important contagious disease affecting most domestic and mature animals. Since the impact of IL-1β in B. abortus invasion and survival remains elusive, the current study sought to elucidate the actual roles of these potent cytokines in the modulation of the initial immune response to Brucella infection. Therefore, this study aimed to detect Brucella abortus in the placenta of aborted women and cows and estimate the expression of the interleukin 1β (IL1β) gene associated with immune response mechanisms to Brucella abortus infection. The detection of Brucella abortus was performed by Rose Bengal Test (RBT) and Polymerase Chain Reaction based AlkB gene (AlkB-PCR) in the sera and placenta samples of aborted women and cows, respectively. The overall percentage of Brucella abortus infection was 13.1% and 5% as determined by RBT and AlkB-PCR in aborted women's sera and placentas, respectively. On the other hand, the overall percentage rates of Brucella abortus infection in the sera and placentas from aborted cows were 30% and 11% as estimated by RBT and AlkB-PCR, respectively. The results of RBT demonstrated that the association between Brucella abortus and abortion in cows was statistically significant. On the other hand, it was found that the association between Brucella abortus and abortion in women was not significant. Moreover, according to the results of AlkB-based PCR, the association between Brucella abortus and abortion was statistically significant in aborted cows, while it was not significant in aborted women. The sensitivity, specificity, and accuracy of RBT were calculated as 60.00, 53.85, and 54.55%, respectively. Moreover, positive and negative predictive values were reported as 14.33% and 91.28%, respectively. Regarding RBT for aborted cows, the sensitivity, specificity, and accuracy of the test were 81.82%, 57.78%, and 62.49%, respectively. The positive predictive value was reported as 32.08%, while the negative predictive value was reported as 92.88%. Quantitative PCR (qPCR) was carried out for the evaluation of Interleukin 1 Beta (IL1β) gene expression. The qPCR result was presented as a fold change in gene expression. A significant increment of IL1β gene expression was observed in aborted women (114.905±99.661) and cows (22.454 ±18.528), compared to non-aborted women (4.953±5.564) and cows (2.033±1.845). Statistical comparison of IL1β gene expression between aborted women and cows illustrated a non-significant increment in IL1β gene expression in aborted women (114.905±99.661), compared to aborted cows (22.454 ±18.528).

Investigating histological changes and gene expression of interleukin-8, interleukin-12, and interferon beta in liver tissue after consumption of Lactobacillus rhamnosus GG probiotic in steatosis model rats

Investigating histological changes and gene expression of interleukin-8, interleukin-12, and interferon beta in liver tissue after consumption of Lactobacillus rhamnosus GG probiotic in steatosis model rats

Structural, optical characteristics and Anti-Cancer effect of Cd0.99Ni0.01O nanoparticles on human neuroblastoma and cervical cancer cell lines

Cd0.99Ni0.01O nanoparticles are outstanding nanomaterials used in different applications. So the structural, morphological, and optical properties of these nanoparticles are employed in anticancer applications. X-ray diffraction data shows the single phase with polycrystalline nature. The structural parameters were calculated from the MAUD program. Also, the TEM confirmed the size distribution of Cd0.99Ni0.01O nanoparticles obtained from the co-precipitated method. The optical properties show the different optical characteristics of Cd0.99Ni0.01O nanoparticles with 2.06 eV energy gab. The Cd0.99Ni0.01O nanoparticles are employed for some medicinal and pharmaceutical applications. Cytotoxicity of Cd0.99Ni0.01O against human neuroblastoma IMR-32 and cervical cancer HeLa cell lines was studied. The cytotoxicity of Cd0.99Ni0.01O on the previous cancer cell lines resulted in inhibition of cell growth estimated by MTT assay. The exposure of IMR-32 and HeLa cancer cells to Cd0.99Ni0.01O for 48hrs verified a potential apoptotic activity by significant up-regulation of p53 gene expression. Furthermore, anticancer activity was tested by down-regulation of platelet-derived growth factor receptor beta (PDGFR-β) gene expression.

Computational Analysis Reveals the Characteristics of Immune Cells in Glomerular and Tubulointerstitial Compartments in IgA Nephropathy Patients.

Objective: The commonalities and differences regarding immune states between glomerular and tubulointerstitial compartments of IgA nephropathy (IgAN) remains largely undetermined. We aim to perform bioinformatic analysis for providing a comprehensive insight into the characteristics of immune cells and associated molecular mechanisms in IgAN. Materials and Methods: We performed integrated bioinformatic analyses by using IgAN-related datasets from the Gene Expression Omnibus database. First, the differentially expressed genes (DEGs) were identified and subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Then, CIBERSORT was employed to determine the landscape of infiltrating immune cells in both glomerular and tubulointerstitial compartments of IgAN patients, followed by Pearson’s correlation analysis and principal component analysis (PCA). Finally, commonly shared DEGs between glomerular and tubulointerstitial entities were recognized, followed by correlation analyses to identify the dominant commonly shared DEGs associated with immune cell infiltration in IgAN. Results: GO and KEGG enrichment analyses showed apparently distinct biological processes in the glomerular and tubulointerstitial compartments of IgAN. In addition, CIBERSORT analyses revealed a clear trend of increasing proportions of M1 macrophage and M2 macrophage in the glomerular compartment while noticeably higher proportions of resting CD4+ memory T cells and M2 macrophages in the tubulointerstitial compartments. The PCA analyses showed that the varying composition of immune cells in both glomerular and tubulointerstitial entities was compelling to distinguish IgAN patients from healthy living controls. In addition, 21 commonly shared DEGs between glomerular and tubulointerstitial entities were recognized as key regulators in the pathogenesis of IgAN, among which the enhanced hemoglobin subunit beta (HBB) gene expression was found to be positively associated with M2 macrophage in the glomerular compartment and resting CD4+ memory T cells in the tubulointerstitial compartment. Most importantly, FBJ murine osteosarcoma viral oncogene homolog B (FOSB) gene deficiency was recognized as the dominant alteration in promoting M2 macrophage infiltration in the glomerular compartment of IgAN. Conclusion: The findings from our current study for the first time reveal commonalities and differences regarding immune states between glomerular and tubulointerstitial compartments, as well as decode the essential role of M2 macrophages and associated molecular patterns within the microenvironments of IgAN.

Comparison of the Toxicological Effects of Pesticides in Non-Tumorigenic MCF-12A and Tumorigenic MCF-7 Human Breast Cells.

Humans are exposed to residues of organophosphate and neonicotinoid pesticides, commonly used in agriculture. Children are particularly vulnerable and, among possible adverse outcomes, the increased incidence of premature mammary gland development (thelarche) has raised concern. We evaluated the toxicological effects of chlorpyrifos (CPF), imidacloprid (IMI) and glyphosate (GLY) at exposure concentrations occurring in children on the tumorigenic MCF-7 and non-tumorigenic MCF-12A breast cell lines, as representative of the target organ model, assessing cytotoxicity, apoptosis, necrosis, intracellular reactive oxygen species (ROS) and ATP levels, 17β-estradiol secretion and gene expression of nuclear receptors involved in mammary gland development. The pesticides decreased cell vitality in MCF-7 and cell proliferation in MCF-12A cells. ATP levels were decreased in MCF-7 cells by pesticides and apoptosis was increased in MCF-12A cells only by GLY (2.3 nM). ROS production was decreased by pesticides in both cell lines, except IMI (1.6 nM) in MCF-7 cells. Endocrine disrupting activity was highlighted by induction of 17β-estradiol secretion and modulation of the gene expression of estrogen alpha and beta, progesterone, androgen, and aryl hydrocarbon receptors in both cell lines. The use of MCF-7 and MCF-12A cells highlighted dissimilar modes of action of each pesticide at low human relevant concentrations.

Inflammasomes in placental explants of women with preeclampsia cultured with monosodium urate may be modulated by vitamin D

ABSTRACT Objectives Preeclampsia (PE) is an important syndrome of gestation characterized by placental and systemic inflammation. High plasma concentration of uric acid are frequently associated with inflammation and endothelial dysfunction and may contribute to PE pathogenesis. This study aimed to evaluate the vitamin D (VD) immunomodulatory effect on the NLRP1/NLRP3 inflammasomes in placental explants from preeclamptic (PE) and normotensive (NT) pregnant women. Study design Placental explants from 10 late-onset PE (LOPE), 10 early-onset PE (EOPE), and 10 NT pregnant women were cultured with or without monosodium urate (MSU) and VD. Main outcome measures Gene and protein expression of NLRP1, NLRP3, HMGB1, caspase-1, interleukin-1 beta (IL-1β), and IL-18 were determined by quantitative PCR and Western blotting/ELISA. Statistical significance was accepted at p < 0.05. Results Basal gene and protein expression of NLRP1/NLRP3 and IL-1β, IL-18 and HMGB1 were significantly higher in explants from EOPE compared to LOPE and NT pregnant women. In addition, culture with MSU increased these inflammatory markers, and concomitant treatment with MSU+VD decreased this effect. Conclusions The results demonstrated that NLRP1 and NLRP3 inflammasomes are upregulated in the placental tissue of EOPE women, associated with high production of inflammatory cytokines. The in vitro treatment with VD downregulated placental inflammasomes induced by MSU, suggesting its immunomodulatory role in the systemic inflammation of PE.

Noradrenaline and ATP regulate adiponectin exocytosis in white adipocytes: Disturbed adrenergic and purinergic signalling in obese and insulin-resistant mice

White adipocyte adiponectin exocytosis is triggered by cAMP and a concomitant increase of cytosolic Ca2+ potentiates its release. White adipose tissue is richly innervated by sympathetic nerves co-releasing noradrenaline (NA) and ATP, which may act on receptors in the adipocyte plasma membrane to increase cAMP via adrenergic receptors and Ca2+ via purinergic receptors. Here we determine the importance of NA and ATP for the regulation of white adipocyte adiponectin exocytosis, at the cellular and molecular level, and we specifically detail the ATP signalling pathway. We demonstrate that tyrosine hydroxylase (enzyme involved in catecholamine synthesis) is dramatically reduced in inguinal white adipose tissue (IWAT) isolated from mice with diet-induced obesity; this is associated with diminished levels of NA in IWAT and with a reduced ratio of high-molecular-weight (HMW) to total adiponectin in serum. Adiponectin exocytosis (measured as an increase in plasma membrane capacitance and as secreted product) is triggered by NA or ATP alone in cultured and primary mouse IWAT adipocytes, and enhanced by a combination of the two secretagogues. The ATP-induced adiponectin exocytosis is largely Ca2+-dependent and activated via purinergic P2Y2 receptors (P2Y2Rs) and the Gq11/PLC pathway. Adiponectin release induced by the nucleotide is abrogated in adipocytes isolated from obese and insulin-resistant mice, and this is associated with ∼70% reduced abundance of P2Y2Rs. The NA-triggered adiponectin exocytosis is likewise abolished in “obese adipocytes”, concomitant with a 50% lower gene expression of beta 3 adrenergic receptors (β3ARs). An increase in intracellular Ca2+ is not required for the NA-stimulated adiponectin secretion. Collectively, our data suggest that sympathetic innervation is a principal regulator of adiponectin exocytosis and that disruptions of this control are associated with the obesity-associated reduction of circulating levels of HMW/total adiponectin.

Clinical and Biological Significance of DNA Methylation-Driven Differentially Expressed Genes in Biochemical Recurrence After Radical Prostatectomy.

Background: Biochemical recurrence (BCR) after radical prostatectomy indicates poor prognosis in patients with prostate cancer (PCA). DNA methylation (DNAm) is a critical factor in tumorigenesis and has attracted attention as a biomarker for the diagnosis, treatment, and prognosis of PCA. However, the predictive value of DNAm-derived differentially expressed genes (DMGs) in PCA with BCR remains elusive. Methods: We filtered the methylated genes and the differentially expressed genes (DGEs) for more than 1,000 clinical samples from the TCGA cohort using the chAMP and DESeq2 packages of R language, respectively. Next, we integrated the DNAm beta value and gene expression data with the Mithymix package of R language to obtain the DMGs. Then, 1,000 times Cox LASSO regression with 10-fold cross validation was performed to screen signature DMGs and establish a predictive classifier. Univariate and multivariate cox regressive analyses were used to identify the prognostic factors to build a predictive model, and its performance was measured by receiver operating characteristic, calibration curves, and Harrell’s concordance index (C-index). Additionally, a GEO dataset was used to validate the prognostic classifier. Results: One hundred DMGs were mined using the chAMP and Methymix packages of R language. Of these, seven DMGs (CCK, CD38, CYP27A1, EID3, HABP2, LRRC4, and LY6G6D) were identified to build the prognostic classifier (Classifier) through LASSO analysis. Moreover, univariate and multivariate Cox regression analysis determined that the Classifier and pathological T stage (pathological_T) were independent predictors of BCR (hazard ratio (HR 2.2), (95% CI 1.4–3.5), p < 0.0012, and (HR 1.8), (95% CI 1.0–3.2), p < 0.046). A nomogram based on the Classifier was constructed, with high prediction accuracy for BCR-free survival in TCGA and GEO datasets. GSEA enrichment analysis showed that the DMGs were mainly enriched in the metabolism pathways. Conclusion: We identified and validated the nomogram of BCR-free survival for PCA patients, which has the potential to guide treatment decisions for patients at differing risks of BCR. Our study deepens the understanding of DMGs in the pathogenesis of PCA.

Association of Elevated Plasma FGF21 and Activated FGF21 Signaling in Visceral White Adipose Tissue and Improved Insulin Sensitivity in Gestational Diabetes Mellitus Subtype: A Case-Control Study.

ObjectiveTo study the discrepancy of the insulin sensitivity alteration pattern, circulating fibroblast growth factor (FGF21) levels and FGF21 signaling in visceral white adipose tissue (vWAT) of gestational diabetes mellitus (GDM) subtypes.Methods26 GDM women with either a predominant of insulin-secretion defect (GDM-dysfunction, n = 9) or insulin-sensitivity defect (GDM-resistance, n = 17) and 13 normal glucose tolerance (NGT) women scheduled for caesarean-section at term were studied. Blood and vWAT samples were collected at delivery.ResultsThe insulin sensitivity was improved from the 2nd trimester to delivery in the GDM-resistance group. Elevated circulating FGF21 concentration at delivery, increased FGF receptor 1c and decreased klotho beta gene expression, enhanced ERK1/2 phosphorylation, and increased GLUT1, IR-B, PPAR-γ gene expression in vWAT were found in the GDM-resistance group as compared with the NGT group. The circulating FGF21 concentration was negatively correlated with fasting blood glucose (r = -0.574, P < 0.001), and associated with the GDM-resistance group (r = 0.574, P < 0.001) in pregnant women at delivery. However, we observed no insulin sensitivity alteration in GDM-dysfunction and NGT groups during pregnancy. No differences of plasma FGF21 level and FGF21 signaling in vWAT at delivery were found between women in the GDM-dysfunction and the NGT group.ConclusionsWomen with GDM heterogeneity exhibited different insulin sensitivity alteration patterns. The improvement of insulin sensitivity may relate to the elevated circulating FGF21 concentration and activated FGF21 signaling in vWAT at delivery in the GDM-resistance group.

Glucokinase activation leads to an unsustained hypoglycaemic effect with hepatic triglyceride accumulation in db/db mice.

To investigate how subchronic administration of a glucokinase activator (GKA) results in attenuation of the hypoglycaemic effect in the diabetic condition. Six-week-old db/db mice were fed standard chow containing a GKA or the sodium-glucose cotransporter 2 inhibitor ipragliflozin for 1, 6, 14 or 28 days. We performed histological evaluation and gene expression analysis of the pancreatic islets and liver after each treatment and compared the results to those in untreated mice. The unsustained hypoglycaemic effect of GKAs was reproduced in db/db mice in conjunction with significant hepatic fat accumulation. The initial reactions to treatment with the GKA in the liver were upregulation of the gene expression of carbohydrate response element-binding protein beta (Chrebp-b) and downregulation of phosphoenolpyruvate carboxykinase (Pepck) on day 1. Subsequently, the initial changes in Chrebp-b and Pepck disappeared and increases in the expression of genes involved in lipogenesis, including acetyl-CoA carboxylase and fatty acid synthase, were observed. There were no significant changes in the pancreatic β cells nor in hepatic insulin signalling. The GKA showed an unsustained hypoglycaemic effect and promoted hepatic fat accumulation in db/db mice. Dynamic changes in the expression of hepatic genes involved in lipogenesis and gluconeogenesis could affect the unsustained hypoglycaemic effect of the GKA despite no changes in pancreatic β-cell function and mass.

Use of paclitaxel carried in lipid nanoparticles to treat aortic allograft transplantation in rats.

The aim of this study was to test whether lipid core nanoparticles loaded with paclitaxel (LDE-PTX) protect rat aortic allograft from immunological damage. Fisher and Lewis rats were used differing in minor histocompatibility loci. Sixteen Lewis rats were allocated to four-animal groups: SYNG (syngeneic), Lewis rats receiving aorta grafts from Lewis rats; ALLO (allogeneic), Lewis rats receiving aortas from Fisher rats; ALLO+LDE (allogeneic transplant treated with LDE), Lewis rats receiving aortas from Fisher rats, treated with LDE (weekly injection for 3 weeks); ALLO+LDE-PTX (allogeneic transplant treated with LDE-PTX), Lewis rats receiving aortas from Fisher rats treated with LDE-PTX (4 mg/kg weekly for 3 weeks). Treatments began on transplantation day. Thirty days post-transplantation, SYNG showed intact aortas. ALLO and ALLO+LDE presented intense neointimal formation. In ALLO+LDE-PTX, treatment inhibited neointimal formation; narrowing of aortic lumen was prevented in ALLO and ALLO+LDE. LDE-PTX strongly inhibited proliferation and intimal invasion by smooth muscle cells, diminished 4-fold presence of apoptotic/dead cells in the intima, reduced the invasion of aorta by macrophages and T-cells and gene expression of pro-inflammatory tumour necrosis factor-alpha (TNFα), interferon gamma (IFNγ) and interleukin-1 beta (IL-1β). LDE-PTX was effective in preventing the vasculopathy associated with rejection and may offer a potent therapeutic tool for post-transplantation.

Experimental models of orthodontic tooth movement and their effects on periodontal tissues remodelling

ObjectivesThe present study aimed to compare two different models of orthodontic tooth movement (OTM) in rats by evaluating tooth movement efficiency and periodontal tissues remodelling. DesignFifteen animals were randomly distributed into 3 groups: control group (untreated); ligature appliance (LA) as experimental OTM using a closed coil spring fixed around maxillary first molar by steel ligature; occlusal appliance (OA) as experimental OTM using a closed coil spring attached on the occlusal surface of the maxillary first molar. After 15 days, all animals were euthanized, and the maxilla of each animal was collected for qPCR, micro-computed tomography, and histological analyses. ResultsInterleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha gene expressions were significantly upregulated in the animals of the LA group as compared to the other groups. No significant difference was observed in tooth displacement between both methods. The LA group presented higher linear bone loss and lower values of bone volume fraction, bone mineral density, trabecular number and increased values of trabecular separation compared to the other groups. The birefringent collagen content in the tension side of the periodontal ligament contained significantly lower collagen content in the LA group than in the control group. Furthermore, on the pressure side, the collagen content was significantly lower in the LA and OA groups than in the control group. ConclusionsThe OA group presented little or no deleterious effect on periodontal tissues compared to the LA group, suggesting its use may be more reliable for OTM induction in rats for 15 days.

Serum Level Determination of Some Immunological Parameters and STAT Genes in Pediatrics SLE Patients Compared to Healthy Control

Systemic lupus erythematosus was defined as an autoimmune chronic systemic rheumatic disease. it has many disease phenotypes and clinical symptoms varying from one patient to another, which include mild mucocutaneous manifestations to severe involvement of the central nervous system and various organs in the body. Several pathogenic pathways of immunity play a part in the progression of SLE. The aims of this study are to determine the serum levels of C3, C4, IL-9, IL-10, IL-12 and INF-gamma and disease activity in pediatric Iraqi (pSLE) patients additionally in this study we aim to find the levels of STAT-alpha and beta gene expression change in patients.30 samples were collected from pediatric SLE patients and 30 healthy samples. The serum levels of all the immunological parameters were evaluated by ELISA and the gene expression of STAT-alpha and beta were evaluated by RT-PCR. The results showed lower level in both complements components while increased levels in all interleukins. The gene expression showed increased levels of both genes.

Dasatinib-Induced Lymphocyte Elevation and Imbalance of T Cell Receptor V Beta Gene Expression in Chronic Phase Chronic Myelogenous Leukemia Patients

Introduction: Chronic myelogenous leukemia (CML) is a myeloproliferative neoplasms caused by a reciprocal translocation of chromosomes 9 and 22 [t(9;22)], producing the BCR-ABL fusion gene. Imatinib (Ima), the 1st generation tyrosine kinase inhibitor (TKI), has drastically improved the prognosis of CML compared to previous therapies. Imatinib and the 2nd-generation TKIs dasatinib (Dasa) and nilotinib (Nilo) are generally used for chronic phase CML (CP-CML) as induction chemotherapy. Lymphocyte proliferation and increased large granular lymphocytes (LGLs) are sometimes observed in patients treated with Dasa. These LGLs have been reported to be mainly natural killer (NK) cells. However, there is insufficient data about T-lymphocytes, including CD8 T cells, in CML patients treated with TKIs. We analyzed lymphocyte counts and percentages in CP-CML patients treated with Dasa and other TKIs. We also examined T cell receptor (TCR) V beta gene repertoire in some patients.Methods: The patients in this study were CP-CML patients diagnosed at our hospital from August 2006 to June 2016 and treated with TKIs. The main aims were to examine (1) lymphocyte rates and (2) cytotoxic T lymphocytes (CTLs) and TCR V beta gene repertoire. These analyses were conducted using flow cytometry. All statistical analyses were performed using GraphPad Prism 6 software.Results: Table 1 summarizes data comparing patients treated with Dasa and those treated with other TKIs. Six patients were treated with Dasa only. Seven and 2 patients, respectively, were treated with Ima or Nilo only. Eleven patients were treated with multiple TKIs including Dasa, and 2 patients were treated with multiple TKIs not including Dasa. The median age at first TKI administration was 60 years in patients treated with Dasa and 72 years in patients treated with other TKIs. No significant statistical difference was observed in age at first TKI administration. The median follow-up period of patients treated with TKIs without Dasa was significantly longer than that of patients treated with TKIs including Dasa. This was due to the difference in the timing of approval of Ima (1st generation TKI) and Dasa (2nd generation TKI). There was no significant difference in the best response between the Dasa and non-Dasa groups. There was also no significant difference in overall survival between the 2 groups (Log-rank, P = 0.7283). Pleural effusion was observed in 5 patients in the Dasa group and 1 in the non-Dasa group, without statistical significance. Figure 1 shows lymphocyte elevation in the Dasa group compared to the non-Dasa group with regard to both lymphocyte counts and percentages. Figure 2 shows the time courses of lymphocyte percentages. Counts from 1, 3, 6 and 12 months from the initiation of Dasa (A) and non-Dasa TKIs (B) are indicated. The lymphocyte rates in the non-Dasa group showed no significant change (Figure 2B). However, gradual lymphocyte elevation was clearly seen with the passage of time in Dasa group patients, with statistical significance (Figure 2A). The time courses of lymphocyte counts in 2 cases are shown in Figure 3. Lymphocyte elevations were clearly observed after changing to Dasa from Nilo (Figure 3A) and Ima (Figure 3B). We also analyzed the detailed lymphocyte fraction in some patients. Representative data of a patient treated with Dasa only are shown in Table 2 and Figure 4. NK cells were found in the peripheral blood, as have been seen in previous reports, and, interestingly, CTLs were also detected at all time points (Table 2). It is notable that about 20% of the peripheral blood consisted of CTLs 35 months after the first administration, which was the same percentage as that of NK cells. TCR V beta gene repertoire in the same patient is shown in Figure 4. Oligoclonal CD8 T cell expansion was observed using V beta 13.1, V beta 7.1, and V beta 2 expression. These results suggest that not only NK cells but also CTLs may play an important role in attacking CML blasts in patients treated with Dasa.Conclusions: In CP-CML patients treated with TKIs, lymphocyte elevation was obviously observed only in patients treated with Dasa. CTL analyses and TCR V beta gene analyses suggest that CTLs can be induced along with NK cells in patients treated with Dasa. These results show that Dasa can work in a different way than other TKIs. Clarifying the tumor antigens recognized by these CTLs may bring new insights and connect to new cellular therapies for refractory CML cases. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Correlations between Low Doses of Zearalenone, Its Carryover Factor and Estrogen Receptor Expression in Different Segments of the Intestines in Pre-Pubertal Gilts-A Study Protocol.

Plant materials can be contaminated with Fusarium mycotoxins and their derivatives, whose toxic effects on humans and animals may remain subclinical. Zearalenone (ZEN), a low-molecular-weight compound, is produced by molds in crop plants as a secondary metabolite. The objective of this study will be to analyze the in vivo correlations between very low monotonic doses of ZEN (5, 10, and 15 μg ZEN/kg body weight—BW for 42 days) and the carryover of this mycotoxin and its selected metabolites from the intestinal contents to the intestinal walls, the mRNA expression of estrogen receptor alfa (ERα) and estrogen receptor beta (ERβ) genes, and the mRNA expression of genes modulating selected colon enzymes (CYP1A1 and GSTP1) in the intestinal mucosa of pre-pubertal gilts. An in vivo experiment will be performed on 60 clinically healthy animals with initial BW of 14.5 ± 2 kg. The gilts will be randomly divided into a control group (group C, n = 15) and three experimental groups (group ZEN5, group ZEN10, and group ZEN15; n = 15). Group ZEN5 will be administered per os 5 μg ZEN/kg BW (MABEL), group ZEN10—10 μg ZEN/kg BW (NOAEL), and group ZEN15—15 µg ZEN/kg BW (low LOAEL). In each group, five animals will be euthanized on analytical dates 1 (exposure day 7), 2 (exposure day 21), and 3 (exposure day 42). Samples for in vitro analyses will be collected from an intestinal segment resected from the following regions: the third (horizontal) part of the duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, and descending colon. The experimental material will be collected under special conditions, and it will be transported to specialist laboratories where samples will be obtained for further analyses.

Effect of anthocyanin extract from Roselle,Hibiscus sabdariffa, calyx on haematological, biochemical and immunological parameters of rainbow trout,Oncorhynchus mykiss

In the present study, rainbow trout, Oncorhynchus mykiss, (14.1 ± 0.08 g) were fed diets supplemented with different levels of Roselle anthocyanin extract (RE) to assess the fish immune, stress and haematological parameters. For this, four levels of RE including 0, 0.5, 1 and 1.5% were added to the fish diet for eight weeks. Then, growth performance, blood haematological, biochemical, immunological and hepatic expression of cytokine genes were investigated in all treatments. At the end of the experiment, the fish exhibited similar growth rate and survival (final weight: 91.4–96.7 g; feed conversion ratio: 0.75–0.80; specific growth rate: 3.11–3.21% d−1; survival: 100%). The results showed that there were no significant differences in plasma glucose, cholesterol, triglyceride, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and alkaline phosphatase levels among the treatments. Supplementation of the diets with 1% RE significantly increased RBC, haemoglobin, haematocrit, lysozyme, complement and WBC, compared with the control treatment. 1 and 1.5% RE significantly decreased plasma cortisol levels, compared with the control. All levels of dietary RE significantly up-regulated hepatic tumour necrosis factor-alpha gene expression, compared with the control treatment. 1 and 1.5% RE significantly up-regulated hepatic interleukin-1 beta gene expression, compared with the control treatment. 1.5% RE supplementation significantly down-regulated interleukin-10 gene expression, compared with the other treatments. In conclusion, dietary RE supplementation at 1% is beneficial for rainbow trout, as it mitigates stress and increases immune-related parameters.

Enhanced structure and function of human pluripotent stem cell-derived beta-cells cultured on extracellular matrix.

The differentiation of human stem cells into insulin secreting beta‐like cells holds great promise to treat diabetes. Current protocols drive stem cells through stages of directed differentiation and maturation and produce cells that secrete insulin in response to glucose. Further refinements are now needed to faithfully phenocopy the responses of normal beta cells. A critical factor in normal beta cell behavior is the islet microenvironment which plays a central role in beta cell survival, proliferation, gene expression and secretion. One important influence on native cell responses is the capillary basement membrane. In adult islets, each beta cell makes a point of contact with basement membrane protein secreted by vascular endothelial cells resulting in structural and functional polarization. Interaction with basement membrane proteins triggers local activation of focal adhesions, cell orientation, and targeting of insulin secretion. This study aims to identifying the role of basement membrane proteins on the structure and function of human embryonic stem cell and induced pluripotent stem cell‐derived beta cells. Here, we show that differentiated human stem cells‐derived spheroids do contain basement membrane proteins as a diffuse web‐like structure. However, the beta‐like cells within the spheroid do not polarize in response to this basement membrane. We demonstrate that 2D culture of the differentiated beta cells on to basement membrane proteins enforces cell polarity and favorably alters glucose dependent insulin secretion.

Influence of prenatal stress on metabolic abnormalities induced by postnatal intake of a high-fat diet in BALB/c mice.

Prenatal insults during fetal development result in increased likelihood of developing chronic disease. Obesity, the biggest risk factor for the development of metabolic disease, is affected by several genetic and environmental factors. High-fat diet (HFD) consumption is usually linked with the development of obesity. The main goal of this study was to analyze the impact of the exposure to a HFD in prenatally stressed animals. For this purpose, we subjected pregnant BALB/c mice to restraint stress for 2 h a day between gestational day (GD) 14 and GD 21. Prenatally stressed and control offspring of both sexes were postnatally exposed to a HFD for 24 weeks. We found that prenatal stress (PS) per se produced disturbances in males such as increased total blood cholesterol and triglycerides, with a decrease in mRNA expression of sirtuin-1. When these animals were fed a HFD, we observed a rise in glucose and insulin levels and an increase in visceral adipose tissue gene expression of leptin, resistin, and interleukin-1 beta. Although females proved to be more resilient to PS consequences, when they were fed a HFD, they showed significant metabolic impairment. In addition to the changes observed in males, females also presented an increase in body weight and adiposity and a rise in cholesterol levels.