scholarly journals Histological study of gonadal tissues of adult Artemia salina (Linnaeus 1758) and immunohistochemistry by Caspase 3 and HSP70 to detect specific apoptosis markers on gonadal tissues after exposure to TBTCl

2021 ◽  
Vol 11 (1) ◽  
pp. 112-120
Author(s):  
Najla Mohamed Abushaala ◽  
Abdulfattah Mohamed Elfituri ◽  
Syaizwan Zahmir Zulkifli
Keyword(s):  
Immune System ◽  
Caspase 3 ◽  
Biological Effects ◽  
Artemia Salina ◽  
Follicle Cells ◽  
Hsp70 Expression ◽  
Gonad Tissue ◽  
Control Tissue ◽  
Tributyltin Chloride ◽  
Apoptosis Markers

Background: Several types of research have been recently carried out on the biological effects of TBTs, including investigations of genitals in invertebrates in response to exposure to TBTs in marine water. Aim: The objective of this research was to investigate the acute effects of tributyltin chloride (TBTCl) on gonads in the adult stage of Artemia salina by use normal histology and immunohistochemistry (IHC) (Caspase 3 and HSP70) to see specific apoptosis markers. Methods: After exposure of A. salina to different concentrations of TBTCl (25, 50, 100, 200, and 300 ng.l−1), 50 adult A. salina (25 male and 25 female) were selected randomly from each concentration to histologically study the gonads. The gonad tissue was sectioned (5 μm) and some slides were stained with hematoxylin and eosin and others were stained with IHC avidin–biotin complex, and were examined under a light microscope. Results: The results showed significant differences (p < 0.05) in histological lesions between different concentrations of TBTCl. The histological lesions in the testis and ovary section were undifferentiated cells, degenerating yolk globules, and follicle cells enveloping the oocyte which was then compared with control tissue, and these effects were found to be increased in females more than in males with the highest concentration of TBTCl. Immunohistochemistry (IHC) showed that positive immunostaining was observed in the testis and ovary as brownish deposits to Caspase 3 and HSP70 antibody after exposure to TBTCl, while the testis and ovary section in control tissue had no immunoreactivity to Caspase 3 and HSP70 antibody; these effects were profoundly increased with the highest concentration of TBTCl in females more than in males. Finally, the histological lesions and IHC (Caspase 3 and HSP70) revealed that the apoptosis and immune system stress of A. salina gonad tissue damage in females were more sensitive to TBTCl toxicity as compared to white males. Conclusion: In general, the present study aimed to observe the effects TBTCl on A. salina gonads by using histological sections and IHC (Caspase 3 and HSP70), which were evaluated for the first time and have been proven to possess an important function in apoptosis marker and immune system stress in Artemia. Finally, the specific mechanisms through which TBTCl affects A. salina Caspase 3 and HSP70 expression need further investigation.

Abstract P336: Endogenous Alpha-1a Adrenergic Receptors Promote Cardiomyocyte Survival Through Suppression Of Rip Kinase-mediated Necroptosis

2021 ◽  
Vol 129 (Suppl_1) ◽  
Author(s):  
Jiandong Zhang ◽  
Peyton Sandroni ◽  
Wei Huang ◽  
Brian C Jensen
Keyword(s):  
Adrenergic Receptors ◽  
Adrenergic Receptor ◽  
Caspase 3 ◽  
Underlying Mechanism ◽  
Hmgb1 Level ◽  
Programmed Necrosis ◽  
Apoptosis Markers ◽  
Kinase Pathway ◽  
Cardiomyocyte Survival ◽  
Serum Hmgb1 Level

Our previous work has demonstrated essential protective roles for the endogenous cardiomyocyte alpha-1A adrenergic receptor (α1A-AR) subtype in mouse models of heart failure. However, the underlying mechanism of this protective phenotype is unclear. To address this gap in knowledge, we bred a mouse line lacking α1A-ARs on cardiomyocytes by crossing αMHC-cre mice with floxed α1A mice (CMKO= cre+ fl/fl, CMWT= cre- fl/fl), and subjected males to permanent LAD ligation. CMKO mice had increased serum HMGB1 level, larger infarcts and higher mortality. We found that RIP1/3-mediated programmed necrosis (necroptosis), but not apoptosis was exaggerated in CMKO mice 3 days after ligation. We then tested whether RIP1 inhibition with Nec-1s could mitigate this injury. Mice were given Nec-1s (1.65 mg/kg) or vehicle 10 mins prior to LAD ligation, followed by daily IV injection. Nec-1s treatment diminished post-ligation RIP1 (0.62±0.02 vs. 0.78±0.23 A.U., p=NS) and RIP3 expression (0.33±0.1 vs. 0.26±0.10 A.U., p=NS) in CMWT and CMKO mice respectively. Serum level of HMGB1 on D3 was markedly reduced in both CMWT (45.1%) and CMKO (61.1 %) after Nec-1s treatment. There was no difference between Nec-1s treated CMWT and CMKO mice (147±53 vs. 174±37 pg/mL, p=NS), indicating that blocking the RIP kinase pathway abrogates the exaggerated cell death in CMKO mice after ligation. Likewise, Nec-1s-treated CMKO mice had similar infarct areas to CMWT controls (16.2±4.5 vs. 19.9±4.6%, p=NS), further confirming that targeting necroptosis abrogates pathological damage. Collectively these Nec-1s data suggest that RIP-mediated necroptosis may account for larger infarcts in CMKO mice. Interestingly, expression of the apoptosis markers c-caspase-3 and PARP was similar between CMWT and CMKO mice, suggesting that the α1A-AR specifically regulates necroptosis. In sum, our data demonstrate that RIP kinase-mediated necroptosis contributes to susceptibility to injury in mice lacking cardiomyocyte α1A-ARs.

BTK Inhibitors Differentially Induce Apoptosis but Similarly Suppress Chemotaxis and Lipid Accumulation in Mantle Cell Lymphoma

2020 ◽  
Author(s):  
Zhuojun Liu ◽  
Jia Liu ◽  
Tainming Zhang ◽  
Lin Li ◽  
Shuo Zhang ◽  
...  
Keyword(s):  
Mantle Cell Lymphoma ◽  
Caspase 3 ◽  
Cell Lymphoma ◽  
Stimulated Raman Scattering ◽  
Biological Effects ◽  
Immunoblot Analysis ◽  
Mantle Cell ◽  
Apoptotic Genes ◽  
Btk Inhibitors ◽  
Cell Chemotaxis

Abstract Background: The more selective second-generation BTK inhibitors (BTKis) Acalabrutinib and Zanubrutinib and the first-generation BTK inhibitor (BTKi) Ibrutinib are highlighted by their clinical effectiveness in mantle cell lymphoma (MCL), however, similarities and differences of their biological and molecular effects on anti-survival of MCL cells induced by these BTKis with distinct binding selectivity against BTK remain largely unknown. Methods: AlamarBlue assays were performed to define cytotoxicity of BTKis against MCL cells, Jeko-1 and Mino. Cleaved PARP and caspase-3 levels were examined by immunoblot analysis to study BTKi-induced apoptotic effects. Biological effects of BTKis on MCL-cell chemotaxis and lipid droplet (LD) accumulation were examined in Jeko-1, Mino and primary MCL cells via Transwell and Stimulated Raman scattering imaging analysis respectively. Enzyme-linked immunoassays were used to determine CCL3 and CCL4 levels in MCL-cell culture supernatants. RNA-seq analyses identified BTKi targets which were validated by quantitative RT-PCR (qRT-PCR) and immunoblot analysis. Results: Acalabrutinib and Zanubrutinib induced moderate apoptosis in Ibrutinib high-sensitive JeKo-1 cells and Ibrutinib low-sensitive Mino cells, which was accompanied by cleaved PARP and caspase-3. Such effects might be caused by the stronger ability of Ibrutinib to upregulate the expression of pro-apoptotic genes, such as HRK, GADD45A , and ATM , in JeKo-1 cells than in Mino cells, and the expression of such apoptotic genes was slightly changed by Acalabrutinib and Zanubrutinib in both JeKo-1 and Mino cells. Further, Acalabrutinib, Zanubrutinib and Ibrutinib reduced MCL-cell chemotaxis with similar efficiency, due to their similar abilities to downmodulate chemokines, such as CCL3 and CCL4. Also, these three BTKis similarly suppressed MCL-cell LD accumulation via downregulating lipogenic factors, DGAT2, SCD, ENPP2 and ACACA without significant differences. Conclusion: BTKis demonstrated differential capacities to induce MCL-cell apoptosis due to their distinct capabilities to regulate the expression of apoptosis-related genes, and similar biological and molecular inhibitory effects on MCL-cell chemotaxis and LD accumulation.

scholarly journals Comparison of Apoptosis Detection Markers Combined with Macrophage Immunostaining to Study Phagocytosis of Apoptotic Cells in Situ

2006 ◽  
Vol 1 ◽  
pp. 117727190600100 ◽  
Author(s):  
Dorien M. Schrijvers ◽  
Guido R.Y. De Meyer ◽  
Mark M. Kockx ◽  
Arnold G. Herman ◽  
Wim Martinet
Keyword(s):  
Lupus Erythematosus ◽  
Caspase 3 ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Apoptotic Cells ◽  
Immunological Responses ◽  
Apoptosis Detection ◽  
Apoptosis Markers ◽  
Suitable Marker ◽  
Parp 1

Efficient phagocytosis of cells undergoing apoptosis by macrophages is important to prevent immunological responses and development of chronic inflammatory disorders such as systemic lupus erythematosus, cystic fibrosis and atherosclerosis. To study phagocytosis of apoptotic cells (AC) by macrophages in tissue, we validated different apoptosis markers (DNA fragmentation, caspase-3 activation and cleavage of its substrate poly(ADP-ribose)polymerase-1) in combination with macrophage immunostaining. Human tonsils were used as a model because they show a high apoptosis frequency under physiological conditions as well as efficient phagocytosis of AC by macrophages. On the other hand, advanced human atherosclerotic plaques were examined since plaques show severely impaired phagocytosis of AC. Our results demonstrate that the presence of non-phagocytized terminal deoxynucleotidyl transferase end labelling (TUNEL)-positive AC represents a suitable marker of poor phagocytosis by macrophages in situ. Other markers for apoptosis, such as cleavage of caspase-3 or PARP-1, should not be used to assess phagocytosis efficiency, because activation of the caspase cascade and cleavage of their substrates can occur in AC when they have not yet been phagocytized by macrophages.

scholarly journals Uric acid and inflammation in kidney disease

2020 ◽  
Vol 318 (6) ◽  
pp. F1327-F1340 ◽  
Author(s):  
Su Woong Jung ◽  
Su-Mi Kim ◽  
Yang Gyun Kim ◽  
Sang-Ho Lee ◽  
Ju-Young Moon
Keyword(s):  
Uric Acid ◽  
Immune System ◽  
Kidney Disease ◽  
Biological Effects ◽  
Experimental Studies ◽  
Epidemiologic Studies ◽  
Causative Role ◽  
Immune System Activation ◽  
Progression Of Kidney Disease ◽  
System Activation

Asymptomatic hyperuricemia is frequently observed in patients with kidney disease. Although a substantial number of epidemiologic studies have suggested that an elevated uric acid level plays a causative role in the development and progression of kidney disease, whether hyperuricemia is simply a result of decreased renal excretion of uric acid or is a contributor to kidney disease remains a matter of debate. Over the last two decades, multiple experimental studies have expanded the knowledge of the biological effects of uric acid beyond its role in gout. In particular, uric acid induces immune system activation and alters the characteristics of resident kidney cells, such as tubular epithelial cells, endothelial cells, and vascular smooth muscle cells, toward a proinflammatory and profibrotic state. These findings have led to an increased awareness of uric acid as a potential and modifiable risk factor in kidney disease. Here, we discuss the effects of uric acid on the immune system and subsequently review the effects of uric acid on the kidneys mainly in the context of inflammation.

scholarly journals Nodal induces apoptosis through activation of the ALK7 signaling pathway in pancreatic INS-1 β-cells

2012 ◽  
Vol 303 (1) ◽  
pp. E132-E143 ◽  
Author(s):  
Fang Zhao ◽  
Fengjie Huang ◽  
Mengxiong Tang ◽  
Xiaoming Li ◽  
Nina Zhang ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
Biological Effects ◽  
Cell Mass ◽  
Type I ◽  
Β Cells ◽  
Β Cell ◽  
Akt Phosphorylation ◽  
Induced Apoptosis

We demonstrated previously that the activation of ALK7 (activin receptor-like kinase-7), a member of the type I receptor serine/threonine kinases of the TGF-β superfamily, resulted in increased apoptosis and reduced proliferation through suppression of Akt signaling and the activation of Smad2-dependent signaling pathway in pancreatic β-cells. Here, we show that Nodal activates ALK7 signaling and regulates β-cell apoptosis. We detected Nodal expression in the clonal β-cell lines and rodent islet β-cells. Induction of β-cell apoptosis by treatment with high glucose, palmitate, or cytokines significantly increased Nodal expression in clonal INS-1 β-cells and isolated rat islets. The stimuli induced upregulation of Nodal expression levels were associated with elevation of ALK7 protein and enhanced phosphorylated Smad3 protein. Nodal treatment or overexpression of Nodal dose- or time-dependently increased active caspase-3 levels in INS-1 cells. Nodal-induced apoptosis was associated with decreased Akt phosphorylation and reduced expression level of X-linked inhibitor of apoptosis (XIAP). Remarkably, overexpression of XIAP or constitutively active Akt, or ablation of Smad2/3 activity partially blocked Nodal-induced apoptosis. Furthermore, siRNA-mediated ALK7 knockdown significantly attenuated Nodal-induced apoptosis of INS-1 cells. We suggest that Nodal-induced apoptosis in β-cells is mediated through ALK7 signaling involving the activation of Smad2/3-caspase-3 and the suppression of Akt and XIAP pathways and that Nodal may exert its biological effects on the modulation of β-cell survival and β-cell mass in an autocrine fashion.

Effect of Exposure to RF of Fertilized Chicken Eggs and Pregnant Mice on Hatchability, Organ-Weight, and Locally Delayed Hypersensitivity

1993 ◽  
Vol 5 (3) ◽  
pp. 244-247
Author(s):  
Kenichi Saito ◽  
◽  
Yukari Tsuchida ◽  
Kouichiro Yamada ◽  
Masahiro Sugiyamaand Nobuo Goto ◽  
...  
Keyword(s):  
Immune System ◽  
Power Density ◽  
Chronic Exposure ◽  
Biological Effects ◽  
Delayed Hypersensitivity ◽  
Incident Power ◽  
Prolonged Incubation ◽  
Rf Exposure ◽  
Incident Power Density ◽  
Pregnant Mice

The present study consists of two experiments. The first experiment investigates the influence of chronic exposure of 428MHz radio frequency (RF) with an incident power density of 4mW/cm² on the development of chick embryos. Prolonged incubation was found in exposured eggs as compared to the non-exposured (22 vs 21 days). The average thymus weight in both sexes was smaller than that of the control. RF exposure also led to a significant decrease in the thymic cell density of female chicks. The second experiment was conducted in order to reveal the effects of chronic exposure of the immune system of mice exposed to 428MHz-RF with an incident power density of 1mW/cm² during pregnancy. It was found that RF exposure alters the immune system of mice. RF suppressed the cell-mediated immune-competence by local delayed hypersensitivity. These results suggest that chronic exposure of 428MHz-RF radiation induce biological effects on chick embryo and mice.

scholarly journals Bioactive Potential of Two Marine Picocyanobacteria Belonging to Cyanobium and Synechococcus Genera

2021 ◽  
Vol 9 (10) ◽  
pp. 2048
Author(s):  
Patrizia Pagliara ◽  
Giuseppe Egidio De Benedetto ◽  
Matteo Francavilla ◽  
Amilcare Barca ◽  
Carmela Caroppo
Keyword(s):  
Bioactive Compounds ◽  
Human Cancer ◽  
Biological Effects ◽  
Artemia Salina ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Aqueous Extracts ◽  
Anticancer Activities ◽  
Spectrometry Analysis ◽  
Methanolic Extracts

Coccoid cyanobacteria produce a great variety of secondary metabolites, which may have useful properties, such as antibacterial, antiviral, anticoagulant or anticancer activities. These cyanobacterial metabolites have high ecological significance, and they could be considered responsible for the widespread occurrence of these microorganisms. Considering the great benefit derived from the identification of competent cyanobacteria for the extraction of bioactive compounds, two strains of picocyanobacteria (coccoid cyanobacteria < 3 µm) (Cyanobium sp. ITAC108 and Synechococcus sp. ITAC107) isolated from the Mediterranean sponge Petrosia ficiformis were analyzed. The biological effects of organic and aqueous extracts from these picocyanobacteria toward the nauplii of Artemia salina, sea urchin embryos and human cancer lines (HeLa cells) were evaluated. Methanolic and aqueous extracts from the two strains strongly inhibited larval development; on the contrary, in ethyl acetate and hexane extracts, the percentage of anomalous embryos was low. Moreover, all the extracts of the two strains inhibited HeLa cell proliferation, but methanol extracts exerted the highest activity. Gas chromatography–mass spectrometry analysis evidenced for the first time the presence of β-N-methylamino-l-alanine and microcystin in these picocyanobacteria. The strong cytotoxic activity observed for aqueous and methanolic extracts of these two cyanobacteria laid the foundation for the production of bioactive compounds of pharmacological interest.

172 Expression of proliferation and apoptosis markers in cumulus cells surrounding matured and aged oocytes exposed to luteotropic factors during the second phase of in vitro maturation

2019 ◽  
Vol 31 (1) ◽  
pp. 210
Author(s):  
I. Lebedeva ◽  
O. Mityashova ◽  
A. Smekalova ◽  
E. Montvila ◽  
G. Singina ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Caspase 3 ◽  
Cumulus Cells ◽  
Treated Group ◽  
Second Phase ◽  
Proliferation And Apoptosis ◽  
Oocyte Aging ◽  
Apoptosis Markers

The quality and developmental capacity of mammalian oocytes depends on cooperation with surrounding cumulus cells. The functional state and activity of cumulus cells changes with oocyte maturation, especially during the oocyte transition from metaphase I (MI) to metaphase II (MII) stage. In the present work, effects of 3 luteotropic factors, progesterone (P4), prolactin (PRL), and LH, during the second phase of in vitro maturation (IVM) on the subsequent expression of proliferation and apoptosis markers in bovine cumulus cells surrounding matured and aged oocytes were studied. A total of 1532 cumulus-oocyte complexes (COC) were cultured for 12h in TCM-199 containing 10% fetal calf serum (FCS), 10μg mL−1 porcine FSH, and 10μg mL−1 ovine LH at 38.5°C and 5% CO2. Thereafter, COC were transferred to the following IVM systems: (1) TCM-199 containing 10% FCS (Control 1) and (2) a monolayer of granulosa cells (GC) precultured for 12h in TCM-199 containing 10% FCS (Control 2). In both systems, the medium of experimental groups was supplemented with either P4 (50 ng mL−1) or bovine PRL (50ng mL−1) or ovine LH (5μg mL−1); then, the COC were matured for next 12h. Half of the COC matured for 12h in both systems were cultured for an additional 24h in fresh TCM-199 containing 10% FCS to test long-term hormonal effects during oocyte aging. After culture, the cumulus expression of the proliferation marker proliferating cell nuclear antigen (PCNA) and the pro-apoptotic markers caspase-3 and Bax was assessed by the immunocytochemical method. The data from 4 to 5 replicates using 84 to 106 COC per treatment were analysed by ANOVA. After IVM in System 1, the rate of PCNA-positive cumulus cells was higher (P&lt;0.05) in the PRL-treated group (41.3±1.6%) than in the control (34.6±2.3%) or LH-treated group (29.9±2.9%), but did not differ from that in the P4-treated group (38.2±4.8%). In the presence of GC (System 2), the respective rates did not change but were more variable. Aging of COC matured in both systems led to a 1.4- to 1.9-fold reduction in the proportion of the cells containing the proliferation marker PCNA (P&lt;0.05). Meanwhile, none of the hormones tested had any long-term effect on the proliferative activity of senescent cumulus cells. The rate of cumulus cells expressing caspase-3 in different groups varied from 48.5±4.9 to 53.8±5.8% and did not depend on the hormones, IVM system, or oocyte aging. The proportion of the Bax-positive cells was also unaffected by luteotropic factors but increased 1.4 to 1.6 times (P&lt;0.01) following 24h of COC aging. Our findings indicate that PRL can exert a short-term stimulatory action on the proliferative activity of bovine cumulus cells in the course of the second phase of IVM. Meanwhile, the cumulus expression of pro-apoptotic markers caspase-3 and Bax is not responsive to P4, PRL, or LH during the second step of IVM. The study was supported by the Russian Science Foundation (project 16-16-10069).

Immunostimulatory Potential of Natural Compounds and Extracts: A Review

2020 ◽  
Vol 16 (4) ◽  
pp. 444-454
Author(s):  
Andreea C. Stroe ◽  
Simona Oancea
Keyword(s):  
Immune System ◽  
Plant Species ◽  
Biological Effects ◽  
Scientific Evidence ◽  
Natural Compounds ◽  
Food Supplements ◽  
Human Immune System ◽  
Species Type ◽  
Human Immune ◽  
Immunomodulatory Properties

The proper functioning of human immune system is essential for organism survival against infectious, toxic and oncogenic agents. The present study aimed to describe the scientific evidence regarding the immunomodulatory properties of the main micronutrients and specific phytochemicals. Plants of food interest have the ability to dynamically affect the immune system through particular molecules. Plant species, type of compounds and biological effects were herein reviewed mainly focusing on plants which are not commonly used in food supplements. Several efficient phytoproducts showed significant advantages compared to synthetic immunomodulators, being good candidates for the development of immunotherapeutic drugs.

scholarly journals Effect of Hyperbaric Oxygen on Proliferation and Gene Expression of Human Chondrocytes: AnIn VitroStudy

Cartilage ◽  
2018 ◽  
Vol 10 (4) ◽  
pp. 459-466
Author(s):  
Carolin Melcher ◽  
Birte Sievers ◽  
Nadine Höchsmann ◽  
Frank Düren ◽  
Volkmar Jansson ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hyperbaric Oxygen ◽  
Caspase 3 ◽  
Expression Patterns ◽  
Molecular Testing ◽  
Cell Counts ◽  
Specific Proteins ◽  
Apoptosis Markers ◽  
Collagen Ii

PurposeThe present study investigated the effects of hyperbaric oxygen (HBO) on human chondrocyte proliferation and gene expression patterns.MethodsChondrocyte cultures were transferred to a HBO chamber and exposed to 100% oxygen for 7 consecutive days. Within groups, pressure was varied between 1 and 2 atm and duration of HBO administration was varied among 60, 90, and 120 minutes. Cell counts were performed using the WST-1 assay at 1, 3, 5, and 7 days after initiation of HBO treatment to obtain data to plot a growth curve. Gene expression of apoptosis markers PARP and caspase 3, as well as cartilage specific proteins collagen II and COMP, were detected by reverse transcription polymerase chain reaction.ResultsThe experiments showed that in vitro administration of HBO inhibit chondrocyte growth. When applied compression was increased up to 2 atm, chondrocyte cell count was reduced by half at days 3 and 7 in association with an upregulation of the apoptosis markers PARP and caspase 3 as well as the cartilage specific proteins collagen II and COMP. No significant differences were monitored from varied duration of daily treatment.ConclusionChondrocyte growth was inhibited in vitro by treatment of HBO. This inhibitory effect was even increased by elevating the applied pressure, while molecular testing showed reduced chondrocyte growth. Higher levels of HBO inhibited cell growth even more, but up-regulation of apoptosis specific markers and cartilage specific proteins were seen during administration of high oxygen levels. Thus, it has to be evaluated that there is a critical level of hypo-/hyperoxia required to stimulate or at least maintain chondrocyte cell proliferation.

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