Concentration Of Caspase Research Articles

Evaluation of apoptosis markers in patients with atopic dermatitis

The purpose of the study was to analyze indicators of apoptosis in patients with atopic dermatitis (AD). Patients (N = 88) aged from 12 months to 3 years with varying degrees of disease severity were involved into trial. SCORAD indices were: 68.14±2.63 – in patients with severe AD; 32.03±1.43 – with moderate; and 12.12±1.43 – with mild AD, respectively. Determination of apoptosis markers (Caspase 8, Caspase 9, sCD153, sFas-L, Annexin 5) in blood serum was performed using an enzyme-linked immunosorbent assay (ELISA) using commercial kits from Bender MedSystems® (Austria). A study of apoptosis markers in the blood serum of children with AD revealed a significant decrease (p 0.05) in the concentration of Annexin 5 compared to standard values (by 20 times). A statistically significant decrease (p 0.05) in the concentration levels of Caspase 8 and Caspase 9 in the serum of AD patients was revealed. Thus, the concentrations of Caspase 8 were reduced on by 3 times, and Caspase 9 by 8 times compared to the norm. Also In addition, when determining sFas-L indicators, a statistically significant (p 0.05) decrease in concentration was noted compared to standard indicators. The content of sCD153 in the blood serum in AD patients was significantly higher (p 0.05) than the normative values by 5 times (not detected normally). The most pronounced disturbances in apoptosis indicators were observed in patients with severe atopic dermatitis, (p 0.05), highlighting the importance of studying apoptosis in the context of AD and the possible link between disruptions in this process and the severity of clinical manifestations in patients. The study demonstrates the peculiarities of apoptosis processes in children with AD. On the one hand, these patients have significantly increased activation of signaling systems, on the other hand, the elimination of altered immunocompetent cells does not occur properly, which contributes to the progression and chronicity of immune inflammation in the skin.

Влияние терапии цервикальных неоплазий на иммунологические и апоптотические показатели

Aim: Evaluation of immune response factors in the treatment of mild cervical neoplasia against the background of human papillomavirus in patients of childbearing age. Design: Prospective study. Materials and methods. The study included a survey of 210 patients of fertile age. Among them were 150 patients with human papillomavirus (HPV) and cervical intraepithelial neoplasia, established by histological examination. Patients underwent Pap test, extended colposcopy, determination of HPV type and viral load. Patients with suspected pathology of the cervical zone underwent multifocal biopsy of the cervix. When analyzing changes in the cervix zone, the levels of interferon-γ, interleukin-18, caspase 3 and 9 were evaluated. Results. Among patients with cervical neoplasia of the first degree, the concentration of caspase 3, caspase 9 and interferon-γ at the first stage of the examination was higher (p < 0.05) than the values in the control group (among conditionally healthy women without viral load), while interleukin-18 was significantly lower (p < 0.05). In subgroups I and II, after 3 and 12 months, the concentrations of the studied parameters significantly differed from those in the control group and values before treatment (p < 0.05), while, after 12 months, the values of caspase 3 and caspase 9 in the subgroup where therapy was carried out (subgroup II) were significantly lower than in subgroup I. Also, among the patients, there was a statistical difference in the values of the cytokine series, however, the level of interleukin-18 in subgroup II after 10 days and 12 months was significantly higher than in subgroup I. fluctuations in indicators made it possible to justify the use of immunocorrective therapy for the treatment of a mild neoplastic process of the cervical zone. Conclusion. The results of the study allow optimizing approaches to the treatment of women of childbearing age with mild cervical neoplasia on the background of HPV. Keywords: genotyping, caspase-3, caspase-9, apoptosis, cervical neoplasia, polymerase reaction, interleukin-18, tumor necrosis factor-α, viral load.

Administration of 4-Hydroxy-3,5-Di-Tertbutyl Cinnamic Acid Restores Mitochondrial Function in Rabbits with Cerebral Ischemia

Abstract The aim of the study is to evaluate the effect of 4-hydroxy-3,5- di-tertbutyl cinnamic acid on the change in mitochondrial function under conditions of experimental cerebral ischemia in rabbits. The study was performed on 48 male rabbits, which were used for modeling permanent cerebral ischemia by occlusion of the common carotid arteries. The test compound was administered before modeling ischemia for 14 days and after the occurrence of reproducing ischemia, in a similar time interval. After that, neurological deficit and the parameters of mitochondrial respiration, the intensity of anaerobic processes, the latent opening time of the mitochondrial permeability transition pore, the value of the mitochondrial membrane potential and the concentration of caspase – 3 were determined. The administration of 100 mg/kg of 4-hydroxy-3,5-di-tertbutyl cinnamic acid into the animals reduced neurological deficit and restored the mitochondrial membrane potential. Prophylactic administration of 4-hydroxy- 3,5-di-tertbutyl cinnamic acid, contributed to an increase in ATPgenerating ability, the maximum level of respiration and respiratory capacity by 4.1 times (p<0.01), 4.8 times (p<0.01) and 4.3 times (p<0.01), respectively. With therapeutic administration, these indicators increased by 11 times (p<0.01), 12.2 times (p<0.01) and 8.6 times (p<0.01), respectively. Also, both the prophylactic and therapeutic use of 4-hydroxy-3,5-di-tret-butyl cinnamic acid normalized aerobic/anaerobic metabolism, as well as reduced the concentration of caspase-3. Based on the obtained data, significant cerebroprotective properties of 4-hydroxy-3,5- di-tertbutyl cinnamic acid can be assumed. Moreover, the potential mechanism of action of this compound may be mediated by the normalization of mitochondrial function.

Hypoxia Induced Infertility in Males: Role of Transcription Factor Hypoxia Inducible Factor -1 Alpha

Aim: To assess concentrations of HIF-1 α and correlation between HIF-1 α in healthy and sub fertile males’ population in our study. Methodology: In this study we investigated the concentrations of Hypoxia Inducible Factor 1-α (pg/mL) in the seminal plasma of healthy fertile males and sub fertile males using Enzyme-Linked ImmunoSorbent Assay (ELISA). Our study population (n=54) consisted of healthy fertile controls (n=18), sub fertile males without varicocele (n=22) and sub fertile males with varicocele. Result: Our results showed significantly (p <0.001) elevated levels of HIF 1-α in both sub fertile groups (p< 0.001) as compared to healthy fertile group. A significant (r= 0.975, p<0.001) positive correlation was noticed between the concentrations of Caspase 3, an apoptotic marker and HIF 1-α in the healthy fertile and sub fertile groups. Conclusions: Our study results suggest that hypoxia induced apoptosis maybe an important factor in causing testicular dysfunction in sub fertile males. In conclusion HIF 1-α is an important hypoxic factor that can be used to predict apoptosis in testes. HIF 1-α can be used as a clinical marker that can facilitate scientists to predict the degree of apoptosis in spermatozoa. Keywords: HIF-1, Subfertile, Apoptotic marker, Spermatozoa

Increased expression of caspase 1 during active phase of connective tissue disease.

Key factors of pyroptosis play an important role in the inflammatory response to connective tissue disease (CTD). However, information on active and stable stages of CTD is scarce. To distinguish the differences of concentrations of C-reactive protein (CRP), caspase 1, caspase 4, caspase 5 and sCD14 in plasma between the patients with active and stable stages of CTD. A cohort study was conducted to recruit patients diagnosed with CTD of active phase and stable phase as well as health control. These data included the analysis of the concentration of sCD14, caspase 1, caspase 4 and caspase 5 in peripheral plasma by ELISA. The Wilcoxon rank-sum test was used to compare the two groups. The sex ratio and ages of the three groups were not different statistically. The concentrations of sCD14, caspase4 and caspase5 of plasma in the CTD of active phase and the stable phase as well as the health control. The concentration of caspase 1 in active phase of CTD (470.19 [422.33–513.14] pmol/L) was significantly higher than that in stable group (203.95 [160.94–236.12] pmol/L) and healthy control (201.65 [191.11–240.35] pmol/L] pmol/L) (p < 0.001, both), but there was no significant difference between stable group and healthy control (p = 0.2312). Similarly, the concentration of CRP in the active phase of CTD (8.96 [3.06–20.28] mg/L) was significantly higher than that in the stable group (3.00 [1.30–11.40] mg/L) and the healthy control (3.70 [2.30–4.73] mg/L) (p = 0.0013, p = 0.0006, respectively), but there was no significant difference between the stable group and the healthy control (p = 0.3205). However, there were no significant differences in the concentration of sCD14, caspase 4 and caspase 5 in the active phase of CTD and the stable group as well as the health group. Consequently, the patients of the active phase of CTD showed increased expression of caspase 1.

Role of Mitochondria in Ciprofloxacin-Induced Apoptosis in Murine Sperm Cells

Ciprofloxacin (CPFX) has been reported to inhibit cell growth and induce apoptosis in certain eukaryotic cells. The role of the mitochondrial pathway in CPFX-induced apoptosis in cultured murine sperm cells was investigated. Sperm cells (5×10(3) cells/well) from 8-week-old NMRI male mice were cultured in 150 μL of HAM's F10 with 25 mmol/L (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid ) HEPES and 10% human serum albumin and were incubated with 50, 100, 200, 400, and 800 µg/mL CPFX for 24 and 36 hours. Cell cytotoxicity, mitochondrial membrane potential (ΔΨM), and concentrations of caspase 3 and caspase 9 were assessed in CPFX-treated cultured murine sperm cells by MTT (3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide) , JC-1 (5, 5á, 6, 6á-tetrachloro-1, 1á, 3, 3á-tetraethylbenzimidazol-carbocyanine iodide)aggregation, and caspase 3 and caspase 9 assays, respectively. Increasing doses of CPFX showed significant cytotoxicity (EC50 = 146.73 μg/mL). Significant loss of ΔΨm was observed in sperm cells treated with ≥50 μg/mL CPFX for 36 hours. Significant increases in caspase 9 and caspase 3 concentrations were also found in cells treated with ≥50 μg/mL CPFX for 24 and 36 hours, respectively (P < .001). Effects of clinically reachable doses of CPFX on cultured murine sperm cells were investigated and revealed that it may cause sperm cell toxicity by induction of apoptosis through the mitochondrial pathway in the clinically reachable concentrations.

Effect of N-acetyl cysteine on enterocyte apoptosis and intracellular signalling pathways' response to oxidative stress in weaned piglets

N-Acetyl cysteine (NAC) has been widely used for preventing reactive oxygen species-induced damage. However, little is known as to whether dietary NAC supplementation would alleviate intestinal injury in weaned piglets. The present study evaluated the effect of NAC on enterocyte apoptosis and intracellular signalling pathways' response to weaning stress. The control piglets were normally suckling, and piglets in the weaning and NAC groups were fed the basal diet and basal+NAC diet from 14 to 25 d of age, respectively. Compared with the control piglets, weaning increased cortisol concentrations (P< 0·05), decreased superoxide dismutase and glutathione peroxidase activities (P< 0·05), increased malondialdehyde content (P< 0·05) in serum and enhanced enterocyte apoptosis index (AI) and concentrations of caspase-3, caspase-8 and caspase-9 (P< 0·05). Gene expression analyses indicated that weaning induced apoptosis via Fas signalling and mitochondrial pathways in weaned piglets. Dietary NAC supplementation decreased (P< 0·05) cortisol concentrations and the AI, increased (P< 0·05) antioxidant status in serum and alleviated histopathological changes in the intestine. It also inhibited Fas, caspase-3, caspase-8 and integrin αvβ6 (αvβ6) gene expressions in the NAC-treated piglets. However, no significant decrease (P>0·10) in caspase-3, caspase-8 and caspase-9 concentrations was observed in the NAC group compared with the weaning group. In conclusion, weaning may induce enterocyte apoptosis via the activation of Fas-dependent and mitochondria-dependent apoptosis. Although NAC had no effect on caspase concentrations, it was clearly beneficial for preserving morphological integrity in weaned piglets via the regulation of cell apoptosis and the inhibition of Fas-dependent apoptosis and αvβ6 expression.

XIAP As a Positive Feedback Regulator of Capase Activation

STKE Inhibitor of apoptosis proteins (IAPs) are best known for their roles as inhibitors of caspases and thus of apoptotic cell death. Legewie et al. develop a mathematical model of the core intrinsic apoptotic process involving Apaf-1, caspase 9, caspase 3, and X-linked IAP (XIAP). In the wild-type model, as active Apaf-1 concentration increased, the time course with which active caspase 3 was produced decreased as expected. When the activation of caspase 3 was plotted against the concentration of active Apaf-1, a bistable and irreversible state was observed both in the wild-type model and in a caspase 9 mutant model. This suggested that the positive feedback between caspase 9 and caspase 3 was not the only contributor to bistability in the system. Further analysis suggested that XIAP contributed a positive feedback to caspase 3 activation. Caspase 3 was proposed to sequester XIAP away from caspase 9 under conditions of strong stimulation, thereby allowing caspase 9 to become further activated and ultimately allowing caspase 3 to be activated. The bistability and irreversibility of the models depended on the concentrations of both caspase 3 and caspase 9, with only the wild-type model showing irreversibility in the physiological concentrations of each caspase, indicating that both caspase 3-mediated feedback and XIAP-mediated feedback contribute to irreversible bistability under physiological conditions. The models echo observed differences in cellular responses to apoptotic stimuli by showing that the all-or-none threshold of the system is influenced by the abundance of caspase 3, caspase 9, and XIAP. — NRG PLoS Comput. Biol. 2 , e120 (2006).

XIAP as a Positive Feedback Regulator of Caspase Activation

Inhibitor of apoptosis proteins (IAPs) are best known for their roles as inhibitors of caspases and thereby inhibitors of the cell death process known as apoptosis. Legewie et al . developed a mathematical model of the core intrinsic apoptotic process involving Apaf-1, caspase 9, caspase 3, and X-linked IAP (XIAP). The input into the model is increased abundance of cytochrome c, which activates Apaf-1e, and the output is the concentration of active caspase 3. The model is based on 16 kinetic parameters and 13 differential equations. Three models were tested: a wild-type model in which XIAP competitively interacts with caspase 3 and caspase 9, a model in the presence of a mutant caspase 9 that cannot be cleaved and activated by caspase 3, and a noncompetitive model in which XIAP noncompetitively interacted with both caspase 3 and caspase 9. In the wild-type model, as active Apaf-1 concentration increased, the time course with which active caspase 3 was produced decreased as expected. When the activation of caspase 3 was plotted against the concentration of active Apaf-1, a bistable and irreversible state was observed both in the wild-type model and in the caspase 9 mutant model (although the total concentration of active caspase 3 was less in the caspase 9 mutant model). This suggested that the positive feedback between caspase 9 and caspase 3 was not the only contributor to bistability in the system. Further analysis suggested that XIAP contributed a positive feedback into caspase 3 activation. Caspase 3 was proposed to sequester XIAP away from caspase 9 under conditions of strong stimulation, thereby allowing caspase 9 to become further activated and ultimately allowing caspase 3 to be activated. The requirements were that the interactions between XIAP and caspase 3 and caspase 9 were competitive, caspase 3 abundance exceeded that of XIAP, and XIAP abundance exceeded that of caspase 9. The bistability and irreversibility of all three models depended on the concentrations of both caspase 3 and caspase 9, with only the wild-type model showing irreversibility in the physiological concentrations of each caspase, indicating that both caspase 3-mediated feedback and XIAP-mediated feedback contribute to irreversible bistability under physiological conditions. Their model accounts for experimentally observed differences in cellular responses to apoptotic stimuli by showing that the all-or-none threshold of the system is influenced by the abundance of caspase 3, caspase 9, and XIAP. The authors suggest wet-lab experiments that could be used to verify the model proposed. S. Legewie, N. Blüthgen, H. Herzel, Mathematical modeling identifies inhibitors of apoptosis as mediators of positive feedback and bistability. PLoS Comput. Biol. 2 , e120, 1061-1073 (2006). [PubMed]

Apoptotic neuronal death following deep hypothermic circulatory arrest in piglets.

Deep hypothermic circulatory arrest (DHCA), as used in infant heart surgery, carries a risk of brain injury. In a piglet DHCA model, neocortical neurons appear to undergo apoptotic death. Caspases, cytochrome c, tumor necrosis factor (TNF), and Fas play a role in apoptosis in many ischemic models. This study examined the expression of these factors in a DHCA piglet model. Thirty-nine anesthetized piglets were studied. After cardiopulmonary bypass (CPB) cooling of the brain temperature to 19 degrees C, DHCA was induced for 90 min, followed by CPB rewarming. After separation from CPB, piglets were killed at 1, 4, 8, 24, and 72 h and 1 week. Caspase-8 and -3 activity, and concentrations of TNF-alpha, Fas, Fas-ligand, cytochrome c, and adenosine triphosphate (ATP) were measured in the neocortex by enzymatic assay and Western blot analysis. Caspase-8 and -3 activity and cell death were examined histologically. Significance was set at P < 0.05. In neocortex, damaged neurons were not observed in control (no CPB), rarely observed in CPB (no DHCA), and rarely observed in the DHCA 1-h, 4-h, and 1-week reperfusion groups. However, they were seen frequently in the DHCA 8-, 24-, and 72-h reperfusion groups. Although neuronal death was widespread 8-72 h after DHCA, cortical ATP concentrations remained unchanged from control. Both caspase-3 and -8 activities were significantly increased at 8 h after DHCA, and caspase-3 concentration remained elevated for as long as 72 h. Caspase-3 and -8 activity was also observed in damaged neocortical neurons. Cytosolic cytochrome c and Fas were significantly expressed at 1 h and 4 h after DHCA, respectively. Fas-ligand and TNF-alpha were not observed in any group. After DHCA, induction of apoptosis in the neocortex occurs within a few hours of reperfusion and continues for several days. Increased Fas, cytochrome c, and caspase concentrations, coupled with normal brain ATP concentrations and apoptotic histologic appearance, are consistent with the occurrence of apoptotic cell death.