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#33654958   2019/12/20 To Up

Isolation and Stimulation of Peritoneal Macrophages withApoptotic Jurkat Cells to Produce IL-10.

Clearance of apoptotic cells by macrophages is critical to ensuring cellular homeostasis and suppression of autoimmunity. Macrophage recognition of apoptotic cells triggers an anti-inflammatory response, which is mediated by the release of IL-10, TGF-β . with concurrent inhibition of pro-inflammatory cytokines (such as TNFα, IL-12, IL-1β). To characterize cytokine profile produced by macrophages during phagocytosis of apoptotic cells, we developed an effective, more physiologic system using isolated murine peritoneal macrophages and T-lymphocyte cell line Jurkat as a source of apoptotic cells. Apoptosis of Jurkat cells is induced with staurosporine, a protein kinase C (PKC) inhibitor and detected by Annexin V/propidium iodide staining. This in assay demonstrates that murine peritoneal macrophages produce large amounts of IL-10 following exposure to apoptotic Jurkat cells.
Mei Song, Xiaojing Ma

2267 related Products with: Isolation and Stimulation of Peritoneal Macrophages withApoptotic Jurkat Cells to Produce IL-10.

100 µg100 extractions100 µg100ul1 mg100ul10 1.00 flask1mg4 X 250 ml.2 x 10^6 cells

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#33649872   2021/03/01 To Up

Feeding behavior of feedlot-finished crossbred bulls fed palm kernel cake.

The aim of this study was to evaluate the effects of dietary inclusion of palm kernel cake (PKC) on the feeding behavior of feedlot-finished cattle. Forty uncastrated crossbred bulls with an average age of 24.2 ± 2 months and an average weight of 331.1 ± 36.2 kg were distributed into four treatment groups in a completely randomized design with ten replicates. The treatments consisted of 0, 8, 16, or 24% PKC inclusion in the diet (dry matter basis). Palm kernel cake inclusion influenced feeding time (P < 0.05), which increased linearly. The inclusion of the ingredient also affected (P < 0.05) rumination time (min/day), total chewing time, and the time spent on other activities. The number of cuds ruminated per day showed a linear response to the PKC inclusion levels (P < 0.05). The number of feeding bouts increased linearly (P < 0.05). Dry matter (DM) intake and DM feeding efficiency decreased linearly (P < 0.05). There were no effects of PKC inclusion (P > 0.05) on neutral detergent fiber feeding (NDF) efficiency or DM rumination efficiency. NDF rumination efficiency increased linearly (P < 0.05). Palm kernel cake can be included at levels up to 24% in the total diet of feedlot-finished crossbred bulls, as it does not affect their dry matter intake or feeding behavior.
Mateus de M Lisboa, Robério R Silva, Fabiano F da Silva, Maria M S Pereira, Gabriel D Costa, Fabrício B L Mendes, Sinvaldo O de Souza, Marceliana da C Santos, Laoan B O Rodrigues, Henry D R Alba, Gleidson G P de Carvalho

1435 related Products with: Feeding behavior of feedlot-finished crossbred bulls fed palm kernel cake.

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#33645025   // To Up

[Cloning and expression analysis of protein kinase C gene from Whitmania pigra].

Protein kinase C(PKC) is a type of protein kinase widely involved in cell proliferation and development, but the developmental mechanism in the gonads of androgynous animals is still unclear. In order to explore the role of protein kinase C in the development of Whitmania pigra germ cells, the Wh. pigra PKC(Wp-PKC) gene was cloned, bioinformatics analysis was conducted, and fluorescent quantitative PCR was used to analyze the expression of female and male gonads. The results showed that:(1)The cloned Wp-PKC had a full length of 2 580 bp, a relative molecular weight of 76 555.19, and contains an open reading frame encoding 670 amino acids, Wp-PKC was closely related to Danio rerio PKC-α and rat PKC-γ. The similarity of amino acid sequence was 55% and 58%.(2)The protein encoded by Wp-PKC had no signal peptide and was a hydrophilic protein. The secondary structure is mainly composed of random coils, α-helices, extended chains, folds and folds, with the largest proportion of random coils and α-helices. Wp-PKC protein does not contain a transmembrane domain. Multiple sequence alignment and domain prediction analysis show that Wp-PKC contains 4 conserved domains of classical protein kinase C.(3)Fluorescence quantitative results showed that the expression of Wp-PKC in Wh. pigra gonads was positively correlated with the development of germ cells, and the expression in male gonads was significantly higher than that in female gonads. In summary, Wp-PKC is a classic PKC, and Wp-PKC may promote the development of Wh. pigra, especially the development of male gonads, and provide references for further research on the developmental mechanisms of Wh. pigra.
Pan-Pan Chen, Qiao-Sheng Guo, Hong-Zhuan Shi, Li-Yuan Guo, Hui-Ning Zhang

1940 related Products with: [Cloning and expression analysis of protein kinase C gene from Whitmania pigra].

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#33643814   2020/09/19 To Up

Phosphorylation regulates cullin-based ubiquitination in tumorigenesis.

Cullin-RING ligases (CRLs) recognize and interact with substrates for ubiquitination and degradation, and can be targeted for disease treatment when the abnormal expression of substrates involves pathologic processes. Phosphorylation, either of substrates or receptors of CRLs, can alter their interaction. Phosphorylation-dependent ubiquitination and proteasome degradation influence various cellular processes and can contribute to the occurrence of various diseases, most often tumorigenesis. These processes have the potential to be used for tumor intervention through the regulation of the activities of related kinases, along with the regulation of the stability of specific oncoproteins and tumor suppressors. This review describes the mechanisms and biological functions of crosstalk between phosphorylation and ubiquitination, and most importantly its influence on tumorigenesis, to provide new directions and strategies for tumor therapy.
Yifan Chen, Xuejing Shao, Ji Cao, Hong Zhu, Bo Yang, Qiaojun He, Meidan Ying

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#33641369   2021/03/01 To Up

EP3 (E-Prostanoid 3) Receptor Mediates Impaired Vasodilation in a Mouse Model of Salt-Sensitive Hypertension.

We previously showed that impaired vasodilation in systemic and renal vessels contributes to salt-sensitive hypertension in a mouse model of impaired PPARγ (peroxisome proliferator-activated receptor gamma) function. We determined the mechanisms mediating impaired salt-induced vasodilation and whether improved vasodilation attenuates augmented hypertension in response to salt. Mice selectively expressing a PPARγ dominant negative mutation in vascular smooth muscle (S-P467L) exhibited salt-sensitive hypertension and severely impaired vasodilation in systemic and renal vessels. High-salt diet-fed S-P467L and control mice displayed comparable levels of renal oxidative stress markers. Preincubation with Tempol, a superoxide dismutase mimetic, or calphostin C, a PKC (protein kinase C) inhibitor, failed to improve salt-induced impairment of vasodilation in S-P467L mice, arguing against a role of oxidative stress or PKC activity. Inhibition of Rho kinase partially rescued impaired vasodilation in high-salt diet-fed S-P467L mice suggesting a contribution of the Ras homolog family member A (RhoA)/Rho kinase pathway. High-salt diet selectively increased synthesis of PGE2 (prostaglandin E2) in S-P467L aorta. Expression of EP3 (E-prostanoid 3) receptor mRNA was increased in aorta from chow-fed and high salt-fed S-P467L mice. Pharmacological inhibition of COX (cyclooxygenase) 2 or blockade of EP3 completely normalized the impaired vasodilation, and EP3 antagonism induced larger decreases in systolic blood pressure in high-salt diet-fed S-P467L mice. In conclusion, interference with PPARγ in vascular smooth muscle causes activation of the PGE2/EP3 signaling pathway in systemic and renal vasculature resulting in salt-induced impairment of vasodilation and salt-sensitive hypertension. PGE2/EP3 axis maybe a druggable target to prevent salt-sensitive hypertension in chronic conditions associated with decreased PPARγ activity.
Jing Wu, Shi Fang, Ko-Ting Lu, Kelsey Wackman, Michal L Schwartzman, Sergey I Dikalov, Justin L Grobe, Curt D Sigmund

1848 related Products with: EP3 (E-Prostanoid 3) Receptor Mediates Impaired Vasodilation in a Mouse Model of Salt-Sensitive Hypertension.

100 μg100 μg500 100.00 ug100.00 ug100 μg100 ul4 Arrays/Slide50 1mg1 mg100 μg

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#33641357   2021/03/01 To Up

TRPV1 (Transient Receptor Potential Vanilloid 1) Sensitization of Skeletal Muscle Afferents in Type 2 Diabetic Rats With Hyperglycemia.

The blood pressure response to exercise is exaggerated in type 2 diabetes (T2D). However, the underlying mechanisms remain unclear. It is hypothesized that one mechanism mediating the potentiated cardiovascular response in T2D is the sensitization of chemically sensitive afferent neurons by activation of metaboreceptors. To test this hypothesis, we examined TRPV1 (transient receptor potential vanilloid 1)-induced cardiovascular responses in vivo and muscle afferent discharge ex vivo in T2D rats. Additionally, TRPV1 and PKC (protein kinase C) protein levels in dorsal root ganglia subserving skeletal muscle were assessed. For 14 to 16 weeks, Sprague-Dawley rats were given either a normal diet (control) or a high-fat diet in combination with a low dose (35 and 25 mg/kg) of streptozotocin (T2D). Administration of capsaicin, TRPV1 agonist, in hindlimb evoked significantly greater increases in mean arterial pressure and renal sympathetic nerve activity in decerebrated T2D than control. In a muscle-nerve preparation, the discharge to capsaicin exposure in group IV afferents isolated from T2D was likewise significantly augmented at a magnitude that was proportional to glucose concentration. Moreover, the discharge to capsaicin was potentiated by acute exposure of group IV afferents to a high-glucose environment. T2D showed significantly increased phospholyrated-TRPV1 and -PKCα levels in dorsal root ganglia neurons as compared with control. These findings suggest that group IV muscle afferents are sensitized by PKC-induced TRPV1 overactivity in early stage T2D with hyperglycemia and, thereby, may contribute to the potentiated circulatory response to TRPV1 activation in the disease.
Rie Ishizawa, Han-Kyul Kim, Norio Hotta, Gary A Iwamoto, Jere H Mitchell, Scott A Smith, Wanpen Vongpatanasin, Masaki Mizuno

1767 related Products with: TRPV1 (Transient Receptor Potential Vanilloid 1) Sensitization of Skeletal Muscle Afferents in Type 2 Diabetic Rats With Hyperglycemia.

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#33637129   2021/02/26 To Up

Dapagliflozin attenuates hypoxia/reoxygenation-caused cardiac dysfunction and oxidative damage through modulation of AMPK.

Emerging evidence demonstrated dapagliflozin (DAPA), a sodium-glucose cotransporter 2 inhibitor, prevented various cardiovascular events. However, the detailed mechanisms underlying its cardioprotective properties remained largely unknown.
Kun-Ling Tsai, Pei-Ling Hsieh, Wan-Ching Chou, Hui-Ching Cheng, Yu-Ting Huang, Shih-Hung Chan

2462 related Products with: Dapagliflozin attenuates hypoxia/reoxygenation-caused cardiac dysfunction and oxidative damage through modulation of AMPK.

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#33626581   2021/02/18 To Up

Characterization of the Platelet Phenotype Caused by a Germline RUNX1 Variant in a CRISPR/Cas9-Generated Murine Model.

-related disorder (-RD) is caused by germline variants affecting the gene. This rare, heterogeneous disorder has no specific clinical or laboratory phenotype, making genetic diagnosis necessary. Although international recommendations have been established to classify the pathogenicity of variants, identifying the causative alteration remains a challenge in -RD. Murine models may be useful not only for definitively settling the controversy about the pathogenicity of certain variants, but also for elucidating the mechanisms of molecular pathogenesis. Therefore, we developed a murine model, using the CRISPR/Cas9 system, carrying the RUNX1 p.Leu43Ser variant (mimicking human p.Leu56Ser) to study its pathogenic potential and mechanisms of platelet dysfunction. A total number of 75 mice were generated; 25 per genotype (RUNX1, RUNX1, and RUNX1). Platelet phenotype was assessed by flow cytometry and confocal microscopy. On average, RUNX1 and RUNX1 mice had a significantly longer tail-bleeding time than RUNX1 mice, indicating the variant's involvement in hemostasis. However, only homozygous mice displayed mild thrombocytopenia. RUNX1 and RUNX1 displayed impaired agonist-induced spreading and α-granule release, with no differences in δ-granule secretion. Levels of integrin αβ activation, fibrinogen binding, and aggregation were significantly lower in platelets from RUNX1 and RUNX1 using phorbol 12-myristate 13-acetate (PMA), adenosine diphosphate (ADP), and high thrombin doses. Lower levels of PKC phosphorylation in RUNX1 and RUNX1 suggested that the PKC-signaling pathway was impaired. Overall, we demonstrated the deleterious effect of the RUNX1 p.Leu56Ser variant in mice via the impairment of integrin αβ activation, aggregation, α-granule secretion, and platelet spreading, mimicking the phenotype associated with variants in the clinical setting.
Ana Marín-Quílez, Ignacio García-Tuñón, Cristina Fernández-Infante, Luis Hernández-Cano, Verónica Palma-Barqueros, Elena Vuelta, Manuel Sánchez-Martín, José Ramón González-Porras, Carmen Guerrero, Rocío Benito, José Rivera, Jesús María Hernández-Rivas, José María Bastida

2643 related Products with: Characterization of the Platelet Phenotype Caused by a Germline RUNX1 Variant in a CRISPR/Cas9-Generated Murine Model.

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#33626379   2021/02/21 To Up

Protein kinase C-mediated calcium signaling as the basis for cardiomyocyte plasticity.

Protein kinase C is the superfamily of intracellular effector molecules which control crucial cellular functions. Here, we for the first time did the percentage estimation of all known PKC and PKC-related isozymes at the individual cadiomyocyte level. Broad spectrum of PKC transcripts is expressed in the left ventricular myocytes. In addition to the well-known 'heart-specific' PKCα, cardiomyocytes have the high expression levels of PKCN1, PKCδ, PKCD2, PKCε. In general, we detected all PKC isoforms excluding PKCη. In cardiomyocytes PKC activity tonically regulates voltage-gated Ca-currents, intracellular Ca level and nitric oxide (NO) production. Imidazoline receptor of the first type (IR)-mediated induction of the PKC activity positively modulates Ca release through ryanodine receptor (RyR), increasing the Ca leakage in the cytosol. In cardiomyocytes with the Ca-overloaded regions of > 9-10 μm size, the local PKC-induced Ca signaling is transformed to global accompanied by spontaneous Ca waves propagation across the entire cell perimeter. Such switching of Ca signaling in cardiac cells can be important for the development of several cardiovascular pathologies and/or myocardial plasticity at the cardiomyocyte level.
Alexander V Maltsev, Edward V Evdokimovskii, Yury M Kokoz

1564 related Products with: Protein kinase C-mediated calcium signaling as the basis for cardiomyocyte plasticity.

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