Diabetic kidney disease (DKD) is the leading cause of end-stage renal disease. Therefore, understanding the molecular regulatory mechanisms underlying the pathogenesis of DKD is imperative. In this study, we aimed to explore the molecular mechanisms of tubule region endothelial dysfunction in early DKD. Early-stage DKD model was established in 16-week-old female db/db mice for 16 weeks. Body weight, glucose level, and urine albumin-to-creatinine ratio (UACR) were measured. Hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) staining were performed to evaluate pathological lesions. RNA sequencing data of the kidneys and integrated publicly available single-cell and spatial transcriptome datasets were used to investigate the mechanism of endothelial dysfunction. There was a significant increase in body weight (p = 0.001), glucose levels (p=0.0008), and UACR (p=0.006) in db/db mice compared with db/m mice. H&E and PAS staining showed that vacuolar lesions and protein casts of tubules were the major histopathological changes observed in early-stage DKD mice. The apoptotic pathway in endothelial cells was notably activated in DKD, and Thbs1 was identified as the central gene involved in this apoptotic process. Deconvolution of the cell composition in the RNA sequencing data showed a decrease in the proportion of endothelial cells in the DKD mice. Further analysis of the activity and regulatory network of transcription factors showed that Creb1 was activated in both mouse and human early-stage DKD, suggesting that Creb1 activation may be involved in early kidney injury. The endothelial cell apoptotic pathway is activated in DKD, and the proportion of endothelial cells was reduced in the DKD mice, which is significantly associated with Thbs1. Keywords: Diabetic kidney disease, Endothelial dysfunction, RNA sequencing,Thbs1, Creb1.

• MeSH
• apoptóza MeSH
• diabetické nefropatie * patologie   metabolismus   patofyziologie   genetika   MeSH
• endoteliální buňky metabolismus   patologie   MeSH
• ledvinové kanálky patologie   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• progrese nemoci * MeSH
• protein vázající cAMP responzivní element metabolismus   genetika   MeSH
• thrombospondin 1 metabolismus   genetika   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The mucus layer protects airway epithelia from damage by noxious agents. Intriguingly, Bordetella pertussis bacteria provoke massive mucus production by nasopharyngeal epithelia during the initial coryza-like catarrhal stage of human pertussis and the pathogen transmits in mucus-containing aerosol droplets expelled by sneezing and post-nasal drip-triggered cough. We investigated the role of the cAMP-elevating adenylate cyclase (CyaA) and pertussis (PT) toxins in the upregulation of mucin production in B. pertussis-infected airway epithelia. Using human pseudostratified airway epithelial cell layers cultured at air-liquid interface (ALI), we show that purified CyaA and PT toxins (100 ng/mL) can trigger production of the major airway mucins Muc5AC and Muc5B. Upregulation of mucin secretion involved activation of the cAMP response element binding protein (CREB) and was blocked by the 666-15-Calbiochem inhibitor of CREB-mediated gene transcription. Intriguingly, a B. pertussis mutant strain secreting only active PT and producing the enzymatically inactive CyaA-AC- toxoid failed to trigger any important mucus production in infected epithelial cell layers in vitro or in vivo in the tracheal epithelia of intranasally infected mice. In contrast, the PT- toxoid-producing B. pertussis mutant secreting the active CyaA toxin elicited a comparable mucin production as infection of epithelial cell layers or tracheal epithelia of infected mice by the wild-type B. pertussis secreting both PT and CyaA toxins. Hence, the cAMP-elevating activity of B. pertussis-secreted CyaA was alone sufficient for activation of mucin production through a CREB-dependent mechanism in B. pertussis-infected airway epithelia in vivo.

• MeSH
• adenylátcyklasový toxin toxicita   MeSH
• Bordetella pertussis metabolismus   patogenita   MeSH
• buněčné linie MeSH
• dýchací soustava metabolismus   mikrobiologie   MeSH
• epitelové buňky metabolismus   mikrobiologie   MeSH
• lidé MeSH
• mucin 5AC metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• pertuse metabolismus   mikrobiologie   MeSH
• protein vázající cAMP responzivní element metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Controlling pain in burn-injured patients poses a major clinical challenge. Recent findings suggest that reducing the activity of the voltage-gated sodium channel Nav1.7 in primary sensory neurons could provide improved pain control in burn-injured patients. Here, we report that partial thickness scalding-type burn injury on the rat paw upregulates Nav1.7 expression in primary sensory neurons 3 h following injury. The injury also induces upregulation in phosphorylated cyclic adenosine monophosphate response element-binding protein (p-CREB), a marker for nociceptive activation in primary sensory neurons. The upregulation in p-CREB occurs mainly in Nav1.7-immunopositive neurons and exhibits a peak at 5 min and, following a decline at 30 min, a gradual increase from 1 h post-injury. The Nav1.7 blocker protoxin II (ProTxII) or morphine injected intraperitoneally 15 min before or after the injury significantly reduces burn injury-induced spinal upregulation in phosphorylated serine 10 in histone H3 and phosphorylated extracellular signal-regulated kinase 1/2, which are both markers for spinal nociceptive processing. Further, ProTxII significantly reduces the frequency of spontaneous excitatory post-synaptic currents in spinal dorsal horn neurons following burn injury. Together, these findings indicate that using Nav1.7 blockers should be considered to control pain in burn injury. KEY MESSAGES: • Burn injury upregulates Nav1.7 expression in primary sensory neurons. • Burn injury results in increased activity of Nav1.7-expressing primary sensory neurons. • Inhibiting Nav1.7 by protoxin II reduces spinal nociceptive processing. • Nav1.7 represents a potential target to reduce pain in burn injury.

• MeSH
• analgetika terapeutické užití   MeSH
• blokátory sodíkových kanálů řízených napětím terapeutické užití   MeSH
• bolest farmakoterapie   MeSH
• mícha cytologie   fyziologie   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• napěťově řízený sodíkový kanál, typ 9 fyziologie   MeSH
• nervové receptory fyziologie   MeSH
• pavoučí jedy terapeutické užití   MeSH
• peptidy terapeutické užití   MeSH
• popálení farmakoterapie   MeSH
• potkani Sprague-Dawley MeSH
• potkani Wistar MeSH
• protein vázající cAMP responzivní element metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Prenatal exposure to caffeine can cause developmental problems. This study determined chronic influence of prenatal caffeine at relatively higher doses on cognitive functions in the rat offspring. Pregnant Sprague-Dawley rats (4-month-old) were exposed to caffeine (20 mg/kg, twice a day) for whole pregnancy from gestational day 4. Fetal and offspring body and brain weight was measured. Learning and memory were tested in adult offspring with Morris water maze. Learning and memory-related receptors were measured. The exposure to prenatal caffeine not only caused fetal growth restriction, but also showed long-term effects on learning and memory in the offspring. The caffeine offspring exhibited longer escape latency and path length in navigation testing. The number of passing the target was significantly reduced in those offspring. The expression of adenosine A(1) and A(2A) receptors, nuclear PKA C(alpha), C(beta) subunits, and pCREB were significantly increased in the fetal and neonatal brain, and suppressed in the hippocampus of the adult offspring. The expression of BDNF and TrkB were reduced regardless of various ages. The results suggest that intrauterine programming dysfunction of adenosine receptors and the down-stream of cAMP/PKA/pCREB system may play an important role in prenatal caffeine induced cognition disorders in the adult offspring.

• MeSH
• bludiště - učení účinky léků   fyziologie   MeSH
• kofein toxicita   MeSH
• krysa rodu rattus MeSH
• mozkový neurotrofický faktor metabolismus   MeSH
• paměť účinky léků   fyziologie   MeSH
• poruchy paměti chemicky indukované   diagnóza   metabolismus   MeSH
• potkani Sprague-Dawley MeSH
• protein vázající cAMP responzivní element metabolismus   MeSH
• proteinkinasy závislé na cyklickém AMP metabolismus   MeSH
• purinergní receptory P1 metabolismus   MeSH
• signální transdukce účinky léků   fyziologie   MeSH
• těhotenství MeSH
• zpožděný efekt prenatální expozice chemicky indukované   diagnóza   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Chronic exposure to low dose of T-2 toxin causes growth retardation and reduced body weight, resulting in economic losses. Excessive inflammatory cytokines and GH deficiency are important mechanisms that contribute to growth inhibition induced by T-2 toxin. However, the regulation of the inflammatory cytokines expecially IL-6, IL-1β, and TNF-α induced by T-2 toxin still remained unclear. The new transcription factor AKNA, belonging to AT-hook protein family, is closely associated with inflammation. However, it was unclear how AKNA regulate the expression of inflammatory cytokines, and there was no report on the role of AKNA in T-2 toxin mediated toxicity. Here, we investigated the role of AKNA in T-2 toxin-mediated inflammatory response and GH deficiency and the signal transduction pathway of AKNA. We showed that AKNA regulated by PKA/CREB and NF-κB pathway is a novel downstream molecular target in T-2 toxin-mediated inflammation and GH deficiency. T-2 toxin activates the PKA/CREB and NF-κB/p65 pathways, thereby promoting the direct binding of phospho-CREB and phospho-p65 to the AKNA promoter, thus inhibiting AKNA expression. GH and inflammatory cytokines (TNF-α, IL-1β, and IL-6) expression were significantly downregulated after AKNA silencing. Furthermore, the expression of differential genes induced by T-2 toxin in the rat pituitary was further confirmed by acute toxicity tests in rats, which was consistent with the results in GH3 cells. By histopathological analysis, we confirmed the pituitary might be a novel direct target organ of T-2 toxin. These findings provided new insights into the significant role of AKNA in T-2 toxin-induced inflammatory response and growth inhibition.

• MeSH
• buněčné linie MeSH
• DNA vazebné proteiny genetika   metabolismus   MeSH
• down regulace MeSH
• fosforylace MeSH
• interleukin-1beta genetika   metabolismus   MeSH
• interleukin-6 genetika   metabolismus   MeSH
• jaderné proteiny genetika   metabolismus   MeSH
• krysa rodu rattus MeSH
• NF-kappa B metabolismus   MeSH
• promotorové oblasti (genetika) MeSH
• protein vázající cAMP responzivní element metabolismus   MeSH
• proteinkinasy závislé na cyklickém AMP metabolismus   MeSH
• T-2 toxin toxicita   MeSH
• TNF-alfa genetika   metabolismus   MeSH
• transkripční faktory genetika   metabolismus   MeSH
• umlčování genů MeSH
• zánět chemicky indukované   genetika   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Monitoring of single cell signal transduction in leukemic cellular subsets has been proposed to provide deeper understanding of disease biology and prognosis, but has so far not been tested in a clinical trial of targeted therapy. We developed a complete mass cytometry analysis pipeline for characterization of intracellular signal transduction patterns in the major leukocyte subsets of chronic phase chronic myeloid leukemia. Changes in phosphorylated Bcr-Abl1 and the signaling pathways involved were readily identifiable in peripheral blood single cells already within three hours of the patient receiving oral nilotinib. The signal transduction profiles of healthy donors were clearly distinct from those of the patients at diagnosis. Furthermore, using principal component analysis, we could show that phosphorylated transcription factors STAT3 (Y705) and CREB (S133) within seven days reflected BCR-ABL1IS at three and six months. Analyses of peripheral blood cells longitudinally collected from patients in the ENEST1st clinical trial showed that single cell mass cytometry appears to be highly suitable for future investigations addressing tyrosine kinase inhibitor dosing and effect. (clinicaltrials.gov identifier: 01061177).

• MeSH
• analýza jednotlivých buněk metody   MeSH
• bcr-abl fúzní proteiny metabolismus   MeSH
• chronická fáze myeloidní leukemie farmakoterapie   patologie   MeSH
• fosforylace MeSH
• leukocyty metabolismus   MeSH
• lidé MeSH
• protein vázající cAMP responzivní element metabolismus   MeSH
• pyrimidiny farmakologie   terapeutické užití   MeSH
• signální transdukce účinky léků   imunologie   MeSH
• transkripční faktor STAT3 metabolismus   MeSH
• tyrosinkinasy terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky MeSH
• práce podpořená grantem MeSH

The aim of our study was to examine the in vivo expression of p21 and expression and activation of ATF-2, c-Myc, and CREB in rat peripheral blood mononuclear cells (PBMC) after whole body γ-irradiation and to assess its contribution to biodosimetry. For Western blot experiments, male Wistar rats were whole-body irradiated by a single dose of 0, 0.5, 1, 3, and 5 Gy (60 Co, 1 m, 0.7 Gy/min). As a positive control, leukaemic MOLT-4 cells were used. For ELISA experiments, male Wistar rats were whole-body irradiated by a single dose of 0, 1, 2, 3, 4, and 5 Gy (60 Co, 1 m, 0.6 Gy/min). Blood samples were taken 4 h after the irradiation and PBMC were isolated using centrifugation on Histopaque-1077. Expressions of p21, ATF-2, phospho-ATF-2Thr69/71 , c-Myc, phospho-c-MycThr-58/Ser62 , CREB, and phospho-CREBSer133 were measured using Western blot method. Expression of p21 was also quantified using ELISA. We observed increase of p21 expression in rat PBMC 4 h after irradiation. According to ELISA, p21 levels increased 2.0-, 3.1-, 5.5-, 3.0-, and 3.1fold after irradiation by 1, 2, 3, 4, and 5 Gy, respectively. We did not detect any expression or activation of ATF-2, c-Myc and CREB. Protein p21 could be considered as a perspective biodosimetric marker of clinically significant irradiation (≥1 Gy) in vivo in unstimulated PBMC.

• MeSH
• aktivační transkripční faktory * MeSH
• biologické markery krev   MeSH
• časové faktory MeSH
• celotělové ozáření MeSH
• ELISA MeSH
• inhibitor p21 cyklin-dependentní kinasy metabolismus   účinky záření   MeSH
• ionizující záření * MeSH
• mitogenem aktivované proteinkinasy kinas metabolismus   účinky záření   MeSH
• potkani Wistar MeSH
• protein vázající cAMP responzivní element metabolismus   účinky záření   MeSH
• protoonkogenní proteiny c-myc metabolismus   účinky záření   MeSH
• radiometrie statistika a číselné údaje   MeSH
• vztah dávky záření a odpovědi MeSH
• western blotting MeSH
• záření gama MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

Toxoplasma gondii is a common protozoan parasite that infects warm-blooded animals throughout the world, including mice and humans. During infection, both, the parasite and the host, utilize various mechanisms to maximize their own reproductive success. Mice and humans are both the intermediate hosts for Toxoplasma gondii, which forms specialized vacuoles containing reproductive cysts in the formers' tissue. As half of the human population is infected, developing a disease called toxoplasmosis, along with an ever-growing number of couples suffering with idiopathic infertility, it is therefore surprising that there is a lack of research on how Toxoplasma gondii can alter reproductive parameters. In this study, a detailed histometric screening of the testicular function along with the levels of the pituitary luteinizing hormone (LH) were analysed in infected mice. Data on relative testis and epididymis weight, and sperm count were also collected. Based on the results obtained, the level of LH in the urine of Toxoplasma gondii infected mice was lower compared to the control. In direct correlation with the hormone level, testicular function and sperm production was also significantly lower in Toxoplasma gondii positive group using sperm count and histometric analysis as a marker. Not only were the number of leptotene primary spermatocytes and spermatids lowered, but the number of Sertoli cells and the tubule diameter were elevated. In parallel, a pilot epigenetic study on global testicular methylation, and specific methylation of Crem, Creb1 and Hspa1genes essential for successfully ongoing spermatogenesis was performed. Global methylation was elevated in Toxoplasma infected mice, and differences in the DNA methylation of selected genes were detected between the Toxoplasma positive and control group. These findings demonstrate a direct relation between Toxoplasma gondii infection and the decrease of male reproductive fitness in mice, which may contribute to an increase of idiopathic infertility in humans.

• MeSH
• CpG ostrůvky MeSH
• epididymis metabolismus   parazitologie   patologie   MeSH
• epigeneze genetická MeSH
• exprese genu MeSH
• genetická zdatnost genetika   MeSH
• interakce hostitele a parazita MeSH
• lidé MeSH
• luteinizační hormon genetika   metabolismus   MeSH
• metylace DNA MeSH
• modulátor elementu responzivního pro cyklický AMP genetika   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• oligospermie MeSH
• protein vázající cAMP responzivní element genetika   metabolismus   MeSH
• proteiny tepelného šoku HSP70 genetika   metabolismus   MeSH
• semenotvorné kanálky metabolismus   parazitologie   patologie   MeSH
• Sertoliho buňky metabolismus   parazitologie   patologie   MeSH
• spermie metabolismus   patologie   MeSH
• Toxoplasma patogenita   fyziologie   MeSH
• toxoplazmóza zvířat genetika   metabolismus   parazitologie   patologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Most behavioral and physiological processes in living organisms exhibit periodic circadian rhythmicity. In mammals, these rhythms are coordinated by the circadian clock located in the suprachiasmatic nucleus (SCN) of the hypothalamus. In order to precisely synchronize free-running circadian oscillations to the 24h solar cycle, signals from the external environment, primarily the light/dark cycle, must reach the circadian clock within the SCN. A light pulse elevates intracellular Ca(2+) levels, and activates signaling cascades, leading to transcriptional activation of the clock genes mPer1 and mPer2 via phosphorylation of extracellular-signal-regulated kinases 1/2 (ERK1/2) and cyclic AMP-responsive element binding protein (CREB). Glutamate is the primary excitatory transmitter in retinal terminals in the SCN, and NMDA receptors (NMDAR) are the principal glutamate receptors that mediate the effect of light on resetting the circadian clock. Here we show the circadian rhythm in mRNA expression and protein level of the NMDAR 2B subunit (NR2B) in the SCN, with a peak at night. Also, we demonstrate ifenprodil inhibition of glutamate-induced phosphorylation of CREB (pCREB) and ERK1/2 (pERK1/2), and support thus the evidence for NR2B role in activation of signaling cascade involved in photic resetting of the circadian clock.

• MeSH
• antagonisté excitačních aminokyselin farmakologie   MeSH
• fosforylace MeSH
• hybridizace in situ MeSH
• krysa rodu rattus MeSH
• kyselina glutamová metabolismus   MeSH
• MAP kinasový signální systém MeSH
• nucleus suprachiasmaticus účinky léků   metabolismus   MeSH
• piperidiny farmakologie   MeSH
• potkani Wistar MeSH
• protein vázající cAMP responzivní element metabolismus   MeSH
• receptory N-methyl-D-aspartátu metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The recently discovered prolactin-releasing peptide (PrRP) binds to the PrRP receptor and is involved in endocrine regulation and energy metabolism. However, its main physiological role is currently unknown. Two biologically active isoforms of PrRP exist: the 31 (PrRP31) and the 20 (PrRP20) amino acid forms, which both contain a C-terminal Phe amide sequence. In the present study, the PrRP receptor was immunodetected in three rodent tumor pituitary cell lines: GH3, AtT20 and RC-4B/C cells. The saturation binding of radioiodinated PrRP31 to intact cells demonstrated a K(d) in the 10(-9)M range and a B(max) in the range of tens of thousands binding sites per cell. For binding to RC-4B/C cells, both PrRP31 and PrRP20 competed with (125)I-PrRP31 with a similar K(i). The C-terminal analog PrRP13 showed lower binding potency compared to PrRP31 and PrRP20. All PrRP analogs increased the phosphorylation of MAPK/ERK1/2 (mitogen-activated phosphorylase/extracellular-regulated kinase) and CREB (cAMP response element-binding protein) in RC-4B/C cells. Additionally, prolactin release was induced by the PrRP analogs in a dose-dependent manner in RC-4B/C cells. Finally, food intake after intracerebroventricular administration of PrRP analogs in fasted mice was followed. Both PrRP31 and PrRP20 decreased food intake, but PrRP13 did not show significant effect. Studies on pituitary cell lines expressing the PrRP receptor are more physiologically relevant than those on cells transfected with the receptor. This cell type can be used as a model system for pharmacological studies searching for PrRP antagonists and stable effective PrRP agonists, as these drugs may have potential as anti-obesity agents.

• MeSH
• fosforylace MeSH
• hormon uvolňující prolaktin chemie   metabolismus   MeSH
• hypofyzární hormony sekrece   MeSH
• molekulární sekvence - údaje MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• protein vázající cAMP responzivní element metabolismus   MeSH
• receptory buněčného povrchu metabolismus   MeSH
• sekvence aminokyselin MeSH
• signální transdukce MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH