Phosphofurin acidic cluster sorting protein 2 (PACS2) plays a vital role in maintaining cellular homeostasis by regulating protein trafficking between cellular membranes. This function impacts crucial processes like apoptosis, mitochondria-endoplasmic reticulum interaction, and subsequently Ca2+ flux, lipid biosynthesis, and autophagy. Missense mutations, particularly E209K and E211K, are linked to developmental and epileptic encephalopathy-66 (DEE66), known as PACS2 syndrome. Individuals with this syndrome exhibit neurodevelopmental delay, seizures, facial dysmorphism, hypotonia, and delayed motor skills.Understanding the impact of these missense mutations on molecular processes is crucial. Studies suggest that E209K mutation decreases phosphorylation, increases the survival time of protein, and modifies protein-protein interaction, consequently leading to disruption of calcium flux and lower resistance to apoptosis induction. Unfortunately, to date, only a limited number of research groups have investigated the effects of mutations in the PACS2 gene. Current research on PACS2 syndrome is hampered by the lack of suitable models. While in vitro models using transfected cell lines offer insights, they cannot fully capture the disease's complexity.To address this, utilizing cells from individuals with PACS2 syndrome, specifically induced pluripotent stem cells (iPSCs), holds promise for understanding phenotypic diversity and developing personalized therapies. However, iPSC models may not fully capture tissue-specific effects of the E209K/E211K mutation. In vivo studies using animal models, particularly mice, could overcome these limitations.This review summarizes current knowledge about PACS2 structure and functions, explores the cellular consequences of E209K and E211K mutations, and highlights the potential of iPSC and mouse models in advancing our understanding of PACS2 syndrome.

• MeSH
• indukované pluripotentní kmenové buňky metabolismus   MeSH
• lidé MeSH
• missense mutace * MeSH
• mutace MeSH
• vezikulární transportní proteiny * genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

OBJECTIVE: Transgenic mice with fluorescent protein (FP) reporters take full advantage of new in vivo imaging technologies. Therefore, we generated a TRPC5- and a TRPA1-reporter mouse based on FP C-terminal fusion, providing us with better alternatives for studying the physiology, interaction and coeffectors of these two TRP channels at the cellular and tissue level. METHODS: We generated transgenic constructs of the murine TRPC5- and TRPA1-gene with a 3*GGGGS linker and C-terminal fusion to mCherry and mTagBFP, respectively. We microinjected zygotes to generate reporter mice. Reporter mice were examined for visible fluorescence in trigeminal ganglia with two-photon microscopy, immunohistochemistry and calcium imaging. RESULTS: Both TRPC5-mCherry and TRPA1-mTagBFP knock-in mouse models were successful at the DNA and RNA level. However, at the protein level, TRPC5 resulted in no mCherry fluorescence. In contrast, sensory neurons derived from the TRPA1-reporter mice exhibited visible mTag-BFP fluorescence, although TRPA1 had apparently lost its ion channel function. CONCLUSIONS: Creating transgenic mice with a TRP channel tagged at the C-terminus with a FP requires detailed investigation of the structural and functional consequences in a given cellular context and fine-tuning the design of specific constructs for a given TRP channel subtype. Different degrees of functional impairment of TRPA1 and TRPC5 constructs suggest a specific importance of the distal C-terminus for the regulation of these two channels in trigeminal neurons.

• MeSH
• červený fluorescenční protein MeSH
• ganglion trigeminale metabolismus   MeSH
• genový knockin * MeSH
• kationtové kanály TRPC * genetika   metabolismus   MeSH
• kationtový kanál TRPA1 * genetika   metabolismus   MeSH
• luminescentní proteiny * genetika   metabolismus   MeSH
• myši transgenní * MeSH
• myši MeSH
• rekombinantní fúzní proteiny metabolismus   genetika   MeSH
• vápník metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

A tubular microdialysis probe is made from polysulfone hollow fibre for human haemodialysis, which has an inner diameter of 200 μm and a thickness of 20 μm. Milk is deposited to the outer surface of the hollow fibre and allowed to dry to form a dry sample. The tubular probe is then connected to the syringe pump and microdialysis of the dry sample into 0.5 mol/L HCl as acceptor is performed. 2.5 μL of microdialysate is obtained and analyzed for inorganic cations by capillary electrophoresis with contactless conductivity detection. Baseline separation of NH4+, K+, Ca2+, Na+, Mg2+, Li+ is achieved in 5.5 mol/L acetic acid as background electrolyte using a fused silica capillary with inner diameter of 25 μm and length of 31.5 cm. The reproducibility of dry sample microdialysis including CE analysis for peak area ranges from 2.4 to 3.9 % after normalization to Li+ as internal standard.

• MeSH
• elektroforéza kapilární * přístrojové vybavení   metody   MeSH
• kationty * analýza   MeSH
• mikrodialýza * přístrojové vybavení   metody   MeSH
• mléko * chemie   MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH

OBJECTIVES: To compare the drug survival of etanercept to monoclonal tumour necrosis factor-α inhibitors in rheumatoid arthritis, ankylosing spondylitis and psoriatic arthritis. METHODS: Patients initiating first line biological therapy with tumour necrosis factor-α were propensity score matched and compared for drug survival with a Kaplan-Meier analysis. RESULTS: We matched 657 to 657 patients in rheumatoid arthritis, the median survival time on etanercept was 44.6 months vs. 36.8 months on monoclonal antibody tumour necrosis factor-α inhibitors, with a hazard ratio of 0.94, p = 0.416 We matched 187 to 356 patients in ankylosing spondylitis, the median survival time on etanercept was 75.1 compared to 68.0 months, hazard ratio of 0.78, p = 0.087 We matched 81 to 160 psoriatic arthritis patients, the median survival time on etanercept was 35.8. compared to 65.7 months, hazard ratio 1.61, p = 0.011. Patients treated with etanercept had significantly worse psoriasis scoring during follow up. CONCLUSIONS: We found comparable survival in rheumatoid arthritis and ankylosing spondylitis. In psoriatic arthritis, we found significantly shorter survival on etanercept, possibly due to worse response of skin and nail manifestations.

• MeSH
• adalimumab terapeutické užití   MeSH
• ankylózující spondylitida * farmakoterapie   mortalita   MeSH
• antirevmatika * terapeutické užití   MeSH
• dospělí MeSH
• etanercept * terapeutické užití   MeSH
• infliximab terapeutické užití   MeSH
• Kaplanův-Meierův odhad MeSH
• lidé středního věku MeSH
• lidé MeSH
• monoklonální protilátky terapeutické užití   MeSH
• psoriatická artritida * farmakoterapie   mortalita   MeSH
• registrace * MeSH
• revmatoidní artritida * farmakoterapie   mortalita   MeSH
• senioři MeSH
• tendenční skóre * MeSH
• TNF-alfa * antagonisté a inhibitory   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Geografické názvy
• Česká republika MeSH

Vápník a fosfor jsou důležitými minerály, na jejichž metabolismu a homeostáze se v organismu podílí celá řada hormonů a orgánů, zejména skelet, příštítná tělíska, gastrointestinální trakt a ledviny. Poruchy fosfokalciového metabolismu mohou být spojeny s nefyziologickými koncentracemi těchto minerálů v séru, patologickými ději v kostní tkáni a poruchami hlavních regulačních systémů, hlavně příštítných tělísek, ledvin a gastrointestinálního traktu.

Calcium and phosphorus are important minerals whose metabolism and homeostasis in the body involve many hormones and organs, especially the skeleton, parathyroid glands, digestive tract and kidneys. Disturbances in phospho-calcium metabolism may be associated with non-physiological serum concentrations of these minerals, pathological processes in bone tissue, and disturbances in major regulatory systems, particularly the parathyroid glands, kidneys, and gastrointestinal tract.

• MeSH
• fosfor * krev   MeSH
• hyperfosfatemie diagnóza   komplikace   MeSH
• hyperkalcemie diagnóza   farmakoterapie   komplikace   MeSH
• hypofosfatemie diagnóza   komplikace   MeSH
• hypokalcemie diagnóza   farmakoterapie   komplikace   MeSH
• kalcitriol krev   metabolismus   MeSH
• lidé MeSH
• metabolické nemoci diagnóza   farmakoterapie   komplikace   metabolismus   MeSH
• nedostatek vitaminu D diagnóza   farmakoterapie   krev   MeSH
• vápník * krev   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Nedd4-2 E3 ligase regulates Na+ homeostasis by ubiquitinating various channels and membrane transporters, including the epithelial sodium channel ENaC. In turn, Nedd4-2 dysregulation leads to various conditions, including electrolytic imbalance, respiratory distress, hypertension, and kidney diseases. However, Nedd4-2 regulation remains mostly unclear. The present study aims at elucidating Nedd4-2 regulation by structurally characterizing Nedd4-2 and its complexes using several biophysical techniques. Our cryo-EM reconstruction shows that the C2 domain blocks the E2-binding surface of the HECT domain. This blockage, ubiquitin-binding exosite masking by the WW1 domain, catalytic C922 blockage and HECT domain stabilization provide the structural basis for Nedd4-2 autoinhibition. Furthermore, Ca2+-dependent C2 membrane binding disrupts C2/HECT interactions, but not Ca2+ alone, whereas 14-3-3 protein binds to a flexible region of Nedd4-2 containing the WW2 and WW3 domains, thereby inhibiting its catalytic activity and membrane binding. Overall, our data provide key mechanistic insights into Nedd4-2 regulation toward fostering the development of strategies targeting Nedd4-2 function.

• MeSH
• elektronová kryomikroskopie MeSH
• HEK293 buňky MeSH
• lidé MeSH
• molekulární modely MeSH
• proteinové domény MeSH
• proteiny 14-3-3 * metabolismus   chemie   MeSH
• ubikvitinace MeSH
• ubikvitinligasy Nedd4 * metabolismus   chemie   genetika   ultrastruktura   MeSH
• vápník * metabolismus   MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Mitochondria are key to cellular energetics, metabolism, and signaling. Their dysfunction is linked to devastating diseases, including mitochondrial disorders, diabetes, neurodegenerative diseases, cardiac disorders, and cancer. Here, we present a knockout mouse model lacking the complex IV assembly factor SMIM20/MITRAC7. SMIM20-/- mice display cardiac pathology with reduced heart weight and cardiac output. Heart mitochondria present with reduced levels of complex IV associated with increased complex I activity, have altered fatty acid oxidation, and display elevated levels of ROS production. Interestingly, mutant mouse ventricular myocytes show unphysiological Ca2+ handling, which can be attributed to the increase in mitochondrial ROS production. Our study presents an example of a tissue-specific phenotype in the context of OXPHOS dysfunction. Moreover, our data suggest a link between complex IV dysfunction and Ca2+ handling at the endoplasmic reticulum through ROS signaling.

• MeSH
• endoplazmatické retikulum metabolismus   MeSH
• kardiomyocyty metabolismus   MeSH
• membránové proteiny * metabolismus   genetika   MeSH
• mitochondriální proteiny * metabolismus   genetika   MeSH
• myokard * metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• oxidativní fosforylace MeSH
• proteiny dánia pruhovaného MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• respirační komplex IV * metabolismus   MeSH
• srdeční mitochondrie metabolismus   MeSH
• vápník metabolismus   MeSH
• vápníková signalizace * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: This study aimed to evaluate the effect of overweight and obesity at the start of anti-TNF therapy on treatment response and relapse rate in children with inflammatory bowel disease (IBD). METHODS: This multicenter, retrospective cohort study included 22 IBD centers in 14 countries. Children diagnosed with IBD in whom antitumor necrosis factor (anti-TNF) was introduced were included; those who were overweight/obese were compared with children who were well/undernourished. RESULTS: Six hundred thirty-seven children (370 [58%] males; mean age 11.5 ± 3.5 years) were included; 140 (22%) were in the overweight/obese group (OG) and 497 (78%) had BMI ≤1 SD (CG). The mean follow-up time was 141 ± 78 weeks (median 117 weeks). There was no difference in the loss of response (LOR) to anti-TNF between groups throughout the follow-up. However, children in OG had more dose escalations than controls. Male sex and lack of concomitant immunomodulators at the start of anti-TNF were risk factors associated with the LOR. There was no difference in the relapse rate in the first year after anti-TNF introduction; however, at the end of the follow-up, the relapse rate was significantly higher in the OG compared with CG (89 [64%] vs 218 [44%], respectively, P < .001). Univariate and multivariate analysis revealed that being overweight/obese, having UC, or being of male sex were factors associated with a higher risk for relapse. CONCLUSIONS: Overweight/obese children with IBD were not at a higher risk of LOR to anti-TNF. Relapse in the first year after anti-TNF was introduced, but risk for relapse was increased at the end of follow-up.

• MeSH
• dítě MeSH
• idiopatické střevní záněty * farmakoterapie   komplikace   MeSH
• infliximab terapeutické užití   MeSH
• lidé MeSH
• mladiství MeSH
• nadváha * komplikace   MeSH
• následné studie MeSH
• obezita * komplikace   MeSH
• recidiva MeSH
• retrospektivní studie MeSH
• rizikové faktory MeSH
• TNF-alfa * antagonisté a inhibitory   MeSH
• výsledek terapie MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH

Výživa hraje významnou roli v léčbě chronického onemocnění ledvin (CHOL). Jedním z hlavních cílů nutriční terapie CHOL je umožnění správného vývoje a růstu dětí. Vhodná výživa je zásadní pro terapii komplikací CHOL, obzvláště metabolické kostní nemoci. Zdravá, čerstvá a pestrá strava by měla být základem jídelníčku, příjem zpracovaných potravin by měly děti s CHOL významně omezit. Tento přehledový článek vychází z recentně publikovaných mezinárodních doporučení a shrnuje současný přístup k výživě dětí s CHOL.

Nutrition plays an important role in the management of chronic kidney disease (CKD). One of the main nutritional goals in children is to achieve normal growth and develop­ment. Adequate nutrition is crucial for proper management of CKD complications, especially metabolic bone disease. A healthy, fresh, and varied diet should form the basis of the meal plan, patients should significantly limit their intake of processed foods. This review is based on recently published international guidelines and summarizes the current approach to nutrition of children with CKD.

• MeSH
• chronická renální insuficience * diagnóza   MeSH
• dítě MeSH
• draslík krev   MeSH
• fosfor krev   MeSH
• hodnocení stavu výživy MeSH
• kojenec MeSH
• lidé MeSH
• nutriční terapie metody   MeSH
• předškolní dítě MeSH
• strava, jídlo, výživa MeSH
• vápník krev   MeSH
• výživové doporučené dávky MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• předškolní dítě MeSH
• Publikační typ
• přehledy MeSH

The sodium/calcium exchanger (NCX) type 1 has been well described in various cancers, but little is known about the other two NCX types (NCX2 and NCX3). In this study, we used the selective blocker of NCX3 - YM-244769 to investigate changes in apoptosis induction, migration, proliferation, intracellular calcium and ATP in four cancer cell lines - DLD1, HeLa, MDA-MB-231 and JIMT1. In all four cell lines we observed a concentration-dependent increase in the number of apoptotic cells, as well as reduced migration and proliferation. Induction of hypoxic conditions did not alter the response of these cells to YM-244769 in any of the above-mentioned parameters. These results indicate the role of NCX3 in cancer cell migration, proliferation and apoptosis, as inhibition of NCX1 by the specific blocker SEA0400 had no significant effect on these parameters. However, we verified the effect of NCX3 inhibition by using CRISPR/Cas9 to generate clones in which the SLC8A3 (NCX3) gene was deleted, and we obtained the same results. In addition, mitochondrial respiration was impaired in the clones with NCX3 knocked-out, suggesting that NCX3 also play a role in bioenergetics. In conclusion, we have clearly shown that NCX3 plays an important anti-apoptotic, pro-migratory and proliferative role in the cancer cells by affecting mitochondrial bioenergetics, thus supporting their survival and fate.

• MeSH
• apoptóza účinky léků   MeSH
• lidé MeSH
• mitochondrie metabolismus   účinky léků   MeSH
• nádorové buněčné linie MeSH
• nádory * metabolismus   patologie   genetika   MeSH
• pohyb buněk účinky léků   MeSH
• proliferace buněk účinky léků   MeSH
• pumpa pro výměnu sodíku a vápníku * metabolismus   genetika   antagonisté a inhibitory   MeSH
• vápník metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH