The aim of the present study was to assess systemic circulatory and tissue activities of both the classical arm and of the alternative arm of the renin-angiotensin system (RAS) in a new transgenic rat line (TG7371) that expresses angiotensin-(1-7) (ANG 1-7)-producing fusion protein; the results were compared with the activities measured in control transgene-negative Hannover Sprague-Dawley (HanSD) rats. Plasma and tissue concentrations of angiotensin II (ANG II) and ANG 1-7, and kidney mRNA expressions of receptors responsible for biological actions of ANG II and ANG 1-7 [i.e. ANG II type 1 and type 2 (AT1 and AT2) and Mas receptors] were assessed in TG7371 transgene-positive and in HanSD rats. We found that male TG7371 transgene-positive rats exhibited significantly elevated plasma, kidney, heart and lung ANG 1-7 concentrations as compared with control male HanSD rats; by contrast, there was no significant difference in ANG II concentrations and no significant differences in mRNA expression of AT1, AT2 and Mas receptors. In addition, we found that in male TG7371 transgene-positive rats blood pressure was lower than in male HanSD rats. These data indicate that the balance between the classical arm and the alternative arm of the RAS was in male TGR7371 transgene-positive rats markedly shifted in favor of the latter. In conclusion, TG7371 transgene-positive rats represent a new powerful tool to study the long-term role of the alternative arm of the RAS in the pathophysiology and potentially in the treatment of cardio-renal diseases.

• MeSH
• angiotensin I * metabolismus   MeSH
• angiotensin II * MeSH
• kardiovaskulární nemoci metabolismus   genetika   MeSH
• krevní tlak fyziologie   MeSH
• krysa rodu rattus MeSH
• ledviny metabolismus   MeSH
• nemoci ledvin metabolismus   genetika   MeSH
• peptidové fragmenty * metabolismus   MeSH
• potkani Sprague-Dawley * MeSH
• potkani transgenní * MeSH
• protoonkogen Mas MeSH
• receptor angiotensinu typ 1 genetika   metabolismus   MeSH
• receptory spřažené s G-proteiny genetika   metabolismus   MeSH
• rekombinantní fúzní proteiny metabolismus   MeSH
• renin-angiotensin systém * fyziologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Spondyloartritidy jsou heterogenní skupina chronických zánětlivých onemocnění s pestrou škálou projevů, z nichž dominuje postižení axiálního skeletu, periferních kloubů, entezitida a daktylitida, časté jsou též projevy extraskeletální. Lze je dělit podle dominantního postižení na axiální či periferní spondyloartritidy, axiální pak dále podle radiograficky prokazatelných strukturálních změn na sakroiliakálních skloubeních na radiografickou a nonradiografickou formu. Základem léčby nadále zůstávají nesteroidní antirevmatika, významný pokrok však nastal zejména se zavedením biologické léčby, kterou je v současnosti dle mezinárodních doporučení možno nasadit již v druhé linii léčby. Jednou ze základních skupin biologických léčiv jsou inhibitory tumor nekrotizujícího faktoru a, jejichž zástupcem je i certolizumab pegol, humanizovaná monoklonální protilátka, která na rozdíl od ostatních zástupců této skupiny obsahuje pouze Fab fragment molekuly s dvěma navázanými molekulami polyethylenglykolu. Tato struktura minimalizuje transplacentární přestup, a proto je certolizumab pegol s výhodou užíván po celou dobu těhotenství u těhotných pacientek s revmatickým onemocněním. V následujícím textu demonstrujeme účinnost i bezpečnost užití certolizumab pegolu u dvou těhotných pacientek s axiální spondyloartritidou.

Spondyloarthritis is a heterogeneous group of chronic inflammatory diseases with various symptoms that mainly include the damage of axial skeleton, peripheral joints, enthesitis and dactylitis. Extraskeletal manifestations are often as well. Spondyloarthritis can be divided into two groups, axial and peripheral, depending on the dominant involvement. Axial type can be further divided into radiographic and non-radiographic type based on the radiographic evidence of structural damage of sacroiliac joints. Nonsteroidal anti-inflammatory drugs still remain the drug of the first choice, but it was the introduction of biological therapy that led to a significant treatment progress. According to the most recent international recommendations, biological therapy is approved as a second-line treatment of the disease. Tumor necrosis factor a inhibitors are one of the main groups of biologics that include certolizumab pegol, a humanized monoclonal antibody that consists of a single Fab fragment with no Fc portion compared to the other group agents. The Fab fragment is conjugated with two molecules of polyethylene glycol which minimizes the placental transfer. Therefore, certolizumab pegol is successfully used throughout pregnancy in pregnant women with rheumatic diseases. The following text demonstrates both the efficacy and safety of use of certolizumab pegol in two pregnant patients treating for axial spondyloarthritis.

Recurrent epidemics and pandemics caused by seasonal human influenza viruses result in substantial morbidity and are a significant public health burden worldwide annually. Antiviral drugs are used to treat influenza infections but have several limitations.. Therefore, monoclonal antibody therapy is an exciting and promising approach. Nanobodies, also known as single-domain antibodies, are a new class derived from heavy-chain-only antibodies found in camelids like alpacas, llamas, and camels. These antibodies neutralize influenza viruses by targeting various proteins through multiple mechanisms. For example, they can target the hemagglutinin protein to prevent its functions. By focusing on conserved epitopes, they can neutralize a variety of influenza subtypes, including seasonal flu strains and possible pandemic variants. Additionally, these antibodies can neutralize free-floating viruses in the extracellular environment, preventing them from infecting cells. They can reduce the viral load and limit the spread of the infection. Using nanobodies to neutralize influenza viruses provides numerous advantages compared to conventional antibodies. Thanks to their unique properties, nanobodies play a crucial role in fighting influenza, improving disease management, and strengthening public health responses. In this review, we summarize the role of nanobodies in influenza virus neutralization.

• MeSH
• chřipka lidská * imunologie   virologie   MeSH
• jednodoménové protilátky * imunologie   terapeutické užití   MeSH
• lidé MeSH
• neutralizující protilátky * imunologie   terapeutické užití   MeSH
• Orthomyxoviridae * imunologie   MeSH
• protilátky virové * imunologie   terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

INTRODUCTION: Plasma phosphorylated tau (p-tau)181, glial fibrillary acidic protein (GFAP), neurofilament light chain (NfL), and amyloid beta ratio (Aβ42/40) may have diagnostic and prognostic value in Alzheimer's disease (AD). Here we assess which markers can best identify AD from controls and other non-AD dementias in a large international multi-center study. METHODS: Plasma samples (n = 1298) were collected from six international centers. Aβ40, Aβ42, GFAP, NfL, and p-tau181 were measured using single molecule array. In each group, AD diagnosis/co-pathology was defined according to cerebrospinal fluid biomarkers or amyloid positron emission tomography. Validations were performed in three separate cohorts via single and dual cut-off models. RESULTS: p-tau181 showed the best area under the curve value to separate AD from frontotemporal dementia, controls, and Aβ- dementia with Lewy bodies. However, this discriminative power could not be reproduced by applying pre-defined cut-offs. DISCUSSION: p-tau181 was the best single plasma marker for detecting AD at any stage. Specific cut-offs are needed to maximize diagnostic performances. HIGHLIGHTS: Phosphorylated tau (p-tau)181 provided a clear differentiation between controls and Alzheimer's disease (AD) participants, with evidence of increased levels in the preclinical stage of AD. Plasma biomarkers demonstrated that when amyloid co-pathology is removed from dementia with Lewy bodies (DLB), only glial fibrillary acidic protein and neurofilament light chain remain to predict DLB. Given the low prevalence of amyloid co-pathology in frontotemporal dementia (FTD), p-tau181 and its ratio with amyloid beta 42 are strong biomarkers to differentiate FTD from AD.

• MeSH
• Alzheimerova nemoc * krev   diagnóza   MeSH
• amyloidní beta-protein * krev   MeSH
• biologické markery krev   MeSH
• demence * krev   diagnóza   MeSH
• fosforylace MeSH
• gliový fibrilární kyselý protein * krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• neurofilamentové proteiny * krev   MeSH
• peptidové fragmenty * krev   MeSH
• proteiny tau * krev   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH

One of the main challenges in analyzing chemical messengers in the brain is the optimization of tissue sampling and preparation protocols. Limiting postmortem time and terminating enzyme activity is critical to identify low-abundance neurotransmitters and neuropeptides. Here, we used a rapid and uniform conductive heat transfer stabilization method that was compared with a conventional fresh freezing protocol. Together with a selective chemical derivatization method and an optimized quantitation approach using deuterated internal standards, we spatially mapped neurotransmitters and their related metabolites by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) in rat brain tissue sections. Although the heat stabilization did not show differences in the levels of dopamine, norepinephrine, and serotonin, their related metabolites 3,4-dihydroxyphenylacetaldehyde, 3,4-dihydroxyphenylacetic acid, homovanillic acid, 3-methoxy-4-hydroxyphenylacetaldehyde, dihydroxyphenylethyleneglycol, and 5-hydroxyindoleacetic acid were all significantly lower, indicating reduced neurotransmitter postmortem turnover ratios. Heat stabilization enabled detection of an increased number and higher levels of prodynorphin, proenkephalin, and tachykinin-derived bioactive neuropeptides. The low-abundant C-terminal flanking peptide, neuropeptide-γ, and nociceptin remained intact and were exclusively imaged in heat-stabilized brains. Without heat stabilization, degradation fragments of full-length peptides occurred in the fresh frozen tissues. The sample preparation protocols were furthermore tested on rat brains affected by acute anesthesia induced by isoflurane and medetomidine, showing comparable results to non-anesthetized animals on the neurotransmitters level without significant changes. Our data provide evidence for the potential use of heat stabilization prior to MALDI-MSI analyses to improve the examination of the in vivo state of neuronal chemical messengers in brain tissues not impacted by prior acute anesthesia.

• MeSH
• krysa rodu rattus MeSH
• mozek - chemie * fyziologie   MeSH
• mozek * metabolismus   MeSH
• neurony * metabolismus   chemie   MeSH
• neurotransmiterové látky * metabolismus   analýza   MeSH
• potkani Sprague-Dawley MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice * metody   MeSH
• vysoká teplota * MeSH
• zmrazování MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

3-methylglutaconic aciduria (3-MGCA) is a biochemical finding in a diverse group of inherited metabolic disorders. Conditions manifesting 3-MGCA are classified into two major categories, primary and secondary. Primary 3-MGCAs involve two inherited enzymatic deficiencies affecting leucine catabolism, whereas secondary 3-MGCAs comprise a larger heterogeneous group of conditions that have in common compromised mitochondrial energy metabolism. Here, we report 3-MGCA in two siblings presenting with sensorineural hearing loss and neurological abnormalities associated with a novel, homozygous missense variant (c.1999C>G, p.Leu667Val) in the YME1L1 gene which encodes a mitochondrial ATP-dependent metalloprotease. We show that the identified variant results in compromised YME1L1 function, as evidenced by abnormal proteolytic processing of substrate proteins, such as OPA1 and PRELID1. Consistent with the aberrant processing of the mitochondrial fusion protein OPA1, we demonstrate enhanced mitochondrial fission and fragmentation of the mitochondrial network in patient-derived fibroblasts. Furthermore, our results indicate that YME1L1L667V is associated with attenuated activity of rate-limiting Krebs cycle enzymes and reduced mitochondrial respiration, which may explain the build-up of 3-methylglutaconic and 3-methylglutaric acid due to the diversion of acetyl-CoA, not efficiently processed in the Krebs cycle, towards the formation of 3-methylglutaconyl-CoA, the precursor of these metabolites. In summary, our findings classify YME1L1 deficiency as a new type of secondary 3-MGCA, thus expanding the genetic landscape and facilitating the diagnosis of inherited metabolic disorders featuring this biochemical phenotype.

• MeSH
• dítě MeSH
• fibroblasty metabolismus   MeSH
• glutaráty MeSH
• lidé MeSH
• metaloendopeptidasy * genetika   metabolismus   MeSH
• missense mutace MeSH
• mitochondriální dynamika MeSH
• mitochondriální proteiny * genetika   MeSH
• mitochondrie metabolismus   MeSH
• percepční nedoslýchavost genetika   MeSH
• sourozenci MeSH
• vrozené poruchy metabolismu * genetika   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH

Male infertility is a multifactorial condition contributing to approximately 50% of all cases of couple infertility. In recent years, significant advances have been made in both diagnostics and treatment. This review summarizes key developments from 2019 to 2024 with direct relevance to routine clinical practice in Czech urology and andrology. Particular attention is paid to the updated semen analysis standards (World Health Organisation 6th edition, 2021), sperm DNA fragmentation testing, genetic evaluation (karyotyping, Y chromosome microdeletions, and exome sequencing), surgical management of varicocele, and sperm retrieval techniques for azoospermia, including microdissection testicular sperm extraction (micro-TESE). The article also discusses pharmacological options (gonadotropins, selective estrogen receptor modulators, antioxidants), the impact of lifestyle factors, and the importance of interdisciplinary collaboration with assisted reproduction centers. Future perspectives, including the role of preventive strategies in male reproductive health, are also addressed. The aim is to provide a comprehensive and clinically applicable overview of current recommendations and therapeutic approaches in andrology, with a focus on their implementation in the Czech urological setting.

• MeSH
• analýza spermatu metody   MeSH
• antioxidancia farmakologie   terapeutické užití   MeSH
• asistovaná reprodukce MeSH
• genetické testování metody   MeSH
• gonadotropiny terapeutické užití   MeSH
• lidé MeSH
• mužská infertilita * diagnóza   etiologie   terapie   MeSH
• odběr spermií MeSH
• selektivní modulátory estrogenních receptorů farmakologie   terapeutické užití   MeSH
• varikokéla chirurgie   MeSH
• životní styl MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• systematický přehled MeSH

Cíl: Vztah mezi hypercholesterolemií, zvláště zvýšenými hodnotami cholesterolu v lipoproteinech o nízké hustotě (lDl-c) a ischemickou chorobou srdeční (ichS), je potvrzen důkazy z předchozích epidemiologic kých studií. Důležité bylo popsání souvislosti mezi genetickým polymorfismem a hodnotami lipidů v plazmě. cílem této studie bylo zkoumat vztah mezi variantami rs5918 t>c genu pro itGB3 a variantou rs1799837 c>t ge nu pro APo-A1 na jedné straně a metabolismem lipidů v séru na straně druhé. Pacienti a metody: Do této studie bylo zařazeno celkem 100 jedinců s ichS a 250 zdravých jedinců. u každé ho účastníka bylo provedeno základní biochemické vyšetření včetně stanovení koncentrace glukózy v séru, celkového cholesterolu (total cholesterol, tc) v séru, hodnot hDl-c, lDl-c a triglyceridů. Pro genotypizaci polymorfismů genů pro itGB3 a APo-A1 byla použita polymerázová řetězová reakce s následnou analýzou polymorfismu délky restrikčních fragmentů (restriction fragment length polymorphism, rflP). Výsledky: Pokud se týče genotypu a distribuce alel polymorfismu rs5918 t>c genu pro itGB3, vyskytovala se alela c častěji ve skupině s ichS než v kontrolní skupině (p = 0,001). Ve skupině s ichS byla navíc pozorována statisticky významná souvislost mezi genotypem rs5918 itGB3 a hodnotami celkového cholesterolu v geno typu cc na jedné straně a tc v séru a cholesterolu v lipoproteinech o vysoké hustotě (hDl-c) (p = 0,0006, resp. p = 0,016). nicméně ani v kontrolní skupině, ani ve skupině s ichS nebyla nalezena statisticky významná spojitost mezi polymorfismem rs1799837 c>t genu pro APo-A1 a biochemickými parametry. Závěr: Výsledky prokázaly, že varianta rs5918 t>c genu pro itGB3 by mohla být z klinického hlediska vý znamná jako genetický marker zvýšené vnímavosti k rozvoji ichS. Alelu c varianty rs5918 genu pro itGB3 by tak bylo možno navrhnout pro screening potenciálního rozvoje ichS u populace kyperských turků, kteří se dostaví na kontrolu k lékaři.

Aim: The relationship between hypercholesterolemia, particularly elevated low density lipoprotein-cholesterol (LDL-C) levels and coronary artery disease is recognized by the evidence from previous epidemiologic studies. Importantly, genetic polymorphisms on different genes have been reported to be associated with plasma lipid levels. In this particular study, we aimed to investigate the relationship between the ITGB3 gene rs5918 T>C and APO-A1 gene rs1799837 C>T markers and serum lipid metabolism. Patients and methods: A total of 100 subjects with CAD and 250 healthy subjects were involved in the current study. A basic biochemical analysis, including serum glucose, total serum cholesterol, HDL-C, LDL-C and triglycerides, was performed for each participant. Genotyping for the ITGB3 gene and APO-A1 gene polymorphisms was performed by polymerase chain reaction followed by restriction fragment length polymorphism (RFLP) analysis. Results: With respect to the genotype and allele distributions of ITGB3 rs5918 T>C polymorphism, the frequency of the C allele was higher in the coronary artery disease (CAD) group compared to the control group (p = 0.001). Moreover, there was a statistically significant association detected between ITGB3 rs5918 CC genotype and serum total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) (p = 0.0006, p = 0.016, respectively) in CAD group. However there was no statistically significant association was identified between the APOA1 rs1799837 C>T polymorphism and biochemical parameters in control and CAD group. Conclusion: The results demonstrated that rs5918 T>C variant within the ITGB3 gene might have a clinical importance as a genetic marker which increases the susceptibility to CAD. Therefore, the ITGB3 gene rs5918 C allele may be offered as a screening option for CAD in Turkish Cypriot population who come in for medical check-up.

BACKGROUND: The treatment of non-small cell lung cancer (NSCLC) patients is correlated with the efficacy of immune checkpoint blockade therapy (ICB) targeting programmed cell death ligand 1 (PD-L1) or its cognate receptor (PD-1) on cancer cells or infiltrating immune cells. Analysis of PD-L1/PD-1 expression in tumor tissue represents a crucial step before PD-L1/PD-1 blocker usage. METHODS: We used directed evolution of protein variants derived from a 13 kDa Myomedin loop-type combinatorial library with 12 randomized amino acid residues to select high-affinity binders of human PD-L1 (hPD-L1). After the ribosome display, individual clones were screened by ELISA. Detailed analysis of binding affinity and kinetics was performed using LigandTracer. The specificity of Myomedins was assessed using fluorescent microscopy on HEK293T-transfected cells and cultured cancer cells in vitro, formalin-fixed paraffin-embedded (FFPE) sections of human tonsils, and FFPE tumor samples of NSCLC patients. RESULTS: Seven identified PD-L1 binders, called MLE, showed positive staining for hPD-L1 on transfected HEK293T cells and cultured MCF-7 cells. MLE031, MLE105, MLE249, and MLE309 exhibited high affinity to both human and mouse PD-L1-transfected HEK293T cells measured with LigandTracer. The diagnostic potential of MLE variants was tested on human tonsillitis tissue and compared with diagnostic anti-PD-L1 antibody DAKO 28-8 and PD-L1 IHC 22C3 pharmDx antibody. MLE249 and MLE309 exhibited an excellent overlap with diagnostic DAKO 28-8 (Pearson ́s coefficient (r) = 0.836 and 0.731, respectively) on human tonsils on which MLE309 exhibited also excellent overlap with diagnostic 22C3 antibody (r = 0.876). Using three NSCLC tissues, MLE249 staining overlaps with 28-8 antibody (r = 0.455-0.883), and MLE309 exhibited overlap with 22C3 antibody (r = 0.534-0.619). Three MLE proteins fused with Fc fragments of rabbit IgG, MLE249-rFc, MLE309-rFc and MLE031-rFc, exhibited very good overlap with anti-PD-L1 antibody 28-8 on tonsil tissue (r = 0.691, 0.610, and 0.667, respectively). Finally, MLE249-rFc, MLE309-rFc and MLE031-rFc exhibited higher sensitivity in comparison to IHC 22C3 antibody using routine immunohistochemistry staining system Ventana, which is one of gold standards for PD-L1 diagnosis. CONCLUSIONS: We demonstrated the development of MLE Myomedins specifically recognizing hPD-L1 that may serve as a refinement tool for clinical PD-L1 detection.

• MeSH
• antigeny CD274 * metabolismus   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory plic * diagnóza   metabolismus   patologie   MeSH
• nemalobuněčný karcinom plic * diagnóza   metabolismus   patologie   MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The use of microfluidic sperm sorting (MFSS) systems in infertility treatment is increasing due to their practicality and ease of use. While often presented as highly effective, their efficacy in patients with varying sperm analysis results remains uncertain. In this study, we evaluated the effectiveness of MFSS compared with the swim-up (SU) technique in terms of oxygen radical levels and spermiogram parameters. Samples from each patient were processed using both methods, followed by assessments of sperm concentration, motility, morphology, DNA integrity, acrosomal status, and mitochondrial membrane potential. Participants were selected based on sperm analysis and categorized as normozoospermic (n = 40) or non-normozoospermic (n = 28). An analysis of separation techniques revealed no significant differences, except for a lower percentage of DNA-fragmented sperm in the MFSS group compared with SU within the non-normozoospermic cohort (SU: 10.0% vs. MFSS: 5.69%, p = 0.027). No differences were observed between SU and MFSS in normozoospermic men. The MFSS method is a simple technique, frequently used in laboratories, that yields good results but does not offer a substantial advantage over SU. The primary benefit of MFSS appears to be a significant reduction in the proportion of sperm with DNA fragmentation compared with SU in patients with abnormal sperm analysis results.

• MeSH
• analýza spermatu metody   MeSH
• dospělí MeSH
• fragmentace DNA MeSH
• intracytoplazmatické injekce spermie * metody   MeSH
• lidé MeSH
• membránový potenciál mitochondrií MeSH
• mikrofluidika * metody   MeSH
• motilita spermií * MeSH
• mužská infertilita terapie   MeSH
• separace buněk * metody   MeSH
• spermie * cytologie   metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH