A huge number of inbred mouse strains with different bone properties have become available for musculoskeletal research. C57Bl/6J and C3H/HeOuJ mice show a significant difference in their bone characteristics. Nevertheless, there is a lack of knowledge on the molecular basis of these strain differences. The aim of this study is to determine the gene expression of selected regulators expressed in the bone marrow as well as bone microstructure of C57Bl/6J and C3H/HeOuJ mice. Bone properties were investigated in 20-week-old female C57Bl/6J and C3H/HeOuJ mice. Total RNA was extracted from the bone marrow of the tibia and gene expression of the following genes was determined by quantitative real-time PCR: SOST, DKK1, OPN, FGF23, RANKL, IL6, TNF, IL17a, and OPG. The femur and third lumbar vertebral body (L3) were investigated by μCT. Bone histomorphometric evaluations were performed in tartrate-resistant acid phosphatase/toluidine blue stained fourth lumbar vertebral bodies (L4). C57Bl/6J and C3H/HeOuJ mice showed significant differences in the gene expression of DKK1, FGF23, IL-6, TNF, and OPG. When compared with C57Bl/6J mice, C3H/HeOuJ mice had a stronger cortical and trabecular bone microstructure at the femur. In contrast, at L3 bone volume/total volume (BV/TV) and trabecular number were significantly higher in C57Bl/6J than in C3H/HeOuJ mice. Bone histomorphometry of L4 revealed significantly higher BV/TV, trabecular number, and thickness in C57Bl/6J mice. Furthermore, the number of osteoblasts and the number of osteoclasts/bone perimeter were higher in the C57Bl/6J mice. This study shows that C57Bl/6J and C3H/HeOuJ mice exhibit a differential expression of cytokines present in the bone marrow. Bone properties differ not only between both strains but also in relation to the investigated bone region.

• Publikační typ
• časopisecké články MeSH

Pathology diagnostics relies on the assessment of morphology by trained experts, which remains subjective and qualitative. Here we developed a framework for large-scale histomorphometry (FLASH) performing deep learning-based semantic segmentation and subsequent large-scale extraction of interpretable, quantitative, morphometric features in non-tumour kidney histology. We use two internal and three external, multi-centre cohorts to analyse over 1000 kidney biopsies and nephrectomies. By associating morphometric features with clinical parameters, we confirm previous concepts and reveal unexpected relations. We show that the extracted features are independent predictors of long-term clinical outcomes in IgA-nephropathy. We introduce single-structure morphometric analysis by applying techniques from single-cell transcriptomics, identifying distinct glomerular populations and morphometric phenotypes along a trajectory of disease progression. Our study provides a concept for Next-generation Morphometry (NGM), enabling comprehensive quantitative pathology data mining, i.e., pathomics.

• MeSH
• glomerulus * patologie   MeSH
• ledviny * patologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: The unphysiological composition of peritoneal dialysis (PD) fluids induces progressive peritoneal fibrosis, hypervascularization and vasculopathy. Information on these alterations after kidney transplantation (KTx) is scant. METHODS: Parietal peritoneal tissues were obtained from 81 pediatric patients with chronic kidney disease stage 5 (CKD5), 72 children on PD with low glucose degradation product (GDP) PD fluids, and from 20 children 4-8 weeks after KTx and preceding low-GDP PD. Tissues were analyzed by digital histomorphometry and quantitative immunohistochemistry. RESULTS: While chronic PD was associated with peritoneal hypervascularization, after KTx vascularization was comparable to CKD5 level. Submesothelial CD45 counts were 40% lower compared with PD, and in multivariable analyses independently associated with microvessel density. In contrast, peritoneal mesothelial denudation, submesothelial thickness and fibrin abundance, number of activated, submesothelial fibroblasts and of mesothelial-mesenchymal transitioned cells were similar after KTx. Diffuse peritoneal podoplanin positivity was present in 40% of the transplanted patients. In subgroups matched for age, PD vintage, dialytic glucose exposure and peritonitis incidence, submesothelial hypoxia-inducible factor 1-alpha abundance and angiopoietin 1/2 ratio were lower after KTx, reflecting vessel maturation, while arteriolar and microvessel p16 and cleaved Casp3 were higher. Submesothelial mast cell count and interleukin-6 were lower, whereas transforming growth factor-beta induced pSMAD2/3 was similar as compared with children on PD. CONCLUSIONS: Peritoneal membrane damage induced with chronic administration of low-GDP PD fluids was less severe after KTx. While peritoneal microvessel density, primarily defining PD transport and ultrafiltration capacity, was normal after KTx and peritoneal inflammation less pronounced, diffuse podoplanin positivity and profibrotic activity were prevalent.

• MeSH
• chronické selhání ledvin * chirurgie   metabolismus   MeSH
• dialýza ledvin MeSH
• dialyzační roztoky metabolismus   MeSH
• dítě MeSH
• glukosa metabolismus   MeSH
• lidé MeSH
• peritoneální dialýza * škodlivé účinky   MeSH
• peritoneum metabolismus   MeSH
• peritonitida * metabolismus   MeSH
• transplantace ledvin * škodlivé účinky   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Funkce pravé komory (PK) má u selhávajícího levého srdce kritický význam. Cílem projektu je studium mechanismů zodpovědných za dysfunkci PK u pacientů s pokročilým srdečním selháním (SS) před transplantací srdce (Tx, n=70), implantací mechanické podpory (n=40) a u kontrol (n=10). Funkce PK bude kvantifikována pomocí metody nezávislé na zátěži (invazivní hemodynamické měření v kombinaci se SPECT tomografickou rekonstrukcí PK) získaných vstupně při kontrolním vyšetření za 6-8 měsíců. Budou studovány klinické, biochemické a hemodynamické faktory spojené se změnou funkce PK během času sledování. Za účelem studia procesů odpovědných za selhání PK, budou získávány vzorky explantovaného myokardu pacientů podstupujících Tx (s či bez dysfunkce PK, n=60) a kontrol (dárci chlopní, n=20). Pomocí kvantitativní histomorfometrie a in-vitro studia mitochondriální respirace chceme určit, jaké jsou vztahy mezi mitochondriální metabolickou aktivitou, ztrátou kapilárního řečiště (rarefakcí) a rozvojem mechanické dysfunkce PK u pacientů se SS.; Function of the right ventricle (RV) is critically important for patients with left heart failure (HF). The aim of the project is to study mechanisms of RV dysfunction in advanced HF prior heart transplantation (Tx, n=70) or LVAD implantation (n=40) and in controls (n=10). RV function will be quantified by comprehensive load-independent method (invasive hemodynamics and SPECT-derived tomographic reconstruction) during altered loading conditions (leg rise, pulmonary vasodilatation) at baseline and at follow-up (6-8 months). Clinical, biochemical and hemodynamic factors associated with RV function change over the time will be analyzed. To better understand processes responsible for progression of RV failure, explanted RV and LV myocardial tissue will be obtained from controls (valve donors, n=20) and HF patients (with or without RV dysfunction, n=60) undergoing Tx. The goal is to address the relations between RV function, loss of myocardial capillary network (rarefaction) and mitochondrial metabolic activity, using quantitative histomorphometry and in vitro mitochondrial assessment.

• MeSH
• biochemie MeSH
• dysfunkce pravé srdeční komory etiologie   MeSH
• hemodynamické monitorování MeSH
• mikrocévy patologie   MeSH
• mikrovaskulární rarefakce MeSH
• mitochondrie MeSH
• nemoci cév MeSH
• plicní hypertenze MeSH
• srdeční selhání komplikace   MeSH
• transplantace srdce MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• kardiologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

BACKGROUND: The rate of reparative osteogenesis controls when an implant is sufficiently stable as to allow functional loading. Using a mini pig model, the rate of reparative osteogenesis in two types of implant sites for example, an osteotomy versus a fresh extraction socket were compared. METHODS: Eight adult mini pigs were used for the study. In phase I, three premolars were extracted on one side of the oral cavity; 12 weeks later, in phase II, osteotomies were produced in healed extraction sites, and contralateral premolars were extracted. Animals were sacrificed 1, 5, and 12 weeks after phase II. Bone repair and remodeling were evaluated using quantitative micro-computed tomographic imaging, histology, and histochemical assays coupled with quantitative dynamic histomorphometry. RESULTS: One week after surgery, extraction sockets and osteotomy sites exhibited similar patterns of new bone deposition. Five weeks after surgery, mineral apposition rates (MARs) were elevated at the injury sites relative to intact bone. Twelve weeks after surgery, the density of new bone in both injury sites was equivalent to intact bone but quantitative dynamic histomorphometry and cellular activity assays demonstrated bone remodeling was still underway. CONCLUSIONS: The mechanisms and rates of reparative osteogenesis were equivalent between fresh extraction sockets and osteotomies. The volume of new bone required to fill a socket, however, was significantly greater than the volume required to fill an osteotomy. These data provide a framework for estimating the rate of reparative osteogenesis and the time to loading of implants placed in healed sites versus fresh extraction sockets.

• MeSH
• endoseální implantace zubů metody   MeSH
• extrakce zubů metody   MeSH
• miniaturní prasata MeSH
• prasata MeSH
• premolár chirurgie   MeSH
• remodelace kosti MeSH
• zubní implantáty * MeSH
• zubní lůžko * diagnostické zobrazování   chirurgie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The effect of peritoneal dialysates with low-glucose degradation products on peritoneal membrane morphology is largely unknown, with functional relevancy predominantly derived from experimental studies. To investigate this, we performed automated quantitative histomorphometry and molecular analyses on 256 standardized peritoneal and 172 omental specimens from 56 children with normal renal function, 90 children with end-stage kidney disease at time of catheter insertion, and 82 children undergoing peritoneal dialysis using dialysates with low-glucose degradation products. Follow-up biopsies were obtained from 24 children after a median peritoneal dialysis of 13 months. Prior to dialysis, mild parietal peritoneal inflammation, epithelial-mesenchymal transition and vasculopathy were present. After up to six and 12 months of peritoneal dialysis, blood microvessel density was 110 and 93% higher, endothelial surface area per peritoneal volume 137 and 95% greater, and submesothelial thickness 23 and 58% greater, respectively. Subsequent peritoneal changes were less pronounced. Mesothelial cell coverage was lower and vasculopathy advanced, whereas lymphatic vessel density was unchanged. Morphological changes were accompanied by early fibroblast activation, leukocyte and macrophage infiltration, diffuse podoplanin presence, epithelial mesenchymal transdifferentiation, and by increased proangiogenic and profibrotic cytokine abundance. These transformative changes were confirmed by intraindividual comparisons. Peritoneal microvascular density correlated with peritoneal small-molecular transport function by uni- and multivariate analysis. Thus, in children on peritoneal dialysis neutral pH dialysates containing low-glucose degradation products induce early peritoneal inflammation, fibroblast activation, epithelial-mesenchymal transition and marked angiogenesis, which determines the PD membrane transport function.

• MeSH
• biopsie MeSH
• chronické selhání ledvin terapie   MeSH
• dialyzační roztoky chemie   toxicita   MeSH
• dítě MeSH
• epitelo-mezenchymální tranzice účinky léků   MeSH
• fibróza MeSH
• glukosa metabolismus   MeSH
• kojenec MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• mladiství MeSH
• peritoneální dialýza škodlivé účinky   MeSH
• peritoneum krevní zásobení   účinky léků   patologie   MeSH
• peritonitida chemicky indukované   patologie   MeSH
• předškolní dítě MeSH
• studie případů a kontrol MeSH
• výsledek terapie MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• práce podpořená grantem MeSH

BACKGROUND: Ultrafiltration decline is a progressive issue for patients on chronic peritoneal dialysis (PD) and can be caused by peritoneal angiogenesis induced by PD fluids. A recent pediatric trial suggests better preservation of ultrafiltration with bicarbonate versus lactate buffered fluid; underlying molecular mechanisms are unknown. METHODS: Angiogenic cytokine profile, tube formation capacity and Receptor Tyrosine Kinase translocation were assessed in primary human umbilical vein endothelial cells following incubation with bicarbonate (BPDF) and lactate buffered (LPDF), pH neutral PD fluid with low glucose degradation product content and lactate buffered, acidic PD fluid with high glucose degradation product content (CPDF). Peritoneal biopsies from age-, PD-vintage- and dialytic glucose exposure matched, peritonitis-free children on chronic PD underwent automated histomorphometry and immunohistochemistry. RESULTS: In endothelial cells angiopoietin-1 mRNA and protein abundance increased 200% upon incubation with BPDF, but decreased by 70% with LPDF as compared to medium control; angiopoietin-2 remained unchanged. Angiopoietin-1/Angiopoietin-2 protein ratio was 15 and 3-fold increased with BPDF compared to LPDF and medium. Time-lapse microscopy with automated network analysis demonstrated less endothelial cell tube formation with BPDF compared to LPDF and CPDF incubation. Receptor Tyrosine Kinase translocated to the cell membrane in BPDF but not in LPDF or CPDF incubated endothelial cells. In children dialyzed with BPDF peritoneal vessels were larger and angiopoietin-1 abundance in CD31 positive endothelium higher compared to children treated with LPDF. CONCLUSION: Bicarbonate buffered PD fluid promotes vessel maturation via upregulation of angiopoietin-1 in vitro and in children on dialysis. Our findings suggest a molecular mechanism for the observed superior preservation of ultrafiltration capacity with bicarbonate buffered PD fluid with low glucose degradation product content.

• MeSH
• angiopoetin-1 metabolismus   MeSH
• angiopoetin-2 metabolismus   MeSH
• antigeny CD31 metabolismus   MeSH
• biopsie MeSH
• chronická nemoc MeSH
• cytokiny metabolismus   MeSH
• dítě MeSH
• endoteliální buňky pupečníkové žíly (lidské) MeSH
• endoteliální buňky metabolismus   MeSH
• glukosa chemie   MeSH
• hydrogenuhličitany chemie   MeSH
• koncentrace vodíkových iontů MeSH
• laktáty chemie   MeSH
• lidé MeSH
• mladiství MeSH
• nemoci ledvin terapie   MeSH
• peritoneální dialýza * MeSH
• peritoneum patologie   MeSH
• pufry * MeSH
• tyrosinkinasové receptory metabolismus   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Isoquercitrin (quercetin-3-O-β-d-glucopyranoside) is a flavonoid that exhibited antioxidant and anti-inflammatory activities in a number of in vitro and in vivo studies. Experimental evidence from rodent models of inflammatory bowel disease is, however, lacking. This study was designed to examine whether isoquercitrin effectively and dose-dependently attenuates acute dextran sulfate sodium (DSS)-induced rat colitis. METHODS: Wistar rats were divided into negative control group (exposed to vehicle only), positive control group (DSS-induced colitis plus vehicle), low isoquercitrin group (DSS pretreated with isoquercitrin 1mg/kg/day) and high isoquercitrin group (DSS with isoquercitrin 10mg/kg/day). Isoquercitrin was administered daily for 14days, and during the last 7days rats drank DSS solution. The effect of isoquercitrin on DSS-induced colitis was assessed clinically (e.g. disease activity index), biochemically (tissue myeloperoxidase activity, local cyclooxygenase-2 expression), using histology (standard hematoxylin-eosin-based histomorphometry, immunohistochemical detection of inducible nitric oxide synthase) and hematology (blood count). RESULTS: Isoquercitrin dose-dependently ameliorated whole colon shortening and mitigated DSS-induced expression of cyclooxygenase-2 and inducible nitric oxide synthase in the descending segment of the organ. However, when different parts of colon were assessed histomorphometrically, the results did not globally support the protective role of this flavonoid. Tissue healing trends observable in the descending colon were not apparent in the rectum, where histological damage was most severe. CONCLUSIONS: We surmise that isoquercitrin may be effective in the prevention of acute colitis. Besides being dose-dependent, the potency of orally administered isoquercitrin may depend on the severity of tissue damage and/or on the site of its action.

• MeSH
• kolitida chemicky indukované   patologie   prevence a kontrola   MeSH
• krysa rodu rattus MeSH
• náhodné rozdělení MeSH
• ochranné látky terapeutické užití   MeSH
• potkani Wistar MeSH
• quercetin analogy a deriváty   terapeutické užití   MeSH
• síran dextranu toxicita   MeSH
• stupeň závažnosti nemoci * MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Náplní projektu je zlepšení diagnostiky a monitorování MKN s upřesněním indikací antiresorpční léčby. K posouzení vhodných cytokinů a chemokinů (sclerostin, NF-kB, TRAP-5a, SDF-1, Runx2/Cbfa1, sFRP, Activin A, Annexin 2, p62, MMP-9) bude využito dostupného biologického materiálu (krevního séra a plazmy kostní dřeně). Bude provedena korelace parametrů kostního metabolismu se zobrazovacími technikami a histomorfometrií vzorku kostní dřeně. Získané výsledky budou sloužit k identifikaci jedinců s MGNV s vysokým rizikem přechodu do symptomatické formy myelomu a k odhalení skupiny nemocných, indikovaných k časnému nasazení antiresorpční terapie ještě před vznikem závažného kostního postižení podchyceného dostupnými moderními zobrazovacími technikami. U jedinců s MM budou výsledky sloužit k vyčlenění skupin nemocných vyžadujících specifickou antiresorpční terapii (bisfosfonáty) a skupiny, u níž nemusí být její dlouhodobé podávání přínosné. Cílem je personalizovaný léčebný přístup se zlepšením kvality života.; The aim of the project is to improve diagnostics and monitoring of myeloma bone disease with specification of the indications of anti-resorption therapy. For the assessment of convenient cytokines and chemokines we will use accessible biological material (blood serum and bone marrow plasma). In selected patients we will perform bone biopsy with the correlation of bone metabolism parameters with bone histomorphometry. The results will identify individuals with MGUS who are at high risk of transformation into asymptomatic and symptomatic MM, and to unravel a group of patients who should be indicated for early administration of anti-resorption therapy (bisphosphonates) before the development of bone involvement assessable by temporary imaging methods. In patients with MM the results will help to differentiate groups of individuals with different severity of skeletal involvement, depicting also patients who do and who possibly do not profit from prolonged bisphosphonates administration in MM remission.

• MeSH
• chemokiny MeSH
• cytokiny MeSH
• individualizovaná medicína MeSH
• inhibitory kostní resorpce MeSH
• kvalita života MeSH
• mnohočetný myelom diagnóza   MeSH
• monoklonální gamapatie nejasného významu MeSH
• nádory kostní dřeně MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• hematologie a transfuzní lékařství
• onkologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR