Fluorescence-based microscopy as one of the standard tools in biomedical research benefits more and more from super-resolution methods, which offer enhanced spatial resolution allowing insights into new biological processes. A typical drawback of using these methods is the need for new, complex optical set-ups. This becomes even more significant when using two-photon fluorescence excitation, which offers deep tissue imaging and excellent z-sectioning. We show that the generation of striped-illumination patterns in two-photon laser scanning microscopy can readily be exploited for achieving optical super-resolution and contrast enhancement using open-source image reconstruction software. The special appeal of this approach is that even in the case of a commercial two-photon laser scanning microscope no optomechanical modifications are required to achieve this modality. Modifying the scanning software with a custom-written macro to address the scanning mirrors in combination with rapid intensity switching by an electro-optic modulator is sufficient to accomplish the acquisition of two-photon striped-illumination patterns on an sCMOS camera. We demonstrate and analyse the resulting resolution improvement by applying different recently published image resolution evaluation procedures to the reconstructed filtered widefield and super-resolved images. This article is part of the Theo Murphy meeting issue 'Super-resolution structured illumination microscopy (part 1)'.

• Klíčová slova
• SIM, laser scanning fluorescence microscopy, multi-photon fluorescence excitation, structured illumination microscopy, super-resolution optical microscopy,
• MeSH
• algoritmy MeSH
• Convallaria ultrastruktura   MeSH
• ledviny ultrastruktura   MeSH
• mikroskopie fluorescenční multifotonová přístrojové vybavení   metody   statistika a číselné údaje   MeSH
• myši MeSH
• optické jevy MeSH
• optické prostředky MeSH
• počítačové zpracování obrazu metody   statistika a číselné údaje   MeSH
• software MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Lead is well known environmental pollutant, which can cause toxic effects in multiple organ systems. However, the influence of lead oxide nanoparticles, frequently emitted to the environment by high temperature technological processes, is still concealed. Therefore, we investigate lead oxide nanoparticle distribution through the body upon their entry into lungs and determine the microscopic and ultramicroscopic changes caused by the nanoparticles in primary and secondary target organs. METHODS: Adult female mice (ICR strain) were continuously exposed to lead oxide nanoparticles (PbO-NPs) with an average concentration approximately 106 particles/cm3 for 6 weeks (24 h/day, 7 days/week). At the end of the exposure period, lung, brain, liver, kidney, spleen, and blood were collected for chemical, histological, immunohistochemical and electron microscopic analyses. RESULTS: Lead content was found to be the highest in the kidney and lungs, followed by the liver and spleen; the smallest content of lead was found in brain. Nanoparticles were located in all analysed tissues and their highest number was found in the lung and liver. Kidney, spleen and brain contained lower number of nanoparticles, being about the same in all three organs. Lungs of animals exposed to lead oxide nanoparticles exhibited hyperaemia, small areas of atelectasis, alveolar emphysema, focal acute catarrhal bronchiolitis and also haemostasis with presence of siderophages in some animals. Nanoparticles were located in phagosomes or formed clusters within cytoplasmic vesicles. In the liver, lead oxide nanoparticle exposure caused hepatic remodeling with enlargement and hydropic degeneration of hepatocytes, centrilobular hypertrophy of hepatocytes with karyomegaly, areas of hepatic necrosis, occasional periportal inflammation, and extensive accumulation of lipid droplets. Nanoparticles were accumulated within mitochondria and peroxisomes forming aggregates enveloped by an electron-dense mitochondrial matrix. Only in some kidney samples, we observed areas of inflammatory infiltrates around renal corpuscles, tubules or vessels in the cortex. Lead oxide nanoparticles were dispersed in the cytoplasm, but not within cell organelles. There were no significant morphological changes in the spleen as a secondary target organ. Thus, pathological changes correlated with the amount of nanoparticles found in cells rather than with the concentration of lead in a given organ. CONCLUSIONS: Sub-chronic exposure to lead oxide nanoparticles has profound negative effects at both cellular and tissue levels. Notably, the fate and arrangement of lead oxide nanoparticles were dependent on the type of organs.

• Klíčová slova
• Brain, Electron microscopy, Inhalation, Kidney, Lead oxide, Liver, Lung, Nanoparticles, Spleen, Toxicity,
• MeSH
• hodnocení rizik MeSH
• inhalační expozice MeSH
• játra účinky léků   metabolismus   ultrastruktura   MeSH
• kovové nanočástice * aplikace a dávkování   chemie   toxicita   MeSH
• látky znečišťující životní prostředí aplikace a dávkování   chemie   farmakokinetika   toxicita   MeSH
• ledviny účinky léků   metabolismus   ultrastruktura   MeSH
• mozek účinky léků   metabolismus   ultrastruktura   MeSH
• myši inbrední ICR MeSH
• olovo aplikace a dávkování   chemie   farmakokinetika   toxicita   MeSH
• oxidy aplikace a dávkování   chemie   farmakokinetika   toxicita   MeSH
• plíce účinky léků   metabolismus   ultrastruktura   MeSH
• slezina účinky léků   metabolismus   ultrastruktura   MeSH
• tkáňová distribuce MeSH
• toxikokinetika MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• látky znečišťující životní prostředí MeSH
• lead oxide MeSH Prohlížeč
• olovo MeSH
• oxidy MeSH

The increasing amount of heavy metals used in manufacturing equivalently increases hazards of environmental pollution by industrial products such as cadmium oxide (CdO) nanoparticles. Here, we aimed to unravel the CdO nanoparticle destiny upon their entry into lungs by inhalations, with the main focus on the ultrastructural changes that the nanoparticles may cause to tissues of the primary and secondary target organs. We indeed found the CdO nanoparticles to be transported from the lungs into secondary target organs by blood. In lungs, inhaled CdO nanoparticles caused significant alterations in parenchyma tissue including hyperemia, enlarged pulmonary septa, congested capillaries, alveolar emphysema and small areas of atelectasis. Nanoparticles were observed in the cytoplasm of cells lining bronchioles, in the alveolar spaces as well as inside the membranous pneumocytes and in phagosomes of lung macrophages. Nanoparticles even penetrated through the membrane into some organelles including mitochondria and they also accumulated in the cytoplasmic vesicles. In livers, inhalation caused periportal inflammation and local hepatic necrosis. Only minor changes such as diffusely thickened filtration membrane with intramembranous electron dense deposits were observed in kidney. Taken together, inhaled CdO nanoparticles not only accumulated in lungs but they were also transported to other organs causing serious damage at tissue as well as cellular level.

• Klíčová slova
• cadmium oxide, electron microscopy, inhalation, kidney, liver, lung, nanoparticles, spleen, toxicity,
• MeSH
• játra metabolismus   patologie   ultrastruktura   MeSH
• kadmium škodlivé účinky   krev   MeSH
• ledviny metabolismus   patologie   ultrastruktura   MeSH
• myši MeSH
• nadechnutí * MeSH
• nanočástice škodlivé účinky   chemie   MeSH
• oxidy škodlivé účinky   krev   chemie   metabolismus   MeSH
• plíce metabolismus   patologie   ultrastruktura   MeSH
• slezina metabolismus   patologie   ultrastruktura   MeSH
• sloučeniny kadmia škodlivé účinky   krev   chemie   metabolismus   MeSH
• velikost částic MeSH
• vystavení vlivu životního prostředí MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cadmium oxide MeSH Prohlížeč
• kadmium MeSH
• oxidy MeSH
• sloučeniny kadmia MeSH

TRIAL DESIGN: This analysis characterizes the degree of early organ involvement in a cohort of oligo-symptomatic untreated young patients with Fabry disease enrolled in an ongoing randomized, open-label, parallel-group, phase 3B clinical trial. METHODS: Males aged 5-18 years with complete α-galactosidase A deficiency, without symptoms of major organ damage, were enrolled in a phase 3B trial evaluating two doses of agalsidase beta. Baseline disease characteristics of 31 eligible patients (median age 12 years) were studied, including cellular globotriaosylceramide (GL-3) accumulation in skin (n = 31) and kidney biopsy (n = 6; median age 15 years; range 13-17 years), renal function, and glycolipid levels (plasma, urine). RESULTS: Plasma and urinary GL-3 levels were abnormal in 25 of 30 and 31 of 31 patients, respectively. Plasma lyso-GL-3 was elevated in all patients. GL-3 accumulation was documented in superficial skin capillary endothelial cells (23/31 patients) and deep vessel endothelial cells (23/29 patients). The mean glomerular filtration rate (GFR), measured by plasma disappearance of iohexol, was 118.1 mL/min/1.73 m(2) (range 90.4-161.0 mL/min/1.73 m(2)) and the median urinary albumin/creatinine ratio was 10 mg/g (range 4.0-27.0 mg/g). On electron microscopy, renal biopsy revealed GL-3 accumulation in all glomerular cell types (podocytes and parietal, endothelial, and mesangial cells), as well as in peritubular capillary and non-capillary endothelial, interstitial, vascular smooth muscle, and distal tubules/collecting duct cells. Lesions indicative of early Fabry arteriopathy and segmental effacement of podocyte foot processes were found in all 6 patients. CONCLUSIONS: These data reveal that in this small cohort of children with Fabry disease, histological evidence of GL-3 accumulation, and cellular and vascular injury are present in renal tissues at very early stages of the disease, and are noted before onset of microalbuminuria and development of clinically significant renal events (e.g. reduced GFR). These data give additional support to the consideration of early initiation of enzyme replacement therapy, potentially improving long-term outcome. TRIAL REGISTRATION: ClinicalTrials.gov NCT00701415.

• MeSH
• biopsie MeSH
• cévní endotel patologie   MeSH
• demografie MeSH
• dítě MeSH
• Fabryho nemoc krev   farmakoterapie   patofyziologie   moč   MeSH
• genotyp MeSH
• glykolipidy krev   MeSH
• hodnoty glomerulární filtrace MeSH
• johexol MeSH
• kůže krevní zásobení   MeSH
• kvalita života MeSH
• ledviny patologie   patofyziologie   ultrastruktura   MeSH
• lidé MeSH
• mladiství MeSH
• mozek patologie   MeSH
• mutace genetika   MeSH
• předškolní dítě MeSH
• sfingolipidy krev   MeSH
• trihexosylceramidy krev   genetika   moč   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• randomizované kontrolované studie MeSH
• Názvy látek
• globotriaosyl lysosphingolipid MeSH Prohlížeč
• globotriaosylceramide MeSH Prohlížeč
• glykolipidy MeSH
• johexol MeSH
• sfingolipidy MeSH
• trihexosylceramidy MeSH

Through linkage analysis and candidate gene sequencing, we identified three unrelated families with the autosomal-dominant inheritance of early onset anemia, hypouricosuric hyperuricemia, progressive kidney failure, and mutations resulting either in the deletion (p.Leu16del) or the amino acid exchange (p.Leu16Arg) of a single leucine residue in the signal sequence of renin. Both mutations decrease signal sequence hydrophobicity and are predicted by bioinformatic analyses to damage targeting and cotranslational translocation of preprorenin into the endoplasmic reticulum (ER). Transfection and in vitro studies confirmed that both mutations affect ER translocation and processing of nascent preprorenin, resulting either in reduced (p.Leu16del) or abolished (p.Leu16Arg) prorenin and renin biosynthesis and secretion. Expression of renin and other components of the renin-angiotensin system was decreased accordingly in kidney biopsy specimens from affected individuals. Cells stably expressing the p.Leu16del protein showed activated ER stress, unfolded protein response, and reduced growth rate. It is likely that expression of the mutant proteins has a dominant toxic effect gradually reducing the viability of renin-expressing cells. This alters the intrarenal renin-angiotensin system and the juxtaglomerular apparatus functionality and leads to nephron dropout and progressive kidney failure. Our findings provide insight into the functionality of renin-angiotensin system and stress the importance of renin analysis in families and individuals with early onset hyperuricemia, anemia, and progressive kidney failure.

• MeSH
• anemie genetika   metabolismus   MeSH
• buněčné linie MeSH
• chronické selhání ledvin genetika   metabolismus   MeSH
• dítě MeSH
• dominantní geny * MeSH
• dospělí MeSH
• genetická vazba MeSH
• hyperurikemie genetika   metabolismus   MeSH
• ledviny cytologie   ultrastruktura   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mutace MeSH
• počítačová simulace MeSH
• předškolní dítě MeSH
• renin genetika   metabolismus   MeSH
• rodokmen MeSH
• sekvenční analýza DNA MeSH
• věk při počátku nemoci MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• renin MeSH

The aim of this study was to determine the copy number changes of chromosomes 7, 17, 3p, and Y in a non-neoplastic tubular epithelium in end-stage kidney disease (ESKD). Seventeen kidneys from 11 patients with ESKD were retrieved from the archive files. Non-neoplastic kidney tissue in these cases was examined separately. Tissues containing papillary adenomas (PA), clear (CRCC) and papillary renal cell carcinomas (PRCC), and myxoid liposarcoma (LPS) were examined using the same probes and compared with non-neoplastic tissue. Tubular changes in the kidney parenchyma were classified into three types: (1) The vast majority of tubules were entirely atrophic; (2) Several tubules were hyperplastic, i.e., tubules with undifferentiated large epithelial cells, in which it was impossible to establish the specific type of a renal tubulus; (3) Dysplastic tubules were dilated, sometimes wrinkled. The basal membranes were lined by large eosinophilic epithelial cells with polymorphic nuclei and pseudostratification. Nucleoli were clearly visible. These tubular changes were multifocal with a haphazard distribution within the atrophic parenchyma. PA were detected in nine patients, of whom eight patients also revealed an additional tumor type(s) (4x CRCC, 3x PRCC, 1x PRCC, and CRCC). One patient had a CRCC only, another had a combination of PRCC and LPS. Chromosomal abnormalities were found in the second and third group of tubular changes, i.e., in hyperplastic and dysplastic tubules. Trisomy of chromosome 7 was detected in six cases, whereas trisomy of chromosome 17 in eight cases. A combination of both trisomies was found in five cases. Loss of chromosome Y was found in two cases. Fluorescence in situ hybridization on tissues containing papillary adenomas, renal cell carcinomas, and liposarcoma revealed expected results, i.e., trisomy of chromosomes 7 and 17 in all PAs and PRCC. No gains were present in CRCC and LPS. Loss of Y was found in six PA, five PRCC, and one LPS; loss of X was found in two CRCCs. We suggest that chromosomal changes typical of the papillary renal cell lesions, i.e., trisomies of chromosomes 7 and 17, are very frequent in non-neoplastic parenchyma of the end-stage kidney, and they have a tendency to a multifocal occurrence.

• MeSH
• adenom genetika   MeSH
• chronické selhání ledvin genetika   patologie   MeSH
• dospělí MeSH
• hybridizace in situ fluorescenční MeSH
• karcinom z renálních buněk genetika   MeSH
• ledvinové kanálky patologie   MeSH
• ledviny ultrastruktura   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lidské chromozomy, pár 17 genetika   MeSH
• lidské chromozomy, pár 7 genetika   MeSH
• lidský chromozom Y genetika   MeSH
• myxoidní liposarkom genetika   MeSH
• senioři MeSH
• trizomie genetika   patologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Autosomal dominant hyperuricemia, gout, renal cysts, and progressive renal insufficiency are hallmarks of a disease complex comprising familial juvenile hyperuricemic nephropathy and medullary cystic kidney diseases type 1 and type 2. In some families the disease is associated with mutations of the gene coding for uromodulin, but the link between the genetic heterogeneity and mechanism(s) leading to the common phenotype symptoms is not clear. In 19 families, we investigated relevant biochemical parameters, performed linkage analysis to known disease loci, sequenced uromodulin gene, expressed and characterized mutant uromodulin proteins, and performed immunohistochemical and electronoptical investigation in kidney tissues. We proved genetic heterogeneity of the disease. Uromodulin mutations were identified in six families. Expressed, mutant proteins showed distinct glycosylation patterns, impaired intracellular trafficking, and decreased ability to be exposed on the plasma membrane, which corresponded with the observations in the patient's kidney tissue. We found a reduction in urinary uromodulin excretion as a common feature shared by almost all of the families. This was associated with case-specific differences in the uromodulin immunohistochemical staining patterns in kidney. Our results suggest that various genetic defects interfere with uromodulin biology, which could lead to the development of the common disease phenotype. 'Uromodulin-associated kidney diseases' may be thus a more appropriate term for this syndrome.

• MeSH
• bazální membrána patologie   ultrastruktura   MeSH
• biopsie MeSH
• dítě MeSH
• dna (nemoc) MeSH
• dospělí MeSH
• genetická heterogenita * MeSH
• genetická vazba MeSH
• hyperurikemie genetika   metabolismus   MeSH
• hypofýza cytologie   MeSH
• imunohistochemie MeSH
• kultivované buňky MeSH
• ledvinové kanálky patologie   ultrastruktura   MeSH
• ledviny metabolismus   patologie   chirurgie   ultrastruktura   MeSH
• lidé MeSH
• lidské chromozomy, pár 1 MeSH
• lidské chromozomy, pár 16 MeSH
• missense mutace MeSH
• mladiství MeSH
• mukoproteiny genetika   metabolismus   moč   MeSH
• mutační analýza DNA MeSH
• polycystické ledviny autozomálně dominantní genetika   MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• rodokmen MeSH
• sekvence nukleotidů MeSH
• syndrom MeSH
• transfekce MeSH
• uromodulin MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• mukoproteiny MeSH
• UMOD protein, human MeSH Prohlížeč
• uromodulin MeSH

One of four Hoplobatrachus occipitalis (Günther, 1859) frogs received from Niger, West Africa was heavily infected with Lankesterella blood and pre-erythrocytic stages. Infected blood and tissues from this frog were force-fed to the remaining three frogs. Two survived to necropsy on days 14 and 27 post-feeding and were found to be infected with gamogonic and oogonic stages, respectively. The source of infection is inconclusive, as a natural origin cannot be excluded. Microgamont, macrogamont, oocyst and sporozoite structure and fine structure are described and found to conform in general, but not in detail, to previous descriptions. Gamonts and oocysts occurred predominantly in the liver and spleen. Walled sporulating oocysts were situated within macrophage centres. Oocysts yielded a progeny of 32 sporozoites. Pre-erythrocytic sporozoites developed within expanded inclusions, within their host cell, from which they massively invaded the liver and spleen, and to a lesser extent the lungs and kidneys. Sporozoites occurred in a parasitophorous vacuole in the erythrocytes. Conspecificity with Lankesterella dicroglossi Paperna et Ogara, 1996 reported from the same host species in Kenya remains uncertain due to several structural and developmental differences.

• MeSH
• Apicomplexa růst a vývoj   patogenita   ultrastruktura   MeSH
• druhová specificita MeSH
• erytrocyty parazitologie   ultrastruktura   MeSH
• infekce přenášené vektorem MeSH
• interakce hostitele a parazita MeSH
• játra parazitologie   ultrastruktura   MeSH
• ledviny parazitologie   ultrastruktura   MeSH
• makrofágy parazitologie   ultrastruktura   MeSH
• plíce parazitologie   ultrastruktura   MeSH
• protozoální infekce přenos   MeSH
• Ranidae parazitologie   MeSH
• rastrovací tunelová mikroskopie MeSH
• slezina parazitologie   ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Niger MeSH

An overview of immunohistological and molecular genetic methods for diagnosis of Alport syndrome (AS) is given with practical experience from groups of authors' observations. Immunofluorescent investigation using antibodies against alfa chains of collagen IV was performed on cryostat sections from 29 punction nephrobiopsies and 9 skin excisions taken for support of differential diagnosis of AS particularly against the thin membranes glomerulopathy. Alfa chains deviations in other renal diseases were followed in another 14 cases. Molecular genetical investigation of AS by an indirect DNA diagnostics was performed in 35 families with presumed AS and in 27 patients with probable mutation a mutation screening of COL4AS gene by a direct method SSCP was made. The mutation was proved in 10 cases. Because of genotypical and phenotypical variability of AS the diagnostic gain only increases when all the accessible methods are combined.

• MeSH
• dědičná nefritida diagnóza   genetika   patologie   MeSH
• diferenciální diagnóza MeSH
• dítě MeSH
• genetické markery MeSH
• imunohistochemie MeSH
• kolagen analýza   MeSH
• ledviny ultrastruktura   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• polymorfismus konformace jednovláknové DNA MeSH
• předškolní dítě MeSH
• Check Tag
• dítě MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• genetické markery MeSH
• kolagen MeSH

The objective of the paper is to draw attention to a rare cause of rapidly progressing renal failure which developed in the course of four months as a result of light chain deposition disease. The authors submit two case-histories of the disease assessed by renal biopsy after previous clinical and laboratory suspicion of monoclonal gammapathy. In one patient in the sternal punctate plasmacytoma was diagnosed and in the second case it was not possible to detect any type of monoclonal gammapathy or another possible cause of disease. Renal failure was in both cases irreversible and both patients were enlisted in regular haemodialyzation treatment.

• MeSH
• ledviny imunologie   ultrastruktura   MeSH
• lehké řetězce imunoglobulinů metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nemoci ledvin imunologie   patologie   MeSH
• renální insuficience imunologie   patologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• kazuistiky MeSH
• Názvy látek
• lehké řetězce imunoglobulinů MeSH