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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
Uterine sarcomas with KAT6B/A::KANSL1 fusion represent a new entity characterized by bland morphology, commonly with hybrid features of low-grade endometrial stromal sarcoma (LG-ESS) and tumors with smooth muscle differentiation. In our study, we performed a detailed morphological, immunohistochemical, and molecular analysis of 9 cases of these tumors. Six of those had been originally diagnosed as LG-ESS, one as leiomyoma, one as leiomyosarcoma, and the remaining case as sarcoma with the KAT6B/A::KANSL1 fusion. Seven cases showed overlapping features between endometrial stromal and smooth muscle tumors, one case resembled cellular leiomyoma, and one case resembled high-grade endometrial stromal sarcoma. Immunohistochemically, the tumors showed a common expression of smooth muscle markers and endometrial stromal markers. Molecular findings showed the KAT6B/A::KANSL1 fusion in all cases (by NGS and FISH). In addition, mutations affecting genes such as TP53, PDGFRB, NF1, RB1, PTEN, ATM, RB1, FANCD2, and TSC1 were present in all 5 cases with aggressive behavior. One patient with no evidence of disease showed no additional mutations, while another harbored a mutation of a single gene (ERCC3). Of the 8 patients with available follow-up, two died of disease, 3 are currently alive with disease, and 3 have no evidence of disease. The correct recognition of tumors with the KAT6B/A::KANSL1 fusion is essential because despite the bland morphological features of most cases, these tumors have a propensity for aggressive behavior.
- MeSH
- dospělí MeSH
- endometriální stromální sarkom genetika patologie MeSH
- fúzní onkogenní proteiny genetika MeSH
- histonacetyltransferasy genetika MeSH
- imunohistochemie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mutace MeSH
- nádorové biomarkery * genetika analýza MeSH
- nádory dělohy * patologie genetika MeSH
- sarkom genetika patologie MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
TFE3 rearrangements characterize histogenetically, topographically, and biologically diverse neoplasms. Besides being a universal defining feature in alveolar soft part sarcoma (ASPS) and clear cell stromal tumor of the lung, TFE3 fusions have been reported in subsets of renal cell carcinoma, perivascular epithelioid cell tumor (PEComa), epithelioid hemangioendothelioma and ossifying fibromyxoid tumors. TFE3 -related neoplasms are rare in the head and neck and may pose diagnostic challenges. We herein describe 22 TFE3 fusion neoplasms affecting 11 males and 11 females aged 4 to 79 years (median, 25) and involving different head and neck sites: sinonasal cavities (n = 8), tongue (n = 4), oral cavity/oropharynx (n = 3), salivary glands (n = 2), orbit (n = 2), and soft tissue or unspecified sites (n = 3). Based on morphology and myomelanocytic immunophenotype, 10 tumors qualified as ASPS, 7 as PEComas (3 melanotic; all sinonasal), and 5 showed intermediate (indeterminate) histology overlapping with ASPS and PEComa. Immunohistochemistry for TFE3 was homogeneously strongly positive in all cases. Targeted RNA sequencing/FISH testing confirmed TFE3 fusions in 14 of 16 successfully tested cases (88%). ASPSCR1 was the most frequent fusion partner in ASPS (4 of 5 cases); one ASPS had a rare VCP::TFE3 fusion. The 6 successfully tested PEComas had known fusion partners as reported in renal cell carcinoma and PEComas ( NONO, PRCC, SFPQ , and PSPC1 ). The indeterminate tumors harbored ASPSCR1::TFE3 (n = 2) and U2AF2::TFE3 (n = 1) fusions, respectively. This large series devoted to TFE3-positive head and neck tumors illustrates the recently proposed morphologic overlap in the spectrum of TFE3 -associated mesenchymal neoplasms. While all PEComas were sinonasal, ASPS was never sinonasal and occurred in diverse head and neck sites with a predilection for the tongue. The indeterminate (PEComa-like) category is molecularly more akin to ASPS but shows different age, sex, and anatomic distribution compared with classic ASPS. We report VCP as a novel fusion partner in ASPS and PSPC1 as a novel TFE3 fusion partner in PEComa (detected in one PEComa). Future studies should shed light on the most appropriate terminological subtyping of these highly overlapping tumors.
- MeSH
- alveolární sarkom měkkých tkání * genetika patologie MeSH
- dítě MeSH
- dospělí MeSH
- fenotyp MeSH
- genetická predispozice k nemoci MeSH
- genová přestavba * MeSH
- hybridizace in situ fluorescenční MeSH
- imunohistochemie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- nádorové biomarkery * genetika analýza MeSH
- nádory hlavy a krku * genetika patologie chemie MeSH
- nádory z perivaskulárních epiteloidních buněk * genetika patologie chemie MeSH
- předškolní dítě MeSH
- senioři MeSH
- transkripční faktory BHLH-Zip * genetika MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
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
A novel Gram-stain-negative, strictly aerobic, rod-shaped, light-yellow-pigmented, and chemo-organoheterotrophic bacterium, designated DF-77T, was isolated from dense mats of filamentous algae collected in March 2004 at Okinawa in Japan. The microorganism grew at 0-2.0% NaCl concentrations (w/v), pH 6.0-9.0, and 20-30 °C. The 16S rRNA gene sequence-based phylogenetic tree demonstrated that the strain DF-77T is a novel member of the family Flavobacteriaceae and was greatly related to Flagellimonas nanhaiensis SM1704T with sequence similarity of 95.5%. The main fatty acids were iso-C15:1 G, iso-C15:0, and iso-C17:0 3-OH, and the only isoprenoid quinone was menaquinone-6. The dominant polar lipids were phosphatidylethanolamine, two unidentified aminolipids, an unidentified phosphoaminolipid, and four unidentified lipids. The genome size of strain DF-77T was 3.60 Mbp with a DNA G + C content of 47.5%. The average nucleotide identity (ANI) value between the genomes of strain DF-77T and its closely related species was 69.8-70.7%. The digital DNA - DNA hybridization (dDDH) value of strain DF-77T with the strain of F. nanhaiensis SM1704T was 16.8%. The genome of the strain DF-77T revealed that it encoded several genes involved in bio-macromolecule degradation, indicating a high potential for producing industrially useful enzymes. Consequently, the strain is described as a new species in the genus Flagellimonas, for which the name Flagellimonas algarum sp. nov., is proposed with the type strain DF-77T (= KCTC 72791T = NBRC 114251T).
- MeSH
- DNA bakterií genetika chemie MeSH
- Flavobacteriaceae * klasifikace izolace a purifikace genetika MeSH
- fosfolipidy analýza MeSH
- fylogeneze MeSH
- genom bakteriální MeSH
- hybridizace nukleových kyselin MeSH
- mastné kyseliny analýza MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- techniky typizace bakterií MeSH
- vitamin K 2 analýza analogy a deriváty MeSH
- zastoupení bazí MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Japonsko MeSH
Cíl: 18 F-fluorestradiol je nové radiofarmakum, které lze použít při zobrazování karcinomu prsu, indikace v klinických postupech ještě nemají své pevné místo, proto je cílem studie posoudit klinický význam zobrazení karcinomu prsu estrogen-pozitivních receptorů (ER+) pomocí 18 F-fluorestradiolu (18 F-FES) PET/CT nebo PET/MR z hlediska využití při rozhodnutí o léčbě. Studie se zabývá také volbou využití PET/CT nebo PET/MR ve stagingu a restagingu karcinomu prsu. Metodika: U 40 pacientek s estrogen pozitivním karcinomem prsu bylo provedeno hybridní zobrazení s intravenózní aplikací 18 F-FES, ve 25 případech bylo použito PET/CT, v 15 případech PET/MR. Radiofarmakum bylo injekčně aplikováno v aktivitě 2,5 MBq/kg. U deseti pacientek byla jako restagingová metoda použito PET/ MR, v pěti případech stagingu před operací bylo provedeno PET/MR s cíleným plným diagnostickým MR zobrazením prsu v pronační poloze s následným zobrazením trupu v poloze na zádech. Všechna vyšetření PET/MR byla provedena po aplikaci gadoliniové kontrastní látky, zobrazení zahrnovalo zobrazení mozku v T1 STARVIBE. PET/CT bylo provedeno kontinuální PET akvizicí následně po akvizici CT s intravenózním podáním jodované kontrastní látky, v pěti případech bylo provedeno ve stagingu, ve 20 případech v restagingu. Výsledky: Nejdůležitějším výsledkem byla detekce ER+ metastáz při negativním výsledku 18 F-FDG-PET (12krát) – včetně mozkových a jaterních metastáz, perzistující ER+ metastáz (7krát), staging onemocnění (10krát), ztráta ER (4krát) a negativní nález pro metastázy (2), při pěti vyšetřeních nebyly nalezeny žádné přidané informace. Závěr: 18 F-FES-PET poskytuje klinicky vý- znamné informace pro volbu léčebné strategie, 18 F-FES-PET/MR je výhodné vyšetření při zaměření na zobrazení mozku a jater s možnos- tí prokázat anebo vyloučit metastázy v těchto orgánech.
Aim. 18 F-fluoroestradiol is a novel radiopharmaceutical useful in the imaging of breast carcinoma, the indications in clinical scenarios are under development. The purpose of the study is to assess the clinical impact of the imaging of the breast carcinoma with estrogenpositive receptors (ER+) using 18 F-fluoroestra- diol ( 18 F-FES) PET/CT or PET/MRI according to the treatment decision making. The study is concerned in the different preference of PET/CT and PET/MRI in the staging and restaging. Methods. 40 patients with estrogen positive breast carcinoma underwent the hybrid imaging after intravenous application of 18 F-FES, in 25 cases it was used PET/CT, in 15 cases PET/ MRI. The radiopharmaceutical was injected with the activity of 2,5 MBq/kg. In 10 patient, PET/MRI was used as restaging method, PET/ MRI was performed in the 5 cases of the staging before surgery with targeted full diagnostic MRI imaging of the breast in prone position, followed by the trunk imaging in supine position. All PET/MRI were performed after application of the gadolinium contrast material, the imaging included brain imaging in T1 STARVIBE. PET/CT was performed using the continuous PET acquisition after CT with the intravenous administration of the iodinated contrast material, in 5 cases was performed in staging, in 20 cases in restaging Results. The most important informa- tion was detection of ER+ metastases when 18 F-FDG-PET was negative (12×) – including brain and liver metastases, the persistent ER+ of the metastases (7×), staging of the disease (10×), the loss of the ER (4x) and the negative finding for metastases (2), no added information was found in 5 examinations. Conclusion. 18 F-FES-PET provided the impor- tant clinical information to treatment strategy, 18 F-FES-PET/MRI improves the imaging of metastases in brain and liver.
PURPOSE: Minimally invasive surgery (MIS) in neonates and infants presents technical challenges and is still unfamiliar to many paediatrics surgeons. This study aims to identify currently available simulators for neonatal/infant MIS training, to assess their validity, level of evidence, and related recommendations. METHODS: The review followed PRISMA guidelines and was registered in PROSPERO (CRD420250581050). Electronic search limited to English articles was performed through PubMed/MEDLINE, SCOPUS, Web of Science and Cochrane Database from January 2010 to June 2024. RESULTS: Out of 1084 identified records, 72 studies met the inclusion criteria and were analysed across general, gastrointestinal, thoracic, and urological MIS specialties. Recent efforts have led to the development of 3D-printed, animal-based, and hybrid models several of which showed high fidelity, skill differentiation, and educational value. Despite promising results, no universal MIS training model exists for neonate/infant patients, highlighting the need for structured, proficiency-based curricula. Overall, studies demonstrated moderate levels of evidence and recommendation, supporting integration of cost-effective simulation into paediatrics MIS training CONCLUSION: This systematic review highlights the need for validated, standardized simulation models and proficiency-based curricula to optimize neonate and infant MIS training and guide future research toward improving model fidelity, accessibility, and long-term educational outcomes.
Allodiploid hybrid species, Aspergillus latus, belonging to section Nidulantes, is a hybrid of A. spinulosporus and an unknown species closely related to A. quadrilineatus and A. sublatus. This hybrid has often been misidentified as the species in section Nidulantes, such as A. nidulans, A. spinulosporus, A. sublatus, or other cryptic species. Aspergillus latus has not been reported in Japan as well as Asia so far. In this study, we screened 23 clinical strains identified as A. spinulosporus isolated in Japan from 2012 to 2023 and found seven A. latus strains. To characterize the A. latus strains, we conducted comprehensive phenotyping including morphological observation, whole genome sequences, and phylogenetic analysis based on calmodulin (CaM) gene. In addition, we conducted antifungal susceptibility testing for A. latus strains. As a result, the morphological characters of A. latus were more similar to those of A. spinulosporus compared to A. sublatus. However, the ascospore of A. latus differed from that of A. spinulosporus. Phylogenetic analysis revealed that different CaM alleles from the same isolate clustered separately with A. spinulosporus and A. sublatus, consistent with its hybrid origin. Furthermore, A. latus strains showed reduced susceptibility to caspofungin and amphotericin B compared to A. spinulosporus, while they were susceptible to azoles. Our results suggest that A. latus has been a causative pathogen of aspergillosis in Japan since 2013.
- MeSH
- antifungální látky farmakologie MeSH
- Aspergillus * genetika klasifikace izolace a purifikace účinky léků MeSH
- aspergilóza * mikrobiologie epidemiologie MeSH
- fylogeneze MeSH
- kalmodulin genetika MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- sekvenování celého genomu MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Japonsko MeSH
UNLABELLED: Despite significant advances in knowledge and the development of guidelines, the management of hypoplastic left heart syndrome (HLHS) remains highly variable. A structured questionnaire was circulated across European Association of Paediatric & Congenital Cardiology (AEPC) affiliated centres. The aims were to evaluate standards in pre-operative assessment, types of surgery, follow-up and medical practices in children with HLHS. Thirty-one centres from 20 countries completed the survey. Delivery of babies with HLHS occurred in co-located maternity hospitals in 74% of centres; 29% were planned for spontaneous onset of labour, while 54% decided on a case-by-case basis. The preferred initial palliation was a right ventricle-pulmonary artery conduit in 55% of cases, modified Blalock-Thomas Taussig shunt (mBTTS) in 35%, and hybrid in 15% of cases. Timing for Glenn varied from 3 to 6 months of age and preoperative examination varied greatly: 65% performed cardiac catheterization and only 19% performed cardiac magnetic resonance. Stage III palliation was performed at a highly variable interval (2-6 years of age), nearly always employing an extracardiac conduit. Fenestration was routinely performed in 61% and reserved for borderline cases in 39%. All the centers adopted warfarin for the first 3-12 months after Fontan completion, and continued if a fenestration was present, while in non-fenestrated aspirin was left by most centers (e.g. 68%). However, there was a high disparity in the use of heart failure medications (e.g. in interstage I-II 35% use ACE-inhibitors, and only 26% digoxin). Follow-up practice also varied widely with only 60% employing specific protocols. CONCLUSION: This first multi-centre European survey from 31 centres from 20 different European countries highlighted a high practice variation in HLHS management across all the stages of Single Ventricle (Fontan) palliation. Major variations pertained to pre- and post-surgical investigations, surgical strategy for stage I and III, medical treatment regimens, and follow-up programs. WHAT IS KNOWN: • Hypoplastic left heart syndrome (HLHS) remains one of the most complex and challenging congenital cardiac defects to manage. • Investigating the management of children with HLHS across different European centres can facilitate study of the most effective management strategies. WHAT IS NEW: • Significant variation in HLHS management were reported in relation to pre- and post-surgical examinations, surgical strategy at stage I and III, medical treatment regimens, and follow-up programs. • Greater standardisation of imaging and diagnostic evaluation, medical treatment and follow-up surveillance may improve outcomes for these vulnerable patients and warrants further study.
- MeSH
- dítě MeSH
- kojenec MeSH
- lékařská praxe - způsoby provádění * statistika a číselné údaje MeSH
- lidé MeSH
- novorozenec MeSH
- paliativní péče MeSH
- předškolní dítě MeSH
- průzkumy a dotazníky MeSH
- syndrom hypoplazie levého srdce * chirurgie diagnóza terapie MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé MeSH
- novorozenec MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- Geografické názvy
- Evropa MeSH
BACKGROUND: The education of healthcare professionals, including nurses, represents a critical intersection with health systems science (HSS), which is often considered the third pillar of healthcare education alongside basic and clinical sciences. Despite the amount of research on nursing education during the COVID-19 pandemic, there remains a gap in analysis from an HSS perspective. METHODS: A Comparative Education Approach involving undergraduate nursing programs (UNPs) across Europe, with longitudinal data collection from 2002 to 2023. The aim of the study was threefold: (a) to summarize the overall changes in UNPs during the COVID-19 pandemic; (b) to identify the changes retained in the post-pandemic era; and (c) to identify the impact of the pandemic on nursing education as perceived by nurse educators across nine European countries. RESULTS: This study compares the changes in nursing education in European countries during and after the COVID-19 pandemic using a qualitative approach with data from nine universities. The COVID-19 pandemic had a significant impact on education provision. During the first wave, government restrictions forced a complete shift to online learning for theory classes, clinical training and laboratories. In subsequent waves, a hybrid format was chosen that combined online and face-to-face sessions. A major challenge was the placement of nursing students alongside general university students. This approach neglected their need for practical clinical training, which is crucial for their future careers. To compensate for the lack of clinical hours, various alternative teaching methods were introduced. Students were also offered the opportunity to volunteer in large-scale public health initiatives such as vaccination and testing campaigns, although bedside care for COVID-19 patients remained limited. The pandemic has also left its mark in the post-pandemic period. Some UNPs have retained elements of online education, notably lectures, research seminars, meetings, consultations and even online exams. Interestingly, an initial increase in applicant numbers was observed at six of the nine participating UNPs. CONCLUSIONS: The COVID-19 pandemic has widened the gap between university-educated nurses and the clinical setting, i.e. between theory and practise, underlining the importance of HSS in nursing education. Rebuilding strong partnerships is crucial, but simply returning to the pre-pandemic model is not enough. To ensure uninterrupted education during future crises, proactive planning, including the creation of predefined protocols for collaboration, is essential. The pandemic underscores the need for closer alignment between the two sectors, which would better equip future nurses with the skills they need to thrive in the nursing workforce and ensure they are prepared for the challenges of the 21st century.
- MeSH
- COVID-19 * epidemiologie MeSH
- distanční studium trendy MeSH
- kurikulum MeSH
- lidé MeSH
- longitudinální studie MeSH
- pandemie MeSH
- SARS-CoV-2 MeSH
- studium ošetřovatelství bakalářské * trendy organizace a řízení MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Geografické názvy
- Evropa MeSH