Single-photon optogenetics enables precise, cell-type-specific modulation of neuronal circuits, making it a crucial tool in neuroscience. Its miniaturization in the form of fully implantable wide-field stimulator arrays enables long-term interrogation of cortical circuits and bears promise for brain-machine interfaces for sensory and motor function restoration. However, achieving selective activation of functional cortical representations poses a challenge, as studies show that targeted optogenetic stimulation results in activity spread beyond one functional domain. While recurrent network mechanisms contribute to activity spread, here we demonstrate with detailed simulations of isolated pyramidal neurons from cats of unknown sex that already neuron morphology causes a complex spread of optogenetic activity at the scale of one cortical column. Since the shape of a neuron impacts its optogenetic response, we find that a single stimulator at the cortical surface recruits a complex spatial distribution of neurons that can be inhomogeneous and vary with stimulation intensity and neuronal morphology across layers. We explore strategies to enhance stimulation precision, finding that optimizing stimulator optics may offer more significant improvements than the preferentially somatic expression of the opsin through genetic targeting. Our results indicate that, with the right optical setup, single-photon optogenetics can precisely activate isolated neurons at the scale of functional cortical domains spanning several hundred micrometers.

• MeSH
• kočky MeSH
• modely neurologické MeSH
• mozková kůra fyziologie   cytologie   MeSH
• neurony fyziologie   MeSH
• optogenetika * metody   MeSH
• pyramidové buňky fyziologie   MeSH
• světelná stimulace metody   MeSH
• zvířata MeSH
• Check Tag
• kočky MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The retrograde approach in chronic total occlusion (CTO) interventions often encounters significant challenges, particularly, when aligning the retrograde microcatheter (MC) with the antegrade system is difficult, complicating or even preventing standard externalization. To address these issues, techniques like the "tip-in" have proven to be effective backup strategies. We introduce the "Manual Microcatheter-tip Modification" (MMM) technique as an alternative when the "tip-in" method faces complications. We present a case of a left anterior descending CTO where MMM was successfully employed for the first time, enabling successful revascularization by manually modifying the MC tip to engage the retrograde guidewire. We explore the technical details within the framework of contemporary CTO PCI. This new technique could enhance the management of CTO interventions, offering innovative solutions when traditional externalization methods are problematic.

• MeSH
• balónková koronární angioplastika přístrojové vybavení   MeSH
• chronická nemoc MeSH
• design vybavení * MeSH
• koronární angiografie * MeSH
• koronární angioplastika přístrojové vybavení   MeSH
• koronární okluze * diagnostické zobrazování   terapie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• miniaturizace * MeSH
• senioři MeSH
• srdeční katetrizace přístrojové vybavení   MeSH
• srdeční katétry * MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• biomedicínský výzkum MeSH
• interdisciplinární výzkum MeSH
• melanom experimentální * MeSH
• miniaturní prasata MeSH
• modely nemocí na zvířatech MeSH
• molekulární biologie MeSH
• osobní vzpomínky jako téma MeSH
• spontánní regrese nádoru MeSH
• Publikační typ
• novinové články MeSH

PURPOSE: To assess the safety and feasibility of direct vitrectomy-sparing subretinal injection for gene delivery in a large animal model. METHODS: The experimental Liběchov minipigs were used for subretinal delivery of a plasmid DNA vector (pS/MAR-CMV-copGFP) with cytomegalovirus (CMV) promoter, green fluorescent protein (GFP) reporter (copGFP) and a scaffold/matrix attachment region (S/MAR) sequence. The eyes were randomized to subretinal injection of the vector following pars plana vitrectomy (control group) or a direct injection without prior vitrectomy surgery (experimental group). Intra- and post-operative observations up to 30 days after surgery were compared. RESULTS: Six eyes of three mini-pigs underwent surgery for delivery into the subretinal space. Two eyes in the control group were operated with a classical approach (lens-sparing vitrectomy and posterior hyaloid detachment). The other four eyes in the experimental group were injected directly with a subretinal cannula without vitrectomy surgery. No adverse events, such as endophthalmitis, retinal detachment and intraocular pressure elevation were observed post-operatively. The eyes in the experimental group had both shorter surgical time and recovery while achieving the same surgical goal. CONCLUSIONS: This pilot study demonstrates that successful subretinal delivery of gene therapy vectors is achievable using a direct injection without prior vitrectomy surgery.

• MeSH
• genetická terapie * metody   MeSH
• genetické vektory * aplikace a dávkování   MeSH
• injekce nitrooční MeSH
• miniaturní prasata * MeSH
• modely nemocí na zvířatech MeSH
• pilotní projekty MeSH
• plazmidy aplikace a dávkování   MeSH
• prasata MeSH
• retina MeSH
• studie proveditelnosti * MeSH
• technika přenosu genů * MeSH
• vitrektomie * metody   MeSH
• zelené fluorescenční proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Anandamide (AEA) is an important modulator of nociception in the spinal dorsal horn, acting presynaptically through Cannabinoid (CB1) and Transient receptor potential vanilloid (TRPV1) receptors. The role of AEA (1 μM, 10 μM, and 30 μM) application on the modulation of nociceptive synaptic transmission under control and inflammatory conditions was studied by recording miniature excitatory postsynaptic currents (mEPSCs) from neurons in spinal cord slices. Inhibition of the CB1 receptors by PF514273, TRPV1 by SB366791, and the fatty acid amide hydrolase (FAAH) by URB597 was used. Under naïve conditions, the AEA application did not affect the mEPSCs frequency (1.43±0.12 Hz) when all the recorded neurons were considered. The mEPSC frequency increased (180.0±39.2%) only when AEA (30 μM) was applied with PF514273 and URB597. Analysis showed that one sub-population of neurons had synaptic input inhibited (39.1% of neurons), the second excited (43.5%), whereas 8.7% showed a mixed effect and 8.7% did not respond to the AEA. With inflammation, the AEA effect was highly inhibitory (72.7%), while the excitation was negligible (9.1%), and 18.2% were not modulated. After inflammation, more neurons (45.0%) responded even to low AEA by mEPSC frequency increase with PF514273/URB597 present. AEA-induced dual (excitatory/inhibitory) effects at the 1st nociceptive synapse should be considered when developing analgesics targeting the endocannabinoid system. These findings contrast the clear inhibitory effects of the AEA precursor 20:4-NAPE application described previously and suggest that modulation of endogenous AEA production may be more favorable for analgesic treatments.

• MeSH
• amidohydrolasy MeSH
• analgetika farmakologie   MeSH
• benzamidy * MeSH
• endokanabinoidy * farmakologie   MeSH
• karbamáty * MeSH
• kyseliny arachidonové * MeSH
• lidé MeSH
• nocicepce * MeSH
• polynenasycené alkamidy farmakologie   MeSH
• zadní rohy míšní MeSH
• zánět farmakoterapie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

We applied Raman spectroscopy to brain and skin tissues from a minipig model of Huntington's disease. Differences were observed between measured spectra of tissues with and without Huntington's disease, for both brain tissue and skin tissue. There are linked to changes in the chemical composition between tissue types. Using machine learning we correctly classified 96% of test spectra as diseased or wild type, indicating that the test would have a similar accuracy when used as a diagnostic tool for the disease. This suggests the technique has great potential in the rapid and accurate diagnosis of Huntington's and other neurodegenerative diseases in a clinical setting.

• MeSH
• Huntingtonova nemoc * diagnóza   MeSH
• lidé MeSH
• miniaturní prasata MeSH
• mozek MeSH
• neurodegenerativní nemoci * MeSH
• prasata MeSH
• Ramanova spektroskopie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The availability of dialysis membranes in the form of hollow fibres with diameters compatible with the fused silica capillaries used in capillary electrophoresis is very limited. However, haemodialysis bicarbonate cartridges commonly used in human medicine containing polysulfone hollow fibres are available on the market and are used for the fabrication of coaxial microdialysis probes. The miniature probe design ensures that steady-state conditions are achieved during microdialysis of minimal volumes of body fluids. RESULTS: A coaxial microdialysis probe with a length of 5 cm and an inner diameter of 200 μm is used for microdialysis of 10 μL of body fluid collected into a sampling fused silica capillary with an inner diameter 430 μm. Microdialysis is performed into 0.01 M HCl as a perfusate at stopped flow and 2 μL of the resulting microdialysate are subjected to analysis by capillary electrophoresis with contactless conductivity detection. Microdialysis pre-treatment is verified by analysis of 11 common amino acids at a 100 μM concentration level, resulting in recoveries of 98.3-102.5%. The electrophoretic separation of amino acids is performed in 8.5 M acetic acid at pH 1.37 as a background electrolyte with analysis time up to 4.5 min and LOD in the range of 0.12-0.28 μM. The reproducibility of the developed technique determined for the peak area ranges from 1.2 to 4.5%. Applicability is tested in the quantification of valine and leucine in plasma during fasting and subsequent reconvalescence. SIGNIFICANCE: The fabrication of a coaxial microdialysis probe for the laboratory preparation of microliter volumes of various types of clinical samples is described, which is coupled off-line with capillary electrophoretic monitoring of amino acids in 2 μL volumes of microdialysate. The developed methodology is suitable for quantification of 20 amino acids in whole human blood, plasma, tears and has potential for analysis of dry blood spots captured on hollow fibre.

• MeSH
• aminokyseliny * MeSH
• elektroforéza kapilární MeSH
• lidé MeSH
• mikrodialýza MeSH
• oxid křemičitý MeSH
• reprodukovatelnost výsledků MeSH
• tělesné tekutiny * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

A miniature probe for electromembrane extraction is developed and constructed. The tubular probe with an internal volume of 1.1 μL is made of polypropylene hollow fiber with a supported liquid membrane of 85% nitrophenyloctyl ether (NPOE) with 15% bis(2-ethylhexyl)phosphonic acid (DEHP). The probe is connected on-line to the electrophoresis with short separation capillary via an air assisted flow gating interface cast from poly (dimethylsiloxane). The compact instrument is computer controlled via LabView. The probe parameters are tested for extraction of creatinine and basic amino acids from artificial solution and human urine. The sensitivity of the electrophoretic determination after 300 s extraction at 150 V compared to the sensitivity without extraction is 4.9-fold and 2.6-fold higher for creatinine and arginine, respectively. The RSDs for peak area measured from 5 repeated extractions of 50 μM solutions are 7.5%, 7.2%, 8.6% and 9.2% for Crea, Lys, Arg and His, respectively. The probe can be used for all-day measurements. The preparation of the probe is simple and requires no special tool.

• MeSH
• elektroforéza kapilární * metody   MeSH
• ethery * chemie   MeSH
• kreatinin MeSH
• lidé MeSH
• membrány umělé MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

This article documents the beginning of the intellectual companionship between the founder of ethnomethodology, Harold Garfinkel, and Edward Rose, who is most often associated with his program of "ethno-inquiries." I present results from archival research focusing on the contacts and collaborations between Rose and Garfinkel in the years 1955-1965. First, I describe the review process for Rose and Felton's paper, submitted to the American Sociological Review in 1955, which Garfinkel reviewed and after Rose's rebuttal recommended for publication. The paper induced Garfinkel to write an extensive commentary that has remained unpublished. Second, I discuss the 1958 New Mexico conference sponsored by the Air Force, which was an opportunity for Rose and Garfinkel to work together on topics related to common-sense knowledge and scientific knowledge. Third, I give an overview of the ethnomethodological conferences in 1962 and 1963, supported by an Air Force grant written collaboratively by Rose and Garfinkel. Here I focus primarily on Rose's research on "small languages," which stimulated many discussions among the early ethnomethodologists. Rose's work and exchanges with Garfinkel demonstrate the former's affinity for miniaturization as a research approach and search for ways to empiricize topics of sociological theory in locally observable settings.

• MeSH
• altruismus MeSH
• archivy MeSH
• jazyk (prostředek komunikace) * MeSH
• kulturní antropologie MeSH
• lidé MeSH
• sociální teorie MeSH
• sociologie * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Spojené státy americké MeSH

The treatment of cartilage defects in trauma injuries and degenerative diseases represents a challenge for orthopedists. Advanced mesenchymal stromal cell (MSC)-based therapies are currently of interest for the repair of damaged cartilage. However, an approved system for MSC delivery and maintenance in the defect is still missing. This study aimed to evaluate the effect of autologous porcine bone marrow MSCs anchored in a commercially available polyglycolic acid-hyaluronan scaffold (Chondrotissue®) using autologous blood plasma-based hydrogel in the repair of osteochondral defects in a large animal model. The osteochondral defects were induced in twenty-four minipigs with terminated skeletal growth. Eight animals were left untreated, eight were treated with Chondrotissue® and eight received Chondrotissue® loaded with MSCs. The animals were terminated 90 days after surgery. Macroscopically, the untreated defects were filled with newly formed tissue to a greater extent than in the other groups. The histological evaluations showed that the defects treated with Chondrotissue® and Chondrotissue® loaded with pBMSCs contained a higher amount of hyaline cartilage and a lower amount of connective tissue, while untreated defects contained a higher amount of connective tissue and a lower amount of hyaline cartilage. In addition, undifferentiated connective tissue was observed at the edges of defects receiving Chondrotissue® loaded with MSCs, which may indicate the extracellular matrix production by transplanted MSCs. The immunological analysis of the blood samples revealed no immune response activation by MSCs application. This study demonstrated the successful and safe immobilization of MSCs in commercially available scaffolds and defect sites for cartilage defect repair.

• MeSH
• hydrogely MeSH
• kloubní chrupavka * chirurgie   MeSH
• krevní plazma MeSH
• mezenchymální kmenové buňky * fyziologie   MeSH
• miniaturní prasata MeSH
• modely u zvířat MeSH
• prasata MeSH
• tkáňové inženýrství MeSH
• transplantace mezenchymálních kmenových buněk * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH