Exposure to loud sounds damages the auditory periphery and induces maladaptive changes in central parts of the auditory system. Diminished peripheral afferentation and altered inhibition influence the processing of sounds in the auditory cortex. It is unclear, however, which types of inhibitory interneurons are affected by acoustic trauma. Here we used single-unit electrophysiological recording and two-photon calcium imaging in anesthetized mice to evaluate the effects of acute acoustic trauma (125 dB SPL, white noise, 5 min) on the response properties of neurons in the core auditory cortex. Electrophysiological measurements suggested the selective impact of acoustic trauma on inhibitory interneurons in the auditory cortex. To further investigate which interneuronal types were affected, we used two-photon calcium imaging to record the activity of neurons in cortical layers 2/3 and 4, specifically focusing on parvalbumin-positive (PV+) and somatostatin-positive (SST+) interneurons. Spontaneous and pure-tone-evoked firing rates of SST+ interneurons increased in layer 4 immediately after acoustic trauma and remained almost unchanged in layer 2/3. Furthermore, PV+ interneurons with high best frequencies increased their evoked-to-spontaneous firing rate ratios only in layer 2/3 and did not change in layer 4. Finally, acoustic trauma unmasked low-frequency excitatory inputs only in layer 2/3. Our results demonstrate layer-specific changes in the activity of auditory cortical inhibitory interneurons within minutes after acoustic trauma.

• MeSH
• akční potenciály MeSH
• interneurony metabolismus   fyziologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nedoslýchavost z hluku patofyziologie   MeSH
• parvalbuminy genetika   metabolismus   MeSH
• sluchové evokované potenciály * MeSH
• sluchové korové centrum cytologie   patofyziologie   MeSH
• somatostatin genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In the mammalian neocortex, the calcium-binding protein calretinin is expressed in a subset of cortical interneurons. In the recent years, research on interneurons is one of the most rapidly growing fields in neuroscience. This review summarizes the actual knowledge of the functions of calretinin in neuronal homeostasis and particularly of the distribution, connectivity and physiological properties of calretinin expressing interneurons in the neocortex of rodents and primates, including humans. The possible neuroprotective role of calretinin and the presumed “resistance” of calretinin-expressing interneurons to various pathological processes are also discussed.

• MeSH
• financování organizované MeSH
• homeostáza fyziologie   MeSH
• lidé MeSH
• neokortex cytologie   fyziologie   MeSH
• neurony fyziologie   MeSH
• S100 kalcium vázající protein G fyziologie   MeSH
• savci MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• přehledy MeSH

• MeSH
• biotransformace MeSH
• chromatografie na tenké vrstvě metody   MeSH
• cyklin-dependentní kinasy antagonisté a inhibitory   farmakologie   MeSH
• hodnocení léčiv MeSH
• inhibitory cytochromu P450 MeSH
• jaterní mikrozomy metabolismus   MeSH
• myši MeSH
• oxidace-redukce MeSH
• systém (enzymů) cytochromů P-450 MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• srovnávací studie MeSH

• MeSH
• analýza moči MeSH
• chromatografie na tenké vrstvě MeSH
• denzitometrie MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• poruchy spojené s užíváním psychoaktivních látek MeSH
• závislost na morfiu diagnóza   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

The purpose of this study was to determine the role of lipotoxicity in vascular smooth muscle (VSM). C1-BODIPY 500/510 C12 used to assess the ability of VSM A7r5 cells to transport long-chain fatty acids showed that lipid transport did not appear to limit metabolism. Thin layer chromatography revealed that storage of transported fatty acid occurred primarily as mono- and diglycerides and fatty acids but not as triglycerides. We used lipid-induced apoptosis as a measure of lipotoxicity and found that 1.5 mM palmitate (6.8:1) bound to albumin resulted in a 15-fold increase in the number of apoptotic cells compared to the control at 24 hours. This apoptosis did not seem to be due to an increase in reactive oxygen species (ROS) since VSM cells incubated in palmitate showed less ROS production than cells incubated in albumin only. Similar exposure to oleate did not significantly increase the number of apoptotic cells compared to the control. Oleate actually significantly attenuated the apoptosis induced by palmitate, suggesting that unsaturated fatty acids have a protective effect on cells undergoing palmitate-induced apoptosis. These results suggest that vascular smooth muscle is vulnerable to lipotoxicity and that this lipotoxicity may play a role in the development of atherosclerosis.

• MeSH
• apoptóza genetika   MeSH
• ateroskleróza etiologie   patofyziologie   MeSH
• chromatografie na tenké vrstvě metody   využití   MeSH
• finanční podpora výzkumu jako téma MeSH
• kyseliny olejové chemie   MeSH
• mastné kyseliny chemie   krev   MeSH
• palmitany chemie   MeSH
• poruchy metabolismu lipidů diagnóza   etiologie   MeSH
• reaktivní formy kyslíku chemie   metabolismus   škodlivé účinky   MeSH
• svaly hladké cévní cytologie   fyziologie   patologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

BACKGROUND: The non-viral production of CAR-T cells through electroporation of transposon DNA plasmids is an alternative approach to lentiviral/retroviral methods. This method is particularly suitable for early-phase clinical trials involving novel types of CAR-T cells. The primary disadvantage of non-viral methods is the lower production efficiency compared to viral-based methods, which becomes a limiting factor for CAR-T production, especially in chemotherapy-pretreated lymphopenic patients. METHODS: We describe a good manufacturing practice (GMP)-compliant protocol for producing CD19 and CD123-specific CAR-T cells based on the electroporation of transposon vectors. The lymphocytes were purified from the blood of patients undergoing chemotherapy for B-NHL or AML and were electroporated with piggyBac transposon encoding CAR19 or CAR123, respectively. Electroporated cells were then polyclonally activated by anti-CD3/CD28 antibodies and a combination of cytokines (IL-4, IL-7, IL-21). The expansion was carried out in the presence of irradiated allogeneic blood-derived mononuclear cells (i.e., the feeder) for up to 21 days. RESULTS: Expansion in the presence of the feeder enhanced CAR-T production yield (4.5-fold in CAR19 and 9.3-fold in CAR123). Detailed flow-cytometric analysis revealed the persistence of early-memory CAR-T cells and a low vector-copy number after production in the presence of the feeder, with no negative impact on the cytotoxicity of feeder-produced CAR19 and CAR123 T cells. Furthermore, large-scale manufacturing of CAR19 carried out under GMP conditions using PBMCs obtained from B-NHL patients (starting number=200x10e6 cells) enabled the production of >50x10e6 CAR19 in 7 out of 8 cases in the presence of the feeder while only in 2 out of 8 cases without the feeder. CONCLUSIONS: The described approach enables GMP-compatible production of sufficient numbers of CAR19 and CAR123 T cells for clinical application and provides the basis for non-viral manufacturing of novel experimental CAR-T cells that can be tested in early-phase clinical trials. This manufacturing approach can complement and advance novel experimental immunotherapeutic strategies against human hematologic malignancies.

• MeSH
• akutní myeloidní leukemie * terapie   imunologie   genetika   MeSH
• allogeneické buňky imunologie   MeSH
• antigeny CD19 * imunologie   genetika   MeSH
• B-buněčný lymfom terapie   imunologie   genetika   MeSH
• chimerické antigenní receptory * genetika   imunologie   MeSH
• elektroporace MeSH
• imunoterapie adoptivní * metody   MeSH
• lidé MeSH
• podkladové buňky MeSH
• T-lymfocyty imunologie   metabolismus   MeSH
• transpozibilní elementy DNA * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Domoic acid (DA) is a potent marine neurotoxine present in seafood. Intoxication by DA causes gastrointestinal symptoms like vomiting and diarrhoea and also the so-called amnesic shellfish poisoning (inflicting memory impairment and seizures). Since exposure to non-convulsive doses is relevant to the human health, we investigated the effect of low dose DA administration in adult Wistar rats. Rats were administered with DA at the dose 1.0 mg/kg and their behavior was monitored for one hour in three sessions. The first session started immediately after DA administration. The second and third session started one and two weeks later. After the third session, the histochemical analysis of the hippocampi of the animals was conducted (Fluoro-Jade B, bis-benzimide). DA increased time spent by locomotion and distance travelled in the second half of the first session and this effect was pronounced during the second and third session. Exploratory rearing was decreased by DA administration in the first half of the first session. DA influenced the grooming in biphasic manner (decrease followed by an increase of time spent by grooming). This biphasic trend was observed even two weeks after the DA administration. Histochemistry of DA treated rats did not confirm the presence of apoptotic bodies, Fluoro-Jade B positive cells were not found neither in CA1 nor CA3 area of the hippocampi. Our study revealed that a low dose of DA affect short and long-term the spontaneous behavior of rats without inducing neuronal damage.

• MeSH
• apoptóza účinky léků   MeSH
• depolarizující myorelaxancia aplikace a dávkování   toxicita   MeSH
• hipokampální oblast CA1 cytologie   účinky léků   MeSH
• hipokampální oblast CA3 cytologie   účinky léků   MeSH
• kyselina kainová aplikace a dávkování   analogy a deriváty   toxicita   MeSH
• lokomoce účinky léků   MeSH
• náhodné rozdělení MeSH
• péče o zevnějšek u zvířat účinky léků   MeSH
• pohybová aktivita účinky léků   MeSH
• potkani Wistar MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• apoptóza imunologie   MeSH
• buněčná smrt genetika   MeSH
• choriové klky imunologie   růst a vývoj   MeSH
• diagnostické techniky molekulární metody   MeSH
• endometrium cytologie   imunologie   růst a vývoj   MeSH
• imunohistochemie metody   MeSH
• lidé MeSH
• nádory endometria genetika   imunologie   MeSH
• průtoková cytometrie MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH

Nález kyseliny ll-nor-delta-9-tetrahydrocannabinol-9-karboxylové v moči je považován za spolehlivý důkaz předchozí konzumace cannabinoidů, protože vzniká v organismu při biotransformaci delta-9-tetrahydrocannabinolu, hlavní účinné složky produktů z konopí a hašiše. Účelem předkládané práce bylo njgít optimální postup při izolaci kyseliny ll-nor-delta-9-tetrahydrocannabinol-9karboxylové z moče a další podmínky, za kterých je možná její identifikace metodou chromatografie na tenké vrstvě po předběžném pozitivním screeningu cannabinoidů imunochemickou metodou EMIT.

ll-nor-delta-9-tetrahydrocannabinol-9-carboxylic acid is the biotransformation product of delta-9tetrahydrocannabinol, that is the main active compound of the product from cannabis: marihuana and hashish. Its identification in urine is supposed to be the best indication of previous cannabis consuption. The aim of this study was to find conditions for the use of thin layer chromatography as an identification method of ll-nor-delta-9-tetrahydrocannabinol-9-carboxylic in urine after positive preliminary screening of cannabinoids by EMIT method.

• MeSH
• abúzus marihuany moč   MeSH
• chromatografie na tenké vrstvě MeSH
• kanabinoidy moč   MeSH
• kanabinol metabolismus   MeSH
• kyseliny karboxylové moč   MeSH
• lidé MeSH
• moč chemie   MeSH
• Check Tag
• lidé MeSH

Ecstasy or MDMA as a psychoactive drug and hallucinogen is considered one of the most commonly used drugs in the world. This psychotropic substance is discussed both as sexually stimulating and reducing fear and anxiety. Amphetamines also destroy neurons in some brain areas. The aim of this study was to investigate the effects of MDMA on anxiety and apoptosis of hippocampal neurons. Forty-two male Wistar rats of mean weight 200-220 g were used and distributed into six groups [control, control-saline, and experimental groups (1.25, 2.5, 5, 10 mg/kg)]. Rats in experimental groups received MDMA at different doses for seven days by intraperitoneal injection and the control-saline group received saline (1 ml/kg); anxiety was then investigated by plus-maze test. Forty-eight hours after behavioural testing brains were taken from animals and fixed, and after tissue processing, slices were stained with TUNEL kit for apoptotic cells. The area densities of apoptotic neurons were measured throughout the hippocampus and compared in all groups (P < 0.05). Physiological studies showed that 1.25 mg/kg and 2.5 mg/kg doses caused anti-anxiety behaviour and 5 and 10 mg/kg doses of MDMA caused anxietylike behaviour. Moreover, our histological study showed that ecstasy increased apoptotic cell numbers and the highest increase was observed with the 10 mg/kg dose of MDMA. We concluded that MDMA can cause different responses of anxiety-like behaviour in different doses. This phenomenon causes a different ratio of apoptosis in hippocampal formation. Reduction of anxiety-like behaviour induced by the 2.5 mg/kg dose of MDMA can control apoptosis.

• MeSH
• apoptóza * MeSH
• chování zvířat MeSH
• gyrus dentatus patologie   MeSH
• hipokampální oblast CA1 patologie   MeSH
• hipokampální oblast CA3 patologie   MeSH
• hipokampus patologie   MeSH
• koncové značení zlomů DNA in situ MeSH
• N-methyl-3,4-methylendioxyamfetamin MeSH
• neurony patologie   MeSH
• počet buněk MeSH
• potkani Wistar MeSH
• úzkost chemicky indukované   patologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH