The Colorado potato beetle (CPB; Leptinotarsa decemlineata) is an important potato pest with known resistance to pyrethroids and organophosphates in Czechia. Decreased efficacy of neonicotinoids has been observed in last decade. After the restriction of using chlorpyrifos, thiacloprid and thiamethoxam by EU regulation, growers seek for information about the resistance of CPB to used insecticides and recommended antiresistant strategies. The development of CPB resistance to selected insecticides was evaluated in bioassays in 69 local populations from Czechia in 2017-2022 and in 2007-2022 in small plot experiments in Zabcice in South Moravia. The mortality in each subpopulation in the bioassays was evaluated at the field-recommended rates of insecticides to estimate the 50% and 90% lethal concentrations (LC50 and LC90, respectively). High levels of CPB resistance to lambda-cyhalothrin and chlorpyrifos were demonstrated throughout Czechia, without significant changes between years and regions. The average mortality after application of the field-recommended rate of lambda-cyhalothrin was influenced by temperature before larvae were sampled for bioassays and decreased with increasing temperature in June. Downwards trends in the LC90 values of chlorpyrifos and the average mortality after application of the field-recommended rate of acetamiprid in the bioassay were recorded over a 6-year period. The baseline LC50 value (with 95% confidence limit) of 0.04 mg/L of chlorantraniliprole was established for Czech populations of CPBs for the purpose of resistance monitoring in the next years. Widespread resistance to pyrethroids, organophosphates and neonicotinoids was demonstrated, and changes in anti-resistant strategies to control CPBs were discussed.

• MeSH
• brouci * účinky léků   MeSH
• dursban * farmakologie   MeSH
• insekticidy * farmakologie   MeSH
• larva účinky léků   MeSH
• neonikotinoidy * farmakologie   MeSH
• nitrily farmakologie   MeSH
• pyrethriny farmakologie   MeSH
• rezistence k insekticidům * MeSH
• Solanum tuberosum parazitologie   MeSH
• thiamethoxam MeSH
• thiaziny * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• cyhalothrin MeSH Prohlížeč
• dursban * MeSH
• insekticidy * MeSH
• neonikotinoidy * MeSH
• nitrily MeSH
• pyrethriny MeSH
• thiacloprid MeSH Prohlížeč
• thiamethoxam MeSH
• thiaziny * MeSH

BACKGROUND: In spite of their importance as arthropod predators, spiders have received little attention in the risk assessment of pesticides. In addition, research has mainly focused on a few species commonly found in agricultural habitats. Spiders living in more natural ecosystems may also be exposed to and affected by pesticides, including insecticides. However, their sensitivity and factors driving possible variations in sensitivity between spider taxa are largely unknown. To fill this gap, we quantified the sensitivity of 28 spider species from a wide range of European ecosystems to lambda-cyhalothrin in an acute exposure scenario. RESULTS: Sensitivity varied among the tested populations by a factor of 30. Strong differences in sensitivity were observed between families, but also between genera within the Lycosidae. Apart from the variation explained by the phylogeny, spiders from boreal and polar climates were more sensitive than spiders from warmer areas. Overall, the median lethal concentration (LC50 ) of 85% of species was below the recommended application rate of lambda-cyhalothrin (75 ng a.i. cm-2 ). CONCLUSION: Our study underlines the high sensitivity of spiders to lambda-cyhalothrin, which can lead to unintended negative effects on pest suppression in areas treated with this insecticide. The strong differences observed between families and genera indicate that the functional composition of spider communities would change in affected areas. Overall, the variation in spider sensitivity suggests that multispecies investigations should be more widely considered in pesticide risk assessment. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

• Klíčová slova
• acute toxicity, lambda-cyhalothrin, species sensitivity distributions, spiders,
• MeSH
• ekosystém MeSH
• fylogeneze MeSH
• insekticidy * farmakologie   MeSH
• jedovatá zvířata * MeSH
• lidé MeSH
• můry * MeSH
• nitrily farmakologie   MeSH
• pavouci * MeSH
• pesticidy * farmakologie   MeSH
• pyrethriny * farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cyhalothrin MeSH Prohlížeč
• insekticidy * MeSH
• nitrily MeSH
• pesticidy * MeSH
• pyrethriny * MeSH

Androgen and glucocorticoid receptors have been recently described as key players in processes related to prostate cancer and mainly androgen receptor's inactivation was shown as an effective way for the prostate cancer treatment. Unfortunately, androgen deprivation therapy usually loses its effectivity and the disease frequently progresses into castration-resistant prostate cancer with poor prognosis. The role of the glucocorticoid receptor is associated with the mechanism of resistance; therefore, pharmacological targeting of glucocorticoid receptor in combination with antiandrogen treatment was shown as an alternative approach in the prostate cancer treatment. We introduce here the synthesis of novel 17α- and/or 21-ester or carbamate derivatives of hydrocortisone and evaluation of their biological activity towards androgen and glucocorticoid receptors in different prostate cancer cell lines. A 17α-butyryloxy-21-(alkyl)carbamoyloxy derivative 14 was found to diminish the transcriptional activity of both receptors (in single-digit micromolar range), with comparable potency to enzalutamide towards the androgen receptor, but weaker potency compared to mifepristone towards the glucocorticoid receptor. Lead compound inhibited proliferation and the formation of cell colonies in both androgen and glucocortiocid receptors-positive prostate cancer cell lines in low micromolar concentrations. Candidate compound 14 showed to interact with both receptors in cells and inhibited the translocation of receptors to nucleus and their activation phoshorylation. Moreover, binding to receptor's ligand binding domains was assessed by molecular modelling. Lead compound also induced the accumulation of cells in G1 phase and its combination with enzalutamide was shown to be more effective than enzalutamide alone.

• Klíčová slova
• Androgen receptor, Castration resistant prostate cancer, Glucocorticoid receptor, Hydrocortisone derivatives, Prostate cancer, Transcriptional activity,
• MeSH
• androgenní receptory * genetika   chemie   MeSH
• androgeny farmakologie   MeSH
• antagonisté androgenů farmakologie   MeSH
• chemorezistence MeSH
• hydrokortison farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prostaty rezistentní na kastraci * farmakoterapie   metabolismus   MeSH
• nitrily farmakologie   MeSH
• receptory glukokortikoidů MeSH
• signální transdukce MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• androgenní receptory * MeSH
• androgeny MeSH
• antagonisté androgenů MeSH
• enzalutamide MeSH Prohlížeč
• hydrokortison MeSH
• nitrily MeSH
• receptory glukokortikoidů MeSH

Cathepsin K (CatK) is a target for the treatment of osteoporosis, arthritis, and bone metastasis. Peptidomimetics with a cyanohydrazide warhead represent a new class of highly potent CatK inhibitors; however, their binding mechanism is unknown. We investigated two model cyanohydrazide inhibitors with differently positioned warheads: an azadipeptide nitrile Gü1303 and a 3-cyano-3-aza-β-amino acid Gü2602. Crystal structures of their covalent complexes were determined with mature CatK as well as a zymogen-like activation intermediate of CatK. Binding mode analysis, together with quantum chemical calculations, revealed that the extraordinary picomolar potency of Gü2602 is entropically favoured by its conformational flexibility at the nonprimed-primed subsites boundary. Furthermore, we demonstrated by live cell imaging that cyanohydrazides effectively target mature CatK in osteosarcoma cells. Cyanohydrazides also suppressed the maturation of CatK by inhibiting the autoactivation of the CatK zymogen. Our results provide structural insights for the rational design of cyanohydrazide inhibitors of CatK as potential drugs.

• Klíčová slova
• Cathepsin K, azadipeptide nitrile, cyanohydrazide warhead, protease inhibitor, structure,
• MeSH
• hydraziny chemie   farmakologie   MeSH
• inhibitory proteas chemie   farmakologie   MeSH
• kathepsin K antagonisté a inhibitory   metabolismus   MeSH
• lidé MeSH
• molekulární modely MeSH
• molekulární struktura MeSH
• nádorové buňky kultivované MeSH
• nitrily chemie   farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• CTSK protein, human MeSH Prohlížeč
• hydraziny MeSH
• inhibitory proteas MeSH
• kathepsin K MeSH
• nitrily MeSH

IMPORTANCE: To our knowledge, the Oral Ponesimod Versus Teriflunomide In Relapsing Multiple Sclerosis (OPTIMUM) trial is the first phase 3 study comparing 2 oral disease-modifying therapies for relapsing multiple sclerosis (RMS). OBJECTIVE: To compare the efficacy of ponesimod, a selective sphingosine-1-phosphate receptor 1 (S1P1) modulator with teriflunomide, a pyrimidine synthesis inhibitor, approved for the treatment of patients with RMS. DESIGN, SETTING, AND PARTICIPANTS: This multicenter, double-blind, active-comparator, superiority randomized clinical trial enrolled patients from April 27, 2015, to May 16, 2019, who were aged 18 to 55 years and had been diagnosed with multiple sclerosis per 2010 McDonald criteria, with a relapsing course from the onset, Expanded Disability Status Scale (EDSS) scores of 0 to 5.5, and recent clinical or magnetic resonance imaging disease activity. INTERVENTIONS: Patients were randomized (1:1) to 20 mg of ponesimod or 14 mg of teriflunomide once daily and the placebo for 108 weeks, with a 14-day gradual up-titration of ponesimod starting at 2 mg to mitigate first-dose cardiac effects of S1P1 modulators and a follow-up period of 30 days. MAIN OUTCOMES AND MEASURES: The primary end point was the annualized relapse rate. The secondary end points were the changes in symptom domain of Fatigue Symptom and Impact Questionnaire-Relapsing Multiple Sclerosis (FSIQ-RMS) at week 108, the number of combined unique active lesions per year on magnetic resonance imaging, and time to 12-week and 24-week confirmed disability accumulation. Safety and tolerability were assessed. Exploratory end points included the percentage change in brain volume and no evidence of disease activity (NEDA-3 and NEDA-4) status. RESULTS: For 1133 patients (567 receiving ponesimod and 566 receiving teriflunomide; median [range], 37.0 [18-55] years; 735 women [64.9%]), the relative rate reduction for ponesimod vs teriflunomide in the annualized relapse rate was 30.5% (0.202 vs 0.290; P < .001); the mean difference in FSIQ-RMS, -3.57 (-0.01 vs 3.56; P < .001); the relative risk reduction in combined unique active lesions per year, 56% (1.405 vs 3.164; P < .001); and the reduction in time to 12-week and 24-week confirmed disability accumulation risk estimates, 17% (10.1% vs 12.4%; P = .29) and 16% (8.1% vs 9.9; P = .37), respectively. Brain volume loss at week 108 was lower by 0.34% (-0.91% vs -1.25%; P < .001); the odds ratio for NEDA-3 achievement was 1.70 (25.0% vs 16.4%; P < .001). Incidence of treatment-emergent adverse events (502 of 565 [88.8%] vs 499 of 566 [88.2%]) and serious treatment-emergent adverse events (49 [8.7%] vs 46 [8.1%]) was similar for both groups. Treatment discontinuations because of adverse events was more common in the ponesimod group (49 of 565 [8.7%] vs 34 of 566 [6.0%]). CONCLUSIONS AND RELEVANCE: In this study, ponesimod was superior to teriflunomide on annualized relapse rate reduction, fatigue, magnetic resonance imaging activity, brain volume loss, and no evidence of disease activity status, but not confirmed disability accumulation. The safety profile was in line with the previous safety observations with ponesimod and the known profile of other S1P receptor modulators. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02425644.

• MeSH
• dospělí MeSH
• hydroxybutyráty farmakologie   MeSH
• imunologické faktory terapeutické užití   MeSH
• krotonáty farmakologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lokální recidiva nádoru farmakoterapie   MeSH
• magnetická rezonanční tomografie metody   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• nitrily farmakologie   MeSH
• progrese nemoci MeSH
• relabující-remitující roztroušená skleróza farmakoterapie   MeSH
• thiazoly farmakologie   MeSH
• toluidiny farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• randomizované kontrolované studie MeSH
• srovnávací studie MeSH
• Názvy látek
• hydroxybutyráty MeSH
• imunologické faktory MeSH
• krotonáty MeSH
• nitrily MeSH
• ponesimod MeSH Prohlížeč
• teriflunomide MeSH Prohlížeč
• thiazoly MeSH
• toluidiny MeSH

Prostate cancer (PC) is a potentially high-risk disease and the most common cancer in American men. It is a leading cause of cancer-related deaths in men in the US, second only to lung and bronchus cancer. Advanced and metastatic PC is initially treated with androgen deprivation therapy (ADT), but nearly all cases eventually progress to castrate-resistant prostate cancer (CRPC). CRPC is incurable in the metastatic stage but can be slowed by some conventional chemotherapeutics and second-generation ADT, such as enzalutamide and abiraterone. Therefore, novel therapeutic strategies are urgently needed. Prostate-specific membrane antigen (PSMA) is overexpressed in almost all aggressive PCs. PSMA is widely used as a target for PC imaging and drug delivery. Anti-PSMA monoclonal antibodies (mAbs) have been developed as bioligands for diagnostic imaging and targeted PC therapy. However, these mAbs are successfully used in PC imaging and only a few have gone beyond phase-I for targeted therapy. The 5D3 mAb is a novel, high-affinity, and fast-internalizing anti-PSMA antibody. Importantly, 5D3 mAb demonstrates a unique pattern of cellular localization to the centrosome after internalization in PSMA(+) PC3-PIP cells. These characteristics make 5D3 mAb an ideal bioligand to deliver tubulin inhibitors, such as mertansine, to the cell centrosome, leading to mitotic arrest and elimination of dividing PC cells. We have successfully developed a 5D3 mAb- and mertansine (DM1)-based antibody-drug conjugate (ADC) and evaluated it in vitro for binding affinity, internalization, and cytotoxicity. The in vivo therapeutic efficacy of 5D3-DM1 ADC was evaluated in PSMA(+) PC3-PIP and PSMA(-) PC3-Flu mouse models of human PC. This therapeutic study has revealed that this new anti-PSMA ADC can successfully control the growth of PSMA(+) tumors without inducing systemic toxicity.

• Klíčová slova
• 5D3 antibody, MCC linker, anti-PSMA antibody, antibody-drug conjugates (ADC), drug delivery, mertansine (DM1), prostate cancer, prostate-specific membrane antigen (PSMA), targeted therapy,
• MeSH
• androsteny farmakologie   MeSH
• antagonisté androgenů farmakologie   MeSH
• antigeny povrchové metabolismus   MeSH
• benzamidy farmakologie   MeSH
• buňky PC-3 MeSH
• centrozom metabolismus   MeSH
• fenylthiohydantoin farmakologie   MeSH
• glutamátkarboxypeptidasa II metabolismus   MeSH
• imunokonjugáty farmakologie   MeSH
• lidé MeSH
• modulátory tubulinu farmakologie   MeSH
• monoklonální protilátky farmakologie   MeSH
• myši nahé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory prostaty rezistentní na kastraci farmakoterapie   metabolismus   MeSH
• nitrily farmakologie   MeSH
• xenogenní modely - testy antitumorózní aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• abiraterone MeSH Prohlížeč
• androsteny MeSH
• antagonisté androgenů MeSH
• antigeny povrchové MeSH
• benzamidy MeSH
• enzalutamide MeSH Prohlížeč
• fenylthiohydantoin MeSH
• FOLH1 protein, human MeSH Prohlížeč
• glutamátkarboxypeptidasa II MeSH
• imunokonjugáty MeSH
• modulátory tubulinu MeSH
• monoklonální protilátky MeSH
• nitrily MeSH

Prostate cancer (PCa) is the most common cancer and the second leading cause of cancer-related deaths of men in Western countries. Androgen deprivation therapy is initially successful, however eventually fails, and tumors progress to the more aggressive castration-resistant PCa (CRPC). Yet, androgen receptor (AR) usually remains as a major regulator of tumor cell proliferation in CRPC. Interleukin-23 (IL-23) was recently shown to promote the development of CRPC by driving AR transcription. Here we used the androgen-sensitive LNCaP, castration-resistant C4-2, and 22Rv1 cells. Interestingly, cellular senescence is induced in these human cell lines by treatment with the AR antagonists enzalutamide (ENZ) or darolutamide (ODM), which might be one underlying mechanism for inhibition of PCa cell proliferation. Treatment with IL-23 alone did not change cellular senescence levels in these cell lines, whereas IL-23 inhibited significantly cellular senescence levels induced by ENZ or ODM in both CRPC cell lines C4-2 and 22Rv1 but not in LNCaP cells. This indicates a response of IL-23 specific in CRPC cells. Generating LNCaP and C4-2 three-dimensional (3D) spheroids and treatment with AR antagonists resulted in the reduced spheroid volume and thus growth inhibition. However, the combination of AR antagonists with IL-23 did not affect the antagonist-mediated reduction of spheroid volumes. This observation was confirmed with proliferation assays using adherent monolayer cell cultures. Taken together, the data indicate that IL-23 treatment reduces the AR antagonists-induced level of cellular senescence of CRPC cells, which could be one possible mechanism for promoting castration resistance.

• Klíčová slova
• Androgen receptor antagonists, Cellular senescence, Interleukin-23, Prostate cancer spheroids,
• MeSH
• antagonisté androgenních receptorů farmakologie   terapeutické užití   MeSH
• benzamidy farmakologie   terapeutické užití   MeSH
• fenylthiohydantoin farmakologie   terapeutické užití   MeSH
• interleukin-23 metabolismus   MeSH
• lidé MeSH
• nitrily farmakologie   terapeutické užití   MeSH
• protokoly antitumorózní kombinované chemoterapie farmakologie   terapeutické užití   MeSH
• pyrazoly farmakologie   terapeutické užití   MeSH
• stárnutí buněk MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antagonisté androgenních receptorů MeSH
• benzamidy MeSH
• darolutamide MeSH Prohlížeč
• enzalutamide MeSH Prohlížeč
• fenylthiohydantoin MeSH
• interleukin-23 MeSH
• nitrily MeSH
• pyrazoly MeSH

BACKGROUND: Androgen receptor targeted therapies have emerged as an effective tool to manage advanced prostate cancer (PCa). Nevertheless, frequent occurrence of therapy resistance represents a major challenge in the clinical management of patients, also because the molecular mechanisms behind therapy resistance are not yet fully understood. In the present study, we therefore aimed to identify novel targets to intervene with therapy resistance using gene expression analysis of PCa co-culture spheroids where PCa cells are grown in the presence of cancer-associated fibroblasts (CAFs) and which have been previously shown to be a reliable model for antiandrogen resistance. METHODS: Gene expression changes of co-culture spheroids (LNCaP and DuCaP seeded together with CAFs) were identified by Illumina microarray profiling. Real-time PCR, Western blotting, immunohistochemistry and cell viability assays in 2D and 3D culture were performed to validate the expression of selected targets in vitro and in vivo. Cytokine profiling was conducted to analyze CAF-conditioned medium. RESULTS: Gene expression analysis of co-culture spheroids revealed that CAFs induced a significant upregulation of cholesterol and steroid biosynthesis pathways in PCa cells. Cytokine profiling revealed high amounts of pro-inflammatory, pro-migratory and pro-angiogenic factors in the CAF supernatant. In particular, two genes, 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2 (HMGCS2) and aldo-keto reductase family 1 member C3 (AKR1C3), were significantly upregulated in PCa cells upon co-culture with CAFs. Both enzymes were also significantly increased in human PCa compared to benign tissue with AKR1C3 expression even being associated with Gleason score and metastatic status. Inhibiting HMGCS2 and AKR1C3 resulted in significant growth retardation of co-culture spheroids as well as of various castration and enzalutamide resistant cell lines in 2D and 3D culture, underscoring their putative role in PCa. Importantly, dual targeting of cholesterol and steroid biosynthesis with simvastatin, a commonly prescribed cholesterol synthesis inhibitor, and an inhibitor against AKR1C3 had the strongest growth inhibitory effect. CONCLUSIONS: From our results we conclude that CAFs induce an upregulation of cholesterol and steroid biosynthesis in PCa cells, driving them into AR targeted therapy resistance. Blocking both pathways with simvastatin and an AKR1C3 inhibitor may therefore be a promising approach to overcome resistances to AR targeted therapies in PCa. Video abstract.

• Klíčová slova
• AKR1C3, Antiandrogens, Castration resistance, Cholesterol, HMGCS2, Prostate cancer, Simvastatin, Steroid metabolism,
• MeSH
• androgenní receptory metabolismus   MeSH
• anotace sekvence MeSH
• benzamidy farmakologie   MeSH
• biologické modely MeSH
• biosyntetické dráhy genetika   MeSH
• buněčné sféroidy metabolismus   patologie   MeSH
• buněčný cyklus genetika   MeSH
• chemorezistence účinky léků   genetika   MeSH
• cholesterol biosyntéza   MeSH
• extracelulární matrix metabolismus   MeSH
• fenotyp MeSH
• fenylthiohydantoin farmakologie   MeSH
• fibroblasty asociované s nádorem metabolismus   patologie   MeSH
• kultivační média speciální farmakologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prostaty rezistentní na kastraci genetika   patologie   MeSH
• nádory prostaty genetika   metabolismus   patologie   MeSH
• nitrily farmakologie   MeSH
• progrese nemoci * MeSH
• proliferace buněk genetika   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• senioři MeSH
• simvastatin farmakologie   MeSH
• stanovení celkové genové exprese MeSH
• upregulace * MeSH
• viabilita buněk účinky léků   genetika   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• androgenní receptory MeSH
• AR protein, human MeSH Prohlížeč
• benzamidy MeSH
• cholesterol MeSH
• enzalutamide MeSH Prohlížeč
• fenylthiohydantoin MeSH
• kultivační média speciální MeSH
• nitrily MeSH
• simvastatin MeSH

Tuberous sclerosis complex (TSC) is an autosomal dominantly inherited neurocutaneous disorder caused by inactivating mutations in TSC1 or TSC2, key regulators of the mechanistic target of rapamycin complex 1 (mTORC1) pathway. In the CNS, TSC is characterized by cortical tubers, subependymal nodules and subependymal giant cell astrocytomas (SEGAs). SEGAs may lead to impaired circulation of CSF resulting in hydrocephalus and raised intracranial pressure in patients with TSC. Currently, surgical resection and mTORC1 inhibitors are the recommended treatment options for patients with SEGA. In the present study, high-throughput RNA-sequencing (SEGAs n = 19, periventricular control n = 8) was used in combination with computational approaches to unravel the complexity of SEGA development. We identified 9400 mRNAs and 94 microRNAs differentially expressed in SEGAs compared to control tissue. The SEGA transcriptome profile was enriched for the mitogen-activated protein kinase (MAPK) pathway, a major regulator of cell proliferation and survival. Analysis at the protein level confirmed that extracellular signal-regulated kinase (ERK) is activated in SEGAs. Subsequently, the inhibition of ERK independently of mTORC1 blockade decreased efficiently the proliferation of primary patient-derived SEGA cultures. Furthermore, we found that LAMTOR1, LAMTOR2, LAMTOR3, LAMTOR4 and LAMTOR5 were overexpressed at both gene and protein levels in SEGA compared to control tissue. Taken together LAMTOR1-5 can form a complex, known as the 'Ragulator' complex, which is known to activate both mTORC1 and MAPK/ERK pathways. Overall, this study shows that the MAPK/ERK pathway could be used as a target for treatment independent of, or in combination with mTORC1 inhibitors for TSC patients. Moreover, our study provides initial evidence of a possible link between the constitutive activated mTORC1 pathway and a secondary driver pathway of tumour growth.

• Klíčová slova
• SEGA, TSC, low grade glioma, sequencing,
• MeSH
• adaptorové proteiny signální transdukční genetika   metabolismus   MeSH
• astrocytom etiologie   genetika   metabolismus   MeSH
• astrocyty účinky léků   metabolismus   MeSH
• butadieny farmakologie   MeSH
• dítě MeSH
• dospělí MeSH
• extracelulárním signálem regulované MAP kinasy antagonisté a inhibitory   genetika   metabolismus   MeSH
• hamartin genetika   MeSH
• inhibitory enzymů farmakologie   MeSH
• intracelulární signální peptidy a proteiny genetika   metabolismus   MeSH
• kojenec MeSH
• lidé MeSH
• MAP kinasový signální systém genetika   MeSH
• mechanistické cílové místo rapamycinového komplexu 1 MeSH
• messenger RNA metabolismus   MeSH
• mikro RNA metabolismus   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• nádorové buňky kultivované MeSH
• nádory mozku komplikace   genetika   metabolismus   MeSH
• nitrily farmakologie   MeSH
• předškolní dítě MeSH
• sekvenční analýza RNA MeSH
• sekvenování transkriptomu MeSH
• stanovení celkové genové exprese MeSH
• tuberin genetika   MeSH
• tuberózní skleróza komplikace   genetika   MeSH
• výměnné faktory guaninnukleotidů genetika   metabolismus   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec 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
• adaptorové proteiny signální transdukční MeSH
• butadieny MeSH
• extracelulárním signálem regulované MAP kinasy MeSH
• hamartin MeSH
• inhibitory enzymů MeSH
• intracelulární signální peptidy a proteiny MeSH
• LAMTOR1 protein, human MeSH Prohlížeč
• LAMTOR3 protein, human MeSH Prohlížeč
• LAMTOR4 protein, human MeSH Prohlížeč
• LAMTOR5 protein, human MeSH Prohlížeč
• mechanistické cílové místo rapamycinového komplexu 1 MeSH
• messenger RNA MeSH
• mikro RNA MeSH
• nitrily MeSH
• ragulator complex protein LAMTOR2, human MeSH Prohlížeč
• TSC1 protein, human MeSH Prohlížeč
• TSC2 protein, human MeSH Prohlížeč
• tuberin MeSH
• U 0126 MeSH Prohlížeč
• výměnné faktory guaninnukleotidů MeSH

Varroa destructor is the major cause of honey bee (Apis mellifera) colony losses. Mite control is limited to several miticides. The overuse of tau-fluvalinate has resulted in resistance via a knockdown resistance (kdr) mutation in the sodium channel gene NaVChs (L925V/I/M). In this study, we used the discriminating concentration of tau-fluvalinate (0.25 µg/mL) to detect the resistance of mites in a bioassay. Further, we verified the presence of the kdr mutation in mites from the bioassay via PCR amplification of a fragment of the voltage-gated sodium channel gene (NaVCh), restriction fragment length polymorphisms (RFLPs), and densitometry analyses in pools of surviving or dead mites. Resistance values corresponding to the densitometry of the resistant allele were related to mite survival. In the vial test, the survival of the control group was significantly higher (70.4%) than that of the tau-fluvalinate-treated group (34.3%). Mite survival in the vial test was significantly correlated with the mean proportion of resistance values. Individuals that died after tau-fluvalinate application exhibited an average resistance value of 0.0783, whereas individuals that survived exhibited an average resistance of 0.400. The concentration of tau-fluvalinate in the vials was checked using high performance liquid chromatography under different temperatures and exposure times, and indicates that the stability of tau-fluvalinate stored in the refrigerator (4 ± 1 °C) is at least 14 days. PCR-RFLP of the NaVCh gene fragment verified that the vial test is a suitable, rapid, and cost-effective method for the identification of tau-fluvalinate resistance based on kdr mutation in V. destructor in apiaries.

• Klíčová slova
• Acaricide, Apiary, Honey bee, Restriction, Varroa destructor,
• MeSH
• akaricidy farmakologie   MeSH
• biotest metody   MeSH
• denzitometrie metody   MeSH
• léková rezistence genetika   MeSH
• nitrily farmakologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• pyrethriny farmakologie   MeSH
• Varroidae účinky léků   genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• akaricidy MeSH
• fluvalinate MeSH Prohlížeč
• nitrily MeSH
• pyrethriny MeSH