Feruloyl esterases (FAEs) are a crucial component of the hemicellulose-degrading enzyme family that facilitates the degradation of lignocellulose while releasing hydroxycinnamic acids such as ferulic acid with high added value. Currently, the low enzyme yield of FAEs is one of the primary factors limiting its application. Therefore, in this paper, we optimized the fermentation conditions for the expression of FAE BpFaeT132C-D143C with excellent thermal stability in Escherichia coli by experimental design. Firstly, we explored the effects of 11 factors such as medium type, isopropyl-β-D-thiogalactopyranoside (IPTG) concentration, and inoculum size on BpFaeT132C-D143C activity separately by the single factor design. Then, the significance of the effects of seven factors, such as post-induction temperature, shaker rotational speed, and inoculum size on BpFaeT132C-D143C activity, was analyzed by Plackett-Burman design. We identified the main factors affecting the fermentation conditions of E. coli expressing BpFaeT132C-D143C as post-induction temperature, pre-induction period, and post-induction period. Finally, we used the steepest ascent path design and response surface method to optimize the levels of these three factors further. Under the optimal conditions, the activity of BpFaeT132C-D143C was 3.58 U/ml, which was a significant 6.6-fold increase compared to the pre-optimization (0.47 U/ml), demonstrating the effectiveness of this optimization process. Moreover, BpFaeT132C-D143C activity was 1.52 U/ml in a 3-l fermenter under the abovementioned optimal conditions. It was determined that the expression of BpFaeT132C-D143C in E. coli was predominantly intracellular in the cytoplasm. This study lays the foundation for further research on BpFaeT132C-D143C in degrading agricultural waste transformation applications.

• MeSH
• Escherichia coli * genetika   metabolismus   enzymologie   MeSH
• fermentace * MeSH
• isopropylthiogalaktosid metabolismus   MeSH
• karboxylesterhydrolasy * genetika   metabolismus   chemie   biosyntéza   MeSH
• kultivační média chemie   MeSH
• kyseliny kumarové metabolismus   MeSH
• lignin MeSH
• rekombinantní proteiny genetika   metabolismus   biosyntéza   chemie   MeSH
• stabilita enzymů MeSH
• teplota MeSH
• Publikační typ
• časopisecké články MeSH

Angioedém je otok zahrnující podkožní a/nebo submukózní vrstvy tkáně, které postihují obličej, rty, krk a končetiny, dutinu ústní, hrtan a/nebo střevní sliznici. Diferenciální diagnostika angioedémů je mezioborová. Z hlediska patofyziologie může být angioedém klasifikován jako angioedém zprostředkovaný histaminem a angioedém zprostředkovaný bradykininem. Histaminem zprostředkovaný angioedém je častější a souvisí s aktivací a degranulací žírných buněk a bazofilů, bývá provázen svědivou a zarudlou urtikárií. Angiedém zprostředkovaný bradykininem může zahrnovat formy hereditárního angioedému, získaného deficitu C1 inhibitoru a angioedémy spojené s inhibitorem angiotenzin-konvertujícího enzymu či dalších léků. Je charakterizován nadměrnou lokální tvorbou bradykininu se vznikem bolestivého angioedému, není spojen se svědivou kopřivkou, má delší trvání a často břišní příznaky. Je rezistentní vůči standardním terapiím, jako je adrenalin, glukokortikoidy a antihistaminika. V rámci diferenciální diagnostiky v akutní fázi je vhodné provést laboratorní vyšetření tryptázy k odlišení histaminového angioedému v souběhu anafylaxe a C4 složku komplementu jako screening pro bradykininový angioedém. Hereditární angioedém (HAE) je vzácné, geneticky podmíněné onemocnění s autozomálně dominantním přenosem a variabilním spektrem bradykininových angioedémů. V širším kontextu se jedná o imunodeficitní onemocnění, klasifikované na HAE s deficiencí C1 inhibitoru (HAE-C1-INH) a HAE s normální hladinou a funkcí C1 inhibitoru (HAE nC1-INH), s mutacemi jiného (mnohdy ještě neznámého) typu. Vznik center pro diagnostiku a péči o pacienty s HAE a získanými bradykininovými angioedémy (AAE) významně zlepšil životní osudy těchto pacientů. Do center jsou konzilárně odesíláni i pacienti s atypickými angioedémy (s převahou bradykininové etiologie).

Angioedema is swelling involving the subcutaneous and/or submucosal layers of tissue that affects the face, lips, neck and extremities, oral cavity, larynx, and/or intestinal mucosa. The differential diagnosis of angioedema is interdisciplinary. From a pathophysiological perspective, angioedema can be classified as histamine-mediated angioedema and bradykinin-mediated angioedema. Histamine-mediated angioedema is more common and is associated with activation and degranulation of mast cells and basophils, and is often accompanied by pruritic and erythematous urticaria. Bradykinin-mediated angioedema can include forms of hereditary angioedema, acquired C1 inhibitor deficiency, and angioedema associated with angiotensin-converting enzyme inhibitors or other drugs. It is characterized by excessive local production of bradykinin with the development of painful angioedema, is not associated with pruritic urticaria, has a longer duration, and often has abdominal symptoms. It is resistant to standard therapies such as adrenaline, glucocorticoids, and antihistamines. As part of the differential diagnosis in the acute phase, it is appropriate to perform a laboratory test for tryptase to differentiate histamine angioedema in conjunction with anaphylaxis and the C4 component of complement as a screening for bradykinin angioedema. Hereditary angioedema (HAE) is a rare, genetically determined disease with autosomal dominant transmission and a variable spectrum of bradykinin angioedema. In a broader context, it is an immunodeficiency disease, classified into HAE with C1 inhibitor deficiency (HAE-C1-INH) and HAE with normal levels and function of C1 inhibitor (HAE nC1-INH), with mutations of another (often still unknown) type. The establishment of centers for the diagnosis and care of patients with HAE and acquired bradykinin angioedema (AAE) has significantly improved the lives of these patients. Patients with atypical angioedema (with a predominance of bradykinin etiology) are also referred to the centers.

• MeSH
• angioedém etiologie   klasifikace   terapie   MeSH
• diferenciální diagnóza MeSH
• dítě MeSH
• hereditární angioedémy * diagnóza   genetika   patofyziologie   terapie   MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• věk při počátku nemoci MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• ženské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH
• přehledy MeSH

Garadacimab je nově registrovaný lék určený k profylaktické léčbě pacientů s hereditárním angioedémem s deficitem C1 inhibitoru. Blokuje aktivovaný koagulační faktor XII a tím inhibuje tvorbu bradykininu, který je hlavním mediátorem zodpovědným za vznik otoku u pacientů s touto diagnózou. Klinické studie naznačují jeho vysokou účinnost a bezpečnost.

Garadacimab is a newly approved drug designed for the prophylactic treatment of patients with hereditary angioedema due to C1 inhibitor deficiency. It blocks activated coagulation factor XII, thereby inhibiting the production of bradykinin, the key mediator responsible for the development of swelling in patients with this condition. Clinical studies suggest its high efficiency and safety.

• Klíčová slova
• garadacimab, profylaktická léčba,
• MeSH
• hereditární angioedémy * farmakoterapie   prevence a kontrola   MeSH
• humanizované monoklonální protilátky farmakologie   terapeutické užití   MeSH
• lidé MeSH
• randomizované kontrolované studie jako téma MeSH
• Check Tag
• lidé MeSH

Background: Hereditary angioedema (HAE) attacks substantially impair health-related quality of life (HRQoL). Current World Allergy Organization and the European Academy of Allergy and Clinical Immunology guidelines goals include complete control and normalization of patients' lives. Garadacimab (anti-activated factor XII monoclonal antibody) reduced the mean attack rate after first administration in the pivotal phase III (VANGUARD; NCT04656418) and ongoing long-term phase III open-label extension (OLE) (NCT04739059) studies. Objective: To report exploratory HRQoL data from the interim analysis of the phase III OLE study (data cutoff February 13, 2023). Methods: Patients ages ≥12 years and with HAE received garadacimab 200 mg subcutaneously once monthly in the OLE study. The patient population comprised patients who were garadacimab naive (received placebo in the previous phase III study and newly enrolled patients) and patients who received garadacimab in previous phase II/III studies. The Angioedema Quality of Life (AE-QoL) questionnaire, Treatment Satisfaction Questionnaire for Medication version II (TSQM II), and Work Productivity and Activity Impairment: General Health (WPAI:GH) questionnaire were administered at baseline and every 3 months during the OLE study. AE-QoL and TSQM II scores were evaluated in comparison with minimal clinically important differences (MCID). Results: Overall, 90 patients who were garadacimab naive and 71 patients with previous garadacimab exposure received garadacimab in the phase III OLE study. The mean ± standard deviation AE-QoL total score improved by 34.2 ± 18.8 points in patients who were garadacimab naive and by 2.3 ± 13.1 points further to the reduction experienced in patients with previous garadacimab exposure. The AE-QoL MCID was met by 92.1% of patients who were garadacimab naive; 81.6% of patients with previous garadacimab exposure experienced stable AE-QoL scores or further improvements per MCID. TSQM II scores were improved from day 1 with garadacimab and sustained to month 12. Improvements in WPAI:GH scores were consistent with AE-QoL and TSQM II. Conclusion: Garadacimab elicited clinically meaningful long-term improvements in HRQoL, work productivity, and treatment satisfaction in patients with HAE, which brought them closer to complete disease control and normalization of life.Clinical trial NCT04739059, clinicaltrials.gov.

• MeSH
• dospělí MeSH
• hereditární angioedémy * farmakoterapie   diagnóza   MeSH
• humanizované monoklonální protilátky * terapeutické užití   MeSH
• kvalita života * MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• průzkumy a dotazníky MeSH
• senioři MeSH
• spokojenost pacientů MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky, fáze III MeSH

The adaptive immune response critically hinges on the functionality of T cell receptors, governed by complex molecular mechanisms, including ubiquitination. In this study, we delved into the role of in T cell immunity, focusing on T cell-B cell conjugate formation and T cell activation. Using a CRISPR-Cas9 screening approach targeting deubiquitinases genes in Jurkat T cells, we identified BAP1 as a key positive regulator of T cell-B cell conjugate formation. Subsequent investigations into BAP1 knockout cells revealed impaired T cell activation, evidenced by decreased MAPK and NF-kB signaling pathways and reduced CD69 expression upon T cell receptor stimulation. Flow cytometry and qPCR analyses demonstrated that BAP1 deficiency leads to decreased surface expression of T cell receptor complex components and reduced mRNA levels of the co-stimulatory molecule CD28. Notably, the observed phenotypes associated with BAP1 knockout are specific to T cells and fully dependent on BAP1 catalytic activity. In-depth RNA-seq and mass spectrometry analyses further revealed that BAP1 deficiency induces broad mRNA and protein expression changes. Overall, our findings elucidate the vital role of BAP1 in T cell biology, especially in T cell-B cell conjugate formation and T cell activation, offering new insights and directions for future research in immune regulation.

• MeSH
• aktivace lymfocytů * imunologie   MeSH
• B-lymfocyty * imunologie   metabolismus   MeSH
• Jurkat buňky MeSH
• lidé MeSH
• nádorové supresorové proteiny * metabolismus   genetika   MeSH
• receptory antigenů T-buněk * metabolismus   MeSH
• signální transdukce MeSH
• T-lymfocyty * imunologie   metabolismus   MeSH
• thiolesterasa ubikvitinu * genetika   metabolismus   nedostatek   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Spontaneous tumor regression is a recognized phenomenon across various cancer types. Recent research emphasizes the alterations in autoantibodies against carbonic anhydrase I (CA I) (anti-CA I) levels as potential prognostic markers for various malignancies. Particularly, autoantibodies targeting CA I and II appear to induce cellular damage by inhibiting their respective protein's catalytic functions. Our study illuminates the profound impact of anti-CA I autoantibodies from patient serum on the esterase activity of human CA I, exhibiting inhibitory effects akin to the acetazolamide inhibitor. Concurrently, our newly synthesized mouse monoclonal IgG antibody, mAb 2B8, against human CA I showcased a potent inhibitory action. An in-depth exploration into mAb 2B8's binding dynamics with its target enzyme was undertaken. Leveraging epitope extraction and phage display library techniques, we identified the amino acid sequence DFWTYP (positions 191-196 of CA I) as crucial for mAb 2B8's interaction. In 3-D structural analysis, this sequence is spatially adjacent to a previously identified epitope (DFWTYP) that interacts with patient-derived autoantibodies. Critically, mAb 2B8 demonstrated an ability to infiltrate eukaryotic cells, engaging specifically with its intracytoplasmic target. This positions mAb 2B8 as a promising model for future studies aimed at tumor cell eradication.

• MeSH
• autoprotilátky * imunologie   metabolismus   MeSH
• epitopy imunologie   chemie   MeSH
• karboanhydrasa I * metabolismus   antagonisté a inhibitory   MeSH
• lidé MeSH
• monoklonální protilátky * imunologie   MeSH
• myši MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Background: Lipocalin-2 (LCN2) is a protein that has been associated with skeletal muscle regeneration, but details regarding its role in Arthritis remain unclear. The aim of the current study was to investigate LCN2 levels of Arthritis patients and its relationship with oxidative and antioxidative factors.Methods: The study includes (125) blood samples of persons aged 20-65 years were divided into a control group (apparently healthy) consisting of 55 samples [31female, 24 males] and a Patient group consisting of 70 samples [37female, 33 males] who were attending the bone diseases consultation unit at the Ibn Sina Teaching Hospital in Mosul, Iraq. Venous blood samples (10 ml) were collected after overnight fasting. To conduct Clinical analyses: Serum LCN2 level was determined by ELISA, also Malonaldehyde, glutathione, vitamin E, vitamin C, peroxy nitrite, peroxidase, and aryl esterase were estimated.Results: The findings revealed a significant increase in the levels of LCN2 in Arthritis compared to the control group and there was a significant decrease in the concentration of vitamin C, glutathione, vitamin E and the activity of the arylesterase in serum of patients with arthritis compared with the control group. Also, a significant increase in the activity of peroxidase, concentration of peroxynitrite and malondialdehyde for patients than a control group.Conclusion: These findings imply that LCN2 may play a substantial role in iron-related oxidative stress damage in arthritis. Thus a therapeutic candidate target for treatment.

• MeSH
• antioxidancia metabolismus   MeSH
• artritida * diagnóza   krev   metabolismus   MeSH
• glutathion krev   MeSH
• index tělesné hmotnosti MeSH
• lidé MeSH
• lipokalin-2 * analýza   krev   metabolismus   MeSH
• malondialdehyd krev   MeSH
• oxidační stres * fyziologie   MeSH
• vitamin E krev   MeSH
• Check Tag
• lidé MeSH

Hereditární angioedém (HAE) je vzácné, geneticky podmíněné onemocnění s autozomálně dominantním přenosem a variabilním spektrem klinických projevů i život ohrožujících. V širším kontextu se jedná o imunodeficitní onemocnění, klasifikované na HAE s deficiencí C1 inhibitoru (HAE-C1-INH, dříve HAE-I a HAE-II) a HAE s normální hladinou a funkcí C1 inhibitoru (HAE nC1-INH) označovaný též jako HAE-III typu, s mutacemi jiného (mnohdy ještě neznámého) typu. Klinickým projevem jsou masivní otoky podkoží a/nebo sliznic v důsledku nekontrolované aktivace komplementového a kininového systému. V důsledku lokální nadprodukce bradykininu vznikají typické angioedémy. Vzácněji může deficit C1-inhibitoru vzniknout s vazbou na jiné patologické stavy (autoimunitní, lymfoproliferace, monoklonální gamapatie) – jedná se o získaný angioedém (AAE, acquired angioedema). Za samostatný klinický syndrom, který je nutno odlišit od získaného angioedému, je považován ACE inhibitory indukovaný angioedém (AE-ACEi). Důležitá je farmakologická anamnéza, rizikové mohou být i sartany, inhibitory mTOR, gliptiny, mezi dalšími léky je uváděn aliskiren, sakubitril, tkáňový aktivátor plasminogenu. Vznik center pro diagnostiku a péči o pacienty s HAE/AAE významně zlepšil životní osudy těchto pacientů. Do center jsou konzilárně odesíláni i pacienti s atypickými angioedémy (s převahou bradykininové etiologie).

Hereditary angioedema (HAE) is a rare, genetically determined disease with autosomal dominant transmission and a variable spectrum of clinical and life-threatening manifestations. In a broader context, it is an immunodeficiency disease, classified into HAE with a deficiency of C1 inhibitor (HAE-C1-INH, formerly HAE-I and HAE-II) and HAE with a normal level and function of C1 inhibitor (HAE nC1-INH), also referred to as HAE-III type, with mutations of another (often still unknown) type. The clinical manifestation is massive swelling of the subcutaneous tissue and/or mucous membranes due to uncontrolled activation of the complement and kinin systems. Local overproduction of bradykinin results in typical angioedema. More rarely, C1-inhibitor deficiency can arise in connection with other pathological conditions (autoimmune, lymphoproliferation, monoclonal gammopathy) – this is acquired angioedema (AAE). Angioedema induced by ACE inhibitors (AE-ACEi) is considered a separate clinical syndrome that must be distinguished from acquired angioedema. The pharmacological anamnesis is important, sartans, mTOR inhibitors, gliptins can also be risky, aliskiren, sacubitril, tissue plasminogen activator are mentioned among other drugs. The emergence of centers for the diagnosis and care of patients with HAE/AAE significantly improved the lives of these patients. Patients with atypical angioedema (predominantly of bradykinin etiology) are also sent to the centers on a consular basis.

• MeSH
• bradykinin metabolismus   škodlivé účinky   MeSH
• diferenciální diagnóza MeSH
• hereditární angioedémy * diagnóza   farmakoterapie   klasifikace   MeSH
• lidé MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

A novel group of 5,6-dihydropyrido [2',1':2,3]imidazo [4,5-c]quinolines was prepared via a microwave assisted one-pot telescopic approach. The synthetic sequence involves the formation of an amine precursor of imidazo [1,2-a]pyridine via condensation and reduction under microwave irradiation. Subsequently, the Pictet-Spengler cyclisation reaction occurs with ketones (cyclic or acyclic) to obtain substituted 5,6-dihydropyrido [2',1':2,3]imidazo [4,5-c]quinolines in excellent yields. The compounds were tested as neuroprotective agents. Observed protection of neuron-like cells, SH-SY5Y differentiated with ATRA, in Parkinson's and Huntington's disease models inspired further mechanistic studies of protective activity against damage induced by 1-methyl-4-phenylpyridinium (MPP+), a compound causing Parkinson's disease. The novel compounds exhibit similar or higher potency than ebselen, an established drug with antioxidant activity, in the cells against MPP + -induced total cellular superoxide production and cell death. However, they exhibit a significantly higher capacity to reduce mitochondrial superoxide and preserve mitochondrial membrane potential. We also observed marked differences between a selected derivative and ebselen in terms of normalizing MPP + -induced phosphorylation of Akt and ERK1/2. The cytoprotective activity was abrogated when signaling through cannabinoid receptor CB2 was blocked. The compounds also inhibit both acetylcholine and butyrylcholine esterases. Overall the data show that novel 5,6-dihydropyrido [2',1':2,3]imidazo [4,5-c]quinoline have a broad cytoprotective activity which is mediated by several mechanisms including mitoprotection.

• MeSH
• chinoliny * farmakologie   chemie   chemická syntéza   MeSH
• cholinesterasové inhibitory * farmakologie   chemie   chemická syntéza   MeSH
• lidé MeSH
• molekulární struktura MeSH
• nádorové buněčné linie MeSH
• neuroprotektivní látky * farmakologie   chemie   chemická syntéza   MeSH
• receptor kanabinoidní CB2 * metabolismus   antagonisté a inhibitory   MeSH
• signální transdukce * účinky léků   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BAP1-inactivated melanocytic tumors represent a subset of epithelioid melanocytic neoplasms resulting from biallelic inactivation of the BAP1 gene and by a driver mutation that activate the MAP kinase pathway, most commonly BRAFV600E. They occur sporadically or, less common, in the setting of BAP1 tumor predisposition syndrome caused by a BAP1 germline mutation that predisposes to several malignancies including cutaneous and uveal melanoma. To date, only few cases of BAP1-inactivated melanomas have been reported. We present a case of a 35-year-old woman presented with a melanocytic lesion microscopically composed of 3 distinct melanocytic populations, suggesting a stepwise progression model to melanoma from a conventional nevus through a melanocytoma stage. This progression was also supported from a molecular viewpoint given BRAFV600E, BAP1, and TERT-p hot spot mutations detected by targeted mutational analysis. Four atypical melanocytic lesions were removed from the patient's back, and the same A BAP1 c.856A>T, p.(Lys286Ter) mutation was detected on either tumoral or normal tissue samples. To the best of our knowledge, this is the first case of BAP1-inactivated melanoma with a documented TERT-p hot spot mutation manifesting as the first presentation of BAP1 tumor predisposition syndrome.

• MeSH
• dědičné nádorové syndromy * patologie   MeSH
• dospělí MeSH
• epiteloidní a vřetenobuněčný névus * patologie   MeSH
• lidé MeSH
• melanocyty patologie   MeSH
• melanom * patologie   MeSH
• mutace MeSH
• nádorové supresorové proteiny genetika   metabolismus   MeSH
• nádory kůže * patologie   MeSH
• thiolesterasa ubikvitinu genetika   MeSH
• zárodečné mutace MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• přehledy MeSH