The nerve agents of the A-series are relatively recent chemical weapons with no antidote available yet. Once inside the human body, those chemicals act similarly to the classic nerve agents, by binding to the catalytic residue Serine 203 (Ser203) of human acetylcholinesterase (HssAChE) and thus preventing the proper function of this enzyme. However, there is no experimental evidence yet if the current antidotes for intoxication by nerve agents are also capable of restoring AChE inhibited by the nerve agents of the A-series. In order to launch some light on this issue, we used computational techniques (molecular docking, molecular dynamics and MM-PBSA interaction energy calculations) to assess the performances of the four currently available commercial oximes (2-PAM, HI-6, obidoxime and trimedoxime) when in contact with HssAChE inhibited by the agent A-242. Based on the near-attack conformation (NAC) criterion, our results suggest that the commercial oximes would have limited efficacy to reactivate the enzyme since they are not able to properly approach the adduct Ser203-A-242. Among those oximes, trimedoxime seems to be the most promising, since it showed lower values of energy in the MM-PBSA calculations, a higher stability inside the catalytic anionic center (CAS) of HssAChE, and was able to adopt a position closer to the NAC that could enable the reactivation mechanism.

• Klíčová slova
• A-242, A-series nerve agents, Commercial oximes, Near-attack conformation,
• MeSH
• acetylcholinesterasa metabolismus   MeSH
• antidota farmakologie   MeSH
• cholinesterasové inhibitory chemie   toxicita   MeSH
• lidé MeSH
• nervová bojová látka * toxicita   MeSH
• organofosfáty MeSH
• oximy chemie   farmakologie   MeSH
• pyridinové sloučeniny farmakologie   MeSH
• reaktivátory cholinesterázy * farmakologie   MeSH
• simulace molekulového dockingu MeSH
• trimedoxim farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• acetylcholinesterasa MeSH
• antidota MeSH
• cholinesterasové inhibitory MeSH
• nervová bojová látka * MeSH
• novichok MeSH Prohlížeč
• organofosfáty MeSH
• oximy MeSH
• pyridinové sloučeniny MeSH
• reaktivátory cholinesterázy * MeSH
• trimedoxim MeSH

The misuse of novichok agents in assassination attempts has been reported in the international media since 2018. These relatively new class of neurotoxic agents is claimed to be more toxic than the agents of the G and V series and so far, there is no report yet in literature about potential antidotes against them. To shed some light into this issue, we report here the design and synthesis of NTMGMP, a surrogate of A-242 and also the first surrogate of a novichok agent useful for experimental evaluation of antidotes. Furthermore, the efficiency of the current commercial oximes to reactivate NTMGMP-inhibited acetylcholinesterase (AChE) was evaluated. The Ellman test was used to confirm the complete inhibition of AChE, and to compare the subsequent rates of reactivation in vitro as well as to evaluate aging. In parallel, molecular docking, molecular dynamics and MM-PBSA studies were performed on a computational model of the human AChE (HssAChE)/NTMGMP complex to assess the reactivation performances of the commercial oximes in silico. Experimental and theoretical studies matched the exact hierarchy of efficiency and pointed to trimedoxime as the most promising commercial oxime for reactivation of AChE inhibited by A-242.

• Klíčová slova
• A-242, AChE, Commercial reactivators, Novichoks, Surrogates,
• MeSH
• acetylcholinesterasa MeSH
• antidota farmakologie   MeSH
• cholinesterasové inhibitory toxicita   MeSH
• lidé MeSH
• nervová bojová látka * toxicita   MeSH
• oximy farmakologie   MeSH
• reaktivátory cholinesterázy * farmakologie   MeSH
• simulace molekulového dockingu MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• acetylcholinesterasa MeSH
• antidota MeSH
• cholinesterasové inhibitory MeSH
• nervová bojová látka * MeSH
• oximy MeSH
• reaktivátory cholinesterázy * MeSH

Neurodevelopmental disorders (NDDs), including severe paediatric epilepsy, autism and intellectual disabilities are heterogeneous conditions in which clinical genetic testing can often identify a pathogenic variant. For many of them, genetic therapies will be tested in this or the coming years in clinical trials. In contrast to first-generation symptomatic treatments, the new disease-modifying precision medicines require a genetic test-informed diagnosis before a patient can be enrolled in a clinical trial. However, even in 2022, most identified genetic variants in NDD genes are 'variants of uncertain significance'. To safely enrol patients in precision medicine clinical trials, it is important to increase our knowledge about which regions in NDD-associated proteins can 'tolerate' missense variants and which ones are 'essential' and will cause a NDD when mutated. In addition, knowledge about functionally indispensable regions in the 3D structure context of proteins can also provide insights into the molecular mechanisms of disease variants. We developed a novel consensus approach that overlays evolutionary, and population based genomic scores to identify 3D essential sites (Essential3D) on protein structures. After extensive benchmarking of AlphaFold predicted and experimentally solved protein structures, we generated the currently largest expert curated protein structure set for 242 NDDs and identified 14 377 Essential3D sites across 189 gene disorders associated proteins. We demonstrate that the consensus annotation of Essential3D sites improves prioritization of disease mutations over single annotations. The identified Essential3D sites were enriched for functional features such as intermembrane regions or active sites and discovered key inter-molecule interactions in protein complexes that were otherwise not annotated. Using the currently largest autism, developmental disorders, and epilepsies exome sequencing studies including >360 000 NDD patients and population controls, we found that missense variants at Essential3D sites are 8-fold enriched in patients. In summary, we developed a comprehensive protein structure set for 242 NDDs and identified 14 377 Essential3D sites in these. All data are available at https://es-ndd.broadinstitute.org for interactive visual inspection to enhance variant interpretation and development of mechanistic hypotheses for 242 NDDs genes. The provided resources will enhance clinical variant interpretation and in silico drug target development for NDD-associated genes and encoded proteins.

• Klíčová slova
• bioinformatics, genetics, neurodevelopmental disorder,
• MeSH
• dítě MeSH
• genetické testování MeSH
• lidé MeSH
• mentální retardace * genetika   MeSH
• missense mutace MeSH
• mutace genetika   MeSH
• neurovývojové poruchy * genetika   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

The precise preoperative staging of colorectal cancer is fundamental for surgical strategy, incomplete staging means incomplete treatment and poor outcome. Large-scale clinical evaluations of predictive markers are currently in progress, including determination of their ability to predict response of patients to therapy for advanced disease and for adjuvant treatment. Lack of specificity and sensitivity preclude the use of all existing serum markers for the early detection of colorectal carcinoma. The aim of the study was to investigate the clinical significance of serum tumor markers and biological activity markers -- oncofetal tumormarker CEA, mucin tumormarkers CA19-9, CA242, proliferative tumor markers Thymidine kinase, soluble cytoceratines fragments TPS, TPA, adhesive molecules ICAM - 1, VCAM -1, IGF-1, and adipocytokinins Adiponectin, Leptin in patients with colorectal cancer before primary operation. The study included 142 patients between the ages of 35 - 89 years. Operated between November 2003 to March 2006. We have confirmed that CA19-9 is besides CEA an important marker in colorectal cancer. Comparing CA19-9 and CA242 in preoperative staging, CA242 is more specific. Statistical significant difference between early and metastatic stage of colorectal cancer was not confirmed in markers: ICAM-1, VCAM, adiponectin, leptin. Statistical significant difference between early and metastatic stage of colorectal cancer was confirmed in markers: CEA, CA19-9, CA242, TPS, TPA, TK, IGF-1. None of the used markers was able to distinguish stage II and III, in other words to identify patients with infiltration of lymph nodes. This fact is very important in our aspirations to find which marker from periferal blood could help to poit out patients in risk of lymphatic infiltration and to indicate these patients for adjuvant therapy. Combination of CEA and either CA19-9 or CA242 can be recommended for preoperative investigation. CA 242 in this study seems to have slightly better results in preoperative staging.

• MeSH
• antigen CA-19-9 krev   MeSH
• antigeny sacharidové asociované s nádorem krev   MeSH
• karcinoembryonální antigen krev   MeSH
• kolorektální nádory diagnóza   mortalita   patologie   MeSH
• lidé MeSH
• nádorové biomarkery krev   MeSH
• prognóza MeSH
• staging nádorů MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigen CA-19-9 MeSH
• antigeny sacharidové asociované s nádorem MeSH
• CA 242 antigen MeSH Prohlížeč
• karcinoembryonální antigen MeSH
• nádorové biomarkery MeSH

BACKGROUND: A laparoscopy-based scoring system was developed by Fagotti et al (Fagotti or Predictive Index value (PIV)score) based on the intraoperative presence or absence of carcinomatosis on predefined sites. Later, the authors updated the PIV score calculated only in the absence of one or both absolute criteria of nonresectability (mesenteric retraction and miliary carcinomatosis of the small bowel) (updated PIV model). OBJECTIVE: The aim was to demonstrate the noninferiority of ultrasound to other imaging methods (contrast enhanced computed tomography (CT) and whole-body diffusion-weighted magnetic resonance imaging (WB-DWI)/MRI) in predicting nonresectable tumor (defined as residual disease >1 cm) using the updated PIV model in patients with tubo-ovarian cancer. The agreement between imaging and intraoperative findings as a reference was also calculated. STUDY DESIGN: This was a European prospective multicenter observational study. We included patients with suspected tubo-ovarian carcinoma who underwent preoperative staging and prediction of nonresectability at ultrasound, CT, WB-DWI/MRI, and surgical exploration. Ultrasound and CT were mandatory index tests, while WB-DWI/MRI was an optional test (non-available in all centers). The predictors of nonresectability were suspicious mesenteric retraction and/or miliary carcinomatosis of the small bowel or if absent, a PIV >8 (updated PIV model). The PIV score ranges from 0 to 12 according to the presence of disease in 6 predefined intra-abdominal sites (great omentum, liver surface, lesser omentum/stomach/spleen, parietal peritoneum, diaphragms, bowel serosa/mesentery). The reference standard was surgical outcome, in terms of residual disease >1 cm, assessed by laparoscopy and/or laparotomy. The area under the receiver operating characteristic curve (AUC) to assess the performance of the methods in predicting nonresectability was reported. Concordance between index tests at the detection of disease at 6 predefined sites and intraoperative exploration as reference standard was also calculated using Cohen's kappa. RESULTS: The study was between 2018 and 2022 in 5 European gynecological oncology centers. Data from 242 patients having both mandatory index tests (ultrasound and CT) were analyzed. 145/242 (59.9%) patients had no macroscopic residual tumor after surgery (R0) (5/145 laparoscopy and 140/145 laparotomy) and 17/242 (7.0%) had residual tumor ≤1 cm (R1) (laparotomy). In 80/242 patients (33.1%), the residual tumor was>1 cm (R2), 30 of them underwent laparotomy and maximum surgery was carried out, and 50/80 underwent laparoscopy only, because cytoreduction was not feasible in all of them. After excluding 18/242 (7.4%) patients operated on but not eligible for extensive surgery, the predictive performance of 3 imaging methods was analyzed in 167 women. The AUCs of all methods in discriminating between resectable and nonresectable tumor was 0.80 for ultrasound, 0.76 for CT, 0.71 for WB-DWI/MRI, and 0.90 for surgical exploration. Ultrasound had the highest agreement (Cohen's kappa ranging from 0.59 to 0.79) than CT and WB-DWI/MRI to assess all parameters included in the updated PIV model. CONCLUSION: Ultrasound showed noninferiority to CT and to WB-DWI/MRI in discriminating between resectable and nonresectable tumor using the updated PIV model. Ultrasound had the best agreement between imaging and intraoperative findings in the assessment of parameters included in the updated PIV model. Ultrasound is an acceptable method to assess abdominal disease and predict nonresectability in patients with tubo-ovarian cancer in the hands of specially trained ultrasound examiners.

• Klíčová slova
• computed tomography, laparoscopy, laparotomy, magnetic resonance, ovarian cancer, staging, ultrasonography,
• MeSH
• difuzní magnetická rezonance metody   MeSH
• dospělí MeSH
• laparoskopie metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory vaječníků * diagnostické zobrazování   chirurgie   patologie   MeSH
• počítačová rentgenová tomografie * MeSH
• prediktivní hodnota testů * MeSH
• prospektivní studie MeSH
• reziduální nádor diagnostické zobrazování   MeSH
• senioři MeSH
• staging nádorů MeSH
• ultrasonografie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• pozorovací studie MeSH

This study aimed to estimate the number of infants who died of unrecognized congenital adrenal hyperplasia (CAH) in Austria and the Czech Republic within the past 13 years, before the introduction of adequate neonatal screening. The study was based on retrospective analysis of neonatal screening cards of 242 infants who died suddenly between 7 days and 12 months of age and whose cause of death could not be identified. 17-hydroxyprogesterone (17-OHP) was measured by fluoroimmunoassay and positive samples were subsequently genotyped. Three infants out of 242 may have had unrecognized CAH due to CYP21 (steroid 21-hydroxylase) gene defect. Their newborn 17-OHP levels and CYP21 genotypes were 706 nmol/l and del/conv//del/conv, 53 nmol/l and I2//I2, and 811 nmol/l and I2//Gln318stop, respectively. CAH due to CYP21 defect can lead to sudden unexpected death without prior symptoms typical for the condition. Hence, newborn screening would have prevented these deaths had it been available. In addition, we have shown that the I2 point mutation that is expected to lead to simple virilizing form may lead to a fatal outcome.

• MeSH
• kojenec MeSH
• kongenitální adrenální hyperplazie epidemiologie   MeSH
• lidé MeSH
• náhlá smrt kojenců epidemiologie   MeSH
• novorozenec MeSH
• novorozenecký screening MeSH
• prevalence MeSH
• Check Tag
• kojenec MeSH
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Rakousko epidemiologie   MeSH

The effects of some antirheumatics on the formation and retraction of collagen lattices seeded with fibroblasts have been studied. Among the antirheumatics, diclofenac was the most active inhibitor of lattice retraction, then tropesin and to a lesser extent indomethacin. Ibuprofen which is known as a very slight inhibitor of protein synthesis was able to significantly enhance lattice retraction when 10 micrograms/ml (48.5 microM) and 50 micrograms/ml (242 microM) were used.

• MeSH
• antiflogistika nesteroidní farmakologie   MeSH
• fibroblasty chemie   MeSH
• kolagen metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antiflogistika nesteroidní MeSH
• kolagen MeSH

BACKGROUND: Activating point mutation of the BRAF gene, the most common genetic alteration reported in papillary thyroid carcinomas (PTC), has been associated with poor prognostic characteristics. AIM: Our objective was to determine the frequency of BRAFV600E mutation in PTC tumor tissues from the period 1960-2007 and to correlate it with clinicopathological parameters. SUBJECTS AND METHODS: DNAs were extracted from 242 PTCs, 23 sporadic medullary carcinomas, one anaplastic carcinoma and 6 poorly differentiated carcinomas. The presence of BRAFV600E mutation was determined using single strand conformation polymorphism method and verified by direct sequencing. RESULTS: BRAFV600E mutation was detected in 81 of 242 PTCs (33.5%), in one of 6 poorly differentiated carcinomas (16.7%) and in anaplastic carcinoma. BRAFV600E mutation was much less frequent in the follicular variant compared to classical variant and mixed follicular- classical variant of PTCs (p=0.001). BRAFV600E mutation was significantly associated with presence of nodal metastasis (p=0.029), more advanced TNM stage (p=0.014) and recurrence of disease (p=0.008). The mutation correlated with a higher age at diagnosis (p=0.049) and with a greater tumor size (p=0.041). Multivariate analysis confirmed these findings. The prevalence of BRAFV600E mutation before 1986 was significantly lower than after it (p=0.008). CONCLUSIONS: Our data suggest that BRAFV600E mutation is associated with high-risk clinicopathological characteristics of PTC and worse prognosis of patients. The frequency of the mutation significantly varied during the observed period but rather because of the different age distribution of patients in particular periods than as a consequence of Chernobyl accident.

• MeSH
• černobylská havárie MeSH
• DNA nádorová biosyntéza   genetika   MeSH
• dospělí MeSH
• exony genetika   MeSH
• frekvence genu MeSH
• invazivní růst nádoru genetika   MeSH
• kodon genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mutace fyziologie   MeSH
• nádory štítné žlázy epidemiologie   genetika   patologie   MeSH
• papilární karcinom epidemiologie   genetika   patologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• polymorfismus konformace jednovláknové DNA genetika   MeSH
• protoonkogenní proteiny B-Raf genetika   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• věkové faktory MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• BRAF protein, human MeSH Prohlížeč
• DNA nádorová MeSH
• kodon MeSH
• protoonkogenní proteiny B-Raf MeSH

Cryptosporidium parvum is one of the major causes of neonatal calf diarrhoea resulting in reduced farm productivity and compromised animal welfare worldwide. Livestock act as a major reservoir of this parasite, which can be transmitted to humans directly and/or indirectly, posing a public health risk. Research reports on the prevalence of Cryptosporidium in ruminants from east Mediterranean countries, including Cyprus, are limited. This study is the first to explore the occurrence of Cryptosporidium spp. in cattle up to 24 months old on the island of Cyprus. A total of 242 faecal samples were collected from 10 dairy cattle farms in Cyprus, all of which were screened for Cryptosporidium spp. using nested-PCR amplification targeting the small subunit of the ribosomal RNA (18S rRNA) gene. The 60 kDa glycoprotein (gp60) gene was also sequenced for the samples identified as Cryptosporidium parvum-positive to determine the subtypes present. The occurrence of Cryptosporidium was 43.8% (106/242) with at least one positive isolate in each farm sampled. Cryptosporidium bovis, Cryptosporidium ryanae and C. parvum were the only species identified, while the prevalence per farm ranged from 20-64%. Amongst these, the latter was the predominant species, representing 51.8% of all positive samples, followed by C. bovis (21.7%) and C. ryanae (31.1%). Five C. parvum subtypes were identified, four of which are zoonotic-IIaA14G1R1, IIaA15G1R1, IIaA15G2R1 and IIaA18G2R1. IIaA14G1R1 was the most abundant, representing 48.2% of all C. parvum positive samples, and was also the most widespread. This is the first report of zoonotic subtypes of C. parvum circulating in Cyprus. These results highlight the need for further research into the parasite focusing on its diversity, prevalence, host range and transmission dynamics on the island.

• Klíčová slova
• 18S rRNA, Cryptosporidium, Cryptosporidium parvum detection, Cyprus, calves, gp60, subtyping, zoonosis,
• Publikační typ
• časopisecké články MeSH

Limited data regarding the susceptibility of Actinobacillus pleuropneumoniae to antimicrobials has been published during recent years. Accordingly, the aim of the present study was to investigate the distribution of MICs for the isolates of A. pleuropneumoniae from diseased pigs in the Czech Republic between 2007 and 2009. A total of 242 isolates were tested for susceptibility to 16 antimicrobial agents by a broth microdilution method. A low degree of resistance was observed for florfenicol (0.8%), amoxicillin and clavulanic acid (0.8%), tilmicosin (1.2%), tiamulin (1.7%) and ampicillin (3.3%), whereas resistance to tetracycline was detected more frequently, 23.9% of isolates. Interestingly, resistance to florfenicol has not yet been reported in any study investigating antimicrobial resistance of A. pleuropneumoniae. By PCR the presence of the floR gene was confirmed in all florfenicol resistant isolates.

• MeSH
• Actinobacillus pleuropneumoniae účinky léků   izolace a purifikace   MeSH
• amoxicilin farmakologie   MeSH
• ampicilin farmakologie   MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální geny MeSH
• diterpeny farmakologie   MeSH
• kyselina klavulanová farmakologie   MeSH
• mikrobiální testy citlivosti MeSH
• nemoci prasat epidemiologie   mikrobiologie   MeSH
• prasata mikrobiologie   MeSH
• tetracyklin farmakologie   MeSH
• thiamfenikol analogy a deriváty   farmakologie   MeSH
• tylosin analogy a deriváty   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• amoxicilin MeSH
• ampicilin MeSH
• antibakteriální látky MeSH
• diterpeny MeSH
• florfenicol MeSH Prohlížeč
• kyselina klavulanová MeSH
• tetracyklin MeSH
• thiamfenikol MeSH
• tiamulin MeSH Prohlížeč
• tilmicosin MeSH Prohlížeč
• tylosin MeSH