The development of canine immunotolerant monoclonal antibodies can accelerate the invention of new medicines for both canine and human diseases. We develop a methodology to clone the naive, somatically mutated variable domain repertoire from canine B cell mRNA using 5'RACE PCR. A set of degenerate primers were then designed and used to clone variable domain genes into archival "holding" plasmid libraries. These archived variable domain genes were then combinatorially ligated to produce a scFv M13 phage library. Next-generation long-read and short-read DNA sequencing methodologies were developed to annotate features of the cloned library including CDR diversity and IGHV/IGKV/IGLV subfamily distribution. A synthetic immunoglobulin G was developed from this scFv library to the canine immune checkpoint receptor PD-1. This synthetic platform can be used to clone and annotate archived antibody variable domain genes for use in perpetuity in order to develop improved preclinical models for the treatment of complex human diseases.

• MeSH
• antigeny CD279 imunologie   MeSH
• jednořetězcové protilátky * imunologie   genetika   MeSH
• lidé MeSH
• monoklonální protilátky imunologie   genetika   MeSH
• nádory imunologie   terapie   MeSH
• peptidová knihovna MeSH
• psi MeSH
• rekombinantní proteiny imunologie   genetika   MeSH
• translační biomedicínský výzkum MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Permeability is an important molecular property in drug discovery, as it co-determines pharmacokinetics whenever a drug crosses the phospholipid bilayer, e.g., into the cell, in the gastrointestinal tract, or across the blood-brain barrier. Many methods for the determination of permeability have been developed, including cell line assays (CACO-2 and MDCK), cell-free model systems like parallel artificial membrane permeability assay (PAMPA) mimicking, e.g., gastrointestinal epithelia or the skin, as well as the black lipid membrane (BLM) and submicrometer liposomes. Furthermore, many in silico approaches have been developed for permeability prediction: meta-analysis of publicly available databases for permeability data (MolMeDB and ChEMBL) was performed to establish their usability. Four experimental and two computational methods were evaluated. It was shown that repeatability of the reported permeability measurement is not great even for the same method. For the PAMPA method, two different permeabilities are reported: intrinsic and apparent. They can vary in degrees of magnitude; thus, we suggest being extra cautious using literature data on permeability. When we compared data for the same molecules using different methods, the best agreement was between cell-based methods and between BLM and computational methods. Existence of unstirred water layer (UWL) permeability limits the data agreement between cell-based methods (and apparent PAMPA) with data that are not limited by UWL permeability (computational methods, BLM, intrinsic PAMPA). Therefore, different methods have different limitations. Cell-based methods provide results only in a small range of permeabilities (-8 to -4 in cm/s), and computational methods can predict a wider range of permeabilities beyond physical limitations, but their precision is therefore limited. BLM with liposomes can be used for both fast and slow permeating molecules, but its usage is more complicated than standard transwell techniques. To sum up, when working with in-house measured or published permeability data, we recommend caution in interpreting and combining them.

• MeSH
• buňky MDCK MeSH
• Caco-2 buňky MeSH
• hematoencefalická bariéra metabolismus   MeSH
• lidé MeSH
• liposomy * chemie   MeSH
• membrány umělé MeSH
• permeabilita buněčné membrány fyziologie   MeSH
• permeabilita * MeSH
• psi MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• metaanalýza MeSH

The olfactory response of insect vectors such as phlebotomine sand flies is a key facet for investigating their interactions with vertebrate hosts and associated vector-borne pathogens. Such studies are mainly performed by assessing the electrophysiological response and the olfactory behaviour of these arthropods towards volatile organic compounds (VOCs) produced by hosts. Nonetheless, few studies are available for species of the subgenera Lutzomyia and Nyssomyia in South America, leaving a void for Old World sand fly species of the genus Phlebotomus. In this study, we evaluated the olfactory responses of Phlebotomus perniciosus, one of the most important vectors of Leishmania infantum in the Old World. To test the P. perniciosus behavioural response to VOCs, 28 compounds isolated from humans and dogs were assessed using electrophysiological (i.e., electroantennogram, EAG) and behavioural assays (i.e., Y-tube olfactometer). In the EAG trials, 14 compounds (i.e., acetic acid, nonanoic acid, 2-propanol, 2-butanol, pentanal, hexanal, nonanal, trans-2-nonenal, decanal, myrcene, p-cymene, verbenone, 2-ethyl-1-hexanol, and acetonitrile) elicited high antennal responses (i.e., ≥ 0.30 mV) in female sand flies, being those VOCs selected for the behavioural assays. From the 14 compounds tested in the Y-tube olfactometer, nonanal was significantly attractive for P. perniciosus females, whereas myrcene and p-cymene were significantly repellents (p < 0.05). The attraction indexes varied from 0.53 for nonanal (i.e., most attractive) to -0.47 to p-cymene (i.e., most repellent). Overall, our results shed light on the role of olfactory cues routing host seeking behaviour in P. perniciosus, with implications to develop sustainable sand fly monitoring as well as control in leishmaniasis endemic areas.

• MeSH
• chování zvířat účinky léků   MeSH
• hmyz - vektory fyziologie   účinky léků   MeSH
• Leishmania infantum účinky léků   fyziologie   MeSH
• lidé MeSH
• Phlebotomus * fyziologie   účinky léků   MeSH
• psi MeSH
• těkavé organické sloučeniny * farmakologie   chemie   analýza   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• psi MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Small nucleolar RNAs (snoRNAs) are non-coding RNAs (ncRNAs) that regulate many cellular processes. Changes in the profiles of cellular ncRNAs and those secreted in exosomes are observed during viral infection. In our study, we analysed differences in expression profiles of snoRNAs isolated from exosomes of influenza (IAV)-infected and non-infected MDCK cells using high-throughput sequencing. The analysis revealed 133 significantly differentially regulated snoRNAs (131 upregulated and 2 downregulated), including 93 SNORD, 38 SNORA, and 2 SCARNA. The most upregulated was SNORD58 (log2FoldChange = 9.61), while the only downregulated snoRNAs were SNORD3 (log2FC = -2.98) and SNORA74 (log2FC = -2.67). Several snoRNAs previously described as involved in viral infections were upregulated, including SNORD27, SNORD28, SNORD29, SNORD58, and SNORD44. In total, 533 interactors of dysregulated snoRNAs were identified using the RNAinter database with an assigned confidence score ≥ 0.25. The main groups of predicted interactors were transcription factors (TFs, 169 interactors) and RNA-binding proteins (RBPs, 130 interactors). Among the most important were pioneer TFs such as POU5F1, SOX2, CEBPB, and MYC, while in the RBP category, notable interactors included Polr2a, TNRC6A, IGF2BP3, and FMRP. Our results suggest that snoRNAs are involved in pro-viral activity, although follow-up studies including experimental validation would be beneficial.

• MeSH
• buňky MDCK MeSH
• exozómy * metabolismus   genetika   MeSH
• infekce viry z čeledi Orthomyxoviridae virologie   metabolismus   genetika   MeSH
• malá jadérková RNA * genetika   metabolismus   MeSH
• psi MeSH
• stanovení celkové genové exprese MeSH
• virus chřipky A * fyziologie   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVES: To evaluate the base excess response during acute in vivo carbon dioxide changes. DESIGN: Secondary analysis of individual participant data from experimental studies. SETTING: Three experimental studies investigating the effect of acute in vivo respiratory derangements on acid-base variables. SUBJECTS: Eighty-nine (canine and human) carbon dioxide exposures. INTERVENTIONS: Arterial carbon dioxide titration through environmental chambers or mechanical ventilation. MEASUREMENTS AND MAIN RESULTS: For each subject, base excess was calculated using bicarbonate and pH using a fixed buffer power of 16.2. Analyses were performed using linear regression with arterial dioxide (predictor), base excess (outcome), and studies (interaction term). All studies show different baselines and slopes for base excess across carbon dioxide titrations methods. Individual subjects show substantial, and potentially clinically relevant, variations in base excess response across the hypercapnic range. Using a mathematical simulation of 10,000 buffer power coefficients we determined that a coefficient of 12.1 (95% CI, 9.1-15.1) instead of 16.2 facilitates a more conceptually appropriate in vivo base excess equation for general clinical application. CONCLUSIONS: In vivo changes in carbon dioxide leads to changes in base excess that may be clinically relevant for individual patients. A buffer power coefficient of 16.2 may not be appropriate in vivo and needs external validation in a range of clinical settings.

• MeSH
• acidobazická rovnováha * fyziologie   MeSH
• dospělí MeSH
• hyperkapnie patofyziologie   metabolismus   MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• oxid uhličitý * metabolismus   MeSH
• poruchy acidobazické rovnováhy patofyziologie   metabolismus   MeSH
• psi MeSH
• umělé dýchání MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• psi MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Předkládané sdělení popisuje případ systémové infekce vyvolané Pasteurella multocida. Infekce byla prokázána u 79letého muže, který byl do nemocničního zařízení dovezen po pádu z gauče. Onemocnění se projevilo rozvojem febrilního stavu, zimnicí, bolestmi kloubů a pádem. Laboratorně byla zjištěna elevace CRP, mírné zvýšení dusíkatých metabolitů, v krevním obraze hraniční leukocytóza, trombocytopenie. Agens bylo prokázáno v hemokultuře a v kultivačním vyšetření ve stěru z rány. Pacient byl léčen v úvodu cefalosporinem III. generace (cefotaxime), doléčován Xorimaxem. Článek je doplněn informacemi o etiologickém agens, jeho historii a přehledem literatury dokumentovaných komplikovaných případů pasteurelózy.

POZOR! při kopírování abstrakt kontrolovat slova na konci řádků originálu!!!

• MeSH
• cefalosporiny třetí generace farmakologie   terapeutické užití   MeSH
• kousnutí a bodnutí mikrobiologie   MeSH
• lidé MeSH
• Pasteurella multocida patogenita   MeSH
• psi * zranění   MeSH
• senioři MeSH
• sepse * etiologie   mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• psi * zranění   MeSH
• senioři MeSH
• Publikační typ
• kazuistiky MeSH

Cíl: Zjistit výskyt potenciálně patogenních druhů babesií pro člověka v klíšťatech a v krvi psů a jelenů ve vybraných regionech České republiky. Prevalenci Babesia spp. v klíšťatech porovnat s výskytem jiných patogenů přenášených klíšťaty jako Borrelia spp., Anaplasma spp., Rickettsia spp. Materiál a metody: Vzorky klíšťat byly jednotlivě homogenizovány, ze vzorků klíšťat a krve živočichů provedena izolace DNA. Detekce Babesia spp. byla stanovena metodou PCR 18S rRNA genu a sekvenační analýzou PCR produktů určeny jednotlivé druhy babesií. Výsledky: V letech 2014–2016 byla analyzována klíšťata a krev psů a jelenů na různých místech České republiky. Ze souboru 675 klíšťat Ixodes ricinus dosahovala pozitivita na přítomnost Babesia spp. hodnot od 0,0 do 3,3 %. Sekvenační analýzou byly v klíšťatech identifikovány druhy Babesia venatorum, Babesia microti (patogenní druhy pro člověka) a druh Babesia capreoli. Prevalence Babesia spp. v klíšťatech byla v porovnání s výskytem jiných patogenů jako Borrelia burgdorferi s. l. (29,3 %), Anaplasma phagocytophilum (4,9 %) nižší a srovnatelná s Rickettsia spp. (1,6 %). U třetiny pozitivních klíšťat na babesie byla zjištěna koinfekce s Borrelia burgdorferi s. l. (B. venatorum – Borrelia garinii, Borrelia afzelii a B. microti – B. afzelii). Ze 109 vzorků krve psů bylo 3,7 % pozitivních na Babesia spp. s výskytem druhů Babesia gibsoni a Babesia vulpes. Z 50 vzorků krve jelenů z přírodního ekosystému dosahovala pozitivita 4,0 %. Identifikován byl druh Babesia divergens, nejvíce patogenní druh Babesia spp. pro člověka. Z 80 vzorků krve jelenů chovaných na farmách bylo pozitivních 5,0 % s výskytem druhu Babesia odocoilei. Nukleotidové sekvence babesií způsobujících humánní babesiózu byly zaslány do genové banky a přijaty pod čísly ON892053 (B. venatorum), ON892061 (B. microti), ON892067 (B. divergens). Závěr: Metodou PCR 18S rRNA genu a sekvenací amplikonů byly na území České republiky detekovány tři druhy babesií patogenních pro člověka: B. divergens, B. venatorum, B. microti. Výskyt těchto druhů babesií znamená potenciální riziko onemocnění babesiózou, zejména pro asplenické a imunokompromitované pacienty. Zjištěné koinfekce s Borrelia burgdorferi s. l. mohou být příčinou komplikovaného průběhu onemocnění.

Aim: To determine the occurrence of species of Babesia potentially pathogenic for humans in ticks and in the blood of dogs and deer in selected regions of the Czech Republic. To compare the prevalence of Babesia spp. in ticks with that of other tick-borne pathogens, such as Borrelia spp., Anaplasma spp., and Rickettsia spp. Material and Methods: Tick samples were individually homogenized. DNA was isolated from tick samples and animal blood. The detection of Babesia spp. was based on PCR of the 18S rRNA gene, and the identification to the species level was done by sequencing analysis of the PCR products. Results: In 2014–2016, ticks and blood of dogs and deer collected in various areas of the Czech Republic were analyzed. In a set of 675 Ixodes ricinus ticks, the positivity rate for Babesia spp. varied from 0.0 to 3.3 %. The species Babesia venatorum, Babesia microti (both pathogenic for humans), and Babesia capreoli were identified in ticks by sequencing analysis. The prevalence of Babesia spp. in ticks compared to that of other pathogens such as Borrelia burgdorferi s. l. (29.3 %) or Anaplasma phagocytophilum (4.9 %) was lower and comparable to that of Rickettsia spp. (1.6 %). Co-infection with Borrelia burgdorferi s.l (B. venatorum – Borrelia garinii, Borrelia afzelii, and B. microti – B. afzelii) was found in a third of Babesia spp. positive ticks. Out of 109 dog blood samples, 3.7 % were positive for Babesia spp., specifically Babesia gibsoni and Babesia vulpes. Of 50 blood samples of wild deer from the natural ecosystem, the positivity rate reached 4.0 %. The species Babesia divergens, a major human pathogen, was identified. Out of 80 blood samples from farmed deer, 5.0 % were positive for the species Babesia odocoilei. Nucleotide sequences of the agents causing human babesiosis were deposited in the gene bank under accession numbers ON892053 (B. venatorum), ON892061 (B. microti), and ON892067 (B. divergens). Conclusions: Using PCR of the 18S rRNA gene and amplicon sequencing, three species of Babesia causing human babesiosis were detected in the Czech Republic: B. divergens, B. venatorum, and B. microti. Babesia spp. pathogenic for humans pose a potential risk especially in asplenic and immunocompromised patients. The detected co-infections with Borrelia spp. can be the cause of a complicated course of the disease.

• MeSH
• Babesia mikrobiologie   MeSH
• babezióza * epidemiologie   krev   přenos   MeSH
• Borrelia burgdorferi MeSH
• diagnostické techniky molekulární metody   MeSH
• klíšťata * mikrobiologie   MeSH
• koinfekce diagnóza   přenos   MeSH
• krev mikrobiologie   MeSH
• lidé MeSH
• nemoci přenášené klíšťaty epidemiologie   přenos   prevence a kontrola   MeSH
• polymerázová řetězová reakce metody   MeSH
• psi * mikrobiologie   MeSH
• vysoká zvěř * krev   mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• psi * mikrobiologie   MeSH
• Geografické názvy
• Česká republika MeSH

Since the 1960s, more than 350,000 new chemicals have been introduced into the lives of humans and domestic animals. Many of them have become part of modern life and some are affecting nature as pollutants. Yet, our comprehension of their potential health risks for both humans and animals remains partial. The "epithelial barrier theory" suggests that genetic predisposition and exposure to diverse factors damaging the epithelial barriers contribute to the emergence of allergic and autoimmune conditions. Impaired epithelial barriers, microbial dysbiosis, and tissue inflammation have been observed in a high number of mucosal inflammatory, autoimmune and neuropsychiatric diseases, many of which showed increased prevalence in the last decades. Pets, especially cats and dogs, share living spaces with humans and are exposed to household cleaners, personal care products, air pollutants, and microplastics. The utilisation of cosmetic products and food additives for pets is on the rise, unfortunately, accompanied by less rigorous safety regulations than those governing human products. In this review, we explore the implications of disruptions in epithelial barriers on the well-being of companion animals, drawing comparisons with humans, and endeavour to elucidate the spectrum of diseases that afflict them. In addition, future research areas with the interconnectedness of human, animal, and environmental well-being are highlighted in line with the "One Health" concept.

• MeSH
• domácí zvířata * imunologie   MeSH
• epitel imunologie   MeSH
• kočky MeSH
• lidé MeSH
• psi MeSH
• vystavení vlivu životního prostředí škodlivé účinky   MeSH
• zvířata MeSH
• Check Tag
• kočky MeSH
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The cerebral biomarkers, neurofilament light chain (NfL), amyloid-β, tau, and neuron specific enolase (NSE) reflect a wide spectrum of neurological damage in the brain and spinal cord. With this study, we aimed to assess whether these biomarkers hold any potential diagnostic value for the three most common canine neurological diseases. Canines suffering from meningoencephalitis of unknown origin (MUO), brain tumors, and selected non-infectious myelopathies were included. For each diagnosis, we analyzed these biomarkers in the cerebrospinal fluid collected via cranial puncture from the cisterna magna. Elevated levels of CSF tau, NfL, and NSE were observed in MUO, with all three biomarkers being intercorrelated. Tau and NSE were increased while amyloid-β was decreased in dogs suffering from tumors. In contrast, no biomarker changes were observed in dogs with myelopathies. Covariates such as age, sex, or castration had minimal impact. CSF biomarkers may reflect molecular changes related to MUO and tumors, but not to non-infectious myelopathies. The combination of NfL, tau, and NSE may represent useful biomarkers for MUO as they reflect the same pathology and are not influenced by age.

• MeSH
• amyloidní beta-protein mozkomíšní mok   MeSH
• biologické markery * mozkomíšní mok   MeSH
• fosfopyruváthydratasa mozkomíšní mok   MeSH
• meningoencefalitida mozkomíšní mok   veterinární   diagnóza   MeSH
• nádory mozku mozkomíšní mok   veterinární   MeSH
• nemoci nervového systému mozkomíšní mok   veterinární   diagnóza   MeSH
• nemoci psů * mozkomíšní mok   diagnóza   MeSH
• neurofilamentové proteiny * mozkomíšní mok   MeSH
• proteiny tau * mozkomíšní mok   MeSH
• psi MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• psi MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

PIWI-interacting RNAs (piRNAs) play a crucial role in safeguarding genome integrity by silencing mobile genetic elements. From flies to humans, piRNAs originate from long single-stranded precursors encoded by genomic piRNA clusters. How piRNA clusters form to adapt to genomic invaders and evolve to maintain protection remain key outstanding questions. Here, we generate a roadmap of piRNA clusters across seven species that highlights both similarities and variations. In mammals, we identify transcriptional readthrough as a mechanism to generate piRNAs from transposon insertions (piCs) downstream of genes (DoG). Together with the well-known stress-dependent DoG transcripts, our findings suggest a molecular mechanism for the formation of piRNA clusters in response to retroviral invasion. Finally, we identify a class of dynamic piRNA clusters in humans, underscoring unique features of human germ cell biology. Our results advance the understanding of conserved principles and species-specific variations in piRNA biology and provide tools for future studies.

• MeSH
• druhová specificita MeSH
• lidé MeSH
• malá interferující RNA * metabolismus   genetika   MeSH
• myši MeSH
• Piwi-interagující RNA MeSH
• psi MeSH
• savci * genetika   MeSH
• transpozibilní elementy DNA genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH