BACKGROUND: The treatment of non-small cell lung cancer (NSCLC) patients is correlated with the efficacy of immune checkpoint blockade therapy (ICB) targeting programmed cell death ligand 1 (PD-L1) or its cognate receptor (PD-1) on cancer cells or infiltrating immune cells. Analysis of PD-L1/PD-1 expression in tumor tissue represents a crucial step before PD-L1/PD-1 blocker usage. METHODS: We used directed evolution of protein variants derived from a 13 kDa Myomedin loop-type combinatorial library with 12 randomized amino acid residues to select high-affinity binders of human PD-L1 (hPD-L1). After the ribosome display, individual clones were screened by ELISA. Detailed analysis of binding affinity and kinetics was performed using LigandTracer. The specificity of Myomedins was assessed using fluorescent microscopy on HEK293T-transfected cells and cultured cancer cells in vitro, formalin-fixed paraffin-embedded (FFPE) sections of human tonsils, and FFPE tumor samples of NSCLC patients. RESULTS: Seven identified PD-L1 binders, called MLE, showed positive staining for hPD-L1 on transfected HEK293T cells and cultured MCF-7 cells. MLE031, MLE105, MLE249, and MLE309 exhibited high affinity to both human and mouse PD-L1-transfected HEK293T cells measured with LigandTracer. The diagnostic potential of MLE variants was tested on human tonsillitis tissue and compared with diagnostic anti-PD-L1 antibody DAKO 28-8 and PD-L1 IHC 22C3 pharmDx antibody. MLE249 and MLE309 exhibited an excellent overlap with diagnostic DAKO 28-8 (Pearson ́s coefficient (r) = 0.836 and 0.731, respectively) on human tonsils on which MLE309 exhibited also excellent overlap with diagnostic 22C3 antibody (r = 0.876). Using three NSCLC tissues, MLE249 staining overlaps with 28-8 antibody (r = 0.455-0.883), and MLE309 exhibited overlap with 22C3 antibody (r = 0.534-0.619). Three MLE proteins fused with Fc fragments of rabbit IgG, MLE249-rFc, MLE309-rFc and MLE031-rFc, exhibited very good overlap with anti-PD-L1 antibody 28-8 on tonsil tissue (r = 0.691, 0.610, and 0.667, respectively). Finally, MLE249-rFc, MLE309-rFc and MLE031-rFc exhibited higher sensitivity in comparison to IHC 22C3 antibody using routine immunohistochemistry staining system Ventana, which is one of gold standards for PD-L1 diagnosis. CONCLUSIONS: We demonstrated the development of MLE Myomedins specifically recognizing hPD-L1 that may serve as a refinement tool for clinical PD-L1 detection.

• MeSH
• antigeny CD274 * metabolismus   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory plic * diagnóza   metabolismus   patologie   MeSH
• nemalobuněčný karcinom plic * diagnóza   metabolismus   patologie   MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Intravenous immunoglobulins (IVIG) are commonly used in peri-transplant desensitization, but evidence supporting their efficacy is limited. We conducted a prospective, randomized single-center, open-label, Phase IIIb non-inferiority clinical pilot trial to compare the efficacy of IVIG (administered at a dose of 3 × 0.5 g/kg) versus no IVIG, in conjunction with rabbit anti-thymocyte globulin (5-7 mg/kg) induction, in pre-sensitized patients with donor-specific antibodies who had negative pre-transplantation Flow- and CDC-crossmatches, between July 2020 and November 2022. The primary endpoint was the rate of efficacy failure, defined as biopsy-proven rejection within 12-month post-transplant. Secondary endpoints included the incidence of rejection at protocol biopsies, evaluated by histology and biopsy-based transcripts diagnostics. Of the screened patients, 53 (72.6%) were excluded due to crossmatch positivity. Ten patients were randomized to the IVIG+, and 7 to the IVIG-arm. The trial was prematurely terminated due to futility at interim analysis. In the IVIG-arm, 3 patients (43%) experienced the primary endpoint compared to none in the IVIG+ arm (p = 0.026). MMDx identified one molecular ABMR in the IVIG+ and 2 in the IVIG-arm in 12-month protocol biopsies. There was one graft loss in the IVIG-arm. The results of this pilot study, although not definitive, do not support the use of IVIG-sparing regimens in HLA-incompatible kidney transplantation (NCT04302805). This study is registered on ClinicalTrials.gov under the identifier NCT04302805.

• MeSH
• antilymfocytární sérum * aplikace a dávkování   MeSH
• desenzibilizace imunologická * metody   MeSH
• dospělí MeSH
• intravenózní imunoglobuliny * aplikace a dávkování   MeSH
• lidé středního věku MeSH
• lidé MeSH
• pilotní projekty MeSH
• prospektivní studie MeSH
• rejekce štěpu * prevence a kontrola   imunologie   MeSH
• transplantace ledvin * škodlivé účinky   MeSH
• výsledek terapie MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky, fáze III MeSH
• randomizované kontrolované studie MeSH

Článek se zabývá stručným popisem zdravotních problémů, na něž má vliv výživa. Nejčastějšími zdravotními problémy králíků se vztahem k výživě jsou onemocnění gastrointestinálního traktu (trichobezoáry, enteritidy, enterotoxemie), obezita a syndrom onemocnění dentice (a s tím související problémy). Na tyto problémy pak navazují další problémy a změny chování králíků. V závěru je stručně uvedeno správné složení krmné dávky králíků v zájmovém chovu.

The article deals with a description of the health problems affected by nutrition. The most common nutrition-related health problems in rabbits are gastrointestinal diseases (trichobezoars, enteritis, enterotoxaemia), obesity and dentition syndrome (and related problems). These problems are then followed by other problems and behavioural changes in rabbits. In conclusion, the correct composition of rabbit diets for pet rabbits is briefly outlined.

• MeSH
• chov zvířat MeSH
• enteropatogenní Escherichia coli MeSH
• exsudativní enteropatie etiologie   veterinární   MeSH
• fyziologie výživy zvířat MeSH
• králíci MeSH
• nemoci trávicího systému * etiologie   veterinární   MeSH
• nemoci zvířat * MeSH
• pohoda zvířat MeSH
• rostlinná strava MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• přehledy MeSH

OBJECTIVES: Treponema pallidum subsp. pallidum (T. pallidum) is the etiological agent of syphilis, a sexually transmitted disease of global public health importance. The objective of this study was to introduce a novel in vitro protocol for isolation of T. pallidum directly from patients' clinical samples, eliminating the need for rabbit propagation. METHODS: Four oral and five genital swabs were collected from nine epidemiologically unrelated patients at two hospitals in Brno, Czech Republic. Swabs were submerged in TpCM-2 medium for transport. Samples were then placed on a 0.4 μm filters and incubated for 2.5 hours. During this period, spiral T. pallidum cells passed through the filter pores to the well containing TpCM-2 medium and rabbit feeder cells (Sf1Ep). Stable T. pallidum cultures (containing >1 × 107 treponemes) were achieved by subculturing every 7 days into fresh well. RESULTS: A successful protocol for in vitro isolation of T. pallidum was established. From the nine clinical specimens processed, six T. pallidum cultures (MU1-MU6) were derived after 14 to 112 days of cultivation. Five of these strains (MU1-MU5) belonged to SS14-like cluster and shared the same allelic profile 1.3.1. The remaining strain (MU6) was identified as a Nichols-like strain with an allelic profile 9.16.3. DISCUSSION: The introduced in vitro protocol enables isolation of T. pallidum from clinical material, including frozen samples, without the need for experimental rabbits. This method facilitates the isolation of contemporary, clinically relevant treponemal strains.

• MeSH
• bakteriologické techniky * metody   MeSH
• králíci MeSH
• lidé MeSH
• pohlavní orgány mikrobiologie   MeSH
• syfilis * mikrobiologie   diagnóza   MeSH
• Treponema pallidum * izolace a purifikace   genetika   klasifikace   MeSH
• ústa mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Free radical polymerization technique was used to formulate Poloxamer-188 based hydrogels for controlled delivery. A total of seven formulations were formulated with varying concentrations of polymer, monomer ad cross linker. In order to assess the structural properties of the formulated hydrogels, Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetric analysis (TGA), Differential Scanning Calorimetry (DSC), Scanning electron microscopy (SEM), and X-ray diffraction (XRD) were carried out. To assess the effect of pH on the release of the drug from the polymeric system, drug release studies were carried in pH 1.2 and 7.4 and it was found that release of the drug was significant in pH 7.4 as compared to that of pH 1.2 which confirmed the pH responsiveness of the system. Different kinetic models were also applied to the drug release to evaluate the mechanism of the drug release from the system. To determine the safety and biocompatibility of the system, toxicity study was also carried out for which healthy rabbits were selected and formulated hydrogels were orally administered to the rabbits. The results obtained suggested that the formulated poloxamer-188 hydrogels are biocompatible with biological system and have the potential to serve as controlled drug delivery vehicles.

• MeSH
• akrylové pryskyřice * chemie   MeSH
• diferenciální skenovací kalorimetrie MeSH
• difrakce rentgenového záření MeSH
• hydrogely * chemie   MeSH
• koncentrace vodíkových iontů MeSH
• králíci MeSH
• lékové transportní systémy MeSH
• léky s prodlouženým účinkem chemie   farmakokinetika   MeSH
• mikroskopie elektronová rastrovací MeSH
• nosiče léků chemie   MeSH
• poloxamer * chemie   MeSH
• spektroskopie infračervená s Fourierovou transformací MeSH
• termogravimetrie MeSH
• timolol * aplikace a dávkování   farmakokinetika   chemie   MeSH
• uvolňování léčiv MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Virus hepatitidy E (HEV) je celosvětovým problémem a druhou nejčastější příčinou virové hepatitidy. Existuje osm genotypů HEV, z nichž čtyři způsobují onemocnění u lidí. Genotypy 1 a 2 se přenášejí kontaminovanou vodou, zatímco genotypy 3 a 4 jsou zoonotické a přenášejí se konzumací nedostatečně tepelně upraveného masa, zejména vepřového, zvěřiny a králičího. V České republice je endemický genotyp 3 s rezervoárem u prasat, spárkaté zvěře a králíků. Infekce HEV se projevuje podobně jako jiné typy hepatitid příznaky, jako je únava, horečka, nevolnost, žloutenka a tmavá moč. U imunokompetentních pacientů infekce obvykle odezní sama, ale u imunokompromitovaných a imusuprimovaných může přejít do chronického stavu vedoucího až k cirhóze a selhání jater. Preexistující onemocnění jater hraje v kontextu HEV významnou roli, jelikož u pacientů s chronickým onemocněním jater infikovaných HEV existuje významně vyšší riziko selhání jater, které může vést až k úmrtí. Diagnóza se provádí detekcí protilátek IgM a IgG proti HEV a HEV RNA v krvi a stolici. Doporučené diagnostické postupy se liší v závislosti na imunitním stavu pacienta a epidemiologickém prostředí. Léčba obvykle není nutná, ale v závažných případech se používá ribavirin a interferon a. Prevence zahrnuje zlepšení hygieny, dostatečnou tepelnou úpravu masa a v rozvojových zemích zvážení očkování. V Číně je dostupná vakcína Hecolin® účinná proti genotypům 1, 2 a 4, ale v Evropské unii zatím není registrována.

The hepatitis E virus (HEV) represents a significant global health concern, accounting for the second-highest incidence of viral hepatitis. Eight genotypes of HEV have been identified, four of which are associated with disease in humans. Genotypes 1 and 2 are transmitted via contaminated water, whereas genotypes 3 and 4 are zoonotic and transmitted through the consumption of undercooked meat, particularly pork, venison, and rabbit. Genotype 3 is endemic in the Czech Republic, with reservoirs in pigs, wild game, and rabbits. A hepatitis E virus (HEV) infection presents with symptoms similar to those observed in other types of hepatitis, including fatigue, fever, malaise, jaundice, and dark urine. In immunocompetent patients, the infection typically resolves spontaneously. However, in immunocompromised patients, it can progress to a chronic state, leading to cirrhosis and liver failure. The presence of preexisting liver disease is a significant factor in the context of HEV, as patients with chronic liver disease infected with HEV are at an elevated risk of liver failure, which can potentially result in death. A diagnosis is made by detecting immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies to the hepatitis E virus (HEV) and HEV ribonucleic acid (RNA) in blood and stool samples. The recommended diagnostic procedures are contingent upon the patient‘s immune status and epidemiological context. In most cases, treatment is not necessary; however, ribavirin and interferon a are employed in severe instances. Prevention strategies encompass enhanced hygiene practices, thorough cooking of meat, and in developing countries, the consideration of vaccination. The vaccine Hecolin® is available in China and is efficacious against genotypes 1, 2, and 4; however, it is not yet registered in the European Union.

• MeSH
• genotyp MeSH
• hepatitida E * diagnóza   farmakoterapie   přenos   prevence a kontrola   MeSH
• lidé MeSH
• virus hepatitidy E patogenita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH
• Geografické názvy
• Česká republika MeSH

In this study, we investigated the mechanism underlying electrocardiogram (ECG) alterations in a rabbit model of acute pulmonary thromboembolism (PTE). Twelve healthy adult New Zealand white rabbits were used, with eight in the experimental group (PTE group) and four in the control group. After developing the rabbit model of acute PTE, ECG and coronary angiography were performed. HE staining was conducted on the right and left ventricular tissues, and polymerase chain reaction (PCR) was used to determine brain natriuretic peptide (BNP), tumor necrosis factor-alpha (TNF-?), and Troponin I (TNI) mRNA expression in the myocardium. There were considerable changes in the ST segment of the ECG in the PTE group. Coronary angiography revealed the absence of spasm, stenosis, and occlusion. In the plasma of the PTE group, the levels of D-dimer, BNP, TNF-?, and TNI were significantly elevated, and these changes were statistically significant (P<0.05). PCR analysis of ventricular myocardial tissue indicated significantly higher levels of BNP, TNF-?, and TNI mRNA in the PTE group than in the control group. These differences were statistically significant (P<0.05). The ST-T variations on the ECG of rabbits with acute PTE correlate strongly with the temporary changes in right heart volume caused by acute PTE. Keywords: Animal model of pulmonary embolism, B-type natriuretic peptide, Electrocardiogram, Pulmonary thromboembolism, Troponin I, Tumor necrosis factor-alpha.

• MeSH
• akutní nemoc MeSH
• elektrokardiografie * MeSH
• králíci MeSH
• modely nemocí na zvířatech * MeSH
• natriuretický peptid typu B krev   MeSH
• plicní embolie * patofyziologie   krev   MeSH
• TNF-alfa krev   metabolismus   genetika   MeSH
• troponin I krev   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The increase in syphilis rates worldwide necessitates development of a vaccine with global efficacy. We aimed to explore Treponema pallidum subspecies pallidum (TPA) molecular epidemiology essential for vaccine research by analysing clinical data and specimens from early syphilis patients using whole-genome sequencing (WGS) and publicly available WGS data. METHODS: In this multicentre, cross-sectional, molecular epidemiology study, we enrolled patients with primary, secondary, or early latent syphilis from clinics in China, Colombia, Malawi, and the USA between Nov 28, 2019, and May 27, 2022. Participants aged 18 years or older with laboratory confirmation of syphilis by direct detection methods or serological testing, or both, were included. Patients were excluded from enrolment if they were unwilling or unable to give informed consent, did not understand the study purpose or nature of their participation, or received antibiotics active against syphilis in the past 30 days. TPA detection and WGS were conducted on lesion swabs, skin biopsies, skin scrapings, whole blood, or rabbit-passaged isolates. We compared our WGS data to publicly available genomes and analysed TPA populations to identify mutations associated with lineage and geography. FINDINGS: We screened 2802 patients and enrolled 233 participants, of whom 77 (33%) had primary syphilis, 154 (66%) had secondary syphilis, and two (1%) had early latent syphilis. The median age of participants was 28 years (IQR 22-35); 154 (66%) participants were cisgender men, 77 (33%) were cisgender women, and two (1%) were transgender women. Of the cisgender men, 66 (43%) identified as gay, bisexual, or other sexuality. Among all participants, 56 (24%) had HIV co-infection. WGS data from 113 participants showed a predominance of SS14-lineage strains with geographical clustering. Phylogenomic analyses confirmed that Nichols-lineage strains were more genetically diverse than SS14-lineage strains and clustered into more distinct subclades. Differences in single nucleotide variants (SNVs) were evident by TPA lineage and geography. Mapping of highly differentiated SNVs to three-dimensional protein models showed population-specific substitutions, some in outer membrane proteins (OMPs) of interest. INTERPRETATION: Our study substantiates the global diversity of TPA strains. Additional analyses to explore TPA OMP variability within strains is vital for vaccine development and understanding syphilis pathogenesis on a population level. FUNDING: US National Institutes of Health National Institute for Allergy and Infectious Disease, the Bill & Melinda Gates Foundation, Connecticut Children's, and the Czech Republic National Institute of Virology and Bacteriology.

• MeSH
• bakteriální vakcíny imunologie   aplikace a dávkování   MeSH
• dospělí MeSH
• fylogeneze MeSH
• genetická variace genetika   MeSH
• genom bakteriální MeSH
• genomika MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• molekulární epidemiologie * MeSH
• průřezové studie MeSH
• sekvenování celého genomu * MeSH
• syfilis * epidemiologie   mikrobiologie   MeSH
• Treponema pallidum * genetika   imunologie   MeSH
• Treponema MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• Geografické názvy
• Spojené státy americké MeSH

The treponemes infecting lagomorphs include Treponema paraluisleporidarum ecovar Cuniculus (TPeC) and ecovar Lepus (TPeL), infecting rabbits and hares, respectively. In this study, we described the first complete genome sequence of TPeL, isolate V3603-13, from an infected mountain hare (Lepus timidus) in Sweden. In addition, we determined 99.0% of the genome sequence of isolate V246-08 (also from an infected mountain hare, Sweden) and 31.7% of the genome sequence of isolate Z27 A77/78 (from a European hare, Lepus europeaus, The Netherlands). The TPeL V3603-13 genome had considerable gene synteny with the TPeC Cuniculi A genome and with the human pathogen T. pallidum, which causes syphilis (ssp. pallidum, TPA), yaws (ssp. pertenue, TPE) and endemic syphilis (ssp. endemicum, TEN). Compared to the TPeC Cuniculi A genome, TPeL V3603-13 contained four insertions and 11 deletions longer than three nucleotides (ranging between 6 and2,932 nts). In addition, there were 25 additional indels, from one to three nucleotides long, altogether spanning 36 nts. The number of single nucleotide variants (SNVs) between TPeC Cuniculi A and TPeL V3603-13 were represented by 309 nucleotide differences. Major proteome coding differences between TPeL and TPeC were found in the tpr gene family, and (predicted) genes coding for outer membrane proteins, suggesting that these components are essential for host adaptation in lagomorph syphilis. The phylogeny revealed that the TPeL sample from the European brown hare was more distantly related to TPeC Cuniculi A than V3603-13 and V246-08.

• MeSH
• fylogeneze * MeSH
• genom bakteriální MeSH
• králíci MeSH
• syfilis * mikrobiologie   MeSH
• Treponema * genetika   izolace a purifikace   MeSH
• zajíci * mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Tento článek pojednává o chirurgickém řešení hydronefrózy (hydronefros) u zakrslého králíka. Jedná se o polymorbidního pacienta s kon kurentním mediastinálním thymomem a otitis media. Na případu našeho pacienta je popsaný laterální chirurgický přístup k nefrektomii, pooperační péče a následný vývoj zdravotního stavu.

This article describes the case of surgically treatment for hydronephrosis in a pet rabbit. Our rabbit is a polymorbidic patient with con current mediastinal thymoma and otitis media. In the case of our patient, the lateral surgical treatment for nephrectomy, postoperative care and subsequent development of overall status are described.

• MeSH
• hydronefróza * diagnostické zobrazování   diagnóza   patologie   MeSH
• králíci MeSH
• nefrektomie metody   MeSH
• radioisotopová renografie MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• kazuistiky MeSH