Acute appendicitis as a manifestation of the immune reconstitution inflammatory syndrome is repeatedly discussed in the literature, but only a few cases of acute appendicitis associated with the initiation of cART have been published as yet. We describe a case of a young HIV-infected man who suffered from acute appendicitis early after the successful switch of a failing cART regimen.

• MeSH
• antiretrovirové látky škodlivé účinky   terapeutické užití   MeSH
• apendicitida etiologie   patologie   MeSH
• CD4-pozitivní T-lymfocyty imunologie   MeSH
• dospělí MeSH
• HIV infekce komplikace   farmakoterapie   MeSH
• imunorestituční zánětlivý syndrom chemicky indukované   komplikace   diagnóza   patologie   MeSH
• lidé MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Názvy látek
• antiretrovirové látky MeSH

In bone marrow transplantation (BMT), hematopoiesis-reconstituting cells are introduced following myeloablative treatment, which eradicates existing hematopoietic cells and disrupts stroma within the hematopoietic tissue. Both hematopoietic cells and stroma then undergo regeneration. Our study compares the outcomes of a second BMT administered to mice shortly after myeloablative treatment and the first BMT, with those of a second BMT administered to mice experiencing robust hematopoietic regeneration after the initial transplant. We evaluated the efficacy of the second BMT in terms of engraftment efficiency, types of generated blood cells, and longevity of function. Our findings show that regenerating hematopoiesis readily accommodates newly transplanted stem cells, including those endowed with a robust capacity for generating B and T cells. Importantly, our investigation uncovered a window for preferential engraftment of transplanted stem cells coinciding with the resumption of blood cell production. Repeated BMT could intensify hematopoiesis reconstitution and enable therapeutic administration of genetically modified autologous stem cells.

• Klíčová slova
• B cells, T cells, hematopoiesis, immune system, regeneration, stem cell, transplantation,
• MeSH
• hematopoetické kmenové buňky imunologie   MeSH
• hematopoéza * MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• regenerace MeSH
• rekonstituce imunitních funkcí MeSH
• transplantace kostní dřeně * metody   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Dehydrogenase/reductase SDR family member 7 (DHRS7, SDR34C1, retSDR4) is one of the many endoplasmic reticulum bound members of the SDR superfamily. Preliminary results indicate its potential significance in human metabolism. DHRS7 containing TEV-cleavable His10 and FLAG-tag expressed in the Sf9 cell line was solubilised, purified, and reconstituted into liposomes to enable the improved characterisation of this enzyme in the future. Igepal CA-630 was determined to be the best detergent for the solubilisation process. The solubilised DHRS7 was purified using affinity chromatography, and the purified enzyme was subjected to TEV cleavage of the affinity tags and then repurified using subtractive Ni-IMAC. The cleaved and uncleaved versions of DHRS7 were successfully reconstituted into liposomes. In addition, using tobacco specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) as the substrate, the cleaved liposomal DHRS7 was found to be inactive, whereas the pure and uncleaved liposomal DHRS7 were confirmed as enzymes, which reduce carbonyl group of the substrates.

• Klíčová slova
• Carbonyl reductase activity, DHRS7, Purification, Reconstitution, Short-chain dehydrogenases/reductases, Solubilisation,
• MeSH
• buněčná membrána MeSH
• lidé MeSH
• membránové proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• oxidoreduktasy chemie   genetika   izolace a purifikace   metabolismus   MeSH
• rekombinantní proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• Sf9 buňky MeSH
• Spodoptera MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• membránové proteiny MeSH
• oxidoreduktasy MeSH
• rekombinantní proteiny MeSH

Intracellular trafficking of organelles driven by molecular motors underlies essential cellular processes. Mitochondria, the powerhouses of the cell, are one of the major cargoes of molecular motors. Efficient distribution of mitochondria ensures cellular fitness while defects in this process contribute to severe pathologies, such as neurodegenerative diseases. Reconstitution of the mitochondrial microtubule-based transport in vitro in a bottom-up approach provides a powerful tool to investigate the mitochondrial trafficking machinery in a controlled environment in the absence of complex intracellular interactions. In this chapter, we describe the procedures for achieving such reconstitution of mitochondrial transport.

• Klíčová slova
• Adaptor proteins, Interference reflection microscopy, Kinesin-1, Mitochondria, Molecular motors, Motility assay, TIRF microscopy, TRAK,
• MeSH
• biologický transport MeSH
• kineziny * MeSH
• mikrotubuly * metabolismus   MeSH
• mitochondrie metabolismus   MeSH
• organely MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kineziny * MeSH

Reconstitution of novel mitochondrial uncoupling proteins, human UCP2 and UCP3, expressed in yeast, was performed to characterize fatty acid (FA)-induced H+ efflux in the resulted proteoliposomes. We now demonstrate for the first time that representatives of physiologically abundant long chain FAs, saturated or unsaturated, activate H+ translocation in UCP2- and UCP3-proteoliposomes. Efficiency of lauric, palmitic or linoleic acid was roughly the same, but oleic acid induced faster H+ uniport. We have confirmed that ATP and GTP inhibit such FA-induced H+ uniport mediated by UCP2 and UCP3. Coenzyme Q10 did not further significantly activate the observed H+ efflux. In conclusion, careful instant reconstitution yields intact functional recombinant proteins, UCP2 and UCP3, the activity of which is comparable with UCP1.

• MeSH
• iontové kanály MeSH
• kvasinky metabolismus   MeSH
• lidé MeSH
• mastné kyseliny metabolismus   MeSH
• membránové transportní proteiny * MeSH
• mitochondriální proteiny * MeSH
• mitochondrie metabolismus   MeSH
• proteiny metabolismus   MeSH
• transportní proteiny metabolismus   MeSH
• uncoupling protein 2 MeSH
• uncoupling protein 3 MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• iontové kanály MeSH
• mastné kyseliny MeSH
• membránové transportní proteiny * MeSH
• mitochondriální proteiny * MeSH
• proteiny MeSH
• transportní proteiny MeSH
• UCP2 protein, human MeSH Prohlížeč
• UCP3 protein, human MeSH Prohlížeč
• uncoupling protein 2 MeSH
• uncoupling protein 3 MeSH

Carotenoids are crucial for photosynthesis, playing key roles in light harvesting and photoprotection. In this study, spheroidene and bacteriochlorophyll a (Bchl a) were reconstituted into the chromatophores of the carotenoidless mutant Rhodobacter sphaeroides R26.1, resulting in the preparation of high-quality LH2 complexes. Global and target analyses of transient absorption data revealed that incorporating B800 Bchl a significantly enhances excitation energy transfer (EET) efficiency from carotenoids to Bchl a. EET predominantly occurs from the carotenoid S2 state, with additional pathways from the S1 state observed in native LH2. Unique relaxation dynamics were identified, including the generation of the carotenoid S* state in reconstituted LH2 with both spheroidene and B800 Bchl a and the formation of the carotenoid T1 state in reconstituted LH2. These findings underscore the critical influence of pigment composition and spatial organization on energy transfer mechanisms. They provide valuable insights into the molecular interplay that governs excitation energy transfer in photosynthetic light-harvesting systems.

• Klíčová slova
• B800 bacteriochlorophyll a, carotenoid, light-harvesting, photoprotection, purple photosynthetic bacteria, reconstitution,
• MeSH
• bakteriální proteiny metabolismus   genetika   chemie   MeSH
• bakteriochlorofyl A * metabolismus   chemie   MeSH
• fotosyntéza MeSH
• karotenoidy * metabolismus   chemie   MeSH
• přenos energie * MeSH
• Rhodobacter sphaeroides * metabolismus   genetika   MeSH
• světlosběrné proteinové komplexy * metabolismus   chemie   genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• bakteriální proteiny MeSH
• bakteriochlorofyl A * MeSH
• karotenoidy * MeSH
• světlosběrné proteinové komplexy * MeSH

αβT-cell-depleted allogeneic hematopoietic cell transplantation holds promise for the safe and accessible therapy of both malignant and non-malignant blood disorders. Here we employed molecular barcoding normalized T-cell receptor (TCR) profiling to quantitatively track T-cell immune reconstitution after TCRαβ-/CD19-depleted transplantation in children. We demonstrate that seemingly early reconstitution of αβT-cell counts 2 months after transplantation is based on only several hundred rapidly expanded clones originating from non-depleted graft cells. In further months, frequency of these hyperexpanded clones declines, and after 1 year the observed T-cell counts and TCRβ diversity are mostly provided by the newly produced T cells. We also demonstrate that high TCRβ diversity at day 60 observed for some of the patients is determined by recipient T cells and intrathymic progenitors that survived conditioning regimen. Our results indicate that further efforts on optimization of TCRαβ-/CD19-depleted transplantation protocols should be directed toward providing more efficient T-cell defense in the first months after transplantation.

• MeSH
• antigeny CD19 * MeSH
• časové faktory MeSH
• dítě MeSH
• kojenec MeSH
• krevní nemoci terapie   MeSH
• lidé MeSH
• lymfocytární deplece metody   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• předškolní dítě MeSH
• přežívání štěpu * MeSH
• receptory antigenů T-buněk alfa-beta * MeSH
• T-lymfocyty imunologie   MeSH
• transplantace hematopoetických kmenových buněk metody   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• předškolní dítě MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD19 * MeSH
• receptory antigenů T-buněk alfa-beta * MeSH

Severe combined immunodeficient (SCID) mice develop lethal infections, resembling opportunistic microsporidiosis of immunocompromised patients, after intraperitoneal (i.p.) inoculations of spores of Encephalitozoon cuniculi. In the present study, SCID mice reconstituted i.p. with 5 x 10(7) spleen cells from naive adult BALB/c mice 14 days prior to the i.p. injection of 10(7) spores were completely resistant to the infection, whereas control infected SCID mice developed clinical disease and died within 17 days post infection (DPI). In another experiment, SCID mice infected i.p. with 10(7) spores of E. cuniculi and after that (on DPI 7) injected i.p. with 5 x 10(7) spleen lymphocytes isolated from immune adult BALB/c mice were partially protected against the parasite (40% of the reconstituted mice survived). In both experiments, high levels of parasite-specific serum antibodies (mostly of the IgG-isotype) were detected in the infected immunocompetent BALB/c mice, whereas virtually no antibodies were found in the infected SCID mice. However, SCID mice reconstituted with either naive spleen cells or immune lymphocytes revealed humoral immune responses comparable with those of immunocompetent mice.

• MeSH
• Encephalitozoon patogenita   MeSH
• encephalitozoonóza imunologie   terapie   MeSH
• imunoterapie adoptivní * MeSH
• myši inbrední BALB C MeSH
• myši SCID MeSH
• myši MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Research in the area of security for Wireless Sensor Networks over the past two decades has yielded many interesting findings. We focus on the topic of (re-)securing link keys between sensor nodes through so-called secrecy amplification (SA) protocols. Crowdsourcing is at the very heart of these SA protocols. Not only do SA protocols work wonders even for low-level constrained nodes with no tamper resistance, they exhibit astonishing performance in networks under significant attacker control. Our work shows that even when 50% of all network links are compromised, SA protocols can re-secure over 90% of the link keys through an intriguingly simple crowdsourcing mechanism. These protocols allow us to re-take control without any broadly coordinated cooperation, without knowledge of the compromised links, with only very limited knowledge of each particular network node and independently of decisions made by other nodes. Our article first outlines the principles of and presents existing approaches to SA, introducing most of the important related concepts, then presents novel conclusive results for a realistic attacker model parametrised by attacker behaviour and capabilities. We undertook this work using two very different simulators, and we present here the results of analyses and detailed comparisons that have not previously been available. Finally, we report the first real, non-simulated network test results for the most attractive SA protocol, our implementations of which are available as open-source code for two platforms: Arduino and TinyOS. This work demonstrates the practical usability (and the attractive performance) of SA, serving as a ripe technology enabler for (among others) networks with many potentially compromised low-level devices.

• Klíčová slova
• ad hoc networks, crowdsourcing, cryptographic protocol, secrecy amplification (SA), wireless sensor network (WSN),
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Galectin-7 (Gal-7) is negatively regulated in cervical cancer, and appears to be a link between the apoptotic response triggered by cancer and the anti-tumoral activity of the immune system. Our understanding of how cervical cancer cells and their molecular networks adapt in response to the expression of Gal-7 remains limited. METHODS: Meta-analysis of Gal-7 expression was conducted in three cervical cancer cohort studies and TCGA. In silico prediction and bisulfite sequencing were performed to inquire epigenetic alterations. To study the effect of Gal-7 on cervical cancer, we ectopically re-expressed it in the HeLa and SiHa cervical cancer cell lines, and analyzed their transcriptome and SILAC-based proteome. We also examined the tumor and microenvironment host cell transcriptomes after xenotransplantation into immunocompromised mice. Differences between samples were assessed with the Kruskall-Wallis, Dunn's Multiple Comparison and T tests. Kaplan-Meier and log-rank tests were used to determine overall survival. RESULTS: Gal-7 was constantly downregulated in our meta-analysis (p < 0.0001). Tumors with combined high Gal-7 and low galectin-1 expression (p = 0.0001) presented significantly better prognoses (p = 0.005). In silico and bisulfite sequencing assays showed de novo methylation in the Gal-7 promoter and first intron. Cells re-expressing Gal-7 showed a high apoptosis ratio (p < 0.05) and their xenografts displayed strong growth retardation (p < 0.001). Multiple gene modules and transcriptional regulators were modulated in response to Gal-7 reconstitution, both in cervical cancer cells and their microenvironments (FDR < 0.05 %). Most of these genes and modules were associated with tissue morphogenesis, metabolism, transport, chemokine activity, and immune response. These functional modules could exert the same effects in vitro and in vivo, even despite different compositions between HeLa and SiHa samples. CONCLUSIONS: Gal-7 re-expression affects the regulation of molecular networks in cervical cancer that are involved in diverse cancer hallmarks, such as metabolism, growth control, invasion and evasion of apoptosis. The effect of Gal-7 extends to the microenvironment, where networks involved in its configuration and in immune surveillance are particularly affected.

• Klíčová slova
• Cervical cancer, Differential network analysis, Galectin-7, Microenvironment crosstalk,
• MeSH
• galektiny metabolismus   MeSH
• lidé MeSH
• nádorové mikroprostředí fyziologie   MeSH
• nádory děložního čípku metabolismus   patologie   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• metaanalýza MeSH
• práce podpořená grantem MeSH
• Názvy látek
• galektiny MeSH
• LGALS7 protein, human MeSH Prohlížeč