Here we described a new trypanosomatid species, Phytomonas lipae, parasitizing the dock bug Coreus marginatus based on axenic culture and in vivo material. Using light and electron microscopy we characterized the development of this flagellate in the intestine, hemolymph and salivary glands of its insect host. The intestinal promastigotes of Phytomonas lipae do not divide and occur only in the anterior part of the midgut. From there they pass into hemolymph, increasing in size, and then to salivary glands, where they actively proliferate without attachment to the host's epithelium and form infective endomastigotes. We conducted molecular phylogenetic analyses based on 18s rRNA, gGAPDH and HSP83 gene sequences, of which the third marker performed the best in terms of resolving phylogenetic relationships within the genus Phytomonas. Our inference demonstrated rather early origin of the lineage comprising the new species, right after that of P. oxycareni, which represents the earliest known branch within the Phytomonas clade. This allowed us to compare the development of P. lipae and three other Phytomonas spp. in their insect hosts and reconstruct the vectorial part of the life cycle of their common ancestor.

• MeSH
• fylogeneze MeSH
• Heteroptera parazitologie   MeSH
• Kinetoplastida MeSH
• pravděpodobnostní funkce MeSH
• proteiny tepelného šoku genetika   MeSH
• protozoální proteiny genetika   MeSH
• RNA ribozomální 18S genetika   MeSH
• slinné žlázy parazitologie   MeSH
• stadia vývoje * MeSH
• střeva parazitologie   MeSH
• Trypanosomatina klasifikace   genetika   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Clostridium beijerinckii NRRL B-598 is a sporulating, butanol and hydrogen producing strain that utilizes carbohydrates by the acetone-butanol-ethanol (ABE) fermentative pathway. The pathway consists of two metabolic phases, acidogenesis and solventogenesis, from which the latter one can be coupled with sporulation. Thorough transcriptomic profiling during a complete life cycle and both metabolic phases completed with flow cytometry, microscopy and a metabolites analysis helped to find out key genes involved in particular cellular events. The description of genes/operons that are closely involved in metabolism or the cell cycle is a necessary condition for metabolic engineering of the strain and will be valuable for all C. beijerinckii strains and other Clostridial species. The study focused on glucose transport and catabolism, hydrogen formation, metabolic stress response, binary fission, motility/chemotaxis and sporulation, which resulted in the composition of the unique image reflecting clostridial population changes. Surprisingly, the main change in expression of individual genes was coupled with the sporulation start and not with the transition from acidogenic to solventogenic metabolism. As expected, solvents formation started at pH decrease and the accumulation of butyric and acetic acids in the cultivation medium.

• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• Clostridium beijerinckii cytologie   genetika   MeSH
• fermentace genetika   MeSH
• fyziologický stres * genetika   MeSH
• glukosa metabolismus   MeSH
• kyseliny metabolismus   MeSH
• mastné kyseliny metabolismus   MeSH
• proteiny tepelného šoku genetika   metabolismus   MeSH
• regulace genové exprese u bakterií * MeSH
• rozpouštědla metabolismus   MeSH
• spory bakteriální metabolismus   MeSH
• transkriptom genetika   MeSH
• vodík metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The human iPSC cell line, CARS-FiPS4F1 (ESi064-A), derived from dermal fibroblast from the apparently healthy carrier of the mutation of the gene SACSIN, was generated by non-integrative reprogramming technology using OCT3/4, SOX2, CMYC and KLF4 reprogramming factors. The pluripotency was assessed by immunocytochemistry and RT-PCR. This iPSC line can be used as control for Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) disease.

• MeSH
• dospělí MeSH
• indukované pluripotentní kmenové buňky metabolismus   MeSH
• lidé MeSH
• mutace MeSH
• proteiny tepelného šoku genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH

Plum pox virus (PPV, family Potyviridae) is one of the most important viral pathogens of Prunus spp. causing considerable damage to stone-fruit industry worldwide. Among the PPV strains identified so far, only PPV-C, PPV-CR, and PPV-CV are able to infect cherries under natural conditions. Herein, we evaluated the pathogenic potential of two viral isolates in herbaceous host Nicotiana benthamiana. Significantly higher accumulation of PPV capsid protein in tobacco leaves infected with PPV-CR (RU-30sc isolate) was detected in contrast to PPV-C (BY-101 isolate). This result correlated well with the symptoms observed in the infected plants. To further explore the host response upon viral infection at the molecular level, a comprehensive proteomic profiling was performed. Using reverse-phase ultra-high-performance liquid chromatography followed by label-free mass spectrometry quantification, we identified 38 unique plant proteins as significantly altered due to the infection. Notably, the abundances of photosynthesis-related proteins, mainly from the Calvin-Benson cycle, were found more aggressively affected in plants infected with PPV-CR isolate than those of PPV-C. This observation was accompanied by a significant reduction in the amount of photosynthetic pigments extracted from the leaves of PPV-CR infected plants. Shifts in the abundance of proteins that are involved in stimulation of photosynthetic capacity, modification of amino acid, and carbohydrate metabolism may affect plant growth and initiate energy formation via gluconeogenesis in PPV infected N. benthamiana. Furthermore, we suggest that the higher accumulation of H2O2 in PPV-CR infected leaves plays a crucial role in plant defense and development by activating the glutathione synthesis.

• MeSH
• chlorofyl biosyntéza   MeSH
• chromatografie s reverzní fází MeSH
• energetický metabolismus genetika   MeSH
• fotosyntéza genetika   MeSH
• genotyp MeSH
• glutathion biosyntéza   MeSH
• hmotnostní spektrometrie MeSH
• interakce hostitele a patogenu genetika   MeSH
• karotenoidy biosyntéza   MeSH
• listy rostlin genetika   metabolismus   virologie   MeSH
• nemoci rostlin genetika   virologie   MeSH
• oxidace-redukce MeSH
• peroxid vodíku metabolismus   MeSH
• proteiny tepelného šoku klasifikace   genetika   metabolismus   MeSH
• Prunus avium virologie   MeSH
• regulace genové exprese u rostlin * MeSH
• rostlinné proteiny klasifikace   genetika   metabolismus   MeSH
• slivoň švestka virologie   MeSH
• tabák genetika   metabolismus   virologie   MeSH
• virus šarky švestky klasifikace   genetika   růst a vývoj   patogenita   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BRAFV600E mutations occur in ∼10% of colorectal cancer cases, are associated with poor survival, and have limited responses to BRAF/MEK inhibition with or without EGFR inhibition. There is an unmet need to understand the biology of poor prognostic BRAFMT colorectal cancer. We have used differential gene expression and pathway analyses of untreated stage II and stage III BRAFMT (discovery set: n = 31; validation set: n = 26) colorectal cancer, and an siRNA screen to characterize the biology underpinning the BRAFMT subgroup with poorest outcome. These analyses identified the unfolded protein response (UPR) as a novel and druggable pathway associated with the BRAFMT colorectal cancer subgroup with poorest outcome. We also found that oncogenic BRAF drives endoplasmic reticulum (ER) stress and UPR pathway activation through MEK/ERK. Furthermore, inhibition of GRP78, the master regulator of the UPR, using siRNA or small molecule inhibition, resulted in acute ER stress and apoptosis, in particular in BRAFMT colorectal cancer cells. In addition, dual targeting of protein degradation using combined Carfilzomib (proteasome inhibitor) and ACY-1215 (HDAC6-selective inhibitor) treatment resulted in marked accumulation of protein aggregates, acute ER stress, apoptosis, and therapeutic efficacy in BRAFMT in vitro and xenograft models. Mechanistically, we found that the apoptosis following combined Carfilzomib/ACY-1215 treatment is mediated through increased CHOP expression. Taken together, our findings indicate that oncogenic BRAF induces chronic ER stress and that inducers of acute ER stress could be a novel treatment strategy for poor prognostic BRAFMT colorectal cancer. Mol Cancer Ther; 17(6); 1280-90. ©2018 AACR.

• MeSH
• antitumorózní látky farmakologie   MeSH
• apoptóza účinky léků   genetika   MeSH
• biologické modely MeSH
• inhibitory proteinkinas farmakologie   MeSH
• kolorektální nádory farmakoterapie   genetika   metabolismus   mortalita   MeSH
• kyseliny hydroxamové farmakologie   MeSH
• lidé MeSH
• MAP kinasový signální systém MeSH
• mutace * MeSH
• nádorové biomarkery MeSH
• nádorové buněčné linie MeSH
• oligopeptidy farmakologie   MeSH
• prognóza MeSH
• proteiny tepelného šoku genetika   metabolismus   MeSH
• proteosyntéza MeSH
• protoonkogenní proteiny B-raf antagonisté a inhibitory   genetika   metabolismus   MeSH
• pyrimidiny farmakologie   MeSH
• signální dráha UPR účinky léků   MeSH
• signální transdukce účinky léků   MeSH
• stres endoplazmatického retikula účinky léků   genetika   MeSH
• transkripční faktor CHOP genetika   metabolismus   MeSH
• viabilita buněk účinky léků   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Lactococcus lactis is a food-grade lactic acid bacterium that is used in the dairy industry as a cell factory and as a host for recombinant protein expression. The nisin-controlled inducible expression (NICE) system is frequently applied in L. lactis; however new tools for its genetic modification are highly desirable. In this work NICE was adapted for dual protein expression. Plasmid pNZDual, that contains two nisin promoters and multiple cloning sites (MCSs), and pNZPolycist, that contains a single nisin promoter and two MCSs separated by the ribosome binding site, were constructed. Genes for the infrared fluorescent protein and for the human IgG-binding DARPin were cloned in all possible combinations to assess the protein yield. The dual promoter plasmid pNZDual enabled balanced expression of the two model proteins. It was exploited for the development of a single-plasmid inducible CRISPR-Cas9 system (pNZCRISPR) by using a nisin promoter, first to drive Cas9 expression and, secondly, to drive single guide RNA transcription. sgRNAs against htrA and ermR directed Cas9 against genomic or plasmid DNA and caused changes in bacterial growth and survival. Replacing Cas9 by dCas9 enabled CRISPR interference-mediated silencing of the upp gene. The present study introduces a new series of plasmids for advanced genetic modification of lactic acid bacterium L. lactis.

• MeSH
• antibakteriální látky farmakologie   MeSH
• CRISPR-Cas systémy MeSH
• editace genu metody   MeSH
• fermentace MeSH
• genetické inženýrství metody   MeSH
• genom bakteriální * MeSH
• guide RNA, Kinetoplastida genetika   metabolismus   MeSH
• imunoglobulin G genetika   metabolismus   MeSH
• klonování DNA MeSH
• Lactococcus lactis genetika   metabolismus   MeSH
• lidé MeSH
• methyltransferasy genetika   metabolismus   MeSH
• nisin farmakologie   MeSH
• plazmidy chemie   metabolismus   MeSH
• promotorové oblasti (genetika) účinky léků   MeSH
• proteiny tepelného šoku genetika   metabolismus   MeSH
• regulace genové exprese u bakterií * MeSH
• rekombinantní fúzní proteiny genetika   metabolismus   MeSH
• transgeny * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Hepatocellular carcinoma (HCC) has one of the worst prognoses amongst all malignancies. It commonly arises in patients with established liver disease and the diagnosis often occurs at an advanced stage. Genetic variations, such as single nucleotide polymorphisms (SNPs), may alter disease risk and thus may have use as predictive markers of disease outcome. The aims of this study were (i) to assess the association of two SNPs, rs430397 in GRP78 and rs738409 in PNPLA3 with the risk of developing HCC in a Sicilian association cohort and, (ii) to use a machine learning technique to establish a predictive combinatorial phenotypic model for HCC including rs430397 and rs738409 genotypes and clinical and laboratory attributes. The controls comprised of 304 healthy subjects while the cases comprised of 170 HCC patients the majority of whom had hepatitis C (HCV)-related cirrhosis. Significant associations were identified between the risk of developing HCC and both rs430397 (p=0.0095) and rs738409 (p=0.0063). The association between rs738409 and HCC was significantly stronger in the HCV positive cases. In the best prediction model, represented graphically by a decision tree with an acceptable misclassification rate of 17.0%, the A/A and G/A genotypes of the rs430397 variant were fixed and combined with the three rs738409 genotypes; the attributes were age, sex and alcohol. These results demonstrate significant associations between both rs430397 and rs738409 and HCC development in a Sicilian cohort. The combinatorial predictive model developed to include these genetic variants may, if validated in independent cohorts, allow for earlier diagnosis of HCC.

• MeSH
• alely MeSH
• frekvence genu MeSH
• genetická predispozice k nemoci genetika   MeSH
• genotyp MeSH
• hepatocelulární karcinom genetika   MeSH
• jednonukleotidový polymorfismus * MeSH
• kohortové studie MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipasa genetika   MeSH
• membránové proteiny genetika   MeSH
• nádory jater genetika   MeSH
• proteiny tepelného šoku genetika   MeSH
• rizikové faktory MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Sicilie MeSH

Planktothrix agardhii is one of the freshwater cyanobacteria that can produce the hepatotoxin microcystins (MC)-a real threat to human and animal health. Knowledge of the biological role of MC in producing organisms is highly desired to understand the driving force of MC production. Recently, emerging evidences have suggested that MC may have protective role in cells facing environmental stress. If this is true, one should expect differences in the cellular protective mechanisms between MC-containing and MC-deficient mutant strains. To test this hypothesis, it would be essential to investigate the consequences of the loss of MC in Planktothrix in the transcriptional responses of its heat shock proteins (Hsps) to abiotic stresses-an important component of cellular stress response. However, a crucial first step is prerequisite for the isolation of hsp genes here, as the genome of Planktothrix has not been fully published. Therefore, we have successfully isolated four hsp genes including clpC (hsp100), htpG (hsp90), groEL (hsp60), and groES (hsp10) from Planktothrix agardhii PCC 7805 using ramped annealing PCR (RAN-PCR) with consensus-degenerate hybrid oligonucleotide primers (CODEHOP) and annealing control primer (ACP) system. In addition, some putative regulatory sequences found in the upstream region of groESL operon of Planktothrix agardhii were also discussed.

• MeSH
• DNA bakterií chemie   genetika   MeSH
• fyziologický stres * MeSH
• klonování DNA * MeSH
• mikrocystiny metabolismus   MeSH
• molekulární sekvence - údaje MeSH
• proteiny tepelného šoku biosyntéza   genetika   MeSH
• sekvenční analýza DNA MeSH
• sinice genetika   metabolismus   fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The study describes the stress response in the central cotyledon zone of placental tissue and in maternal whole peripheral blood to pregnancy related complications including gestational hypertension (n = 31), preeclampsia w or w/o fetal growth restriction (n = 95), and fetal growth restriction (n = 39) using real-time RT-PCR and genes encoding Hsp27, Hsp60, Hsp70, Hsp90 and HspBP1 proteins. The placental tissue does not respond to pregnancy induced hypertension, fetal growth restriction and short-term severe preeclampsia that requires immediate termination of gestation. Upregulation of Hsp27, Hsp90 and HspBP1 appears just in case of long-term deteriorated conditions (usually in mild preeclampsia, that enable further continuation of gestation, when properly treated). On the other hand, maternal circulation is able to reflect both maternal and fetal pathologic conditions. While pregnancy related complications always induce upregulation of Hsp70 and downregulation of Hsp90 in maternal whole peripheral blood, the increase of Hsp60 mRNA levels occurs entirely in patients with preeclampsia and/or fetal growth restriction. Hsp60, Hsp70 and Hsp90 are dysregulated in maternal circulation irrespective of the severity of the disease (in both mild and severe preeclampsia) and the requirements for the delivery (before and after 34th week of gestation). Nevertheless, the highest Hsp60 mRNA levels may be observed in pregnancies with signs of the centralization of the fetal circulation associated with fetal hypoxia.

• MeSH
• dospělí MeSH
• fyziologický stres genetika   MeSH
• komplikace těhotenství diagnóza   genetika   metabolismus   patofyziologie   MeSH
• krevní tlak MeSH
• lidé MeSH
• messenger RNA * MeSH
• placenta metabolismus   MeSH
• preeklampsie diagnóza   genetika   metabolismus   patofyziologie   MeSH
• proteiny tepelného šoku genetika   MeSH
• regulace genové exprese MeSH
• retrospektivní studie MeSH
• růstová retardace plodu diagnóza   genetika   metabolismus   MeSH
• stupeň závažnosti nemoci MeSH
• těhotenství MeSH
• ultrasonografie dopplerovská barevná MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Insect adipokinetic hormones (AKHs) are pleiotropic hormones known to play a protective role in response to oxidative stress (OS). However, the precise signaling pathways are unclear. We present evidence that AKH may primarily employ the Forkhead box class O transcription factor (FoxO) to exert this effect. The impact of knocking down AKH synthesis or its over-expression in its response to OS was studied in Drosophila melanogaster. AKH knockdown (AKH-RNAi) as well as AKH overexpression (AKH-oex) was achieved using the Gal-4/UAS system and controls were w(1118) (+/+), AKH-Gal4/+, UAS-AKH/+ and UAS-AKH-RNAi/+. Exposure to 80 μM hydrogen peroxide (HP) revealed that AKH-RNAi flies showed significantly higher mortality than AKH-oex or the respective control lines. This susceptibility was evidenced by significantly enhanced levels of protein carbonyls - a biomarker of OS, in AKH-RNAi flies compared to controls and AKH-oex flies. Interestingly, AKH-oex flies had the least amount of protein carbonyls. AKH-RNAi flies had significantly less dFoxO transcript and translated protein compared to control and AKH-oex flies in un-challenged condition as well as when challenged with HP. Sestrin - a major antioxidant defense protein and one of the targets of dFoxO - was also significantly down-regulated (both at mRNA and protein level) in AKH-RNAi flies (both unchallenged and challenged with HP) compared to control flies and flies with over-expressed AKH. These findings imply that dFoxO may act downstream of AKH as a transcription factor to mediate response to OS in D. melanogaster.

• MeSH
• biologické markery metabolismus   MeSH
• Drosophila melanogaster genetika   fyziologie   MeSH
• forkhead transkripční faktory genetika   metabolismus   MeSH
• geneticky modifikovaná zvířata MeSH
• hmyzí hormony antagonisté a inhibitory   genetika   metabolismus   MeSH
• karbonylace proteinů účinky léků   MeSH
• křížení genetické MeSH
• kyselina pyrrolidonkarboxylová analogy a deriváty   antagonisté a inhibitory   metabolismus   MeSH
• léková rezistence MeSH
• MAP kinasový signální systém účinky léků   MeSH
• messenger RNA metabolismus   MeSH
• oligopeptidy antagonisté a inhibitory   genetika   metabolismus   MeSH
• oxidační stres * MeSH
• oxidancia toxicita   MeSH
• peroxid vodíku toxicita   MeSH
• proteiny Drosophily antagonisté a inhibitory   genetika   metabolismus   MeSH
• proteiny tepelného šoku genetika   metabolismus   MeSH
• regulace genové exprese * účinky léků   MeSH
• rekombinantní proteiny chemie   metabolismus   MeSH
• RNA interference MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH