Fibromyxoid aSoft Tissue Tumor With PLAG1 Fusion-The First Case in an Adult Patient.

With the expanding possibilities of human genome research in recent years, the number of cases of soft tissue tumors that we are able to classify into the correct subgroups and to reveal their molecular profile is increasing. Among such tumors, we can also consider neoplasms that have a specific fusion of genes, in our case namely the pleomorphic adenoma gene 1 (PLAG1) and its partner. PLAG1 gene fusions were previously associated mainly with salivary gland pleomorphic adenomas, lipoblastomas, myoepithelial tumors, uterine epitheloid, myxoid leiomyosarcomas, and, recently, with PLAG1-rearranged fibromyxoid soft tissue tumors.

Qualitative and quantitative analysis of lipid droplets in mature 3T3-L1 adipocytes using oil red O.

By differentiating into mature adipocytes, 3T3-L1 cells can be utilized as a model cell line to investigate (pre)adipocyte function in vitro. Here, we present a protocol for combining qualitative and quantitative analysis of lipid droplets in mature 3T3-L1 adipocytes using oil red O. We describe steps to differentiate 3T3-L1 preadipocytes to adipocytes and give detailed procedures to determine total lipid amount as well as lipid droplet size and number using microscopic devices and an ImageJ macro.

Dysfunction of the adhesion G protein-coupled receptor latrophilin 1 (ADGRL1/LPHN1) increases the risk of obesity.

Obesity is one of the diseases with severe health consequences and rapidly increasing worldwide prevalence. Understanding the complex network of food intake and energy balance regulation is an essential prerequisite for pharmacological intervention with obesity. G protein-coupled receptors (GPCRs) are among the main modulators of metabolism and energy balance.

Novel and unusual fusion partners in aneurysmal bone cyst and their role in pathogenesis and histopathological evaluation of this disease.

Aims: The purpose of this study is to report novel and unusual fusion partners in aneurysmal bone cysts (ABCs). These findings may be useful in routine diagnostics as well as in studying the biology of -related disorders.

Methods: A cohort of seven patients diagnosed with ABC examined between 2014 and 2023 at Motol University Hospital in Prague was included into this retrospective non-randomised study.

Clinical and molecular study of radiation-induced gliomas.

In this study, we provide a comprehensive clinical and molecular biological characterization of radiation-induced gliomas (RIG), including a risk assessment for developing gliomas. A cohort of 12 patients who developed RIG 9.5 years (3-31 years) after previous cranial radiotherapy for brain tumors or T-cell acute lymphoblastic leukemia was established.

Mazabraud's syndrome: A case report supported by molecular studies and review of the literature.

Mazabraud's syndrome represents rare benign disorder characterized by simultaneous occurrence of fibrous dysplasia of bone and intramuscular myxomas within surrounding soft tissue. Mutations of gene were proven to be causative for this condition. Here, we present a case report of a patient with unusual manifestation of this disease, who developed a pathological fracture of the femur in the setting of monostotic fibrous dysplasia.

A rational approach to the CNS tumors diagnostics.

The new WHO classification of CNS tumors is largely based on molecular diagnostic. Without molecular methods some entities can no longer be diagnosed. We are trying to show a rational approach to the CNS tumors diagnostics, which is based on conventional molecular methods such as RT-PCR, Sanger sequencing, MLPA, extended by the next generation sequencing (NGS) and methylation SNP array.

High-throughput workflow for identification of phosphorylated peptides by LC-MALDI-TOF/TOF-MS coupled to in situ enrichment on MALDI plates functionalized by ion landing.

We report an MS-based workflow for identification of phosphorylated peptides from trypsinized protein mixtures and cell lysates that is suitable for high-throughput sample analysis. The workflow is based on an in situ enrichment on matrix-assisted laser desorption/ionization (MALDI) plates that were functionalized by TiO2 using automated ion landing apparatus that can operate unsupervised. The MALDI plate can be functionalized by TiO2 into any array of predefined geometry (here, 96 positions for samples and 24 for mass calibration standards) made compatible with a standard MALDI spotter and coupled with high-performance liquid chromatography.

Lateral resolution in NALDI MSI: back to the future.

Nanostructure-assisted laser desorption/ionization (NALDI) has been recognized as a powerful matrix-free mass spectrometry tool ideal for imaging of small molecules. In this report, the NALDI approach was compared with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry in terms of sensitivity, reproducibility, and lateral resolution, which can be achieved in mass spectrometry imaging (MSI) experiments using a Nd:YAG laser. Scanning electron microscopy was used for surface topology analysis and evaluation of a putative surface-enhanced sensitivity effect, which was observed upon reduction of the laser focus.

In-situ enrichment of phosphopeptides on MALDI plates modified by ambient ion landing.

We report substantial in-situ enrichment of phosphopeptides in peptide mixtures using titanium and zirconium dioxide-coated matrix assisted laser desorption-ionization (MALDI) plates prepared by recently reported ambient ion landing deposition technique. The technique was able to modify four common materials currently used for MALDI targets (stainless steel, aluminum, indium-tin oxide glass and polymeric anchor chip). The structure of the deposited dioxide was investigated by electron microscopy, and different surfaces were compared and discussed in this study.

Time-dependent oxidation during nano-assisted laser desorption ionization mass spectrometry: a useful tool for structure determination or a source of possible confusion?

This work reports on a new and extremely simple approach for determination of a double bond position by a laser desorption ionization mass spectrometry. It is solely based on the catalytic properties of nanostructured surfaces used in nanoassisted laser desorption ionization experiments. These surfaces can induce oxidation of analytes, which results in a mass shift that can be detected by mass spectrometry.

In vivo behaviour of low-temperature calcium-deficient hydroxyapatite: comparison with deproteinised bovine bone.

This study aims to evaluate in detail the biological osteoconductive properties of the low-temperature synthetic porous calcium-deficient hydroxyapatite and to compare it with the biological apatite. Bone reactions to granules of similar sizes of the low-temperature hydroxyapatite and commercially available non-sintered deproteinized bovine bone were compared. Two different temperatures were used to fabricate two batches of newly developed porous hydroxyapatite with different carbonate groups content and specific surface area.

Occurrence of lipids in the liver of the hypertriglyceridemic rats.

Aims: The purpose of this study was to demonstrate the accumulation and distribution of lipids in the liver of the adult Prague hereditary hypertriglyceridemic (HHTg) rats. They reveal an increased expression of 11beta-hydroxysteroid dehydrogenase 1 (11HSD1), which locally increases concentration of corticosterone in the liver. We studied the effect of the 11HSD1 inhibition on the lipid content.

Cell viability and protein turnover in nongrowing Bacillus megaterium at sporulation suppressing temperature.

In Bacillus megaterium, a temperature that suppresses sporulation (43 degrees C) only slightly exceeds both the optimum growth temperature and the temperature still permitting sporulation (40-41 degrees C). Here we show that, when cells grown at 35 degrees C and transferred to a sporulation medium, were subjected to shifts between 35 degrees C and the sporulation suppressing temperature (SST, 43 degrees C), their development and proteolytic activities were deeply affected. During the reversible sporulation phase that took place at 35 degrees C for 2-3 h (T2-T3), the cells developed forespores and their protein turnover was characterized by degradation of short-lived proteins and proteins made accessible to the proteolytic attack because of starvation.

In vivo and In vitro function of the intracellular proteolytic apparatus in nongrowing bacillus megaterium under heat stress.

In Bacillus megaterium sporulating at 35 degreesC, up to 90% of 10-min pulse-labeled proteins were degraded. Degradation proceeded in two waves. Short-lived proteins, i.

Asporogenic Bacillus megaterium mutant 27-36 degrades intrinsically short-lived proteins but fails to convert most of other proteins to a short-lived fraction.

Asporogenic mutant blocked in the 0-II sporulation stage degraded pulse-labelled proteins in the sporulation medium at the same rate as the parental strain for the first two hours. The degraded fraction was mostly composed of intrinsically short-lived proteins which were degraded even after enriching the medium with amino acids and growth resumption. Proteins accessible to degradation because of nutritional shift down formed a lesser proportion of this fraction.

Determination of platinum in plants by emission spectrometry after preconcentration on modified silicagel.

Silicagel Separon SGX C18 (particle size 7 microm) was suitable for the preconcentration of 2-20 microg of Pt from 0.1M hydrochloric acid in the presence of cationic surfactants especially dimethyllaurylbenzylammonium bromide, with subsequent elution with 96% ethanol. The recovery was 86-110% for 2 microg of Pt.

External factors involved in the regulation of synthesis of an extracellular proteinase in Bacillus megaterium: effect of temperature.

We studied the effect of temperature on the production of an extracellular neutral metalloproteinase of Bacillus megaterium in a laboratory fermentor under constant aeration and pH. The optimal temperature for growth (35-38° C) was higher than that for the synthesis of proteinase during exponential growth (below 31° C). The critical biomass concentration at which the exponential growth terminated decreased with increase in cultivation temperature.

Effect of actinomycin D on viability, sporulation and nucleotide pool of Bacillus megaterium.

A transient 7-fold rise of ppGpp concentration, 2-3-fold increase of pppGpp concentration and 50% drop of the concentration of GTP in Bacillus megaterium cells immediately after their transfer to the sporulation medium were observed. Actinomycin D, in concentrations inhibiting RNA synthesis by 95%, blocked the rise of the (p)ppGpp pool and caused an instant several-fold increase of the GTP level. When the cells were exposed to actinomycin D in the sporulation medium for a 1-h period (time 0-1 h, 1-2 h or 2.

Regulation of extracellular proteins and alpha-amylase secretion by temperature in Bacillus subtilis.

alpha-Amylase was found to be the main protein secreted by Bacillus subtilis, corresponding to 90, 87 and 60% of total extracellular proteins at 30, 40 and 45 degrees C, respectively. A change in temperature can affect the pattern of proteins secreted as detected by gel electrophoresis. 14C-Leucine incorporation into extracellular proteins and their proportion at the end of the growth phase was higher at 30 degrees C than that at 40 or 45 degrees C.

Effect of temperature on alpha-amylase formation and DNA replication in Bacillus subtilis.

The effect of temperature on extracellular alpha-amylase synthesis and chromosomal and plasmid DNA replication in Bacillus subtilis A18 carrying plasmid pMI10 was studied. The specific growth rate mu increased with elevated temperature up to 42.5 degrees C, while the activities of alpha-amylase per population dry mass decreased.

Plasmid pIMI38--the pBR322 derivative with increased stability in E. coli cells.

Plasmid pIM138 which had been characterized by the higher resistance of its DNA replication to the action of clorobiocin in comparison with the progenitor plasmid, was tested for its stability in host cells in the absence of the antibiotic. Growing without selective pressure, pIM138 was better maintained in cells than pBR322. The stability in the presence and in the absence of clorobiocin can be unanimously assigned to the plasmid itself, but some influence of host cells cannot be excluded.

Turnover of abnormal proteins in Bacillus megaterium and Saccharomyces cerevisiae: differences between in vivo and in vitro degradation.

Degradation of abnormal proteins in Bacillus megaterium and Saccharomyces cerevisiae in vivo was compared with that in cell-free extracts. Protein degradation in vivo, when the cells were labelled with 14C-leucine during growth in the presence of ethionine, was affected by the concentration of the analogue used. Proteins synthesized in the presence of 0.

[Succinate dehydrogenase activity in the mesonephros in the process of development of chick embryos].

The activity of succinate dehydrogenase in the mesonephros tissue of chick embryos is rather high on the 6th day of embryonal development and is not subjected to significant changes up to the 17th day. The absence of wide variations in the activity of this enzyme during the period of degeneration of the mesonephros may be due to a parallel decrease in the organ's mass and in the number of its mitochondria.