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This is a case report of B-cell non-Hodgkin lymphoma coexisting with parathyroid carcinoma as an unexpected cause of hypercalcemia. Hypercalcemia is a common accompanying paraneoplastic phenomenon. Primary hyperparathyroidism less frequently leads to increased levels of plasma calcium in malignant diseases. Simultaneous or double malignancies, represented by parathyroid carcinoma in this case, are extremely rare. Parathyroid carcinoma may be suspected based on severe clinical signs, significant laboratory findings, and the tumor size.

This article analyses the rise of a new social formation, the project class which represents a recent complex societal and political change. It is argued that project proliferation along with reforms of the administrative structures, changes in the nature of developmental policies and the increasing importance of cultural and cognitive elements of territorial development are the driving forces behind the emergence of a new class. This article analyses the mediatory position of the new social class in the redistribution of public and particularly private development funds, and in the transfer of materials, ideas, knowledge and power. Special attention is given to the intellectual capital that provides legitimacy for the project class and to the flexible social and economic positioning by which the members of the new class create, use and consume intellectual and material products of the information society in order to be competitive on the markets of the project-based economy. The article apprehends the relationship between the emergence of the project class and the transformation of the local power structure. Finally, it demonstrates the challenges related to new forms of knowledge use associated with the emergence of the new social class.

Thick Film Technology (TFT) offers a new platform for analytical procedures in Electrochemistry. The most routine technology is screen printing. However, it can introduce new procedures connected with miniaturisation or combination of microfluidic and electrodes. TFT use in electrochemistry is discussed. Examples of different sensors are demonstrated. Details are referred to in the original literature. Advanced applications combining TFT with other technologies are demonstrated (capillary electrophoresis on a chip and a sensor with integrated heating and thermometer). Future of TFT in electrochemistry is discussed, also the drawbacks, connection to production and commercial application are noticed.

ABSTRACTListeriosis is a serious foodborne infection that disproportionately affects elderly adults, pregnant women, newborns, and immunocompromised individuals. Diagnosis is made by culturing Listeria monocytogenesfrom sterile body fluids or from products of conception. This report describes the investigations of two listeriosis pseudo-outbreaks caused by contaminated laboratory media made from sheep blood.

The interactions of pepsin with immobilized trivalent metal ions and the participation of the enzyme phosphate group in this process were investigated using high performance immobilized metal affinity chromatography. Two different sorbents were used: the newly prepared one, consisting of Ga3+ chelate of (6‐amino‐1‐hydroxyhexane‐1,1‐diyl) bis(phosphonic acid) covalently bound to a methacrylate support (BP‐Ga3+), and the commercial one, containing immobilized Fe3+ ions (POROS MC20‐Fe3+). The comparison of the behavior of porcine pepsin A and its partially dephosphorylated form on both sorbents showed that both forms of pepsin were adsorbed under the same conditions. To eliminate the participation of free carboxyl groups in pepsin adsorption, both enzyme forms were modified by amidation or esterification. Native enzyme and its partially dephosphorylated form both with modified carboxyl groups differed in their interaction with immobilized Ga3+ and Fe3+. Phosphorylated pepsin molecules with esterified carboxyl groups were adsorbed on both sorbents while nonphosphorylated ones with esterified carboxyl groups were not adsorbed.

ABSTRACTA three-step method for the purification of Enterocytozoon bieneusispores from stool specimens was developed. The primary process of purification of the spores from bacterial contaminants involved Percoll gradient centrifugation followed by additional separation using cesium chloride density gradient centrifugation. The cesium chloride-isolated spores were further purified using a flow cytometer with cell sorting capabilities. Sorting was performed without the use of antibodies, fluorochromes, or dyes, leaving the sorted spores in their native state, which appears to be less destructive for spores. When quantified by flow cytometry using tubes with known numbers of highly fluorescent polystyrene beads, the sorted material showed a slight decrease in light scatter characteristics compared with the slightly larger Encephalitozoonspecies spores. Although the overall recovery of the E. bieneusispores was low, calcofluor and Gram chromotrope staining, indirect immunofluorescence assay, and transmission electron microscopy revealed that the sorted material was highly purified and contained large numbers of E. bieneusispores and relatively few bacteria and other debris. The sorted material appeared to be sufficiently pure and could be used for in vitro culture and for the development of a variety of diagnostic reagents as well as in studying the genome of E. bieneusiand host-parasite interactions.

Nonsteroidal antiandrogens (AA) cannot be topically used for androgenetic alopecia (AGA) because of systemic resorption. A new class of androgen receptor (AR) suppressors designed for safe topical treatment of AGA was synthesized from (3-amino-2-hydroxy-2-methyl-N-(4-nitro-3-trifluoromethyl)phenyl) propanamide (BP-34), to contain perfluoroalkyl moieties. The trifluoromethyl derivative (fluridil) at 10 μM decreased expression of the AR in LNCaP human cells by 95%, its serum half-life was 6 h; it decomposes hydrolytically to BP-34 and trifluoroacetic acid. Acute intraperitoneal maximum tolerated dose (MTD) of fluridil in mice is 270–300 mg/kg/d and the subacute MTD is 450 mg/kg/d. The oral LD50 in mice was 2,872 mg/kg in males, 2,232 mg/kg in females, and >2,500 mg/kg in rats. Fluridil solution in isopropanol was not cutaneously absorbed in rabbits, did not sensitize or show any phototoxic or photoallergic effects on guinea pig skin, and demonstrated no skin irritation potential in rabbits and humans. Fluridil solid induced only slight and reversible eye irritancy in rabbits and displayed no cytotoxicity to rabbit corneal fibroblasts in vitro. Fluridil demonstrated no significant mutagenicity potential by Ames method. In a double-blind study, 43 males with AGA, Norwood grade II to Va, used topical 2% fluridil in isopropanol or the vehicle daily for 12 months. Anagens (growing hairs) increased in the fluridil group from 76% to 89%. All hematological and biochemistry values remained within normal range, including testosterone, which varied but seasonally. No fluridil or its decomposition product (BP-34) was detected in serum. No adverse side effects were reported. Drug Dev. Res. 59:292–306, 2003. © 2003 Wiley-Liss, Inc.

Transformation of polychlorinated biphenyls was studied using different strains of in vitro cultured cells of horseradish (Armoracia rusticana L.). Time and concentration dependence of this process and production of intracellular and extracellular peroxidases were measured with differentiated shooty teratoma culture K54. The yield of PCB transformation and the viability of the cells were highly dependent on PCB concentration. 100 ppm PCB totally inhibited growth of the cells, and reduced their metabolism of xenobiotics. Experiments with a peroxidase (POX) inhibitor, propylgallate, and a cytochrome P450 inhibitor, aminobenztriazole, indicated the involvement of both enzymatic systems in PCB metabolism.

ABSTRACTInfection with a replication-competent bovine leukemia virus structural gene vector (BLV SGV) is an innovative vaccination approach to prevent disease by complex retroviruses. Previously we developed BLV SGV that constitutively expresses BLV gag, pol, and envand related cis-acting sequences but lacks tax, rex, RIII, andGIVand most of the BLV long terminal repeat sequences, including the cis-acting Tax and Rex response elements. The novel SGV virus is replication competent and replicates a selectable vector to a titer similar to that of the parental BLV in cell culture. The overall goal of this study was to test the hypothesis that infection with BLV SGV is nonpathogenic in rabbits. BLV infection of rabbits by inoculation of cell-free BLV or cell-associated BLV typically causes an immunodeficiency-like syndrome and death by 1 year postinfection. We sought to evaluate whether in vivo transfection of BLV provirus recapitulates pathogenic BLV infection and to compare BLV and BLV SGV with respect to infection, immunogenicity, and clinical outcome. Three groups of rabbits were subjected to in vivo transfection with BLV, BLV SGV, or negative control DNA. The results of our 20-month study indicate that in vivo transfection of rabbits with BLV recapitulates the fatal BLV infection produced by cell-free or cell-associated BLV. The BLV-infected rabbits exhibited sudden onset of clinical decline and immunodeficiency-like symptoms that culminated in death. BLV and BLV SGV infected peripheral blood mononuclear cells and induced similar levels of seroconversion to BLV structural proteins. However, BLV SGV exhibited a reduced proviral load and did not trigger the immunodeficiency-like syndrome. These results are consistent with the hypothesis that BLV SGV is infectious and immunogenic and lacks BLV pathogenicity in rabbits, and they support the use of this modified proviral vector delivery system for vaccines against complex retroviruses like BLV.

We studied the effect of temperature on the production of an extracellular neutral metalloproteinase of Bacillus megateriumin 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. The specific rate of proteinase synthesis decreased when the critical biomass concentration was achieved. The observed decrease in proteinase synthesis was related to the cultivation temperature. The temperature also influenced the level of mRNA coding for proteinase. We formulated a mathematical model of cultivation describing the dependence of growth and proteinase synthesis on dissolved oxygen and temperature. The parameters of the model were identified for temperature intervals from 21 to 41° C using a computer. The optimum temperature for the enzyme production was 21° C. The productivity (enzyme activity/time) was maximal at 24–28° C. When optimizing the temperature profile of cultivation, we designed a suboptimal solution represented by a linear temperature profile. We have found that under conditions of continuous decrease in temperature, the maximal production of the proteinase was achieved at a broad range of temperature (26–34° C) when the rate of temperature decrease was 0.2–0.8° C/h. The initial optimal temperature for the enzyme productivity was in the range of 32–34° C. The optimum temperature decrease was 0.8° C/h.

Genetically simplified derivatives of complex retroviruses that replicate in animal models are useful tools to study the role of the complex regulatory genes in virus infection and pathogenesis and were proposed as a novel approach toward the development of vaccines against complex retroviruses. Previously we developed genetically simple derivatives of bovine leukemia virus (BLV) that can replicate in tissue culture independently of the BLV regulatory proteins, Tax and Rex, and the RIII and GIV open reading frames (K. Boris-Lawrie and H. M. Temin, J. Virol. 69:1920-1924, 1995). These derivatives are encoded on novel, hybrid retrovirus genomes that contain transcriptional control sequences of a simple retrovirus and gag-pol or env genes of the complex BLV. The first-generation simple BLV derivatives replicate as complementary viruses (coviruses) by using separate gag-pol or env genomes, and therefore virus spread is limited to cells that are infected with both covirus genomes. Here we describe a second-generation simple BLV derivative that is encoded on a single hybrid genome. We show the virus to be replication competent by successive passage on D17 target cells and by analysis of viral RNA and proteins in the infected cells. Furthermore, we evaluate the immunogenicity and infectivity of the simple BLV derivatives in a BLV animal model. Small groups of rats were injected either with virus-producing cells or with proviral DNA. Western immunoblot analysis revealed that antibodies against the major viral antigenic determinants are induced in response to either method of introduction and that seroconversion is sustained in most of the rats for at least 6 months (the duration of the study). The magnitudes of the antiviral responses were similar in rats infected with the first-generation simple BLV coviruses, the second-generation replication-competent derivative, or wild-type BLV. Wild-type BLV typically infects peripheral blood mononuclear cells (PBMC), and the simple BLV derivatives were also found to infect PBMC as demonstrated by PCR amplification of proviral sequences and reverse transcriptase PCR amplification of viral RNA in treated rats. These results establish that simple BLV derivatives lacking tax and rex are infectious and immunogenic in rats. These viruses will be useful tools in comparative studies with BLV to evaluate the role of tax and rex in maintenance of virus load and in disease outcome.

The paper summarizes 208 heat flow values that have been collected in former Czechoslovakia (152 of them in Czech Republic, and 56 in Slovakia). The mean heat flow and its standard deviation for the whole region is 71.0 ± 22.8 mW·m2. To interpret the regional geothermal activity, the results of similar data from the adjacent part of Central Europe were incorporated into the general picture. The heat flow map was correlated with the crustal thickness pattern, with the radiogenic heat production and the characteristic crustal temperature-depth profiles were constructed. Geothermal data were used to assess the mantle (Moho) heat flow and the thickness of the thermal lithosphere. Special attention was paid to the possible transient component of the heat flow due to the Tertiary-Quaternary volcanism in the northern part of the Bohemian Massif.

Thyroid abscess is a rare condition with a dramatic clinical course. Most commonly, thyroid abscess develops in a multinodular goiter. Rarely, the infection is iatrogenic. This report describes an abscess developing after fine-needle aspiration biopsy. The infectious agent was Escherichia coli, not a typical skin pathogen. The patient's condition required intubation and temporary tracheostomy. The case underlines the necessity of adhering to strict aseptic technique even in minimally invasive procedures.

Heat and thermo-mechanical treatments with various processing parameters were applied to middle carbon low alloyed 42SiMn steel. The aim of the treatment was to obtain multiphase microstructure typical for TRIP (Transformation induced plasticity) steel and to achieve the best combination of ultimate tensile strength and ductility. TRIP steels typically possess about 5-15% of metastable retained austenite, which can transform to martensite during plastic deformation. The gradual phase transformation during loading postpones the onset of necking, thus increasing ultimate tensile strength and ductility at the same time. Manganese and silicon, used as the main alloying elements of the experimental steel, are employed to increase austenite stability and to hinder cementite precipitation during the treatment. All proposed methods of heat and thermo-mechanical treatment contain bainitic hold at 400 degC or 425 degC. The final microstructures were very complex, consisting of bainite, ferrite, very small areas of extremely fine perlite lamellas, about 10% of retained austenite and M-A constituent (austenitic islands partially transformed to martensite). Even though pearlite and martensite are undesirable microstructure in TRIP steel, the tensile strength ranged from 850 to 1065 MPa and ductility A5mm from 26 to 47 %.

The paper reports on the behaviour of low alloyed TRIP (transformation induced plasticity) steel with Niobium during tensile test. The structures were analysed using in-situ tensile testing coupled with electron backscattering diffraction (EBSD) analysis carried out in scanning electron microscope (SEM). Steel specimens were of same chemical composition; however three different annealing temperatures, 800 degC, 850 degC and 950 degC, were applied to the material during the heat treatment. The treatment consisted of annealing for 20 minutes in the furnace; cooling in salt bath after the heating and holding at 425 degC for 20 minutes for all the samples. Untreated bar was used as reference material. Flat samples for deformation stage were cut out of the heat-treated bars. In situ documentation of microstructure and crystallography development were carried out during the deformation experiments. High deformation lead to significant degradation of EBSD signal.

The contemporary development of technological processes for the production of modern multiphase steels can be characterized by the need for precise control of their technological parameters. The design of modern technological processes that allow sophisticated microstructures to be obtained usually cannot be carried out on real production equipment for technical as well as economical reasons. Therefore, new processes and test devices are continuously being developed to make it possible to simulate and model thermomechanical treatments on small specimens with precise control and monitoring of process parameters. A simulator for experimental modelling of thermomechanical processes has been developed at the University of West Bohemia. In this paper, to demonstrate the feasibility of simulating thermomechanical treatments with this setup on a lab scale, we discuss the thermomechanical treatment of TRIP steels without isothermal holding - a processing route that is difficult to handle and thus poses several technological as well as economic problems. The realistic processing of wire rolling with different cooling strategies is tested on the TRIP CMnSiNb steel. Our results show that the processing route without isothermal holding allows to obtain multiphase microstructures with a tensile strength of up to 835 MPa and a ductility A5mm = 25%.

No relevant conflicts of interest to declare.

Constantinescu: Shire: Consultancy, Speakers Bureau; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Personal Genetics: Membership on an entity's Board of Directors or advisory committees; Amgen: Speakers Bureau.