SERS of cylindrospermopsin cyanotoxin: Prospects for quantitative analysis in solution and in fish tissue.

Cylindrospermopsin (CYN), a cyanotoxin occurring in environmental waters as a cyanobacteria metabolite, has recently raised increased interest both in the scientific community and the environmental, food control and health care bodies due to the incidence of poisoning reports and the lack of prompt, effective detection and monitoring techniques. Here we report comprehensive Raman and SERS spectroscopy data on CYN cyanotoxin and provide a detailed characterization of the vibrational Raman signal based on DFT calculation as well as the adsorption properties with respect to the silver nanoparticles surface. Quantitative SERS analysis was achieved for concentrations range from 0.218 nM to 2.18 µM in aqueous solution. We further investigated the SERS discrimination of artificially intoxicated fish tissue from normal one, using linear discriminant analysis. Significant changes in SERS signal of toxic tissue compared to normal one allowed clear and fast differentiation of toxic tissue with 100% specificity/sensitivity. The cross-validation procedure provided 100% clear separation based on the SERS data. The results open reliable perspectives for SERS monitoring the environmental water bodies.

212Bi-212Po alpha source for the calibration and functional testing of silicon detectors: Preparation and characterisation.

Silicon detectors are widely used in space radiation dosimeter systems for measuring energetic charged particles. Calibration of such systems is usually performed with protons and heavier ions in high energy particle accelerators. For preliminary energy calibration and functional testing of silicon detectors, at any time during the development, an equipment producing a thin 212Bi-212Po alpha particle emitting source was designed and constructed. Our aim was to develop an alpha source with negligible self-shielding and short life-time in order to prevent the long-term contamination of the detectors with alpha particle emitting nuclides. In the present paper, a description of the method chosen, and the equipment developed are given. Estimates of the activity of the source produced was obtained from measurements with the RADTEL space radiation telescope under development in the Centre for Energy Research, Hungarian Academy of Sciences (MTA EK). It was also used to verify that the alpha particle emitting source is suitable for the preliminary calibration and functional testing of silicon detector systems.

The Epithelial-Mesenchymal Transition Pathway in Two Cases with Gastric Metastasis Originating from Breast Carcinoma, One with a Metachronous Primary Gastric Cancer.

BACKGROUND: Metastases to the stomach are extremely rare and the metastatic pathway is not well understood. OBJECTIVE: To present two unusual gastric metastases and a review of the literature regarding the pathway of Epithelial Mesenchymal Transition (EMT) in the metastatic cells. METHOD: The clinicopathological aspects of the two cases were presented in the light of the most recent patents. Data about patents were obtained from the online databases PubMed, World Intellectual Property Organization (WIPO) and Google patents. RESULTS: In the first case, in a 73-year-old female, total gastrectomy was performed for a Gastric Cancer (GC) that was proved to be, based on the immunohistochemical features (positivity for mammaglobin and estrogen receptor and negativity for E-cadherin, ß-catenin, CD44 and maspin), a metastasis from an invasive lobular carcinoma of the breast, that was later confirmed. In the second case, a 67-year-old female with invasive ductal carcinoma of the breast, which benefited from chemotherapy and mastectomy, presented a metachronous gastric adenocarcinoma with collision-type metastatic breast ductal carcinoma. The aggressiveness of the GC cells was induced through the E-cadherin/maspin pathway, while the CD44-related stem-like properties of the tumor cells induced the aggressiveness of ductal carcinoma. CONCLUSION: In females with breast cancer, a possible metastasis in the stomach should be taken into account. Maspin and VSIG1 are not involved in breast cancer histogenesis. The Wnt/ß-catenin signaling is not involved in the lobular carcinoma progression. The CD44/HER2 positivity in ductal carcinoma cells might indicate high risk of distant metastasis and low response to chemotherapy.

CALIBRATION OF A WHOLE BODY COUNTER FOR 241Am WITH THE LLNL CHEST PHANTOM.

An occupational incorporation event occurred at the Radioactive Waste Treatment and Disposal Facility in December 2013 at Püspökszilágy, Hungary. Internal contamination due to (241)Am was discovered by a regular routine whole body counting measurement at the Centre for Energy Research, Hungarian Academy of Sciences. After that, a whole body counter was calibrated for an organ counting geometry. For preliminary calibration, a home-made MIX-D chest phantom was applied simulating uniform lung activity distribution by (241)Am point sources located in different positions within the lung volume of the phantom. In order to carry out a more precise calibration, a Lawrence Livermore National Laboratory (LLNL) chest phantom was provided by the International Atomic Energy Agency. For counting efficiency over the lungs, values of 0.46±0.19 and 0.55±0.07 cps kBq(-1) were obtained for the MIX-D and the LLNL phantom, respectively; thus, the results are in good agreement.

Dose measurements in space by the Hungarian Pille TLD system.

Exposure of crew, equipment, and experiments to the ambient space radiation environment in low Earth orbit poses one of the most significant problems to long-term space habitation. Accurate dose measurement has become increasingly important during the assembly (extravehicular activity (EVA)) and operation of space stations such as on Space Station Mir. Passive integrating detector systems such as thermoluminescent dosemeters (TLDs) are commonly used for dosimetry mapping and personal dosimetry on space vehicles. The well-known advantages of passive detector systems are their independence of power supply, small dimensions, high sensitivity, good stability, wide measuring range, resistance to environmental effects, and relatively low cost. Nevertheless, they have the general disadvantage that for evaluation purposes they need a laboratory or large--in mass and power consumption--terrestrial equipment, and consequently they cannot provide time-resolved dose data during long-term space flights. KFKI Atomic Energy Research Institute (KFKI AEKI) has developed and manufactured a series of thermoluminescent dosemeter systems for measuring cosmic radiation doses in the 10 microGy to 10 Gy range, consisting of a set of bulb dosemeters and a compact, self-contained, TLD reader suitable for on-board evaluation of the dosemeters. By means of such a system, highly accurate measurements were carried out on board the Salyut-6, -7 and Mir Space Stations as well as on the Space Shuttle. A detailed description of the system is given and the comprehensive results of these measurements are summarised.

In vitro cell irradiation systems based on 210Po alpha source: construction and characterisation.

One way of studying the risk to human health of low-level radiation exposure is to make biological experiments on living cell cultures. Two 210Po alpha-particle emitting devices, with 0.5 and 100 MBq activity, were designed and constructed to perform such experiments irradiating monolayers of cells. Estimates of dose rate at the cell surface were obtained from measurements by a PIPS alpha-particle spectrometer and from calculations by the SRIM 2000, Monte Carlo charged particle transport code. Particle fluence area distributions were measured by solid state nuclear track detectors. The design and dosimetric characterisation of the devices are discussed.

Blast colony-forming cell binding from CML bone marrow, or blood, on stromal layers pretreated with G-CSF or SCF.

Blast colony-forming cells (CFU-BL) represent a specific subpopulation of special primitive progenitors characterized by colony formation only in close contact with a preformed stromal layer. CFU-BL derived from bone marrow of chronic myeloid leukaemia (CML) patients have been proved to adhere poorly to bone marrow derived stromal layers suggesting that the appearance of progenitors and precursors in the circulation is due to a defective adhesion of these cells to the bone marrow microenvironment. In the present experiments the effect of short-term incubation of preformed normal bone marrow stroma on the adherence of CML derived CFU-BL was studied. For stroma cultures bone marrow cells were cultured in microplates in the presence of hydrocortisone. Cultures were used when stromal layers became confluent and no sign of haemopoiesis could be observed. CFU-BL were studied by panning plastic non-adherent mononuclear (PNAMNC) bone marrow or blood cells. 8.9 +/- 2.4 colonies/103 PNAMNC (six experiments) were formed from normal bone marrow on stromal layers and 4.8 +/- 2.1 colonies/103 PNAMNC (five experiments) from CML bone marrow. Colony formation from normal bone marrow was not increased if stromal layers were incubated with 100 ng/mL granulocyte colony-stimulating factor (G-CSF) or stem cell factor (SCF). Incubation of stroma with G-CSF or SCF, however, increased the colony formation of PNAMNC from CML bone marrow or blood significantly. These findings suggest that local concentration of haemopoietic growth factors at the time of panning may influence the attachment of CML progenitors to the stroma.

Leukaemogenic potency of WEHI-3B cells grown in vitro or in leukaemic mice.

In order to quantify contaminating leukaemia inducing cells in blood or bone marrow from WEHI-3B bearing BALB/c mice (injected with 10(5) WEHI-3B suspension culture cells), in vitro colony forming leukaemia cells (CFU-L) and survival rate and/or survival time of mice transplanted with cells from WEHI bearing mice or suspension culture were correlated. ED(50) (inoculum inducing leukaemia in 50% of the animals) was 109 CFU-L (33-361) from suspension culture cells. Three weeks after initiation of leukaemia 2 x 10(5) BM or 0.5-2.0 x 10(6) blood cells induced 100% mortality of recipients. After mobilisation with CY or CY and G-CSF, the same amount of blood or BM cells did not induce leukaemia in recipients. A significant negative correlation was found between the survival time of leukaemic mice and the log number of CFU-L inoculated from in vivo sources. In terms of CFU-L cells, leukaemia induction to BM or WBC obtained 3 weeks after leukaemia induction were more potent; those from BM or WBC also obtained at 3 weeks but after mobilisation were less potent inducers than those from suspension culture. These data suggest that CFU-L and leukaemogenic cells are associated, but not identical.

[Surgical trauma in laparoscopic and classical cholecystectomy]. / Operativna trauma pri laparoskopskoj i klasicnoj holecistektomiji.

INTRODUCTION: Accidental or surgical trauma stimulates a response and its intensity is proportional to extent of trauma. The aim of this prospective study was to compare the intensity of the acute-phase reaction and metabolic changes in patients undergoing elective cholecystectomy for chronic cholecystitis. PATIENTS AND METHODS: Sixty patients with cholelithiasis were divided into two groups: thirty patients underwent laparoscopic cholecystectomy (LC group) and thirty patients open cholecystectomy (OC group). Glucose concentration, mean cortisol concentration, C-reactive protein, albumin levels and lactate-dehydrogenase activity were measured preoperatively and postoperatively for up to 48 hours. RESULTS AND DISCUSSION: The examined groups of patients were comparable in regard to age and sex. The duration of operation was similar in both groups. Postoperative hospital stay after laparoscopic operation was significantly shorter than after open cholecystectomy (p < 0.05). The mean glucose concentration (s.e.m.) during the initial 24 hours after surgery was significantly higher (p < 0.05) following open cholecystectomy. The mean cortisol concentration was significantly higher (p < 0.05) following open in regard to laparoscopic operation. Increase in plasma C-reactive protein was significantly higher (p < 0.05) after open cholecystectomy, with maximal levels 48 h after operation. There was a statistically significant decrease (p < 0.05) in albumin concentration after open cholecystectomy. Serum concentration of intracellular enzyme lactate-dehydrogenase increased significantly (p < 0.05) following open in regard to laparoscopic cholecystectomy. CONCLUSION: According to these results, aspects of metabolic and acute-phase responses and tissue damage are reduced following laparoscopic cholecystectomy. The mean postoperative hospital stay was significantly shorter after laparoscopic cholecystectomy with rapid patient recovery.

Secretory function of adrenal cortex in chronic alcoholis.

Three groups of male subjects (healthy subjects, chronic alcoholics with liver cirrhosis and patients with acute viral hepatitis) were included in a 24 hour pattern of excretion of the total and some fractions of 17-ketosteroids (KS), basal concentration of 11-hydroxycorticosteroids (11-OHCS) and dehydroepiandrosterone (DHEA) in plasma, as well as changes of concentration of the same steroids in plasma 15, 30 and 60 minutes after a single i.m. injection of insulin. In regard to healthy subjects and patients with acute viral hepatitis, chronic alcoholics with liver cirrhosis excrete decreased quantities of total and some fractions of 17-KS. In regard to healthy subjects, decreased excretion of the sum androsterone and etiocholanole was established as well as increased DHEA secretion in patients with acute viral hepatitis. In chronic alcoholics with liver cirrhosis basal concentrations of 11-OHCS in plasma and their increase after insulin administration are the same as in healthy subjects, but values of DHEA concentrations in plasma are decreased. It has been pointed to the possibility of damages of the secretory function of adrenal cortex in chronic alcoholics with liver cirrhosis. On the basis of above mentioned results, there is an assumption that adrenal gland primarily provides normal secretion of C21 steroid and thus, satisfying needs for these steroids, increases secretion of DHEA. Follow up of DHEA urinary secretion may provide insight into basal activity of adrenal cortex, whereas the functional state of the liver must be taken into account when interpreting the results.

Lymphoma cell contamination of PBSC: a murine model.

In P-388 bearing BDF1 mice stem cell mobilisation was tested on the survival of lethally irradiated isologous recipients. Contamination of the graft with lymphoma cells was evaluated by the number of clonogenic lymphoma cells (CFU-L) and the ability of the graft to induce lymphoma in non-irradiated recipients. A mobilisation protocol (200 mg/kg cyclophosphamide (CY) i.p. or 200 mg/kg CY followed by 125 microg/kg G-CSF administered every 12 h for 3 consecutive days, starting 14 days after lymphoma initiation) that resulted in a substantial stem cell mobilisation in normal mice, mobilised too few CFU-L to induce lymphoma in the recipients: 50 microl of blood obtained after mobilisation protected lethally irradiated mice but did not induce lymphoma in normal recipients. A minimum graft of bone marrow (2 x 105 cells, with 5580 CFU-L) from untreated P-388 bearing donors killed irradiated recipients presumably by lymphoma and not bone marrow failure. The mobilisation protocol reduced CFU-L so much that no lymphoma-associated death occurred even after the transplantation of 10 x 105 bone marrow cells. These data suggest that, although the PBSC mobilisation protocol may also mobilise some clonogenic lymphoma cells, with the minimum graft size no lymphoma transfer can be observed.

[Diagnostic value of serum erythropoietin levels]. / A szérum eritropoetin szint vizsgálatának diagnosztikai jelentösége.

Erythropoietin (Epo) is a hormone-like glycoprotein regulating erythropoiesis. Under normal conditions Epo stimulates mitoses of the erythroid progenitors and precursors, decreases apoptosis, decreases "ineffective erythropoiesis" and stimulates the synthesis of the specific protein, haemoglobin. Epo producing cells in the kidney sense the O2 tension of kidney tissue and react to hypoxia with increased Epo production and to O2 saturation (polycythaemia) with decreased or completely abolished Epo production. Normal level of Epo in the serum is 3-20 mU/ml. If Epo production is functioning normally there exists a strict inverse correlation between serum Epo level and hematocrit: an exponential increase in Epo level can be observed if hematocrit decreases. Any damage in Epo production lead to inadequate production (e.g. renal anaemias). This paper analyses the Epo content of 278 serum samples assayed in the Laboratory of Experimental Bone Marrow Transplantation of the National Institute of Hematology and Immunology between August 1996 and December 1997 for their diagnostic value. Those samples are primarily discussed where Epo assay was meant to decide diagnosis of polycythaemia vera or those where decision of Epo treatment of anaemic patients depended on their serum Epo level.

Extra dose due to extravehicular activity during the NASA4 mission measured by an on-board TLD system.

A microprocessor-controlled on-board TLD system, 'Pille'96', was used during the NASA4 (1997) mission to monitor the cosmic radiation dose inside the Mir Space Station and to measure the extra dose to two astronauts in the course of their extravehicular activity (EVA). For the EVA dose measurements, CaSO4:Dy bulb dosemeters were located in specially designed pockets of the ORLAN spacesuits. During an EVA lasting 6 h, the dose ratio inside and outside Mir was measured. During the EVA, Mir crossed the South Atlantic Anomaly (SAA) three times. Taking into account the influence of these three crossings the mean EVA/internal dose rate ratio was 3.2. Internal dose mapping using CaSO4:Dy dosemeters gave mean dose rates ranging from 9.3 to 18.3 microGy h-1 at locations where the shielding effect was not the same. Evaluation results of the high temperature region of LiF dosemeters are given to estimate the mean LET.

Doses due to the South Atlantic Anomaly during the Euromir'95 mission measured by an on-board TLD system.

During the Euromir'95 mission, a specially designed microprocessor-controlled thermoluminescent detector (TLD) system, called the 'Pille'95', was used by ESA astronaut Thomas Reiter to measure the cosmic radiation dose inside the Mir space station. One of the experiment's objectives was to determine the dose fraction on Mir due to the South Atlantic Anomaly (SAA) on an orbit inclined at 51.6 degrees and at an altitude of about 400 km. Using an hourly measuring period for 170 h in automatic mode, dose components both of galactic (independent of SAA) and SAA origin were determined. It was found that the maximum dose due to crossing the SAA was equal to 55 microGy. Averaging all the measurements it was calculated that the mean dose rate inside the Mir was 12-14 microGy h-1 and that half of this value was caused by the SAA.

Rapidly mobilizable stem cells. Do they belong to a special subpopulation?

Some characteristics of rapidly mobilized stem cells as a possible distinct subset of the murine bone marrow stem cell population are overviewed. Some of the agents that rapidly mobilize stem cells are toxic and possibly act through disrupting anchorage to the microenvironment. The mobilization occurring days after cytostatics and/or colony-stimulating factors (CSF), however, is a consequence of increased production or differentiation. While stem cells (colony-forming units-spleen; CFU-S) circulating normally in blood have low self-renewal capacity (SRC), SRC of rapidly mobilized CFU-S is closer to that of bone marrow stem cells and is similar to that of the late mobilized stem cells. The survival rate of mice after transplantation of rapidly mobilized stem cells did not differ from that of bone marrow stem cells. One year after transplantation of rapidly mobilized stem cells, the SRC value of bone marrow did not differ from those transplanted with bone marrow cells. Replacement of a rapidly mobilizable stem cell pool requires 48 h under physiological conditions and a longer time after damage to hemopoiesis (irradiation, hydroxyurea injection). Possible physiological mechanisms in the anchorage of stem cells are discussed.

[Diagnostic value of hematopoietic stem cells]. / A vérképzö elödsejtek vizsgálatának diagnosztikai jelentösége.

Proliferation and differentiation of the haemopoietic stem cell are critical for the maintenance of normal haemopoiesis, damage to haemopoiesis, onset of malignant haemopoiesis and repopulation of haemopoiesis after bone marrow transplantation. To evaluate the actual state of haemopoiesis a quantitative assay of haemopoietic stem cells would be needed. Morphology of haemopoietic stem cells cannot be detected by their microscopic morphology. In human practice no stem cell assay is available, therefore, the quantity of human pluripotent stem cells is characterised by the frequency of the progenitors differentiating into erythrocyte, granulocyte or macrophage cell lines (CFU-GEMM, BFU-E, CFU-GM, CFU-Meg). Progenitor cells, can be easily tested by their ability to form in vitro colonies in soft gel cultures in the constant presence of the specific regulator of the cell line ("colony stimulating factor" = CSF). Seven-14 days after explantation of haemopoietic cells colonies containing 40 to 1000 cells are formed. These colonies are proved to be of clonal origin. The quantitative assay of colony forming progenitors has a critical significance in haematological diseases due stem cell damage and bone marrow transplantation. The present paper deals with the diagnostic value of progenitor assays.

Blast colony forming cell-binding capacity of bone marrow stroma from myelodysplastic patients.

A specific stroma function can be quantitatively assessed by counting the stroma-adherent blast cell colonies (CFU-BL) that are formed from normal plastic nonadherent mononuclear bone marrow cells (PNAMNC) after a short-term coincubation ("panning") with the preformed stromal layer. In order to obtain information of stroma function in myelodysplasia (MDS), the "CFU-BL-binding capacity" of stroma from normal bone marrow and from patients with MDS were compared. Stromal cell cultures were established from mononuclear bone marrow cells in microplate cultures cultured with or without 10(-6) M hydrocortisone. CFU-BL-binding capacity was studied by counting blast colonies seven days after panning, and the results were expressed as CFU-BL/10(3) PNAMNC. Normal marrow stromal layers bound CFU-BL only if they were cultured with hydrocortisone, while MDS stromal layers also bound CFU-BL in the absence of hydrocortisone. For further studies of the function of MDS stroma, the effect of growth factors (stem cell factor [SCF], G-CSF, interleukin 3 [IL-3] and their combinations) on CFU-BL binding by normal or MDS stroma has also been compared. Twenty-hour incubation of the stromal layers with a standard dose (100 ng/ml) of various hemopoietic growth factors (IL-3 alone or in combination with SCF, G-CSF alone or in combination with SCF) did not have any effect on CFU-BL binding by normal marrow stroma, but increased the CFU-BL binding by stromal layers from MDS bone marrow. These findings suggest that although stromal microenvironment in MDS is capable of supporting hemopoiesis, bone marrow stroma from MDS patients differs in some characteristics from the normal stroma.

Microprocessor controlled portable TLD system.

An up-to-date microprocessor controlled thermoluminescence dosemeter (TLD) system for environmental and space dose measurements has been developed. The earlier version of the portable TLD system, Pille, was successfully used on Soviet orbital stations as well as on the US Space Shuttle, and for environmental monitoring. The new portable TLD system, Pille'95, consists of a reader and TL bulb dosemeters, and each dosemeter is provided with an EEPROM chip for automatic identification. The glow curve data are digitised and analysed by the program of the reader. The measured data and the identification number appear on the LED display of the reader. Up to several thousand measured data together with the glow curves can be stored on a removable flash memory card. The whole system is supplied either from built-in rechargeable batteries or from the mains of the space station.

[Polycytemia vera in an 11-year old girl]. / Valódi polycythaemia 11 éves leányban.

The case of an 11 year old girl with three line type of polycythaemia vera, with 4 cm splenomegaly and a plethoric complexion is presented. Peripheral blood values were as follows: RBC: 7.75 x 10(12)/l, Hb: 18.8 g/l, WBC: 15.2 x 10(9)/l, platelets: 920 x 10(9)/l. Serum erythropoietin level: < 1 mU/ml. In vitro, erythroid colonies developed from the bone marrow in the absence of added erythropoetin. For three years the haematocrit value has been controlled by regular venesections. Since extreme thrombocytosis developed, the treatment was continued with interferon alpha. Different treatment protocols are discussed.