Assessment of diagnostic utility of multivariate diagnostic models in differential diagnosis of ovarian tumors

Introduction: Ovarian cancer (OC) diagnosis remains a clinical challenge due to lack of early symptoms and insufficient accuracy of the available diagnostic methods. The purpose of this study was to determine whether osteopontin could be useful in differential diagnosis of ovarian tumors.Material and methods: Serum samples from 92 patients qualified for surgical treatment due to ovarian mass were divided into 2 groups according to the histopathological result: OC including borderline ovarian tumors (n = 39) and benign ovarian tumors (BOTs) (n = 53). CA125, HE4 and osteopontin concentrations were measured in all patients. Areas under the receiver operating characteristic curves (AUC of ROC) were used to compare the discriminative ability of the univariate and multivariate diagnostic models.Results: The addition of osteopontin to ROMA significantly improved the diagnostic performance of the test in 3 of the 5 analyses: 1) in the OC vs BOT group (from AUC of 0.955 to 0.975), 2) in premenopausal women OC vs BOT (from AUC of 0.828 to 0.892) and 3) in the FIGO I-II stage OC vs BOT (from AUC of 0.865 to 0.895). It did not alter the diagnostic performance of multifactor tests in the group of postmenopausal women nor in OC FIGO III-IV stage group. Osteopontin was also thebest single marker to differentiate between early stage OC and BOTs (AUC of 0.863).Conclusions: Osteopontin improves the diagnostic performance of a multimarker OC diagnostic test and could be useful in differential diagnosis of ovarian tumors, especially in pre-menopausal women and for early stage OC

Metabolomika i proteomika w diagnostyce nieswoistych chorób zapalnych jelit

Ulcerative colitis and Crohn’s disease belong to the group of inflammatory bowel diseases and are characterized by chronic inflammation of gastrointestinal tract. The pathogenesis of these conditions is multifactorial and not fully understood. The diagnostic process of inflammatory bowel diseases is often time consuming and involves many different diagnostic tests. Researchers are looking for novel diagnostic tools, that would lead to early and correct diagnosis, optimal treatment and would be possibly least invasive. Novel technolog ies, such as metabolomics and proteomics show great promise for future clinical use. Meta bolomics involves analysis of metabolites in cells and tissues, whereas proteomics is based on the analysis protein and peptide profile, that are changing depending on underlying disease.Wrzodziejące zapalenie jelita grubego i choroba Leśniowskiego-Crohna to przewlekłe choroby zapalne przewodu pokarmowego, których patogeneza jest wieloczynnikowa i nie do końca zbadana. Proces diagnostyczny prowadzący do właściwego leczenia jest często czasochłonny. Od lat trwają badania zmierzające do odkrycia wiarygodnych i minimalnie inwazyjnych testów diagnostycznych, służących do szybkiego rozpoznawania tych chor ób i poprawy opieki nad pacjentami. Duże nadzieje niosą ze sobą nowe dziedziny nauki, takie jak metabolomika i proteomika, które mogą prowadzić do odkr ycia nowych biomarkerów tych schor zeń. Zajmują się oceną metabolitów (metabolomika) oraz białek i peptydów (pr oteomika) w komórkach i tkankach, których skład zmienia się w zależności od stanu chorobowego

New CZE-DAD method for honeybee venom analysis and standardization of the product

The aim of this study was to develop a new precise and accurate CZE-DAD method for honeybee venom analysis using cytochrome c as an internal standard. The 64.5 cm total length, 56 cm effective length, 75 μm ID, and 360 μm OD uncoated fused-silica capillary was used. The samples were injected into the capillary under a 50-mbar pressure for 7 s. There were 15 kV of electric field across the capillary applied. The current intensity was 26 μA. The separation was carried out at 25 °C. The analysis was run with the normal electrode polarity. The following steps and parameters were taken into account for the validation of the developed method: selectivity, precision, accuracy, linearity, limit of detection and limit of quantitation. All steps of the validation procedure proved that the developed analytical procedure was suitable for its intended purpose. Possibly this was the first study in which several honeybee venom components were separated and five of them were identified by capillary zone electrophoresis. In addition, the developed method was applied for quantitative analysis of 38 honeybee venom samples. The content (relative to the dry venom mass) of analyzed peptides in honeybee venom samples collected in 2002–2007 was as follows: apamine from 0.93% to 4.34% (mean, 2.85 ± 0.79%); mast cell degranulating peptide (MCDP) from 1.46% to 4.37% (mean, 2.82 ± 0.64%); phospholipase A2 from 7.41% to 20.25% (mean, 12.95 ± 3.09%); melittin from 25.40% to 60.27%, (mean, 45.91 ± 9.78%). The results were compared with the experimental data obtained for the same venom samples analyzed earlier by the HPLC method. It was stated that HPCE and HPLC data did not differ significantly and that the HPCE method was the alternative for the HPLC method. Moreover, using the results obtained principal component analysis (PCA) was applied to clarify the general distribution patterns or similarities of four major honeybee venom constituents collected from two different bee strains in various months and years. PCA has shown that the strain of bee appears to be the only criteria for bee venom sample classification. Strong correlations between apamine, MCDP, phospholipase A2, and melittin were confirmed. These correlations have to be taken into account in the honeybee venom standardization. The developed method due to its simplicity can be easily automated and incorporated into routine operations both in the bee venom identification, quality control, and standardization of the product

Standard methods for Apis mellifera venom research

Honey bees have a sting which allows them to inject venomous substances into the body of an opponent or attacker. As the sting originates from a modified ovipositor, it only occurs in the female insect, and this is a defining feature of the bee species that belong to a subclade of the Hymenoptera called Aculeata. There is considerable interest in bee venom research, primarily because of an important subset of the human population who will develop a sometimes life threatening allergic response after a bee sting. However, the use of honey bee venom goes much further, with alleged healing properties in ancient therapies and recent research. The present paper aims to standardize selected methods for honey bee venom research. It covers different methods of venom collection, characterization and storage. Much attention was also addressed to the determination of the biological activity of the venom and its use in the context of biomedical research, more specifically venom allergy. Finally, the procedure for the assignment of new venom allergens has been presented. Las abejas meliferas tienen un aguijon que les permite inyectar sustancias venenosas en el cuerpo de un oponente o atacante. El aguijon es un ovipositor modificado que solo se manifiesta en el insecto hembra, siendo este una caracteristica que define a las especies de abejas que pertenecen al subclado de himenopteros llamada Aculeata. Hay un interes considerable en la investigacion del veneno de abeja, principalmente debido a que un porcentaje importante de la poblacion humana desarrollara una respuesta alergica - a veces mortal - a la picadura de abeja. Sin embargo, el uso del veneno de la abeja melifera abarca mucho mas, con presuntas propiedades curativas en terapias antiguas e investigaciones recientes. El presente trabajo tiene como objetivo estandarizar metodos seleccionados para la investigacion del veneno de las abejas meliferas. Cubre diferentes metodos de recoleccion, caracterizacion y almacenamiento de veneno. Tambien se presto mucha atencion a la determinacion de la actividad biologica del veneno y su uso en el contexto de la investigacion biomedica, mas especificamente la alergia al veneno. Finalmente, se ha presentado el procedimiento para la asignacion de nuevos alergenos de veneno

Assessing circadian rhythms in propofol PK and PD during prolonged infusion in ICU patients

This study evaluates possible circadian rhythms during prolonged propofol infusion in patients in the intensive care unit. Eleven patients were sedated with a constant propofol infusion. The blood samples for the propofol assay were collected every hour during the second day, the third day, and after the termination of the propofol infusion. Values of electroencephalographic bispectral index (BIS), arterial blood pressure, heart rate, blood oxygen saturation and body temperature were recorded every hour at the blood collection time points. A two-compartment model was used to describe propofol pharmacokinetics. Typical values of the central and peripheral volume of distribution and inter-compartmental clearance were VC = 27.7 l, VT = 801 l, and CLD = 2.73 l/min. The systolic blood pressure (SBP) was found to influence the propofol metabolic clearance according to Cl (l/min) = 2.65·(1 − 0.00714·(SBP − 135)). There was no significant circadian rhythm detected with respect to propofol pharmacokinetics. The BIS score was assessed as a direct effect model with EC50 equal 1.98 mg/l. There was no significant circadian rhythm detected within the BIS scores. We concluded that the light–dark cycle did not influence propofol pharmacokinetics and pharmacodynamics in intensive care units patients. The lack of night–day differences was also noted for systolic blood pressure, diastolic blood pressure and blood oxygenation. Circadian rhythms were detected for heart rate and body temperature, however they were severely disturbed from the pattern of healthy patients

Bee venom allergy in beekeepers

Beekeepers are a particular group of people highly exposed to stings and allergy to bee venom due to the nature of their work. Positive skin test results or bee venom specific IgE are observed in up to 60% of beekeepers. Reaction to the bee sting occurs most often as a normal local reaction, but a person allergic to bee venom may experience large local reaction, systemic symptoms, including anaphylactic shock. In patients with confirmed systemic allergic reaction in the past, basic diagnostic tests such as skin tests and bee venom specific IgE must be performed. Where appropriate, additional tests such as component- -resolved diagnosis, cell-based assays, tryptase or the venom-specific IgG4 levels should be conducted. Studies have shown significantly higher tryptase and specific IgG4 levels in beekeepers. Moreover, a correlation between clinical symptoms after the sting and the phospholipase A2 specific IgE concentration was observed. During venom-specific immunotherapy, increasing the dose to 200 µg should be considered in beekeepers.Pszczelarze są szczególną grupą osób najbardziej narażonych na użą- dlenia i alergię na jad pszczeli z racji wykonywanej pracy. Dodatnie wyniki testów skórnych lub obecność swoistych dla jadu pszczelego IgE obserwowane są nawet u 60% badanych pszczelarzy. Reakcja po użądleniu przez pszczołę najczęściej występuje pod postacią normalnej reakcji miejscowej, ale u osoby uczulonej na jad pszczeli może wystą- pić nasilona reakcja miejscowa lub objawy ogólne, do wstrząsu anafilaktycznego włącznie. U pacjentów z reakcją systemową w wywiadzie należy wykonać podstawowe badania diagnostyczne: testy skórne i oznaczenia swoistych dla jadu sIgE, a w uzasadnionych przypadkach badania dodatkowe, takie jak: oznaczenia sIgE dla poszczególnych alergenów jadu pszczelego (diagnostyka komponentowa), testy komórkowe, oznaczanie poziomu tryptazy czy stężenia swoistych dla jadu IgG4. Badania przeprowadzone u pszczelarzy wykazały istotnie wyższe stę- żenia tryptazy oraz swoistych IgG4 w tej grupie zawodowej. Ponadto zaobserwowano korelację nasilenia objawów klinicznych po użądleniu ze stężeniem sIgE dla fosfolipazy A2 . Podejmując decyzję o prowadzeniu swoistej immunterapii jadem pszczelim u pszczelarza należy rozważyć zwiększenie dawki podtrzymującej do 200 µg

Application of Metabolomic Tools for Studying Low Molecular-Weight Fraction of Animal Venoms and Poisons

Both venoms and poisonous secretions are complex mixtures that assist in defense, predation, communication, and competition in the animal world. They consist of variable bioactive molecules, such as proteins, peptides, salts and also metabolites. Metabolomics opens up new perspectives for the study of venoms and poisons as it gives an opportunity to investigate their previously unexplored low molecular-weight components. The aim of this article is to summarize the available literature where metabolomic technologies were used for examining the composition of animal venoms and poisons. The paper discusses only the low molecular-weight components of venoms and poisons collected from snakes, spiders, scorpions, toads, frogs, and ants. An overview is given of the analytical strategies used in the analysis of the metabolic content of the samples. We paid special attention to the classes of compounds identified in various venoms and poisons and potential applications of the small molecules (especially bufadienolides) discovered. The issues that should be more effectively addressed in the studies of animal venoms and poisons include challenges related to sample collection and preparation, species-related chemical diversity of compounds building the metabolome and a need of an online database that would enhance identification of small molecule components of these secretions

Mass spectrometry-based proteomics techniques and their application in ovarian cancer research

Abstract Ovarian cancer has emerged as one of the leading cause of gynecological malignancies. So far, the measurement of CA125 and HE4 concentrations in blood and transvaginal ultrasound examination are essential ovarian cancer diagnostic methods. However, their sensitivity and specificity are still not sufficient to detect disease at the early stage. Moreover, applied treatment may appear to be ineffective due to drug-resistance. Because of a high mortality rate of ovarian cancer, there is a pressing need to develop innovative strategies leading to a full understanding of complicated molecular pathways related to cancerogenesis. Recent studies have shown the great potential of clinical proteomics in the characterization of many diseases, including ovarian cancer. Therefore, in this review, we summarized achievements of proteomics in ovarian cancer management. Since the development of mass spectrometry has caused a breakthrough in systems biology, we decided to focus on studies based on this technique. According to PubMed engine, in the years 2008–2010 the number of studies concerning OC proteomics was increasing, and since 2010 it has reached a plateau. Proteomics as a rapidly evolving branch of science may be essential in novel biomarkers discovery, therapy decisions, progression predication, monitoring of drug response or resistance. Despite the fact that proteomics has many to offer, we also discussed some limitations occur in ovarian cancer studies. Main difficulties concern both complexity and heterogeneity of ovarian cancer and drawbacks of the mass spectrometry strategies. This review summarizes challenges, capabilities, and promises of the mass spectrometry-based proteomics techniques in ovarian cancer management