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

Wstępne wyniki pierwszego w Polsce programu przesiewowego w tętniaku aorty brzusznej w województwie kujawsko-pomorskim

Background. Abdominal aortic aneurysm (AAA) is called a widening of the aorta below the renal arteries with a diameter more than 3 cm. The prevalence of AAA is estimated at 4–8% in men aged 65 years or more. The mortality rate for patients with ruptured AAA is 65%, while the mortality rate for aortic aneurysm repair surgery in elective patients undergoing pre-screened is 3%. The aim of this study was to determine the prevalence of AAA among men aged 60 years and older undergoing targeted ultrasonography of abdominal aorta. The additional aim was to determine the relationship between specific risk factors and the AAA, and to determine the prevalence of AAA in patients subgroups specified by the criteria for the screening programmes for AAA. Material and methods. Based on available data from the literature, study populations was defined as men aged 60 years and older. Applied research methods were as follow: 1) the questionnaire; 2) measurement of physical characteristics of aorta (diameter of abdominal aorta by ultrasound scan). The questionnaire consisted of two parts: 1) identification and demographic data; 2) the presence of risk factors. Analysis of data from the questionnaire determined the relationship between the risk factors and the AAA, whereas ultrasonography of the aorta, detected the disease (abdominal aortic aneurysm) and its prevelance. Statistical analysis of the data was performed using the software STATISTICA 9 (StatSoft, Inc., 2009). Results. Based on 1556 ultrasound scans, the abdominal aortic aneurysm was diagnosed in 94 men aged 60 years and older. The prevalence of AAA in the study group was 6.0%. Risk factors associated with the AAA were analyzed in two groups: I — AAA (n = 94) and II — normal aorta (n = 1464). The three risk factors were significantly associated with the AAA: age, smoking and family history, whereas the incidence of AAA in males at 65 years old was 4.0%, and a group of men between 65 and 75 years of age, smoking cigarettes ever, was 7.4%. Conclusions. In the obtained data the prevalence of AAA was similar to that of literature. Additionally, the positive relationship between age, smoking, and AAA was detected. Acta Angiol 2012; 18, 1: 9–17Wstęp. Tętniakiem aorty brzusznej (TAB) określa się poszerzenie aorty poniżej odejścia tętnic nerkowych o średnicy powyżej 3 cm. Częstość występowania TAB szacuje się na 4–8% u mężczyzn w wieku 65 lat i więcej. Współczynnik umieralności dla pacjentów z pękniętym TAB wynosi 65%, natomiast śmiertelność pooperacyjna dla operacji naprawczych tętniaka aorty w trybie planowym wśród pacjentów poddanych wcześniej badaniu przesiewowemu wynosi 3%. Podstawowym celem pracy było określenie częstości występowania (chorobowość, prevalence) TAB wśród mężczyzn w wieku 60 lat i starszych poddanych ukierunkowanemu badaniu ultrasonograficznemu aorty w odcinku brzusznym. Dodatkowo zbadano zależności między określonymi czynnikami ryzyka a TAB oraz częstość występowania TAB w subgrupach pacjentów określonych według kryteriów programów badań przesiewowych w kierunku TAB. Materiał i metody. Na podstawie dostępnego piśmiennictwa zdefiniowano populację narażoną, tj. mężczyzn w wieku 60 lat i starszych. Zastosowano następujące metody badawcze: 1) metodę kwestionariuszową; 2) pomiar cechy fizycznej (badanie ultrasonograficzne średnicy aorty w odcinku brzusznym). Kwestionariusz składał się z dwóch części: 1) dotyczącej danych identyfikacyjnych i demograficznych; 2) dotyczącej występowania czynników ryzyka. Analiza danych z kwestionariusza umożliwiła określenie zależności między badanymi czynnikami ryzyka a TAB, natomiast poddanie zdefiniowanej części populacji badaniu ultrasonograficznemu aorty umożliwiło wykrycie choroby (tętniak aorty brzusznej) i określenie częstość jej występowania. Analizę statystyczną uzyskanych danych przeprowadzono z wykorzystaniem oprogramowania STATISTICA 9 (StatSoft, Inc. 2009). Wyniki. Na podstawie wykonanych 1556 badań ultrasonograficznych aorty brzusznej tętniak zdiagnozowano u 94 mężczyzn w wieku 60 lat i starszych. Częstość występowania (prevalence) TAB w badanej grupie wyniosła 6,0%. Czynniki ryzyka związane z TAB analizowano, wyodrębniając dwie grupy: I — TAB (n = 94) i II — aorta prawidłowa (n = 1464). Istotny statystycznie związek z TAB miały 3 czynniki ryzyka: wiek, palenie tytoniu i występowanie rodzinne TAB. Częstość występowania TAB w grupie mężczyzn w 65. roku życia wyniosła 4,0%, natomiast w grupie mężczyzn między 65. a 75. rokiem życia, palących kiedykolwiek tytoń, wyniosła 7,4%. Wnioski. Na podstawie uzyskanych danych stwierdzono częstość występowania TAB podobną do tej z piśmiennictwa. Dodatkowo stwierdzono także zależności między wiekiem, paleniem tytoniu i występowaniem rodzinnym TAB a zachorowaniem na TAB. Acta Angiol 2012; 18, 1: 9–1

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

The correlation between anti phospholipase A 2 specific IgE and clinical symptoms after a bee sting in beekeepers

Introduction: Beekeepers are a group of people with high exposure to honeybee stings and with a very high risk of allergy to bee venom. Therefore, they are a proper population to study the correlations between clinical symptoms and results of diagnostic tests. Aim: The primary aim of our study was to assess the correlations between total IgE, venom- and phospholipase A 2 -specific IgE and clinical symptoms after a bee sting in beekeepers. The secondary aim was to compare the results of diagnostic tests in beekeepers and in individuals with standard exposure to bees. Material and methods: Fifty-four individuals were divided into two groups: beekeepers and control group. The levels of total IgE (tIgE), venom-specific IgE (venom sIgE), and phospholipase A 2 -specific IgE (phospholipase A 2 sIgE) were analyzed. Results: Our study showed no statistically significant correlation between the clinical symptoms after a sting and tIgE in the entire analyzed group. There was also no correlation between venom sIgE level and clinical symptoms either in beekeepers or in the group with standard exposure to bees. We observed a statistically significant correlation between phospholipase A 2 sIgE level and clinical signs after a sting in the group of beekeepers, whereas no such correlation was detected in the control group. Significantly higher venom-specific IgE levels in the beekeepers, as compared to control individuals were shown. Conclusions : In beekeepers, the severity of clinical symptoms after a bee sting correlated better with phospholipase A 2 sIgE than with venom sIgE levels

Immune and clinical response to honeybee venom in beekeepers

OBJECTIVE The aim of the study was to assess immune response to honeybee venom in relation to the degree of exposure, time after a sting and clinical symptoms. Material and Methods Fifty-four volunteers were divided into 2 groups: beekeepers and a control group. The serum levels of total IgE (tIgE), bee venom-specific IgE (venom sIgE), phospholipase A 2 -specific IgE (phospholipase A 2 sIgE), tryptase and venom-specific IgG4 (venom sIgG4) were determined. In beekeepers, diagnostic tests were performed within 3 hours following a sting and were repeated after a minimum of 6 weeks from the last sting. In individuals from the control group, the tests were performed only once, without a sting. Results The tests showed significant differences in venom sIgE (beekeepers' median = 0.34 kUA/l, control group median = 0.29 kUA/l), baseline serum tryptase (beekeepers' median = 4.25 µg/l, control group median = 2.74 µg/l) and sIgG4 (beekeepers' median = 21.2 mgA/l, control group median = 0.14 mgA/l), confirming higher levels of the tested substances in the beekeepers than in the control group. A significant positive correlation was observed between phospholipase A 2 sIgE concentration and severity of clinical symptoms after a sting in the group of beekeepers. It was also demonstrated that the clinical symptoms after a sting became less severe with increasing age of the beekeepers. Conclusions The differences in the immune response to a bee sting between the beekeepers and individuals not exposed to bees were probably due to the high exposure of the beekeepers to honeybee venom allergens. This may suggest a different approach to the bee venom allergy diagnostic tests in this occupational group

Immune and clinical response to honeybee venom in beekeepers

OBJECTIVE The aim of the study was to assess immune response to honeybee venom in relation to the degree of exposure, time after a sting and clinical symptoms. Material and Methods Fifty-four volunteers were divided into 2 groups: beekeepers and a control group. The serum levels of total IgE (tIgE), bee venom-specific IgE (venom sIgE), phospholipase A 2 -specific IgE (phospholipase A 2 sIgE), tryptase and venom-specific IgG4 (venom sIgG4) were determined. In beekeepers, diagnostic tests were performed within 3 hours following a sting and were repeated after a minimum of 6 weeks from the last sting. In individuals from the control group, the tests were performed only once, without a sting. Results The tests showed significant differences in venom sIgE (beekeepers' median = 0.34 kUA/l, control group median = 0.29 kUA/l), baseline serum tryptase (beekeepers' median = 4.25 µg/l, control group median = 2.74 µg/l) and sIgG4 (beekeepers' median = 21.2 mgA/l, control group median = 0.14 mgA/l), confirming higher levels of the tested substances in the beekeepers than in the control group. A significant positive correlation was observed between phospholipase A 2 sIgE concentration and severity of clinical symptoms after a sting in the group of beekeepers. It was also demonstrated that the clinical symptoms after a sting became less severe with increasing age of the beekeepers. Conclusions The differences in the immune response to a bee sting between the beekeepers and individuals not exposed to bees were probably due to the high exposure of the beekeepers to honeybee venom allergens. This may suggest a different approach to the bee venom allergy diagnostic tests in this occupational group

Ocena wskaźnika kostka–ramię i ryzyka chorób sercowo-naczyniowych w populacji kobiet po menopauzie

Background: The incidence of peripheral artery disease (PAD) and cardiovascular (CV) events in the female population has been on the increase. Aim: To analyse the risk factors of a CV event and PAD in women and to assess the usefulness of the ankle–brachial index (ABI). Methods: Evaluation of selected parameters in a cohort of 365 women living in the same district. The following data were prospectively recorded: weight, height, waist size, hip circumference, smoking, the intima–media complex, ABI value, and laboratory results. PAD symptoms, CV events and neurological events were noted. ABI was analysed assuming pathology for values: ≤ 0.9 or ≤ 1.0. Results: Age, plasma glucose level, atrial fibrillation, and nicotine addiction were correlated independently with CV disease and stroke (p < 0.001). The high-density lipoprotein cholesterol level, height, and systolic blood pressure were correlated independently with ABI values (p < 0.05). There was no correlation between the occurrence of a CV event in the past and the ABI, irrespective of the cut-off point for the reference value (p = NS). Conclusions: There is no evidence that stricter criteria for the assessment of ABI better represent the vascular status in the female population.Wstęp: Choroba tętnic obwodowych (PAD) coraz częściej dotyczy populacji kobiet. Odmienności w symptomatologii i prze­biegu klinicznym sugerują, że konieczna jest weryfikacja zaleceń diagnostycznych. Cel: Analiza czynników ryzyka chorób sercowo-naczyniowych i PAD u kobiet oraz ocena przydatności wskaźnika kostka–ramię (ABI). Metody: Analizowano wybrane parametry w populacji 365 kobiet zamieszkujących obszar jednego powiatu. Prospektywnie rejestrowano następujące parametry: masa ciała, wzrost, obwód w talii, obwód bioder, palenie tytoniu, kompleks intima–media, wartość ABI oraz wybrane parametry laboratoryjne. Odnotowywano, czy u pacjentów występowały objawy choroby tętnic obwodowych, chorób sercowo-naczyniowych oraz incydenty neurologiczne. Wskaźnik kostka–ramię analizowano przy założeniu patologii dla wartości ≤ 0,9 oraz ≤ 1,0. Zebrane parametry poddano analizie statystycznej. Wyniki: Wiek, stężenie glukozy, migotanie przedsionków oraz nikotynizm były niezależnymi predykatorami chorób sercowo-naczyniowych i mózgowych (p < 0,001). Stężenie cholesterolu frakcji lipoprotein o wysokiej gęstości, wartość skurczowego ciśnienia tętniczego i wzrost były niezależnymi czynnikami wpływającymi na ABI (p < 0,05). Nie wykazano zależności między ABI oraz incydentami sercowo-naczyniowymi w przeszłości, niezależnie od przyjętej wartości progowej (p = NS). Wnioski: Nie wykazano, że przyjęcie wartości ABI ≤ 1,0 za patologiczną lepiej korelowało z występowaniem chorób sercowo-naczyniowych w badanej populacji kobiet