Elektroforeza serumskih beljakovin je metoda ločevanja beljakovin na podlagi njihovih lastnosti. Pod vplivom električne napetosti se serumske beljakovine razvrstijo v šest frakcij, med katerimi ...predstavljajo albumini največjo frakcijo, ostale frakcije pa pet vrst globulinov. Elektroforeza serumskih beljakovin ima v prvi vrsti klinični pomen pri ugotavljanju diseminiranega plazmocitoma in drugih bolezni serumskih beljakovin, sicer pa lahko z elektroforezo prepoznamo morfološke vzorce glede na akutno ali kronično vnetje, različne malignome, bolezni jeter in ledvic.
Background: Identification, quantification and typing of MProteins (MP) play an important role in the diagnosis, classification and monitoring of monoclonal gammopathies both of malignant origin (eg. ...Multiple Myeloma) and of unknown origin. Previous evidence attests that MGUS (Monoclonal Gammopathy of Undetermined Significance) detected by agarose gel electrophoresis has a prevalence of 3.2% in the general population. Therefore, our study aimed to verify this data by means of capillary zone electrophoresis (CZE).
Methods: CZE was performed to evaluate the prevalence of M-Protein (MP) in 44.474 consecutive outpatients of all ages with a prescription for serum protein electrophoresis over a 2-year period (2008 and 2009). All MPs that were identified were then typed by immunofixation electro pho - resis on agarose gel (IFE).
Results: In subjects aged over 50 (23.408, i.e., 52.6% of the whole cohort) MP ≤30 g/L (MGUS) was identified in 6.0% of cases, with a frequency nearly double than that previously reported. The population was then divided into ten-year age groups: the 71-80 age group had the highest percentage of MP (29%), followed by 61-70 (27%), 51-60 (18%), 81-90 (12%), 41-50 (8%), 31-40 (3%), >90 (2%) and <30 (1%). The frequency of MP types (IFE) was the same in each age group, with IgG Kappa being the most represented class.
Conclusions: According to the high MGUS prevalence observed in this study, these results may be useful especially for general practitioners, because the identification even of small MP (analytical sensitivity: 0.5 g/L) may help optimize clinical management.
Uvod: Identifikacija, kvantifikacija i tipizacija M-proteina (MP) imaju važnu ulogu u dijagnostikovanju, klasifikovanju i praćenju monoklonskih gamapatija kako malignog (npr. multipli mijelom) tako i nepoznatog porekla. Prethodni dokazi pokazuju da monoklonska gamapatija neodređenog značaja (MGNZ) koja se otkriva elektroforezom na agaroznom gelu ima prevalenciju od 3,2% u opštoj (populaciji. Stoga je cilj ove studije bio da verifikuje navedene podatke putem elektroforeze kapilarne zone.
Metode: Kapilarna elektroforeza je izvršena da bi se utvrdi- la prevalencija M-proteina (MP) kod 44.474 uzastopnih kliničkih pacijenata svih uzrasta kod kojih je u toku dve godine (2008. i 2009) prepisana elektroforeza proteina u serumu. Identifikovani su svi MP a zatim tipizirani imunofiksacionom elektroforezom na agaroznom gelu.
Rezultati: Kod ispitanika sfarijih od 50 godina (23.408, tj. 52,6% ukupnog broja) MP <30 g/L (MGNZ) idenfifikovan je u 6,0% slučajeva, sa gotovo dva puta većom učestalošću nego što je prethodno procenjeno. Populacija je zatim podeljena na starosne grupe raspona po deset godina: grupa 71-80 imala je najved procenat MP (29%), a slede grupe 61-70 (27%), 51-60 (18%), 81-90 (12%), 41-50 (8%), 31-40 (3%), > 90 (2%) i < 30 (1%). Učestalost tipova MP (imunofiksaciona elektroforeza na agaroznom gelu) bila je isfa u svim starosnim grupama, dok je najzastupljenija klasa bila IgG Kappa.
Zakljiučak: Na osnovu velike uo~ene prevalencije MGNZ, zaključujemo da ovi rezultati mogu biti naročito korisni za lekare op{te prakse, po{to identifikacija čak i malih MP (analitička osetljivost: 0,5 g/L) može doprineti optimizaciji kliničkog menad`menta.
Proteomics of heavy metal toxicity in plants Cvjetko, Petra; Zovko, Mira; Balen, Biljana
Arhiv za higijenu rada i toksikologiju,
03/2014, Letnik:
65, Številka:
1
Journal Article
Recenzirano
Odprti dostop
Plants endure a variety of abiotic and biotic stresses, all of which cause major limitations to production. Among abiotic stressors, heavy metal contamination represents a global environmental ...problem endangering humans, animals, and plants. Exposure to heavy metals has been documented to induce changes in the expression of plant proteins. Proteins are macromolecules directly responsible for most biological processes in a living cell, while protein function is directly influenced by posttranslational modifications, which cannot be identified through genome studies. Therefore, it is necessary to conduct proteomic studies, which enable the elucidation of the presence and role of proteins under specific environmental conditions. This review attempts to present current knowledge on proteomic techniques developed with an aim to detect the response of plant to heavy metal stress. Significant contributions to a better understanding of the complex mechanisms of plant acclimation to metal stress are also discussed.
Biljke su neprestano izložene različitim čimbenicima abiotičkog i biotičkog stresa koji nepovoljno utječu na njihovu produktivnost. Teški metali kao česti zagađivači okoliša vrlo nepovoljno djeluju na sva živa bića, uključujući biljke, životinje i ljude. Poznato je da teški metali mogu mijenjati ekspresiju biljnih proteina. Proteine ubrajamo u biološki vrlo važne makromolekule čija je aktivnost u stanici izravno ovisna o posttranslacijskim modifikacijama, koje nije moguće pratiti na razini genoma. Stoga je nužno provoditi proteomska istraživanja kako bi se razotkrila prisutnost i uloga proteina u različitim vrstama okolišnog stresa. U ovom radu sažete su različite tehnike i metode istraživanja učinaka teških metala na biljni proteom, uključujući i sažet osvrt na složene mehanizme odgovora biljke na stres izazvan teškim metalima.
Cilj: Vankomicin je antibiotik koji se koristi za liječenje infekcija uzrokovanih Gram- -pozitivnim bakterijama, no kod nekih pacijenata uzrokuje ozljedu bubrega. Budući da standardni biomarkeri ...ozljede bubrega ne pokazuju dovoljnu osjetljivost i specifičnost, potrebno je pronaći nove biomarkere kako bismo mogli ranije detektirati poremećaje bubrežne funkcije te pravovremenim prekidom ili promjenom terapije spriječiti daljnje ozljede bubrega. Cilj je rada bio usporediti proteomske profile urina dviju pacijentica oboljelih od upale pluća koje su primale terapiju vankomicinom sa i bez kliničkih znakova ozljede bubrega uzrokovanih navedenim antibiotikom, kako bismo identificirali nove potencijalne proteinske biomarkere akutne ozljede bubrega uzrokovanog vankomicinom. Prikaz slučaja: Uzorci urina prikupljeni su treći i sedmi dan terapije vankomicinom od pacijentice sa i pacijentice bez kliničkih znakova bubrežne disfunkcije te su analizirani dvodimenzionalnom gel elektroforezom kombiniranom s MALDI-TOF/TOF masenom spektrometrijom. Vidljive razlike u proteomskim profilima urina između dviju pacijentica javljaju se treći, a naročito su izražene sedmi dan terapije. Dvodimenzionalna gel elektroforeza kombinirana s masenom spektrometrijom potvrdila je već ranije dokazanu ulogu proteina neutrofilnog lipokalina povezanog s gelatinazom (NGAL, engl. neutrophil gelatinase-associated lipocalin) kao biomarkera akutne ozljede bubrega, te otkrila ekspresiju citoskeletnog keratina 1 tipa II i retinol-vezujućeg proteina 4 (RBP4, engl. retinol-binding protein 4) samo u urinu pacijentice kod koje je terapija vankomicinom uzrokovala poremećaj bubrežne funkcije. Zaključci: Citoskeletni keratin 1 tipa II i retinol-vezujući protein 4 mogli bi predstavljati nove potencijalne biomarkere ozljede bubrega uzrokovane vankomicinom. Potrebne su daljnje studije na većem broju pacijenata kako bi se validirali dobiveni rezultati i utvrdio njihov potencijalni dijagnostički i klinički značaj.
Abstract Biodiversity is a key concept in finding important features of new microorganisms. Microorganisms play an important role in the soil ecosystem and participate, among others, in such ...processes as the maintenance of soil structure, humification, release of organic compounds, disposal of pollutants and transformation of organic matter. The maintenance of competent state of soil microbial communities, i.e. the appropriate microorganism count, activity and diversity, is a necessary condition for the functioning of a highly complex system such as the soil. Phyllosphere bacteria have the potential to influence plant biogeography and ecosystem function through their influence on plant performance under different environmental conditions, but the drivers of variation in leaf-associated bacterial biodiversity among host plants are not well understood. Hence, undoubtedly, an important research aspect is the selection and development of indicators to evaluate microbial biodiversity of the soil and plant phyllosphere. In this publication, selected molecular methods used for the diversity assessment of microorganisms have been presented. 1. Introduction. 2. Denaturing Gradient Gel Electrophoresis DGGE, Temperature Gradient Gel Electrophoresis TGGE, 3. SSCP – single strand conformation polymorphism. 4. Real-Time Quantitative PCR. 5. Summary
The identification of the new form of hemoglobin in fetus red blood cells (HbF), different from adult hemoglobin (HbA), was made over one hundred years ago. Since this time, various methods of fetal ...hemoglobin measurement have been designed. Most of them are based on the different biochemical characteristics of HbF. Fetal hemoglobin's affinity for oxygen is substantially greater than that of adult hemoglobin. The first techniques for the determination of fetal hemoglobin were based on its resistance to denaturation by alkaline solutions. Currently, the measurement of hemoglobin F is an important part of diagnosis of sickle cell disease, thalassemia, hereditary persistence of fetal hemoglobin, and fetomaternal hemorrhage. Amongst the most commonly used and clinically important methods, high-performance liquid chromatography, capillary electrophoresis, isoelectric focusing, capillary isoelectric focusing, Kleihauer–Betke test, and flow cytometry should be listed.
Agarose gel electrophoresis is a well established technique routinely used in clinical laboratories for screening protein abnormalities in various biological fluids (serum, urine, CSF). It is based ...on the principles of zone electrophoresis. Electrophoretograms are evaluated visually for the presence of quantitatively or qualitatively abnormal protein bands. The technique is used for electrophoresis of serum, urine, CSF proteins, enzymes (ALP, LDH and CK), lipoproteins and hemoglobin. Serum protein electrophoresis (SPE) is a very commonly used analytical method in clinical chemistry. Changes in the relative concentration of fractions allow easy recognition of pathological disorders associated with nephrotic syndrome, inflammatory reaction and hepatic diseases. SPE is a screening test for detecting the M component (MC). Immunofixation (IFE) with use of specific antisera allows detection of the type of MC. SPE is also a method for the quantification of MC and monitoring of disease that is essential for clinical evaluation and follow-up of patients with plasma cell disorders.
Elektroforeza na agaroznom gelu je pouzdana tehnika koja se rutinski koristi u kliničkim laboratorijama za skrining abnormalnosti proteina u različitim biološkim tečnostima (serum, urin, cerebrospinalna tečnost). Zasniva se na principima zonske elektroforeze. Na elektroforetogramima se vizuelno može utvrditi prisustvo kvalitativno ili kvantitativno abnormalnih proteinskih nizova. Tehnika se koristi za elektroforezu seruma, urina, proteina u cerebrospinalnoj tečnosti, enzima (ALP, LDH i CK), lipoproteina i hemoglobina. Elektroforeza proteina u serumu (SPE) vrlo često se kao analitička metoda primenjuje u kliničkoj hemiji. Promene u relativnoj koncentraciji frakcija omogućavaju lako prepoznavanje patoloških poremećaja povezanih s nefrotskim sindromom, inflamatornom reakcijom i oboljenjima jetre. SPE predstavlja skrining test za otkrivanje M komponente (MC). Imunofiksacija uz upotrebu specifičnih antiseruma omogućava detekciju tipa MC. SPE je i metod za kvantifikaciju MC i praćenje toka bolesti što je neophodno za kliničku evaluaciju i praćenje pacijenata sa plazma-ćelijskim bolestima.
Background: In October 2010 an outbreak of staphylococcal food poisoning occurred in an elementary school with a kindergarten in Gorenjska. Methods: Using a questionnaire we performed a retrospective ...cohortanalytical epidemiological study. We calculated attack rate (AR) and relative risk for each food item. In patients we cultivated stools and vomit. We performed an onsite audit where the food samples, environmental samples, and swabs of the hands of the cooks were taken. We performed medical examination of those employed in the kitchen where specimens were taken for microbiological examination. Staphylococcal enterotoxins were detected with agglutination test SET RPLA (Oxoid) and genotypes determined by the pulsed-field gel electrophoresis (PFGE). Results: Out of 374 exposed 73 subjects got ill. Attack rate in the outbreak was 19,5 %. The most frequent symptoms were vomiting (87,7 %), stomach ache (75,3 %), diarrhea (64,2 %) and nausea ( 59,3 %). The highest relative risk (RR) were found for the meatloaf (RR= 24,2 (95 % CI 12,1-48,5; p<0,001)) and potato salad (RR= 19,4 (95 % CI 10,7 - 35,2; p<0,001)). Enterotoxin A producing S. aureus was isolated from hand of a cook, from potato salad and meatlof, from vomit and stools of patients. These strains had the same antibiotic sensitivity and were genetically closely related (96,3 %). Conclusions: In the article an optimal team approaches of a community epidemiologist and microbiologist in the occasion of outbreak are described. We confirmed epidemiologically related isolates and determined the importance of individual methods.
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