Latentna tuberkulozna infekcija

Latent tuberculosis infection (LTBI) is a subclinical infection with Mycobacterium (M.) tuberculosis. Children and immunosuppressed individuals due to pathologic conditions or therapeutic procedures are at a high risk of reactivation of LTBI to active disease compared to general population and require timely identification. The diagnosis of LTBI is based on the measurement an adaptive immune response against M. tuberculosis. The use of tuberculin skin test (TST) has been associated with a number of interfering factors that cause false-positive or false-negative test results. The new ex vivo whole blood tests determining interferon-gamma (IFN-γ) released from T-lymphocytes (interferon-gamma release assays, IGRAs) upon stimulation with M. tuberculosis specific antigens. The introduction of IGRAs to routine clinical practice has improved the diagnosis of LTBI, but recommendations and guidelines for the diagnosis of LTBI are not consistent between different European countries. Therefore, European centre for disease prevention and control developed guidance document based on the up-to-date scientific evidence. Novel concepts of IGRAs and new studies should be designed so as to provide answers to all open questions.Latentna tuberkulozna infekcija (LTBI) definira se kao supklinička infekcija bakterijom Mycobacterium (M.) tuberculosis. Kod djece i osoba oslabljenog imunološkog sustava, zbog pojedinih patoloških stanja ili terapijskih postupaka, postoji veći rizik za aktivaciju LTBI-ja u aktivnu tuberkulozu u odnosu na opću populaciju, te je takvu infekciju potrebno pravovremeno otkriti. Dijagnostika LTBI-ja temelji se na određivanju imunološke reakcije usmjerene protiv M. tuberculosis. Primjena tuberkulinskog testa otežana je brojnim čimbenicima koji mogu uzrokovati lažno pozitivne i lažno negativne rezultate testa. Novi dijagnostički pristup temelji se na ex vivo testovima iz pune krvi kojima se određuje IFN-γ oslobođen iz T-limfocita (interferon-gamma release assays, IGRAs) nakon podražaja specifičnim antigenima za M. tuberculosis. Primjena IGRAs-a u rutinskoj kliničkoj praksi unaprijedila je dijagnostiku LTBI-ja, ali preporuke i smjernice za dijagnostiku LTBI-ja nisu podudarne u pojedinim europskim zemljama. Zbog toga je Europski centar za prevenciju bolesti i kontrolu pripremio smjernice utemeljene na najnovijim znanstvenim podatcima. Unaprjeđenja IGRAs-a i nova istraživanja treba kreirati tako da se mogu dati odgovori na sva otvorena pitanja

Verification of the Canadian Laboratory Initiative on Paediatric Reference Intervals (CALIPER) reference values in Croatian children and adolescents

Introduction: The aim of this study was to examine whether the Canadian Laboratory Initiative on Paediatric Reference Intervals (CALIPER) could be applied to Croatian children and adolescents. Materials and methods: A total of 295 outpatient healthy children and adolescents of age 1 to 18 were selected using the direct a posteriori sampling method. According to current guidelines, 20 samples were tested for each of a total of 51 reference intervals for ferritin, cortisol, dehydroepiandrosterone sulfate, follicle stimulating hormone, lutein stimulating hormone, prolactin, progesterone, sex hormone binding globulin, thyroid stimulating hormone, total testosterone, total thyroxine and total triiodothyronine. Serum samples were analysed on the Beckman Coulter DxI600 immunoassay analyser by chemiluminescence immunoassay method. A reference interval was adopted if < 10% of the results fall outside CALIPER reference interval range. For analytes in which this criterion is not met in the first set of samples, a new set of 20 samples were collected. Results: After the first set of measurements, 96% of all tested reference intervals were adopted for use. The additional sets of 20 reference subjects were tested for only two reference intervals; follicle stimulating hormone for female aged 1 to 9 years, and irrespective of the gender, sex hormone binding globulin for children aged 8 to 11 years. All results of additional samples were within the specified interval limits. Conclusions: CALIPER reference intervals for ferritin and 11 hormones defined for Beckman Coulter DxI600 immunoassay analyser can be implemented into the Croatian laboratories and clinical practice

Troponin i B-tip natrijuretskog peptida: laboratorijska dijagnostika i klinička upotreba u pedijatriji

For years now, troponin and B-type natriuretic peptide are two cardiac biomarkers routinely used in the diagnosis of cardiac disease in general population, but the knowledge of their usability in pediatric population is very low. This review covers the issues of determination, interpretation of laboratory test results, and usefulness of troponin and B-type natriuretic peptide in pediatrics, based on published data and our own experience. It is intended for pediatricians and laboratory specialists who work with pediatric population.Troponin i B-tip natrijuretskog peptida su dva srčana biljega koja se u općoj odrasloj populaciji već dugi niz godina rutinski primjenjuju u dijagnostici srčanih bolesti, no malo je spoznaja o njihovoj korisnoj upotrebi u pedijatrijskoj praksi. Ovaj pregledni rad obuhvaća svu problematiku određivanja, tumačenja nalaza i korisnost troponina i B-tipa natrijuretskog peptida u pedijatriji na temelju podataka iz literature i vlastitog iskustva. Namijenjen je liječnicima pedijatrima i laboratorijskim stručnjacima koji se u svom radu susreću s dječjom populacijom

Latent tuberculosis infection in a subject with diabetes mellitus - a case report

Uvod: Bolesnici sa šećernom bolesti tipa 1 često imaju narušenu staničnu imunost i podložni su zaraznim bolestima. Cilj je ovog rada prikazati slučaj latentne tuberkulozne infekcije (LTBI) u 46-godišnje zdravstvene djelatnice (medicinske sestre na odjelu za tuberkulozu male djece) koja 20 godina boluje od šećerne bolesti tipa 1. Materijali i metode: Tuberkulinski kožni test učinjen je s 2 tuberkulinske jedinice (Tuberkulin RT-23, Statens Serum Institute, Kopenhagen, Danska). Krv za ispitivanje oslobađanja interferona gama, IFN-γ, (engl. Interferon Gamma Releasing Assay, IGRA) uzorkovana je u epruvete koje sadrže specifične antigene mikobakterija tuberkuloze (ESAT-6, CFP10 i TB7.7). Nakon inkubacije (22 sata) na 37°C u plazmi je određena koncentracija IFN-γ metodom ELISA (Qu-antiFERON-TB Gold In Tube - Cellestis Ltd., Victoria, Australia). Pozitivna kontrola bila je krv uzorkovana s mitogenom fitohemaglutininom, a negativna kontrola krv uzorkovana s heparinom. Rezultati: Tuberkulinski kožni test bio je negativan (induracija 6 mm), a nalaz otpuštanja IFN-γ iz aktiviranih limfocita (≥ 0,35 kIU/L) pozitivan. Normalna stanična imunost potvrđena je pozitivnom kontrolom. Ispitanica nema znakova aktivne tuberkuloze. Zaključak: U medicinske sestre koja boluje od šećerne bolesti tipa 1, a godinama radi s tuberkuloznim bolesnicima LTBI je dokazana testom oslobađanja IFN-γ, ali ne i tuberkulinskim kožnim testom. Unatoč dugogodišnje šećerne bolesti tipa 1, reakcija oslobađanja IFN-γ prema specifičnim tuberkuloznim antigenima je očuvana, a aktivna se tuberkuloza nije razvila.Introduction: Patients suffering from diabetes mellitus type 1 often have impaired cell-mediated immunity and are prone to infectious diseases. The aim of this paper is to present a case study on latent tuberculosis infection (LTBI) in a 46 year-old female health care worker (a nurse at the department for tuberculosis in young children) who has been having type 1 diabetes for 20 years. Materials and methods: Tuberculin skin test was performed using 2 tuberculin units (Tuberkulin RT-23, Statens Serum Institute, Copenhagen, Denmark). Blood for interferon gamma (IFN-γ) release assay, (IGRA) was sampled in test tubes containing specific antigens of mycobacterium tuberculosis (ESAT-6, CFP10 and TB7.7). After incubation (22 hours) at 37°C, IFN-γ concentration in plasma was determined by ELISA assay (QuantiFERON-TB Gold In Tube - Cellestis Ltd., Victoria, Australia). For positive control, blood was sampled by mitogene phytohemagglutinin and for negative control by heparin. Results: Tuberculin skin test was negative (induration 6 mm) and test result of IFN- γ release from activated lymphocytes (≥ 0.35 kIU/L) positive. Normal cellular immunity was confirmed by positive control sample. The subject showed no signs of active tuberculosis. Conclusion: In a nurse suffering from type 1 diabetes who has worked for years with tuberculosis patients, LTBI was detected by IFN-γ release assay rather than by tuberculin skin test. In spite of the long lasting diabetes, reaction of IFN-γ release toward specific tuberculosis antigens has been preserved and active tuberculosis has not developed

Hepatitis during respiratory syncytial virus infection – a case report

Introduction: Respiratory syncytial virus (RSV) infection is the most common cause of hospitalization in infants and small children. The aim was to present a 13-months old boy diagnosed with acute airway infection, acute otitis media (AOM) and hepatitis during the RSV-infection. Material and methods: Serum catalytic activities of alkaline phosphatase (ALP), aspartate aminotranspherase (AST), alanine aminotranspherase (ALT), gamma glutamyl transpherase (GGT), lactate dehydrogenase (LD), and concentrations of bilirubin were monitored during hospitalization and at control examination. Results: The child had clinical signs and symptoms of respiratory failure, AOM, and laboratory findings of virus infection and liver disease. On admission, catalytic activities of enzymes were markedly increased, especially the activity of ALP (10333 U/L, i.e. 24-fold increase in comparison with the upper reference limit). The highest increased in AST (339 U/L, 4.5-fold), ALT (475 U/L, 10.3-fold) and LD (545 U/L, 1.5-fold) were registered on the 3rd day, and the highest increase in GGT (68 U/L, 3.1-fold) occurred on the 11th day. Seven weeks after discharge AST, ALT, GGT and LD decreased into reference range, and ALP remain mildly increased (478 U/L, 1.1 fold increase). RSV was confirmed in nasal lavage fluid. Conclusion: Laboratory results in patient with RSV infection needs to be interpreted in the light of both, respiratory and extrapulmonary manifestations of the infection, respectively

Increased IgA in a subject with positive anti-thyroid autoantibodies – a case report

Uvod: Povećane koncentracije antitireoidnih autoantitijela nalaze se u upalnim bolestima te u autoimunim bolestima štitnjače. Cilj ovog članka jest prikazati povećane vrijednosti IgA u ispitanice s pozitivnim antitireoidnim autoantitijelima. Navode se rezultati trinaestomjesečnog praćenja. Materijali i metode: Koncentracije tireoidnih hormona i antitijela u uzorcima seruma izmjerene su primjenom enzimske imunoanalize na mikročesticama. IgA je određen pomoću dvije imunokemijske metode, tj. imunoturbidimetrijskom metodom i radijalnom imunodifuzijom. Rezultati: Početne pretrage funkcije štitnjače dale su normalne vrijednosti T3, T4 i hormona koji stimulira štitnjaču (TSH) te povećane vrijednosti autoantitijela na tireoidnu peroksidazu (anti-TPO), tireoglobulin (anti-Tg) i IgA. Ispitanica je liječena jednokratnim dnevnim dozama levotiroksina. Povećane početne koncentracije anti-TPO stabilizirane su primjenom levotiroksina, dok su anti-Tg ostala povećana. IgA je mjeren imunoturbidimetrijskom metodom i bio je značajno povećan tijekom razdoblja praćenja ispitanice. Ponovljenim određivanjem IgA radijalnom imunodifuzijom otkrivene su manje vrijednosti IgA. Zaključak: Moguće lažno povećane vrijednosti IgA mogu biti rezultat, kako interferencija kozjih anti-IgA antitijela iz reagensa, tako i križne reaktivnosti s endogenim anti-Tg autoantitijelima u serumu. Potrebna su daljnja istraživanja kako bi se utvrdio uzrok povećanih vrijednosti IgA u bolesnice s pozitivnim antitireoidnim autoantitijelima.Introduction: Increased concentrations of antithyroid autoantibodies are found in inflammatory diseases as well as in thyroid autoimmune disorders. The aim of this report is to present increased IgA concentrations in a female patient with positive anti-thyroid autoantibodies. Results throughout 13 months of follow-up are presented. Materials and methods: The levels of thyroid hormones and antibodies in the serum samples were measured using microparticle enzyme-immunoas-say. IgA was determined by two immunochemical methods, immuno-turbi-dimetric method and radial immunodiffusion, respectively. Results: Initial thyroid function tests showed normal values of T3, T4 and thyroid stimulating hormone (TSH), and increased values of thyroid peroxi-dase autoantibodies (anti-TPO), anti-thyroglobulin (anti-Tg) autoantibodies and IgA. The patient underwent treatment with single daily dose of levot-hyroxine. The elevated initial levels of anti-TPO stabilized with the administration of levothyroxine, while anti-Tg remained increased. IgA determined by immuno-turbidimetric method was significantly increased during the follow-up period. Repeated determination of IgA by radial immunodiffusion revealed lower IgA concentrations. Conclusion: Possible false increased IgA values could be a result of both, interferences of goat anti-IgA antibodies from reagents, and cross-reactivity with endogenous anti-Tg autoantibodies in serum, respectively. Further investigations are needed to ascertain the cause of increased IgA values in patient with positive anti-thyroid autoantibodies

Correlation between serum butyrylcholinesterase activity and serum lipid concentrations in rats treated with different antagonists of the adrenergic system

Background and Purpose: Based on the facts that the blockade of adrenergic receptors can alter lipid profile in the serum and that it has been suggested that butyrylcholinesterase (BuChE) is involved in lipid metabolism, different adrenergic blocking agents were administered to rats to modify lipid concentrations in serum. The activity of BuChE was examined under such conditions and correlations with serum lipids were investigated. The purpose of this study was to evaluate the effects of different adrenergic antagonists on BuChE activity and to investigate the correlation between BuChE activity and serum lipids. Materials and Methods: Six groups of male Fischer 344 rats (9 animals/ group) were treated orally with adrenergic antagonists (mixed in commercial diet) during 6 weeks: oxprenolol, atenolol, doxazosin, oxprenolol and doxazosin, atenolol and doxazosin, and guanethidine. A control group (9 rats) received only commercial diet. BuChE activity in serum was determined with kinetic color test using butyrylthiocholine as a substrate. Concentrations of serum lipids (total cholesterol, triglycerides and HDL cholesterol) were determined by enzymatic colorimetric tests. Data were analyzed by Kruskal-Wallis test and Spearman’s correlation coefficient. Results: The results revealed that oxprenolol and doxazosin (given alone or in combination with atenolol or oxprenolol) increased (>30 %) BuChE activity. BuChE activity correlated with different serum lipids, and correlation depended on the type of adrenergic blockade. Conclusion: Although the examined adrenergic antagonists did not influence serum lipid concentrations, the increase of BuChE activity and correlation with serum lipid concentrations suggested that the increase of this enzyme’s activity might be the first sign of altered lipid metabolism

Fecal Calprotectin as a Biomarker of Food Allergy and Disease Severity in Children with Atopic Dermatitis without Gastrointestinal Symptoms

Fecal calprotectin (FCP) is a biomarker of intestinal inflammation and has recently been proposed as a diagnostic biomarker of food allergy (FA) in children. The aim of this study was to compare FCP level in infants and children under 4 years old with 1) atopic dermatitis (AD) with food allergy (FA) and 2) children with AD and without FA with the results in healthy controls. In total, 46 infants and children (mean age 14 months ± 12) diagnosed with AD were divided into two groups: G1, children with atopic AD with FA (n=28) and G2, children with AD without FA (n=18). The control group (G3) was made up of healthy children of the same age (n=18). The median FCP was significantly higher in G1 compared with G2 (G1: median 154, IQR 416 μg/g vs G2: median 41.3, IQR 59 μg/g; P=0.0096). The median FCP in children with AD and FA was significantly higher before elimination diet in comparison with FCP after 3 months of elimination diet (median 154, IQR 416 μg/g vs median 35, IQR 23 μg/g; P=0.0039). The level of FCP was significantly positively correlated with the SCORAD score (r=0.5544, P=0.0022). Our study showed a significant difference in level of FCP in patients with AD without FA compared with patients with AD and FA. We also found a positive correlation of FCP with SCORAD score, a biomarker of AD severity. New studies are needed to investigate the role of FCP as a biomarker of FA in children with AD

Fecal Calprotectin as a Biomarker of Food Allergy and Disease Severity in Children with Atopic Dermatitis without Gastrointestinal Symptoms

Fecal calprotectin (FCP) is a biomarker of intestinal inflammation and has recently been proposed as a diagnostic biomarker of food allergy (FA) in children. The aim of this study was to compare FCP level in infants and children under 4 years old with 1) atopic dermatitis (AD) with food allergy (FA) and 2) children with AD and without FA with the results in healthy controls. In total, 46 infants and children (mean age 14 months ± 12) diagnosed with AD were divided into two groups: G1, children with atopic AD with FA (n=28) and G2, children with AD without FA (n=18). The control group (G3) was made up of healthy children of the same age (n=18). The median FCP was significantly higher in G1 compared with G2 (G1: median 154, IQR 416 μg/g vs G2: median 41.3, IQR 59 μg/g; P=0.0096). The median FCP in children with AD and FA was significantly higher before elimination diet in comparison with FCP after 3 months of elimination diet (median 154, IQR 416 μg/g vs median 35, IQR 23 μg/g; P=0.0039). The level of FCP was significantly positively correlated with the SCORAD score (r=0.5544, P=0.0022). Our study showed a significant difference in level of FCP in patients with AD without FA compared with patients with AD and FA. We also found a positive correlation of FCP with SCORAD score, a biomarker of AD severity. New studies are needed to investigate the role of FCP as a biomarker of FA in children with AD

A new case of transient hyperphosphatasemia in a 21 -month-old child with recurrent wheezing - case repor

Uvod: Opisuje se slučaj prolazne hiperfosfatazemije u 21-mjesečnog dječaka s ponavljajućom sipnjom tijekom akutnog respiracijskog infekta. Materijali i metode: Određivanje katalitičke aktivnosti alkalne fosfataze provedeno je preporučenom fotometrijskom metodom, a elektroforetsko razdvajanje izoenzima alkalne fosfataze na agaroznom gelu uz obradu seruma lektinom. Rezultati: Tijekom bolničkog liječenja dijete je imalo povećanu katalitičku aktivnost alkalne fosfataze: prvog dana 4109 U/L, šestog dana 2156 U/L. Pri kontrolnom pregledu 27. dana od prijma u bolnicu aktivnost se smanjila na 505 U/L. Elektroforetskim razdvajanjem izoenzima alkalne fosfataze dokazana je elektroforetska vrpca brža od jetreno-koštanog izoenzima, tzv. brza anodna vrpca. Zaključak: Pojava izrazito povećane katalitičke aktivnosti alkalne fosfataze uz tipičnu elektroforetsku razdiobu izoenzima u trajanju od nekoliko tjedana ukazuje na prolaznu hiperfosfatazemiju. Pravodobnim prepoznavanjem izbjegava se daljnja nepotrebna dijagnostička obrada.Introduction: A case of transient hyperphosphatasemia in a 21-month-old boy diagnosed with recurrent wheezing during acute respiratory infection is described. Materials and methods: Determination of alkaline phosphatase catalytic activity was performed by the recommended photometric method, and elec-trophoretic separation of alkaline phosphatase isoenzymes on agarose gel by serum processing with lectin. Results: During hospital treatment, increased catalytic activity of alkaline phosphatase was recorded (4109 U/L on day 1 and 2156 U/L on day 6). On control examination on day 27 of hospital admission, the value decreased to 505 U/L. Electrophoretic separation of alkaline phosphatase isoenzymes demonstrated a band faster than the hepatic-osseous isoenzyme, a fast anodal form. Conclusion: A marked increase in the alkaline phosphatase catalytic activity with typical electrophoretic isoenzyme pattern persisting for several weeks points to transient hyperphosphatasemia. Additional unnecessary diagnostic work-up can be obviated by timely recognition of the phenomenon