TREATMENT OF SEPSIS IN NEUTROPENIC PATIENTS

Febrilna neutropenija je česta i potencijalno fatalna komplikacija intenzivne citostatske terapije zbog čega je glavni uzrok nerelapsnog mortaliteta hematoloških pacijenata. Važnu skupinu pacijenata čine oni koji su zbog osnovne bolesti ili ranijih linija liječenja pod rizikom za infekcije specifi čnim uzročnicima. Kako se u većine pacijenata s febrilnom neutropenijom može postaviti dijagnoza sepse, dijagnostičke i terapijske postupke treba primjenjivati kao da je pacijent u sepsi. Bitno je prepoznati visokorizične pacijente jer se incidencija sepse u neutropeničnih pacijenata može znatno smanjiti adekvatnom antimikrobnom profilaksom i granulocitnim faktorima rasta. Ako dođe do sepse, vrlo je važno što ranije započeti liječenje prema ustanovljenim smjernicama, ali uvijek uzimajući u obzir i individualne karakteristike svakog pacijenta kao i mikrobiološku situaciju, čime se može znatno smanjiti mortalitet i morbiditet ovih pacijenata.Febrile neutropenia is a common and potentially fatal complication of intense cytotoxic therapy, which makes it the main cause of non-relapse mortality in patients with hematologic malignancies. Some of the patients are at risk of specifi c infections due to underlying disease of previous treatment regimens. Considering that most febrile neutropenic patients can be diagnosed with sepsis, diagnosis and treatment should be treated as septic. It is important to recognize patients at high risk since the incidence of sepsis in neutropenic patients can be signifi cantly reduced using antimicrobial prophylaxis and granulocyte growth factors. Once sepsis occurs, prompt treatment according to guidelines, individualized based on the specifi c problems of each patient, and microbiological situation can significantly reduce mortality and morbidity

In vitro uzgoj limbalnog presatka svinje

The stem cells that provide tissue regeneration are residents of different organ structures in the body and are usually located in well-protected sites of organs and tissues. For stem cells of either corneal epithelium such sites are in the basal epithelial layer of the cornea residing in its limbal region, termed the palisade of Voight. The growing interest in allografts and xenografts implies a thorough study of the regenerative potential of these cells, as well as a clear description of their patterns in the in vitro tissue cultures to be grafted. The aim of this study was to evaluate the culturing patterns of porcine limbal stem cells and to optimize growing conditions of these cell cultures, in order to develop a reliable biomedical model intended for studying the potentials of allografts/xenografts originated from porcine tissues. Porcine stem cell equivalents have potential in reparative/regenerative veterinary medicine for pets, horses and selected breeds of food animals, but also as xenografts for humans.Zametne stanice, koje omogućuju regeneraciju tkiva, nalaze se u različitim organskim sustavima u tijelu i obično su smještene u dobro zaštićenim dijelovima organa i tkiva. Za zametne stanice rožnice takva su mjesta u bazalnom sloju epitela rožnice, njezinoga limbalnoga područja nazvanoga Voightove palisade. Rastuće zanimanje za alotransplantate i ksenotransplantate predmnijeva temeljita istraživanja regeneracijskog potencijala tih stanica, kao i defi niranje njihovih osobitosti u in vitro kulturama za presađivanje. Cilj ovoga istraživanja bio je vrednovanje osobitosti uzgajanja svinjskih limbalnih zametnih stanica u kulturi i optimiziranje uvjeta njihovoga uzgoja radi uspostavljanja pouzdanoga biomedicinskoga modela pogodnoga za istraživanje potencijala alotransplantata i ksenotransplantata podrijetlom od svinjskih tkiva. Svinjske zametne stanice imaju potencijal u reparativnoj/regenerativnoj veterinarskoj medicini za kućne ljubimce, konje i visokoselekcionirane pasmine životinja namijenjenih ljudskoj prehrani, ali također i u ksenotransplantaciji za čovjeka

Vrijednosti hematoloških pokazatelja u pasa liječenih supkonjunktivalnom primjenom staničnog alografta.

The cornea, as a source for obtaining stem cells, is a good medium for their use in veterinary ophthalmology. In this research, we used dogs which had been diagnosed with extensive, chronic thinning of the central cornea (vertex corneae) generated due to complicated mechanical damage to the corneae pannosa. The goal of this study was to test whether subconjuctival application of limbal stem cells allografts had any influence on haematology parameters in peripheral blood. The animals were divided into three experimental groups regardless of breed, sex and age, and each group comprised 7 dogs. Group A consisted of clinically healthy dogs (control), in group B there were dogs who had had injuries treated by a conventional ophthalmological approach in veterinary medicine, and group C in which corneal injuries were treated using subconjuctival cell allografts. Blood samples were taken four times during the experiment from all animals for haematological analysis. Transplantation of limbal stem cell allografts did not affect the physiological haematology parameters.Rožnica, kao izvor matičnih stanica, dobar je medij za njihovu uporabu u veterinarskoj oftalmologiji. U ovom istraživanju koristili smo pse kojima su dijagnosticirana opsežna, kronična stanjivanja središnjeg dijela rožnice, posljedica mehaničkih oštećenja, dodatno komplicirana upalnim procesima. Cilj istraživanja bio je ispitati utjecaj supkonjunktivalne primjene limbalnog staničnog alografta na hematološke pokazatelje u perifernoj krvi pacijenata. Životinje su bile podijeljene u tri pokusne skupine, bez obzira na pasminu, spol i dob. U svakoj skupini bilo je sedam pasa. U skupini A bili su klinički zdravi psi (kontrolna skupina), u skupini B bili su psi kojima su ozljede obrađene konvencionalnim oftalmološkim pristupom, a psima u skupini C oštećenja rožnice bila su liječena supkonjunktivalnom primjenom staničnog alografta. Četiri puta tijekom istraživanja svim su životinjama uzeti uzorci periferne venske krvi za hematološke pretrage. Supkonjunktivalna primjena staničnog alografta nije utjecala na promjenu vrijednosti hematoloških pokazatelja u obrađivanih pasa

Supkonjunktivalna primjena alogeničnih limbalnih stanica u pasa s poremećajima rožnice.

Limbal stem cells play a crucial role in maintaining the integrity of the corneal surface in a healthy condition or after injury through corneal renewal and repair. In pathologic conditions these cells could be replenished with cultivated allogenic limbal cells. In this investigation, limbal stem cells from small fragments of donor tissue were cultivated in vitro for treatment of corneal lesions in dogs. Fourteen dogs were divided into two groups of seven animals each: a group of dogs with corneal lesions treated using the classical veterinary ophthalmology approach, and a group of dogs with corneal lesions treated with the application of cultivated allogenic limbal cells. Regardless of the size and location of the corneal lesions, after 30 days in the group of dogs with corneal lesions treated with the application of cultivated allogenic limbal cells ophthalmological examination showed no signs of any eye disorder except corneal edema (14.29 %). In the group of dogs with corneal lesions cured using the classical veterinary ophthalmology approach, 30 days after the beginning of treatment ophthalmological examination showed 42.86 % mild/incisional blepharospasm, 85.71 % secretion, 100 % observable conjuctival congestion, 14.29 % eye liquor dimming, 85.71 % corneal edema and 85.71 % irritation. After application of cultivated allogenic limbal cells, corneal lesions were completely healed 15 days earlier than the lesions treated with conventional ophthalmological therapy. Allogenic limbal cells were capable of restoring corneal clarity with no development of ocular complications.Limbalne matične stanice igraju ključnu ulogu u održavanju integriteta površine rožnice u zdravom stanju ili nakon ozljede, kroz obnavljanje i popravak rožnice. U patološkim stanjima te se stanice mogu nadomijestiti s uzgojenim alogeničnim limbalnim stanicama. U ovom istraživanju limbalne matične stanice iz malih fragmenata tkiva donora uzgojene su in vitro radi liječenja ozljeda rožnice u pasa. Četrnaest pasa bilo je podijeljeno u dvije skupine od sedam životinja: skupina pasa s ozljedama rožnice liječenim klasičnim veterinarskim oftalmološkim pristupom i skupina pasa s ozljedama rožnice liječenim primjenom in vitro uzgojnih alogeničnih limbalnih stanica. Neovisno o veličini i smještaju lezije rožnice, deset dana nakon primjene stanica, oftalmološki pregled je pokazao 80% manje izražene simptome blefarospazma, 85% smanjenu sekreciju, 43% manje uočljivu konjunktivalnu kongestiju, 31% manji edem rožnice i 55% smanjenu iritaciju u usporedbi s psima liječenim klasičnim oftalmološkim pristupom, bez znakova zamućenja očne vodice ili priraslica. Nakon primjene uzgojeni alogeničnih limbalnih stanica, lezije rožnice u potpunosti su bile izliječene 15 dana ranije nego lezije liječene konvencionalnim oftalmološkim načinom. Supkonjunktivalna primejna limbalnih stanica omogućuje potpunu regeneraciju ozljeda rožnice nakon 30 dana bez obzira na veličinu i opseg defekta. Alogenične limbalne stanice sposobne su obnoviti prozirnost rožnice bez razvoja komplikacija. Iako su postignuti rezultati bili ohrabrujući, potrebna su daljnja istraživanja za optimizaciju protokola liječenja

Vrijednosti hematoloških pokazatelja u pasa liječenih supkonjunktivalnom primjenom staničnog alografta.

The cornea, as a source for obtaining stem cells, is a good medium for their use in veterinary ophthalmology. In this research, we used dogs which had been diagnosed with extensive, chronic thinning of the central cornea (vertex corneae) generated due to complicated mechanical damage to the corneae pannosa. The goal of this study was to test whether subconjuctival application of limbal stem cells allografts had any influence on haematology parameters in peripheral blood. The animals were divided into three experimental groups regardless of breed, sex and age, and each group comprised 7 dogs. Group A consisted of clinically healthy dogs (control), in group B there were dogs who had had injuries treated by a conventional ophthalmological approach in veterinary medicine, and group C in which corneal injuries were treated using subconjuctival cell allografts. Blood samples were taken four times during the experiment from all animals for haematological analysis. Transplantation of limbal stem cell allografts did not affect the physiological haematology parameters.Rožnica, kao izvor matičnih stanica, dobar je medij za njihovu uporabu u veterinarskoj oftalmologiji. U ovom istraživanju koristili smo pse kojima su dijagnosticirana opsežna, kronična stanjivanja središnjeg dijela rožnice, posljedica mehaničkih oštećenja, dodatno komplicirana upalnim procesima. Cilj istraživanja bio je ispitati utjecaj supkonjunktivalne primjene limbalnog staničnog alografta na hematološke pokazatelje u perifernoj krvi pacijenata. Životinje su bile podijeljene u tri pokusne skupine, bez obzira na pasminu, spol i dob. U svakoj skupini bilo je sedam pasa. U skupini A bili su klinički zdravi psi (kontrolna skupina), u skupini B bili su psi kojima su ozljede obrađene konvencionalnim oftalmološkim pristupom, a psima u skupini C oštećenja rožnice bila su liječena supkonjunktivalnom primjenom staničnog alografta. Četiri puta tijekom istraživanja svim su životinjama uzeti uzorci periferne venske krvi za hematološke pretrage. Supkonjunktivalna primjena staničnog alografta nije utjecala na promjenu vrijednosti hematoloških pokazatelja u obrađivanih pasa

Prognostic factors in myelodysplastic syndromes: single-center experience

Uvod: Mijelodisplastični sindrom (MDS) jedna je od najčešćih klonskih hematoloških neoplastičnih neoplazma starije dobi karakterizirana jednom ili s više citopenija te ograničenim terapijskim rješenjima. Cilj je ovog rada bila retrospektivna analiza klasičnih prognostičkih čimbenika kod bolesnika dijagnosticiranih u jednom centru. Kao glavni ishod uzeto je ukupno preživljenje (engl. Overall survival – OS) definirano kao smrt od posljedica same bolesti ili nekoga drugog uzroka. Metode: Retrospektivno smo analizirali povijesti bolesti svih bolesnika dijagnosticiranih u jednom centru u razdoblju od 1. siječnja 2013. do 31. prosinca 2016. godine. Rezultati: U istraživanje je uključeno ukupno 58 bolesnika s medijanom dobi od 69 godina. Nakon medijana praćenja od 12 mjeseci medijan OS-a iznosio je 17 mjeseci s procijenjenom trogodišnjom stopom OS-a od 25%. U analizi prognostičkih čimbenika zbrojevi rizične stratifikacije prema IPSS-u, R-IPSS-u i WPSS-u bili su statistički značajni te su jasno diskriminirali OS ispitanika s nižim rizikom u odnosu prema OS-u ispitanika s visokom rizikom, no zbog malenog broja bolesnika nije bilo moguće odgovoriti na pitanje koji je bodovni sustav najprikladniji u ovoj populaciji. Citogenetičke kategorije prema IPSS-u i R-IPSS-u bile su statistički značajni prediktori OS-a. Usprkos statističkoj tendenciji morfološke karakteristike (podtip MDS-a, broj blasta u koštanoj srži) nisu se pokazale statistički značajnim čimbenicima prognoze u ovih bolesnika. Od kliničkih karakteristika jedino su prisutnost anemije i njezina težina, odnosno transfuzijska ovisnost bile znatno povezane s lošijim ishodom. Zaključak: Na našoj kohorti bolesnika većina tradicionalnih faktora pokazala se važnom u skladu s literaturnim podacima.Introduction: Myelodysplastic syndrome (MDS) is one of the most common myeloid neoplasms of elderly characterized by cytopenias and limited therapeutic options. The main aim of this retrospective singlecenter study was to examine the value of classical prognostic factors. The main outcome of the study was overall survival (OS) defined as death from MDS or any other reason. Methods: We analyzed the medical records of patients diagnosed with MDS at single centre in the period from beginning of 2013 to the end of 2016. Results: Total of 58 patients (median age of diagnosis being 69 years) were included in the study. After median of follow-up of 12 months, median OS was 17 months and estimated 3-year OS rate 25%. Classical prognostic systems such as IPSS, WPSS and R-IPSS were statistically significant prognostic factors discriminating adequately between low and high risk groups in terms of outcome. However , due to small sample size, we were not able to distinguish the most appropriate scoring system. The cytogenetics subgroups according to IPPS and R-IPSS were significant predictors of outcomes underlying its crucial role in MDS diagnosis. Despite the statistical tendency morphological features of MDS (2008 World Health Organization subtype and number of blasts in bone marrow) were not significant predictors of OS. Among clinical features, only presence and degree of anemia and transfusion dependency were significant predictors of inferior survival. Conclusion: The majority of traditional prognostic factors were significant in our cohort in concordance with literature review

AUTOLOGOUS STEM CELL TRANSPLANTATION IN REFRACTORY OR RELAPSED DIFFUSE LARGE B CELL LYMPHOMA – A SINGLE CENTRE EXPERIENCE

Autologna transplantacija perifernih matičnih stanica zlatni je standard u liječenju refraktornog ili relapsnoga kemosenzitivnog difuznog B-velikostaničnog limfoma u pogodnih bolesnika. Cilj je ovog rada prikazati ishode autologne transplantacije perifernih matičnih stanica u bolesnika s refraktornim ili relapsnim ne-Hodgkinovim limfomom difuznoga B-velikostaničnog tipa. Retrospektivno smo analizirali podatke 62-je bolesnika liječenih autolognom transplantacijom u našem centru u razdoblju od 2000. do 2013. godine. Većina bolesnika (71%) liječena je kemoterapijom miniBEAM, a svi su primili mijeloablativnu kemoterapiju BEAM s reinfuzijom vlastitih matičnih stanica. Ukupna stopa odgovora (kom­pletna i parcijalna remisija) nakon autologne transplantacije iznosila je 75,8%. Medijan ukupnog preživljenja iznosio je 37,2 mjeseca. Medijan preživljenja bez događaja vezanih uz bolest iznosio je 16,9 mjeseci. Čimbenici statistički značajno povezani s ukupnim preživljenjem bili su aktivnost bolesti prije visokodozne spasonosne terapije, odgovor na spasonosnu terapiju. Internacionalni prognostički indeks, stadij bolesti, odgovor na autolognu transplantaciju perifernih matičnih sta­nica te radioterapija nakon autologne transplantacije. Liječenje rituksimabom nije bilo statistički značajno povezano s ishodom. U ovoj skupini bolesnika autologna je transplantacija perifernih matičnih stanica bila učinkovita u postizanju remisije i preživljenja, što je dobar i očekivan ishod navedenog postupka. Naglašavamo da je i u skupini bolesnika s kemorezistentnom bolešću autologna transplantacija bila učinkovita u 32,5% bolesnika.Autologous stem cell transplantation represents the gold standard in chemosensitive diffuse B large cell lymphoma in relapse or in refractory setting. The aim of this study was to present the outcome of peripheral autologous stem cell transplantation in patients with refractory or relapsed diffuse large B cell lymphoma. We retrospectively analysed the data of 62 patients, who underwent this procedure for the period 2000–2013. The majority of patients (71%) were treated with miniBEAM salvage chemotherapy and all received BEAM myeloablative protocol followed by the stem cell reinfusion. The overall response rate for autologous transplantation was 75.8%. Median overall survival was 37.2 months. Median event-free survival was 16.9 months. Factors associated with overall survival were state of disease prior to salvage chemotherapy, chemosensitivity of disease, International prognostic index, disease activity at the relapse, response to autologous transplantation and post-transplantation radiotherapy. The use of rituximab was not significantly correlated to the outcome. In this patient group autologous stem cell transplantation was found to be effective in achieving remission and survival showing the adequate role of this procedure in this clinical setting. We stress out that autologous stem cell transplantation was effective in 32.5% patients with chemorefractory disease after salvage therapy

Smjernice za dijagnozu i liječenje bolesnika s mijelodisplastičnim sindromom [Guidelines for diagnosis and treatment of patients with myelodysplastic syndrome]

Myelodysplastic syndrome Working Group of the Croatian Cooperative Group for Hematologic Diseases (CROHEM), Referral center of the Ministry of Health of the Republic of Croatia for diagnostics and treatment of MDS, as well as the Croatian Society for Haematology of the Croatian Medical Association have made Croatian guidelines for diagnosis and treatment of myelodysplastic syndrome (MDS). MDS is a heterogeneous group of clonal hematopoietic stem cell disorders characterized by ineffective hematopoiesis, dysplasia, cytopenia and risk of transformation to acute myeloid leukemia (AML). Diagnosis is based on morphological characteristics of hematopoietic cells supplemented with the cytogenetic analysis and bone marrow flow cytometry. Due to great differences in the natural course of the disease, i.e. time to progression to AML and the expected time of survival several scoring systems have been developed to determine the disease risk. The treatment of patients with MDS is based on the risk factors of the disease as well as the individual risk of treatment

GUIDELINES FOR DIAGNOSIS AND TREATMENT OF PATIENTS WITH MYELODYSPLASTIC SYNDROME

Radna skupina za mijelodisplastični sindrom Hrvatske kooperativne grupe za hematološke bolesti (KROHEM), Referalni centar Ministarstva zdravlja Republike Hrvatske za dijagnostiku i liječenje mijelodisplastičnog sindroma te Hrvatsko hematološko društvo Hrvatskoga liječničkog zbora izradili su hrvatske smjernice za dijagnozu i liječenje mijelodisplastičnog sindroma (MDS). MDS je heterogena grupa klonskih bolesti matične hematopoetske stanice karakteriziranih neučinkovitom hematopoezom, displazijom, citopenijom i rizikom od transformacije u akutnu mijeloičnu leukemiju (AML). Dijagnoza se postavlja na temelju morfoloških karakteristika stanica hematopoeze uz nadopunu citogenetskom analizom i imunofenotipizacijom koštane srži. Zbog velikih razlika u prirodnom tijeku bolesti, odnosno u vremenu do progresije u AML i u očekivanom preživljenju razvijeno je više sustava bodovanja kojima se određuje stupanj rizika od bolesti. Liječenje pacijenta s MDS-om temelji se na stupnju rizika od bolesti i individualnom riziku od liječenjaMyelodysplastic syndrome Working Group of the Croatian Cooperative Group for Hematologic Diseases (CROHEM), Referral center of the Ministry of Health of the Republic of Croatia for diagnostics and treatment of MDS, as well as the Croatian Society for Haematology of the Croatian Medical Association have made Croatian guidelines for diagnosis and treatment of myelodysplastic syndrome (MDS). MDS is a heterogeneous group of clonal hematopoietic stem cell disorders characterized by ineffective hematopoiesis, dysplasia, cytopenia and risk of transformation to acute myeloid leukemia (AML). Diagnosis is based on morphological characteristics of hematopoietic cells supplemented with the cytogenetic analysis and bone marrow fl ow cytometry. Due to great differences in the natural course of the disease, i.e. time to progression to AML and the expected time of survival several scoring systems have been developed to determine the disease risk. The treatment of patients with MDS is based on the risk factors of the disease as well as the individual risk of treatment

Prognostic impact of a suboptimal number of analyzed metaphases in normal karyotype lower-risk MDS

Conventional karyotype is one of the most relevant prognostic factors in MDS. However, about 50% of patients with MDS have a normal karyotype. Usually, 20-25 normal metaphases (nMP) are considered to be optimal to exclude small abnormal clones which might be associated with poor prognosis. This study evaluated the impact of examining a suboptimal number of metaphases in patients recruited to the EUMDS Registry with low and intermediate-1 risk according to IPSS. Only 179/1049 (17%) of patients with a normal karyotype had a suboptimal number of nMP, defined as less than 20 metaphases analyzed. The outcome (overall survival and progression-free survival) of patients with suboptimal nMP was not inferior to those with higher numbers of analyzed MP both in univariate and multivariate analyses. For patients with an abnormal karyotype, 224/649 (35%) had a suboptimal number of MP assessed, but this did not impact on outcome. For patients with a normal karyotype and suboptimal numbers of analyzable metaphases standard evaluation might be acceptable for general practice, but we recommend additional FISH-analyses or molecular techniques, especially in candidates for intensive interventions