Utjecaj ekstrakcije potpomognute mikrovalovima na izolaciju polifenola kadulje (Salvia officinalis L.)

In order to determine the optimal conditions for the extraction of sage polyphenols, the influence of extraction solvent (water, 30 % (by volume) aqueous ethanol and 30 % (by volume) aqueous acetone), extraction time (3, 5, 7, 9 and 11 min) and microwave power (500, 600 and 700 W) on the composition and concentration of phenolic compounds of dry wild sage (Salvia officinalis L.) during microwave-assisted extraction (MAE) was studied. Optimized MAE method was compared with conventional extraction (CE). Based on the amount of total phenols, microwave power of 500 W and extraction time of 9 min were selected as optimal extraction conditions, resulting in higher content of polyphenols when compared with CE. Ethanol and acetone solutions were equally effective extraction solvents, both producing higher extraction capacity than water. Using HPLC coupled with UV/PDA, fourteen polyphenols were identified (caffeic and rosmarinic acid derivatives, luteolin- and apigenin-glycosides) with rosmarinic acid and luteolin glycosides at the highest concentrations. The mass fractions of all individual polyphenols were higher in the MAE extracts than in the CE ones.Da bi se odredili optimalni uvjeti ekstrakcije polifenola iz kadulje, ispitan je utjecaj otapala (voda, 30 %-tna vodena otopina etanola i 30 %-tna vodena otopina acetona), vremena ekstrakcije (3, 5, 7, 9 i 11 min) i snage mikrovalova (500, 600 i 700 W) na sastav i koncentraciju fenolnih spojeva tijekom ekstrakcije potpomognute mikrovalovima (MAE). Optimirana MAE metoda je zatim uspoređena s konvencionalnom metodom ekstrakcije (CE). Na temelju količine ukupnih fenola odabrani su optimalni uvjeti snage mikrovalova od 500 W i vremena ekstrakcije od 9 min pri kojima je ekstrahirana veća količina fenolnih spojeva u usporedbi s CE. Etanol i aceton bili su podjednako učinkovita otapala s većom djelotvornošću ekstrakcije od vode. Primjenom HPLC UV/PDA identificirano je četrnaest fenolnih spojeva (derivati kafeinske i ružmarinske kiseline, glikozidi luteolina i apigenina) i to najveći udjeli ružmarinske kiseline i glikozida luteolina. Svih pojedinačnih fenolnih spojeva bilo je više u ekstraktima dobivenim MAE metodom

Impact of stem cell source on allogeneic stem cell transplantation outcome in hematological malignancies

Background/Aim. Peripheral blood (PB) is used more frequently as a source of stem cells (SCs) for allogeneic transplantation. However, the influence of cell source on the clinical outcome of SC transplantation is not yet well established. The aim of this study was to compare the results of PBSC transplantation (PBSCT) with bone marrow transplantation (BMT) on the basis of engraftment, frequency and severity of immediate (mucositis, acute Graft versus Host Disease - aGvHD) and delayed (chronic GvHD - cGvHD) complications, as well as transplant-related mortality (TRM), transfusion needs, relapses and overall survival (OS). Methods. We analyzed 158 patients, women/men ratio 64/94 median age 29 (range 9-57), who underwent allogeneic SC transplantation between 1989 and 2009. All included patients had diseases as follows: acute myeloid leukemia (AML) - 39, acute lymphoblastic leukemia (ALL) - 47, chronic myeloid leukemia (CML) - 32, myelodysplastic syndrome (MDS) - 10, Hodgkin’s lymphoma (HL) - 2, multiple myeloma (MM) - 3, granulocytic sarcoma (GrSa) - 3, severe aplastic anemia (sAA) - 22. The patients underwent transplantations were divided into two groups: BMT group (74 patients) and PBSCT group (84 patients). Each recipient had HLA identical sibling donor. SCs from bone marrow were collected by multiple aspirations of iliac bone and from PB by one “Large Volume Leukapheresis” (after recombinant human granulocyte colony stimulating factor, rHuG-CSF) application (5-12 μg/kgbm, 5 days). Conditioning regimens were applied according to primary disease, GvHD prophylaxis consisted of combination of a cyclosporine A and methotrexate. Results. Engraftment, according to the count of polymorphonuclear and platelets, were significantly (p < 0.001) faster in the PBSCT vs BMT group. The needs for transfusion support were significantly (p < 0.01) higher in the BMT group. Those patients had more frequently oropharingeal mucositis grade 3/4 (33.3% vs 10.0%, p < 0.05). There were no significant differences in the incidence of aGvHD and cGvHD between the two groups. The patients who underwent PBSCT had more frequently extensive cGvHD in comparison with the BMT group (29.1% vs 11.29%, p < 0.05). SC source (SCS) had no significant influence on the TRM (21.62% vs 23.8%, p = 0.64) and the incidence of relapses (21.6% vs 29.7%, p = 0.32). Finally, the patients treated by BMT had a significantly better OS (logrank 2.33, p < 0.05). Conclusion. SCs harvesting from PB resulted in improved cell yield, faster engraftment, as well as in a decrease of immediate transplantation related complications with a reduced treatment cost. Allogeneic PBSCT were associated with more frequent extensive cGvHD, while the influence of SCS in TRM and relapses was not observed. Finally, the longterm OS was better in the patients treated by BMT. To verify impact of SC source on transplantation (PBSCT vs BMT) overall efficacy, more larger randomized clinical studies are needed

Validacija sprske verzije upitnika za procenu hronične opstruktivne bolesti pluća

Uvod/Cilj. Upitnik za procenu hronične opstruktivne bolesti pluća (HOBP) (engl. COPD Assessment Test - CAT) je jednostavan i pouzdan test namenjen za merenje ukupnog zdravstvenog stanja bolesnika sa HOBP i koristan je za upotrebu u svakodnevnoj kliničkoj praksi. Cilj ovog istraživanja bio je da se proceni validnost i opravdanost primene srpske verzije CAT. Metode. U studiji je učestvovalo 140 bolesnika u stabilnom stanju HOBP, ispitivanih u ambulantnim uslovima na Klinici za pulmologiju, Kliničkog centra Srbije u Beogradu i Institutu za plućne bolesti Vojvodine u Sremskoj Kamenici. Tokom prvog pregleda bolesnicima je učinjeno ispitivanje plućne funkcije (spirometrija), popunili su CAT upitnik i mMMR (modified Medical Research Council) skalu za procenu stepena dispneje. Pouzdanost CAT test-retesta je ispitivana kod 20 bolesnika od strane istog istraživača. Rezultati. Pokazali smo da srpska verzija CAT ima visoku internu konzistentnost sa Cronbach-ovim alfa 0.88. Test-retest analiza pokazala je dobru korelaciju između CAT rezultata u dve vremenske tačke (Spearmanov r = 0,681; p < 0,01). CAT je umereno korelirao sa mMRC skalom (r = + 0,57), blago sa forsiranim ekspiratornim volumenom u prvoj sekundi (FEV1), (r -0,214), uz pozitivnu korelaciju sa ukupnim brojem pogoršanja HOBP, ali bez jasne regularnosti sa promenom GOLD (Global Initiative for Chronic Obstructive Lung Disease) stadijuma. Zaključak. Srpska verzija CAT je pokazala visoku internu konzistentnost i test-retest pouzdanost. Ona predstavlja pouzdano, jednostavno i lako sredstvo za upotrebu koje se može koristiti u svakodnevnoj kliničkoj praksi za procenu zdravstvenog stanja kod bolesnika sa HOBP u Srbiji.Background/Aim. The Chronic obstructive pulmonary disease (COPD) Assessment Test (CAT) is a simple and reliable tool designed to measure overall COPD related health status and complement physician assessment in rou-tine clinical practice. Objective of this study was to evaluate the validity of the Serbian version of CAT. Methods. Study included 140 outpatients in the stable COPD, recruited from two centres: Clinic for Pulmonology, Clinical Center of Serbia, Belgrade, and Institute for Pulmonary Diseases of Vojvodina, Sremska Kamenica. All patients completed pulmonary function testing – spirometry, the CAT and the modified Medical Research Council (mMRC) dyspnea scale at baseline visit. The CAT test-retest reliability was tested in 20 patients by the same investigator (physician). Results.We demonstrated that Serbian version of CAT had high internal consistency with Cronbach’s alpha 0.88. Test-retest analysis showed good correlation between CAT scores in two time points (Spearman’s ρ = 0.681, p < 0.01). In our study the CAT correlated moderately to mMRC scale (ρ = +0.57), weakly to FEV1 (ρ -0.214), was positively related to number of exacerbations, but did not showed exact regularity with change in the Global Initiative for Chronic Obstructive lung disease (GOLD) stage. Conclusion.The Serbian version of CAT is a reliable, simple and easy-to-use tool that can be used in everyday clinical practice to assess the health status of COPD patients in Serbia

Mogući utjecaji rata u Hrvatskoj (1990.-1995.) na epidemiologiju raka štitne žlijezde

A Thyroid Cancer Registry containing data of patients treated at our department during a 20-year period (1980-1999) has been established. The aim of the study was to analyze the possible effects of the 1990-1995 war in Croatia on the prevalence, type, and age and sex distribution of thyroid carcinoma. Three different 2-year periods were analyzed: distant prewar (1980-1981), immediate prewar (1989-1990) and postwar (1998-1999) period. There was no statistically significant difference in age at diagnosis and sex ratio among the three study periods. The incidence of follicular cancer was at the upper limit for countries with normal iodine uptake when the 20-year period was analyzed, however, in the 1980-1981 period follicular cancer accounted for 34.9%, and in 1998-99 for only 2.8% of all cases. Papillary cancer was diagnosed in less advanced stages than others. Five-year survival for papillary, follicular and anaplastic cancer was 100%, 100% and 0%, respectively. Analysis of the 2-year periods for all cancer types except anaplastic cancer showed the mean age at onset to be on a decrease. In the 1998-1999 period, papillary cancer was diagnosed in a more advanced stage. The number of follicular cancer cases decreased from 22 in 1980-1981 to 3 in 19891990 and 2 in 1998-1999. Analysis of the epidemiology of thyroid cancer in the pre- and post-Chernobyl period did not reveal any increase in the number of papillary cancer in younger patients. It was concluded that in the postwar period, patients presented in more advanced stages of the disease. However, the effects of war on the epidemiology of thyroid carcinoma and other malignant tumors should be further investigated.Na našoj Klinici uspostavljeni Registar raka štitne žlijezde sadrži podatke o svim bolesnicima obrađenim u Klinici u 20godišnjem razdoblju (1980.-1999.). Cilj ovoga istraživanja bio je utvrditi moguće učinke rata u Hrvatskoj na učestalost, histološki tip te spolnu i dobnu raspodjelu bolesnika sa zloćudnim tumorom štitne žlijezde. Analizom su obuhvaćena 3dvogodišnja razdoblja uključujući vrijeme znatno prije rata (1980.-1981.), neposredno prije rata (1989.-1990.) i nakon rata(1998.-1999.). Nije utvrđena statistički značajna razlika u starosti i spolnoj raspodjeli tumora između tri navedena razdoblja. Učestalost folikularnog raka bila je na gornjoj granici učestalosti za zemlje bez gušavosti, s tim da je u razdoblju 1980.-1981. bilo ukupno 34,9%, a u razdoblju 1998.-1999. samo 2,8% folikularnog karcinoma. Petogodišnje preživljenje bilo je 100%, 100% odnosno 0% za papilarni, folikularni i anaplastični rak. U uspoređivanim razdobljima prosječna starost bolesnika se snižavala, ali se rak dijagnosticirao u sve uznapredovalijem stadiju. Analiza populacije prije i nakon Černobila nije pokazala povećan broj mladih bolesnika s papilarnim karcinomom. Na temelju rezultata ovoga istraživanja zaključeno je da nije bilo povećanja učestalosti karcinoma štitnjače u poratnom razdoblju, međutim, bolesnici su se javljali s uznapredovalijim stadijima tumora. Ovaj i moguće druge učinke rata na karcinom štitnjače, kao i na druge zloćudne tumore, valja dalje istraživati

The Influence of Microwave-Assisted Extraction on the Isolation of Sage (Salvia officinalis L.) Polyphenols

In order to determine the optimal conditions for the extraction of sage polyphenols, the influence of extraction solvent (water, 30 % (by volume) aqueous ethanol and 30 % (by volume) aqueous acetone), extraction time (3, 5, 7, 9 and 11 min) and microwave power (500, 600 and 700 W) on the composition and concentration of phenolic compounds of dry wild sage (Salvia officinalis L.) during microwave-assisted extraction (MAE) was studied. Optimized MAE method was compared with conventional extraction (CE). Based on the amount of total phenols, microwave power of 500 W and extraction time of 9 min were selected as optimal extraction conditions, resulting in higher content of polyphenols when compared with CE. Ethanol and acetone solutions were equally effective extraction solvents, both producing higher extraction capacity than water. Using HPLC coupled with UV/PDA, fourteen polyphenols were identified (caffeic and rosmarinic acid derivatives, luteolin- and apigenin-glycosides) with rosmarinic acid and luteolin glycosides at the highest concentrations. The mass fractions of all individual polyphenols were higher in the MAE extracts than in the CE ones

Acquired factor V inhibitors in a polytraumatized patient

Background. Factor V (FV) inhibitors are a rare disorder reported for the first time about fifty years ago, mostly with the unknown cause. The appearance of FV inhibitors is usually preceded by surgery, infections, administration of antibiotics or transfusions. Clinical manifestations of the presence of FV inhibitors vary from mild to severe and in some instances fatal hemorrhage. Case report. A healthy 51-year-old man with severe multiple injuries (traffic accident), and hemorrhage, which occurred during the orthopedic treatment, was admitted with hemoptysis, epistaxis and hematoma of the right upper leg, and with prolonged prothrombin time (PT), and activated partial thromboplastin time (aPTT). Treatment with vitamin K, fresh-frozen plasma and cryoprecipitate stopped the hemorrhage, but the results of coagulation tests were not normalized. The correction of aPTT and PT with normal plasma showed the decreased activity of FV (1%) due to the presence of inhibitors (titer 17.5 IU). The abnormal results of coagulation tests remained for three weeks, but without clinically manifested hemorrhagic syndrome. At the fourth week after the appearance of FV inhibitors PT, aPTT and the activity of FV became normal and antibodies disappeared spontaneously. Conclusion. Our patient with polytrauma developed a mild hemorrhagic syndrome due to the presence of FV inhibitors five weeks after the accident. Hemorrhage was treated with substitution therapy. The cause of the development of FV inhibitors was unclear (“fibrin glue” was not used during the orthopedic treatment). Factor V inhibitors disappeared spontaneously within four weeks. The fast spontaneous disappearance of FV inhibitors in our patient, confirmed the observations of some authors that they disappeared faster in those patients who were surgically treated prior to their appearance

Autologous hematopoietic stem cell transplantation in combination with immunoablative protocol in secondary progressive multiple sclerosis: A 10-year follow-up of the first transplanted patient

Introduction. Multiple sclerosis (MS) is an immunemediated disease of the central nervous system that affects young individuals and leads to severe disability. High dose immunoablation followed by autologous hemopoietic stem cell transplantation (AHSCT) has been considered in the last 15 years as potentialy effective therapeutic approach for agressive MS. The most recent long-time follow-up results suggest that AHSCT is not only effective for highly aggressive MS, but for relapsing-remitting MS as well, providing long-term remission, or maybe even cure. We presented a 10- year follow-up of the first MS patient being treated by immunoablation therapy and AHSCT. Case report. A 27-year-old male experienced the first symptoms - intermitent numbness and paresthesia of arms and legs of what was treated for two years by psychiatrist as anxiety disorder. After he developed severe paraparesis he was admitted to the Neurology Clinic and diagnosed with MS. Our patient developed aggressive MS with frequent relapses, rapid disability progression and transition to secondary progressive form 6 years after MS onset [the Expanded Disability Status Scale (EDSS) 7.0 Ambulation Index (AI) 7]. AHSCT was performed, cyclophosphamide was used for hemopoietic stem cell mobilization and the BEAM protocol was used as conditionig regimen. No major adverse events followed the AHSCT. Neurological impairment improved, EDSS 6.5, AI 6 and during a 10-year followup remained unchanged. Brain MRI follow-up showed the absence of gadolinium enhancing lesions and a mild progression of brain atrophy. Conclusion. The patient with rapidly evolving, aggressive, noninflammatory MS initialy improved and remained stable, without disability progression for 10 years, after AHSCT. This kind of treatment should be considered in aggressive MS, or in disease modifying treatment nonresponsive MS patients, since appropriately timed AHSCT treatment may not only prevent disability progression but reduce the achieved level of disability, as well

Autologous stem cell transplantation in the treatment of Hodgkin's disease

Background/Aim. High-dose chemotherapy with autologous stem cell transplantacion (ASCT) has shown to produce long-term disease-free survival in patients with chemotherapysensitive Hodgkin disease. The aim of the study was to evaluate efficacy of ASCT in the treatment of Hodgkin's disease. Methods. Between May 1997 and September 2008, 34 patients with Hodgkin's disease in median age of 25 (range 16-60) years, underwent ASCT. Autologous SCT were performed as consolidation therapy in one poor-risk patients with complete response (CR) and in 10 patients in partial remission (PR) after induction chemotherapy (32.5%), for chemosensitive relapse (CSR 1 and CSR 2) in 47% patients and in 20.5% patients with chemoresistant disease (CRD). All except one patient were in stage III/IV, extranodal site of disease had 24 patients and bulky disease had l0 patients. All the patients received a uniform preparatory regimen (BEAM). Results. An overall response was achieved in 30 of 32 evaluated patients, with 62.5% in CR and 31.25% in PR. After applying radiotherapy, two patients with PR after ASCT reached CR. Median follow-up was 15.5 months (range 3-133 months). The probability of overall survival (OS) and progression-free survival (PFS) at a 3-year period for all patients was 51.9 % and 48.9%, respectively. For 22 patients in CR after ASCT, a 3-year DFS was 66.5%. Estimates of 2.5-year survival were 14.3%, 61.9% and 100% for CRD, CSR and for patients with CR/PR, respectively (p &lt; 0.01). However, when patients undergoing consolidation were analyzed separately from those in CSR, no significant difference in OS and PFS was observed according to the disease status at ASCT. In univariate analysis for OS, PFS i DFS, extranodal site of disease and disease bulk had no predictive value. Twelve patients died. The main cause of death was Hodgkin's disease. Transplant-related mortality was 3.1%. One patient with CRD developed secondary acute myeloid leukemia and died 28 months after the transplantation. Conclusion. Autologous SCT is efficient as consolidation therapy in high-risk patients and in chemosensitive relapse, but it has no benefit in patients with chemoresistant disease