Molecular basis of genetic rearrangements induced by IS5 elements in polA1 mutator strain of E.coli K12

U našem prethodnom radu na polA1 mutantima pokazali smo da polA1 kreira mutacije isključivo tipa minus fs i delecija. U mutacionom spektru histidinskih auksotrofa bila je prisutna i jedna specifična kategorija delecija, velike delecije, koje su prolazile preko celog his operona. Veličina delecija je ukazivala da one nastaju u procesu rekombinacije izmedju homologih sekvenci, a poznato je da u polA mutantima dolazi do povećane frekvence rekombinacije. Ovaj rad predstavlja završni deo naše originalne studije polA1 indukovane mutageneze praćene na histidinskom operonu hromozoma E.coli. Analizirani su molekularni mehanizmi nastajanja velikih delecija, ili proces rekombinacije koji promoviše ove genomske rearanžmane. Pokazali smo da velike delecije nastaju kao rezultat rekombinacije izmedju dve IS5 sekvence koje se nalaze sa obe strane his operona. Sve četiri nezavisno izolovane delecije su bile jednake i iznosile su 35 kb. Analizom sekvence rekombinovanih IS5 elemenata pokazali smo da rekombinacija uvek prolazi kroz levi terminus IS5 i da se odigrava po tipu “site specific” rekombinacije. Levi terminus IS5 elementa ima karakteristično zakrivljenje DNK uslovljeno njenom primarnom strukturom i sadrži mesto za vezivanje IHF proteina. Rezultati ovog rada zajedno sa podacima iz literature ukazuju na moguću ulogu IHF proteina u nastajanju velikih delecija i u procesu rekombinacije. Primenom PCR tehnologije pokazli smo da je frekvenca nastajanja velikih delecija povećana kod polA1 i dam mutanata i da velike delecije nastaju na recA nezavisan način. PolA1 ne pokazuje nikakvu specifičnost jer se rekombinacija dešava na isti način i kod divljeg soja, ali je frekvenca rekombinacije dva puta povećana u polA1 mutantima u odnosu na divlji soj.In our previous work on polA1 mutants we have shown that polA1 creates exclusively minus fs and deletions types. In mutational spectra of histidine auxotrophs, one specific category of deletions, large deletions, that cover all his operon, was present. The size of deletions pointed out that they appear in a process of recombination between homology sequences; it is known that polA1 mutants show an increased frequency of recombination. This work presents final part of our original study of polA1-induced mutagenesis followed on histidine operon of chromosome E.coli. Molecular mechanisms of appearances of large deletions, or process of recombination promoting this chromosomal rearrangement, was analysed. We showed that large deletions appear as a result of recombination between two IS5 sequences located on both sides of his operon. Four independently isolated deletions were identical, 35 kb long. Sequence analysis of recombinant IS5 elements was show that recombination always goes throught the left terminus of IS5, as a “site specific” recombination. Left terminus of IS5 contains a characteristic sequence-directed bent DNA, including a binding site for IHF protein. The data from literature, including the present results, point to a possible role of IHF protein in appearance of large deletions and process of recombination. Applying a PCR technology, we show that the frequency of large deletions is increased in polA1 and dam mutants, and that large deletions appear on recA independent way. PolA1 is not sequence specific because the process of recombination between two IS5 is identical in wild type and polA mutants, but the frequency of recombination is two times higher in regard to wild type

Cost Management at Higher Education Institutions – Cases of Bosnia and Herzegovina, Croatia and Slovenia

Higher education expenditures persistently rise due to various economic, demographic and socio-cultural reasons. This caused repeated calls for reforms of the economic model in the higher education sector and dramatically increased the importance of the economic evaluations in the last decades due to concerns for efficiency. The above academic challenges led us to pioneering an attempt to evaluate the capabilities of financial management tools for three Western Balkan countries, i.e. Bosnia and Herzegovina, Croatia and Slovenia. The precondition for successful reform processes is certainly a comprehensive and high-quality accounting information system that meets not only the requirements of external reporting but also the requirements of internal users, especially the management of HEIs. In that context, the main aim of this paper is to overview the legal and organizational accounting systems' characteristics focusing on external and internal reporting requirements, and study the level of development and usage of cost accounting at HEIs in selected countries. Therefore, our paper employs research methodology based on the survey conducted. The results show great differences in legal and organizational characteristics of accounting systems among the countries as well as in the development stages of cost accounting systems, which mainly focus on inconsistent overhead allocation as well as different accounting basis usage. The research results confirm poorly conceptualised and structured reporting of accounting information for management purposes, offering several applicable platforms for creation of performance management approaches and strategies in the public sector

Ekonomska obilježja proizvodnje mlijeka u Sarajevsko-Romanijskoj regiji

Sarajevo-Romanija region entirely belongs to the hilly and mountainous region where livestock production is the dominant form of agriculture. The paper presents results of a research conducted on a sample of ten farms engaged in dairy production. The use of scientific methods has helped us gain a clearer picture of basic organizational and economic characteristics of dairy production in the Sarajevo-Romanija region. Farm production conditions are characterized by marked heterogeneity, resulting in variability of economic performance. In cases where there was a higher degree of fulfillment of prescribed organizational and technological measures, satisfactory economic performance was achieved and the farms, with additional improvements implemented, have further prospects in milk and calf production.Sarajevsko-romanijska regija u potpunosti pripada brdsko-planinskom području, gdje je stočarstvo preovlađujući oblik poljoprivredne proizvodnje. U radu su predstavljeni rezultati istraživanja sprovedenih na uzorku od deset porodičnih gazdinstava koja se bave proizvodnjom mlijeka. Uslovi proizvodnje na gazdinstvima karakterišu se izraženom heterogenošću, što prouzrokuje varijabilitet ekonomskog uspjeha poslovanja. U slučajevima gdje je zabilježen viši stepen ispunjenja propisanih organizaciono-tehnološkim mjera ostvareni su zadovoljavajući ekonomski rezultati i ta gazdinstva, uz dodatna poboljšanja, imaju dalju perspektivu u proizvodnji mlijeka i teladi

Efficacy and Safety of Ombitasvir/Paritaprevir/Ritonavir ± Dasabuvir Regimen in Haemodialysis Patients With Hepatitis C Virus Infection: A Case Series

Uvod: Infekcija virusom hepatitisa C (HCV) česta je među bolesnicima na hemodijalizi (HD) i važan je uzrok morbiditeta i mortaliteta. Kod bolesnika s kroničnom bolesti bubrega (CKD), rizici za negativne ishode značajno su veći u pacijenata zaraženih HCV-om nego u onih bez HCV infekcije, uključujući napredovanje u cirozu, hepatocelularni karcinom i smrtnost povezanu s jetrom. Ombitasvir (OBV), paritaprevir (PTV), ritonavir (r) i dasabuvir (DSV) metaboliziraju se u jetri i stoga ne zahtijevaju prilagodbu doze u bolesnika s bilo kojim stupnjem oštećenja bubrega. Ciljevi: Proučavali smo sigurnost i djelotvornost OBV/PTV/r+DSV u maloj skupini pacijenata zaraženih HCV-om na terapiji hemodijalizom. Metode: Analizirali smo liječenje ombitasvir / paritaprevir / ritonavir i dasabuvir; (3-DAA režim OBV / PTV / r + DSV ± RBV). Primarna završna točka bila je održivi virološki odgovor 12 tjedana nakon liječenja (SVR12). Prikupili smo podatke o nuspojavama (AEs), ozbiljnim neželjenim učincima i abnormalnostima laboratorija. Rezultati: Od 7 liječenih bolesnika, 6 su bili muški i 1 ženski, a svi su bili zaraženi genotipom 1 (5 GT1b, 2 GT1a). Jedan pacijent imao je kompenziranu cirozu jetre, a šest bolesnika nije imalo cirozu jetre te niti jedan nije bio primatelj transplantata jetre. Svih sedam bolesnika završilo je 12 tjedana liječenja i postiglo SVR12. Istodobni lijekovi morali su se mijenjati s početkom liječenja kod 5 od 7 bolesnika. Jedan od bolesnika imao je značajno smanjenje razine hemoglobina, broja bijelih krvnih stanica i trombocita tijekom razdoblja liječenja. Najčešće nuspojave bile su mučnina, proljev. Nuspojave su primarno bile blage, a nijedan pacijent nije prekinuo liječenje zbog AE. Zaključak: Liječenje OBV / PTV / r + DSV ± RBV bilo je dobro podnošljivo i rezultiralo je visokim stopama SVR12 (100%) za bolesnike s HCV GT1b/1a na hemodijalizi.Background: Hepatitis C virus (HCV) infection is common among patients on haemodialysis (HD) therapy and is an important cause of morbidity and mortality. In patients with chronic kidney disease (CKD), the risks for negative outcomes are significantly higher in HCV-infected patients than in those without HCV infection, including progression to cirrhosis, hepatocellular carcinoma and liver-related mortality. Ombitasvir (OBV), paritaprevir (PTV), ritonavir (r), and dasabuvir (DSV) are all hepatically metabolized and, therefore, require no dose adjustment in patients with any degree of renal impairment. Aims: We studied the safety and efficacy of OBV/PTV/r + DSV in a small group of HCV infected patients on haemodialysis therapy. Methods: Treatment course with ombitasvir/paritaprevir/ritonavir and dasabuvir; (3-DAA regimen of OBV/PTV/r+DSV±RBV) was analysed. Pre-treatment evaluation of HCV infection included HCV RNA, genotype, and liver fibrosis assed by transient fibroelastography (FibroScan). The stage 5 CKD was defined as an eGFR of <15 mL/min/1.73 m2, respectively; those on haemodialysis were considered to have stage 5 CKD or end-stage renal disease (ESRD). Demographic data and concomitant medication were retrieved from patients’ records. The primary endpoint was sustained virologic response at post-treatment week 12 (SVR12). We collected data on on-treatment adverse events (AEs), serious AEs, and laboratory abnormalities. Results: Among 7 treated patients, 6 were male and 1 female, all were infected with genotype 1 (5 GT1b, 2 GT1a). Patient had compensated liver cirrhosis and six patients did not have liver cirrhosis, none were liver transplant recipients. All of seven patients completed 12 weeks of treatment and achieved SVR12. Concomitant medication had to be modified with the treatment initiation in 5 out of 7 patients. One of the patients presented with a significant decrease in haemoglobin level, white blood cell and platelet count during the treatment period. The most frequent adverse events were nausea, diarrhoea. Adverse events were primarily mild, and no patient discontinued treatment due to an AE. Conclusions: Treatment with OBV/PTV/r +DSV ± RBV was well-tolerated and resulted in high rates of SVR12 (100%) for patients with HCV GT1b/1a on haemodialysis

Craniocervical Dystonia Questionnaire (CDQ-24): Validation and Cross-Cultural Adaptation in Serbian Patients

The purpose of this study was to investigate the validation of the translated and culturally adapted CDQ-24 questionnaire on a group of Serbian patients. The study was comprised of 100 consecutive patients with idiopathic cervical dystonia (CD) and blepharospasm (BSP) who were evaluated at the Institute of Neurology, Clinical Centre of Serbia in Belgrade between March and June 2007. The linguistic validation of CDQ-24 involved 3 steps, according to an internationally accepted methodology. Most of the patients with CD and BSP accepted the CDQ-24 questionnaire. The internal consistency reliability ranged from 0.81 to 0.97. The mean total score of the CDQ-24 was 35.6±23.5. Patients with BSP had better HRQoL scores in the Pain subscale (p=0.025) compared with CD patients. However, patients with CD had better HRQoL sores in the Activities of Daily Living subscale (p=0.028) compared with BSP patients. Statistically significant positive correlations were registered between the Dystonia Movement Scale score and almost all CDQ-24 scales. The Serbian version of CDQ-24 should be recommended for HRQoL evaluation among patients with CD and BSP as an important outcome measure

Clinical characteristics of multiple system atrophy in Serbian population

Background/Aim. Mulstiple system atrophy (MSA) is a neurodegenerative central nervous system disorder, characterized by any combination of extrapyramidal, cerebellar, pyramidal or autonomic disturbance. The aims of our study were to define clinical characteristics of MSA patients in our population, to account for neuroradiological and electrophysiological profile of the disease and to evaluate one-dose levodopa response. Methods. We have diagnosed 29 patients as MSA, with disease duration from the first symptom 5 years on average on examination. The examinating procedure included an anamnesis and complete neurological investigations, as well as neurootological and neuroradiological examinations. The study included the patients of the Institute of Neurology of the Clinical Center of Serbia in the period of 1996-2001, who completed both clinical and diagnostic criteria for a possible and probable MSA. Results. Autonomic disturbances were documented in 93.1%, whereas extrapyramidal symptoms were seen in additional 89.3% with symmetrical onset in 60%. Levodopa response was poor or moderate. Cerebellar signs were present in 63%, while pyramidal signs occured in 78.7%. There was no a cognitive deterioration (MMSE &gt; 24). CT scan and MRI showed cerebellar and brainstem atrophy, as well as diffuse cortical atrophy. Conclusion. Failure of additional diagnostic procedures to distinguish MSA patients required a precise understanding of their clinical specificities. Our results support this statement

When do the symptoms of autonomic nervous system malfunction appear in patients with Parkinson’s disease?

Background/Aim. Dysautonomia appears in almost all patients with Parkinson’s disease (PD) in a certain stage of their condition. The aim of our study was to detect the development and type of autonomic disorders, find out the factors affecting their manifestation by analyzing the potential association with demographic variables related to clinical presentation, as well as the symptoms of the disease in a PD patient cohort. Methods. The patients with PD treated at the Clinic of Neurology in Belgrade during a 2-year period, divided into 3 groups were studied: 25 de novo patients, 25 patients already treated and had no long-term levodopa therapy-related complications and 22 patients treated with levodopa who manifested levodopa-induced motor complications. Simultaneously, 35 healthy control subjects, matched by age and sex, were also analyzed. Results. Autonomic nervous system malfunction was defined by Ewing diagnostic criteria. The tests, indicators of sympathetic and parasympathetic nervous systems, were significantly different in the PD patients as compared with the controls, suggesting the failure of both systems. However, it was shown, in the selected groups of patients, that the malfunction of both systems was present in two treated groups of PD patients, while de novo group manifested only sympathetic dysfunction. For this reason, the complete autonomic neuropathy was diagnosed only in the treated PD patients, while de novo patients were defined as those with the isolated sympathetic dysfunction. The patients with the complete autonomic neuropathy differed from the subjects without such neuropathy in higher cumulative and motor unified Parkinson’s disease rating score (UPDRS) (p < 0.01), activities of daily living scores (p < 0.05), Schwab-England scale (p < 0.001) and Hoehn-Yahr scale. There was no difference between the patients in other clinical-demographic characteristics (sex, age at the time of diagnosis, actual age, duration of disease, involved side of the body, pain and freezing), but mini mental status (MMS) score and Hamilton depression and anxiety rating scale were significantly lower (p < 0.05). Conclusion. Our results confirm a high prevalence of autonomic nervous system disturbances among PD patients from the near onset of disease, with a predominant sympathetic nervous system involvement. The patients who developed complete autonomic neuropathy (both sympathetic and parasympathetic) were individuals with considerable level of functional failure, more severe clinical presentation and the existing anxiety and depression. [Projekat Ministarstva nauke Republike Srbije, br. 175090

JAK2V617F mutation in a patient with B-cell chronic lymphocytic leukemia and prefibrotic primary myelofibrosis

Uvod Sekundarni maligniteti, naročito solidni tumori, česti su kod bolesnika s hroničnom limfocitnom leukemijom (HLL), ali retko se sreće udruženost mijeloproliferativnih neoplazmi i HLL. Prikaz bolesnika Prikazujemo muškarca starog 67 godina sa B ćelijskom HLL kod koga se nakon devet godina razvila primarna mijelofibroza (PMF). Bolesnik je lečen alkilišućim agensima i analozima purina, što može biti predisponirajući faktor za razvoj mijeloproliferativnog oboljenja. JAK2V617F mutacija nije otkrivena prilikom postavljanja dijagnoze HLL, ali je utvrđena posle devet godina, kada se razvila PMF, što ukazuje na to da su B ćelijska HLL i PMF neoplazme koje potiču od različitih ćelijskih klonova. Zaključak Patogenetski mehanizmi udruženosti mijeloproliferativne i limfoproliferativne neoplazme kod bolesnika nisu razjašnjeni. Potrebna su dalja istraživanja radi utvrđivanja da li ove maligne bolesti potiču od dva različita ćelijska klona ili nastaju od iste pluripotentne matične ćelije hematopoeze.Introduction Secondary malignancies, particularly solid tumors, are common in patients with chronic lymphocytic leukemia (CLL), but association of myeloproliferative neoplasms and chronic lymphocytic leukemia in the same patient is very rare. Case Outline We report of a 67-year-old man with B-cell chronic lymphoid leukemia (B-CLL) who developed primary myelofibrosis (PMF) nine years after initial diagnosis. Patient received alkylation agents and purine analogue, which can be a predisposing factor for the development of myeloproliferative neoplasms. JAK2V617F mutation was not present initially at the time of CLL diagnosis, but was found after nine years when PMF occurred, which indicates that B-CLL and PMF represent two separate clonal origin neoplasms. Conclusion Pathogenic mechanisms for the development of myeloproliferative and lymphoproliferative neoplasms in the same patient are unknown. Further research is needed to determine whether these malignancies originate from two different cell clones or arise from the same pluripotent hematopoietic stem cell

The influence of the UV irradiation intensity on photocatalytic activity of ZnAl layered double hydroxides and derived mixed oxides

Layered double hydroxides (LDHs) have been studied to a great extent as environmental-friendly complex materials that can be used as photocatalysts or photocatalyst supports. ZnAl layered double hydroxides and their derived mixed oxides were chosen for the investigation of photocatalytic performances in correlation with the UV intensities measured in the South Pannonia region. Low supersaturation coprecipitation method was used for the ZnAl LDH synthesis. For the characterization of LDH and thermal treated samples powder X-ray diffraction (XRD), scanning electron microscopy (SEM), electron dispersive spectroscopy (EDS), nitrogen adsorption-desorption were used. The decomposition of azodye, methylene blue was chosen as photocatalytic test reaction. The study showed that the ZnAl mixed oxide obtained by thermal decomposition of ZnAl LDH has stable activity in the broader UV light irradiation range characterizing the selected region. Photocatalytic activity could be mainly attributed to the ZnO phase, detected both in LDH and thermally treated samples. The study showed that the ZnAl mixed oxide obtained by the calcination of ZnAl LDH has a stable activity within the measured UV light irradiation range; whereas the parent ZnAl LDH catalyst did not perform satisfactory when low UV irradiation intensity is implied

The role of FasR/FasL system in pathogenesis of myeloprolyferative neoplasms

Mijeloproliferativne neoplazije (MPN) su hematološki maligniteti koji se karakterišu nekontrolisanom ćelijskom proliferacijom i poremećajem u procesu apoptoze. Sistem FasR/FasL je uključen u kontrolu apoptoze u različitim tipovima ćelija. U ovom radu je izučavana uloga sistema FasR/FasL u patogenezi mijeloproliferativnih neoplazija. Upoređena je ekspresija FasR i FasL između pacijenata sa MPN (24) i zdravih kontrola korišćenjem metode 'real-time' PCR. Detektovana je povećana ekspresija FasR kod pacijenata sa MPN. Nije utvrđena razlika u ekspresiji FasL. Mutacija B617F u JAK2 genu, karakteristična za MPN, je nađena kod 13 od 24 pacijenta. Pokazano je da ekspresija FasR i FasL nije povezana sa prisustvom B617F JAK2 mutacije.Myeloproliferative neoplasms (MPN) are hematological malignancies characterized by uncontrolled cell proliferation and impaired apoptosis. The FasR/FasL system is involved in the control of apoptosis in different cell types. Here we have investigated the role of FasR/FasL in the pathogenesis of MPNs. We compared FasR/FasL expression between MPN patients (24) and healthy individuals using the real-time PCR assay. We found an increase of FasR expression in MPN patients. No difference was detected in FasL expression. Mutation V617F in the JAK2 gene, a hallmark of MPN, was detected in 13/24 patients. We found that neither FasR nor FasL expression were related to the presence of JAK2 V617F mutation