Novel Therapy Approaches in β-Thalassemia Syndromes — A Role of Genetic Modifiers

The β-thalassemia syndromes are heterogeneous autosomal recessive hereditary disorders, caused by alterations in the HBB gene and characterized by absent or reduced β-globin chain synthesis. The β-thalassemia phenotypes are variable, ranging from severe, transfusion-dependent thalassemia major to mild, asymptomatic thalassemia trait. This interpatient clinical variability has swayed researchers toward identifying genetic modifiers for these disorders. Primary modifiers refer to type of alterations affecting β-globin gene. Secondary modifiers include variations in genes affecting α/β-globin chain equilibrium, such as genes involved in the γ-globin gene expression and genes affecting the amount and stability of α-globin chains. Tertiary modifiers are gene variations affecting the phenotype with regard to the complications caused by β-thalassemia syndromes. A role of secondary genetic modifiers in ameliorating the clinical phenotype has been observed. Secondary genetic modifiers are the most common targets for modern therapy and could be located within α- and γ-globin genes or outside globin gene cluster. The most potent secondary modifier genes are γ-globin genes. Production of fetal hemoglobin (HbF) trough adulthood ameliorates the severity of β-thalassemia phenotype. Large family and genome-wide association studies have shown that regions outside of the β-globin gene cluster are also implicated in γ-globin gene expression regulation. HBS1-MYB intragenic region and BCL11A gene have been particularly studied. Variants within these loci, along with γ-globin gene variants, account for approximately 50% of the HbF level variation, suggesting that additional factors are involved (transcription regulators (KLF1), regulators of α-globin chain stability (AHSP), epigenetic regulators (FoP)). Until recently a definitive cure for β-thalassemia could be achieved with bone marrow transplantation. However, it is available for less than 30% of the patients and bears a significant risk of morbidity and mortality. Alternative strategies, such as gene therapy and development of induced pluripotent stem cells (iPSCs) have been explored. The targets for gene therapy are hematopoietic stem cells, which are harvested from patient bone marrow or peripheral blood, purified by immunoselection, transduced by “therapeutic gene” aimed at correcting the effect of defective β-globin gene, and returned to the patient. Various types of vectors have been considered for gene transfer, including non viral (tRNK and ribozymes) and viral (retroviral and lentiviral vectors). In the past few years, iPSCs emerged as an interesting candidate for gene transfer. The feature that makes these cells appealing in the field of gene therapy is their susceptibility to gene correction by homologous recombination. Therapy protocols based on molecular basis of β-thalassemia are the best example of novel approaches in disease treatment

Rapid Cytoreduction by Plateletapheresis in the Treatment of Thrombocythemia

The objective of this chapter is to provide a systematic overview of current knowledge regarding therapeutic apheresis—primarily therapeutic plateletapheresis (TP)—and to summarize evidence-based practical approaches related to cytapheresis treatment of “hyperthrombocytosis” or “extreme thrombocytosis” (ETC). Our results of platelet (Plt) quantitative/qualitative analyses and evaluation of efficacy of apheresis systems/devices—on the basis of Plt removal and in vivo Plt depletion—will be presented. Our preclinical researches confirmed that in Plt concentrates, the initial ratio of discoid shapes was 70%, spherical 20%, and less valuable (dendritic/balloonized) shapes 10%—with morphological score of platelets (MSP = 300–400). After storage, the ratio of discoid and spherical shapes was decreased, while the less valuable ones progressively increased (MSP = 200). Electron microscopy has shown discoid shapes with typical ultrastructural properties. Spherical shapes with reduced electron density and peripheral location of granules/organelles were detected. Also, dendritic shapes with cytoskeletal “rearrangement,” membrane system integrity damages, and pseudopodia formations were documented. Our clinical study demonstrated that TP was useful in ETC treatment and should help prevention of “thrombo-hemorrhagic” events—until chemotherapy, antiplatelet drugs, and other medication take effect. During TP treatment, Plt count and morphology/ultrastructure were examined. Plt functions by multiplate analyzer were evaluated. We concluded that intensive TP was an effective, safe, and rapid cytoreductive treatment for ET

Prisustvo cinka u mišićnom tkivu babuške i deverike u akumulacijama Gruža i Bovan

Zagađenje slatkovodnih ekosistema širokim spektrom zagađivača postalo je veoma važno pitanje u poslednjih nekoliko decenija, ne samo zbog uticaj na vodosnabdevanje, već i zbog štete koju nanosi vodenim organizmima. Smatra se da su metali najvažniji oblik zagađenja voda zbog toksičnosti, perzistentnosti i akumulacije u vodenim organizmima. Vodeni organizmi su u širokoj upotrebi biološkog monitoringa promena nivoa antropogenih zagađujućih materija. Poznavanje koncentracije metala u ribama je važno kako za pravilno planiranje upravljanjа vodama tako i u pogledu ljudske ishrane. Zn je esencijalni metal i kao takav igra važnu ulogu u biološkim sistemima, ali takođe prekomernim unosom može proizvesti toksičan efekat. Akumulacije Gruža i Bovan su formirane osamdesetih godina za vodosnabdevanje, ali su danas i značajne ribolovne vode. Antropogeni uticaj na ove akumulacije je vrlo izražen. Konstantno je direktno ispuštanje neprečišćenih otpadnih voda, okolno zemljište se intenzivno obrađuje uz primenu invazivnih agrotehničkih mera (prekomerna upotreba pesticida, herbicida i fungicida), ali i širenje vikend naselja koja uglavnom nemaju regulisano skladištenje i ispuštanje otpadnih voda. Babuška (Carassius gibelio) i deverika (Abramis brama) su prikupljene iz akumulacija Gruža i Bovan, a uzorci mišićnog tkiva su analizirani na prisustvo cinka (Zn) kako bismo utvrdili da li postoji razlika u akumulaciji ovog metala u mišićnom tkivu između ove dve vrste, kao i između iste vrste u dve različite akumulacije. Koncentracija Zn u mišićnom tkivu babuške je bila znatno veća nego u mićnom tkivu deverike u obe akumulacije, ali nije postojala bitna razlika u koncentracijama između dve akumulacije. Koncentracija cinka u mišićima obe vrste u obe akumulacije je na prihvatljivom nivou za ljudsku ishranu

Odnos autohtonih i alohtonih ribljih vrsta u nekim akumulacijama Srbije

Poslednjih godina u svetu je veoma aktuelno pitanje alohtonih ribljih vrsta. Bilo da su unete slučajno ili namerno, alohtone vrste najčešće uspevaju da opstanu u novom okruženju, u kojem zbog degradiranosti staništa i narušenih biocenoloških veza vremenom postaju dominantne. Alohtone vrste često imaju invazivan karakter jer se brzo i nekontrolisano šire u nove ekosisteme gde se ponašaju kao kompetitori za prostor i hranu autohtonim vrstama i polako ih potiskuju. Ovim radom prikazan je odnos u brojnosti i biomasi autohtonih i alohtonih ribljih vrsta u nekim akumulacijama u Srbiji. Tokom istraživanja obraćena je pažnja na veličinu i starost akumulacija u kojima je praćena ihtiocenoza. Istraživanje je obuhvatilo 15 akumulacija različite starosti, tipa postanka, morfometrijskih, fizičkih i hemijskih osobina. Konstatovano je prisustvo 20 vrsta riba iz 9 familija, sa izrazitom dominacijom porodice Cyprinidae. Od ovog broja 8 vrsta su alohtone i pripadaju familijama Salmonidae (Oncorhynchus mykiss), Cyprinidae (Carassius auratus, Arystichtys nobilis, Hypophthalmichthys molitrix, Ctenopharyngodon idella, Pseudorabora parva), Ictaluridae (Ictalurus nebulosus - cverglan) i Centrarchidae (Lepomis gibbosus - sunčica) (Simić i Simić, 2009). Cverglan i sunčica potiču iz Severne Amerike, dok su ostale azijske vrste. U najvećem broju akumulacija, po brojnosti i biomasi, zastupljenije su autohtone riblje vrste u odnosu na alohtone. Izuzetak je Oblačinsko jezero, koje pripada grupi malih i starih jezera, gde brojnošću dominiraju alohtone vrste sa 61,63%, kao i biomasom sa čak 80,18%. Velika zastupljenost biomase alohtonih ribljih vrsta je i u jezeru Buljanka, 53.43%, dok njihova brojnost iznosi 17.07%. Najveći udeo u biomasi ima sivi tolstolobik. Autohtona ihtiocenoza dominira u jezeru Ćelije i Zavojskom jezeru koja su u grupi starijih i većih jezera. U ovoj grupi su i akumulacije Gruža, Vlasina i Bovan, gde se zbog neplanskog poribljavanja brojnost alohtonih vrsta približava abundanci autohtonih. Sa druge strane, planskim poribljavanjem očuvana je autohtona ihtiofauna u Miloševom i Ribničkom jezeru, koja su male veličine i stara. Konstatovano je da u najvećem broju jezera i dalje dominiraju autohtone vrste u odnosu na alohtone. Uticaj alohtonih ribljih vrsta je uvek veći u manjim i starijim akumulacijama, upravo zbog većeg diverziteta autohtone zajednice u njima. Poznajući invazivni karakter i negativan efekat alohtonih vrsta na ekosisteme u koje su unete, kao i na faunu, neophodno je stalno pratiti stanje na terenu

Uzgoj linjaka (tinca tinca) u laboratorijskim uslovima

Istraživanja biodiverziteta kopnenih voda Srbije sa ciljem konzervacije u in situ i/ili ex situ uslovima sprovode se u proteklih 10 godina u okviru više istraživačkih projekata. Kao rezultat ovih istraživanja formirana je aplikaciono-informativna baza podataka pod nazivom: Biodiverzitet akvatičnih ekosistema Srbije-ex situ konzervacija „BAES ex situ”. Na osnovu podataka iz baze, „BAES ex situ”, sprovedena su istraživanja tokom 2008. godine sa ciljem izrade Programa za unapređenje ribarstva u slivu Velike, Zapadne i Južne Morave (i dela toka Dunava, Save, kanala hidrosistema DTD i voda Pančevačkog rita). Dobijeni podaci jasno ukazuju da je linjak (Tinca tinca) u odnosu na istraživanja od pre 10 i 20 godina znatno manje zastupljen u slivu Velike Morave. Svi nalazi linjaka južno od Save i Dunava tokom ovog istraživanja su iz hidroakumulacija Srbije (Vlasina, Ćelije) i uneseni su poribljavanjem iz uzgajališta ili prirodnih voda sa područja Vojvodine. Stanje populacija linjaka prema literaturnim podacima Kirchhofer & Hefti (1996), Kottelat & Freyhof (2007) ukazuju da bez obzira na njegovo široko geografsko rasprostranjenje svuda su u manjem ili većem opadanju, a kao osnovni uzrok se navodi degradacija i uništavanje vodenih staništa, pre svega onih koja su bogata makrovegetacijom. Konstatovano stanje populacija linjaka u vodenim ekosistemima Srbije nameće potrebu konzervacije, veštačkog mrešćenja i repopulacije ugroženih prirodnih populacija. Cilj rada jeste istražiti mogućnosti uzgoja linjaka u laboratorijskim uslovima, u pogledu različite ishrane, u cilju uspešne konzervacije. Istraživanje je sprovedeno u Akvarijumu ″Kragujevac″, koji je deo Instituta za biologiju i ekologiju Prirodno-matematičkog fakulteta u Kragujevcu. Eksperiment je trajao 11 meseci (januar-novembar 2010.). Nasadni materijal činile su jedinke donete iz ribnjaka ″Mošorin″, uzrasne kategorije 0+. Formirane su tri eksperimentalne grupe sa po tri akvarijuma, dimenzija 100 x 40 x 40 cm, zapremine 160 l. Ukupan broj nasađene riblje mlađi iznosio je 30 individua (≈160 g) po grupi, odnosno 10 jedinki po akvarijumu, prosečne telesne mase 5,52 g. Ishrana je vršena na sledeći način: prva grupa hranjena je kombinovanom hranom - peletirana i Tubifex, druga grupa animalnom hranom – Tubifex sp. i treća grupa peletiranom hranom. Peletirana hrana, korišćena u ishrani riba, proizvedena je u fabrici Royal optima 1P, uvoz iz Italije. Animalna hrana - Tubifex sp.donošena je iz toka reke Lepenice. Na početku eksperimenta po akvarijumu nasađeno je 10 riba prosečne mase 5,52 g, odnosno 30 jedinki po formiranoj odgovarajućoj grupi ishrane. Ishrana riba zasnivala se na količini hrane koja iznosi 1-3 % ukupne mase riba u formiranoj grupi. Premda je masa riba u akvarijumima svih formiranih grupa bila ista, bila je predviđena ishrana istom količinom hrane. Prema literaturnim saznanjima, ali i iz godišnjih statističkih pregleda gotovo je sigurno da je proizvodnja linjaka u potpunosti potisnuta. Najčešće se ističe da je razlog za napuštanje gajenja slaba konverzija hrane. Poređenje dužinskih i težinskih parametara na početku i na kraju eksperimenta pokazuje značajno povećanje mase riba, kao i njihove totalne dužine. Na osnovu dobijenih rezultata konstatovano je da je linjak riba koja se može uspešno gajiti u laboratorijskim uslovima, kao i da su najbolji prirast ostvarile jedinke hranjene kombinovanom hranom - peletiranom i animalnom

Determination of Total and Individual Anthocyanins in Raspberries Grown in South Serbia

Raspberry fruits of four cultivars (Willamette, Meeker, Polana and Malling Promise) grown in South Serbia were analyzed for total and individual anthocyanins. High-performance liquid chromatography with diode array detection (HPLC-DAD) was used to study individual anthocyanins. The major anthocyanin was cyanidin-3-sophoroside, followed by cyanidin-3-glucosylrutinoside, cyanidin-3-glucoside, pelargonidin-3-sophoroside and cyanidin-3-rutinoside. The monomeric anthocyanin contents of the acidified 80% methanol extracts were determined using the pH-differential method. The highest total and individual anthocyanins contain raspberries cv. Willamette, followed by cvs. Meeker, Polana and Malling Promise

The importance of combined NGS and MLPA genetic tests for differential diagnosis of maturity onset diabetes of the young

Introduction: Maturity onset diabetes of the young (MODY) is a rare form of monogenic diabetes. Being clinically and genetically heterogeneous, it is often misdiagnosed as type 1 or type 2 diabetes, leading to inappropriate therapy. MODY is caused by a single gene mutation. Thirteen genes, defining 13 subtypes, have been identified to cause MODY. A correct diagnosis is important for the right therapy, prognosis, and genetic counselling.Material and methods: Twenty-nine unrelated paediatric patients clinically suspected of having MODY diabetes were analysed using TruSight One panel for next-generation sequencing (NGS) and multiplex ligation-dependent probe amplification (MLPA) assay.Results: In this study we identified variants in MODY genes in 22 out of 29 patients (75.9%). Using two genetic tests, NGS and MLPA, we detected both single nucleotide variants and large deletions in patients. Most of the patients harboured a variant in the GCK gene (11/22), followed by HNF1B (5/22). The rest of the variants were found in the NEUROD1 and HNF1A genes. We identified one novel variant in the GCK gene: c.596T>C, p.Val199Ala. The applied genetic tests excluded the suspected diagnosis of MODY in two patients and revealed variants in other genes possibly associated with the patient’s clinical phenotype.Conclusions: In our group of MODY patients most variants were found in the GCK gene, followed by variants in HNF1B, NEUROD1, and HNF1A genes. The combined NGS and MLPA-based genetic tests presented a comprehensive approach for analysing patients with suspected MODY diabetes and provided a successful differential diagnosis of MODY subtypes

High-normal blood pressure, functional capacity and left heart mechanics: is there any connection?

Objective. To evaluate the relationship between functional capacity and left ventricular (LV) and left atrial (LA) myocardial deformation, assessed by two- and three-dimensional (2DE and 3DE) strain analysis, in subjects with high-normal blood pressure (BP). Methods. This cross-sectional study included 64 subjects with optimal BP and 75 subjects with high-normal BP of similar gender and age. All the subjects underwent a complete 2DE and 3DE examination and cardiopulmonary exercise testing. Results. 3DE global longitudinal strain was significantly lower in the group with high-normal BP than in the optimal BP group (- 20.1 ± 2.4 vs - 18.5 ± 2.3%, p < 0.001). Similar results were obtained for 3DE global circumferential strain (- 21.8 ± 2.6 vs - 19.3 ± 2.4%, p < 0.001), as well as for 3DE global radial strain (45.1 ± 8.8 vs 42.3 ± 7.2%, p = 0.042), and 3DE global area strain (- 30.1 ± 4.2 vs - 28.1 ± 3.8%, p < 0.001). LV twist was similar between the observed groups, whereas untwisting rate was significantly decreased in the subjects with high-normal BP (- 123 ± 30 vs - 112 ± 26°/s, p = 0.023). Peak VO2 was significantly lower in the high-normal BP group (30.8 ± 4 vs 28.3 ± 3.7 ml/kg/min, p < 0.001). 2DE LV ejection fraction (β = 0.38, p = 0.014), 2DE global longitudinal strain (β = 0.35, p = 0.019) and 3DE global longitudinal strain (β = 0.27, p = 0.042) were independently associated with peak VO2. Conclusion. LV and LA mechanics, as well as functional capacity are significantly impaired in the subjects with high-normal BP. LV and LA myocardial deformations are associated with peak oxygen uptake

ODREĐIVANJE HEMIJSKOG SASTAVA RAZLIČITIH EKSTRAKATA ČUVARKUĆE

Sempervivum tectorum has a similar effect as aloe vera, which is known in the treatment of various skin diseases. This herb is considered one of the safest remedies for a wide range of skin diseases. Due to its anti-inflammatory and antiseptic properties, it also serves as an excellent first aid for burns, stings and bites, because it provides quick relief and calming. Freshly squeezed juice from the leaves of the houseplant is used in the treatment of nervous disorders, epilepsy and restless dreams. The leaves are edible and can be used as an addition to salads or stews. They are not particularly tasty, but as they are rich in water, they can be put in a juicer together with other fruits or vegetables and become a refreshing drink. It is used in folk herbal medicine and as a medicine. The aim of this study was to determine the moisture content, total extracted substances, extract density, vitamin C, organic acids and proteins in house extracts.Publishe