Prognostic significance of aberrant CpG islands methylation in human follicular and diffuse large B-cell lymphoma

Difuzni B-krupnoćelijski limfom (DBKL) i folikularni limfom (FL), predstavljaju najučestalije entitete nehočkinovih limfoma i zajedno čine oko 50 % svih B-ćelijskih neoplazija. To su klinički i biološki heterogena oboljenja, sa visoko varijabilnim odgovorom na terapiju i uprkos značajnom napretku u terapiji, u velikom broju slučajeva ostaju neizlečiva. Aktuelni terapijski pristupi su praćeni brojnim neželjenim efektima i dovode do dugotrajne remisije i preživljavanja kod svega 50 % bolesnika. Najznačajniji klinički prediktor ishoda bolesti do danas je internacionalni prognostički indeks (IPI za DBKL, odnosno, FLIPI za FL), koji obuhvata odgovarajuće kliničke prognostičke parametre. Uprkos tome, oboleli koji pripadaju istim IPI/FLIPI kategorijama ispoljavaju značajne razlike u odgovoru na terapiju i preživljavanju, što ukazuje na postojanje značajne biološke heterogenosti u okviru svake IPI/FLIPI kategorije. Zbog toga je neophodno pronalaženje novih prognostičkih molekularnih parametara, koji bi omogućili precizniju podelu obolelih u grupe različitog stepena rizika i izbor odgovarajuće terapije. Primenom PCR metode specifične za metilaciju (MSP), ispitivan je metilacioni status p16, p15, MGMT i DAPK gena kod obolelih od DBKL i FL, kao bi se utvrdio njihov potencijalni prognostički značaj. Kod obolelih od DBKL, metilacija nijednog gena nije pokazala prognostički značaj, mada je uočena tendencija ka akumulaciji metilacije p15 gena kod ispitanika sa povoljnijim kliničko-patološkim karakteristikama. Kod obolelih od FL metilacija p16, p15 i MGMT gena bi mogla da ima određeni prognostički značaj, ukoliko se kombinuje sa FLIPI, gradusom tumora, odnosno starosnim dobom obolelihi. Istovremena metilacija MGMT i DAPK gena bi mogla da ukaže na grupu obolelih potencijalno hemorezistentnih na primenjenu hemoterapiju i sklonu recidivima bolesti.Diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) are the most comon subgroups of non-Hodgkin’s lymphoma and comprise approximately 50 % of all cases. They represents clinically and biologically heterogeneous diseases, characterized with highly variable response to the treatment, and remain incurable despite the significant advances in therapy. Actual therapy regimens are agressive, and long-term remission and survival are ashieved in only 50 % of patients. The strongest clinical predictor of outcome to date in DLBCL and FL patients is the International Prognostic Index (IPI for DLBCL and FLIPI for FL) which includes several clinico-pathological prognostic parameters. However, patients with identical IPI/FLIPI still exhibit marked variability in survival, suggesting the presence of significant biological heterogenity within the same risk category. So, it is important to investigate additional molecular markers in order to further stratify patients into diferent risk groups, choose appropriate treatement strategy and improve prognosis. In this study, methylation-specific polymerase chain reaction (MSP) was used to investigate the role and potential prognostic significance of the methylation status of p16, p15, MGMT and DAPK genes in patients with DLBCL and FL. No one of four examined genes showed prognostic significance in patients with DLBCL, though we observed a tendency toward accumulation of p15 methylation with favorable clinico-pathological parameters. However, in patients with FL, our results suggest that promoter methylation of p16, p15 and MGMT genes could have some prognostic value when used in combination with the FLIPI, tumor grade and patients age, respectively. Concurrent methylation of MGMT and DAPK genes could be the marker of tumor chemoresistance and disease recurrence

Prognostic significance of aberrant CpG islands methylation in human follicular and diffuse large B-cell lymphoma

Difuzni B-krupnoćelijski limfom (DBKL) i folikularni limfom (FL), predstavljaju najučestalije entitete nehočkinovih limfoma i zajedno čine oko 50 % svih B-ćelijskih neoplazija. To su klinički i biološki heterogena oboljenja, sa visoko varijabilnim odgovorom na terapiju i uprkos značajnom napretku u terapiji, u velikom broju slučajeva ostaju neizlečiva. Aktuelni terapijski pristupi su praćeni brojnim neželjenim efektima i dovode do dugotrajne remisije i preživljavanja kod svega 50 % bolesnika. Najznačajniji klinički prediktor ishoda bolesti do danas je internacionalni prognostički indeks (IPI za DBKL, odnosno, FLIPI za FL), koji obuhvata odgovarajuće kliničke prognostičke parametre. Uprkos tome, oboleli koji pripadaju istim IPI/FLIPI kategorijama ispoljavaju značajne razlike u odgovoru na terapiju i preživljavanju, što ukazuje na postojanje značajne biološke heterogenosti u okviru svake IPI/FLIPI kategorije. Zbog toga je neophodno pronalaženje novih prognostičkih molekularnih parametara, koji bi omogućili precizniju podelu obolelih u grupe različitog stepena rizika i izbor odgovarajuće terapije. Primenom PCR metode specifične za metilaciju (MSP), ispitivan je metilacioni status p16, p15, MGMT i DAPK gena kod obolelih od DBKL i FL, kao bi se utvrdio njihov potencijalni prognostički značaj. Kod obolelih od DBKL, metilacija nijednog gena nije pokazala prognostički značaj, mada je uočena tendencija ka akumulaciji metilacije p15 gena kod ispitanika sa povoljnijim kliničko-patološkim karakteristikama. Kod obolelih od FL metilacija p16, p15 i MGMT gena bi mogla da ima određeni prognostički značaj, ukoliko se kombinuje sa FLIPI, gradusom tumora, odnosno starosnim dobom obolelihi. Istovremena metilacija MGMT i DAPK gena bi mogla da ukaže na grupu obolelih potencijalno hemorezistentnih na primenjenu hemoterapiju i sklonu recidivima bolesti.Diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) are the most comon subgroups of non-Hodgkin’s lymphoma and comprise approximately 50 % of all cases. They represents clinically and biologically heterogeneous diseases, characterized with highly variable response to the treatment, and remain incurable despite the significant advances in therapy. Actual therapy regimens are agressive, and long-term remission and survival are ashieved in only 50 % of patients. The strongest clinical predictor of outcome to date in DLBCL and FL patients is the International Prognostic Index (IPI for DLBCL and FLIPI for FL) which includes several clinico-pathological prognostic parameters. However, patients with identical IPI/FLIPI still exhibit marked variability in survival, suggesting the presence of significant biological heterogenity within the same risk category. So, it is important to investigate additional molecular markers in order to further stratify patients into diferent risk groups, choose appropriate treatement strategy and improve prognosis. In this study, methylation-specific polymerase chain reaction (MSP) was used to investigate the role and potential prognostic significance of the methylation status of p16, p15, MGMT and DAPK genes in patients with DLBCL and FL. No one of four examined genes showed prognostic significance in patients with DLBCL, though we observed a tendency toward accumulation of p15 methylation with favorable clinico-pathological parameters. However, in patients with FL, our results suggest that promoter methylation of p16, p15 and MGMT genes could have some prognostic value when used in combination with the FLIPI, tumor grade and patients age, respectively. Concurrent methylation of MGMT and DAPK genes could be the marker of tumor chemoresistance and disease recurrence

Role of transforming growth factor-β1 in breast carcinogenesis

The main objective of this presentation is to review current knowledge regarding molecular mechanisms of Transforming Growth Factor-β1(TGF-ß1) action in breast carcinogenesis. In addition, our recent results will be presented on TGF-ß1 gene polymorphism and its relationship to TGF-ß1 secretion in breast cancer (BC) patients. Special focus will be made on potential clinical applicability of TGF-ß1 as a putative diagnostic, prognostic or predictive tool in BC detection and treatment. TGF-ß1 has a complex multifunctional profile, with tumor suppressive effects in early stages of breast carcinogenesis, but progressive dominance of tumor promoting effects with transition to more advanced malignant states. Clarification of molecular mechanisms that control parallel processing of these opposing TGF-ß1 activities might suggest new approaches for shifting the balance in favor of net tumor suppression. Now, a major challenge remains in more precisely defining TGF-ß1 signaling pathways and their cancer-related alterations. Current dogma views human tumorigenesis as a molecular disruption of normal physiology through genetic, epigenetic, or somatic alterations. The genetic model offers biological plausibility to epidemiological studies that link the TGF-ß1 gene polymorphism, at codon 10 due to Leu10Pro substitution in the signal peptide, with the risk of developing BC. The somatic mutations approach, provides an explanation for the TGF-ß1 overexpression in advanced BC through mutations acquired in the components of Smad-mediated TGF-ß1 signaling pathway. The available results indicate decreased TßRII (TGF-ß1 receptor-type II) expression, rare TßRII gene mutations, but no mutations in Smad2 and Smad4 genes, in advanced BC patients.

Multiple Roles of the RUNX Gene Family in Hepatocellular Carcinoma and Their Potential Clinical Implications

: Hepatocellular carcinoma (HCC) is one of the most frequent cancers in humans, characterised by a high resistance to conventional chemotherapy, late diagnosis, and a high mortality rate. It is necessary to elucidate the molecular mechanisms involved in hepatocarcinogenesis to improve diagnosis and treatment outcomes. The Runt-related (RUNX) family of transcription factors (RUNX1, RUNX2, and RUNX3) participates in cardinal biological processes and plays paramount roles in the pathogenesis of numerous human malignancies. Their role is often controversial as they can act as oncogenes or tumour suppressors and depends on cellular context. Evidence shows that deregulated RUNX genes may be involved in hepatocarcinogenesis from the earliest to the latest stages. In this review, we summarise the topical evidence on the roles of RUNX gene family members in HCC. We discuss their possible application as non-invasive molecular markers for early diagnosis, prognosis, and development of novel treatment strategies in HCC patients

Мedicinske misije u Balkanskim ratovima kroz prizmu istorijskih izvora

In the first half of the 19th century, the Principality of Serbia began to work on developing the health service and educating medical staff by sending students abroad to study medicine, because there was no Medical Faculty. The aim of the paper is to present the participation of foreign medical missions in providing assistance to Serbia during the Balkan wars. Method of historical analysis by using the primary and the secondary data sources was applied. The new wave of war, which followed in the second decade of the 20th century, pointed to the lack of personnel, medical and pharmacy equipment. The lack of medical staff was compensated by medical missions from abroad. During the First Balkan War, Serbia was visited by 16 medical missions from all parts of Europe and Scandinavia with a total of 185 doctors. Only Russia sent eleven medical missions and thus took the leading position in providing assistance. The mission consisted of 28 doctors, 11 intendants, 2 pharmacists, 86 mercy nurses and 160 nurse assistants. In the Second Balkan War, England and Scotland, Denmark, Belgium, the Netherlands, Sweden, Norway, Germany, Russia, Hungary and Austria provided medical assistance with 85 doctors (1). All missions were consistent with love and dedication in saving lives, financing medical staff, medical supplies and hospital equipment. The courage and humanity of foreign missions was awarded the Order of Saint Sava.Kneževina Srbija je u prvoj polovini 19. veka počela da radi na razvijanju zdravstvene službe i edukaciji lekarskog kadra slanjem studenata u inostranstvo na studije medicine, jer Medicinskog fakulteta nije bilo u Kneževini. Cilj rada je da se prikaže učešće stranih medicinskih misija u pružanju pomoći Srbiji za vreme balkanskih ratova.U radu je primenjena metoda istorijske analize uz korićenje primarnih i sekundarnih izvora podataka. Novi ratni talas, koji je usledio u drugoj deceniji 20. veka, ukazao je na nedostatak personala, sanitetske i apotekarske opreme. Nedostatak medicinskog personala bio je nadoknađen medicinskim misijama iz inostranstva. Za vreme Prvog balkanskog rata zabeležen je dolazak 16 medicinskih misija iz Evrope i Skandinavije, sa ukupno 185 lekara. Rusija je uputila jedanaest sanitetskih misija i time zauzela vodeće mesto u pružanju pomoći. Misiju je činilo 28 lekara, 11 ekonoma, 2 farmaceute, 86 milosrdnih sestara i 160 bolničara. U Drugom balkanskom ratu Engleska i Škotska, Danska, Belgija, Holandija, Švedska, Norveška, Nemačka, Rusija, Mađarska i Austrija su pružile medicinsku pomoć sa 85 lekara (1). Sve misije su sa ljubavlju i predanošću bile dosledne u spašavanju života, finansiranju medicinskog kadra, sanitetskog materijala i bolničke opreme. Hrabrost i humanist stranih misija odlikovana je dodeljivanjem Ordena Svetog Save.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Variability of the HCV core region and host genetic and epigenetic factors can predict the response to pegylated interferon/ribavirin therapy in genotype 1b hepatitis C patients from Serbia

Variations in the hepatitis C virus (HCV) core sequence have been related to disease progression and response to antiviral therapy. Previously we showed that the methylation status of RASSF1A and p16 genes, and IL28B genotypes affects the response to pegylated interferon/ribavirin (PEG-IFN/RBV) therapy. Herein we investigated whether amino acid (aa) substitutions in the HCV core region alone or in combination with IL28B genotypes and RASSF1A/p16 methylation affect the response to PEG-IFN/RBV therapy and liver disease progression. Among 29 examined patients, we found no association between single aa substitutions and response to therapy. However, we observed that patients with the HCV core aa substitution at position 75 and CT/TT IL28B genotypes were non-responders (NR), (P=0.023). Moreover, these patients had unmethylated RASSF1A. In contrast, most patients (75%) with aa substitutions at position 91 and CC IL28B genotype achieved sustained virologic response (SVR), (P=0.030), and 70% of them had methylated RASSF1A gene. Our results suggest that combined analysis of aa substitutions in the core protein, the IL28B rs12979860 polymorphism, and the methylation status of the RASSF1A gene may help in predicting treatment response to PEG-IFN/RBV in genotype 1b chronic hepatitis C patients

Estrogen Receptor Beta: The Promising Biomarker and Potential Target in Metastases

The discovery of the Estrogen Receptor Beta (ERβ) in 1996 opened new perspectives in the diagnostics and therapy of different types of cancer. Here, we present a review of the present research knowledge about its role in endocrine-related cancers: breast, prostate, and thyroid, and colorectal cancers. We also discuss the reasons for the controversy of its role in carcinogenesis and why it is still not in use as a biomarker in clinical practice. Given that the diagnostics and therapy would benefit from the introduction of new biomarkers, we suggest ways to overcome the contradictions in elucidating the role of ERβ.Special Issue "Biomarkers in Hematological and Oncological Malignancies: Identification, Validation and Involvement in Molecular Pathways

Methylation-specific PCR: four steps in primer design

Methylation-specific PCR (MSP) is still the method of choice for a single gene methylation study. The proper design of the primer pairs is a prerequisite for obtaining reliable PCR results. Despite numerous protocols describing the rules for MSP primer design, none of them provide a comprehensive approach to the problem. Our aim was to depict a workflow for the primer design that is concise and easy to follow. In order to achieve this goal, adequate tools for promoter sequence retrieval, MSP primer design and subsequent in silico analysis are presented and discussed. Furthermore, a few instructive examples regarding a good versus a poor primer design are provided. Finally, primer design is demonstrated according to the proposed workflow. This article aims to provide researchers, interested in a single gene methylation studies, with useful information regarding successful primer design

RASSF1A and p16 promoter methylation and treatment response in chronic hepatitis C genotype 1b patients treated with pegylated interferon/ribavirin

Prevention of chronic hepatitis C (CHC) and its complications is based on antiviral therapy and early detection of reliable molecular markers in persons under risk. We investigated whether the methylation status of RASSF1A and p16 genes, alone or in combination with host and viral factors, affects the response to therapy with pegylated interferon/ribavirin (PEG-IFN/RBV). Methylation-specific polymerase chain reaction (MSP) was used to determine the methylation status of the target promoter sequences of RASSF1A and p16 in circulating-free DNA from the peripheral blood of 49 patients with CHC genotype 1b. The methylation status of the examined genes did not affect the response to therapy. However, the simultaneous presence of either RASSF1A or p16 methylation and the CC genotype of IL28B was significantly related to a sustained virologic response (P=0.009 and P=0.032, respectively). After Bonferroni correction, only the result concerning the RASSF1A gene remained significant (P<0.0125). Methylation of RASSF1A was associated with the CC genotype of the IL28B gene (P=0.024) and a higher viral load (≥400 000 IU/mL, P=0.009). Our results suggest that combined analysis of RASSF1A gene methylation and IL28B rs12979860 polymorphism could potentially help in the prediction of therapy response in CHC genotype 1b patients

Ti-SLActive and TiZr-SLActive Dental Implant Surfaces Promote Fast Osteoblast Differentiation

A primary goal in modern surface modification technology of dental implants is to achieve biocompatible surfaces with rapid but controlled healing which also allow health and longevity of implants. In order to realize all, understanding of osseointegration phenomena is crucial. Although Ti-SLA, Ti-SLActive and TiZr-SLActive surfaces have been successfully used in clinical implantology and were shown to notably reduce the primary healing time, available in vitro studies are sparse and do not concern or explore the mechanism(s) involved in human osteoblast behavior on these surfaces. Ti-SLA, Ti-SLActive, TiZr-SLActive, Ti cp, Ticer and Cercon surfaces were used. Osteoblast proliferation, cell cluster formation, morphological changes, induction of autophagy, nitric oxide (NO), reactive oxygen species/reactive nitrogen species (ROS/RNS) formation, osteocalcin (OC), bone sialoprotein (BSP) and collagen type I (Col-1) affected by various surfaces were analyzed. These surfaces induced formation of mature osteoblasts caused by elevated oxidative stress (ROS) followed by overexpression of osteoblast maturation key molecule (NO), with different intensity however. These mature osteoblasts induced upregulation of OC, BSP and Col-1, activating PI3/Akt signalling pathway resulting in autophagy, known as an important process in differentiation of osteoblast cells. Additional distinctive subpopulation identified on Ticer, Ti-SLA (after 5 days), Ti-SLActive and TiZr-SLActive surfaces (after 2 days) were forming cell clusters, essential for bone noduli formation and mineralisation. The results suggest that Ti- and TiZr-SLActive possess advanced properties in comparison with Ticer and Ti-SLA manifested as accelerated osteoblast differentiation. These effects could explain already known fast osseointegration of these surfaces in vivo