Uloga citomegalovirusa i Epštajn-Bar virusa u nastanku i progresiji karcinoma pljuvačnih žletda

Introduction: Salivary gland carcinomas (SGC) are rare tumors characterized by an enormous morphological diversity between different subtypes going along with diverse clinical courses. The etiology of SGC is still unknown, although a correlation has been shown between the occurrence of SGC and some environmental factors, however, only a very small percentage of malignancies develop as a direct result of these factors. There is growing evidence that infectious agents are frequently associated with human cancer. Recent studies show that Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) CMV infections are associated with various epithelial malignancies. It has been shown that in various tissues CMV and EBV could increase the expression of IL-6, NFκB, MMP-2 and MMP-9. These proteins are associated with certain types of cancer. We hypothesized that CMV and EBV could cause inflammation and deregulation of genes involved in cell proliferation and could induce the development of salivary gland cancer. Aims: The aims of this study were to determine the prevalence of CMV and EBV in SGC tissues, to compare the immunohistochemical and nested PCR detection of CMV and EBV in SGC, to investigate the viral protein expression in different histological types of salivary gland cancer, to investigate the immunohistochemical expression of prognostic markers IL6, MMP2, MMP9 and NFκB, to investigate the association of the presence of CMV and EBV and the immunohistochemical expression of these prognostic markers and to analyze the polymorphisms of IL6, MMP2 and MMP9 genes and investigate the association of the polymorphisms and the corresponding protein expression. Material and Methods: This cross-section study included 93 patients diagnosed with salivary gland cancer (SGC), surgically treated at the Clinic of Otorhinolaryngology and Maxillofacial Surgery, Clinical Center of Serbia from 2004 to 2013. For the immunohistochemical analysis, the control consisted of healthy salivary gland tissue from 20 autopsy cases with no malignancies and salivary gland pathology...Uvod: Karcinomi pljuvačnih žlezda su retki tumori koji u različitim histološkim tipovima pokazuju spektar različitih morfoloških promena koje odgovaraju različitom kliničkom ispoljavanju i toku. Etiologija karcinoma pljuvačnih žlezda je još uvek nepoznata. Brojna istraživanja su pokazala značajan uticaj faktora sredine na njihov nastanak i progresiju, ali je samo za mali procenat jasno pokazano da je tumor nastao upravo direktnim uticajem nekog od faktora sredine. Sve je više istraživanja koja ukazuju na čestu i tesnu povezanost infektivnih agenasa sa pojavim maligniteta. Novija istraživanja pokazala su direktnu povezanost infekcija Citomegalovirusom (CMV) i Epštajn-Barr virusom (EBV) sa nastankom različitih oblika karcinoma. U različitim tkivima pokazano je da CMV i EBV mogu uzrokovati povećanje ekspresije IL-6, NFκB, MMP-2 i MMP-9, proteina koji su direktno povezani sa određenim tipovima karcinoma. Naša hipoteza je bila da CMV i EBV mogu dovest do zapaljenja i do deregulacije gena ćelijske proliferacije, te uticati na nastanak i razvoj karcinoma pljucačnih žlezda. Ciljevi: Ciljevi ove studije bili su da se utvrdi prisustvo CMV i EBV u tkivu karcinoma pljuvačne žlezde, da se uporedi immunohistohemijska i nested PCR detekcija CMV i EBV u karcinomu pljuvačne žlezde, da se ispita ekspresija virusnih proteina u različitim histološkim tipovima karcinoma pljuvačnih žlezda, da se ispita imunohistohemijska ekspresija prognostičkih markera IL6, MMP2, MMP9 i NFκB, da se ispita povezanost prisustva CMV i EBV sa imunohistohemijskom ekspresijom pomenutih prognostičkih markera i da se uradi analiza polimorfizma IL6, MMP2 i MMP9 gena i ispita asocijacija polimorfizama i odgovarajuće ekspresije proteina. Materijal i metode: Ova studija preseka obuhvatila je 93 pacijenta kod kojih je dijagnostikovan karcinom pljuvačne žlezde, koji su operisani na Klinici za otorinolaringologiju i maksilofacialnu hirurgiju, Kliničkog centra Serbije u periodu od 2004. do 2013. Detekcija CMV i EBV analizirana je imunohistohemijskom i nested PCR metodom..

Detection and sampling methods for isolation of Candida spp. from oral cavities in diabetics and non-diabetics

The purpose of this study was to detect Candida spp. on the tongue and in the subgingival sites in healthy and type 2 diabetes (T2D) patients with chronic periodontitis (CP), and to compare the accuracy of sampling methods. This study included 131 patients divided into four groups: healthy control (group A), nondiabetics + CP (Group B), diabetics with good metabolic control + CP (group C) and diabetics with poor glycoregulation + CP (Group D). Cotton swab samples from tongue and subgingival samples were obtained from each patient with help of sterile paper points and a sterile curette. Swab cultures were made on Sabouraud dextrose agar. The number of CFUs was counted. The sampling methods for subgingival plaque were compared by Receiving Operator Curve (ROC). The presence of Candida spp. on the tongue was statistically significant among groups (group D vs. others three groups:.2: p lt 0.005 for each group). Positive findings of subgingival Candida spp. did not differ among the groups. There were no significant differences in the quantification of Candida spp., neither on the tongue, nor in the subgingival samples. 17.2% of diabetic patients revealed the presence of Candida spp. in the subgingival samples, with negative finding on tongue. There was a significant difference in the sampling methods for subgingival plaque (p = 0.000). Candida spp. is more prevalent on the tongue of diabetics. The sampling of subgingival plaque by a sterile curette is more accurate than with paper points. Subgingival plaque may represent a reservoir of commensals. It is necessary to standardize the sampling of subgingival plaque

Rresence of different Candida species at denture wearers with type 2 diabetes and clinically healthy oral mucosa: Pilot study

Background/Aim: The aim of this study was to examine prevalence of different Candida spp. at diabetics and nondiabetics wearing dentures without clinical signs of Denture Stomatitis (DS) and to study if some local and systematic factors are confounders for harboring Candida at these subjects. Material and Methods: Total of 60 subjects wearing partial or complete upper acrylic denture having at least half of palatal mucosa covered by denture were selected and stratified into three experimental groups: systematically health subjects; patients with diagnosed Type 2 Diabetes (T2D) and good glycoregulation; and T2D subjects with poorly regulated blood sugar level. Cotton swab samples were obtained from each patient from hard palate mucosa and denture surface. Swab cultures were made on Sabouraud dextrose agar and ChromAgar Media for distinciton of various Candida spp. Density growth was also measured. Results: Frequency of Candida spp. findings were similar between groups. At healthy subjects, only C.albicans was detected. At diabetics, C.albicans was the most common isolated species, followed by C.glabrata and C.tropicalis. Negative finding of yeasts on palatal mucosa, but positive on denture surface were detected at all groups, with the highest frequency (33.4%) at diabetics with poor glycoregulation. Denture surface was heavier colonized than hard palate mucosa. Duration of diabetes in years were only independent predictors for harboring Candida spp. at denture surface (Exp B=1.186, CI=1.047-1.344, p=0.007). Conclusions: Prosthesis of denture wearers without DS may serve as reservoir of Candida spp. Presence of more pathogenic and resistant non-albicans species are related to diabetics, even without clinical signs of DS

Geographic tongue: Does Candida play a role in its pathogenesis

Background/Aim: Geographic tongue (GT), a benign self-limiting condition (inflammation) is commonly seen in practice. Although quite easy for clinical diagnosis, in clinical practice it is commonly misdiagnosed and treated as Candida infection. The main aim of this study was to compare the prevalence of Candida spp. on tongue in patients with GT and subjects with clinically healthy oral mucosa. Additional aims were to evaluate subjective symptoms, uncontrolled usage of prescribed or non-prescribed antifungal topical medications and the presence of the cancerophobia or other health concerns in patients with GT. Material and Methods: A total of 70 subjects were divided into two groups: Group B - patients with diagnosed Geographic tongue and group A - aged and gender matched controls with clinically health oral mucosa. Anamnestic charts designed for this study included information about symptoms (measured by Visual Analogue Scale), previous knowledge or fear about presence of GT and received therapy for this condition. Detection of Candida spp. from tongue was done using sterile cotton swab and Sabouraud dextrose agar. Results: At the time of this study, 18 (52.94%) of subjects with GT were aware of having this condition and even 12 of them (66.7%) used topical antifungal drugs prescribed by physician. Also, 66.7% of them experienced fear about the presence of GT at some moment. Detection of Candida spp. was similarly distributed in both groups (22.2% in group A and 17.6% in group B). Conclusions: Geographic tongue has frequently been treated by topical antifungal drugs. In this study, GT was not associated with presence of Candida spp

Cymbopogon citratus essential oil: an active principle of nanoemulsion against Enterococcus faecalis root canal biofilm

Aim: The objective was to formulate and characterize the nanoemulsion based on Cymbopogon citratus oil, intended for use in infected teeth root canal therapy. The investigation of the antioxidant and antibiofilm potential toward Enterococcus faecalis was aimed as well. Materials & methods: Characterization of oil (by GC/MS analysis) and nanoemulsion (by dynamic light scattering instrument), and determination of antibacterial (by microdilution assay), antibiofilm (by crystal violet assay) and antioxidant properties (by 2,2-diphenyl-1-picryl-hydrazyl-hydrate and thiobarbituric acid assay methods) were provided. Antibiofilm efficacy of irrigation procedure including nanoemulsion was screened on extracted teeth (by CFU-counting assay). Results: Notable antibacterial and antibiofilm activity, both against forming and preformed biofilms of oil, was observed. Irrigation involved nanoemulsion showed remarkable antibiofilm potential. Both substances induced some antioxidant activity. Conclusion: Results encourage further research with the aim of application of the nanoemulsion in dental practice

Antimicrobial and Osteogenic Effects of Collagen Membrane Decorated with Chitosan–Nano-Hydroxyapatite

Collagen membranes are routinely used in oral surgery for bone regeneration. Despite their numerous advantages, such as stimulating bone growth, bacterial contamination still remains one of the disadvantages of membrane use. Thus, we assessed the biocompatibility and osteogenic and antibacterial properties of a collagen membrane (OsteoBiol) modified with chitosan (CHI) and hydroxyapatite nanoparticles (HApNPs). Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR FT-IR), X-ray powder diffraction (XRD), and field emission scanning electron microscopy (FE-SEM) were performed for membrane characterization. Biocompatibility was assessed on dental pulp stem cells (DPSCs) by an MTT assay, while the osteogenic effect was assessed by an ALP activity assay and qPCR analysis of osteogenic markers (BMP4, ALP, RUNX2, and OCN). Antimicrobial properties were investigated by counting colony-forming units (CFUs) of Streptococcus mitis, Porphyromonas gingivalis, and Fusobaterium nucleatum on membranes and in the surrounding medium. Membranes showed no cytotoxicity. ALP activity was higher and ALP, BMP4, and OCN genes were up-regulated in DPSCs on modified membranes compared to unmodified membranes. The CFUs were reduced on modified membranes and in the medium. Modified membranes showed great biocompatibility and a high osteoinductive effect. Additionally, they showed antimicrobial and antibiofilm effects against periopathogens. It can be concluded that the incorporation of CHI and hydroxyapatite nanoparticles in collagen membranes may be advantageous to promote osteogenesis and reduce bacterial adhesion

Biocompatible Germanium-Doped Hydroxyapatite Nanoparticles for Promoting Osteogenic Differentiation and Antimicrobial Activity

Hydroxyapatite (HAp) has been the main protagonist in the quest for an ideal biomaterial for regenerative medicine over the last half a century. To control its properties, this material has commonly been doped with chemical elements other than its natural stoichiometric constituents: Ca, O, P, and H. Here, we report on the first analysis of the biological response to germanium-doped hydroxyapatite (Ge-HAp). Cytotoxicity, osteogenic differentiation induction, and colony formation potential were measured on dental pulp stem cells, while the antimicrobial effect was assessed against Gram-negative Escherichia coli, Gram-positive methicillin-resistant Staphylococcus aureus (MRSA), and Candida albicans. All analyses were run in comparison to Ge-free HAp. Cell viability was inversely dependent on the nanoparticle concentration and incubation time. Adding Ge to HAp reduced cell viability relative to HAp after 24–72 h incubation periods, but the effect was reversed after longer incubations, when the viability of cells treated with low doses of Ge-HAp exceeded that of HAp-treated cells and became comparable with control culture. Both HAp and Ge-HAp induced mineral formation in the cell culture, but the effect was more pronounced for Ge-HAp. Likewise, relative to both control cells and cells exposed to HAp, Ge-HAp upregulated the expression of all three osteogenic markers analyzed, namely, alkaline phosphatase, RUNX2, and osteocalcin, exerting the key influence on osteogenesis in its early, differentiation stage. The colony formation capacity of stem cells, however, was impaired by HAp and even more so by Ge-HAp. The antimicrobial effect was dependent on the microorganisms tested. Thus, whereas the antimicrobial activity was absent against E. coli, it was evident against MRSA and C. albicans. While the antibacterial activity against MRSA was weakened by the addition of Ge to HAp, the antimycotic activity against C. albicans was intensified with the addition of Ge. These findings demonstrate a significant potential of Ge-doped HAp nanoparticles in regenerative medicine due to their pronounced biocompatibility, osteoinductivity, and antimicrobial activity

Microbiological analysis of primary infected root canals with symptomatic and asymptomatic apical periodontitis of young permanent teeth

Background/Aim: Understanding the composition of bacteria in infected root canals is important for ameliorating the treatment strategies that lead to the elimination of pathogens and infection control, but also prevent reinfection. Aim of this study was to investigate microbial composition of primary infected root canals with apical periodontitis of young permanent teeth, originating form school children in Serbia, and its association with clinical symptoms. Material and Methods: To determine the bacterial composition of infected root canals in children, 35 endodontic samples were obtained. The identification of cultured bacteria was performed by MALDI-TOF MS analysis. The presence or absence of clinical symptoms were recorded. Results: Facultative anaerobes were 2,2 times more frequent than obligate anaerobes. The most common facultative anaerobes belonged to following genera, Streptococcus (58 isolates), Actinomyces (10) and Enterococcus (8), while predominant obligate anaerobes, belonged to genera Veillonella (15), Prevotella (9) and Fusobacterium (8). The most common clinical isolates recovered from infected root canals with symptomatic apical periodontitis were Veillonella parvula (10) and Fusobacterium nucleatum (7), while from the asymptomatic ones, they were Streptococcus mitis/Streptococcus oralis (5). Prevalence of Parvimonas micra, Prevotella buccae and Streptococcus constellatus within the root canals might be associated to clinical symptoms. Conclusions: Species of genera Streptococcus and Veillonella were the most common isolates from primary infected root canals with apical periodontitis in Serbian school children. Facultative anaerobes were predominant over obligate anaerobes. The prevalence of obligate anaerobes was much higher in symptomatic compared to asymptomatic root canal infections. No specific bacterial strain might be associated to a single examined clinical symptom (pain, tenderness to percussion or swelling), but majority of the strains are associated to all of the examined three symptoms

The presence of periopathogenic bacteria in subgingival and atherosclerotic plaques- An age related comparative analysis

Introduction: There is a known connection between periodontitis and atherosclerosis and the presence of periopathogens in blood vessels. However, changes of the oral microflora related to the aging process and its possible effects on atherosclerosis, have yet to be analyzed. The aim of this study was to assess temporal changes in the frequency of periodontal bacteria in the subgingival plaque and in atherosclerotic blood vessels of patients with atherosclerosis. Methodology: The study included 100 patients with atherosclerosis and periodontitis, divided into two groups, below and over 60 years of age. Clinical examinations were performedand subgingival plaque specimens were collected as well as biopsy specimens from the following arteries: coronary (34), carotid (29), abdominal (10), femoral (10), mammary (13) and iliac (4). Subgingival and artery specimens were subjected to PCR detection of 5 major periodontal pathogens: Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Aggregatibacter actinomycetemcomitans (Aa), Tannerella forsythensis (Tf) and Treponema denticola (Td). Results: Tf was the most and Td the least frequent bacteria in both age groups and in both types of samples. The frequencies of bacteria in subgingival versus atherosclerotic samples were: Tf (76%: 53%), Pi (71%: 31%), Pg (60%: 38%), Aa (39%: 14%) and Td (21%: 6%). Only Aa and Pi showed a significant difference of prevalence between younger and older patients. The most colonized artery was a. coronaria, followed by a. carotis, a. abdominalis, a. mammaria, and a. femoralis. Conclusions: Patient's age and the distance of a given blood vessel from the oral cavity influenced microbiological findings in the atherotic plaque

Xемијска карактеризација и антимикробна активност орашастог плода и зелене љуске Juglans nigra l.

Juglans nigra (Black walnut) is a source of health-promoting biologically active compounds used in traditional medicine. The investigation of bioactive compounds in black walnut could lead to its broader application, as well as application of its by-products. Therefore, this study aimed to characterize J. nigra nut and green husk based on chemical analysis of their petroleum ether and ethanol extracts obtained by ultrasonic and reflux extraction methods, respectively. Different extract fractions were tested for their antimicrobial activities using Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), Gram-positive bacteria (Enterococcus faecalis Staphylococcus aureus) and yeast (reference strain and clinical isolates of Candida albicans). Ethanol extracts analysis, performed by High Performance Liquid Chromatography (HPLC), singled out the ellagic acid as the most dominant compound in nut (55.0 ± 1.3 × 10-3 kg m-3 ) and green husk (114.1 ± 0.5× 10-3 kg m-3 ) extracts. Non-polar compounds were evaluated using Gas Chromatography (GC) analysis of petroleum ether extracts. Juglans nigra nut and green husk contained two saturated fatty acids, palmitic acid (C16:0) and stearic acid (C18:0), then, monounsaturated fatty acids, palmitoleic (C16:1n-7), oleic (C18:1n-9), and vaccenic acid (C18:1n-7), as well as polyunsaturated fatty acids, linoleic (C18:2n-6), γ-linolenic (C18:3n-6), and α-linolenic (C18:3n-3) acids. Ethanol extracts of both J. nigra nut and green husk showed antimicrobial activity against C. albicans which is the most common cause of yeast infections.Juglans nigra (црни орах) као извор биолошки активних једињења користе се у традиционалној медицини. Истраживање биоактивних једињења присутних у црном ораху довело би до шире примене његових производа. Зато ова студија има за циљ да окарактерише орашаст плод и зелену љуску Ј. nigra на основу хемијске анализе њихових петролетарских и етанолних екстраката, добијених ултразвучним и рефлукс методама екстракције. Антимикробна активност различитих фракција екстракта је тестирана коришћењем Грам-негативних бактерија (Escherichia coli, Pseudomonas aeruginosa), Грамозитивних бактерија (Enterococcus faecalis Staphylococcus aureus) и квасница (референтни сој и клинички изолати Candida albicans). Анализа течном хроматографијом високих перформанси, издвојила је елагинску киселину као најдоминантније једињење у етанолним екстрактима орашастог плода (55,0 ± 1,3 × 10-3 kg m-3) и зелене љуске (114,1 ± 0,5 × 10-3 g m-3). Неполарна једињења су процењена применом гасне хроматографске анализе петролетарских екстракта. Орашаст плод и зелена љуска садрже две засићене масне киселине, палмитинску (C16:0) и стеаринску киселину (C18:0), затим мононезасићене масне киселине, палмитолеинску (C16:1n-7), олеинску (C18:1n-9), и вакценску киселину (C18:1n-7), као и полинезасићене масне киселине, линолну (C18:2n-6), γ-линоленску (C18:3n-6) и α-линоленску (C18: 3n-3) киселину. Етанолни екстракти орашастог плода и зелене љуске показали су антимикробну активност нa сојевима C. albicans која је најчешћи узрочник гљивичних инфекција