Correlation between different genotypes of human cytomegalovirus and Epstein-Barr virus and peri-implant tissue status.

BACKGROUND: The purpose of this study was to estimate the prevalence of different genotypes of human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) in peri-implantitis and mucositis sites, and to evaluate the correlation between herpesvirus presence and clinical parameters. METHODS: A total of 80 dental implants (mean time of loading, 4.16 ± 1.8 years) were evaluated during the course of the study (30 peri-implantitis, 25 mucositis and 25 healthy peri-implant sites). The following clinical parameters were assessed: visible plaque index, bleeding on probing, suppuration and probing depth. A polymerase chain reaction (PCR) assay was used to identify the presence of different HCMV and EBV genotypes in peri-implant tissue plaque samples. RESULTS: HCMV-2 was detected in 53.3% and EBV-1 in 46.6% of the 30 peri-implantitis sites evaluated. By contrast, HCMV-2 was not detected in healthy periodontal sites and EBV-1 was detected in one healthy site. A statistically significant correlation was found between the presence of HCMV-2 and EBV-1 genotypes and clinical parameters of peri-implantitis. CONCLUSIONS: The results from the present study confirmed the high prevalence of HCMV-2 and EBV-1 in the peri-implant tissue plaque of peri-implantitis sites and suggests a possible active pathogenic role of the viruses in peri-implantitis.

Prevalence of human cytomegalovirus and Epstein-Barr virus in subgingival plaque at peri-implantitis, mucositis and healthy sites. A pilot study.

This study evaluated the prevalence of human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) in peri-implantitis and mucositis sites and the correlation between herpesvirus and clinical parameters. Fifty-six dental implants (mean time of loading, 4.27±1.6 years) were evaluated (20 peri-implantitis, 18 mucositis, 18 healthy peri-implant sites.) The clinical parameters assessed were: visible plaque index (PI), bleeding on probing (BOP), suppuration (SUP), probing depth (PD). A polymerase chain reaction assay identified HCMV and EBV in subgingival plaque samples. The percent of sites with plaque and BOP was significantly higher around mucositis and peri-implantitis compared with healthy implants (p<0.05). The mean PD around the implants was significantly higher in peri-implantitis, followed by mucositis and healthy implants (p<0.05). HCMV was detected in 13 (65%) and EBV in 9 (45%) of the 20 peri-implantitis sites. HCMV was found in 1 of the 18 (6%) healthy periodontal sites and EBV in 2 (11%). A statistically significant correlation was found between presence of HCMV and EBV subgingivally and clinical parameters of peri-implantitis and healthy sites. These results confirm the high prevalence of HCMV and EBV in subgingival plaque of peri-implantitis sites and suggest the viruses have a possible active pathogenic role in peri-implantitis.

Germline VHL gene mutations in three Serbian families with von Hippel-Lindau disease.

Von Hippel-Lindau (VHL) disease is an autosomal dominantly inherited cancer predisposition syndrome due to germline mutations in the VHL tumor suppressor gene which is associated with virtually complete penetrance. The VHL syndrome has a highly variable phenotypic expressivity including retinal and CNS haemangioblastomas, pheochromocytomas, renal clear cell carcinomas, and multifocal cysts. In order to establish VHL gene testing, we analyzed three families affected by VHL disease, using SSCP mutation screening and DNA sequencing. Among 18 family members with and without clinical manifestations, eight cases with germline VHL mutations were detected. In family A, a c.490G>T/ p.Gly93Cys substitution was found. In family B, with pheochromocytoma only phenotype, we detected a previously not described c.463G>A/p.Val84Met replacement. Within this family, a prenatal diagnosis was also performed. Affected members of the third family with a VHL type 1 disease carried a c.475T>C/p.Trp88Arg exchange. All these mutations were located in exon 1 of the VHL tumor suppressor gene. Alterations in this hydrophobic region of the core beta domain of the VHL protein are known to have a variety of phenotypic consequences. We observed also intrafamiliar variation in time of onset and severity of the disease.

Localization of recognition site between transforming growth factor-beta1 (TGF-beta1) and TGF beta receptor type II: possible implications in breast cancer.

Although overexpression of TGF-beta1 protein has been demonstrated in advanced breast cancer (BC) patients, as well as in other solid tumours, the molecular mechanism of this process remains obscure. This paper proposes that a genetic/epigenetic alteration might occur in the TGF-beta1 gene, within the region coding for the recognition site with TGFbeta receptor type II, leading to a disruption of the ligand-receptor interaction and triggering the TGF-beta1 cascade-related BC progression. To establish the operational framework for this hypothesis, in the present study, this recognition site was identified by the Informational Spectrum Method (ISM) to comprise two TGF-beta1 peptides (positions 47-66 aa and 83-112 aa) and one receptor peptide at positions 112-151 aa of the extracellular domain of the receptor (TbetaRIIM). The TbetaRIIM locus was further evaluated by ISM-derived deletion analysis of the TbetaRII sequences. To provide experimental support for the proposed model, a pilot study of plasma TGF-beta1 analysis was performed in advanced BC patients (n = 8). Two commercial ELISA assays, one with specific alphaTGF-beta1 MAb (MAb) and other with TbetaRIIM as the immobilized phase, revealed pronounced differences in the pattern of plasma TGF-beta1 elevation. In MAb-profile, the TGF-beta1 increase was detected in 7 of 8 patients, whereas analogous TbetaRIIM-profile revealed the elevation in 3 of 8 patients, taking a 50% of maximal elevation as the cut-off value. These findings are consistent with the proposed aberration of TGF-beta1 ligand within the TbetaRII recognition site. Summarizing, this model system is a good starting point for further genetic studies, particularly on genetic/epigenetic alterations of sequences involved in TGF-beta1 and TbetaRIIM interaction, with putative prognostic value for breast cancer.

Analysis of 5' non-coding region in hepatitis C virus by single-strand conformation polymorphism and low-stringency single specific primer PCR.

Single-strand conformation polymorphism (SSCP) and low-stringency single specific primer (LSSP)-PCR in hepatitis C virus (HCV) genotyping were examined for informativeness and reliability. The analysis of HCV isolates included seven type 1 isolates, two type 2 isolates, and two type 3 isolates. We also analyzed five isolates that presented as mixed infections determined by type-specific PCR. Among mixed isolates, one isolate was 1a/1b and four isolates were 1b/3a. SSCP and LSSP-PCR were applied to the analysis of 5' non-coding region of HCV (-289 to -5) that contains genotype-specific sequences. Direct cycle sequencing of this region determined sequence divergences that define genotype and sequence alterations within the same genotype. Optimized conditions for the SSCP analysis clearly distinguished between genotypes 1, 2 and 3. In addition, the SSCP analysis detected sequence variants within the same genotype. However, the SSCP analysis and DNA sequencing did not confirm the presence of mixed infections. LSSP analysis, not previously employed in HCV genotyping, enabled clear distinction between genotypes 1, 2 and 3, however, this method did not differentiate between sequence variants within a genotype. Importantly, the LSSP profile demonstrated distinction between mixed infection isolates.

Influence of bioactive glass on changes in alveolar process dimensions after exodontia.

BACKGROUND: Preservation of the alveolar process after tooth extraction is desirable because it facilitates placement of endosseous implants and minimizes adverse esthetic results associated with fixed partial dentures. The purpose of this study was to evaluate the clinical effectiveness of bioactive glass used as a graft material combined with calcium sulfate used in the form of a mechanical barrier in preserving alveolar ridges after tooth extraction. METHODS: Sixteen patients who required extraction of 2 anterior teeth or bicuspids participated in the study (split mouth design). After tooth extraction and elevation of a buccal full-thickness flap, experimental sockets were filled with bioactive glass, which in turn was covered with a layer of calcium sulfate. Control sites did not receive any graft or calcium sulfate. Titanium pins served as fixed reference points for measurements. No attempt was made to advance the flap to cover the socket areas on control or experimental sites (open socket approach). Reentry surgeries were performed at 6 months. RESULTS: Reentry surgeries showed that experimental sites presented with (1) significantly more internal socket bone fill (6.43 +/- 2.78 mm vs 4.00 +/- 2.33 mm on control sites), (2) less (although not statistically significantly less) resorption of alveolar bone height (0.38 +/- 3.18 mm vs 1.00 +/- 2. 25 mm on control sites), and (3) similar degree of horizontal resorption of the alveolar bony ridge as compared with controls (3. 48 +/- 2.68 mm vs 3.06 +/- 2.41 mm on control sites). CONCLUSIONS: This study suggests that treatment of extraction sockets with a combination of bioactive glass and calcium sulfate is of some benefit in preserving alveolar ridge dimensions after tooth extraction.

Hematological toxicity associated with tiazofurin-influence on erythropoiesis.

In this study hematological toxicity was analyzed after the single and repeated applications of tiazofurin (TZF). Cellularity of bone marrow, spleen and peripheral blood was examined, spanning the period of fifty days after the initial application. Analysis of hematological parameters was performed by slightly modified conventional techniques. The fraction of erythroid series was monitored during the experiment. Presented data describe kinetics of damage and recovery of hemopoietic tissue. Our results indicate that the effect of tiazofurin on cellularity of bone marrow and spleen and on erythropoiesis is reversible and dose dependent within tested dose range and therapeutic regimes. Twenty days after the application normal function of hemopoietic tissues was restored. This approach and results can be useful in defining the timing for sequencing and combination therapy with tiazofurin.

DNA profiling of B chromosomes from the yellow-necked mouse Apodemus flavicollis (Rodentia, Mammalia).

Using AP-PCR-based DNA profiling we examined some structural features of B chromosomes from yellow-necked mice Apodemus flavicollis. Mice harboring one, two, or three or lacking B chromosomes were examined. Chromosomal structure was scanned for variant bands by using a series of arbitrary primers and from these, informative bands were selected. The selection criteria used were the ability to differentiate between individuals of the species, to detect markers common for both A and B chromosomes, and, importantly, to differentiate between A- and B-chromosome sets. In addition to primers, profiling conditions were found to be critical for meeting the selection criteria. Primers and analysis conditions that demonstrated structural characteristics unique to the B-chromosome set are described. These characteristics included variant bands as qualitative parameters and altered electrophoretic band intensities as quantitative distinctions estimated by integration of densitometric profiles of electrophoretograms. B chromosome-specific molecular markers are easy to detect by AP-PCR-based DNA profiling in the presence of a full set of A chromosomes. Models for the origin of yellow-necked mouse B chromosomes are discussed in the context of presented data.

Distribution of HCV genotypes among risk groups in Serbia.

Blood samples from 190 patients that were anti-hepatitis C virus (HCV) positive were genotyped and 165 were found to contain HCV-RNA. Genotyping was performed by PCR based on type-specific primers (117 isolates) and LiPA test (48 isolates) and verifying by sequencing. In Serbia, the most frequent genotype was 1b (49.1%), followed by genotype 3 (21.2%) and genotype 1a (8.5%). The frequency of genotypes 2 and 4 was below 5% and mixed infections were encountered in 9.1% of cases. Distribution of genotypes was monitored in different risk groups: intravenous drug abusers, patients under blood transfusion, patients with previous history of surgery, patients undergoing hemodialysis and those with unknown risk factors. Genotype distribution is essentially the same in all the groups, except for the patients undergoing hemodialysis and those with previous history of surgery where significant difference exists compared with the group with unknown route of transmission (p < 0.001 and p < 0.05, respectively). There exists significant age-dependent genotype 3 distribution in Serbian population (p < 0.01).

Germline mutation analysis of BRCA1 and BRCA2 genes in Yugoslav breast/ovarian cancer families.

The frequency of germline BRCA1 and BRCA2 mutations was tested in Yugoslav breast and breast/ovarian cancer families using combined heteroduplex/single-strand conformation polymorphism analysis for the entire coding region of both genes. Three different recurrent BRCA1 mutations (one 185delAG, one 3447del4 and two 5382insC) were identified in 4 of 12 families (33%), whereas no definite disease-causing alterations of BRCA2 was detected. Genotype analysis revealed a possible common founder effect for each 185delAG and 5382insC. The relatively high frequency of germline BRCA1 mutations determined in this panel of families confirms the important role of BRCA1 in disease predisposition in the Yugoslav population, while the lack of population specific founder and/or unique mutations show the need of further analysis of samples from this yet unexamined region of Europe.

The use of autogenous periosteal grafts as barriers for the treatment of intrabony defects in humans.

The purpose of this study was to evaluate the clinical effectiveness of a connective tissue graft including periosteum used as a barrier for guided periodontal tissue regeneration in interproximal bony defects. An open flap debridement of a comparable interproximal bony defect in the same patient was used as a control. This study was performed at 2 different clinical centers. Six paired defects were treated at one center, and 16 paired defects at the other. Reentry surgeries were performed at 6 months. Preoperative comparisons of control and experimental sites with respect to clinical parameters and osseous measurements were similar. Post-surgical experimental sites produced more gain in clinical attachment (1.25 mm on buccal and 1.25 mm on lingual sites at center A and 1.26 mm on buccal and 1.18 mm on lingual sites at center B) and osseous defect fill (1.84 mm on buccal and 2.00 mm on lingual sites at center A and 1.66 mm on buccal and 2.04 mm on lingual sites at center B) when compared to control sites. The results of this trial indicate that clinical resolution of interproximal periodontal defects can be obtained with periosteal grafts used as barriers.

A dental implant: aluminium trioxide exhibited no effect on mouse reproductive and mutanogenic potential.

Several diseases as well as trauma can affect the composition and integrity of periodontal tissues leading eventually to the destruction of connective tissue matrix and cells, loss of attachment and resorption of alveolar bone, often followed by tooth loss. Replacement of the missing tooth could then be provided by endosseous dental implants healing in a form of osseo- or fibrosteal integration to the alveolar bone. Aluminium oxide ceramics, a form of endosseous implant, allows osseointegration type of healing and has demonstrated excellent biocompatibility. However, potential aluminium toxicity has been implicated in the pathogenesis of a number of clinical disorders and for this reason we examined the reproductive and mutagenic effect of aluminium trioxide ceramic implant in experimental mice. 720 female and 45 fertile male BALB-cAn NCR mice were included in the study. 3 experimental groups of fertile male mice (15 for each group) were treated with an intraperitoneal injection of aluminium trioxide (1 g/ kg of body weight, group I), with ethyl-methane-sulphonate as a positive control (200 mg/kg, group II) and with Tween-80 (10 mg/kg as a negative control, Group III). Each of the labeled male mice fertilized previously uncoupled female mice during 8 weeks (a pair per week) to facilitate appropriate pre- and post-meiotic conditions of spermatogenesis to occur. Female mice were sacrificed with cervical dislocation at day 13 after fertilization. Immediately upon sacrifice the uterus was removed and the number of alive and healthy, or alive but mutated and/or dead embryos was computed to determine the dominant lethal or mutagenic effect. Animals treated with aluminium trioxide demonstrated similar effects on the reproductive and mutagenic capacity as the negative control, whereas the animals treated as positive controls exhibited significantly reduced reproductive and mutagenic capacity. Collectively, we concluded that aluminium trioxide has a very low rate of embryonal mortality and mutagenicity in mice. This finding is in general agreement with the biocompatibility of aluminium trioxide as an implant material.

Preservation of alveolar bone in extraction sockets using bioabsorbable membranes.

The purpose of this study was to evaluate the clinical effectiveness of a bioabsorbable membrane made of glycolide and lactide polymers in preserving alveolar ridges following tooth extraction using a surgical technique based on the principles of guided bone regeneration. Sixteen patients requiring extractions of 2 anterior teeth or bicuspids participated in the study (split-mouth design). Following elevation of buccal and lingual full-thickness flaps and extraction of teeth, experimental sites were covered with bioabsorbable membranes; control sites did not receive any membrane. Titanium pins served as fixed reference points for measurements. Flaps were advanced in order to achieve primary closure of the surgical wound. No membrane became exposed in the course of healing. Reentry surgeries were performed at 6 months. Results showed that experimental sites presented with significantly less loss of alveolar bone height, more internal socket bone fill, and less horizontal resorption of the alveolar bone ridge. This study suggests that treatment of extraction sockets with membranes made of glycolide and lactide polymers is valuable in preserving alveolar bone in extraction sockets and preventing alveolar ridge defects.

[Standards of physical fitness in Yugoslav military personnel]. / Norme fizicke pripremljenosti u Vojsci Jugoslavije.

With the aim to develop the standards of aerobic capacity as one of the basic elements of physical fitness of the military personnel, the authors worked out an original method, based on determination of energy requirements for different military activities, and on WHO principle for estimation of working capacity. According to the data on application of these standards in the group of 176 officers in the Yugoslav Armed forces, shown in the paper, even 53.4% og the officers have the aerobic capacity less than required for the duties they perform. Comparing their standards to those used in some other armies, the authors point out the validity of the same and an imperative to be introduced into Yugoslav Armed forces.

[Evaluation of physical fitness in military personnel using the 1600 meter running test]. / Procena fizicke pripremljenosti pripadnika vojske primenom testa trcanja na 1600 metara.

Validity of 1600 m running test in estimation of physical fitness of Yugoslav Army personnel was examined through the degree of correspondence between the results achieved in the test and the values of maximum oxygen consumption. The coefficients of linear correlation between these two indices are not statistically significant, and amount to -0.36 (in 87 cadets) and -0.14 (in 114 officers). The authors conclude that 1600 m running test is not a valid measure of endurance, as an element of physical fitness of military personnel and they elaborate the reasons of its unreliability.

High incidence of H-ras oncogene mutations in squamous cell carcinoma of lip vermilion.

Nine specimens of well-differentiated squamous cell carcinoma of the lip vermilion have been analyzed for the presence of H-ras oncogene mutations, using the technique of hybridization with synthetic oligonucleotide probes on in vitro amplified tumour DNA. Five specimens harbored mutations: four in codon 12 and one in codon 13. This high incidence (55%) of mutated H-ras genes suggests that their activation may play an important role in lip tumour development and may be connected to the exposure to chemical and/or physical carcinogens.

[Surgical and prosthetic reconstruction of post-tumor defects of the face (case report)]. / Hirursko-proteticka rekonstrukcija posttumorksog defekta lica (Povodom jednog slucaja).

This paper shows a combination of methods in treatment of a facial post tumorous defect. Simultaneous surgical and prosthetic reconstruction was used when treating ablation of the facial basal cell carcinoma in a 70 year old female patient. When planning this surgery the next must be taken into consideration: the age of the patient, general health condition and the size and type of the defect, especially of the bone structures. It means that collaboration between the surgeon and prosthetic is of great importance. In this way a very fast facial rejuvenation can be achieved, even in large defects which enables not only healing but further social rehabilitation of such patients too.

[Immediate prosthetic reconstruction after resection of the upper jaw]. / Imedijatna proteticka rekonstrukcija posle resekcija gornje vilice.

In order to determine the right time when appliance should be applied to the patient after partial, subtotal or total resection of the maxilla, represent cases treated during a six year period at the Clinic of Maxillofacial Surgery of the Stomatology Faculty in Belgrade. In 11 operated patients the appliance was applied immediately after surgery and tamponade of postresection activity cavity. In 46 patients operated at other clinics it was applied more than month after surgery. Thus, 57 patients received immediate prosthetic reconstructive treatment. According to criteria of adaptability considering new appliances major differences could be observed among different groups of patients. 57 patients recovery was normal, while in 4 adaptability was slower. In 12 referral cases not a single case of normal adaptability was registered. 4 patients demonstrated slower adaptability, while in 8 patients severe adaptability problems were registered.

Effects of a modified CMF treatment (cyclophosphamide, methotrexate and 5-fluorouracil) on hematopoietic tissues and Yoshida sarcoma in rats.

Effects of a modified CMF treatment on hematopoietic tissue and an implanted tumor were studied in rats. The modification of the treatment refers to the application of cyclophosphamide 24 h after methotrexate and 5-fluorouracil. The study was done on Wistar rats bearing Yoshida sarcoma in the ascites form. The controls were a) untreated animals bearing the tumor or b) treated conventionally with the 3 cytostatics and c) tumor-free animals under either conventional treatment or d) modified treatment. We examined survival, the appearance of metastases, and the regeneration of hematopoietic tissues. Improved survival, the absence of metastases, and improved regeneration of hematopoietic tissues was observed when modified CMF treatment was applied. These results support the importance of sequencing cytostatic protocols for basic hematological determinants and anti-tumor activity.

Distinct spectrum of N-ras mutations in aml and mds patients in yugoslavia.

Mutations that activate ras genes were demonstrated to be associated with certain types of malignancies. Multiple point mutations were predominantly found in the N-ras and occasionally in the K-ras genes. The analysis of 4 MDS, 23 AML and 11 CML patients from Yugoslavia revealed the prevalence of the N-ras mutation (83%) over K-ras mutations (17%). Although the frequencies of the N- and K-ras mutations in these patients were similar to the ones reported for patients from USA and Japan, the N-ras mutational spectra considerably differed. The prevailing type of mutation in patients from Yugoslavia was G-to-T transversion at the first position in the codon 12 of the N-ras gene. This study supports a hypothesis that different geographical and environmental factors may cause the accumulation of different type of point mutations in the same target gene.