HPV16 E6 Oncogene Contributes to Cancer Immune Evasion by Regulating PD-L1 Expression through a miR-143/HIF-1a Pathway.

Human Papillomaviruses have been associated with the occurrence of cervical cancer, the fourth most common cancer that affects women globally, while 70% of cases are caused by infection with the high-risk types HPV16 and HPV18. The integration of these viruses' oncogenes E6 and E7 into the host's genome affects a multitude of cellular functions and alters the expression of molecules. The aim of this study was to investigate how these oncogenes contribute to the expression of immune system control molecules, using cell lines with integrated HPV16 genome, before and after knocking out E6 viral gene using the CRISPR/Cas9 system, delivered with a lentiviral vector. The molecules studied are the T-cell inactivating protein PD-L1, its transcription factor HIF-1a and the latter's negative regulator, miR-143. According to our results, in the E6 knock out (E6KO) cell lines an increased expression of miR-143 was recorded, while a decrease in the expression of HIF-1a and PD-L1 was exhibited. These findings indicate that E6 protein probably plays a significant role in enabling cervical cancer cells to evade the immune system, while we propose a molecular pathway in cervical cancer, where PD-L1's expression is regulated by E6 protein through a miR-143/HIF-1a axis.

Photo-Fenton and TiO2 Photocatalytic Inactivation of Model Microorganisms under UV-A; Comparative Efficacy and Optimization.

Photocatalytic inactivation of pathogens in aqueous waste is gaining increasing attention. Several homogeneous and heterogeneous photocatalytic protocols exist using the Fenton's reagent and TiO2, respectively. A comprehensive study of homogeneous and heterogeneous photocatalysis on a range of microorganisms will significantly establish the most efficient method. Here, we report a comparative study of TiO2- and Fe+3-based photocatalytic inactivation under UV-A of diverse microorganisms, including Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria, bacterial spores (Bacillus stearothermophilus spores) and viruses (MS2). We also present data on the optimization of TiO2 photocatalysis, including optimal catalyst concentration and H2O2 supplementation. Our results indicate that both photo-Fenton and TiO2 could be successfully applied for the management of microbial loads in liquids. Efficient microorganism inactivation is achieved with homogeneous photocatalysis (7 mg/L Fe+3, 100 mg/L H2O2, UV-A) in a shorter processing time compared to heterogeneous photocatalysis (0.5 g/L TiO2, UV-A), whereas similar or shorter processing is required when heterogenous photocatalysis is performed using microorganism-specific optimized TiO2 concentrations and H2O2 supplementation (100 mg/L); higher H2O2 concentrations further enhance the heterogenous photocatalytic inactivation efficiency. Our study provides a template protocol for the design and further application for large-scale photocatalytic approaches to inactivate pathogens in liquid biomedical waste.

Interaction between TCF7L2 rs7903146 Genotype, HbA1c Levels, and the Periodontal Status of Dental Patients.

OBJECTIVE: The aim of the study was to investigate the potential interaction between TCF7L2 rs7903146 genotype, which is implicated for type-2 diabetes mellitus genetic susceptibility, HbA1c levels, and the periodontal status of dental patients. MATERIALS AND METHODS: HbA1c levels, clinical periodontal parameters (probing depth, clinical attachment level, bleeding on probing, and plaque index), and several parameters (such as body mass index [BMI], smoking habits, education level, and age) were recorded in 150 patients who fulfilled the criteria for screening for prediabetes/diabetes of the Centers for Disease Control and Prevention. DNA was extracted and the TCF7L2 single nucleotide polymorphism (SNP) rs7903146 was genotyped in all participants. RESULTS: Thirty-one patients out of 150 tested were found with unknown hyperglycemia (20.7%). Regarding sex, education, parent with diabetes, normal BMI, smoking, age ≥45 years and prior testing for diabetes, no differences were observed between patients displaying HbA1c < 5.7 and ≥ 5.7% (Pearson's Chi-square test, p > 0.05). Regarding periodontal parameters and differences between subgroups (HbA1c levels ≥ 5.7 and HbA1c levels < 5.7), statistically significant differences were observed for probing depth (3.20 ± 0.94 vs. 2.81 ± 0.78 mm), clinical attachment level (3.54 ± 1.20 vs. 3.18 ± 1.06 mm) and bleeding on probing (0.62 ± 0.25 vs. 0.50 ± 0.24%) with hyperglycemic patients exhibiting worse periodontal conditions (Mann-Whitney test p < 0.05). The allelic and genotype frequencies for the transcription factor 7-like 2 (TCF7L2) gene, SNPs 7903146 did not exhibit a significant difference between the HbA1c > 5.7 and HbA1c < 5.7 groups and the periodontitis and nonperiodontitis subgroups respectively (Fisher's exact test >0.05). Statistical Analysis Patient characteristics and their association with prediabetes were tested by Pearson's Chi-square test (asymptotic, two sided). Differences of periodontal parameters between subgroups were tested with the Mann-Whitney U-test. The associations of allele and genotype frequencies in the patient and control groups were analyzed using the Fisher's exact test of independence.The significance level was set at the 0.05 for all tests. CONCLUSION: A statistically significant association between TCF7L2 rs7903146 genotype and periodontal condition or HbA1c levels was not observed in contrast to statistically significant differences of clinical parameters of periodontitis in patients with hyperglycemia.

Diversity, Cyanotoxin Production, and Bioactivities of Cyanobacteria Isolated from Freshwaters of Greece.

Cyanobacteria are a diverse group of photosynthetic Gram-negative bacteria that produce an array of secondary compounds with selective bioactivity against a broad spectrum of organisms and cell lines. In this study, 29 strains isolated from freshwaters in Greece were classified using a polyphasic approach and assigned to Chroococcales, Synechococcales, and Nostocales, representing 11 genera and 17 taxa. There were good agreements between 16S ribosomal RNA (rRNA)-cpcBA-internal genetic spacer (IGS) characterization and morphological features, except for the Jaaginema-Limnothrix group which appears intermixed and needs further elucidation. Methanol extracts of the strains were analyzed for cyanotoxin production and tested against pathogenic bacteria species and several cancer cell lines. We report for the first time a Nostoc oryzae strain isolated from rice fields capable of producing microcystins (MCs) and a Chlorogloeopsis fritschii strain isolated from the plankton of a lake, suggesting that this species may also occur in freshwater temperate habitats. Strains with very high or identical 16S rRNA gene sequences displayed different antibacterial and cytotoxic activities. Extracts from Synechococcus cf. nidulans showed the most potent antibacterial activity against Staphylococcus aureus, whereas Jaaginema sp. strains exhibited potent cytotoxic activities against human colorectal adenocarcinoma and hepatocellular carcinoma cells. Jaaginema Thessaloniki Aristotle University Microalgae and Cyanobacteria (TAU-MAC) 0110 and 0210 strains caused pronounced changes in the actin network and triggered the formation of numerous lipid droplets in hepatocellular carcinoma and green monkey kidney cells, suggesting oxidative stress and/or mitochondrial damage leading to apoptosis.

The role of Visfatin in atherosclerotic peripheral arterial obstructive disease.

Visfatin is an adipokine molecule acting as an essential coenzyme in multiple cellular redox reactions. The increased serum levels of Visfatin have been correlated with metabolic syndrome and endothelial homeostasis. In this study we investigate the possible relationship of Visfatin serum levels with the severity and location of atherosclerotic peripheral arterial occlusive disease (PAOD). Study protocol included 45 consecutive PAOD and 20 Control patients with age >55years old. Definition of PAOD was based in Rutherord's classification (RC). End-stage PAOD patients (RC-V & -VI) were excluded from study. Data were collected prospectively and included age, gender, atherosclerotic risk factors and the body mass index (BMI). In PAOD patients recorded the PAOD's clinical stage and the presence of carotid stenosis >50%. PAOD patients divided in two subgroups, those with mild (RC-I & -II) and moderate disease (RC-III & -IV). In all serum samples Visfatin was measured, blindly, twice by anosoenzymatic technique. Statistical analysis was performed by non-parametric Mann-Whitney U test, Pearson's chi-square, One Way Anova and Kruskall-Wallis tests, as appropriate. The mean Visfatin value in PAOD and Control groups were 38.5±16.0 and 13.9±3.8ng/ml respectively (p<0.0005). In-PAOD subgroup of patients the visfatin values were not affected by demographics, BMI and atherosclerotic risk factors (p>0.05). Univariate analysis showed that severity of PAOD (mild vs severe), presence of carotid stenosis >50% and multilevel disease significantly affected outcomes (p=0.018, p=0.010 and p=0.006 respectively). In multivariate regression analysis severity of PAOD was the solely factor with strong correlation with high visfatin values (p=0.001). High Visfatin levels seem to be strongly correlated with the presence and severity of PAOD. Further and in depth investigation is needed to define the possible role of Visfatin in atherosclerosis and it's value as a potential prognostic biomarker of PAOD.

Caprine PrP variants harboring Asp-146, His-154 and Gln-211 alleles display reduced convertibility upon interaction with pathogenic murine prion protein in scrapie infected cells.

Scrapie, the prion disease of sheep and goats, is a devastating malady of small ruminants. Due to its infectious nature, epidemic outbreaks may occur in flocks/herds consisting of highly susceptible animals. Field studies identified scrapie-protective caprine PrP variants, harboring specific single amino acid changes (Met-142, Arg-143, Asp-146, Ser-146, His-154, Gln-211 and Lys-222). Their effects are under further evaluation, and aim to determine the most protective allele. We assessed some of these variants (Asp-146, His-154, Gln-211 and Lys-222), after their exogenous expression as murine-caprine chimeras in a scrapie- infected murine cell line. We report that exogenously expressed PrPs undergo conformational conversion upon interaction with the endogenous pathological murine prion protein (PrPSC), which results in the detection of goat-specific and partially PK-resistant moieties. These moieties display a PK-resistance pattern distinct from the one detected in natural goat scrapie cases. Within this cellular model, distinct conformational conversion potentials were assigned to the tested variants. Molecules carrying the Asp-146, His-154 and Gln-211 alleles showed significantly lower conversion levels compared to wild type, confirming their protective effects against scrapie. Although we utilized a heterologous conversion system, this is to our knowledge, the first study of caprine PrP variants in a cellular context of scrapie, that confirms the protective effects of some of the studied alleles.

Prevalence of ß-lactam (bla TEM) and Metronidazole (nim) Resistance Genes in the Oral Cavity of Greek Subjects.

OBJECTIVES: The aim of this study is to investigate the prevalence of bla TEM and nim genes that encode resistance to ß-lactams and nitroimidazoles, respectively, in the oral cavity of systemically healthy Greek subjects. MATERIALS AND METHODOLOGY: After screening 720 potentially eligible subjects, 154 subjects were recruited for the study, including 50 periodontally healthy patients, 52 cases of gingivitis and 52 cases of chronic periodontitis. The clinical parameters were assessed with an automated probe. Various samples were collected from the tongue, first molars and pockets >6mm, and analysed by polymerase chain reaction-amplification of the bla TEM and nim genes, using primers and conditions previously described in the literature. RESULTS: There was a high rate of detection of bla TEM in plaque and tongue samples alike in all periodontal conditions (37% of plaque and 60% of tongue samples, and 71% of participants). The bla TEM gene was detected more frequently in the tongue samples of the periodontally healthy (56%) and chronic periodontitis (62%) groups compared to the plaque samples from the same groups (36% and 29%, respectively; z-test with Bonferroni corrections-tests, P<0.05). The nim gene was not detected in any of the 343 samples analysed. CONCLUSION: The oral cavity of Greek subjects often harbours bla TEM but not nim genes, and therefore the antimicrobial activity of ß-lactams might be compromised.

Prevalence of Staphylococcus aureus and methicillin resistant Staphylococcus aureus (MRSA) in the oral cavity.

OBJECTIVE: To assess the prevalence of Staphylococcus aureus and methicillin resistant Staphylococcus aureus (MRSA) in plaque and tongue samples from systemically healthy subjects with periodontal health, gingivitis or chronic periodontitis. METHODS: After screening 720 potentially eligible subjects, 154 systemically healthy participants were ultimately enrolled in the current study. Subgingival samples were taken from the first molars and the tongue and analyzed for the presence of S. aureus and MRSA by polymerase chain reaction (PCR), using primers and conditions previously described in the literature. In addition, samples were taken from deep periodontal pockets of chronic periodontitis patients. Statistical analysis was performed by applying non-parametric tests (Kruskal-Wallis for clinical parameters, and z-test with Bonferroni corrections for distributions of assessed parameters). All comparisons were set at the 0.05 significance level. RESULTS: S. aureus was detected in 18% of all participants and in 10% of the samples tested. No significant differences were found in its distribution among the three investigated groups (z-test for proportions with Bonferroni corrections, p>0.05). The mecA gene was not present in any of the S. aureus found. CONCLUSIONS: S. aureus can be found in the oral environment regardless of the periodontal conditions and therefore should be considered as a member of the transient flora not participating in periodontal pathology. Subgingival sites and tongue surfaces seem to be an unusual habitat of MRSA.

Superior long-term repopulating capacity of G-CSF+plerixafor-mobilized blood: implications for stem cell gene therapy by studies in the Hbb(th-3) mouse model.

High numbers of genetically modified hematopoietic stem cells (HSCs) equipped with enhanced engrafting potential are required for successful stem cell gene therapy. By using thalassemia as a model, we investigated the functional properties of hematopoietic stem and progenitor cells (HSPCs) from Hbb(th3)/45.2(+) mice after mobilization with G-CSF, plerixafor, or G-CSF+plerixafor and the engraftment kinetics of primed cells after competitive primary and noncompetitive secondary transplantation. G-CSF+plerixafor yielded the highest numbers of HSPCs, while G-CSF+plerixafor-mobilized Hbb(th3)/45.2(+) cells, either unmanipulated or transduced with a reporter vector, achieved faster hematologic reconstitution and higher levels of donor chimerism over all other types of mobilized cells, after competitive transplantation to B6.BoyJ/45.1(+) recipients. The engraftment benefit observed in the G-CSF+plerixafor group was attributed to the more primitive stem cell phenotype of G-CSF+plerixafor-LSK cells, characterized by higher CD150(+)/CD48 expression. Moreover, secondary G-CSF+plerixafor recipients displayed stable or even higher chimerism levels as compared with primary engrafted mice, thus maintaining or further improving engraftment levels over G-CSF- or plerixafor-secondary recipients. Plerixafor-primed cells displayed the lowest competiveness over all other mobilized cells after primary or secondary transplantation, probably because of the higher frequency of more actively proliferating LK cells. Overall, the higher HSC yields, the faster hematological recovery, and the superiority in long-term engraftment indicate G-CSF+plerixafor-mobilized blood as an optimal graft source, not only for thalassemia gene therapy, but also for stem cell gene therapy applications in general.

Perspectives of a scrapie resistance breeding scheme targeting Q211, S146 and K222 caprine PRNP alleles in Greek goats.

The present study investigates the potential use of the scrapie-protective Q211 S146 and K222 caprine PRNP alleles as targets for selective breeding in Greek goats. Genotyping data from a high number of healthy goats with special emphasis on bucks, revealed high frequencies of these alleles, while the estimated probabilities of disease occurrence in animals carrying these alleles were low, suggesting that they can be used for selection. Greek goats represent one of the largest populations in Europe. Thus, the considerations presented here are an example of the expected effect of such a scheme on scrapie occurrence and on stakeholders.

Proteomics for the discovery of biomarkers and diagnosis of periodontitis: a critical review.

Periodontitis is a common chronic and destructive disease whose pathogenetic mechanisms remain unclear. Due to their sensitivity and global scale, proteomics studies offer the opportunity to uncover critical host and pathogen activity indicators and can elucidate clinically applicable biomarkers for improved diagnosis and treatment of the disease. This review summarizes the literature of proteomics studies on periodontitis and comprehensively discusses commonly found candidate biomarkers. Key considerations in the design of an experimental proteomics platform are also outlined. The applicability of protein biomarkers across the progression of periodontitis and unexplored areas of research are highlighted.

Prevalence of antibiotic resistance genes in subjects with successful and failing dental implants. A pilot study.

OBJECTIVES: To investigate the prevalence of the bacterial genes encoding resistance to beta-lactams, tetracyclines and metronidazole respectively, in subjects with successful and failing dental implants and to assess the presence of Staphylococcus aureus and the mecA gene encoding for Methicillin Resistant Staphylococcus aureus (MRSA) in the same samples. MATERIALS AND METHODOLOGY: The subject sample included 20 participants with clinically healthy osseointegrated implants and 20 participants with implants exhibiting peri-implantitis. Clinical parameters were assessed with an automated probe, samples were collected from the peri-implant sulcus or pocket and analyzed with Polymerase Chain Reaction for bla TEM , tetM, tetQ and nim genes, S. aureus and MRSA using primers and conditions previously described in the literature. RESULTS: Findings have shown high frequencies of detection for both groups for the tetracycline resistance genes tetM (>30%), tetQ (>65%) with no statistical differences between them (z-test with Bonferroni corrections, p<0.05). The bla TEM gene, which encodes resistance to beta-lactams, was detected in <15% of the samples. The nim gene, which encodes resistance to metronidazole, S.aureus and the mecA gene encoding for MRSA were not detected in any of the analyzed samples. CONCLUSIONS: Healthy peri-implant sulci and peri-implantitis cases often harbor bacterial genes encoding for resistance to the tetracyclines and less often for beta-lactams. Thus, the antimicrobial activity of the tetracyclines and to a lower extent to beta-lactams, might be compromised for treatment of peri-implantitis. Since no metronidazole resistance genes were detected in the present study, its clinical use is supported by the current findings. S.aureus may not participate in peri-implant pathology.

Prevalence and distribution of Aggregatibacter actinomycetemcomitans serotypes and the JP2 clone in a Greek population.

AIM: to investigate the distribution of Aggregatibacter actinomycetemcomitans serotypes and the prevalence of the JP2 clone in subgingival samples of Greek subjects. MATERIALS AND METHODS: two hundred and twenty eight subjects participated in the present study. Each contributed with one pooled subgingival sample from the mesiobuccal surface of the four first molars. Samples were analysed using polymerase chain reaction for five serotypes of A. actinomycetemcomitans and the JP2 clone, using primers and conditions described previously. Subjects were stratified according to periodontal status (untreated periodontitis, non-periodontitis and periodontitis patients receiving supportive treatment). Comparisons between and within groups were performed by applying non-parametric tests (Kruskall-Wallis, Pearson χ(2) , z-test with Bonferroni's corrections and Kramer's V-test) at p=0.05 level. RESULTS: a. actinomycetemcomitans was detected statistically more frequently in untreated patients (27.5%) compared with the other two groups (11.7% for non-periodontitis and 10% for periodontitis patients receiving supportive treatment). No statistical differences were observed concerning the distribution of serotypes among groups (z-test with Bonferroni's corrections p>0.05). Serotype c was more predominant within the periodontally diseased groups (Kramer's V-test p<0.05). The JP2 clone was not detected. CONCLUSIONS: a. actinomycetemcomitans serotype b was not statistically correlated with periodontal disease in the investigated sample and the utility of microbiological testing before antimicrobial administration is emphasized.

Mobilization of hematopoietic stem cells in a thalassemic mouse model: implications for human gene therapy of thalassemia.

Granulocyte colony-stimulating factor (G-CSF)-mobilized blood stem cells may become the preferable source of hematopoietic stem cells (HSCs) for gene therapy because of the higher yield of cells compared with conventional bone marrow harvesting. A G-CSF-associated risk of splenic rupture has been recognized in normal donors of HSCs, but limited information is available about the G-CSF effect in the presence of splenomegaly and extramedullary hematopoiesis. We investigated the G-CSF effect in a thalassemic mouse model (HBB(th-3)) as compared with a normal strain (C57BL/6), in terms of safety, mobilization efficacy, and distribution of stem cells among hematopoietic compartments. There was no death or clinical sequelae of splenic rupture in G-CSF-treated animals of either strain; however, hemorrhagic infarcts in the spleen were detected with low frequency in G-CSF-treated HBB(th-3) mice (12.5%). HBB(th-3) mice mobilized less effectively than C57BL/6 mice (Lin(-)Sca-1(+)c-Kit(+) cells/microl of peripheral blood mononuclear cells [PBMCs]: 90 +/- 55 vs. 255 +/- 174, respectively, p = 0.01; CFU-GM/ml PBMCs: 390 +/- 262 vs. 1131 +/- 875, p = 0.01) because of increased splenic trapping of hematopoietic stem and progenitor cells (Lin(-)Sca-1(+)c-Kit(+) cells per spleen (x10(5)): 487 +/- 35 vs. 109 +/- 19.6, p = 0.01; CFU-GM per spleen (x10(2)): 1470 +/- 347 vs. 530 +/- 425, p = 0.0006). Splenectomy restored the mobilization proficiency of thalassemic mice at comparable levels to normal mice and resulted in the development of a hematopoietic compensatory mechanism in the thalassemic liver that protected splenectomized mice from severe anemia. Our data imply that, in view of human gene therapy for thalassemia, either multiple cycles or alternative ways of mobilization may be required for a sufficient yield of transplantable HSCs. In addition, strategies to minimize the risk of G-CSF-induced splenic infarcts should be explored in a clinical setting.

Prevalence of tetM, tetQ, nim and bla(TEM) genes in the oral cavities of Greek subjects: a pilot study.

AIM: To investigate the prevalence of tetM, tetQ, nim and bla(TEM) antimicrobial resistance genes in subgingival and tongue samples of Greek subjects. MATERIALS AND METHODS: Fifty-four subjects participated in the present study. Participants each contributed with one pooled subgingival sample from the mesiobuccal surface of the four first molars and one sample from the tongue. Samples were analysed using polymerase chain reaction for tetM, tetQ, nim and bla(TEM) genes using the primers and conditions described previously. Subjects were stratified according to periodontal status (health, gingivitis or periodontitis). Intake of any antibiotic for medical or dental reasons during the previous 12 months was also recorded (self-reported). Comparisons within and between groups were performed by applying non-parametric tests (z-test with Bonferroni corrections). RESULTS: A high prevalence of tetM, tetQ and bla(TEM) genes was detected in both tongue and subgingival samples (48.1-82.2%). No differences were observed across genes between periodontally healthy, gingivitis or periodontitis cases, and no statistical correlation was observed between the presence of the bla(TEM) gene and the intake of beta-lactams during the last 12 months (Fisher's exact test, p>0.05). CONCLUSIONS: Findings from the present study suggest a high prevalence of tetM, tetQ and bla(TEM), but not nim resistance genes in subgingival and tongue samples from Greek subjects.

Human ubiquitin specific protease 31 is a deubiquitinating enzyme implicated in activation of nuclear factor-kappaB.

TRAF2 mediates activation of the transcription factors NF-kappaB and AP1 by TNF. A yeast two-hybrid screen of a human cDNA library identified a ubiquitin specific protease homologue (USP31) as a TRAF2-interacting protein. Two cDNAs encoding for USP31 were identified. One cDNA encodes a 1035-amino acid long isoform of USP31 (USP31, long isoform) and the other a 485-amino acid long isoform of USP31 (USP31S1, short isoform). USP31 and USP31S1 share a common amino terminal region with homology to the catalytic region of known deubiquitinating enzymes. Enzymatic assays demonstrated that USP31 but not USP31S1 possess deubiquitinating activity. Furthermore, it was shown that USP31 has a higher activity towards lysine-63-linked as compared to lysine-48-linked polyubiquitin chains. Overexpression of USP31 in HEK 293T cells inhibited TNFalpha, CD40, LMP1, TRAF2, TRAF6 and IKKbeta-mediated NF-kappaB activation, but did not inhibit Smad-mediated transcription activation. In addition, both USP31 isoforms interact with p65/RelA. Our data support a role for USP31 in the regulation of NF-kappaB activation by members of the TNF receptor superfamily.

Prevalence of IL-1A and IL-1B polymorphisms in a Greek population.

OBJECTIVES: Given the possible ethnic diversity in distribution of genetic variants, aim of the present study was to estimate the prevalence of the polymorphisms of IL-1A+4845 and IL-1B+3954 in a Greek population of unknown periodontal status and to compare this prevalence with one from a group of patients with chronic (adult) periodontitis. MATERIAL AND METHODS: 110 healthy subjects of unknown periodontal status and 45 patients with chronic periodontitis were genotyped, using a PCR-based method and primers described in the literature. The differences in genotype, allele frequencies, allele carriage rate and presence of positive composite genotype as described by Kornman et al. (1997) were analyzed using Fisher's exact test and calculating two-sided p-value, odds ratios and confidence intervals. RESULTS: No differences were observed in any of the parameters tested between the two groups. CONCLUSIONS: Given the high prevalences of these polymorphisms observed in the general population,findings of the present study do not support a possible predictive value of the presence of allele 2 at IL-1A+4845 and IL-1B+3954 or the positive composite genotype for presence or absence of periodontal disease,in a Greek population.

Identification and characterization of the gene products of open reading frame U86/87 of human herpesvirus 6.

The human herpesvirus 6 (HHV-6) immediate early-A locus (IE-A) locates in the position analogous to the human cytomegalovirus (HCMV) major IE (MIE) locus that is well-known to play critical roles in viral infection. Similarly to HCMV MIE, HHV-6 IE-A consists of two genetic units, IE1 and IE2, corresponding to open reading frames U90-U89 and U90-U86/87, respectively. However, the HHV-6 IE-A locus exhibits limited sequence homology with the HCMV MIE locus. In this study, to characterize HHV-6 IE2 gene products, polyclonal antibodies against four domains of the U86/87 open reading frame were generated by immunization of rabbits with bacterially-expressed proteins. Three polypeptides derived from the U86/87 region with apparent molecular masses of 100, 85 and 55 kD were detected in HHV-6-infected cells 3 days after infection, while IE1 polypeptides with apparent molecular mass greater than 170 kD were detectable as early as 8 h. Mapping of the IE2 gene products with the antibodies suggests differential splicing and alternative translation initiation in the IE2 genetic unit. The IE2 products show a mixed cytoplasmic and nuclear localization pattern. In addition, the 437 amino acid carboxyl-terminus domain bound to a DNA fragment containing the putative IE-A promoter. These results suggest that HHV-6 IE2 plays a critical role in transcriptional regulation and viral growth as does HCMV IE2, although it is likely that HHV-6 IE2 has expression kinetics different from HCMV IE2.