Genetic monitoring on the world's first MSC eco-labeled common octopus (O. vulgaris) fishery in western Asturias, Spain.

Octopus vulgaris (Cuvier, 1797) is a cephalopod species with great economic value. In western Asturias (northwest of Spain), O. vulgaris artisanal fisheries are relatively well monitored and conditionally eco-labeled by the Marine Stewardship Council (MSC). Despite this, the Asturian octopus stocks have not been genetically assessed so far. In order to improve the current fishery plan and contrast the octopus eco-label validity in Asturias, 539 individuals from five regions of the O. vulgaris geographic distribution, including temporal samplings in Asturias, were collected and genotyped at thirteen microsatellite loci. All the samples under analysis were in agreement with Hardy-Weinberg expectations. Spatial levels of genetic differentiation were estimated using F-statistics, multidimensional scaling, and Bayesian analyses. Results suggested that the O. vulgaris consists of at least four genetically different stocks coming from two ancestral lineages. In addition, temporal analyses showed stability in terms of genetic variation and high NE (> 50) for several generations in different localities within Asturias, pointing out to indeed sustainable fishery exploitation levels. Even though, the current Asturias fishery plan shows no significant genetic damages to the stocks, the regional-specific management plans need systematic genetic monitoring schemes as part of an efficient and preventive regional fishery regulation strategy.

[Why not revisiting tinidazole as potential treatment of odontogenic infections?]. / Por qué no reevaluar el tinidazol como tratamiento potencial de infecciones odontogénicas?

Tinidazole is a 5-nitroimidazole initially introduced into clinical medicine in 1969 for the treatment of unicellular parasites. Tinidazole offers selective bactericidal activity, not influenced by the inoculum size, against anaerobic bacteria, that make it of theoretical interest against periodontopathogen infections. This article reviews the required characteristics of an antibiotic directed to odontogenic anaerobic infections, as well as the pharmacodynamic pitfalls of common antibiotic treatments. In addition the in vitro, pharmacokinetic and pharmacodynamic properties of tinidazole are reviewed, assessing the degree of its adhesion to the required characteristics, as well as identifying the gaps to be fulfilled prior to its use in this medical field. Tinidazole offers interesting characteristics making worthy investigations as a candidate for the treatment of anaerobic odontogenic infections. \

Odontogenic bacteria in periodontal disease and resistance patterns to common antibiotics used as treatment and prophylaxis in odontology in Spain.

Resistance in streptococci or Gram-negative bacteria is associated with antibiotic consumption. Scarce information exists on the antibiotic susceptibility of bacterial isolates from patients with periodontitis in countries with high antibiotic consumption, as this is an area in which microbiological testing is not performed in daily practice. The present study was undertaken to explore the susceptibility of bacterial isolates in periodontitis to antibiotics prescribed in odontology in Spain as treatment for local infections or prophylaxis for distant focal infections. Periodontal samples were prospectively collected in 48 patients classified by pocket depth of <4 mm and >or=4 mm. Species were identified by culture, selecting the five most frequent morphotypes per sample, and polymerase chain reaction (PCR). Susceptibility was determined by E-test. A total of 261 isolates were identified: 72.9% patients had Streptococcus oralis; 70.8% Streptococcus mitis; 60.4% Prevotella buccae; 39.6% Prevotella denticola; 37.5% Fusobacterium nucleatum; 35.4% Prevotella intermedia; 25% Capnocytophaga spp.; 23% Veillonella spp.; 22.9% Prevotella melaninogenica and Streptococcus sanguis; and <20% other species. Streptococcus viridans resistance rates were 0% for amoxicillin, approximately 10% for clindamycin, 9-22% for tetracycline, and for azithromycin ranged from 18.2% for S. sanguis to 47.7% for S. mitis. Prevotella isolates were susceptible to amoxicillin-clavulanic acid, with amoxicillin resistance ranging from 17.1% in P. buccae to 26.3% in P. denticola. Metronidazole resistance was <6% in all Prevotella species, while clindamycin resistance ranged from 0 to 21.1%. beta-Lactamase production was positive in 54.1% Prevotella spp., 38.9% F. nucleatum, 30% Capnocytophaga spp., and 10% Veillonella spp. In this study, amoxicillin-clavulanic acid was the most active antibiotic against all species tested, followed by metronidazole in the case of anaerobes.

Oral squamous cell carcinoma and biological markers: an update on the molecules mainly involved in oral carcinogenesis.

Squamous cell carcinoma (SCC) is one of the most common malignant cancer of the oral cavity encompassing at least 92.8% of all oral malignancies. Despite improved diagnostic and therapeutic methods over the 20 last years, this tumour is still characterized by a high rate of mortality. The latest advances of molecular biological methods have contributed to better understand the mechanisms involved in the oral carcinogenetic process. Deregulation of cell cycle, apoptosis and cell-cell/cell-matrix adhesions are considered the pathways mainly influencing this multistage event and scientific researches over the last decade have been performed in order to investigate the biological diagnostic and prognostic parameters related to these events (i.e. tumour growth markers, markers of tumour suppression and anti-tumour response, angiogenesis markers, markers of tumour invasion and metastatic potential, cell surface markers, intracellular markers, markers derived from arachidonic acid, and enzymatic markers). The aim of the present review was to outline the current knowledge on the role of some of these tumour biological markers in carcinogenesis of oral SCC.

Apoptosis and cell cycle arrest in oral lichen planus Hypothesis on their possible influence on its malignant transformation.

The quantitative importance of cell cycle arrest and apoptosis mechanisms in oral lichen planus (OLP) was analysed in order to assess the cell response to T lymphocyte aggression and establish a hypothesis on the influence of these phenomena in the malignant transformation process. The TUNEL assay and immunohistochemical methods were used to detect caspase-3, bax, and p21 in 32 tissue samples of oral mucosa with OLP and in 20 samples of normal oral mucosa. Positivity for TUNEL, caspase-3 and p21 was significantly more frequent in cases than in controls (p<0.001). Both TUNEL and caspase-3 positivity was significantly greater in the basal versus suprabasal layer (p=0.004 and 0.052, respectively). The basal and suprabasal expression of p21 was significantly higher in cases with a more intense liquefaction degeneration (p<0.01). There was no significant difference in basal expression of bax between cases and controls. The quantitative importance of apoptosis was small in OLP. Epithelial cells attacked in OLP have a very low response to apoptosis and cell cycle arrest mechanisms, which may produce an epithelial substrate that favours malignant transformation.

[From knowledge of bacterial etiology to the treatment and prevention of the most prevalent infections in the community: odontological infections]. / Del conocimiento de la etiologia bacteriana al tratamiento y la prevencion de las infecciones mas prevalentes en la comunidad: las infecciones odontologicas.

The prevalence of community odontological infections and their relevance to public health were reviewed. Knowledge of bacterial etiology (and the transmittability of these microbial agents) was used to study the disease (individual susceptibility to etiological agents) in order to review the effect of treatment on odontological pathogens and human microbials. The synergy between the primary care physician and the dentist is fundamental to the control of this endemic disease.

Implantation of sepiolite-collagen complexes in surgically created rat calvaria defects.

The response of osseous tissue to the implantation of sepiolite-collagen complexes has been studied. Sepiolite, sepiolite-collagen complex and 0.5% glutaraldehyde-treated sepiolite-collagen complex were implanted in created circular defects in rat calvaria. The tissue reactions were analysed using light, transmission and scanning electron microscopies. The patterns of bone growth were radiographically analysed and the bone activity was indirectly quantified by using a point-count method. The reaction against the three implanted materials is characteristic of a foreign body reaction with abundant macrophages and giant cells. Implanted products have been detected in macrophages, which suggest the involvement of phagocytosis in the resorptive process. Bone grew at the implantation sites originating excrescences or sometimes a thin bridge at the defect margins. The studied materials, after implantation in contact with bone tissue, did not produce any toxic effect or necrosis, allowing bone activity.

Flow cytometric DNA ploidy analysis of oral cancer comparison with histologic grading.

It has been reported that DNA content analysis provides prognostic information eliminating the subjective component involved in routine microscopic decision making. In an attempt to establish a relationship between pathological findings and flow cytometric analysis, 36 formalin-fixed, paraffin-embedded tumour tissue samples were prepared according to Hedley's method and analysed by means of an Epics Profile II flow cytometer. DNA aneuploidy was observed in 15 tumours (41%). A statistically significant correlation was identified between DNA index and mitoses, cellular response and degree of differentiation, but not the ploidy status. It was not possible to identify a significant association of sex, age, and site of the tumour to DNA index. We found a strong correlation between histologic malignancy and DNA index; an increase in DNA index as malignancy score increases was noted. It was concluded that DNA index shows a good correlation with the histologic features of oral cancer, being a complement of differentiation and histologic grading analysis. The use of DNA analysis as a complement to pathological studies would help to diminish the subjective component of assessment of head and neck cancers. Ploidy status was not statistically associated with the differentiation of tumours.

P21WAF1/CIP1 protein and tongue cancer prognosis.

The expression of the cell cycle regulatory protein p21waf1/cip1 was analyzed immunohistochemically in tissue from the tumor and adjacent non-tumor epithelia of 54 patients with tongue cancer in order to determine the proportion of tongue carcinomas with altered p21waf1/cip1 expression, establish whether this alteration is an early event in tongue carcinogenesis and to investigate whether p21waf1/cip1 expression has predictive prognostic value in these tumors. The percentage of p21waf1/cip1-positive neoplastic cells was calculated. Adjacent non-tumor epithelium was classified as normal, hyperplastic or dysplastic, and the p21waf1/cip1 expression was only considered normal in suprabasal layers. P21waf1/cip1 expression was negative in 57.4% (31/54) of the tumors and was aberrant in the non-tumor adjacent tissue of all patients studied. Neither the absence nor the degree of p21waf1/cip1 expression influenced the survival of patients in the present series. P21waf1/cip1 system alteration may be an early and frequent event in tongue carcinogenesis.

Metastatic lung carcinoma involving the periodontium. Report of a case.

A case of metastatic bronchogenic carcinoma to the gingiva in a 47-year-old male is reported. The gingival lesion developed as a quickly growing mass and appeared 2 months after surgical excision and radiotherapy of the lung carcinoma were completed. The gingival tumor was histopathologically diagnosed as a poorly differentiated squamous carcinoma. Comparative cytologic studies showed similarities between the gingival metastasis and the previous lung cancer.

Levels of interleukin-1 beta, -8, and -10 and RANTES in gingival crevicular fluid and cell populations in adult periodontitis patients and the effect of periodontal treatment.

BACKGROUND: Various cytokines have been identified at sites of chronic inflammation such as periodontitis. Cytokines are synthesized in response to bacteria and their products, inducing and maintaining an inflammatory response in the periodontium. The purpose of the present study was to investigate the involvement of interleukin-1 beta (IL-1 beta), IL-8, and IL-10 and RANTES (regulated on activation, normally T cell expressed and secreted) and the cell populations associated with the immune response in destructive periodontitis, as well as the effect of periodontal therapy on cytokine levels in gingival crevicular fluid (GCF). METHODS: Data were obtained from 12 patients with moderate to advanced periodontitis and 6 healthy controls. Patients presenting at least 2 sites with > or =2 mm clinical attachment loss were included in the destructive periodontitis group. After monitoring for 4 months, only 6 patients showed destructive periodontitis and GCF samples and soft tissues biopsies were collected from these patients. GCF samples and biopsies were collected both from active (12 CGF samples and 6 biopsies) and inactive (12 CGF samples and 6 biopsies) sites. The comparison with healthy controls was carried out by collecting GCF samples from 6 healthy volunteers (12 samples) and biopsies during the surgical removal of wisdom teeth. In periodontal patients, clinical data and GCF samples were obtained prior to periodontal treatment (72 samples) and 2 months after periodontal therapy (72 samples). GCF was collected using a paper strip; eluted and enzyme-linked immunoabsorbent assays (ELISA) were performed to determine cytokine levels. The inflammatory infiltrate was analyzed by immunohistochemistry of gingival biopsy samples with monoclonal antibodies against CD3, CD8, CD4, CD11c, and CD19 antigens. RESULTS: Cellular components of the inflammatory infiltrate include B and T lymphocytes and monocyte/macrophages. Active sites contained a higher number of B lymphocytes and macrophages. IL-8 and IL-1 beta and RANTES in GCF were detected in the majority of sites from periodontal patients (100%, 94% and 87%, respectively); IL-10 was found in only 43%. IL-8 was the only cytokine detected in the GCF (75%) of the control group. Moreover, IL-1 beta levels were significantly higher in active sites versus inactive sites (P <0.05). IL-8 and IL-10 and RANTES were increased in active sites; however, differences were not significant (P>0.05). A positive correlation between the IL-8 and RANTES (r = 0.677, P<0.05) was observed in periodontitis patients. Periodontal therapy reduced the total amount of IL-1 beta, IL-8, and IL-10 and RANTES. Data showed a weak correlation between the clinical parameters and the total amount of cytokines in periodontitis. CONCLUSIONS: These data suggest that the amount of crevicular IL-1 beta, IL-8, and IL-10 and RANTES is associated with periodontal status. Removal of the bacterial plaque reduces the antigenic stimuli and consequently could modulate the chemokines present in GCF. We propose that the dynamic interactions between cytokines, their production rates, and their quantity could represent factors controlling the induction, perpetuation, and collapse of the cytokine network present in the periodontal disease.

Apoptosis in chronic adult periodontitis analyzed by in situ DNA breaks, electron microscopy, and immunohistochemistry.

BACKGROUND: Apoptosis is an evolutionary form of physiological cell death. Previous studies suggest that apoptosis is involved in the pathogenesis of periodontal diseases. Therefore, we studied the apoptotic events in the gingival tissue of chronic adult periodontitis patients. METHODS: Gingival tissue biopsies from 22 patients with chronic adult periodontitis and from 11 healthy controls were obtained. Criteria for patient inclusion in the periodontitis group were a minimum of 14 natural teeth, excluding third molars, with at least 10 posterior teeth; 5 to 6 sites with probing depth > or = 5 mm; attachment loss > or = 3 mm; and extensive radiographic bone loss. The control group included healthy subjects with no prior history of periodontal disease. Apoptosis was determined using the terminal TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique; electron microscopic analysis; and expression of Caspase-3, Fas, FasL, Bcl-2, and p53 by immunohistochemistry. RESULTS: TUNEL-positive cells and cells exhibiting chromatin condensation by electron microscopy were observed in the inflammatory infiltrate of biopsies obtained from periodontitis patients. Most of the TUNEL-positive cells belonged to neutrophil cell populations as they were stained with anti-myeloperoxidase. Positive staining for active-caspase 3, Fas, FasL, and p53 was only observed in the inflammatory infiltrate from periodontitis biopsies, whereas Bcl-2 cells were present in both periodontitis patients and healthy controls. CONCLUSIONS: Our findings establish that apoptosis is induced in the periodontal tissue by host and microbial factors and support the hypothesis that apoptotic mechanisms could be implicated in the inflammatory process associated with gingival tissue destruction observed in adult periodontitis patients.

Tinidazole inhibitory and cidal activity against anaerobic periodontal pathogens.

The in vitro activity of tinidazole against anaerobic periodontal pathogens (25 Prevotella buccae, 18 Prevotella denticola, 10 Prevotella intermedia, 6 Prevotella melaninogenica, 5 Prevotella oralis, 10 Fusobacterium nucleatum and 8 Veillonella spp.) was determined by agar dilution. MIC(90) values (minimum inhibitory concentration for 90% of the organisms) were 8 microg/mL for Veillonella spp., 4 microg/mL for P. intermedia, 2 microg/mL for P. buccae, 1 microg/mL for Fusobacterium spp. and 0.5 microg/mL for other Prevotella spp. Cidal activity was studied by killing curves with tinidazole and amoxicillin (alone and in combination) at concentrations similar to those achieved in crevicular fluid (41.2 microg/mL tinidazole and 14.05 microg/mL amoxicillin) against an inoculum of ca. 10(7)colony-forming units/mL of four bacterial groups, each one composed of four different strains of the following periodontal isolates: Prevotella spp., Fusobacterium spp. and Veillonella spp. (anaerobes) and one amoxicillin-susceptible Streptococcus spp. (facultative) in a proportion of 1:1:1:1. When only beta-lactamase-negative Prevotella or Fusobacterium strains were tested, significantly higher reductions were found with amoxicillin (>4 log reduction at 48 h) versus controls. The presence of beta-lactamase-positive Prevotella spp. or F. nucleatum strains rendered amoxicillin inactive (no reductions at 48 h), with no differences from controls. Amoxicillin+tinidazole produced >3 log reduction at 24h and >4 log reduction at 48 h regardless of the presence or not of beta-lactamase-positive strains. The presence in crevicular fluid of beta-lactamases produced by beta-lactamase-positive periodontal pathogens may have ecological and therapeutic consequences since it may protect beta-lactamase-negative periodontal pathogens from amoxicillin treatment. In vitro, tinidazole offered high antianaerobic activity against beta-lactamase-positive and -negative periodontal pathogens, avoiding amoxicillin inactivation.

Oral complication risks after invasive and non-invasive dental procedures in HIV-positive patients.

BACKGROUND: Limited published scientific evidence is available to provide guidance to clinicians on possible increased risks of invasive oral procedures associated with the human immunodeficiency virus (HIV) status of the patient. The aim of this study was to assess post-procedural complications in patients infected with HIV. MATERIAL AND METHODS: This was a retrospective cross-sectional study of the records of 101 consecutive HIV patients treated at the School of Dentistry of Madrid Complutense University and Sandoval STD Clinic in Madrid between January 2003 and February 2005. Data were gathered by an experienced dental practitioner using a structured epidemiological questionnaire for information on gender, age, HIV transmission category, medical history, hepatitis B virus (HBV) or hepatitis C virus (HCV) coinfection and other diseases, TCD4+ and TCD8+ count, HIV viral load (VL), platelet count, neutrophil count, international normalized ratio and haemoglobin level; tobacco and alcohol intake, highly active antiretroviral treatment and presence of oral lesions. Information was also collected on complications related to dental treatment (invasive or non-invasive) during the previous 6 months. Chi-squared test and Fisher's exact test were used to establish statistical significance. RESULTS: Data were gathered on 314 dental procedures in 101 patients. The overall complication rate was 2.2% (7/314); in 147 invasive procedures, seven complications (4.8%) were documented (one persistent pain, two prolonged bleeding, three infections, one bone sequestrum) including extractions, periodontal scaling, endodontic treatment and biopsy. No differences were found in TCD4+, TCD8+, platelet count, HBV or HCV co-infections or HIV VL between patients with and/or without complications. Patients with complications were mainly in B stage of HIV disease (P=0.020). Oral lesions and smoking habit>20 cig day-1 were documented in 83.3% (P=0.086) and 50% (P=0.060), respectively, of patients with complications. CONCLUSIONS: The complication rate was 2.2% overall and 4.8% after invasive dental procedures. Presence of oral lesions, smoking habit or HIV clinical stage B may be predictive factors for oral complications in HIV patients. No relationship was found between complications and virological, immunological or other laboratory values. Studies with wider samples and negative control group are warranted to confirm the absence of an association between HIV positivity and higher risk of oral complications.

Oral transmission of HIV, reality or fiction? An update.

Human immunodeficiency virus (HIV) and many other viruses can be isolated in blood and body fluids, including saliva, and can be transmitted by genital-genital and especially anal-genital sexual activity. The risk of transmission of HIV via oral sexual practices is very low. Unlike other mucosal areas of the body, the oral cavity appears to be an extremely uncommon transmission route for HIV. We present a review of available evidence on the oral-genital transmission of HIV and analyse the factors that act to protect oral tissues from infection, thereby reducing the risk of HIV transmission by oral sex. Among these factors we highlight the levels of HIV RNA in saliva, presence of fewer CD4+ target cells, presence of IgA antibodies in saliva, presence of other infections in the oral cavity and the endogenous salivary antiviral factors lysozyme, defensins, thrombospondin and secretory leucocyte protease inhibitor (SLPI).

Chemokine RANTES in gingival crevicular fluid of adult patients with periodontitis.

BACKGROUND, AIMS: This study presents the first evidence on the presence of the chemokine RANTES in the gingival fluid crevicular (GCF) of patients with periodontitis. RANTES is a chemokine that selectively attracts and activates macrophages and lymphocytes. Leucocytes play a critical rôle in the host response to the subgingival microflora. METHOD: In this study, the presence de RANTES in GCF was determined in samples obtained from adult patients with periodontitis and from control subjects with clinically healthy gingiva. GCF was collected from different probing depths (<3 mm, 4-6 mm, >6 mm) (n = 72); and active (n = 12) and inactive sites (n = 12). An active site was defined as attachment loss >2 mm, as determined by sequential probing and the tolerance method. GFC was collected for 30 s using Periopaper strips, and RANTES was quantified by ELISA. RESULTS: The presence of RANTES was detected exclusively in the group of patients with periodontitis, presenting a total amount of 40.43+/-16 pg and a concentration 67.80+/-41 pg/microl. RANTES concentration was significantly higher in probing depth <3 mm than in probing depth >6 mm (87.24 versus 51.87, p = 0.014). Total amount and concentration in the GCF samples from active sites were higher that in inactive sites (p>0.05). CONCLUSIONS: The finding that RANTES is found only in patients with periodontitis, may represent a general feature of chronic inflammatory in periodontal diseases. Finally, RANTES may be implicated in the biological mechanisms underlying the pathogenesis and progression of periodontal disease.

Oral candidiasis as a clinical marker related to viral load, CD4 lymphocyte count and CD4 lymphocyte percentage in HIV-infected patients.

BACKGROUND: High viral load is currently considered to be one of the main indicators of the progression of HIV-induced immunodepression, but few studies have analysed its relationship to the presence of oral candidiasis (OC). The aim of this cross-sectional study is to analyse the relationship between viral load, total CD4 lymphocyte count, and percentage of CD4 lymphocytes to the occurrence of OC. METHODS: The present cross-sectional study included 156 HIV-infected patients seen at a clinic for sexually transmitted diseases and HIV. We assessed the presence or absence of OC, and microbiological samples were obtained from the palatine mucosa and dorsal tongue for a smear stained with KOH (potassium hydroxide) and culture on Sabouraud's dextrose agar in all patients. Viral load was determined by quantification of viral RNA in peripheral blood with a minimum detectable level of 500 RNA copies/ml. CD4+ counts/CD4+ percentage were categorized as <200/<14%, 200-499/14-28%, and >500/>29%, and HIV viral loads were categorized as <500, 500-10,000, >10,000 copies/ml. RESULTS: Thirty-eight percent (37.8%) of the patients had OC. Patients with CD4+ lymphocyte counts below 200 x 10(6)/l and CD4+ percentages below 14% showed a significantly higher frequency of OC (57.9% and 48.0%, respectively). Patients with a viral load over 10,000 copies/ml also had OC more frequently (44.8%). In the multiple logistic regression analysis, OC showed a statistically significant association with high viral load [>10,000 vs <500, odds ratio (OR)=11.4], low percentage of CD4+ lymphocytes (<14% vs >28%, OR=5), and injection drug use (IDU vs heterosexual transmission, OR=10.2). In HIV-infected patients, high viral load was associated with more frequent OC, regardless of CD4+ lymphocyte level. CONCLUSIONS: These findings suggest that oral candidiasis could be a useful clinical marker of patients with high viral load. In view of these results, emphasis should be placed on the importance of systematic examination of the oral cavity in all medical follow-up examinations of HIV-infected patients.

[Conceptual review of Behçet's syndrome. Diagnostic and therapeutic criteria]. / Revisión conceptual del sindrome de Behçet. Criterios diagnósticos yterapéuticos.

A review of the literature on clinical, diagnostic and therapeutic aspects on Behcet Syndrom is done.