L1 (CAM) (CD171) in ovarian serous neoplasms.

PURPOSE OF THE INVESTIGATION: The evaluation of L1 (CAM) as a tumor progression marker and as a prognostic factor in serous ovarian tumors. METHODS: L1 (CAM) protein expression was assessed by immunohistochemistry and Western blot in serous ovarian tumors [cystadenomas (n = 20), borderline tumors (n = 14) and carcinomas (n = 47)], and was correlated with stage,grade, progression-free survival time (PFS) and overall survival. RESULTS: L1 (CAM) immunoreactivity correlated significantly with stage and grade. It increased from benign tumors to early carcinomas and to advanced stage carcinomas progressively and significantly. In Stage III G3 carcinoma patients, low L1 (CAM) expressing tumors exhibited better response to chemotherapy and were associated with statistically significantly longer PFS (p = 0.002). CONCLUSION: L1 (CAM) expression represents a novel diagnostic marker in serous ovarian neoplasms that shows characteristics of tumor progression. L1 expression was associated with chemotherapy response.

Evaluation of pupil mobility in patients with myasthenia gravis.

The aim of this study is to investigate the effect of Myasthenia Gravis (MG) on the Central Nervous System (CNS) and/or the smooth muscles of the iris through pupillometry. Sixteen recently diagnosed Myasthenic and sixteen non-Myasthenic subjects of matching age and gender underwent a pupillometric study of the effects of single flash stimuli of 24.6 candelas/m2 intensity and 20 msec duration. A significant decrease in Amplitude (p < 0.001), Maximum Constriction Velocity (p < 0.001) and especially Maximum Constriction Acceleration with a perfect discrimination ability (AUC= 1, p < 0.001). was observed in the Myasthenic compared to the non-Myasthenic subjects. In contrast, no significant difference was observed in Baseline Pupil Radius (R1) and 3.5 secs Percentage Recovery-Redilatation (R%) (p = 0.051 and p = 0.517, respectively). Of the parameters that are studied, R1 and R% are governed mainly by the action of the Sympathetic Nervous System (SNS) and the rest by the Parasympathetic Nervous System (ParNS), through Acetylcholine. The analysis of these parameters demonstrates that the SNS remains unaltered while the ParNS may be affected in MG. This post-synaptic cholinergic receptors' deficit may be central, within the CNS, or peripheral, related to the Neuromuscular Junction of the iris' sphincter.

Retinoid receptor alpha and Beta expression in serous ovarian tumors.

The expression of retinoid acid receptors alpha (RARalpha) and beta (RARbeta) and estrogen receptor alpha (ERalpha) was assessed by immunohistochemistry and Western blotting in normal ovaries, serous cystadenoma (n = 20), serous borderline (n = 14), and serous ovarian cancer (n = 47) and was correlated in cancer cases with stage, grade, progress-free survival (PFS), and survival. RARalpha was increasingly expressed in benign cystadenomas, borderline, and low-stage and advanced-stage neoplasms (p < 0.001). In stage III, G3 serous carcinoma, increased RARalpha expression was an independent prognostic factor associated with lower chemoresponse to first-line chemotherapy (taxol and carboplatin) and shorter PFS (p < 0.002).RARbeta and ERalpha expression did not correlate with RARalpha tumor characteristics or PFS and survival.

Contamination of a toothbrush with antibacterial properties by oral microorganisms.

OBJECTIVES: The aim of this study was to examine the contamination and the survival rate of periodontopathic and cariogenic species on new toothbrushes with antibacterial properties (coated bristles with triclosan), after a single use in periodontitis patients. The decontamination effect of the use of toothpaste was also evaluated. METHODS: Ten patients, who consulted the Department of Periodontology, for treatment of chronic periodontitis, were selected. In each patient four different toothbrushes were used. Two quadrants, randomly selected, were each brushed using a different antibacterial toothbrush. In one of these two quadrants toothpaste was used. The same happened with the remaining quadrants, only with regular toothbrushes. After brushing, the toothbrushes were rinsed and stored in room temperature and a dry environment. After 0, 4 and 24h, four tufts, from each toothbrush, were cut and processed for selective and non-selective culturing techniques, followed by identification and quantification of all species found. RESULTS: Immediately after brushing the toothbrushes harbored a significant number of microorganisms, with no statistically significant difference between the two types of brushes (regular and antibacterial). The reduction of microorganisms from 0 to 4h after brushing was statistically significant (p<0.05). The difference was less obvious from 4 to 24h. When toothpaste was used, brushes harbored significantly (p<0.05) lower numbers of colony-forming units (CFU) compared to those without the use of toothpaste. CONCLUSIONS: The antibacterial toothbrush with triclosan coated tufts failed to limit the bacterial contamination. The toothpaste, on the other hand, significantly reduced the contamination of toothbrushes.

Effect of age on pupillary light reflex: evaluation of pupil mobility for clinical practice and research.

PURPOSE: The aim of this study was to provide a data base for the measurement of various parameters of the pupil light reflex in normal subjects using a fast video pupillometry device (262 frames/sec). METHOD: One hundred healthy subjects took part in the study aged 44.31+/-18.11 years. Subjects were divided in two (2) groups according to age: 18-50 years of age (group 1) and 51-81 years of age (group 2). All subjects were examined between 09.00 and 15.00 and the re-test examination was repeated exactly 24 hours later. All variables showed satisfactory test-retest reliability (Pearson test-retest showed values over 0. 70 for all parameters besides Time for Maximum Constriction (T3. 0.62) and Recovery (R%: 0.57)). RESULTS: The Latency of pupil reaction to light (Ti) was not affected by Age. Baseline Pupil Radius (RI) after 2 min of dark adaptation was statistically smaller in the elderly group p < 0.001; Maximum Constriction Velocity (VCmax), Maximum Constriction Acceleration (ACmax) and Amplitude (AMP) were significantly smaller in the elderly group (p < 0.001) when compared to group 1. When all parameters were studied their correlation showed a statistical significant difference for R1, R2, VCmax, ACmax and AMP when related to Age. However when Age was taken into account through the use of partial correlation, the relation between R1, R2 and AMP remained unaltered, but the relations between R1 and VCmax and ACmax were dramatically reduced from -0.39 to -0.21 and from -0.45 to -0.09 respectively indicating that the relation observed between Ri and VCmax and ACmax was due mainly to the Age of the subjects. CONCLUSION: The results suggest that age influences Baseline Pupil Size, Maximum Constriction Velocity (VCmax) and Acceleration (ACmax), while the Latency of the light reflex remains unaltered.

Early microbial colonization of permucosal implants in edentulous patients.

In edentulous patients, the microbial colonization of permucosal implants of sintered hydroxyapatite was studied. Samples were taken from mucosa and dentures before insertion of implants and from supra- and subgingival sites two to 10 weeks after insertion. In total, five patients and 10 implants with clinically healthy peri-implant tissues were studied. The samples were investigated by dark-field microscopy and anaerobic culture. The supragingival plaque of the implants was dominated by Gram-positive cocci and rods, the subgingival plaque by Haemophilus spp. and Veillonella parvula. A group of bacteria was found specifically related to the implants: Actinomyces odontolyticus, Peptostreptococcus micros, Haemophilus actinomycetemcomitans, Eikenella corrodens, Capnocytophaga sputigena, and Leptotrichia buccalis. Black-pigmented Bacteroides was not found in any of the examined samples. Spirochetes were observed in denture plaque samples and in supragingival plaque of the implants. It is concluded that bacteria known as potential periodontal pathogens colonize the permucosal implants in the first weeks after insertion. The presence of these species seems to be dependent on the ecological factors provided by the artificial gingival crevice of the permucosal implants in the edentulous mouth.

The clinical and microbiological effects of systemic ornidazole in sites with and without subgingival debridement in early-onset periodontitis patients.

BACKGROUND: The purpose of this study was to evaluate the clinical and microbiological effects of systemic ornidazole (ORN) in sites with or without subgingival debridement in early-onset periodontitis (EOP) patients. METHODS: Two pooled bacterial samples consisting of 4 sites each (scaled and non-scaled sites) were obtained from 30 individuals exhibiting EOP. All patients received oral hygiene instruction (OHI), supragingival scaling and ORN. Subgingival scaling and root planing (SRP) was carried out only in scaled sites. Bacterial samples were taken at baseline (BL) and 1 week and 2, 6, and 12 months after systemic ornidazole administration (500 mg/bid for 7 days). One more sample was taken at scaled sites, one week after SRP. RESULTS: One week following SRP (scaled sites) Gram-negative facultative and anaerobic rods were significantly reduced while Gram-positive facultative cocci were significantly increased. After ORN administration, P. gingivalis, P. denticola, P. intermedia, B. forsythus, C. rectus, and S. sputigena were no longer detectable in either scaled or non-scaled sites. A statistically significant long-term (2, 6, and 12 months) reduction of P. gingivalis, P. intermedia, P. loescheii, B. forsythus, and C. rectus and a pronounced increase of S. milleri, S. oralis, and S. sanguis counts in both scaled and non-scaled sites were detected in comparison to baseline. A sustained reduction of bleeding tendency and of probing depth was also observed in both scaled and non-scaled sites. CONCLUSIONS: ORN combined with SRP effects beneficial shifts in the bacterial population associated with substantial clinical improvement, thereby indicating that ORN is effective adjunct in the treatment of EOP deep periodontal pockets where anaerobic bacteria are predominant.

Microbiota of rapidly progressive periodontitis lesions in association with clinical parameters.

The microbial population in 73 rapidly progressive periodontitis (RPP) lesions in 10 young adults aged 25 to 35 years (5 males, 5 females) was studied in relation to the clinical parameters probing depth, bleeding on probing, and suppuration, which were recorded at the sampled sites. Porphyromonas gingivalis was found to predominate (26.7%) in 73 periodontal lesions with clinical probing depth > 6 mm, followed by Bacteroides forsythus (23.6%), and Prevotella intermedia (15.7%). The prevalence of P. gingivalis was 91.7%, Fusobacterium nucleatum, 90.4%; Streptococcus intermedius, 87.7%; and B. forsythus, 53.4%. Significant differences between bleeding index 0, 1, and 2 (P < 0.05) in frequency of detection were found for P. intermedia, Campylobacter concisus, Selenomonas sputigena, and Peptostreptococcus micros at bleeding sites and for Streptococcus sanguis, Actinobacillus actinomycetemcomitans, and B. forsythus (P < 0.001) at non-bleeding sites. Between suppurating and non-suppurating sites, significant differences (P < 0.05) in frequency of isolation were found for P. intermedia, Capnocytophaga ochracea, and A. actinomycetemcomitans at suppurating sites and for F. nucleatum at non-suppurating sites.

Subgingival microflora associated with nifedipine-induced gingival overgrowth.

The purpose of this study was to examine the composition of subgingival plaque of 140 periodontal lesions in 35 patients with cardiovascular disorders who were administered nifedipine and manifested nifedipine-induced gingival overgrowth (GO). Age was inversely associated with the GO. Plaque index and bleeding index showed a significant association with GO, while nifedipine dosage and duration of nifedipine therapy were not found to be significant predictors of GO. The gingival inflammation as expressed in the logistic regression model by the interaction term color x tone was found to be significantly associated with the GO. Statistically significant differences between the groups of comparable probing depth and different degrees of GO were detected for Propionibacterium acnes, Capnocytophaga gingivalis, Capnocytophaga ochracea, Capnocytophaga sputigena, Bacteroides gracilis, Fusobacterium mortiferum, Fusobacterium nucleatum, Fusobacterium varium and Selenomonas sputigena in deep and enlarged lesions. Significantly more frequently isolated were the bacterial species Eubacterium alactolyticum, Campylobacter concisus, C. gingivalis, C. ochracea, C. sputigena, F. mortiferum, F. nucleatum, and F. varium from the more enlarged lesions (GO >3).

Cholinergic deficiency in Alzheimer's and Parkinson's disease: evaluation with pupillometry.

The aim of the study was to evaluate the cholinergic deficiency in Alzheimer's (AD) and Parkinson's disease (PD). For this purpose, pupil size changes and mobility were assessed using a fast-video pupillometer (263 frames/s). Twenty-three (23) patients with probable AD and twenty-two (22) patients with PD (eleven with cognitive impairment and eleven without) entered the study. A full record of the pupil's reaction to light was registered. From this data ten (10) parameters were measured and reported. Comparison of those parameters in both group of subjects followed. Patients with probable AD had abnormal pupillary function compared to healthy ageing. All the Pupil Light Reflex (PLR) variables significantly differed between the two groups (p<0.005) except the Baseline Pupil Diameter after 2-min dark adaptation (D1) and the Minimum Pupil Diameter (D2). Maximum Constriction Acceleration (ACmax) was the best predictor in classifying a subject as normal or as an AD with a perfect classification ability (AUC=1, p<0.001). ACmax and Maximum Constriction Velocity (VCmax) were significantly lower in PD patients without and with coexisting cognitive impairment compared to normal subjects (p<0.001). Patients with cognitive impairment had significantly lower levels of ACmax, VCmax and amplitude (AMP=D1-D2) than patients with no cognitive deficits. ACmax and secondarily VCmax were the best predictors in classifying a subject as normal or as a PD patient with or without cognitive impairment. Cognitive and memory impairment, which reflects a cholinergic deficit, may be a crucial pathogenetic factor for the decrease in the aforementioned pupillometric parameters. VCmax and ACmax can be considered as the most sensitive indicators of this cholinergic deficiency.

Decidualized and pre-decidualized normal endometrial stromal cells produce more O-linked N-acetylglucosamine containing epitope H than non-decidualized normal endometrial stromal cells.

The epitope H contains an O-linked N-acetylglucosamine residue in a specific conformation and/or environment recognized by the monoclonal antibody H (mAbH). mAbH stains two bands with Mr x10(-3) of 209 and 62 in lysates of cultured rat astrocytes. In addition, in extracts of cultured MCF-7 breast carcinoma cell line cells it stains cytokeratin 8 and five polypeptides originating from Triton X-100-soluble (Mr x10(-3) of 232, 67 and 37) and from the Triton X-100-insoluble (Mr x10(-3) of 51 and 50) fractions, respectively. In our previous studies we used the mAbH to investigate by immunostaining the expression of the epitope H in normal human brains, human brains with a variety of lesions, astrocytic tumors, infiltrating ductal breast carcinomas, fibroadenomas, and mitochondria-rich normal, metaplastic and neoplastic cells. In order to gain further insight into the expression patterns of the epitope H in human tissues we used the mAbH to investigate the immunohistochemical expression of the epitope H in normal human endometrium, including 30 cases of proliferative endometrium, 30 cases of early secretory endometrium, 30 cases of mid secretory endometrium, 30 cases of late secretory endometrium and 30 cases of decidual tissues. The main results were the following: 1) The decidual stromal cells presented in all cases high cytoplasmic expression of the epitope H; 2) The pre-decidual stromal cells presented in all cases of late secretory endometrium significant cytoplasmic expression of the epitope H ranging from moderate to high expression; 3) The non pre-decidual stromal cells of the functional endometrial layer presented in all cases insignificant cytoplasmic expression of the epitope H ranging from null to low expression; 4) The stromal cells of the basal layer of the endometrium and decidua did not express the epitope H in any case; 5) The endometrial stromal granulocytes did not express the epitope H in any case and 6) The blood vessel wall cells (endothelial and smooth muscle) of the endometrium through the whole duration of the menstrual cycle and of the decidua presented high cytoplasmic expression of the epitope H. It is concluded that decidualized and pre-decidualized human normal endometrial stromal cells show increased expression of the O-linked N-acetylglucosamine containing epitope H compared to non-decidualized endometrial stromal cells. These findings suggest that the expression of the epitope H may be under positive progesteronic control in normal human endometrium. Further investigation of the antigens bearing the epitope H might help to gain further insight into the histophysiology and the pathology of human endometrium.

Evaluation of the antimicrobial effectiveness of citric acid and sodium hypochlorite on the anaerobic flora of the infected root canal.

This study was designed to evaluate the antimicrobial effectiveness of 25% citric acid, compared with 2.5% sodium hypochlorite, after time intervals of 5, 15, 30 and 60 min. Anaerobic bacterial species were isolated from infected root canals and categorized according to Gram stain as Gram-positive cocci, Gram-negative cocci, Gram-positive rods and Gram-negative rods. The experimental method used was similar to that proposed by the German Society for Hygiene and Microbiology (DGHM) with slight modifications. Testing revealed that citric acid does demonstrate antimicrobial properties against anaerobic bacteria, especially against cocci, but it is less effective than 2.5% sodium hypochlorite.

In vitro evaluation of the effectiveness of calcium hydroxide and paramonochlorophenol on anaerobic bacteria from the root canal.

The effectiveness of calcium hydroxide and paramonochlorophenol (PMCP) was tested on anaerobic bacteria isolated from infected root canals, at time intervals 5, 15, 30 and 60 min. The experimental method used was similar to that proposed by the German Society of Hygiene and Microbiology (DGHM), with slight modifications. Testing revealed that calcium hydroxide was significantly more effective than PMCP against the total number of anaerobes. Additionally, it was proved to be quickly and highly effective against some microorganisms related to severe clinical symptoms, such as B. melaninogenicus and P. gingivalis, as well as against actinomyces species.

In vitro study of the indirect action of calcium hydroxide on the anaerobic flora of the root canal.

The aim of this study was to evaluate in vitro a possible mechanism involved in the antimicrobial action of calcium hydroxide, namely absorption of carbon dioxide from the root canal. Twenty obligate and 20 facultative anaerobic bacteria isolated from infected root canals and identified to species level were used. For each bacterial species a standard concentration was achieved and 0.1 ml of the inoculum was spread on blood agar plates, which were consecutively incubated in an anaerobic chamber for 5-7 days. One experimental and one control group were studied: the experimental group included one plate with the bacterial species as well as one open plate containing 32 g calcium hydroxide paste at a mixing ratio of 6:4. Both plates were incubated in an anaerobic chamber for 72 h. The control group included only one plate containing the same bacterial species and was incubated under the same conditions. After a 72-h incubation, the number of the recovered bacteria were counted in both groups. Statistical analysis showed that the number of bacteria recovered from the control group was significantly lower than that of the experimental group, but no particular resistance of any bacterial species to calcium hydroxide could be detected. This finding strongly suggests that the ability of calcium hydroxide to absorb carbon dioxide may contribute to its antibacterial activity.

Subgingival microflora in smokers with early onset periodontitis.

Cigarette smoking is a potent risk factor which has recently been associated with periodontal disease progression. The objective of this study was to detect the microbial profile of early onset periodontitis in smokers and compare it to that of non-smokers. The study population consisted of 50 systemically healthy individuals aged 25 to 38 years, exhibiting early onset periodontitis. 25 patients were smokers (> 20 cigarettes/day) and 25 non-smokers. Two pooled bacterial samples comprised of four periodontal sites with probing depth > 5 mm each, were collected from each individual. The samples were cultured aerobically and anaerobically for bacterial isolation using selective and non-selective media. Isolates were characterized to species level by conventional biochemical tests and various identification kits. The differences in bacterial counts using the Mann Whitney U test were statistically significant for Staphylococcus aureus, Campylobacter concisus, Eikenella corrodens, Escherichia coli, Bacteroides forsythus, Bacteroides gracilis, Campylobacter rectus, Porphyromonas gingivalis, Selenomonas sputigena and Candida albicans in smokers. Statistically significant differences for Peptostreptococcus micros, Actinomyces naeslundii, Eubacterium lentum and Capnocytophaga gingivalis were detected in non-smokers. The isolation of bacteria belonging to the exogenous flora like E. coli, C. albicans and S. aureus in smokers microflora underscores the importance of the host which is adversely affected by cigarette smoking.

Inflammatory infiltrate of chronic periradicular lesions: an immunohistochemical study.

AIM: To determine the cellular profile of human chronic periradicular lesions using immunohistochemical methods in order to study the differences in the cell infiltrate of periradicular granulomas and cysts. METHODOLOGY: The study population consisted of 45 individuals without any systemic disease. Biopsies were obtained during periradicular surgery. Paraffin-embedded sections were stained by the avidin-biotin complex method (ABC), whilst cryostat tissue sections were stained using the alkaline phosphatase antialkaline phosphatase assay (APAAP). These methods are highly valid and sensitive using a panel of specific monoclonal antibodies: CD4, CD8, CD3, CD10, HLADR, CD20, CD45RO, CD68 and CD57. The 45 specimens were characterized by the use of both techniques. RESULTS: The 45 specimens were histologically diagnosed as: 25 periradicular granulomas, 17 periradicular cysts and 3 scar tissues. No statistically significant differences were detected in the inflammatory infiltrate between periradicular granulomas and cysts. Observation of the sections showed that the majority of inflammatory cells consisted of T and B lymphocytes and macrophages. T and B lymphocytes were equally distributed in 60% of the cases. The T4/T8 ratio ranged approximately from 1 to 3 and greater, being consistent with inflammation of periradicular tissues. The final differentiation of B lymphocytes to plasma cells was also detected, whilst natural killer (NK) cells were found in only 10 cases (22%). Moreover, antigen presenting cells and T suppressor/cytotoxic cells were found to be associated with both pre-existing and newly formed epithelium. CONCLUSIONS: Periradicular granulomas and cysts represent two different stages in the development of chronic periradicular pathosis as a normal result of the process of immune reactions that cannot be inhibited.

Microflora in adult periodontitis.

The subgingival microflora of adult periodontitis was studied in 8 adults (36-47 years) and compared with that of 10 periodontally healthy individuals (36-43 years). A total of 64 periodontal lesions were examined, and classified according to the attachment level in three categories: attachment loss > 6 mm, attachment loss 4-6 mm and attachment loss < 4 mm. Also for comparative purposes 20 gingival sulci were evaluated. Samples were taken using three standardized paper points and were incubated anaerobically in selective and non-selective media. The results showed a statistically significant association of Capnocytophaga gingivalis and Capnocytophaga sputigena with moderate periodontal lesions, while Haemophilus segnis has been correlated to severe periodontal lesions. We concluded that C. gingivalis, C. sputigena and H. segnis might be potentially conductive to periodontal deterioration in adult periodontitis.

Predominant microflora of severe, moderate and minimal periodontal lesions in young adults with rapidly progressive periodontitis.

The purpose of this investigation was to study the microflora of severe, moderate and minimal periodontal lesions, in young adults with rapidly progressive periodontitis (RPP). Subgingival plaque samples were taken from 142 periodontal lesions in 10 young adults aging 25 to 35 years. The examination of the subgingival microflora indicated that certain species, including Porphyromonas gingivalis, Bacteroides forsythus, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, and Campylobacter species were found to be predominant in severe periodontal lesions. B. forsythus, P. gingivalis, Prevotella intermedia, F. nucleatum, Capnocytophaga ochracea, were predominant in medium lesions while Streptococcus species and Actinomyces species, C. ochracea, Haemophilus segnis and Veillonella parvula, were found in higher levels in minimal periodontal lesions.

The direct immunofluorescence in the presumptive identification of Bacteroides species.

Two kits containing fluorescein labeled-polyvalent antisera to Bacteroides fragilis, Bacteroides distasonis, Bacteroides ovatus, Bacteroides thetaiotaomicron, Bacteroides vulgatus (Fluoretec-F) and to Bacteroides melaninogenicus subsp. melaninogenicus and intermedius and to Bacteroides asaccharolyticus (non-oral strains) (Fluoretec-M) were evaluated for the identification of the respective organisms in bacterial cultures and clinical specimens. In both cases immunofluorescent tests were compared to the bacteriological data. The results clearly demonstrated that Fluoretec-F has a specificity 97% and a sensitivity 96%. The corresponding values for Fluoretec-M were 99% and 87.5% respectively, either clinical specimens or pure cultures were examined. The presence of aerobic of facultative anaerobic bacteria and obligate anaerobic bacteria other than those to which antisera were included in the test kits did not seem to influence the test result. No cross-reactivity between Fluoretec-F and -M were observed. Also, no positive fluorescence was obtained when other anaerobic bacteria than Bacteroides spp. were tested. The same was also the case with various aerobic or facultative anaerobic bacteria. The use of Fluoretec-F and -M is considered as a useful tool for the early accurate identification of the Bacteroides species mentioned, which may serve as a guide for an appropriate prompt therapy which is especially needed in serious anaerobic infections.