BRCA1 Immunohistochemical Staining as a Prognostic Indicator in Uterine Serous Carcinoma

OBJECTIVES: The objective of this study was to investigate the relationship between BRCA1 protein expression, as determined by immunohistochemistry, and clinical outcome in uterine serous carcinoma (USC). METHODS: A tissue microarray containing duplicate cores of 73 cases of USC was immunohistochemically stained with mouse anti-BRCA1 (Ab-1) mouse monoclonal (MS110) antibody. The cores were scored in a semiquantitative manner evaluating both the distribution and intensity of nuclear staining. BRCA1 protein expression was correlated with progression-free survival. RESULTS: Seventy-two of 73 cases were assessable, and there was a statistically significant decreased progression-free survival for those cases exhibiting tumor cell nuclei staining of 76% or greater (P = 0.0023). CONCLUSIONS: Our study illustrates that a low level of BRCA1 protein expression is a favorable prognostic indicator in USC, similar to what is observed in high-grade serous ovarian carcinoma. Further studies should focus on the BRCA1 status of USCs at a molecular level and also investigate whether BRCA1 protein expression is associated with response to chemotherapy in USC.

A rapid staining-assisted wood sampling method for PCR-based detection of pine wood nematode Bursaphelenchus xylophilus in Pinus massoniana wood tissue

For reasons of unequal distribution of more than one nematode species in wood, and limited availability of wood samples required for the PCR-based method for detecting pinewood nematodes in wood tissue of Pinus massoniana, a rapid staining-assisted wood sampling method aiding PCR-based detection of the pine wood nematode Bursaphelenchus xylophilus (Bx) in small wood samples of P. massoniana was developed in this study. This comprised a series of new techniques: sampling, mass estimations of nematodes using staining techniques, and lowest limit Bx nematode mass determination for PCR detection. The procedure was undertaken on three adjoining 5-mg wood cross-sections, of 0.5 · 0.5 · 0.015 cm dimension, that were cut from a wood sample of 0.5 · 0.5 · 0.5 cm initially, then the larger wood sample was stained by acid fuchsin, from which two 5-mg wood cross-sections (that adjoined the three 5-mg wood cross-sections, mentioned above) were cut. Nematode-staining-spots (NSSs) in each of the two stained sections were counted under a microscope at 100· magnification. If there were eight or more NSSs present, the adjoining three sections were used for PCR assays. The B. xylophilus – specific amplicon of 403 bp (DQ855275) was generated by PCR assay from 100.00% of 5-mg wood cross-sections that contained more than eight Bx NSSs by the PCR assay. The entire sampling procedure took only 10 min indicating that it is suitable for the fast estimation of nematode numbers in the wood of P. massonina as the prelimary sample selections for other more expensive Bx-detection methods such as PCR assay.

COLOR STABILITY of SEALED COMPOSITE RESIN RESTORATIVE MATERIALS AFTER ULTRAVIOLET ARTIFICIAL AGING and IMMERSION IN STAINING SOLUTIONS

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of Different Light-Curing Modes on Degree of Conversion, Staining Susceptibility and Stain's Retention Using Different Beverages in a Nanofilled Composite Resin

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparison between histochemical and immunohistochemical methods for diagnosis of sporotrichosis

Aims: To compare the efficacy of histochemical and immunohistochemical methods in detecting forms of Sporothrix schenckii in tissue. Methods: Thirty five cutaneous biopsy specimens from 27 patients with sporotrichosis were stained by histochemical haemotoxylin and eosin, periodic acid Schiff, and Gomori's methenamine silver methods and an immunohistochemical (avidin-biotin complex immunoperoxidase) (ABC) technique associated with a newly produced rabbit polyclonal antibody anti-Sporothrix schenckii. Results: A total of 29 (83%) cases were positive by the ABC method used in association with anti-Sporothrix schenckii rabbit polyclonal antibodies. Histochemical methods, using silver staining, periodic acid Schiff, and conventional haematoxylin and eosin detected 37%, 23%, and 23% of forms of S schenckii, respectively. The ABC technique was significantly more reliable than periodic acid Schiff and silver staining techniques. Conclusions: It is concluded that immunostaining is an easy and rapid method which can efficiently increase the accuracy of the diagnosis of sporotrichosis in human tissue.

Staining of esthetic brackets by plaque disclosing solutions

AIM: To evaluate the staining of esthetic orthodontic brackets by plaque disclosing solutions. METHODS: Two types of brackets manufactured by GAC/DENTSPLY(r) were evaluated: ceramic (n=30) and polycarbonate (n=30). The brackets were divided into 6 groups. Two control groups (n=6) were immersed in absolute ethanol: GI - ceramic brackets and GII - polycarbonate brackets. Four experimental groups (n=12) were immersed in different plaque disclosing solutions: GIII (ceramic brackets) and GIV (polycarbonate brackets) were immersed in Replak(r); GV (ceramic brackets) and GVI (polycarbonate brackets) were immersed in Replasul "S"(r). Relative quantitative analysis of the influence of plaque disclosing tablets on bracket staining was performed using reflectance spectrophotometry of stain deposition. Exploratory analysis of the data was performed using Analysis of Variance (ANOVA) in a 2x2 factorial setup (bracket x immersion) with additional treatments (controls). RESULTS: The results demonstrated that the ceramic brackets presented the highest amount of staining when Replasul "S"(r) was used (pd"0.05). However, when Replak(r) was used, no statistically significant difference was found in comparison with the control group (p>0.05). For polycarbonate brackets, staining was detected for both disclosing solutions (p>0.05). CONCLUSIONS: The disclosing solutions caused stain formation on polycarbonate brackets and, under the tested conditions, use of Replak(r) on ceramic brackets did not cause staining.

Color Stability of Resin Used for Caries Infiltration After Exposure to Different Staining Solutions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Lectinhistochemical staining of granuloma induced by bacillus Calmette- Guerin in Piaractus mesopotamicus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of different periods of preconditioning with saliva on adhesion of C. albicans to a denture base resin by crystal violet staining and XTT assay.

Aim: The role of saliva on Candida adhesion to biomaterials has not been clearly defined. The present study investigates whether different periods of preconditioning with saliva would influence the adhesion of Candida albicans to a denture base resin. Methods: Ninety samples of acrylic resin with smooth surfaces were made and then divided into five groups: one control without saliva, and four experimental groups conditioned in saliva for periods of 30 min, 1, 3, or 12 h. Candida adhesion was evaluated by crystal violet staining and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-([phenylamino] carbonyl)-2H-tetrazolium-hydroxide assay. Results: The one-way analysis of variance revealed that there were no significant differences among the mean number of adherent cells or among the mean absorbance for all groups. No significant correlation was found between the two methods used for assessing Candida albicans adhesion. Conclusion: The different periods of preconditioning with saliva had no significant influence on the adhesion of Candida albicans to the denture base acrylic resin.

Effect of staining agents on color change of composites

Composite resins are materials that can present color changing when exposed to pigments. Objective: The aim of this study was to evaluate, in vitro, the color changing of composites after immersion in different substances for different periods. Material and methods: Two microhybrid composite resins: Charisma (Heraeus – Kulzer) and Opallis (FGM) were used. Red wine and acai pulp were also used as immersion medium. For this study, 32 specimens with 10 mm of diameter and 2 mm of thickness were used, divided into 4 groups: Group 1 – Opallis composite immersed in red wine solution; Group 2 – Opallis composite immersed in acai berry pulp solution; Group 3 – Charisma composite immersed in red wine solution; Group 4 – Charisma composite immersed in acai berry pulp solution. The specimens were evaluated in the following time periods: T0 – baseline, T1 – 24 hours, T2 – 48 hours, T3 – 72 hours and T4 – 96 hours. For the assessment of staining, a spectrophotometer for colorimetry was used (Color Guide 45 / 0, PCB 6807 BYK-Gardner Gerestsried GmBH, Germany), and the values obtained were transferred to a computer and recorded according to CIELAB system. Results: The data were evaluated using Kruskal- Wallis non-parametric tests with the following mean values for the immersion periods of 24, 48, 72 and 96 hours, respectively: G1 – 7.35, 7.84, 9.04,10.48; G2 – 2.92, 4.15, 4.30, 4.64; G3 – 3.14, 7.35, 8.13, 8.43, G4 – 4.49, 5.99, 6.92, 6.76. Conclusion: Red wine showed a higher tendency toward altering the composite color than acai berry pulp. In addition, no significant difference was found concerning to the behavior of the two composite resins. Concerning to the immersion time periods, significant differences were only observed among the groups in the 24 hour time period.

Taste alteration, mouth dryness and teeth staining as side effects of medications taken by elderly

Elderly patients generally use several types of medication, some of which may cause oral side effects. Aim: To investigate the oral side effects caused by medication in an elderly sample. Methods: Three hundred patients were interviewed about their use of medication and were divided in two groups: institutionalized (n=150) and community-dwelling (n=150) elderly. Results: The most used drugs were antihypertensives (53%) for community-dwelling elders and antiulceratives (76%) for the institutionalized ones. The more prevalent side effects were taste alterations that occurred in 19%, dry mouth in 17% and teeth staining in 2%. Conclusions: A high prevalence of oral side effects from medications used by the elderly was found in this study. The health professionals should be aware of the possible side effects caused by prescribed medications.

Fluorescence Intensity of Composite Layering Combined with Surface Sealant Submitted to Staining Solutions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Staining of the Internal Limiting Membrane with the Use of Heavy Brilliant Blue G

Background: Brilliant blue G (BBG) is frequently used in chromovitrectomy to facilitate internal limiting membrane (ILM) peeling. A study was initiated to evaluate if heavy BBG is safe and effective in staining the ILM. Methods: We studied 30 eyes, 23 with idiopathic macular holes and 7 of patients with diabetic macular edema. Removal of the ILMs was assisted by heavy BBG staining. In cases with histopathological correlation the ILMs were evaluated with hematoxylin and eosin, Masson's trichrome, periodic acid-Schiff and glial fibrillary acidic protein staining. In addition, immunohistochemistry was also performed using specific antibodies for vimentin, neuron-specific enolase, factor VIII and CD68. Using the Image-Pro Plus software of Media Cybernetics Co. we found an average thickness in ILMs. Results: Of the ILM specimens sent, 19/30(63.33%) could not be processed properly because of the limited sample material, recognizing only fragments of dispersed fibrillar material. In macular hole ILMs we found an average thickness of 1.3 +/- 0.65 mu m, and in diabetic macular edema ILMs an average thickness of 6.2 +/- 1.4 mu m. Conclusions: In heavy BBG-assisted ILM peeling we observed no intraoperative or postoperative complications after a mean follow-up of 12 months. Heavy BBG could be an effective and safe vehicle for staining the ILM. Copyright (C) 2012 S. Karger AG, Basel

Comparison of two laboratory-developed PCR methods for the diagnosis of Pulmonary Tuberculosis in Brazilian patients with and without HIV infection

Abstract Background Direct smear examination with Ziehl-Neelsen (ZN) staining for the diagnosis of pulmonary tuberculosis (PTB) is cheap and easy to use, but its low sensitivity is a major drawback, particularly in HIV seropositive patients. As such, new tools for laboratory diagnosis are urgently needed to improve the case detection rate, especially in regions with a high prevalence of TB and HIV. Objective To evaluate the performance of two in house PCR (Polymerase Chain Reaction): PCR dot-blot methodology (PCR dot-blot) and PCR agarose gel electrophoresis (PCR-AG) for the diagnosis of Pulmonary Tuberculosis (PTB) in HIV seropositive and HIV seronegative patients. Methods A prospective study was conducted (from May 2003 to May 2004) in a TB/HIV reference hospital. Sputum specimens from 277 PTB suspects were tested by Acid Fast Bacilli (AFB) smear, Culture and in house PCR assays (PCR dot-blot and PCR-AG) and their performances evaluated. Positive cultures combined with the definition of clinical pulmonary TB were employed as the gold standard. Results The overall prevalence of PTB was 46% (128/277); in HIV+, prevalence was 54.0% (40/74). The sensitivity and specificity of PCR dot-blot were 74% (CI 95%; 66.1%-81.2%) and 85% (CI 95%; 78.8%-90.3%); and of PCR-AG were 43% (CI 95%; 34.5%-51.6%) and 76% (CI 95%; 69.2%-82.8%), respectively. For HIV seropositive and HIV seronegative samples, sensitivities of PCR dot-blot (72% vs 75%; p = 0.46) and PCR-AG (42% vs 43%; p = 0.54) were similar. Among HIV seronegative patients and PTB suspects, ROC analysis presented the following values for the AFB smear (0.837), Culture (0.926), PCR dot-blot (0.801) and PCR-AG (0.599). In HIV seropositive patients, these area values were (0.713), (0.900), (0.789) and (0.595), respectively. Conclusion Results of this study demonstrate that the in house PCR dot blot may be an improvement for ruling out PTB diagnosis in PTB suspects assisted at hospitals with a high prevalence of TB/HIV.

A randomized, controlled clinical trial on the clinical, microbiological, and staining effects of a novel 0.05% chlorhexidine/herbal extract and a 0.1% chlorhexidine mouthrinse adjunct to periodontal surgery

BACKGROUND: Chlorhexidine (CHX) rinsing after periodontal surgery is common. We assessed the clinical and microbiological effects of two CHX concentrations following periodontal surgery. MATERIALS AND METHODS: In a randomized, controlled clinical trial, 45 subjects were assigned to 4 weeks rinsing with a 0.05 CHX/herbal extract combination (test) or a 0.1% CHX solution. Clinical and staining effects were studied. Subgingival bacteria were assessed using the DNA-DNA checkerboard. Statistics included parametric and non-parametric tests (p<0001 to declare significance at 80% power). RESULTS: At weeks 4 and 12, more staining was found in the control group (p<0.05 and p<0.001, respectively). A higher risk for staining was found in the control group (crude OR: 2.3:1, 95% CI: 1.3 to 4.4, p<0.01). The absolute staining reduction in the test group was 21.1% (9 5% CI: 9.4-32.8%). Probing pocket depth (PPD) decreases were significant (p<0.001) in both groups and similar (p=0.92). No rinse group differences in changes of bacterial counts for any species were found between baseline and week 12. CONCLUSIONS: The test CHX rinse resulted in less tooth staining. At the study endpoint, similar and high counts of periodontal pathogens were found.