Actinomyces denticolens colonisation identified in equine tonsillar crypts.

Recently, submandibular abscesses associated with Actinomyces denticolens have been reported in horses. The actinomycotic clumps have been observed in the tonsillar crypts. The aim of this study was to demonstrate colonisation of A denticolens in equine tonsils. Twelve equine tonsils obtained from a slaughterhouse were divided into two parts for histopathological examination and for isolation of A denticolens. When actinomycotic clumps were found in these tonsillar crypts, immunohistochemistry using hyperimmune serum against A denticolens (DMS 20671) was performed on the serial sections. To determine whether Actinomyces-like bacteria isolated using immunoantigenic separation technique were A denticolens, the isolates were analysed for the 16S rRNA gene sequence. Actinomycotic clumps were found in the tonsillar crypts of 11 (91.7 per cent) horses. The clumps were of the saprophytic type accompanied with the feedstuffs, but a few clumps were surrounded by inflammatory cells. A denticolens antigens were immunodetected not only in the clumps of 11 (100 per cent) tonsils, but also in the tonsillar parenchyma. Six isolates obtained from four tonsils showed 99.7-99.9 per cent similarity to A denticolens in the 16S rRNA gene sequence. In horses, the colonisation sites of A denticolens are the tonsils, thus the authors suggest that the tonsils provide the intrinsic infection site for A denticolens.

Analysis of circulating human and viral microRNAs in patients with congenital cytomegalovirus infection.

OBJECTIVE: Cytomegalovirus (CMV) is the most common cause of congenital infection and can cause neurodevelopmental disabilities, although a majority of patients are asymptomatic. Biomarkers associated with disease severity would be desirable to distinguish asymptomatic from mildly symptomatic patients who may benefit from antiviral treatment. MicroRNAs (miRNAs) are noncoding RNAs that may have the potential to serve as biomarkers. STUDY DESIGN: Thirteen infants with congenital CMV infection were enrolled, and plasma levels of 11 human- and 3 CMV-encoded miRNAs were quantitated by real-time PCR. Plasma levels of miRNAs and their associations with clinical features were evaluated. RESULTS: The levels of miR-183-5p and miR-210-3p were significantly higher in patients with congenital CMV infection than in control infants, whereas no significant associations between levels of miRNAs and clinical features of congenital CMV infection were observed. CONCLUSION: Plasma miRNAs could be associated with the pathogenesis of congenital CMV infection and could be used as disease biomarkers.

The problems of cervical conization for postmenopausal patients.

PURPOSE: Cervical conization is the definitive treatment for women of any age who have cervical intraepithelial neoplasia (CIN). However, complications of the procedure have not been fully investigated in postmenopausal patients. The aim of this retrospective study was to evaluate the results and complications of cervical conization performed on premenopausal and postmenopausal patients. MATERIALS AND METHODS: This study recruited 405 patients who had undergone cervical laser conization. The median age was 36 years (range 20 to 75), and there were 361 (89.1%) premenopausal and 44 (10.9%) postmenopausal women. RESULTS: The length of the cone removed from the postmenopausal patients was significantly longer than the length from the premenopausal patients (17.9 ± 3.9 mm vs. 15.7 ± 3.6 mm, respectively; p = 0.02). The rate of positive endocervical cone margins from the premenopausal patients was significantly higher than the rate from the postmenopausal patients (9.1% vs. 0%, respectively; p = 0.037). The rate of cervical stenosis was significantly higher in postmenopausal patients than in premenopausal patients (59.1% vs. 8.3%; respectively; p < 0.0001). There was no difference in the rates of frequency of intraoperative complications. CONCLUSIONS: Although deep incision is mandatory for complete excision of CIN in postmenopausal patients, it increases the incidence of cervical stenosis. Cervical conization may be a less invasive surgical procedure for older women with CIN than hysterectomy; however, the risk of postoperative complications remains, causing a dilemma for physicians treating postmenopausal women with CIN.

Effectiveness of a simulated patient training programme based on trainee response accuracy and appropriateness of feedback.

INTRODUCTION: Simulated patients (SPs) need education and training in required skills to be effective resources in education. This study was conducted to examine the effectiveness of an SP training programme based on the accuracy of trainee responses and the appropriateness of their feedback. METHODS: Thirty-two applicants to the training programme and 35 experienced SPs were included in this study. The experienced SPs served as a reference group. The rate of accurate responses and the rate of appropriate feedback were assessed with pre- and post-training tests, and these two outcome measures were compared with those of the experienced SPs. RESULTS: No significant differences were found in trainee response accuracy or appropriateness of feedback between pre- and post-training tests. The response accuracy rate of the trainees on the pre-training test was significantly lower than that of SPs with 1-2 years of experience, whilst there was no significant difference between these SPs and the trainees on the post-training test. CONCLUSIONS: Although our study suggests that more training is needed to improve the skills of SPs, the training programme may contribute to helping trainees reach a novice level in the skill of providing accurate responses. SP training should be encouraged to contribute to the effectiveness of such teaching and to establish the validity of the assessment.

Anti-interferon-α neutralizing antibody is associated with nonresponse to pegylated interferon-α plus ribavirin in chronic hepatitis C.

Pegylated interferon-α (PEG-IFN-α) plus ribavirin (RBV) treatment fails to achieve a sustained virological response (SVR) in approximately 20-50% of patients with chronic hepatitis C virus (HCV) infection. We assessed the contribution of an anti-IFN-α neutralizing antibody (NAb) on the nonresponse to treatment. NAbs were detected using an antiviral assay that assessed the neutralizing effects of serum samples against IFN. Serum samples were obtained at the end of the treatment and evaluated for the presence of NAbs using recombinant IFN-α as a standard. We studied 129 PEG-IFN-α/RBV-treated patients. In the 82 end-of-treatment responders, no NAbs were detected. Of the 47 patients who did not respond, seven (15%) were positive for NAbs. We also examined an additional 83 patients who had not responded to PEG-IFN-α treatment, and detected 12 with NAbs. Patients with good IFN-responsive characteristics, including HCV genotype 2/3 and major allele homozygotes for interleukin-28B, were included in the 19 patients with NAbs. No NAbs interfered with the antiviral activity of natural human IFN-ß (nIFN-ß) and re-treatement of patients with NAbs with nIFN-ß/RBV achieved SVR. Our analyses revealed that the emergence of anti-IFN-α NAbs was a candidate causal factor of PEG-IFN-α-treatment failure. Therefore, these antibodies should be assayed in patients who do not respond to PEG-IFN-α therapy, and if detected, other effective treatments, i.e., medications that are not neutralized by anti-IFN-α NAbs, should be considered.

Self-assembled Nano-layering at the Adhesive interface.

According to the 'Adhesion-Decalcification' concept, specific functional monomers within dental adhesives can ionically interact with hydroxyapatite (HAp). Such ionic bonding has been demonstrated for 10-methacryloyloxydecyl dihydrogen phosphate (MDP) to manifest in the form of self-assembled 'nano-layering'. However, it remained to be explored if such nano-layering also occurs on tooth tissue when commercial MDP-containing adhesives (Clearfil SE Bond, Kuraray; Scotchbond Universal, 3M ESPE) were applied following common clinical application protocols. We therefore characterized adhesive-dentin interfaces chemically, using x-ray diffraction (XRD) and energy-dispersive x-ray spectroscopy (EDS), and ultrastructurally, using (scanning) transmission electron microscopy (TEM/STEM). Both adhesives revealed nano-layering at the adhesive interface, not only within the hybrid layer but also, particularly for Clearfil SE Bond (Kuraray), extending into the adhesive layer. Since such self-assembled nano-layering of two 10-MDP molecules, joined by stable MDP-Ca salt formation, must make the adhesive interface more resistant to biodegradation, it may well explain the documented favorable clinical longevity of bonds produced by 10-MDP-based adhesives.

Predicting nedaplatin sensitivity of cervical cancer using the histoculture drug response assay.

PURPOSE: There are currently no clinically available chemosensitivity assays for cervical cancer. In this study we evaluated whether the histoculture drug response assay (HDRA) could be used to predict chemosensitivity to nedaplatin (NDP) in cervical cancer. METHODS: Fifty-four surgical specimens and biopsies from patients with squamous cell carcinoma of the cervix were tested with the HDRA. The results were used to calculate the concentration resulting in 50% inhibition of tumor growth (IC50). We then determined the cut-off concentration for NDP, and investigated the chemosensitivity of NDP for each patient. Moreover, the correlations between chemosensitivity and the clinical response of NDP-containing chemotherapy, and the clinical outcomes of the patients with Stage I and II disease were also investigated. RESULTS: Fifty-one of 54 specimens (94.0%) were evaluable with this assay. The optimal cutoff concentration of NDP was determined to be 48 microg/ml. In 18 patients with measurable lesions, all nine patients in the high sensitive group by HDRA were judged as partial response (PR) to NDP containing chemotherapy. In contrast five of nine patients in the low sensitive group were classified as stable disease, and four were PR. The true positive rate was 100%, the true negative rate was 55.6%, and the accurate prediction rate was 77.8%. Furthermore, the disease-free survival of the high sensitive group tended to be better than that of the low sensitive group in the patients who received postoperative adjuvant chemotherapy with NDP. CONCLUSIONS: In the current study, the sensitivity of cervical tumors to nedaplatin was predicted by the HDRA.

Self-etch monomer-calcium salt deposition on dentin.

Previous research, in which the bonding effectiveness of the 3 self-etch monomers HAEPA, EAEPA, and MAEPA was determined, showed that MAEPA was most effective. In this study, the molecular interactions of these monomers with hydroxyapatite and dentin were investigated by combining x-ray diffraction, infrared spectroscopy, and scanning electron microscopy. We tested the null hypothesis that the bonding performance of these monomers does not correlate to the formation of monomer-calcium salts and to hydrolytic stability of these monomer-calcium complexes. Monomer/ethanol/water solutions were prepared and applied to synthetic hydroxyapatite and dentin. While HAEPA and EAEPA dissolved dentin considerably and deposited unstable calcium-phosphate salts (DCPD), MAEPA formed hydrolysis-resistant monomer-calcium salts that remained attached to the dentin surface even after being washed. The chemical stability of the monomer-Ca salts was thought to contribute in particular to the bond durability, but this study shows that the formation of stable monomer-calcium salts also enhances the 'immediate' bonding performance of self-etch adhesives.

Ultrasound predictors of mortality in monochorionic twins with selective intrauterine growth restriction.

OBJECTIVES: The aim of this study was to evaluate the use of ultrasound assessment to predict risk of mortality in expectantly managed monochorionic twin fetuses with selective intrauterine growth restriction (sIUGR). METHODS: This was a retrospective study of 101 monochorionic twin pregnancies diagnosed with sIUGR before 26 weeks of gestation. All patients were under expectant management during the observation period. At the initial evaluation, the presence or absence of each of the following abnormalities was documented: oligohydramnios; stuck twin phenomenon; severe IUGR < 3(rd) centile of estimated fetal weight; abnormal Doppler in the umbilical artery; and polyhydramnios in the larger twin. The relationships between these ultrasound findings and mortality of sIUGR fetuses were evaluated using multiple logistic regression analysis. RESULTS: Of 101 sIUGR twins, 22 (21.8%) fetuses suffered intrauterine demise and nine (8.9%) suffered neonatal death; 70 (69.3%) survived the neonatal period. Multiple logistic regression analysis revealed that the stuck twin phenomenon (odds ratio (OR): 14.5; 95% CI: 2.2-93.2; P = 0.006) and constantly absent diastolic flow in the umbilical artery (OR: 29.4; 95% CI: 3.3-264.0; P = 0.003) were significant risk factors for mortality. CONCLUSIONS: Not only abnormal Doppler flow in the umbilical artery but also severe oligohydramnios should be recognized as important indicators for mortality in monochorionic twins with sIUGR.

Monochorionic monoamniotic twin pregnancies with two yolk sacs may not be a rare finding: a report of two cases.

The exact determination of amnionicity is a major issue for the clinical management of monochorionic twin pregnancies, due to the high risk of perinatal mortality and morbidity in monochorionic monoamniotic (MCMA) twins. Counting the number of yolk sacs is believed to be a good indicator of amnionicity in the early first trimester, and it has previously been suggested that the number of yolk sacs is equal to amnionicity in both MCMA and monochorionic diamniotic twin pregnancies. However, the accuracy of the relationship between number of yolk sacs and amnionicity has recently been called into question. To the best of our knowledge, no previous reports have shown two yolk sacs in MCMA twin pregnancies. We report two cases of MCMA twins with two yolk sacs on first-trimester ultrasonography, and confirmed monoamnionicity in the second trimester showing umbilical cord entanglement. Postnatal examination showed an MCMA placenta in both cases, and entangled umbilical cords confirmed monoamnionicity. The possibility of monoamnionicity must still be suspected when two yolk sacs are detected early in the first trimester on ultrasound examination in monochorionic twin pregnancies.

Validation of Quintero stage III sub-classification for twin-twin transfusion syndrome based on visibility of donor bladder: characteristic differences in pathophysiology and prognosis.

OBJECTIVE: To validate the Quintero stage III subclassification for twin-twin transfusion syndrome (TTTS) based on visibility of the bladder of the donor twin. METHODS: Between July 2002 and August 2006, there were 131 pregnant Japanese women affected by severe TTTS before 26 weeks' gestation, treated with fetoscopic laser surgery at five centers in Japan, whose pregnancies continued beyond 22 weeks. Outcome data were available in all cases and surviving infants were followed up for at least 6 years. This study focused on the Stage III TTTS patients. These were subclassified into Stage III atypical (abnormal Doppler flow with visible donor bladder) and Stage III classical (abnormal Doppler flow with non-visible donor bladder) groups. Perioperative data and postnatal outcomes were compared between the groups. RESULTS: Seven Stage I, 22 Stage II, 82 Stage III and 20 Stage IV pregnancies continued beyond 22 weeks. There was a significantly higher incidence of absent or reversed end-diastolic velocity in the umbilical artery (UA-AREDV) of the donor in Stage III atypical than in Stage III classical patients (83.8% vs. 53.3%, P = 0.004). Stage III atypical cases also had a significantly higher incidence of arterioarterial (AA) anastomoses (72.9% vs. 17.8%, P < 0.001) and intrauterine fetal demise (IUFD) of the donor (43.2% vs. 13.3%, P = 0.002). However, there were no differences in overall survival or in abnormal brain scans of surviving infants. Donors with both UA-AREDV and AA anastomoses had a significantly higher incidence of IUFD compared with the others (53.3%, P < 0.001). CONCLUSIONS: Quintero stage III atypical was characterized by a high incidence of AA anastomoses and UA-AREDV of the donor, resulting in IUFD. Subclassification of Stage III based on visibility of the bladder of the donor twin was adequate for and compatible with differentiating prognosis and pathophysiology.

Cytological criteria of endometrial lesions with emphasis on stromal and epithelial cell clusters: result of 8 years of experience with intrauterine sampling.

OBJECTIVE: There are a number of unresolved issues in endometrial cytology. They include the significance of nuclear atypia for the diagnosis of grade1 adenocarcinoma (G1AC) and atypical endometrial hyperplasia (AEH), cytological criteria of endometrial hyperplasia without atypia, and recognition of stromal cell cluster (SC) and its distinction from epithelial cell cluster (EC). METHODS: We examined nuclear atypia, SC and EC in typical cases of five categories: normal endometrium (NEM), simple endometrial hyperplasia without atypia (SEH), complex endometrial hyperplasia without atypia (CEH), G1AC and grade2 adenocarcinoma (G2AC). We classified EC into four types: simple EC (SPEC), large regular EC (LREC), large irregular EC (LIEC) and small irregular EC (SIEC). Based on the results, we developed criteria of endometrial cytology and have evaluated 13 639 cases over 8 years. RESULTS: Nuclear atypia was significantly more frequent in G2AC than in any of the other four categories (P < 0.001). SC was significantly more frequent in NEM and SEH than in the other three categories (P < 0.001). G1AC and G2AC showed significantly higher frequency of LIEC than the other three categories (P < 0.001). CEH exhibited significantly higher frequency of LREC than the four categories (P < 0.001). The sensitivity and the specificity was 88.8% and 99.0% respectively. CONCLUSIONS: We could diagnose G1AC, G2AC and CEH with high accuracy using the established criteria mainly based on SC and EC. We think that the criteria may facilitate an effective screening and an objective interpretation of endometrial samples.

Sonography in total hip arthroplasty.

PURPOSE: The distance between the anterior surface of the neck of the prosthetic stem and the anterior joint capsule, the "capsular distance", is increased in total hip arthroplasty (THA) with synovitis. We evaluated the potential of ultrasonography (US) in measuring the "capsular distance" in THA hips one year after insertion. MATERIALS AND METHODS: We compared the measurements of the capsular distance using a ruler with those performed with US. A plastic pelvis and femur model with a prosthetic hip and paper tape to simulate the joint capsule were used. We also evaluated the intra- and interobserver agreements between 3 examiners of the US measurements of the anterior capsular distance in 22 patients with THA. The effect of experience in such type of examination was estimated. RESULTS: There was a high correlation when measuring the anterior capsular distance in the prosthetic hip model with a ruler as compared with US. The interobserver agreement in the US measurements was good and became better after examiners gained experience in this procedure. The intraobserver agreement was always better than the interobserver agreement and also improved with increasing numbers of examinations. CONCLUSION: Ultrasonography is a reliable method to measure the anterior capsular distance in THA, especially if performed by an experienced examiner.

Effect of 2-hydroxyethyl methacrylate pre-treatment on micro-tensile bond strength of resin composite to demineralized dentin.

The purpose of this study was to evaluate the effect of 2-hydroxyethyl methacrylate (HEMA) application on the micro-tensile bond strength of resin composite to demineralized dentin. Artificially demineralized lesions were formed on bovine dentin surfaces and treated with 10, 30, 50, 70 and 100 wt% HEMA aqueous solution. The surfaces were then applied and covered with SE Bond and AP-X according to the manufacturer's instruction. After immersion in 37 degrees C water for 24 h, bond strength were measured using a universal testing machine. Bond strengths to both demineralized dentin and normal dentin, without HEMA application, were also measured. Scanning electron microscopic (SEM) observation and confocal laser scanning microscopy (CLSM) analysis at the resin-dentin interface were also performed. The bond strength data were statistically compared with anova and Scheffe's test (P < 0.05). Bond strength to demineralized dentin treated with over 30 wt% HEMA aqueous solution were significantly higher than that to demineralized dentin without HEMA application, but significantly lower than that to normal dentin. SEM observation revealed that the hybrid layer and resin-tags thickened and lengthened with HEMA application. In CLSM, the diffusion of adhesive primer into demineralized dentin increased with HEMA application. These results indicated that HEMA application might increase the bond strength to demineralized dentin by the enhancement of resin monomer penetration of HEMA.

Micro-tensile bond strength of self-etching primer adhesive systems to human coronal carious dentin.

The aim of this study was to evaluate the micro-tensile bond strengths of three self-etching primer adhesive systems to normal dentin (ND), caries-affected dentin (CAD) and caries-infected dentin (CID). Human extracted molars with caries were used, and flat dentin surfaces ground by 600-grit SiC paper were prepared. The surfaces were dyed using Caries-Detector solution, treated with Clearfil SE Bond, Mac-Bond II and UniFil Bond, and then covered with resin composites according to manufacturer's instructions. After immersion in 37 degrees C water for 24 h, the teeth were serially sectioned into multiple slices. Each slice was distinguished into ND, CAD and CID groups by the degree of staining, and the bond strength was measured in a universal testing machine. Scanning electron microscopic (SEM) observation was also performed. For statistical analysis, anova and Scheffe's test were used (P < 0.05). The bond strengths of the three adhesive systems to CAD and CID were significantly lower than those to ND. There was significant difference in the bond strength to ND between Clearfil SE Bond and UniFil Bond, but no significant differences to CAD and CID among the three adhesive systems. On SEM, the hybrid layers in CAD and CID showed more porous structures compared with ND. The results indicated that the bond strengths to CAD and CID were not affected by a variety of self-etching primer adhesive systems because of the porous hybrid layer formation in carious dentin.

Current clinical results of the Tsukuba BNCT trial.

Nine high grade gliomas (5 glioblastomas and 4 anaplastic astrocytomas) were treated with BSH-based intaoperative boron neutron capture therapy (IOBNCT). BSH (100 mg/kg body weight) was intravenously injected, followed by single fraction irradiation using the mixed thermal/epithermal beam of Japan Research Reactor 4. The blood boron level at the time of irradiation averaged 29.9 (18.8-39.5)microg/g. The peak thermal neutron flux as determined by post-irradiation measurements varied from 1.99 to 2.77x10(9) n cm(-2)s(-1). No serious BSH-related toxicity was observed in this series. The interim survival data in this study showed median survival times of 23.2 months for glioblastoma and 25.9 months for anaplastic astrocytoma, results which are consistent with the current conventional radiotherapy with/without boost radiation. Of the 4 residual tumors, 2 showed complete response (CR) and 2 showed partial response (PR) within 6 months following BNCT. No linear correlation was proved between the dose and the occurrence of early neurological events. The maximum boron dose of 11.7-12.2 Gy in the brain related to the occurrence of radiation necrosis. The clinical application of a mixed thermal/epithermal beam and JRR-4 facilities on BSH-based IOBNCT proved to be safe and effective in this series.

Curved intertrochanteric varus osteotomy for osteonecrosis of the femoral head.

We reviewed the outcome of curved intertrochanteric varus osteotomy in the treatment of osteonecrosis of the femoral head in 20 hips. A mean varus angulation of 31 degrees was obtained by the osteotomy. The ratio of intact area on the weight-bearing portion increased from 19% to 61%. The mean elevation and lateral displacement of the greater trochanter were 1.2 cm and 0.5 cm, respectively. These changes in the position of the greater trochanter were very small when compared with those after conventional varus wedge osteotomy. Nonunion or delayed union was not observed. Quantitative analyses showed aggressive bone remodelling in the medial intertrochanteric region. Eighteen hips survived without collapse after a mean follow-up of 48 months. We conclude that curved varus osteotomy can be used to preserve the hip joint in patients with osteonecrosis of the femoral head.

Confocal laser scanning microscopic analysis of early plaque formed on resin composite and human enamel.

The purpose of this study was to analyse quantitatively the early bacterial plaque formed on resin composite and human enamel in vivo, using a confocal laser scanning microscope. Test pieces of resin composite and human enamel were retained at the buccal surfaces of the upper first molars of three volunteers for 4, 8 and 24 h to allow plaque formation. Then, the specimens were immersed in propidium iodide in phosphate-buffered saline to stain adherent bacteria and observed with a confocal laser scanning microscope. The ratios of the area occupied by microorganisms to the whole area of the optical field were calculated using a photo-image analysis system. The thickness of the plaque was also measured. Quantitative analysis revealed that the resin composite showed significantly higher bacterial adherence than human enamel throughout the test period. A difference was noticed in the morphology of the bacteria between the two groups. Our findings suggest that resin composite shows higher bacteria adherence during early plaque formation compared with human enamel. In addition, the present findings may suggest a presence of the difference in bacterial composition of plaque in both specimens.

Effect of fluoride-releasing adhesive system on decalcified dentin.

The aim of this study was to evaluate the effect of fluoride-releasing adhesive systems on human decalcified dentin in vitro. Two fluoride-releasing adhesive systems, Reactmer bond (RB, Shofu) and ABF (AF, Kuraray), an experimental system, and a commercial adhesive system without fluoride release, SE bond (SE, Kuraray), were used in this study. The amount of fluoride release from adhesive in deionized water was measured every week for 10 weeks. Class V cavities were prepared on extracted human pre-molars and decalcified dentin was promoted by using a bacterial caries induction system at the cavity floor. The cavities preserving decalcified dentin were restored with resin composite (AP-X, Kuraray) after treatment by each adhesive system. The specimens without treatment by adhesive system and restoration were used for control. The specimens with restoration were then incubated for 4 weeks at 37 degrees C, 100% humidity. Microradiographs of the specimens showed that the radiopacities of the decalcified dentin layers in RB and AF groups with fluoride release were significantly higher than those in SE or control groups without fluoride release. This result suggested that the fluoride-releasing adhesive systems enhanced mineralization of decalcified dentin.