The first case of chronic pulmonary Mycobacterium shigaense infection in an immunocompetent patient.

Mycobacterium shigaense has recently been recognized as an emerging human pathogen, and is well known as a skin pathogen in immunocompromised individuals. In this report we describe the first case of chronic pulmonary infectious disease caused by M. shigaense in an immunocompetent individual.

In tube immunocytochemistry for fluorescence-activated cell sorting that prevents RNA degradation in sorted cells.

Fluorescence-activated cell sorting (FACS) is a powerful tool for analyzing stem cells. When using fixed cells, however, it is sometimes difficult to analyze RNA extracted from sorted cells due to RNA degradation. We established a protocol for immunocytochemistry before FACS to prevent RNA degradation. Cells were fixed with a methanol-based fixative (UM-Fix), then subjected to immunocytochemistry. The addition of RNase inhibitor and dithiothreitol (DTT) to some buffers used for immunocytochemistry increased RNA integrity after cell recovery. We found increased copy numbers of mRNA in recovered cells using quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis. When RNase inhibitor and DTT were added, amplification of mRNA using T7 promoter was possible with RNA extracted from recovered cells after FACS. Our protocol ensures high quality RNA in cells recovered by FACS; therefore, gene expression analysis with a smaller number of cells is possible using pre-amplification of mRNAs. Our protocol for immunocytochemistry also might be applicable to RNA recovery after immunostaining.

Risk of surgical glove perforation in oral and maxillofacial surgery.

Oral and maxillofacial surgery, which involves several sharp instruments and fixation materials, is consistently at a high risk for cross-contamination due to perforated gloves, but it is unclear how often such perforations occur. This study aimed to address this issue. The frequency of the perforation of surgical gloves (n=1436) in 150 oral and maxillofacial surgeries including orthognathic surgery (n=45) was assessed by the hydroinsufflation technique. Orthognathic surgery had the highest perforation rate in at least 1 glove in 1 operation (91.1%), followed by cleft lip and palate surgery (55.0%), excision of oral soft tumour (54.5%) and dental implantation (50.0%). The perforation rate in scrub nurses was 63.4%, followed by 44.4% in surgeons and first assistants, and 16.3% in second assistants. The odds ratio for the perforation rate in orthognathic surgery versus other surgeries was 16.0 (95% confidence interval: 5.3-48.0). The protection rate offered by double gloving in orthognathic surgery was 95.2%. These results suggest that, regardless of the surgical duration and blood loss in all fields of surgery, orthognathic surgery must be categorized in the highest risk group for glove perforation, following gynaecological and open lung surgery, due to the involvement of sharp objects.

Massive immature teratoma in a neonate.

Teratomas are embryonal tumours composed of diverse tissues from three germinal layers with variable levels of maturity. The authors report a female patient with a large immature epignathus. Prenatal diagnosis permitted a caesarean section and tracheotomy to be planned under ex utero intrapartum treatment for airway obstruction. The tumour was successfully controlled, even though it was impossible to distinguish from normal tissue because it lacked a pedicle and capsule, using multidisciplinary therapy, including a series of surgical treatments and adjuvant chemotherapy. This case suggested that the level of serum alpha fetoprotein might be a useful indicator after surgery. At present, after 4 years, no regrowth has been observed and the patient has no problems with respiration or swallowing.

Seroepidemiology of Chlamydia pneumoniae in Japan between 1991 and 2000.

AIM: To clarify the endemic and epidemic status of Chlamydia pneumoniae in Japan, the incidence of anti-C pneumoniae antibody was evaluated over a period of 10 years. METHOD: Serum samples were collected from 4756 healthy individuals aged 6 months to 88 years (2488 male and 2268 female individuals) between 1991 and 2000. The antibody titre was determined by a microimmunofluorescence test. RESULTS: After stratification by age and sex in each year, distinct peaks with prevalences of 73.3% and 73.0% were noted in 1993 and 1999, respectively. The lowest prevalence rate was seen in 1996 (59.0%). The epidemic cycle has been estimated to be almost six years in this geographical area. CONCLUSIONS: Chlamydia pneumoniae infection is highly endemic in Japan, as it is in Western countries, and there is a year to year variability. Long term studies in Japan are needed to clarify the epidemic occurrence of C pneumoniae infection.

[A case of Coxiella burnetii pneumonia in an adult].

A 49-year-old man visited our hospital complaining of a continuous high-grade fever and cough which had appeared during his stay in Indonesia. He was admitted on the same day because his laboratory data showed marked inflammatory changes and his chest radiograph revealed an infiltrative shadow in the right upper lung field. Initial treatment with beta-lactams was not effective and both his symptoms and his chest radiograph worsened. However, treatment with erythromycin clearly had an effect. Then, we carried out several tests for detection of atypical pathogens including Mycoplasma and Chlamydia. Finally, the case was diagnosed as one of Coxiella burnetii pneumonia because the DNA of C. burnetii was detected from his sera and seroconversion of C. burnetii--specific antibody was observed among paired serum samples. C. burnetii is one of the most commonly recognized pathogens among community-acquired pneumonias in Western countries, but in Japan, reports of community-acquired C. burnetii pneumonia have been rare. This difference may be due to the features of Q fever, in which there are large differences in frequency and form from country to country and among areas of the same country. Surveillance of C. burnetii pneumonia in Japan and different area will be required.

In vitro activity of telithromycin, a new ketolide, against Chlamydia pneumoniae.

The in vitro activity of telithromycin, a new ketolide, was compared with those of roxithromycin, azithromycin, clarithromycin and erythromycin A against 20 strains of Chlamydia pneumoniae. The MICs and minimal chlamydiacidal concentrations of telithromycin for the 20 C. pneumoniae strains both ranged between 0.031 and 0.25 mg/L. Telithromycin was twice as active as roxithromycin, azithromycin and erythromycin A, but less active than clarithromycin. These results appear to indicate that telithromycin is an effective antibiotic that should play some role in the treatment of respiratory tract infections caused by C. pneumoniae.

The 7.5-kb common plasmid is unrelated to the drug susceptibility of Chlamydia trachomatis.

Using a new plaque cloning technique, we obtained unique Chlamydia trachomatis strains, which were confirmed to be free of the 7.5-kb common plasmid and glycogen in inclusions. The inclusions in plasmid-free and glycogen-negative strains were characterized by a target-like bull's eye morphology that appeared with the formation of a central translucent area that began to be seen clearly at 30 h postinoculation. The clear zone was composed of gel materials originating from reticulate bodies (RBs) that were abnormally large in size and irregular in shape, although one could not differentiate the aberrant RBs from normal RBs at early stages of development. The in-vitro susceptibility of these strains to various chemotherapeutic antibiotics was tested by comparison with their parent strains possessing the common plasmid. No difference was detected for any of the antibiotics tested, indicating that the 7.5-kb common plasmid is unrelated to the drug susceptibility of C. trachomatis.

Serum antioxidant micronutrient levels in oral lichen planus.

The aim of this study was to elucidate any association between oral lichen planus (OLP) and serum antioxidant micronutrients by a population-based case-control study. A total of 9536 subjects were examined, and 62 (58 reticular, 4 atrophic and erosive) diagnosed with OLP at referral facilities were compared with four controls per case (n=248) selected among disease-free subjects matched for age and sex. Serum levels of micronutrients (retinol, alpha-tocopherol, zeaxanthin/lutein and cryptoxanthin, lycopene, alpha-carotene and beta-carotene) were estimated by the high performance liquid chromatography (HPLC) method. Among cases, mean serum retinol level (2.820+/-0.849 micromol/l) was significantly higher compared with that of controls (2.562+/-0.735 micromol/l) (P<0.05). No significant differences were noted in carotenoid levels examined in this study, except for a lower level of lycopene found in atrophic/erosive OLP cases. The results of this study suggest that low serum retinol or carotenoid levels are not risk factors for occurrence of lichen planus, and any specific benefits of antioxidant micronutrients cannot be claimed for this inflammatory disorder.

Prevalence of asymptomatic infection with Chlamydia pneumoniae in subjectively healthy adults.

STUDY OBJECTIVE: To investigate the prevalence of asymptomatic infection with Chlamydia pneumoniae in subjectively healthy adults. DESIGN: Prospective study. SETTING: Kawasaki Medical School Hospital in Kurashiki, Japan. PARTICIPANTS: Total of 1,018 asymptomatic healthy adults (age range, 22 to 50 years; mean age, 32.4 years; 482 men and 536 women). MEASUREMENTS AND RESULTS: Nasopharyngeal swab specimens were obtained from all subjects and analyzed by isolation in cell cultures and polymerase chain reaction (PCR) test for C pneumoniae. Serum samples were also obtained and tested for C pneumoniae-specific antibodies by the microimmunofluorescence test. Of 1,018 specimens tested, 14 specimens (1.4%) were judged positive by culture and/or the PCR. Thirteen specimens were PCR positive, and 4 specimens were culture positive. There were three specimens positive by both tests and 11 specimens positive with discrepancies in culture and PCR results. None of the individuals met the serologic criteria for acute infection. Of 1,018 sera tested, 64.1% of men and 58.0% of women had antibody to C pneumoniae. The overall prevalence of antibody was 60.9%. Forty individuals (3.9%) had an IgG titer of > or = 1:512 or IgM titer of > or = 1:16. CONCLUSIONS: Our data suggest that asymptomatic infection with C pneumoniae may occur in subjectively healthy adults.

Serum antioxidant micronutrients and the risk of oral leukoplakia among Japanese.

A population-based case-control study was designed for the investigation of any association between serum micronutrient levels and oral leukoplakia. Out of a total of 9536 subjects over the age of 40 years who participated in the oral mucosal screening programme in Tokoname city, 48 cases detected with oral leukoplakia (38 male:10 female) were recruited. For each case, four controls matched by age and sex were selected from the same cohort. We examined the fasting serum levels of retinol, alpha-tocopherol, zeaxanthin and lutein, cryptoxanthin, lycopene and carotenoids (alpha-carotene and beta-carotene) by high-performance liquid chromatography. Among males with leukoplakia mean serum lycopene and beta-carotene levels (0.175+/-0.202, 0.357+/-0.295 micromol/l) were significantly lower than those of controls (0.257+/-0.252, 0.555+/-0.408 micromol/l) (P<0.05, P<0.005). Logistic regression analysis with leukoplakia as the dependent variable showed that high serum levels of beta-carotene were related to low risk of oral leukoplakia (odds ratio 0.160, 95% C.I.: 0.029-0.866, P<0.05). There were no significant differences in any of the serum nutrients estimated in female subjects. Our results suggest for the first time that high serum levels of beta-carotene may provide protection against oral precancer for the Japanese male.

Outcome following a population screening programme for oral cancer and precancer in Japan.

The purpose of this study was to evaluate the validity and outcome of an oral mucosal screening programme conducted in Japan. All adults over the age of 40 years resident in Tokoname city were invited by letter to attend a free general health screen and oral examinations annually. Twenty- to 39-year-old females were also encouraged to participate. During the years 1996-98, a total of 19056 subjects (5885 male, 13171 female: mean age 60.7+/-11.3 years) were examined by three types of screeners: postgraduate dental residents (n=17), hospital dentists (n=5) and general dental practitioners (n=15). In the cohorts examined screening dentists recorded oral mucosal lesions in 783 (4.1%) subjects. Of those detected with mucosal lesions, 200 (25.5%) were referred and 137 (68. 5%) attended for follow up examination in hospital departments by specialists with full diagnostic back up facilities. Thirty-nine subjects were confirmed as having oral cancer or precancer (two squamous cell carcinomas, 37 leukoplakias) and 40 with lichen planus. Five false positives (3.6%) were found. Among the cases referred under other diagnostic labels no cancers were detected. Among those who attended for confirmation of oral cancer or precancer by a specialist, sensitivity, specificity and positive predictive value were 0.92, 0.64 and 0.78, respectively. In males with oral cancer/leukoplakia, the prevalence of smoking, drinking and combined habits was higher than those with lichen planus or among negative subjects. The sensitivity and specificity determined from this research suggests that the performance of the Japanese dentists employed in screening was satisfactory. An attendance of 68% for re-examination by specialists compares well with other reported studies measuring patient compliance.

Metallothionein immunoreactivity in head and neck carcinomas; special reference to clinical behaviors and chemotherapy responses.

Metallothionein (MT), has selectively binding affinity for heavy metal ions and over expression of MT has a potential against resistance for CDDP anticancer agents and radiation treatment. The role of MT immunoreactivity of squamous cell carcinoma in oral and pharyngeal regions (n = 28) and in the maxillary sinus region (n = 3) was evaluated for distribution patterns of MT and clinicopathologic behaviors. All the sections were examined in 400x and counted for MT positive cells over 5 fields of tumor growing foci. MT immunoreactivity was expressed in both tumor cell cytoplasm and nuclei, and showed heterogeneous localization in tumor epithelial cells and in the stroma. Immunohistochemical localizations showed mosaic patterns as the highest MT staining tumor cells intermingled with negative or low staining cells in neoplastic foci, and in stromal cells. Histiocytic and fibrocytic cells in both peripheral and interstitial stromas were also not stained homogeneously. In oral and pharyngeal carcinomas (n = 28), MT positive cell index in treated cases (n = 11) was 17.85% and that in non treated tumors (n = 17) was 25.19%. In maxillary sinus carcinomas (n = 3), MT positive index was 4.56% and showed lowers levels as compacted to other SCC sites. Among histological grading in oral and pharyngeal SCCs, MT index of well differentiated SCC (n = 9) was 17.04%, of moderately differentiated SCC (n = 13) 21.92% and poorly differentiated SCC (n = 6) was 31.06%. There is no significant correlation of positive index of metallothionein between treated and untreated samples taken in oral and pharyngeal SCCs.

Oral cancer screening as an integral part of general health screening in Tokoname City, Japan.

OBJECTIVES: To measure the attendance and compliance rates in oral mucosal screening (OMS) offered as part of a general health screen (GHS) undertaken as an organised programme in Japan. METHODS: In 1996, all adults over the age of 40 years resident in Tokoname City were invited by letter to attend a free GHS annually, conducted by the municipal cooperation and the medical and dental societies of Tokoname City. In the later years only those who attended in 1996 were reinvited. Females aged less than 39 years were also allowed to attend if they wished to participate in the GHS. The GHS consisted of completion of a self administered questionnaire to identify past and current illnesses and any medications used by the screened population coupled with a routine physical check, chest x ray, ECG, and blood and urine examination. The GHS was programmed annually during the years 1996 to 1998. All those attending the GHS were invited to participate in an OMS conducted under the same roof by a visiting dentist (n=37). A referral pathway was established for screen positives requiring follow up. RESULTS: A total of 19 305 subjects (5955 males, 13350 females; mean age 59.2 years; 7033 in 1996, 6289 in 1997, and 5983 in 1998) attended the GHS. Of those who attended the GHS, 19 056 (98.7%) attended the OMS (1.3% refused). This rate was fairly constant over the three years. Excluding repeat examinations, new cases recruited for OMS over the three years were 8723. Of those participating in OMS, 4269 (60.7%) attended all three years. In the cohorts examined, screening dentists recorded oral mucosal lesions in 5.4% in 1996, in 4.0% in 1997, and in 2.6% in 1998. Overall, this amounted to a positive detection rate of 4.1%, or 4.9% excluding repeat examinations. A higher prevalence of oral pre-cancer was recorded among male smokers. CONCLUSIONS: The overall results suggest that although compliance with attending a free GHS was low (26.2%) among Japanese subjects over 40 years of age, of those who complied 74-76% reattended annually. Hence a satisfactory participation rate can be obtained in Japan for OMS when this is coupled to a GHS conducted at the same visit.

Ribosomal RNA supplementation highly reinforced cell-free translation activity of wheat germ.

We have constructed an inexpensive, highly efficient eukaryotic cell-free translation system. Wheat germ rRNA (WG rRNA) was prepared by phenol/chloroform (P/C) extraction, a simple and quick method, from wheat germ, an inexpensive and commercially available by-product of flour production. Addition of a small amount of WG rRNA into a wheat germ cell-free translation system increased the protein productivity of the system 6- to 8-fold. Isolated 18S or 28S rRNA alone enhanced the protein production only 2-fold or 3.9-fold, respectively, at maximum. On the other hand, their equimolar mixture enhanced the production as much as the whole WG rRNA, indicating 18S and 28S rRNA synergistically functioned to enhance protein synthesis. Addition of WG rRNA slightly improved the stability of mRNA in the cell-free translation system, which explained only partly the enhancement of protein production. Addition of WGE or ribosome containing approximately the same amount of rRNA in the form of protein-rRNA complex as WG rRNA added to the system did not increase the protein production in the translation system. When ribosome in the cell-free translation system was replaced with WG rRNA, the system did not exhibit any detectable translation activity, indicating that the translation activity of WG rRNA is negligible in comparison with that of ribosome. These results indicated that WG rRNA affected some mechanisms regulating the translation rate in wheat germ cell-free system, resulting in increased protein production.

Depth of invasion as a predictive factor for cervical lymph node metastasis in tongue carcinoma.

BACKGROUND: Many histopathologic parameters in head and neck squamous cell carcinoma have been identified as predictive factors for cervical metastasis. Several studies focused on tumor thickness, and the depth of invasion was suggested to have a relationship to the occurrence of cervical metastasis. Nonetheless, the criterium for elective neck therapy in terms of tumor depth is still inconclusive. Therefore, a retrospective study was undertaken to substantiate the differing results in the literature with our own findings concerning the interrelationship between tumor thickness and clinically suspicious neck, as well as occult neck disease. In addition, the study attempted to identify further predictive factors for cervical metastasis in squamous cell carcinoma of the tongue. METHODS: The medical records of 34 primary tongue carcinomas operated without any preoperative therapy between 1980 and 1991 were reviewed. Each patient's tumor type, tumor location, tumor size, invasion mode, depth of invasion, intralymphatic tumor emboli, and perineural invasion were evaluated. Chi-square contingency tables were used to correlate clinical or histopathologic parameters with metastasis in the neck. RESULTS: The overall cervical metastatic rate was 35.3% (n = 12/34). In univariate analysis, invasion mode and depth of invasion were statistically significant predictors of regional metastasis at p = 0.0019 and p = 0.0003, respectively. In the group in which tumor depth exceeded 5 mm, the metastatic rate was 64.7% (11/17). In contrast, when the depth of invasion was less than 5 mm, the incidence of cervical metastasis was 5.9% (1/17). Clinically negative necks turned out pathologically positive in 30.0% (n = 9/30). The tumor depth exceeded 5 mm in 43.4% (13/30) of the N0 necks. In the conversion (N0-->N1) group, tumor depth exceeding 5 mm was noted in 88.9% (n = 8/9). CONCLUSIONS: We suggest that there is a discerning point at 5 mm of tumor depth at which cervical metastasis is probable. Electric neck therapy (surgery or irradiation) is strongly indicated for tumors exceeding 5 mm invasion. Tumor invasion mode and tumor size also bear an impact on the indication for therapy.

Annual screening for oral cancer and precancer by invitation to 60-year-old residents of a city in Japan.

Since 1986, annual screening for oral cancer and precancer by postal invitation has been undertaken among 60-year-old residents of Tokoname city, Japan. Clinical examination of the oral soft tissues is performed by groups of four different, specially trained general dental practitioners in the city health centre on one day each year. Following screening each subject receives an individual consultation with an oral medicine specialist. Individuals considered to need full diagnostic follow up or treatment are advised to attend a secondary care referral facility. Between 1986 and 1993, 802 out of 5187 eligible residents (15.5 per cent) were screened of whom 38 (4.7 per cent) were designated by the screeners as positive for oral cancer, erythroplakia, leukoplakia, lichen planus or chronic candidosis. Of these, 32 were referred and 25 attended for follow up examination in hospital departments by specialists with full diagnostic back up facilities. Twenty subjects were confirmed as having a potentially malignant lesion. For these referred patients only, the proportion of correct decisions by the screeners out of all oral cancer/precancer screening decisions was 81 per cent. Positive and negative predictive values for those attending for follow up were 0.80 and 0.82 respectively. Ways of improving the effectiveness and adequacy of the programme are considered.

Prevalence study of oral mucosal lesions in a selected Cambodian population.

The aim of this study was to investigate the epidemiology of oral mucosal lesions in a selected Cambodian population to obtain pilot data useful in planning an oral health data base for the country. Due to unstable conditions in Cambodia, the validity of population data related to present census information is highly questionable. Therefore, prior to this investigation a census registration was carried out using local health workers as registrars in nine villages of a commune. In the period July 4-31, 1991, a total of 1319 individuals (953 women, 366 men, 15-99 yr) were examined by one oral surgeon in the nine villages of Kok Trop Commune, Kandal Stung District, southwest of the capital Phnom Penh. Clinical diagnoses were based on WHO criteria. Information on smoking habits, betel nut chewing habits, and alcohol use was collected by 4 Khmer dental personnel. In total, 71 lesions were recorded in 64 (4.9%) individuals. Leukoplakia was found in 1.1%, lichen lesions in 1.8%, candidosis in 1.4%, submucous fibrosis in 0.2%, cancer in 0.1% and other diagnoses in 0.8%. The prevalence of leukoplakia was 2.2% and 0.6% among men and women respectively, a statistically significant difference (P < 0.05). There were significantly more smokers (P < 0.01) among subjects with leukoplakia (64.3%) than among those without this lesion (28.6%). All subjects with lichen lesions were women. The age-adjusted relative risk for developing lichen among betel nut chewers as compared to non-chewers was 3.3.(ABSTRACT TRUNCATED AT 250 WORDS)