Neural network application in Japanese sign language: distinction of similar Yubimoji gestures.

In a previous paper, the authors built a neural network model to recognize Japanese sign language syllabary or yubimoji. One of the problems encountered in that study was the accurate digital representation and distinction of similar yubimoji gestures, i.e. gestures with the same finger flexure positions but with different hand/finger orientations. This study focuses on these yubimoji gestures. Using data from a glove interface with bend sensors and accelerometers, a neural network was built, trained and tested. The network performed well and good results were obtained.

Measurement of urinary zearalenone concentrations for monitoring natural feed contamination in cattle herds: On-farm trials.

The aims of the present study were to investigate the efficacy of measuring bovine urinary zearalenone (ZEN) concentrations by using a commercially available ELISA method in cattle kept under different feeding conditions to monitor the natural contamination of feeds at the farm level, and to investigate the effects of supplementation of a mycotoxin adsorbent (MA) product in the feed based on urinary ZEN concentration. First, Japanese Black cattle herds kept for breeding (4 herds) and fattening (4 herds) purposes were provided with similar feeding conditions. Then, urinary samples from 5 cows in each herd were collected and analyzed. Second, dairy cows from 1 herd fed with total mixed rations (TMR) were selected. After thorough mixing of the MA (40 g/d) with TMR, the supplemented TMR was fed according to the following schedule: with MA for 2 wk, without MA for 3 wk; then with MA for 2 wk and without MA for 6 wk. Urine samples were collected from cows (n = 6 to 7) and examined before and after each interval. Zearalenone concentrations were measured by the ELISA and liquid chromatography-tandem mass spectrometry methods. The concentration of ZEN and its metabolites was expressed after creatinine (Crea) correction [ZEN or metabolites (pg/mL)/Crea (mg/dL); pg/mg of Crea]. In the first experiment, the urinary concentrations of ZEN and its metabolites were variable in all herds, and significant differences were observed between herds. In 1 fattening herd, in particular, urinary ZEN concentrations were greater (P < 0.001) than in the other 3 herds. This might reflect significant natural ZEN contamination of the feed at the farm level. In Exp. 2, urinary ZEN concentrations displayed peculiar trends after supplementation with MA. After 2 wk of supplementation, a significant decrease of ZEN (P < 0.05) was observed. Zearalenone concentrations remained at a reduced amount during 3 wk without MA supplementation and 2 wk with MA supplementation. When MA was not added to the feed for the next 6 wk, the concentrations increased to the original quantity. These findings indicate the usefulness of measuring concentrations of urinary ZEN and its metabolites not only for monitoring the natural ZEN contamination of cattle feed at the farm level but also for in vivo evaluation of MA function after supplementing feeds with MA.

Recognition of Japanese finger spelling gestures using neural networks.

Effective communication with the hearing and speech impaired often requires at least a basic working knowledge of sign language gestures, without which a memo pad and pen, or a mobile phone's notepad is indispensable. The aim of this study was to build a neural network that could be used to recognize static finger-hand gestures of the yubimoji, the Japanese sign language syllabary. To build the network, signal inputs from a data glove interface were taken for each of the static yubimoji gestures. The network was trained and tested 10 times using a multilayer perceptron model. Overall, only 18 of the 41 static gestures were successfully recognized. One of the reasons was attributed to the inability of the data glove to measure gesture directions particularly for yubimoji gestures with similar finger configurations. Future work will focus on these problems as well as the inclusion of dynamic yubimoji gestures.

An outbreak of psittacosis in a bird park in Japan.

An outbreak of psittacosis related to a bird park occurred in Matsue City, Shimane Prefecture, Japan, during winter 2001. Seventeen cases of psittacosis (12 visitors, three staff, and two student interns) were confirmed. A cohort study was conducted among the park staff and students to determine the risk factors for the development of acute serologically confirmed psittacosis (SCP) infection. Being 'bird staff' had an increased risk of SCP infection (RR 3.96, 95% CI 1.48-10.58). Entering the staff building, where ill birds were maintained without proper isolation, was also associated with an increased risk of SCP infection (RR 3.61, 95% CI 1.03-12.6). Isolation of ill birds and quarantine measures were found to be insufficient. Dehumidifiers and a high-pressure water spray under a closed ventilation environment may have raised the concentration of Chlamydophila psittaci in the hothouses. Bird park staff and visitors should be educated about psittacosis.

New criteria for immunofluorescence assay for Q fever diagnosis in Japan.

A study was made to evaluate the cutoff value of indirect immunofluorescent-antibody (IFA) test for Q fever diagnosis in Japan. We used 346 sera, including 16 from confirmed Q fever cases, 304 from Japanese pneumonia patients, and 26 from negative cases. Thirteen sera from the confirmed Q fever cases with an immunoglobulin M (IgM) titer of > or =1:128 and/or IgG titer of > or =1:256 by the IFA test were positive by both enzyme-linked immunosorbent assay (ELISA) and Western blotting assay (WBA), whereas 298 sera from pneumonia patients and 26 negative sera with an IgM titer of < or =1:16 and an IgG titer of < or =1:32 by the IFA test were negative by both ELISA and WBA. In the proposed "equivocal area," with an IgM titer of > or =1:32 and < or =1:64 and/or an IgG titer of > or =1:64 and < or =1:128, we found 9 sera, 3 from confirmed Q fever cases and 6 from Japanese pneumonia patients, by the IFA test. Three sera from the confirmed Q fever cases and one of the sera from pneumonia patients were IgM and/or IgG positive by both ELISA and WBA. These results suggest that a single cutoff value for the IFA test may cause false-positive and false-negative results. In conclusion, this study showed that an "equivocal area" should be used for the IFA test rather than a single cutoff value and that sera in the equivocal area should be tested by additional serological assays for confirmation.

Inactivation of Chlamydia trachomatis and Chlamydia (Chlamydophila) pneumoniae by ozone.

AIMS: To clarify the inhibitory effects of ozone on Chlamydia trachomatis and C. pneumoniae. METHODS AND RESULTS: Cell culture was performed using HeLa229 cells for C. trachomatis, and Human Line cells for C. pneumoniae. C. trachomatis strain D/UW-3/Cx and C. pneumoniae strain AR-39 were used. Ozone water was generated by an ozone water dispenser and diluted to desired concentration just before each experiment. Preinoculation minimum cidal concentration (MCC) and postinoculation MCC methods were employed. In preinoculation MCC, chlamydial strains were treated with serially diluted ozone water followed by inoculation to cells. In postinoculation method, chlamydial strains were inoculated to cells and incubated for 24 h. Then infected cells were treated with ozone water, followed by additional incubation for 48 h. Complete inactivation was obtained in preinoculation MCC method at 0.5 ppm of ozone water for 30 s, or 4 ppm for 5 s. CONCLUSION: Ozone at a concentration of 4 ppm was enough for immediate inactivation of both C. trachomatis and C. pneumoniae. SIGNIFICANCE AND IMPACT OF THE STUDY: Ozone water at 4 ppm should be applicable for prevention of C. trachomatis urogenital infections.

Suppressive effects of breast milk on oxidative DNA damage in very low birthweight infants.

BACKGROUND: Human milk contains many kinds of antioxidant and is considered to prevent diseases mediated by oxygen free radicals in very low birthweight (VLBW) infants. AIMS: To examine the antioxidant effects of breast milk in VLBW infants by determining urinary 8-hydroxydeoxyguanosine (8-OHdG) excretion, which is known to be a non-invasive marker for in vivo oxidative DNA damage. METHODS: Urinary 8-OHdG concentrations were measured in 15 breast fed and 14 formula fed VLBW infants at 2, 7, 14, and 28 days of age. RESULTS: Urinary 8-OHdG excretion at 14 and 28 days of age was significantly lower than at 2 and 7 days of age in the breast fed group, and significantly lower than in the formula fed group. CONCLUSION: This is the first direct evidence of the antioxidant action of human milk in VLBW infants.

Lack of plasma leptin response to feeding in newborn infants.

OBJECTIVE: Previous reports of the postprandial regulation of leptin are controversial, and there have been few studies on the effects of breast-feeding on postprandial regulation in newborn infants. We examined the response of plasma leptin to breast- and formula-feeding in newborn infants. METHODS: We measured the plasma leptin levels using an enzyme-linked immunosorbent assay kit before and after feeding in 12 breast-fed and 11 formula-fed mature infants. RESULTS: There was no significant difference in plasma leptin levels in breast-fed infants before and after feeding or in artificially fed infants before and after feeding. CONCLUSION: Our results suggest that feeding does not play a role in the acute response of circulating leptin levels in either breast- or formula-fed infants.

[Tsutsugamushi disease (scrub typhus) in Japan: epidemiological aspects].

Epidemiological aspects of tsutsugamushi disease (scrub typhus) in Japan in 1998 were analyzed using questionnaires. Four hundred and sixteen scrub typhus cases were reported in 24 prefectures in 1998. The annual number of the patients in 1998 was similar to those in the preceding three years. There was no sex difference. The patients at the age of 51 or greater accounted for 72% of the total cases. Patients engaged in farming and forestry accounted for 32% and 14% cases, respectively. Fifty-six, 21 and 19% were reported in Kyusyu, Kanto and Tohoku-Hokuriku districts, respectively, 96% of the total cases being reported in these 3 districts. Most cases were reported from April through June with some from October through December in Tohoku-Hokuriku districts, while most cases were reported from October through December in other districts, including Kyusyu and Kanto districts. Thus, there was a difference in epidemic seasons among the districts. Serotypes of scrub typhus rickettsia were analyzed by serum antibody titers in the Kyusyu district. The novel Kawasaki and Kuroki types were major strains; however, no geographical difference was seen within the Kyusyu district. Interestingly, 24 cases were diagnosed only by the new serotypes not by the classical serotypes (Kato, Karp and Gilliam) in serological tests. This result suggests that further investigations are required to determine the prevalent serotypes in each district and to improve the serological tests. This was the first comprehensive report of epidemiology of scrub typus in entire Japan. Information obtained in the present study provides deep insight into prediction, diagnosis, treatment and prevention of scrub typhus in Japan.

[Tsutsugamushi disease (scrub typhus) in Japan: clinical features].

Clinical features of tsutsugamushi disease (scrub typhus) were analyzed, based on 416 cases reported in Japan in 1998. Three major clinical symptoms: eschar, fever and rash were found in 87%, 98% and 92% of the cases, respectively. Elevated levels of CRP, GOT, GPT and LDH were observed in 96%, 85%, 78% and 91%, respectively. These clinical and laboratory findings were observed in the majority of the cases and considered important for diagnosis. Disseminated intravascular coagulation developed in 21 cases, indicating that scrub typhus can be life threatening. Lymphadenopathy was observed in 51% of the cases. Enlarged lymph nodes were limited to the local sites in 75% of these lymphadenopathy cases and most of these sites were adjacent to eschars. Most eschars were scabbed and located in the abdomen and the lower half of the body, especially the feet. This suggests that these parts are frequently exposed to tsutsugamushi mites. Furthermore, the skin is soft in these parts and covered by cloth. These factors may make it possible for mites to keep biting without being noticed for several hours, long enough for rickettsial transmission. Interestingly, eschar and rash were absent in 14% and 8% of the cases, respectively. This result suggests that the cases without the unique symptoms may have been misdiagnosed as common cold or other febrile illnesses. One hundred and fifty-four suspected cases were not scrub typhus cases by the serological tests. The three major clinical symptoms were present in approximately a half of these negative cases, eschar being observed in approximately 70%. This may suggest the presence of new type of scrub typhus can not be diagnosed by the present laboratory tests. Clinical features of scrub typhus in Japan were well revealed, and information obtained in the present study is useful for improving clinical diagnosis. It should, however, be stressed that there were cases that could not be correctly diagnosed only by the clinical symptoms, suggesting that it is important to improve the serological tests.

[Preliminary results of PET activation study in cerebral arteriovenous malformation (AVM), using C15O2 and 18F-FDG].

The authors applied PET activation study to two patients with arteriovenous malformation (AVM) to localize primary motor cortex before surgery or embolization. The change in regional cerebral blood flow (rCBF) was measured during foot movements in Case 1 who had a 2-cm AVM located in the post-central gyrus. Superimposed PET/MRI images revealed that the rCBF increase was located in the pre-central gyrus. Its validity was confirmed by intraoperative cortical mapping using electrical median nerve stimulation. The patient safely underwent total removal of AVM. The change in regional cerebral metabolic rate for glucose (rCMRglc) was measured during hand movements in Case 2 who had a huge AVM over the central sulcus. Superimposed PET/MRI images revealed that hand movements significantly increased rCMRglc in the frontal cortex, which was separated from the original primary motor area. The patient safely underwent partial embolization, although he suffered transient weakness of the face after embolization. The preliminary results strongly suggest that PET activation study is useful to localize precisely cortical functions of the patients with AVM, thus reducing morbidity after treatment. The results also suggest that cortical functions may undergo translocation when huge AVM involves the eloquent area.

In vitro analysis of the change in resistance of Chlamydia trachomatis under exposure to sub-MIC levofloxacin for a therapeutic term.

BACKGROUND: While fluoroquinolone-resistant Chlamydia trachomatis strains have not been clinically isolated, they were isolated in an in vitro study recently. METHODS: To determine whether C. trachomatis strains develop resistance under sub-MIC antibacterial exposure in a clinical therapeutic term, C. trachomatis strains were exposed to sub-MIC levofloxacin (LVFX) for about 2 weeks. The MIC of LVFX was measured and DNA fingerprinting was performed every 72 h by PCR using random primers. RESULTS: There was almost no change in the MIC under exposure to 0.125 microg/ml LVFX. However, some mutational changes in DNA fingerprints developed. CONCLUSIONS: In clinical therapeutic terms, resistant strains of C. trachomatis will probably not develop, even if sub-MIC LVFX is employed.

[Effect of heparin cofactor II on the antithrombin III activities measured by thrombin methods or factor Xa methods--fundamental studies and clinical studies using the plasma of pregnant women].

We evaluated the effects of heparin cofactor II(HC II) on the antithrombin III(AT III) activities measured by the methods of thrombin or factor Xa. Reagents A and B were using the method of thrombin and reagent C was based on the method of Xa. Purified HC II was directly measured or indirectly measured after the dilution with control plasma. Cross reaction of HC II in AT III assay were negligible in reagent C, but substantial amount of AT III activities were measured in reagent A and B. Plasma AT III activities from full-term pregnant women were significantly higher than those from non pregnant control women in reagent A, but comparable in reagent B or C. These results indicate that AT III activities measured by thrombin methods by thrombin were overestimated in pregnant women due to the cross-reactivities of HC II. It is recommended that AT III activities would be measured by the methods of factor Xa.

The role of neurosecretory neurons in the pars intercerebralis and pars lateralis in reproductive diapause of the blowfly, Protophormia terraenovae.

Microlesions of the brain were made to examine the role of neurosecretory neurons in the pars intercerebralis (PI) and pars lateralis (PL) in the induction of reproductive diapause of the female blowfly Protophormia terraenovae. Under both diapause-inducing (LD 12:12, 20 degrees C) and diapause-averting conditions (LD 18:6, 25 degrees C), the ovaries invariably failed to develop when the PI was removed. When the PL was removed bilaterally, the ovaries developed in most of the females, irrespective of the rearing conditions. Removal of the PL prevented females from entering reproductive diapause. These results show that certain neurosecretory neurons in the PI are necessary for vitellogenesis, and that the PL contains inhibitory neurons which suppress vitellogenesis during reproductive diapause.

Neurons projecting to the retrocerebral complex of the adult blow fly, Protophormia terraenovae.

Anatomical study of neurons projecting to the retrocerebral complex of the adult blow fly, Protophormia terraenovae, was done by NiCl2 filling and immunocytochemistry. Retrograde filling through the cardiac-recurrent nerve labeled three groups of neurons in the brain/subesophageal ganglion: (1) paramedial clusters of the pars intercerebralis, (2) neurons in each pars lateralis, and (3) neurons in the subesophageal ganglion. The pars intercerebralis neurons send prominent axons into the median bundle and exit from the brain via the contralateral nervus corporis cardiaci. Based on the projection pattern, two types of the pars lateralis neurons can be distinguished: the most lateral pairs of neurons contralaterally extend through the posterior lateral tract and the remainder ipsilaterally extend through the posterior lateral tract. The neurons in the subesophageal ganglion run through the contralateral nervus corporis cardiaci. The dendritic arborization of the pars intercerebralis and pars lateralis neurons is restricted to the superior protocerebral neuropil and to the anterior neuropil of the subesophageal ganglion where the neurons in the subesophageal ganglion also project. Retrograde filling from the corpus allatum indicated that the pars lateralis neurons and a few pars intercerebralis neurons project to the corpus allatum, but that the neurons in the subesophageal ganglion do not. Orthograde filling from the pars intercerebralis and staining by paraldehyde-thionin/paraldehyde-fuchsin indicated that the pars intercerebralis neurons project primarily to the corpus cardiacum/hypocerebral ganglion complex. Immunostaining with a polyclonal antiserum against diapause hormone, a member of the FXPRLamide family, suggests that some of the subesophageal ganglion neurons contain FXPRLamide-like peptides.

[Serovar distributions of cervical Chlamydia trachomatis isolated in Nagano Prefecture].

Between 1992 and 1998, serotyping of 82 Chlamydia trachomatis cervical isolates were examined by micro-IF method in Nagano Prefecture. Of these, 17 isolates (20.7%) were serovar E, 15 (18.3%) were serovar F, 14 (17.1%) were serovar D and 10 (12.2%) were serovar G, and the isolates typed these serovars were found to be 68.3% whole. Furthermore, serovars B (7.3%) and K (9.8%) were comparatively frequently found. From 1992 to 1994, the number of isolates of B-complex, intermediate and C-complex were 18, 17 and 6, respectively. On the other hand, from 1995 to 1998, the number of the isolates were 19, 8 and 14, respectively. The distribution of serovars of C. trachomatis tended to fluctuate from serovars D and E to F and G as the patients grew older. Moreover, serovar E isolates were only detected from patients less than 40 years old. Although most of the serovar I isolates were detected from the patients with vaginal discharge, the other isolates did not clearly indicate the relationship of serovars and clinical symptoms.

Flow cytometric method for enumeration and classification of reactive immature granulocyte populations.

We developed a flow cytometric method for the enumeration and classification of nonmalignant immature granulocytes (IG). In this study, IG are defined as most immature (IG stage 1: promyelocytes and myelocytes) and as more mature (IG stage 2: metamyelocytes). Blood specimens from 46 patients with documented infectious or inflammatory disease and known presence of IG (by routine manual microscopy) were analyzed. For a reference manual differential count, we used a 400 white blood cell (WBC) differential and separated granulocytes into promyelocytes and myelocytes combined, metamyelocytes, and included band cells in the mature, segmented neutrophil population. The flow cytometric method is based on three-color staining of whole, anticoagulated blood with CD45-PerCP, CD16-FITC, and CD11b-PE-labeled monoclonal antibodies and a three-step gating procedure. The flow cytometric results were confirmed by cell sorting and microscopic evaluation of the sorted cells. A total of 10,000 events, excluding debris, were recorded per specimen and IG stage 1 (CD16-/CD11b-), IG stage 2 (CD16-/CD11b+), and mature neutrophils (CD16+/CD11b+) were categorized. Regression and correlation between flow cytometric IG and the manual differential showed y = 1.34x + 0.95, r(2) = 0.86 for IG stages 1 and 2 combined versus promyelocytes, myelocytes, and metamyelocytes. For IG stage 1 versus microscopic counts of promyelocytes and myelocytes, the results were y = 1.53x + 1.24, r(2) = 0.76; for IG stage 2 versus manual metamyelocyte count, y = 0.77x + 0.21, r(2) = 0.58. Reproducibility of the flow cytometric method showed a coefficient of variation (CV) of 6.8% for all IG combined compared with a CV of 50.2% for manual differential IG count (based on a routine 100 WBC count). Samples were found stable at least 12 h at 25 degrees C and at least 48 h at 4 degrees C for flow cytometry. After staining and lysing, the sample was stable for at least 120 min at room temperature. We analyzed samples from patients with myelodysplastic and myeloproliferative disease separately. We found that CD16- mature neutrophils falsely elevated the flow cytometric IG count. Similar results were obtained in blood from patients treated with granulocyte-colony stimulating factor (G-CSF). Although this restricts the use of the method somewhat, we believe that this flow cytometric method is useful for enumerating reactive IG, as well as for evaluating automated methods for IG identification by hematology analyzers.

Detection of antimicrobial-treated Chlamydia trachomatis with Amplicor PCR test kit.

A polymerase chain reaction (PCR) method for the detection of Chlamydia trachomatis has been developed and is now available in the clinical setting. However, one of the major problems with this method is possible false-positive detection posttreatment. Nonviable C. trachomatis was established by in-vitro exposure to an antimicrobial agent, and we tried to detect the nonviable cells (NVCs) of C. trachomatis by PCR with variant primer sets. C. trachomatis strains (D/UW-3/Cx) were cultured in a medium containing the antimicrobial agent, at 8 x MIC (minimal inhibitory concentration) 15 to 20 h postinfection. Amplicor and two sets of PCR primers were used to detect the DNA of NVCs. Serial passages of NVCs were done five times. All samples were positive on Amplicor, and all except the fourth passage were positive for the two sets of primers. Although the PCR test appears to be valuable, NVCs may possibly be detected by this method, and this may be clinically responsible for the false detection of C. trachomatis after appropriate antimicrobial chemotherapy.

[Clinical reference values for laboratory hematology tests calculated using the iterative truncation method with correction: Part 2, Reference values for white blood cell (WBC) count, WBC differential including segmented neutrophil, band neutrophil, lymphocyte, monocyte, eosinophil, basophil, platelet count and mean platelet volume].

Age and sex dependent differences in the clinical reference values for white blood cell(WBC) count, WBC differential including segmented neutrophils(SEG), band neutrophils(BAND), lymphocytes (LYMPH), monocytes(MONO), eosinophils(EOSINO), basophils(BASO), platelet count(PLT) and mean platelet volume(MPV) were calculated by the iterative truncation method with correction(Usui's method) using the result from tests on 6,300 patient specimens obtained at Kyoto University Hospital. WBC counts exhibited log-normal distribution in all age either in male and female. PLT counts and MPV showed normal or log-normal distribution. WBC differential (SEG, BAND, LYMPH, MONO, EOSINO, BASO) usually showed log-normal distribution. As to the reference value for WBC, high limit values gradually decreased in the age range of 5-14 year old in males and females. After 15 year old, it gradually increased and reached adult's level. Low limit values for WBC gradually decreased with age both in males and females. The reference value for SEG showed similar change to WBC. The reference value for LYMPH gradually decreased age both in males and females. The reference values for BAND, MONO, EOSINO, BASO were generated by the percentile method instead of the iterative truncation method because of the strong deviation in the distribution pattern. BAND, MONO and BASO values were very stable for all ages in males and females. High limit values for EOSINO in the age range of 5-10 year old in males and females remained constant, and gradually decreased in 10-20 year old, and were stable at ages above 20 year old. PLT counts gradually decreased with age, and the MPV remained constant in all age populations both in male and female.