Prevalence of Bartonella infection in cats and dogs in a metropolitan area, Thailand.

The prevalence of Bartonella infection was studied in 312 cats and 350 dogs in the Bangkok metropolitan areas, Thailand, between June 2001 and February 2003. Bartonella was isolated from 47 (16.3%) of 288 stray cats, but from none of the 24 pet cats studied. Of the 47 Bartonella-positive cats, 45 animals were infected with only B. henselae, one was infected with only B. clarridgeiae, and one with both B. henselae and B. clarridgeiae. 16S rRNA typing showed that 40 cats were infected with B. henselae type I, four with B. henselae type II, and one with both B. henselae types I and II. These results indicated that B. henselae, especially type I, was prevalent in stray cats that constituted a large Bartonella reservoir in Bangkok. B. clarridgeiae was isolated for the first time in Asia from one of 350 dogs.

[A case of pulmonary alveolar proteinosis complicated with pneumoconiosis and myelodysplastic syndrome].

A 72-year-old man who had worked as a coal worker for 28 years and as a tunnel construction worker for 18 years was admitted because of fever, dyspnea, and appetite loss. Pneumoconiosis had been diagnosed when he was 64 years old and myelodysplastic syndrome at 71 years of age. After admission, the patient was treated with antibiotics and anti-fungal drugs but did not respond. Respiratory failure gradually worsened and he died. Autopsy specimens revealed that the cause of death was exacerbation of respiratory failure due to pulmonary proteinosis rather than pulmonary infection. This is a case of pulmonary proteinosis complicated with pneumoconiosis and myelodysplastic syndrome which, to our knowledge, is rare. We also considered the possibility that defective pulmonary macrophage function due to myelodysplastic syndrome and long-term silica inhalation played a part in the development of pulmonary alveolar proteinosis in this case.

[A case of lymphangioleiomyomatosis with carcinomatous lymphangiosis].

A 66-year-old woman had a history of partial gastrectomy and resection of the residual stomach because of early gastric cancer and its recurrence. She had been suffering from dyspnea on effort, hemosputum, and cough since the age of 52 years. Chronic pulmonary emphysema and bronchial asthma were diagnosed when she was 59. On January 31, 1996, she was admitted to UOEH hospital with a complaint of increased dyspnea. In spite of treatment with oxygen, steroid, and inhaled anti-cholinergic agent, her condition deteriorated. Subsequently, she had DIC, respiratory failure and reticulolinear opacities were seen on chest radiographs, and she died 2 weeks after admission. At autopsy, the lung specimen revealed numerous cystic spaces surrounded by a proliferation of smooth muscle cells. Immunohistological examination showed these cells to be positive for HMB-45. Signet cells were seen in the lymphatics and vessels, and hemosiderin-laden macrophages were found in the alveolar spaces. This was a rare case of lymphangioleiomyomatosis with carcinomatous lymphangiosis.

Life style and urinary 8-hydroxydeoxyguanosine, a marker of oxidative dna damage: effects of exercise, working conditions, meat intake, body mass index, and smoking.

The urinary levels of 8-hydroxydeoxyguanosine (8-OH-dG), a marker of oxidative DNA damage, of 318 healthy men aged 18 - 58 were measured with high resolution by a newly developed automated high-pressure liquid chromatography (HPLC) system coupled to an electrochemical detector (ECD). The mean 8-OH-dG level (mg / g creatinine) was 4.12 +/- 1.73 (SD). An eleven-fold inter-individual variation was observed. The accuracy of the measurement estimated from the recovery of an added 8-OH-dG standard was 90 - 98%. By univariate analysis, it was found that moderate physical exercise (P = 0.0023) and high body mass index (BMI) (P = 0.0032) reduced the 8-OH-dG level, while physical labor (P = 0.0097), smoking (P = 0.032), and low meat intake (less than once / week) (P = 0.041) increased its level. Based on a multi-regression analysis of the log-transformed values, moderate physical exercise (P = 0.0039), high BMI (P = 0.0099), and age (P = 0.021) showed significant reducing effects on the 8-OH-dG level, while low meat intake (P = 0.010), smoking (P = 0.013), and day-night shift work (P = 0.044) increased its level. These results suggest that many types of life-style factors that either generate or scavenge oxygen radicals may affect the level of oxidative DNA damage of each individual.

[Relationship between dietary life style in youth and osteoporosis].

The relationship between the dietary life style and osteoporosis was examined by comparing 71 osteoporotic women [age--65.5 +/- 8.9 (mean +/- SD)] with 76 women age-matched as controls. All subjects lived in and around Tsukuba City. Bone mineral density (BMD) was measured by dual energy X-ray absorptiometry. The frequencies of drinking milk and eating meat, fish, or potatoes, etc. both in their youth (about 18-25 years old) and at the present were investigated. The results were as follows: 1) The frequency of having unbalanced diets in their youth was significantly higher in osteoporotic patients than in controls. 2) During their youth, osteoporotic patients had significantly lower frequency of drinking milk, and significantly higher frequency of eating meat, dried fish or eggs, compared with controls. 3) High milk-consumers during their youth (milk-drinking: > or = 3 times per week) were significantly less frequent in osteoporotic patients than that in controls. 4) In the controls, the frequencies of drinking milk and eating meat during their youth were significantly positively correlated with lumbar BMD. The frequency of eating potatoes was significantly negatively correlated with the BMD in controls. 5) No strong relationships between present dietary life and osteoporosis or BMD were found. These findings suggest that drinking milk in their youth may influence BMD and associate with osteoporosis in women.

Simultaneous determination of binding of a variety of radioligands related to ionotropic excitatory amino acid receptors in fetal and neonatal rat brains.

Expression of ionotropic excitatory amino acid receptors was assessed by membrane binding assays using a variety of radioligands in fetal and neonatal rat brains. In fetal rat brain, receptors sensitive to N-methyl-D-aspartate (NMDA) exhibited delayed onset of expression during the last 7 days before birth as compared with those insensitive to NMDA. In addition, developmental increases in agonist-preferring sites preceded those in antagonist-preferring sites within the first 7 postnatal days in particular brain structures with respect to each domain on the NMDA receptor complex. Growth of animals led to drastic increments of [3H](+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d] cyclohepten-5,10-imine (MK-801) binding to the NMDA channel in telencephalic regions until 21 to 28 days after birth, with concomitant desensitization to inhibition by protons of [3H]MK-801 binding in cortical membranes. By contrast, three different agonists were invariably effective in more potently potentiating [3H]MK-801 binding in cortical membranes of 14- and 28-day-old rats than in those of 5-day-old rats. These results suggest that the NMDA-sensitive subclass may play more critical roles in mechanisms underlying postnatal development of rat telencephalon than do the NMDA-insensitive subclasses.

Drug-induced down-regulation of topoisomerase I in human epidermoid cancer cells resistant to saintopin and camptothecins.

The anticancer agent saintopin induces DNA cleavage mediated by both topoisomerase (topo) I and topo II in vitro through stabilization of the reversible enzyme-DNA cleavable complex. We established saintopin-resistant cell lines (KB/STP-1 and KB/STP-2) from human epidermoid cancer KB cells by stepwise exposure to increasing doses of the drug. KB/STP-1 and KB/STP-2 cells showed 12- and 44-fold increases, respectively, in resistance to saintopin relative to that of KB cells. Both saintopin-resistant cell lines showed only small reductions in sensitivity to the topo II inhibitor etoposide but developed marked cross-resistance to the topo I-targeting camptothecin derivative CPT-11 [(4s)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbony loxy] dione hydrochloride trihydrate] and its active form, SN-38 (7-ethyl-10-hydroxycamptothecin). In contrast, both KB/STP-1 and KB/STP-2 cells showed increased collateral sensitivity to cisplatin, a nitrosourea derivative, mitomycin C, and UV light. The protein concentration, activity, and mRNA abundance of both topo I and topo II were similar in KB/STP-1, KB/STP-2, and the parental KB cells. There were no significant changes in the drug-stabilized topo-DNA cleavable complex formation in KB and KB/STP-2 cells. Two point mutations were detected in topo I cDNA from KB/STP-2 cells, but these were also present in KB cells. Topo I mRNA abundance decreased markedly immediately after exposure of KB/STP-2 cells to saintopin; no such effects were apparent in KB cells. In contrast, topo II mRNA was not markedly affected by saintopin in either KB or KB/STP-2 cells. Treatment with CPT-11 or SN-38 also induced a markedly greater and more persistent reduction in topo I mRNA abundance in KB/STP-2 cells than in KB cells. Etoposide had no marked effect on topo I mRNA abundance in either KB/STP-2 or KB cells. Topo I mRNA was highly unstable in KB/STP-2 cells in comparison to KB cells when incubated with saintopin. This novel regulation of topo I mRNA by topo I-targeting agents could be associated with acquirement of drug resistance to saintopin or SN-38/CPT-11 in KB/STP-2 cells.

Decreased DNA topoisomerase II alpha expression and cold-sensitive growth in a mouse mammary cancer cell line resistant to etoposide and doxorubicin.

A mouse mammary carcinoma FM3A cell line resistant to the DNA topoisomerase (topo) II-targeting agent, etoposide (VP-16), FM3A/VP-2B, had a markedly reduced growth rate at a low temperature (33 degrees C). The cells had the following properties: (a) FM3A/VP-2B, which had 24-fold higher resistance to VP-16 than its parental line, FM3A, was cross-resistant to doxorubicin, but not to a camptothecin derivative, CPT-11. (b) Cold-resistant revertants from FM3A/VP-2B, R-6 and R-11, remained 8- to 9-fold more resistant to VP-16 and 2- to 3-fold more resistant to doxorubicin. (c) FM3A/VP-2B had one-fourth the level of topo II activity and one-third of the topo II alpha content and mRNA of FM3A. R-6 and R-11, however, had levels similar to FM3A. (d) FM3A/VP-2B and FM3A had a 3-base deletion at position 4170 on one allele on the topo II alpha cDNA, but expression of the wild-type and the deletion allele was not appreciably changed in both cell lines. Decreased topo II alpha expression might have led to the acquisition of drug resistance to etoposide in FM3A/VP-2B, and appeared to be linked with the cold-sensitive growth. We also present a corrected mouse topo II alpha cDNA sequence.

DNA topoisomerase II alpha gene expression under transcriptional control in etoposide/teniposide-resistant human cancer cells.

We previously isolated etoposide/teniposide-resistant cell lines from human cancer KB cells, designated KB/VP-2 and KB/VM-4, respectively, and we found that decreased expression of topoisomerase II alpha was associated with the acquisition of etoposide/teniposide resistance in both resistant cell lines. In this study, we studied how the expression of the DNA topoisomerase II alpha gene is regulated in drug-resistant cell lines at the transcriptional level. We first examined whether the decreased topoisomerase II alpha mRNA level was due to a shorter lifetime of mRNA molecules in drug-resistant cell lines. A comparison of the degradation kinetics of topoisomerase II alpha mRNA demonstrated that there was no difference in mRNA stability between both resistant cell lines and their parental counterpart. A run-on experiment with isolated nuclei showed that the transcriptional activity of topoisomerase II alpha gene of both resistant cell lines constituted less than 20% of the parental KB cells. The activity of DNA topoisomerase II alpha promoter in resistant cells was also less than 20% of that in KB cells when transient transfection assays were performed with the promoter-driven bacterial chloramphenicol acetyltransferase gene. Among the several transcription factors that might be involved in DNA topoisomerase II alpha gene expression, expression of Sp3, an inhibitory member of the Sp1 family, was elevated to about 3-fold higher in both resistant cell lines than their parental counterpart. These results indicated that the expression of DNA topoisomerase II alpha gene decreased at the transcriptional level through the enhanced expression of Sp3 in our two etoposide/teniposide-resistant cell lines.

Change of retinal correspondence in monocularly reduced intensity of stimulus.

We analyzed the retinal correspondence with a phase difference haploscope on reducing the stimulus intensity of a monocular image with a series of neutral density filters. Ninety-one exotropes were examined by this method. Five cases changed from normal to anomalous correspondence when the stimulation of the strabismic eye was reduced. Two cases showed anomalous correspondence without a filter, but normal correspondence with a reduced stimulus of the normal eye. All these seven cases showed normal correspondence with any other test. The results suggest that masked anomalous correspondence becomes manifest in some cases when the strabismic eye is stimulated more intensely than the normal eye. These cases display both normal and anomalous correspondence and this condition may be called dual correspondence.

[Study of retinal correspondence in esotropia with phase difference haploscope].

There is some concern regarding the evaluated depth of anomalous correspondence, because we often experience paradoxical results. We have already reported that, in intermittent exotropia, we sometimes found dual correspondence with a phase difference haploscope (PDH) reducing the stimulus intensity of either image with a series of neutral density filters. In this study, we analyzed the retinal correspondence in esotropes with the same procedure. 27 esotropes were examined. 11 cases had normal correspondence and 16 cases anomalous correspondence in PDH. All of the 11 normal correspondence cases retained normal correspondence when we reduced the stimulus intensity of either image of fixating eyes or squinting eyes. In the 16 anomalous correspondence cases in PDH, 2 cases showed normal correspondence when the stimulus intensity of the fixating eyes was reduced. On the other hand, 3 cases showed normal correspondence when the stimulus intensity of the squinting eyes was reduced. In fixed heterotropia, both the fovea and the part of the retina of the squinting eye which received the same images as the fovea of the fixating eye (fixating point), were suppressed by the fixating eye. We consider that, in some cases with anomalous retinal correspondence in PDH, when the stimulus intensity of the fixating eyes or the squinting eyes is reduced, the balance of the sensitivity of the foveas and the fixating points of the squinting eyes change, and show normal correspondence.

Preconcentration of traces of silver and bismuth from cobalt and nickel and their nitrates, with activated carbon.

A method is described for the preconcentration of trace metals Ag and Bi, present as impurities in high-purity cobalt and nickel metals and their nitrates. After the metal samples have been dissolved in nitric acid (or the salts in water) the trace elements are complexed with ammonium pyrrolidinedithiocarbamate (APDC). The sample solution is then filtered through a 2-cm filter paper coated with 50 mg of activated carbon, whereby the complexed trace metals are adsorbed on the activated carbon and separated from the matrix. The trace elements are dissolved off with nitric acid and determined by flame atomic-absorption spectrometry (AAS). The detection limits for the analysis of 10 g of metal samples and 50 g of the nitrate samples were 0.002-0.035 ppm Ag and 0.04 ppm Bi for the metal samples, and 0.0003-0.0004 ppm Ag and 0.004-0.005 ppm Bi for the nitrate samples. The coefficient of variation, in general, is 10-25% for Ag and 33% for Bi.