Association between gut microbiota, bone metabolism, and fracture risk in postmenopausal Japanese women.

INTRODUCTION: We investigated the relationship between gut microbiota composition and osteoporosis/fracture risk in Japanese postmenopausal women using 16S rRNA gene sequencing, FRAX, bone mineral density, biochemical bone parameters, and a self-administered questionnaire. Variation in abundance of specific microbiota was found to be significantly associated with fracture risk and vitamin K levels. Gut microbiota data with respect to bone metabolism and fracture risk is limited. Vitamin K is produced by certain intestinal bacteria and has been reported to play a role in maintaining bone quality. PURPOSE: We investigated relationships among gut microbiota composition, bone metabolism, and fracture risk in postmenopausal Japanese women. METHODS: Bone mineral density (BMD) was evaluated in 38 postmenopausal women (mean age 62.9 years) using forearm dual-energy X-ray absorptiometry. We collected and analyzed serum bone turnover markers (vitamin K fraction and tartrate-resistant acid phosphatase 5b; TRACP-5b), gut microbiota profiling (16S rRNA gene sequencing), and self-administered questionnaire data, including fracture history and vitamin K intake. Vitamin K2, BMD, and TRACP-5b data were divided into high- and low-level groups using cutoff values of 0.06 ng/mL, 87.05%, and 420 mU/dL, respectively; the proportions of bacteria were analyzed. Fracture incidence and relative risk were investigated for each bacterium. RESULTS: The genus Bacteroides was predominant in the high vitamin K2 group (29.73% vs 21.58%, P = 0.022). Fracture incidence was significantly higher in the low Bacteroides group, with a 5.6-times higher risk ratio of fracture history. The family Rikenellaceae was more abundant in the low BMD group and more abundant in the high TRACP-5b group (2.15% vs 0.82%, P = 0.004; 2.38% vs 1.12%, P = 0.013, respectively). CONCLUSION: Bacteroides and Rikenellaceae may be involved in bone metabolism and fracture risk. Further investigations of the underlying microbiota-related pathways in bone metabolism may reveal treatment strategies, and facilitate the prevention of osteoporosis.

Edaravone (MCI-186) is effective as a free radical scavenger following arteriovenous sheathotomy for treatment of macular oedema associated with branch retinal vein occlusion.

AIMS: To determine whether edaravone (MCI-186), a free radical scavenger, can reduce macular oedema and improve the visual acuity after arteriovenous sheathotomy in eyes with a branch retinal vein occlusion (BRVO). METHODS: Forty-seven eyes of 47 consecutive patients with a BRVO who were treated with arteriovenous sheathotomy were studied. The patients were assigned prospectively to either Group R who received 30 mg of edaravone (Radicut) systemically during the vitrectomy or Group N who did not receive any drugs. The postoperative visual acuity was measured before and 12 months after the operation. RESULTS: At 12 months postoperatively, the best-corrected visual acuity (BCVA) in logarithm of the minimum angle of resolution (logMAR) units improved significantly from 0.22 to 0.56 logMAR units in Group R and from 0.20 to 0.27 units in Group N (p = 0.016). Twenty-three of 27 cases (85%) in Group R and four of 15 cases (27%) in Group N showed an improvement in BCVA of >0.2 logMAR units (p = 0.0025). CONCLUSION: The better visual acuity in patients given edaravone than those without endaravone during the arteriovenous sheathotomy suggests that edaravone improved the physiology of the retinal cells after the arteriovenous sheathotomy.

Search for cosmic-ray antideuterons.

We performed a search for cosmic-ray antideuterons using data collected during four BESS balloon flights from 1997 to 2000. No candidate was found. We derived, for the first time, an upper limit of 1.9 x 10(-4) (m2s sr GeV/nucleon)(-1) for the differential flux of cosmic-ray antideuterons, at the 95% confidence level, between 0.17 and 1.15 GeV/nucleon at the top of the atmosphere.

Impact of reproductive experience on women's smoking behaviour in Japanese nurses.

OBJECTIVES: The objectives of this study were to describe current and past smoking behaviour of female Japanese nurses, to examine factors associated with developing and stopping a smoking habit, and to examine how their reproductive experiences affect their smoking behaviour. STUDY DESIGN: A cross-sectional study. This study was a baseline survey of a prospective occupational cohort study. METHODS: A self-administered survey of 1748 female Japanese nurses aged over 20 years was conducted in Gunma prefecture, Japan, in 1999. RESULTS: Overall, 27.2% developed a smoking habit (current smokers, 19.8%; ex-smokers, 7.4%) and 72.2% were never smokers. Logistic regression analysis showed that the type of nursing certificate was statistically associated with developing a smoking habit. It also showed that the type of nursing certificate, work place, marital status and current pregnancy were statistically associated with smoking cessation. Currently pregnant women were more likely to stop smoking than non-pregnant women (adjusted odds ratio, 3.18; 95% confidence intervals, 1.25-8.06). For women aged 20-29 years, the proportions of current smokers, ex-smokers and never smokers among pregnant women were 11.5, 23.1 and 65.4%, respectively; among non-pregnant women of this age, the values were 22.3, 4.3 and 73.4%, respectively. There was a statistically significant difference in such proportions between the two groups (chi2=19.27; P<0.0001). More than half of the ex-smokers who were currently pregnant had stopped smoking in the last 12 months. Smoking behaviour showed no statistically significant difference between women who had had at least one delivery and women who had not. CONCLUSIONS: The results suggest that pregnancy provides a good opportunity for smoking cessation, but a large proportion of women who successfully quit smoking during pregnancy relapse after delivery.

Microstructures formed on a low fluorescent glass using glassy carbon molding.

We report a pattern generation method on galss using hot embossing technology. Microstructures were formed on Borofloat, a low fluorescent glass, using a glassy carbon mold. A cubic block in the size of 100 um x 100 um x 50 um is the basic testing pattern. This block was repeated on a mirror polished glassy carbon wafer with a space in the pitch of 30 um. The whole pattern area is in 15 mm x 15 mm square on this carbon mold. Both the glass wafer and the mold were heated to 655 °C and embossing was processed at 2 MPa under vacuum (0.07 Pa). The state was held for 20 min, and then the embossed piece was cooling down to 200 °C naturally in the vacuum chamber. The pattern on mold were transferred compensate to the glass. Each fabrication cycle is about 1.5 hours. This method shows great potential to fabricate bio-MEMS devices efficiently with a very low cost. The main target is to fabricate the devices for highly sensitive fluorescent detection applications, which is very difficult to be realized using plastic substrates, A multi-channel pattern (with 70 um line and 400 um space) has been generated on a 20 mm square glassy carbon using a laser machine. The pattern replicated on glass chips has been demonstrated.

Detection of tryptase-, chymase+ cells in human CD34 bone marrow progenitors.

BACKGROUND: Mast cells (MCs) arise from haematopoietic stem cells. We have recently reported that CD34(+) progenitors derived from human bone marrow (BM) develop into tryptase+, chymase+ MCs when cultured in the presence of recombinant human stem cell factor (rhSCF) and recombinant human IL-6 (rhIL-6). In an experiment for the expression of chymase during differentiation, chymase+ cells were detected in human BM, but tryptase+ cells were not found. OBJECTIVE: The purpose of this study was to show the appearance of chymase+ cells in CD34(+) cells with an origin different from MC differentiation. METHODS: CD34(+) cells from human BM were sorted with anti-CD117 monoclonal antibody (mAb), and cytospins of CD34(+), CD34(+)CD117(+), or CD34(+)CD117(-) were prepared. These cells were cultured with rhSCF+rhIL-6 for 12 weeks. Some of the cells were subjected to either histological stain with Wright-Giemsa or immunocytochemistry with anti-chymase mAb. Real-time RT-PCR was also performed to compare the transcriptional level of chymase from each cell preparation. RESULTS: Chymase was expressed in CD34(+) cells as well as human MCs by immunocytochemistry. Substantial CD34(+)CD117(-) cells, but not CD34(+)CD117(+) cells, were stained immunocytochemically with anti-chymase mAb. For 1 week culture with rhSCF+rhIL-6, no cells expressed chymase in any preparation. Real-time RT-PCR revealed positivity for chymase mRNA in CD34(+) cells, but it reduced at 1 week of culture, and increased as cells developed into MCs. Chymase mRNA in CD34(+)CD117(+) cells was negligible compared with that in CD34(+)CD117(-). Tryptase mRNA was below the detectable level in CD34(+) cells, and increased along with MC differentiation. After 12 weeks of culture, CD34(+)CD117(+) developed predominantly into MCs, whereas CD34(+)CD117(-) developed into monocytes/macrophages. CONCLUSION: Our findings suggested that chymase is present not only in MCs but also in CD34(+)CD117(-) BM progenitors, but that its origin is different from the MC lineage.

Functional expression of high-affinity receptor for immunoglobulin E on mast cells precedes that of tryptase during differentiation from human bone marrow-derived CD34 progenitors cultured in the presence of stem cell factor and interleukin-6.

BACKGROUND: CD34(+) progenitor cells develop into tryptase(+), CD117(+) mast cells when cultured in the presence of recombinant human stem cell factor (rhSCF). However, spontaneous IgE receptor (FcepsilonRI) expression during human mast cell development is not well examined. OBJECTIVE: Here, the expression and function of FcepsilonRI in and on human bone marrow-derived mast cells (HBMMCs) during development were investigated. METHODS AND RESULTS: At 4 weeks of culture, predominant cells expressed high-affinity IgE receptor alpha chain (FcepsilonRIalpha) on the cell surface determined by flow cytometry, but CD117 was less expressed. Immunocytochemistry with antitryptase mAb and anti-FcepsilonRIalpha mAb revealed intracellular and surface expression of FcepsilonRIalpha at 2 weeks of culture, but tryptase was less expressed. FcepsilonRIalpha mRNA transcript preceded that of tryptase mRNA at 2 weeks of culture determined by real-time RT-PCR, and FcepsilonRIalpha, FcepsilonRIbeta, FcepsilonRIgamma, and tryptase mRNA increased along with differentiation. FcepsilonRIalpha cross-link on HBMMC and 4-week-old mast cells/mast cell precursors induced the release of IL-5 and granulocyte macrophage-colony stimulating factor, which was enhanced by rhSCF. CONCLUSION: These data indicated that HBMMC constitutively and spontaneously expressed functional FcepsilonRI subunits at the early stage of differentiation, probably because of the differences in the ability and functional property of progenitors.

An angiogenesis inhibitor TNP-470 (AGM-1470) suppresses vascular smooth muscle cell proliferation in experimental rat aortotomy models.

AIM: Vascular intimal hyperplasia is an important clinical concern in vascular diseases, such as anastomotic stricture as a possible complication of cardiovascular surgery. We recently suggested that a rat aortotomy model could be substituted for a vascular anastomotic stricture around a suture line. TNP-470 is known as an angiogenesis inhibitor and has demonstrated abilities to inhibit DNA synthesis of smooth muscle cells (SMCs) and SMCs proliferation. The aim of this study was to investigate the effect of TNP-470 on SMC proliferation using rat aortotomy models. METHODS: Longitudinal aortotomy was performed in the abdominal aorta of rats. Rats received a subcutaneous injection of materials (TNP-470, 20 mg/kg) or vehicle 3 times a week (n=10 in each group). The aorta was harvested 2 weeks after aortotomy. Serial sections from tissues were stained with hematoxylin and eosin, and the ratio of intimal to medial cross-sectional areas (I/M ratio) was determined. Values are expressed as the mean +/- the standard deviation. Results. Thickening of the intimal layer 2 weeks following aortotomy was observed in the control group however, intimal thickening was inhibited in the TNP-treated group. The I/M ratio was significantly (p = 0.0376) lower in the TNP-treated group than in the control group (8.3 +/- 4.8 vs 15.6 +/- 9.6%). Conclusion. TNP-470 significantly suppressed intimal thickening in experimental rat aortotomy models. TNP-470 might inhibit the development of anastomotic stricture after cardiovascular surgery.

Survival of allografted pancreatic islets in the subretinal space in rats.

The purpose of this study was to determine whether the subretinal space would provide immune privilege for pancreatic islet transplantation. Islets from outbred Sprague Dawley (SD) rats were isolated by collagenase digestion, and about 200 handpicked islets were transplanted into the subretinal space of SD rats. Similar grafts were transplanted into the subconjunctival space of SD rats as controls. Transplanted eyes were enucleated after 2 to 60 days, fixed and embedded in paraffin for immunoperoxidase staining of insulin, glucagon, and CD8+ lymphocytes. Clinical examination of rat eyes revealed minimal or no inflammation in the anterior chamber or vitreous at any time point. Fifteen of 19 subretinal allografts survived up to 60 days. Few CD8+ lymphocytes were present in the subretinal grafts and the endocrine cells stained intensely for insulin and glucagon at all time points. In contrast, CD8+ lymphocytes were present in subconjunctival grafts in rats by day 14 and all grafts were destroyed by day 21. These results suggest that the subretinal space provides immune privilege for islet allotransplantation by preventing massive lymphocyte infiltration.

Measurements of cosmic-ray low-energy antiproton and proton spectra in a transient period of solar field reversal.

The energy spectra of cosmic-ray low-energy antiprotons ( *p's) and protons ( p's) have been measured by BESS in 1999 and 2000, during a period covering reversal at the solar magnetic field. Based on these measurements, a sudden increase of the *p/p flux ratio following the solar magnetic field reversal was observed, and it generally agrees with a drift model of the solar modulation.

[Antimicrobial susceptibility pattern and epidemiological investigation of the clinical isolates of Pseudomonas aeruginosa in 1999and 2000].

During 1999-2000, a total of 1,215 isolates of Pseudomonas aeruginosa (P. aeruginosa) were analyzed by wards, type of clinical specimens and serotypes. Antimicrobial susceptibility was also examined for 10 agents (PIPC, CAZ, LMOX, IPM-CS, GM, AMK, MINO, LVFX, NFLX, and ST) with criteria by NCCLS (M7-A5). Sputum (33%) was most common source of P. aeruginosa followed by urine (17) and wound discharge (14%). According to wards, the isolation rate at the critical care unit was the highest (40%) in 1999 but decreased to 14% in 2000. In the isolates from inpatients (1999, 2000) the resistance rate was 96.8-98.9% for MINO and ST, 23.5-30.8% for IPM-CS, 27.2-28.9% for LMOX, 12.5-20.1% for GM, and 6.7-11.8% for NFLX. Comparison between the two years showed increases in isolates resistant to GM (p<0.01) and NFLX (p<0.05) but a decrease in IPM-CS resistant isolates (p<0.01). According to serotypes, the E and G type were most frequently observed (242 isolates each, 21.8%) followed in order by F type (127 isolates, 11%) and the B type (124 isolates, 11%). The incidence of the F type was high compared with other medical institutions. There was a reduction in the rates of susceptibility of serotype E isolates to PIPC (74.4%) and GM (60.3%), and of serotype F isolates to IPM-CS (52%). Multidrug-resistant P. aeruginosa (MDR-PSA) that were resistant to beta-lactams, aminoglycosides, and new quinolons accounted for 3%.

Pulmonary intravascular lymphoma complicated with Pneumocystis carinii pneumonia: a case report.

We report a case of intravascular lymphoma (IVL) complicated with Pneumocystis carinii pneumonia (PCP). A 65-year-old male complaining of dyspnea and dementia was diagnosed to have pulmonary IVL by transbronchial lung biopsy. Concomitantly, deoxyribonucleic acid sequence specific to Pneumocystis carinii was detected in bronchoalveolar lavage fluid by polymerase chain reaction. Differential responses to the sequential treatments for PCP and IVL implied that increased serum lactate dehydrogenase (LDH) was due to PCP, whereas hypoxemia and dementia were due to IVL. Although pulmonary IVL and PCP share many clinical presentations, exact diagnosis is essential for their successful treatment.

Indomethacin induced bulky lymphadenopathy and eosinophilic pneumonia.

Indomethacin is one of the most popular non-steroidal anti-inflammatory drugs (NSAID). Although NSAID occasionally provoke bronchospasm and hypersensitivity pneumonia, they seldom cause lymphadenopathy. This is the first report in which NSAID induced both eosinophilic pneumonia and bulky intrathoracic lymphadenopathy simultaneously. A 76-year-old Japanese man experienced high fever and dyspnoea after using an indomethacin suppository. Computed tomography scan of his chest revealed massive mediastinal and hilar lymphadenopathy along with diffuse infiltration in both lungs. He was diagnosed to have eosinophilic pneumonia because of eosinophilia in his peripheral blood and bronchoalveolar lavage fluid (BALF). Without using glucocorticoids, the pulmonary infiltration and lymphadenopathy subsided spontaneously. As the blastoid transformation test using the lymphocytes in his BALF was positive to indomethacin, we judged that both his eosinophilic pneumonia and mediastinal lymphadenopathy were due to a hypersensitivity reaction to indomethacin. An allergic reaction to NSAID should be considered as a rare cause of mediastinal lymphadenopathy.

Invasive pulmonary aspergillosis in a puerperant with drug-induced agranulocytosis.

Invasive pulmonary aspergillosis (IPA) is an acute infection of Aspergillus species to the lungs. It generally occurs in immunocompromised hosts, especially with neutropenia. We report a 30-year-old puerperant, who developed IPA from agranulocytosis. She had been treated for threatened labor with ritodrine and cefepime, one of which induced agranulocytosis. After vaginal delivery of twins, pneumonia emerged in the right lower lobe. She was diagnosed to have IPA according to the halo sign on computed tomography (CT) and positive circulating antibody against Aspergillus, and was treated successfully with oral itraconazole followed by surgical resection. It is important to note that IPA might arise in otherwise immunocompetent hosts when neutropenia is long-standing.

[PL granule-induced pneumonia requiring mechanical ventilation].

PL granule is one of the most common forms of composite medicine for colds in Japan, including acetaminophen. We report a case of PL granule-induced pneumonia that required mechanical ventilation. A 72-year old man who had been prescribed amiodarone for more than one year before for arrhythmogenic right ventricular dystrophy, repeatedly took PL granules at one-week intervals because of sneezing and fever. He then underwent acute respiratory failure. He needed mechanical ventilation for severe hypoxemia, but recovered with glucocorticoid pulse therapy. Because the blastoid transformation test using his peripheral blood lymphocytes was positive on stimulation with PL granules, but negative on amiodarone stimulation, he was diagnosed as having drug (PL granule)-induced pneumonia. Only one case of PL granule-induced pneumonia and seven of acetaminophen-induced pneumonia have been reported. None of these patients needed mechanical ventilation. PL granule-induced pneumonia should be treated cautiously because of possible acute exacerbation.

Precision measurement of cosmic-Ray antiproton spectrum

The energy spectrum of cosmic-ray antiprotons ( &pmacr;'s) has been measured in the range 0.18-3.56 GeV, based on 458 &pmacr;'s collected by BESS in a recent solar-minimum period. We have detected for the first time a characteristic peak at 2 GeV of &pmacr;'s originating from cosmic-ray interactions with the interstellar gas. The peak spectrum is reproduced by theoretical calculations, implying that the propagation models are basically correct and that different cosmic-ray species undergo a universal propagation. Future BESS data with still higher statistics will allow us to study the solar modulation and the propagation in detail and to search for primary &pmacr; components.

Argatroban inhibits intraocular fibrin formation after vitrectomy in rabbits.

OBJECTIVES: To examine whether the specific thrombin inhibitor argatroban can prevent anterior chamber, pupillary area, and anterior vitreous fibrin formation after vitrectomy and lensectomy in rabbits. METHODS: Argatroban was infused into the vitreous cavities of Japanese albino rabbits for 5 minutes after pars plana vitrectomy and lensectomy. Slitlamp microscopy and indirect ophthalmoscopy were performed at postoperative hours 0.5, 1, 2, 3, and 6, and at postoperative days 1, 2, 3, and 7, and the amounts of fibrin formation in the anterior chamber, pupillary area, and anterior vitreous were scored from grade 0 to 4. RESULTS: Argatroban prevented fibrin formation from 0.5 hours postoperatively in a dose-dependent manner. In the eyes treated with 0.01% argatroban, the median score for postoperative fibrin formation was significantly less than that in control eyes between hours 1 and 3 (hour 1, P =.02; hour 2, P =.005; and hour 3, P =.003); the eyes treated with 0.003% argatroban also had significantly less fibrin than control eyes between 1 and 2 hours (hour 1, P =.005; hour 2, P =.03). CONCLUSION: These results indicate that argatroban inhibits intraocular fibrin formation in an experimental rabbit model. CLINICAL RELEVANCE: Argatroban may be useful clinically in cases that often produce fibrin postoperatively, such as proliferative vitreoretinopathy and proliferative diabetic retinopathy.

Pleural amyloidosis in a patient with intractable pleural effusion and multiple myeloma.

Pleural involvement of systemic amyloidosis has been rarely reported. We report a case with multiple myeloma presenting an intractable right pleural effusion, in which pleural amyloidosis was diagnosed through pleural biopsy using a Cope needle. The diagnosis of pleural amyloidosis is important, because its refractory pleural effusion should be treated with pleurodesis. Since closed pleural biopsy using a Cope needle is much less invasive than thoracoscopy, the former should be attempted first whenever pleural amyloidosis is suspected.

Nuclear hypersegmentation precedes the increase in blood eosinophils in acute eosinophilic pneumonia.

In this case of acute eosinophilic pneumonia (AEP), eosinophils with hypersegmented nuclei emerged in the blood before the increase of eosinophil count. An 18-year-old woman complaining of fever, cough and dyspnea was admitted because of diffuse ground-glass opacities in her chest roentgenogram. On admission, her blood cell count revealed a marked increase of neutrophils. Although the number of eosinophils was normal, some of them contained three- or four-lobed nuclei. She was diagnosed to have AEP through bronchoalveolar lavage and transbronchial lung biopsy. The combination with acute clinical course, pulmonary infiltration and the presence of hypersegmented eosinophils in blood may imply the diagnosis of AEP.