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We conducted a retrospective study among 497 male patients with OSA on CPAP therapy. Participants with pretreatment Apnea-Hypopnea Index (AHI) data based on overnight polysomnographic recordings completed a questionnaire. Adherence data for CPAP therapy were collected using a smart card system. We classified CPAP use of ≥4 hours per night and ≥70% of nights as good adherence; other CPAP use was categorized as poor adherence. Logistic regression models were used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for poor adherence to CPAP therapy in the hypertension and diabetes mellitus groups, compared with the no comorbidity group, adjusting for body mass index, duration of CPAP therapy, AHI, and Epworth Sleepiness Scale score.In the no comorbidity, hypertension, and diabetes mellitus groups, 43.4%, 44.7%, and 56.0%, respectively, had poor adherence to CPAP therapy. Being in the diabetes mellitus group was significantly associated with poor adherence to CPAP therapy (OR=1.86, 95% CI: 1.18-2.92,Our results indicate that comorbidity of diabetes mellitus is associated with poor adherence to CPAP therapy in male patients with OSA.

To investigate the contributions of adenoid and tonsil sizes to obstructive sleep apnea syndrome (OSAS) in normal-weight children in two age categories: preschool and schoolchildren.Fifty-eight normal-weight (body mass index z-score2) symptomatic children with OSAS (apnea-hypopnea index ≥ 2) were evaluated. The patients were divided into two age categories: preschool (age6; n=33) and schoolchildren (age ≥ 6; n=25). Polysomnographic findings and adenoid and tonsil sizes were compared. The relative contributions of body mass index and adenoid and tonsil sizes were also investigated with a regression analysis.Adenoid grade and apnea index correlated significantly in preschool children (r=0.45, p0.01). On regression analysis, adenoid grade was a significant predictor of the apnea index in preschool children. The influence of adenoid hypertrophy decreased from preschool to schoolchildren. Tonsil size had little influence on the apnea index in either group.Adenoid hypertrophy was a major contributor to OSAS in normal-weight preschool children. The upper airway morphology of younger children with OSAS differed from that of older children with OSAS.

Ayako Saito,1 Shigeko Kojima,2 Fumihiko Sasaki,3 Masamichi Hayashi,4 Yuki Mieno,4 Hiroki Sakakibara,5 Shuji Hashimoto1 1Department of Hygiene, School of Medicine, Fujita Health University, Toyoake, Japan; 2Department of Rehabilitation, Faculty of Health Sciences, Nihon Fukushi University, Handa, Japan; 3SDB Research Laboratory, Takaoka Clinic, Nagoya, Japan; 4Department of Internal Medicine, Division of Respiratory Medicine and Clinical Allergy, Fujita Health University, Toyoake, Japan; 5Tokushige Kokyuki Clinic, Nagoya, Japan Abstract: The purpose of this study was to develop and evaluate a self-efficacy instrument for Japanese obstructive sleep apnea (OSA) patients treated with continuous positive airway pressure (CPAP). Analyzed subjects were 653 Japanese OSA patients (619 males and 34 females) treated with CPAP at a sleep laboratory in a respiratory clinic in a Japanese city. Based on Bandura's social cognitive theory, the CPAP Self-Efficacy Questionnaire for Sleep Apnea in Japanese (CSESA-J) was developed by a focus group of experts, using a group interview of OSA patients for the items of two previous self-efficacy scales for Western sleep apnea patients receiving CPAP treatment. CSESA-J has two subscales, one for self-efficacy and the other for outcome expectancy, and consists of a total of 15 items. Content validity was confirmed by the focus group. Confirmatory factor analysis showed that the factor loadings of self-efficacy and outcome expectancy were 0.47–0.76 and 0.41–0.92, respectively, for the corresponding items. CSESA-J had a significant but weak positive association with the General Self-Efficacy Scale, and a strong positive association with “Self-efficacy scale on health behavior in patients with chronic disease.” Cronbach’s alpha coefficient was 0.85 for the self-efficacy subscale and 0.89 for the outcome expectancy subscale. The intraclass correlation coefficient using data from the first and second measurements with CSESA-J for a subset of 130 subjects was 0.93 for the self-efficacy and outcome expectancy subscales. These results support CSESA-J as a reliable and valid instrument for measuring the self-efficacy of Japanese OSA patients treated with CPAP. Further studies are warranted to confirm validity for female OSA patients and generalizability. Keywords: scale development, obstructive sleep apnea, continuous positive airway pressure, social cognitive theory

We studied the change in the hypothalamo-pituitary-adrenal gland (HPA) axis upon adding prior toluene inhalation to our previous formaldehyde inhalation experiments to determine whether short term exposure to relatively high levels of toluene triggers multiple chemical sensitivity (MCS). Data come from immunocytochemical, morphometrical and RT-PCR measurements. Four groups of adult female mice were exposed to differing concentrations (0, 80, 400, and 2,000 ppb) of formaldehyde for 16 hr/day, 5 days/week for twelve weeks, after the mice were exposed intranasally to 500 ppm toluene per mouse for 6 hr/day, for 3 days. We found that the number of corticotropin releasing hormone (CRH)-immunoreactive (ir) neurons was up-regulated according to the amount of formaldehyde as well as inhalation of formaldehyde alone in our previous experiment. The proportion of adrenocorticotropin hormone (ACTH)-ir cells increased according to the formaldehyde concentration, though there was no significant difference between the 400 and 2,000 groups. The number of ACTH-ir cells was higher in the 400 group than in the other groups (0, 80, and 2,000). Expression of ACTH-mRNA was also up-regulated according to the quantity of formaldehyde. The sinusoid in the anterior pituitary showed more dilatation in the 400 and 2,000 groups than in the control group, especially in the 2,000 group. We propose that exposure to toluene prior to inhalation of formaldehyde has no effect on the HPA axis and as a trigger of MCS, although greater sinusoid dilatation was found in the anterior pituitary gland at higher concentrations of formaldehyde.

We have examined alterations in the hypothalamo-pituitary GH-somatic growth axis and the hypothalamo-pituitary LH-ovarian axis in a line of transgenic ICR mice expressing human GH (hGH) under the influence of the whey acid protein promoter. Transgenic female mice weighed twice as much as control females and were infertile. The size of the anterior pituitary (AP) was 1/3 that of the controls. In transgenic mice, acinar cells in the mammary and mandibular glands displayed hGH-immunoreactivity, and plasma hGH was detected by radioimmunoassay. In the medial basal hypothalamus (MBH) of transgenic females, the immunoreactive-GHRH level was decreased (P

Gicerin is a cell adhesion molecule in the immunoglobulin (Ig) superfamily and plays an important role during development through its adhesive properties. Gicerin has two isoforms that differ in their cytoplasmic domains; s-gicerin is the shorter and l-gicerin the longer form of the protein. Gicerin is over-expressed in some sporadic tumors as well as in developing tissues. To provide direct evidence that gicerin has the potential to participate in malignant aspects of tumor cell behavior, a gicerin cDNA was introduced into L-929 cells, an endogenous gicerin-negative mouse fibroblast and subsequently analyzed for changes in their invasive and metastatic potential by implantation into nude mice and chick embryos. Compared with parental cells, both gicerin isoform transfectants showed an enhanced cell growth and invaded deeply into surrounding tissues from implanted sites in both animal models. Furthermore, l-gicerin transfectants markedly enhanced metastasis to the lung. These findings suggest that gicerin promotes the tumor growth and invasion, and the isoform bearing the longer cytoplasmic domain may play a role in metastasis. J. Cell. Physiol. 197: 103–109, 2003© 2003 Wiley-Liss, Inc.

We examined elastic fibrils in the aortic intima of aged rats with elastin staining, elastase digestion and X-ray microanalysis. The innermost elastic lamina, heavily stained by a brief treatment with azure II-toluidine blue without heating, was thin and fragmented, and an amorphous substance that stained weakly to moderately was accumulated in the thick subendothelial space of 26-28-month-old rat aortas. The substance was always present in the intimas; to a large extent in the aged rats but rarely in 5-month-old rats, and disappeared after digestion with elastase. The amorphous substance was identified as elastic fibrils approximately 0.1-0.2 microm in diameter under an electron microscope with tannic acid-uranyl acetate/lead citrate staining. Elastase digestion revealed that elastic fibrils were composed of a number of microfibrils, which were 10-12 nm in diameter, in and around a fine core of amorphous elastin. X-ray microanalysis revealed a clear peak of sulphur in the elastic fibrils.

Mitochondria occasionally increase in size in response to metabolic injury. Numerous studies have reported giant mitochondria in patients with various diseases and animals with metabolic injuries, but there are few reports on giant mitochondria in normal cells under physiological conditions. Here, we report giant mitochondria in normal gastric parietal cells. Stomachs of guinea pigs fed freely, fasted or fasted and then injected with histamine were processed for electron microscopy. Giant mitochondria >2 microm in the diameter of their major axis were observed in resting-type parietal cells in the gastric glands of animals fasted for 60-72 h, whereas acid-secreting-type parietal cells found in those fed ad libitum did not contain giant mitochondria. Giant mitochondria showed unusual structures, especially in their cristae: they contained closely packed, tubular and concentric cristae as well as amorphous and pleomorphic inclusion bodies in their matrix. We observed giant mitochondria consisting of several segments, suggesting the fusion of several normal-sized mitochondria. Histamine injection decreased in a frequency of giant mitochondria in accordance with a decrease in a frequency of resting-type parietal cells. This is the first report of giant mitochondria in gastric parietal cells under physiological or near physiological conditions. Gastric parietal cells might be a good model for examining mitochondrial fusion and fission in a physiological state accompanied by the morphological change of the cells in the membrane system from an acid-secreting to resting type.

Pituitary glands of chicken, from stages 20 (70 ∼ 72 h of incubation) to 46 (20 days) of Hamburger and Hamilton (1951), were studied by immunocytochemical and histological stainings and India ink injection into blood vessels. Using the distribution pattern of 6 types of immunoreactive adenohypophyseal cells and the location of pituitary stalk as guideposts, we found how specific areas in the epithelium of Rathke’s pouch differentiate into specific regions of the adenohypophysis at 20 days. In the sagittal plane, the walls of Rathke’s pouch were tentatively divided into the upper part (A1 + A2) and lower part (A3) of the anterior wall, and the posterior wall (P1 + P2 + P3). The cephalic lobe was mainly assembled by the proliferation of parenchymal cells in the areas A2 + A3 + P2 of Rathke’s pouch epithelia at 3 days of incubation. The caudal lobe was derived from A1 + P1 + P3. The pars tuberalis was derived from A1 + A2. Thus, the avian adenohypophysis is established at 13 days, though the blood supply to the pars distalis is established at 20 days. Therefore, the cephalic lobe and caudal lobe of the pars distalis and the pars tuberalis of the chicken adenohypophysis are derived from specific areas of the cell cords of Rathke’s pouch at 3 days of incubation.

We previously found that parietal cells in guinea pigs starved for a few days contained giant mitochondria while the cells of animals starved and then injected with histamine mostly did not. To test whether mitochondria change their size and activity according to energy demands for acid secretion, we examined the dynamics of mitochondria stained by rhodamine 123 in living parietal cells in gastric glands under a confocal microscope. The glands were isolated from guinea pigs starved for 60-72 hr. Because mitochondria were closely packed in the cytoplasm, we failed to observe the morphological changes of each mitochondrion in parietal cells. However, we successfully observed that 10-5 M histamine induced an increase in the fluorescence intensity (the concentration of rhodamine 123) in mitochondria in subpopulations of parietal cells; the fluorescence intensity increased sharply within minutes after the histamine administration in some cells, while it gradually increased from just after the administration in other cells. We also found that a subpopulation of mitochondria within a parietal cell responded to the secretagogue. The findings suggest that parietal cells exist as a heterogeneous population in gastric glands and contain heterogeneous mitochondria in terms of their mitochondrial response to histamine.

A new method of photochemical generation of sulfonic acid is described. N-sulfonyl-arylsulfonamide which is synthesized easily from arylsulfonamide and sulfony chloride produce sulfonic acids on photoexcitation. Quantum yields of their decomposition are between 0.2 to 0.5, depending on their substituents. A typical example is N-(4-methoxyphenysulfony)-N-methyl-4-methoxyphenysulfonamide which shows quantum yield of 0.5. It is almost the same value as commonly used photo-sulfonic acid generating compounds. Photodecompositon products were characterized by HLPC and Mass spectroscopy, and photodecompositon mechanism is proposed.

In this study we investigated the effect of adrenomedullin (AM) on fMLP-mediated activation of human neutrophils. AM partially, but significantly, suppressed fMLP-induced upregulation of CD11b expression. The inhibitory effects of AM upon fMLP-induced upregulation of CD11b expression were completely blocked by CGRP [8-37], a CGRP receptor antagonist. AM significantly increased cAMP content in neutrophils and SQ-22,536, an adenylate cyclase inhibitor, and KT-5720, a PKA inhibitor, significantly blocked the inhibitory effects of AM upon fMLP-induced upregulation of CD11b expression. This study indicates that binding of AM to the CGRP receptor suppresses fMLP-induced upregulation of CD11b expression of human neutrophils by increasing intracellular cAMP levels. AM may play an important role in the regulation of inflammatory processes, especially in the binding of neutrophils to vascular endothelial cells and subsequent neutrophil emigration evident in acute pulmonary inflammation.

Hypoxia induces bronchodilation in vivo and in vitro, but the mechanisms are still unclear. To evaluate whether an extra- or intracellular free Ca(2+) ion is involved in the mechanisms of hypoxic relaxation, we simultaneously measured cytosolic Ca(2+)levels and tensions in both intact and denuded guinea-pig tracheal strips precontracted with histamine (100 microM), and assessed the effect of hypoxia on guinea-pig tracheal rings precontracted with okadaic acid (10 microM) and calyculin-A (0.1 approximately 10 microM) under an extracellular Ca(2+)-free state. The exposure of tracheal rings to hypoxia induced an immediate decrease of tracheal tension without decrease in intracellular free Ca(2+)levels. In the presence of okadaic acid but not calyculin-A, hypoxic air exposure caused significant transient reductions in tracheal tone. Further, thapsigargin (5 microM or 10 microM) did not affect hypoxic bronchodilation, suggesting that the release of intracellular Ca(2+) does not take a role in hypoxic bronchodilation. Hypoxic dilation decreased ATP content in epithelium-intact rings but not epithelium-denuded rings, indicating a relationship between hypoxic dilation and change of adenine nucleotide in epithelium-intact rings. Our findings indicate that the epithelium dependent mechanisms of hypoxic relaxation of guinea pig tracheal rings preconstricted with histamine may not be related to the mobilization of extra and intra-cellular Ca(2+).

Seasonal changes in the hypothalamo-pituitary-testes axis of the Japanese wood mice (Apodemus speciosus) were studied. The testes, epididymis, pituitary and hypothalamus were compared between mice in the breeding season (July) and non-breeding season (October) using morphological techniques, and the plasma testosterone level was evaluated by enzyme immunoassay. Significant differences in these tissues were observed between the breeding season and the non-breeding season. Specifically, differences in the non-breeding season included 1) a decline in testicular and epididymal weights, arrest of spermatogenesis and decrease of serum testosterone concentration; 2) a decrease in the number of luteinizing hormone (LH)-, follicle stimulating hormone (FSH)-, prolactin (PRL)-, and growth hormone (GH)-immunoreactive cells, and decrease in the size of FSH, PRL, and GH-immunoreactive cells; and 3) an increase in the size of gonadotropin-releasing hormone (GnRH)-immunoreactive neurons. Our findings indicate that the male adult Japanese wood mouse exhibits unique seasonal changes in the hypothalamo-pituitary-testes axis which are not found in laboratory mice. Anat Rec 260:366–372, 2000. © 2000 Wiley-Liss, Inc.

Silver plumage color mutant (B/B) quail has an abnormal retina characterizing the transdifferentiation of retinal pigment epithelium (RPE) following the retinal separation in the early developmental stage. In the present study; (i) the expression of gicerin, an immunoglobulin-superfamily cell adhesion molecule, was examined in the retina of B/B quail. In the wild-type quail, gicerin protein was enriched in the apical membrane (facing the neural retina, NR) of RPE cells on embryonic day (E) 4 and then appeared also in NR cells from E5. However, in the B/B retina, no gicerin expression was found in the transdifferentiation area of RPE prior to the retinal separation. (ii) In addition to this, microinjection of anti-gicerin polyclonal antibody into the eyeball of wild-type quail on E3 caused the retinal separation and induced the transdifferentiation of RPE into new NR. These observations suggest that the decrease of gicerin expression might participate in the retinal separation and RPE-transdifferentiation in B/B quail.

We examine sexual dimorphism in growth hormone-releasing hormone (GHRH) in the arcuate nucleus (ARC), and somatostatin (SS) in the periventricular nucleus (PeN) of the hypothalamus, and investigate when it becomes evident. Using immunohistochemical staining and morphometry, we observed ARC GHRH-immunoreactive (ir) neurons, ARC SS-ir neurons and PeN SS-ir neurons in male and female mice at 5, 20, 30, 40 and 60 days old. The number of ARC GHRH-ir neurons was significantly higher in males than females, after 20 days old. ARC SS-ir neurons showed no significant differences between sexes. On the other hand, PeN SS-ir neurons were significantly more numerous in males at 30, 40 and 60 days than in females. During postnatal development, these GHRH- and SS-ir neurons changed in different patterns from ages 20 to 60 days. The number of ARC GHRH-ir neurons in both sexes decreased from 5 to 20 days, increased until day 40, and then decreased at day 60, while ARC SS-ir neurons in both sexes increased from day 5 to day 60. PeN SS-ir neurons in both sexes increased from days 5 to 20 to 116% in males and 189% in females. Furthermore, in male mice, the increase continued until 40 days of age, while in females, there was no significant difference from days 20 to 60. There were no apoptotic cells; a few proliferating cell nuclear antigen (PCNA) stained cells were found in the ARC and PeN. Our results suggest that the sex difference of ARC GHRH neurons and PeN SS neurons appears by stimulation with testosterone during the development life. The developmental fluctuation in the number of ARC GHRH-ir neurons may not be modulated by testosterone, but by ARC SS neurons.

Quantitative changes of lung tissue components (air spaces lined by PAS-positive and PAS-negative epithelium, blood vessels and interstitium) were investigated in developing rats from fetal day 18 through neonatal day 1. The volume of the left lung increased significantly from fetal day 18 through neonatal day 1. The percentage and volume of the air spaces increased strikingly between fetal days 20 and 21. However, the percentage of the air spaces lined by PAS-positive epithelium decreased significantly from fetal days 20 to 21, and that of the spaces lined by PAS-negative epithelium increased between the two days. The proliferating cell nuclear antigen (PCNA)-positive cells were rich in the interstitium and epithelium of the air spaces on fetal days 18 and 19. The percentage of the interstitium decreased significantly from fetal day 18 through neonatal day 1, showing remarkable decrease between fetal days 20 and 21. From fetal day 20 onward, the PCNA-positive cells decreased in number and located in the epithelium of the conducting air ways and interstitium. Based upon these findings, the present study suggests that the period from fetal days 20 to 21 is a critical time for the development of fetal lung: the period before fetal day 20 is that for proliferation and the period after fetal day 21, functional differentiation of the lung.

In the murine uterus granulated metrial gland (GMG) cells appear only during normal pregnancy. GMG cells belong to a member of natural killer (NK) cells and play an important role in fetus survival and placental growth. Our previous study revealed that mouse GMG/uterine NK (uNK) cells in the late pregnancy rapidly disappear from the uterus, due to the degenerative change classified as necrosis. But there are few reports regarding appearance and morphology of uNK cells during late pregnancy. We examined histologically and histochemically how and when uNK cells undergo cell death. The uNK cells in the metrial gland increased in number and reached maximum until day 12 of pregnancy. Sudden disappearance, however, occurred after day 15 and the granules reduced in both number and size. In situ DNA fragmentation detection revealed that DNA fragmented uNK cells increased in number during days 13 to 15 and reached 70.2% at day 15 of pregnancy. From days 13 to 17, uNK cells were positive against anti-perforin antibody. Ultrastructurally, uNK cells at day 15 showed poor organelles and unusual granules in structure. In uNK cells at day 17, condensation of nucleus chromatin, reduction in size and phagocytosis into other uNK cells were observed. These results suggested that uNK cells undergo at least two types of cell death, classified as necrosis and apoptosis, at the different stages of pregnancy, and that perforin is not a mediator for cell death.

A line of transgenic rats (heterozygotes) carrying a chimeric gene comprising a regulatory portion of murine whey acidic protein and a structural portion of human GH (hGH) genes developed severe obesity with age. To characterize physiological mechanisms that lead to fat accumulation, an array of parameters related to obesity were studied. Blood hGH levels were continuously low, endogenous rat GH secretion was suppressed, and the pulsatility in peripheral GH levels was absent. Plasma glucose, insulin, triglyceride, and FFA levels in the male transgenic rats significantly exceeded those in nontransgenic littermates at 12 and 17 weeks, but not at 7 weeks, of age. All symptoms except hyperlipidemia were restored to normal by treatment with an antidiabetic agent, thiazolidinedione (troglitazone), for 1 week from 17 weeks of age. As phenotypic expression of obesity was already evident before aberration of physiological parameters, it was assumed that animals had a condition in which obesity or hyperlipidemia caused hyperinsulinemia. Gene expression and enzymatic activity of lipoprotein lipase in the adipose tissue in the transgenic rats were not different from those in normal rats. In contrast, the gene expression level of glycerol-3-phosphodehydrogenase was markedly elevated, suggesting that glycerol synthesis was much enhanced in the adipocytes of the transgenic rats. In an ip glucose tolerance test, the transgenic rats were not hyperglycemic at 7 weeks of age; however, the animal became hyperglycemic at 15–17 weeks of age. Finally, treatment with recombinant hGH for 1 week to produce pulsatile secretion reduced the size of epididymal and kidney fat pads and restored normal weight gain. These observations suggest that continuously low peripheral GH levels with the lack of pulsatile secretion resulted in obesity and noninsulin-dependent diabetes mellitus.

Background The pituitary gland of the dog is different from all animals and is described as “typical” for mammals. How might this complex pituitary gland of the dog be formed in fetal life? The current study examined the fetal development of the complex and unique dog pituitary and the ontogeny of specific cell types in the pars distalis. Methods Adenohypophysis of the beagle, from 25 to 60 days of gestation and at 2 days of age, was studied by immunocytochemical and histological staining. Results At 25 days gestation, the primordium (Rathke's pouch) of the adenohypophysis began to form by an upward evagination from the epithelium of the primary oral cavity. At 38 days, the pituitary gland showed the same morphology as in adult dogs, being merely smaller. Five walls of Rathke's pouch (anterior wall [A], lateral walls [L], posterior wall [P], and upper wall [U]) were found at 25 days, and by 38 days they had specialized into specific regions of the adenohypophysis through complex and unique processes. The pars intermedia was derived from the U and the dorsal area of the A. The pars tuberalis was derived from the dorsal area of the A. The pars distalis was formed by more singular processes: the peripheral areas of the pars distalis were first formed by A and P; then the ventral lumen of the extensive Rathke's lumen surrounded by these areas was filled up by proliferation of cells, although the dorsal lumen remains as Rathke's lumen after 38 days. The blood capillaries may play an important role in the development of parencymal cells in the Rathke's pouch during canine fetal life. At 30 days gestation, the first adrenocorticotropic hormone cells were found in the anterior- and posterior-ventral regions (derived from middle and ventral areas of the A and the P) of the pars distalis anlage, and blood capillaries invaded the parenchymal cells from the mesencyme surrounding the anlage. At 38 days, portal vessels without capillary loops in the median eminence had appeared, and growth hormone and luteinizing hormone cells appeared in the ventral areas of A and P in the pars distalis. By 52 days, when capillary loops were seen in the portal vessels in the median eminence, these types of cells spread through the whole pars distalis. Conclusion These areas in the epithelium of Rathke's pouch at 25 days may differentiate into specific regions of the pars distalis during subsequent fetal life, through complex processes that are characteristic to the canine species. Anat. Rec. 251:143–151, 1998. © 1998 Wiley-Liss, Inc.

Significant numbers of cells known by the name granulated metrial gland (GMG) cells, or uterine natural killer (NK) cells, are present in the murine uterus only during pregnancy. The functions of GMG cells remain incompletely defined, largely due to technical difficulties in isolating and culturing these cells. New methods for purifying large numbers of GMG cells are required. This study examined two methods for isolating GMG cells from the murine pregnant uterus based upon magnetic immunobeads and immunoparticles. By either approach, the yield of GMG cells was 1 × 104 cells/metrial gland. Isolations using immunobeads gave cell suspensions with 82% in purity and 70% in viability. Isolations using immunoparticles gave cell suspensions with 62% in purity and 67% in viability. GMG cells isolated by the magnetic method retained an ultrastructure similar to in vivo GMG cells. These results establish that immunomagnetic approaches are useful for isolation of GMG cells from the pregnant murine uterus.

Hyaluronidase (HAase) activity was detected in both a human pus sample and the culture supernatant of the only bacterial isolate from the pus, Streptococcus intermedius, using a zymographic technique. The optimum pH range for HAase activity was similar for both samples. Although the bands showing the strongest HAase activity of these samples differed from each other with respect to molecular size, both samples were equally inhibited by an antiserum raised against HAase of S. intermedius. These results suggest that S. intermedius may produce HAase in vivo as well as in vitro, and that this enzyme and/or its fragments may play an important role in host tissue degradation.

Background The present study was designed to clarify development of filtration property of fetal renal glomerulus when maternal kidney is dysfunctional. Materials and Methods Maternal bilateral ureteral ligation was performed on days 17, 19, and 21 of pregnancy. One day after each operation, cationized ferritin (CF), native ferritin (NF), and horseradish peroxidase (HRP) were injected, respectively, in the fetuses. Distribution of the tracers in the fetal kidneys was investigated electron microscopically. Results On fetal day 20, clustered CF particles were present in the laminae rarae interna and externa of the glomerular basement membrane of the fetuses from ureter-ligated mothers, while the clusters were arrayed in three to four layers in that of the fetuses from sham-ligated ones. On fetal day 22, a relatively large amount of CF particles was present in the lamina rara externa in the fetuses from ureter-ligated mothers when compared to that in the age-matched control fetuses. On fetal days 20 and 22, the number of NF particles was decreased, and shortening of the time for filtration of HRP through the glomerular basement membrane was observed in the fetuses from the ligated mothers. Conclusions These results suggest that dysfunction of maternal kidneys causes accelerated formation of fetal glomerular basement membrane and stimulates glomerular function in filtration in fetal rat kidney. Anat. Rec. 249:181–186, 1997. © 1997 Wiley-Liss, Inc.

The effect of continuous human GH (hGH) secretion on female reproduction was studied in adult female transgenic rats expressing the hGH gene with a mouse whey acidic protein (mWAP) promoter. Two lines of transgenic female rats carrying the mWAP/hGH gene were established and used in the study. One was characterized by relatively high levels of serum hGH (high line), and the other had relatively low levels (low line). High-line female rats had recurring, pseudopregnancy-like estrous cycles accompanied by increased serum progesterone levels for 2 wk after ovulation, and they were fertile. In these rats, luteinization occurred spontaneously without cervical stimulation, probably due to high levels of serum hGH, which has prolactin (PRL)-like activity in the rat. Although low-line female rats had recurring, regular 4-day estrous cycles, they were sterile. In these rats, pseudopregnancy could not be induced by mating or by mechanical cervical stimulation. PRL surges following cervical stimulation were not detected, and PRL secretion was not induced by a dopamine antagonist, metoclopramide. The ovarian tissue in this line had a much higher number of corpora lutea and grew much heavier than in normal littermates, suggesting impairment of PRL-induced structural luteolysis. Suppression of PRL secretion in low-line rats was, at least in part, due to a marked decrease in the number of lactotrophs in the pituitary. The present study shows that the serum hGH level plays a crucial role in regulating luteal function in female transgenic rats expressing the hGH gene.

Post-transfusion graft-versus-host disease (PT-GVHD) is a fatal adverse effect of blood transfusion.1)-3) The mortality rate is said to be 99% or higher.4) In spite of the seriousness of the disease, no definitively effective drug for curing patients of it has been identified. Recently, however, we reported on the potential usefulness of a protease inhibitor, nafamostat mesilate (NM), for treatment of PT-GVHD.5), 6) This potential usefulness of NM was suggested by the results in vitro experiments in which possible PT-GVHD effector cytotoxic T cell (CTL) clones and tumor necrosis factor β (TNF β) producing T cell clones established by us were used.7), 10) NM inhibited the cytotoxicity of the CTL clones and TNF β production by the TNF β producing clones.6) Based on these results, NM was administered to two patients with PT-GVHD. NM administration resulted in marked recovery of the patient peripheral blood mononuclear cell (PBMC) detected by microsatellite DNA polymorphism analysis distinguishable between donor and patient PBMCs. This is the first report of the marked recovery of PT-GVHD patient PBMC from very low ones which were replaced almost all with donor PBMCs. Moreover, NM administration was associated with body temperature normalization, disappearance of a body rash, liver function improvement and an unusual long survival time after onset of PT- GVHD in both cases. NM is potentially useful drug for treatment of PT-GVHD.

The pars intermedia of the pituitary gland, and plasma ACTH and cortisol levels in the pony, which was first diagnosed in Japan as indicating equine Cushing's disease, were examined by immunohistochemistry and radioimmunoassay, respectively. The pars intermedia was greatly enlarged and most of its cells were immunoreactive for antisera to both adenocorticotropic hormone (ACTH) and beta-endorphin (beta-End). The plasma ACTH level was elevated when clinical symptoms appeared. The present results reveal that equine Cushing's disease in this pony was induced by the hypersecretion of ACTH and beta-End from the enlarged pars intermedia of the pituitary gland.

In the current study, Cu2+ was tested for its ability to relax vessels and to accumulate cyclic GMP (cGMP) in rat pulmonary artery employing rat extrapulmonary arterial rings. Cu(2+)-induced relaxation was endothelium and concentration (in the range from 10(-7) to 10(-4) M) dependent. The content of cGMP in the rings was increased 1.7-fold with 10(-4) M Cu2+. NG-Monomethyl-L-arginine abolished both the copper-induced relaxation and the increase in cGMP of rings. Cu2+ and zaprinast, which inhibits phosphodiesterase activity, caused a synergistic increase in cGMP level in the rings, suggesting that Cu2+ enhanced cGMP level through a mechanism different from that of zaprinast, probably as a consequence of elevated accumulation of nitric oxide (NO). The magnitude of vasorelaxation observed due to simultaneous addition of Cu2+ and acetylcholine was additive, not synergistic. Cu2+ did not augment relaxation induced by exogenously added NO donor. These results suggest that Cu2+ elevates NO level in the rings not by prolonging the half-life of NO, but by activation of endothelial nitric oxide synthase and subsequently potentiating the action of NO on vascular tone.

The kidneys of fetal rats from uninephrectomized mothers were investigated morphometrically on days 18, 20, and 22 of gestation. Maternal uninephrectomy was performed on day 5. On days 20 and 22, the proximal tubular cell height in fetuses from uninephrectomized mothers was significantly lower than that in fetuses from sham-operated mothers. The luminal diameter, length and total volume of the proximal tubule were significantly larger in fetuses from uninephrectomized mothers than in those from sham-operated mothers. The total glomerular volume of mature type in fetuses from uninephrectomized mothers was significantly larger than that in fetuses from sham-operated mothers. On day 18, maternal uninephrectomy caused no significant difference in any kind of parameters used for the proximal tubule, but induced an increase in the total glomerular volume of immature type. These results suggest that maternal uninephrectomy stimulates morphological development of the proximal tubule in the fetal kidney.

Contrast agents have been shown to produce vasodilatory responses in several vascular beds. To our knowledge, however, their effect on the bronchial vasculature has not been examined. Clinically, contrast-induced bronchial vasodilation could potentially exacerbate life-threatening pulmonary hemorrhage during bronchial angiography for hemoptysis. In the current study, we systematically measured the bronchial vasodilatory response to diatrizoate meglumine 66% and diatrizoate sodium 10% (MD-76) and examined whether vasodilation would be mediated by nitric oxide (NO).We measured bronchial blood flow in seven anesthetized, ventilated, open-chested sheep using an ultrasonic flow probe placed around the bronchial artery. Bronchial blood flow was recorded before and after injection of 2 ml MD-76 into the bronchial artery. The protocol was repeated after 20 min infusion of N omega-nitro-L-arginine (L-NA; 10(-2) mol/l), an NO-synthase inhibitor, into the bronchial artery.There was a 45 +/- 8 ml/min increase (p.01) in bronchial blood flow after injection of MD-76, which was reduced to 20 +/- 6 ml/min (p.01) after infusion of L-NA.Bronchial arterial injection of MD-76 results in a consistent increase in bronchial blood flow that is mediated partly by NO.

To investigate how mitochondrial function was affected in leukotoxin (Lx)-,9,10-epoxy-12-octadecenoate-induced lung injury, lung mitochondria were extracted from isolated perfused rat lung with or without Lx-induced edematous injury. In the lung treated with 30 mumol of Lx, the mitochondrial respiration rate in states 3 and 4 significantly decreased (without mitochondrial uncoupling) concomitantly with increased release of lactate dehydrogenase (LDH), a parameter for cellular damage, into the perfusate and decreased ATP content in the lung tissue compared with those of untreated lung. Moreover, 30 mumol of Lx resulted in significant inhibition of cytochrome-c oxidase activity (vs. vehicle control). In contrast, lower doses of Lx (10 mumol) caused lung edema and cellular damage without evidence for mitochondrial dysfunction. We also examined cellular and mitochondrial damage in hydrostatic lung edema. Such edema showed neither suppressed mitochondrial respiration nor elevated LDH activity in perfusate, although lung wet weight increased as much as it did after 30 mumol Lx treatment. Our results suggest that the ex vivo mitochondrial dysfunction is one of the secondary (vs. initial augmented permeability) but specific manifestations of toxicity of Lx, and together with the previous reports, the ex vivo damaging effect of Lx against mitochondria may be ascribed not to its direct action on mitochondria but to Lx-derived cellular mechanism(s).

Persistent obstructive sleep apnoea syndrome (OSAS) occurs in approximately 20% of normal-weight children after adenotonsillectomy (TA) and, in nearly 70% of them, it is caused by adenoid regrowth. Patients with severe or moderate OSAS showed a high incidence of persistent disease even after TA. Allergic disease, severity and large adenoid size are associated with adenoid regrowth and persistent disease.To investigate factors contributing to persistent OSAS and adenoid regrowth after TA in normal-weight children.This was a prospective, observational study at a single institute and involved 49 normal-weight children with severe or moderate OSAS (apnoea-hypopnoea index, AHI, ≥ 5) who underwent TA. Background information, nasal endoscopic data and pre- and postoperative polysomnographic data were collected. A third polysomnography (PSG) was performed 1.5 year postoperatively in children who subsequently developed symptoms of sleep disturbance.Thirteen children (27%, 13/49) were symptomatic 1.5 years after TA. Allergic rhinitis (38.5% vs 11.1%, p = 0.03) and allergic disease (69.2% vs 30.6%, p = 0.02) were seen more frequently in these children. A third PSG confirmed persistent disease (AHI ≥ 5) in nine children (18.4%, 9/49). Six children (12.2%, 6/49) were diagnosed as having adenoid regrowth and three (6.1%, 3/49) underwent revision adenoidectomy.

Background Chronic cough is one of the most challenging symptoms to diagnose and treat, not only because of the variety of underlying disorders but also its varying susceptibility to treatments. Etiological studies of chronic cough vary depending on the clinical settings and the particular interests of investigators. Objectives The purposes of this study were first to categorize the etiology of chronic cough by its response to systematic diagnostic treatments starting from the β2 agonist and second to sub-categorize β2 agonist responsive cough (BRC) by the airway hyperresponsiveness. Methods One hundred and eighty-four never-smokers received the maximal dose of procaterol to diagnose BRC. BRC was sub-categorized into two groups with or without airway hyperresponsiveness measured by the methacholine challenge test. Sinobronchial syndrome (SBS) was diagnosed by postnasal drip symptoms and by the response to clarythromycin and carbocysteine. Atopic cough (AC) was diagnosed by the evidence of atopy and the response to cetirizine hydrochloride. Gastroesophageal reflux disease (GERD) was diagnosed by the response to rabeprazole sodium. Since we did not investigate eosinophil counts in the tissue or in the induced sputum, no diagnosis of eosinophilic bronchitis was made. Results One hundred and nine patients had BRC. Twenty-three of them had bronchial asthma (BA), 53 had cough variant asthma (CVA) and 33 had non-hyperresponsive BRC (NHBRC). Thirty-one patients had GERD, 27 had AC and 14 had SBS. Twenty-five patients had more than one diagnosis in combination, while 6 had other miscellaneous diseases. Twelve patients were undiagnosed and 11 dropped out of the study. Conclusions The majority of chronic cough was BRC. NHBRC was a new chronic cough entity. GERD is a common cause of chronic cough in Japan, as in Western countries. AC and SBS are also causes of chronic cough in Japan. Trial registration University hospital medical information network (UMIN 000007483)

Cyclic variation of heart rate (CVHR) associated with sleep apnea/hypopnea episodes has been suggested as a marker of sleep disordered breathing (SDB). This study examined the utility of ECG-based CVHR detection for diagnosing SDB using simultaneous polysomnography as the reference standard. We used a previously developed automated CVHR detection algorithm (autocorrelated wave detection with adaptive threshold, ACAT) that provides the number of CVHR per hour (CVHR index). The ACAT was refined using a polysomnographic database of 194 subjects with various severities of SDB and then, applied to a single channel ECG obtained during standard overnight polysomnography in 862 consecutive subjects referred for SDB diagnosis. Using multiple thresholds of CVHR index ≥38 and

Background: The neonatal administration of monosodium L-glutamate (MSG) has been used in investigations of the possible role of the arcuate nucleus in neuroendocrine regulation during postnatal development. We used this method to examine whether the mouse arcuate contained cell bodies immunoreactive with antisera to growth hormone releasing hormone (GHRH) or luteinizing hormone releasing hormone (LHRH), and whether these hypothalamic peptides affect synthesis and secretion of growth hormone and gonadotropin and the testis. Methods: The hypothalamus, pituitary, and testes of adult male mice treated with MSG during the neonatal period were fixed in Bouin's fluid or 10% neutral formalin. The hypothalamus was used in immune staining, the pituitary was used in both morphometry and immune staining, and the testis was stained with hematoxylin and eosin. Results: Body weights in control and treated mice were not different. The treated mice had more subcutaneous adipose tissue and a shorter body than the control mice. The testes were heavier in the controls. Many perikarya immunoreactive with antisera to GHRH or LHRH were found in the arcuate nucleus in control mice, but few such perikarya were found in this nucleus in treated mice. The size of the anterior lobe and the number and size of GH cells, follicle stimulating hormone (FSH) cells, and prolactin (PRL) cells in treated mice were less than those of control mice. Conclusions: GHRH and LHRH neurons in the arcuate nucleus in male mice may cause body and testis weight to increase via GH and LH cells, respectively, in the adenohypophysis during postnatal development. There are some differences in the hypothalamo-pituitary-testis axis of mice and rats. © 1994 Wiley-Liss, Inc.

Hypothalamic growth hormone-releasing factor (GRF) in higher mammals, including human GRF, is a 44 amino acid residue peptide and is highly homologous in structure. By contrast, mouse GRF (mGRF) recently deduced by cDNA cloning consists of only 42 residues and shows relatively low homology to the GRFs of higher mammals and the same rodent species, rat. To characterize and localize the predicted mature mGRF peptide in the hypothalamus, we have generated its antiserum and developed a homologous radioimmunoassay. Immunoreactive mGRF in the acid hypothalamic extract was eluted as a single peak at a position identical to that of synthetic peptide on both gel filtration chromatography and reverse-phase high-performance liquid chromatography (HPLC). Secretion of immunoreactive mGRF from incubated hypothalami increased several fold in response to 50 mM K+, and this rise was abolished in the absence of medium Ca2+. Only a single peak of immunoreactive mGRF that coeluted with synthetic replicate was observed after the K+ -stimulated medium was extracted on Bond Elut C18 cartridges and applied on reverse-phase HPLC. Immunohistochemistry identified many mGRF-positive cell bodies in the arcuate nucleus and dense bundles of immunoreactive fibers in the median eminence. Treatment of mice with gold thioglucose (GTG), a chemical agent known to cause hypothalamic lesions, markedly depleted both content and in vitro secretion of immunoreactive mGRF. The decline in mGRF secretion was greater in GTG obese than in nonobese mice, whereas somatostatin secretion was not affected by GTG treatment. As compared to the previous findings obtained on rat GRF, mouse had approximately the same content of hypothalamic GRF, but retained only one third the capacity to release GRF in response to the same depolarizing stimulus. These findings demonstrate that mouse hypothalamus contains and secretes a mGRF-like molecule which has immunological and chromatographic characteristics identical to that predicted by molecular cloning. The depolarization-evoked, Ca2+-dependent release of immunoreactive mGRF and its neuronal localization in the arcuate nucleus-median eminence region indicate that it functions as a hypophysiotropic hormone of physiological importance. GTG produces hypothalamic GRF deficiency, probably sparing hypophysiotropic somatostatin neurons, and could be used to generate a mouse model for human GRF-deficient dwarfism.

Wild Sika deer (Cervus nippon yesoensis) are seasonal breeders and their reproductive behaviors change annually. Since reproductive activity is regulated by gonadotropins, gonadotro-pic cells (LH and FSH cells) in pituitaries in 19 male and 18 female wild Sika deer, which were collected from Ashoro town in Hokkaido, were investigated by immunohistochemical methods. Gonadotropic cells remarkably changed between August (the end of non-mating season) and October (mating season); their distribution was wider and their immunostaining reactivity was more intensive in October than in August in males and females. We found that gonadotropic cells existed also in the pars intermedia and that these cells showed seasonal changes similar to those in the pars distalis. The significances of such seasonal changes in gonadotropic cells in the wild Sika deer were discussed.

The blood supply of the hypophysis in the dog was examined by light microscopy and scanning electron microscopy. The anterior hypophyseal arteries converged toward the infundibular stalk, subdivided in the stalk, and entered the primary capillary net and capillary loops in the median eminence lying at the base of the third ventriculus. The portal vessels originated from the capillary net and loops, and were continuous with the capillary sinusoids in the pars distalis. Some of the anterior hypophyseal arteries supplied the pars distalis directly and were also continuous with capillary sinusoids. In its caudal pole, the pars nervosa received the posterior hypophyseal arteries, which formed a capillary plexus in the pars nervosa. The vessels of the pars nervosa broke up into a primary capillary net at the caudal areas in the median eminence. The vasculature in the dog hypophysis is compared with that in the rat.

Adenohypophysis was immunocytochemically examined in an infertile female transgenic (Tg) mouse which carried human growth hormone (hGH) gene and had a high circulating level of hGH. No GH positive cells were detected. This confirmed the extrahypophyseal (ectopic) production of hGH and was coincident with the disappearance of parenchymal cells showing affinity to azocarmine in Azan staining. The normal frequency of ACTH positive cells was in accordance with the previous suggestion based on the changes found in zona fasciculata cells of the adrenal cortex. Most interesting findings were the detection of many PRL positive cells and the ovarian histology with nearly normal characteristics except for the presence of thick capsule and interstitial gland-like structure composed of large and light cells. Ovarian histology was also clearly different among individual Tg mice, even though they stemmed from the same line or progenitors, and had a similar phenotype. The current immunocytochemical observation well documented the changes revealed with Azan staining in the adenohypophysis about GH but not about PRL or ACTH. Thus, the immunocytochemical analysis of the adenohypophysis will provide useful methodology in assessment of endocrinological circumstances of Tg mice.

We report the case of a severe bronchial asthma attack 15 minutes after the ingestion of food containing small amounts of alcohol. Although an ethanol inhalation test was negative, an acetaldehyde inhalation test was positive. Furthermore, it was discovered that the patient was homozygous for a mutation of the aldehyde dehydrogenase-2 (ALDH-2) gene. We subsequently diagnosed his attack as acute severe alcohol-induced asthma. Since bronchial asthma patients who are homozygous for mutant ALDH-2 genes are susceptible to acute severe alcohol-induced asthma attacks, strict clinical attention is thought a necessity.

Background— Despite the adverse cardiovascular consequences of obstructive sleep apnea, the majority of patients remain undiagnosed. To explore an efficient ECG-based screening tool for obstructive sleep apnea, we examined the usefulness of automated detection of cyclic variation of heart rate (CVHR) in a large-scale controlled clinical setting. Methods and Results— We developed an algorithm of autocorrelated wave detection with adaptive threshold (ACAT). The algorithm was optimized with 63 sleep studies in a training cohort, and its performance was confirmed with 70 sleep studies of the Physionet Apnea-ECG database. We then applied the algorithm to ECGs extracted from all-night polysomnograms in 862 consecutive subjects referred for diagnostic sleep study. The number of CVHR per hour (the CVHR index) closely correlated ( r =0.84) with the apnea-hypopnea index, although the absolute agreement with the apnea-hypopnea index was modest (the upper and lower limits of agreement, 21 per hour and −19 per hour) with periodic leg movement causing most of the disagreement ( P Conclusions— The automated detection of CVHR with the ACAT algorithm provides a powerful ECG-based screening tool for moderate-to-severe obstructive sleep apnea, even in older subjects and in those with cardiac autonomic dysfunction.

Female mice were divided into androgenized (AF) and control (CF) groups. The AF mice were injected subcutaneously with testosterone propionate (Tp) and the CF mice with sesame oil at 5 days of age. Mammotropes (PRL cells) and somatotropes (GH cells) in the adenohypophyses of these mice when they became adults were studied with immunohistochemistry and morphometry by light microscopy correlated with routine electron microscopy. In CF mice, almost all of the PRL-immunoreactive cells (about 43% of all parenchymal cells) were type I (classical) PRL cells, and almost all of the GH-immunoreactive cells (about 30% of all parenchymal cells) were type I (classical) GH cells. Type II PRL cells accounted for about 0.4% of parenchymal cells, and type II GH cells were about 2.5% of all parenchymal cells. In AF mice, the percentages of PRL and GH cells were not significantly different from those of CF mice. Mammosomatotropes (Ms cells) in both groups were less than 1% of all parenchymal cells. Numbers of all parenchymal, PRL and GH cells, however, were increased significantly in AF mice when compared to those in CF, because the adenohypophysis was increased in volume in AF mice. Type I PRL cells were larger in AF than in CF. The ultrastructure suggested that type I PRL cells may show increased PRL synthesis and secretion in AF mice. Furthermore, AF mice, in which the hypothalamus is masculinized by the neonatal treatment with Tp, retained feminine characteristics in the population and size of PRL cells and GH cells in the adenohypophysis.

1) 進行肺癌症例に対する化学療法施行時にOK-432・5KE皮内投与 (週2～3回) を併用し, 骨髄抑制に対する効果を検討した.2) OK-432投与群では末梢血の白血球および顆粒球のNadirが非投与群と比較して有意に高かった.3) 小細胞肺癌例では化学療法施行後の血小板のNadirも非投与群と比較して有意に高かった.4) 白血球数2000/mm3未満, あるいは血小板数50000/mm3未1黄を示す症例はOK-432投与群で明らかに少なく, 更にその期間は短かった.5) 白血球のNadirから正常白血球数 (4000/mm3以上) に復するまでの期間はOK-432投与群が有意に短かった.