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1.
Jpn J Nurs Sci ; 21(1): e12581, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38146064

ABSTRACT

AIM: To clarify the state of screening and support systems for socially high-risk pregnant women at obstetric facilities across Japan and identify the characteristics of facilities related to the implementation of screening. METHODS: This cross-sectional study used a self-administered questionnaire. Participants were managers of hospitals, clinics, and midwifery birth centers handling deliveries in 47 prefectures across Japan. The questionnaire comprised items regarding the characteristics of participants and their facilities, service provision related to socially high-risk women available at the facility, the number of specified pregnant women (tokutei ninpu) per year, methods of screening, and support systems within the obstetric facilities. Descriptive statistics and multivariate logistic regression analysis were performed using IBM-SPSS version 24 for the association between facility characteristics and screening practices for socially high-risk pregnant women. RESULTS: Valid responses were received from 716 of 2512 obstetric facilities. Rates of specified expectant mothers per annual number of deliveries were identified as follows: perinatal medical centers (2.7%), general hospitals (1.6%), obstetrics and gynecology hospitals (1.0%), and clinics (0.8%). A total of 426 facilities (60.6%) reported screening all expectant mothers to identify socially high-risk pregnant women. Multiple logistic regression analysis revealed that facility characteristics and service/care provision related to screening practices included availability of in-hospital midwife-led care and in-hospital midwifery clinics (adjusted odds ratio 1.61; 95% CI [1.30, 1.47]), one-on-one care by midwife (1.73; 95% CI [1.15, 2.59]), multidisciplinary meetings within the facility (1.70; 95% CI [1.14, 2.56]), follow-up support systems after discharge (1.90; 95% CI [1.17, 3.09]), and participation in the regional council for children in need of protection (2.33; 95% CI [1.13, 4.81]). CONCLUSIONS: Approximately 60% of surveyed obstetric facilities screen for socially high-risk women. Increasing service provision at facilities may be necessary to implement screening.


Subject(s)
Obstetrics , Pregnancy, High-Risk , Child , Pregnancy , Female , Humans , Japan , Cross-Sectional Studies , Obstetrics/methods , Surveys and Questionnaires
2.
Anim Biosci ; 34(10): 1717-1722, 2021 Oct.
Article in English | MEDLINE | ID: mdl-33561326

ABSTRACT

OBJECTIVE: Stroking calves during the postnatal period could effectively improve humancalf relationships. The objective of this study is to examine how daily calf stroking frequency during the postnatal period affects the establishment of human-calf relationships. METHODS: Six calves were stroked by a trainer for 6 minutes once daily for 5 days after birth (D1). Six calves were stroked by a trainer for 3 minutes twice daily for 5 days after birth (D2). A further four calves were stared at but not stroked as the control group. The overall stroking or staring duration was the same for all groups, at 6 min/d and 30 min over 5 days. The tests for reactions to the stationary trainer in an unfamiliar environment and avoidance distance measurements for an approaching trainer were conducted at 1 month and 3 months after the treatment. RESULTS: Calves in both stroking groups approached significantly closer to the stationary trainer, vocalized less, and looked at the trainer shorter than the control group at 1 month. However, at 3 months, there was no significant difference between the D1 and the control group, whereas the D2 approached significantly closer to the trainer and vocalized less, and looked at the trainer for a shorter time than the control group. For the avoidance distance, the trainer could approach closer to both stroking groups than the control at 1 month, however, there was no difference among groups at 3 months. CONCLUSION: Our results suggested that the difference in the calf stroking procedure affected the established human-calf relationships, even though the total stroking duration was the same for all stroked calves. It is likely to be more effective to stroke more frequently than intensively when the aim is to establish better human-calf relationships within limited labor time.

3.
Biosci Biotechnol Biochem ; 78(3): 472-7, 2014.
Article in English | MEDLINE | ID: mdl-25036837

ABSTRACT

Mochi is highly cohesive and adhesive, and easy to choke on. Many of the fatal suffocation accidents with mochi occur in the elderly aged 65 years or older. These circumstances prompted us to investigate a special property of waxy wheat which is similar in texture to waxy rice, but is less cohesive and adhesive. We compared the differences in chewing and swallowing movements associated with eating waxy rice mochi and waxy wheat mochi between healthy adults and healthy elderly. Healthy elderly chewed mochi more and longer than healthy adults. Although there was no difference in the number of chewing cycles or total duration of chewing between the two types of mochi, waxy wheat mochi was easier to chew and left less pharyngeal residue. These findings lead us to suggest that waxy wheat mochi is promising as a food that is easy to swallow and difficult to choke on.


Subject(s)
Airway Obstruction/pathology , Eating/physiology , Food/adverse effects , Waxes/adverse effects , Aged , Aged, 80 and over , Endoscopes, Gastrointestinal , Humans , Japan , Oryza/adverse effects , Oryza/chemistry , Triticum/adverse effects , Triticum/chemistry , Waxes/chemistry
4.
Arch Phys Med Rehabil ; 95(5): 867-74, 2014 May.
Article in English | MEDLINE | ID: mdl-24055575

ABSTRACT

OBJECTIVE: To assess the jaw-opening force test (JOFT) for dysphagia screening. DESIGN: Criterion standard. SETTING: University dental hospital. PARTICIPANTS: Patients complaining of dysphagia (N=95) and with symptoms of dysphagia with chronic underlying causes (mean age ± SD, 79.3±9.61y; range, 50-94y; men: n=49; mean age ± SD, 77.03±9.81y; range, 50-94y; women: n=46; mean age ± SD, 75.42±9.73y; range, 51-93y) admitted for treatment between May 2011 and December 2012 were included. INTERVENTIONS: None. MAIN OUTCOME MEASURES: All patients were administered the JOFT and underwent fiberoptic endoscopic evaluation of swallowing (FEES). The mean jaw-opening strength was compared with aspiration (ASP) and pharyngeal residue observations of the FEES, which was used as the criterion standard. RESULTS: A receiver operating characteristic (ROC) curve analysis was performed. Forces of ≤3.2kg for men and ≤4kg for women were appropriate cutoff values for predicting ASP with a sensitivity and specificity of .57 and .79 for men and .93 and .52 for women, respectively. Based on the ROC analyses for predicting pharyngeal residue, forces of ≤5.3kg in men and ≤3.9kg in women were appropriate cutoff values, with a sensitivity and specificity of .80 and .88 for men and .83 and .81 for women, respectively. CONCLUSIONS: The JOFT could be a useful screening tool for predicting pharyngeal residue and could provide useful information to aid in the referral of patients for further diagnostic imaging testing. However, given its low sensitivity to ASP the JOFT should be paired with other screening tests that predict ASP.


Subject(s)
Deglutition Disorders/diagnosis , Deglutition/physiology , Exercise Test/methods , Exercise Therapy/methods , Jaw/physiopathology , Mass Screening/methods , Aged , Aged, 80 and over , Deglutition Disorders/physiopathology , Deglutition Disorders/rehabilitation , Female , Follow-Up Studies , Humans , Male , Middle Aged , Prognosis , ROC Curve , Reproducibility of Results
5.
Tohoku J Exp Med ; 231(3): 223-8, 2013 11.
Article in English | MEDLINE | ID: mdl-24240663

ABSTRACT

Swallowing disorders are common in the elderly, and aging is a factor that affects swallowing function. The elevation of the hyoid bone is important for swallowing and is controlled by the suprahyoid muscles. The hyoid and laryngeal elevation allows the bolus to enter the esophagus. The suprahyoid muscles, therefore, play an important role in swallowing. The effects of aging on suprahyoid muscle strength are unclear. In this study, we analyzed the effects of aging on suprahyoid muscle strength by comparing the jaw opening functions of healthy adults and elderly adults. The subjects were 150 healthy volunteers consisting of 76 adults aged < 70 years (38 men and 38 women; mean age, 48.8 ± 13.8 years; range, 23-69) and 74 elderly adults aged > 70 years (37 men and 37 women; mean age, 78.1 ± 4.8 years; range, 70-92). The jaw opening force (JOF) was measured with a jaw opening sthenometer and compared between the healthy adult and elderly groups. The mean JOF of healthy adults was about 10 kg in men and about 6 kg in women, which was significantly greater than the mean JOF of the healthy elderly subjects (about 7 kg in men and about 4 kg in women). The JOF of the men was significantly greater than that of the women in the healthy adult and elderly groups. We thus propose that aging decreases the strength of suprahyoid muscles in healthy adults. The swallowing function may decrease even in healthy adults, aged over 70 years.


Subject(s)
Aging/psychology , Deglutition/physiology , Hyoid Bone/physiology , Movement/physiology , Muscle Strength/physiology , Adult , Aged , Aged, 80 and over , Biomechanical Phenomena , Female , Humans , Jaw/physiology , Male , Middle Aged , Young Adult
6.
Hum Pathol ; 44(10): 2256-65, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23850497

ABSTRACT

Heme oxygenase-1 (HO-1) is a cytoprotective enzyme that is induced by intraplaque hemorrhage and degrades free heme and releases ferrous iron, which is rapidly sequestered by ferritin. In vitro studies have shown that binding of hemoglobin to hemoglobin scavenger receptor (CD163) induces HO-1 and the anti-inflammatory mediator interleukin (IL)-10. We immunohistochemically examined the relationship between CD163 expression in macrophages and intraplaque hemorrhage, HO-1, IL-10, and ferritin using coronary atherectomy specimens from patients with stable (SAP) or unstable angina pectoris (UAP). A total of 67 patients underwent atherectomy for SAP (n = 33) or UAP (n = 34). Samples were stained with antibodies against smooth muscle cells, macrophages, glycophorin-A (a protein specific to erythrocyte membranes), CD163, HO-1, IL-10, and ferritin. To identify cell types of HO-1-positive cells, double immunostaining was also performed. Double immunostaining for HO-1 and macrophages revealed that the vast majority of HO-1-positive cells were macrophages. Morphometric analysis demonstrated that CD163-positive macrophage score and the percentage of glycophorin-A-, HO-1-, IL-10-, and ferritin-positive areas were significantly higher in UAP than in SAP patients (CD163, P < .005; glycophorin-A, P < .0001; HO-1, P < .0001; IL-10, P < .005; ferritin, P = .0001). Moreover, CD163-positive macrophage score was positively associated with the percentage of glycophorin-A-, HO-1-, IL-10-, and ferritin-positive areas (glycophorin-A, r = 0.60, P < .0001; HO-1, r = 0.67, P < .0001; IL-10, r = 0.45, P < .0005; ferritin, r = 0.61, P < .0001). These findings suggest that enhanced expression of HO-1 and HO-1-related atheroprotective molecules plays an important role in exerting anti-inflammatory, antioxidant, and scavenging functions, which could contribute to plaque stabilization.


Subject(s)
Antigens, CD/metabolism , Antigens, Differentiation, Myelomonocytic/metabolism , Coronary Vessels/metabolism , Heme Oxygenase-1/metabolism , Plaque, Atherosclerotic/metabolism , Receptors, Cell Surface/metabolism , Receptors, Scavenger/metabolism , Atherectomy , Biomarkers/metabolism , Coronary Vessels/pathology , Female , Hemorrhage/complications , Hemorrhage/metabolism , Hemorrhage/pathology , Humans , Interleukin-10/metabolism , Male , Middle Aged , Plaque, Atherosclerotic/complications , Plaque, Atherosclerotic/pathology
7.
J Heart Valve Dis ; 22(5): 640-7, 2013 Sep.
Article in English | MEDLINE | ID: mdl-24383374

ABSTRACT

BACKGROUND AND AIM OF THE STUDY: Currently, there is an increased incidence of aortic valve stenosis (AS) in patients undergoing hemodialysis (HD), though the exact mechanisms are not fully understood. Myeloperoxidase (MPO) is a leukocyte-derived enzyme that catalyzes the formation of reactive oxygen species and is an index of oxidative stress. The study aim was to examine, immunohistochemically, the expression of MPO, using surgically resected aortic valve specimens from AS patients undergoing HD. METHODS: The study population consisted of 15 HD patients and 19 non-HD patients with severe AS undergoing aortic valve replacement. Frozen aortic valve samples obtained surgically from AS patients were stained immunohistochemically with antibodies against smooth muscle cells, neutrophils, macrophages, T lymphocytes, CD31, MPO and 4-hydroxy-2-nonenal (4-HNE). RESULTS: Quantitative analyses showed that the macrophage-positive area, and numbers of T lymphocytes, neutrophils, CD31-positive microvessels and MPO-positive cells in HD patients were significantly higher than in non-HD patients (macrophages, p < 0.0001; T lymphocytes, p < 0.0001; neutrophils, p < 0.0001; CD31, p < 0.0001; MPO, p < 0.0001). Moreover, the number of MPO-positive cells was positively correlated with CD31-positive microvessels and the 4-HNE-positive macrophage score (CD31, R = 0.73, p < 0.0001; 4-HNE, R = 0.49; p < 0.005). CONCLUSION: These findings suggest that MPO is highly expressed in the aortic valves of AS patients undergoing HD. Furthermore, MPO is positively associated with neovascularization and oxidative stress, which contribute to a rapid progression of AS in HD patients.


Subject(s)
Aortic Valve Stenosis/enzymology , Aortic Valve/enzymology , Oxidative Stress , Peroxidase/biosynthesis , Renal Dialysis , Aged , Aortic Valve/pathology , Aortic Valve Stenosis/complications , Aortic Valve Stenosis/pathology , Disease Progression , Female , Humans , Immunohistochemistry , Kidney Failure, Chronic/complications , Kidney Failure, Chronic/therapy , Macrophages/enzymology , Male , Neutrophils/enzymology , Retrospective Studies
8.
Osaka City Med J ; 59(2): 61-7, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24575581

ABSTRACT

BACKGROUND: Oxidative stress contributes to plaque formation and the destabilization of coronary atherosclerotic lesions. It has been reported that disease processes and clinical risk factors of aortic valve stenosis (AS) are similar to those of atherosclerosis. In this study, we immunohistochemically examined the expression of 4-hydroxy-2-nonenal (4-HNE), an oxidative stress-related molecule, by using surgically resected aortic valve specimens from AS patients. METHODS: The study was conducted using aortic valve specimens, surgically obtained from 24 patients with severe AS undergoing aortic valve replacement. We immunohistochemically investigated frozen aortic valve samples with antibodies against smooth muscle cells, macrophages, CD31 and 4-HNE. RESULTS: Morphometric analysis showed that the percentage of the macrophage-positive area and the number of CD31-positive microvessels were significantly higher in AS patients than those in reference cases (macrophages, p < 0.005 and CD31, p < 0.0001). Furthermore, the 4-HNE-positive macrophage score was also significantly higher in AS patients than in reference cases (p < 0.005). CONCLUSIONS: 4-HNE was expressed in the stenotic aortic valves in patients with severe AS, suggesting a close relationship between oxidative stress and the progression of calcific AS.


Subject(s)
Aortic Valve Stenosis/metabolism , Oxidative Stress , Aged , Aged, 80 and over , Aldehydes/analysis , Aortic Valve Stenosis/pathology , Female , Humans , Immunohistochemistry , Male , Platelet Endothelial Cell Adhesion Molecule-1/analysis
9.
Arch Phys Med Rehabil ; 93(11): 1982-6, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22676905

ABSTRACT

OBJECTIVE: To simplify the cough test to screen silent aspiration without sacrificing accuracy. DESIGN: Criterion standard. SETTING: University dental hospital. PARTICIPANTS: Consecutive patients (N=141; 92 men, 49 women; mean age 71±14y, range 23-94y) who had complained of some dysphagic symptoms between June 2008 and February 2010. INTERVENTIONS: All patients were administrated a simplified cough test and underwent the fiberoptic endoscopic evaluation of swallowing. Citric acid inhalation was terminated when the first cough occurred, and the time between the start of inhalation and the first cough was measured. MAIN OUTCOME MEASURES: The time when the first cough was observed by the simplified cough test was compared with the results of the fiberoptic endoscopic evaluation of swallowing, which was used as a criterion standard. RESULTS: Receiver operating characteristic curve analysis was performed for 53 patients evaluated as having aspiration by fiberoptic endoscopic evaluation of swallowing. We found that 30 seconds or less was an appropriate cutoff value for detecting patients without silent aspiration, where the sensitivity was .92 and the specificity was .94. From the receiver operating characteristic curve analyses for all patients, 60 seconds or less was determined to be an appropriate cutoff, and the sensitivity and specificity were .81 and .65, respectively. CONCLUSION: The simplified cough test is a useful screening tool for silent aspiration in patients with aspiration.


Subject(s)
Cough/chemically induced , Respiratory Aspiration/diagnosis , Adult , Aged , Aged, 80 and over , Citric Acid , Female , Hospitals, University , Humans , Male , Middle Aged , ROC Curve
10.
Arch Phys Med Rehabil ; 93(11): 1995-9, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22579648

ABSTRACT

OBJECTIVE: To investigate the effects of the jaw-opening exercise on decreased upper esophageal sphincter (UES) opening while swallowing. DESIGN: Intervention study: before-after trial with 4-week follow-up evaluation. SETTING: A university school of dentistry dental hospital. PARTICIPANTS: Patients with dysphagia (N=8; 7 men, 1 woman; average age ± SD, 70.5±11.3y; age range, 54-86y). INTERVENTIONS: All patients performed a jaw-opening exercise to strengthen the suprahyoid muscles. The exercise involved opening the jaw to its maximum and maintaining this position for 10 seconds. Each exercise set consisted of 5 repetitions, and 2 sets were carried out daily for 4 weeks. The effectiveness of the exercise was evaluated by a videofluorographic swallowing study (VFSS). MAIN OUTCOME MEASURES: Hyoid elevation, UES opening, pharynx passage time, and pharyngeal residue after swallowing at preexercise and postexercise were compared by VFSS. RESULTS: Compared with before starting the exercise, significant improvements were observed in the extent of upward movement of the hyoid bone (P<.05), the amount of UES opening (P<.05), and the time for pharynx passage (P<.05) 4 weeks after initiating the exercise. Pharyngeal residue decreased in some subjects, and no increases were noted in any subjects. CONCLUSIONS: The jaw-opening exercise is an effective treatment for dysphagia caused by dysfunction of hyoid elevation and UES opening.


Subject(s)
Deglutition Disorders/rehabilitation , Esophageal Sphincter, Upper/physiopathology , Exercise Therapy/methods , Jaw , Aged , Aged, 80 and over , Deglutition/physiology , Female , Hospitals, University , Humans , Male , Middle Aged
11.
J Nematol ; 43(1): 1-6, 2011 Mar.
Article in English | MEDLINE | ID: mdl-22791909

ABSTRACT

The effects of the non-fumigant nematicide imicyafos on soil nematode community structure and damage to radish caused by Pratylenchus penetrans were evaluated in two field experiments in consecutive years (2007 and 2008). Nematode densities in soil at 0 - 10 cm (the depth of nematicide incorporation) and 10 - 30 cm were measured. The application of imicyafos had a significant impact on the density of P. penetrans at 0 - 10 cm but had no effect on free-living nematode density. PCR-DGGE analysis conducted using extracted nematodes showed that the nematode community structure 12 d after application in 2007 was altered by the application of imicyafos at the 0 - 10 cm depth, but not at 10 - 30 cm. No significant differences were observed in the diversity of the nematode community at harvest (89 and 91 d after application) between the control and imicyafos treatments in both depths and both years. In both years, the damage to radish caused by P. penetrans was markedly suppressed by the nematicide. Overall, the nematicide imicyafos decreased populations of P. penetrans in soil and thereby decreased damage to radish, while having little impact on the soil nematode community.

12.
Somatosens Mot Res ; 27(3): 93-9, 2010.
Article in English | MEDLINE | ID: mdl-20653500

ABSTRACT

We studied changes in orofacial behavior and the arrangement of bilateral hypoglossal nuclei after the neurectomy of the medial branch of the unilateral hypoglossal nerve in cats. After recovery from surgery in a head holder, the animals were acclimated to take and chew fish paste (1.8 g) from a spoon and lick milk from a wetted paintbrush. Next we performed a neurectomy in the unilateral hypoglossal nerve after training. We firstly recorded behavior during the taking of fish paste and licking of milk, and then performed a neurectomy in the unilateral hypoglossal nerve. After nerve cutting, the cats' tongue deviated toward the cut side when they licked food, and bilateral activities of EMGs in the genioglossus muscles became stable in about 1 month. After that, we injected two kinds of fluorescent dye (10% Evans blue, EB, and 3% Fast blue, FB) into the bilateral genioglossus muscles using syringes (0.15 ml in each), respectively. Although each injection of FB and EB into the bilateral genioglossus muscles in normal cats revealed cells positively stained with each dye in the hypoglossal nuclei of each injection site, in cats 1 month after nerve cutting, fluorescent dye was only observed in positive cells in the hypoglossal nucleus of the intact side and the dye injected into the neurectomy side showed a mixture into positive cells of the intact side. The findings suggest that muscles in the neurectomy side may be compensated by regeneration of the peripheral nerves on the intact side.


Subject(s)
Hypoglossal Nerve Injuries , Medulla Oblongata/physiology , Motor Neurons/physiology , Tongue/innervation , Animals , Cats , Female , Fluorescent Dyes , Male
13.
Gan To Kagaku Ryoho ; 37 Suppl 2: 159-61, 2010 Dec.
Article in Japanese | MEDLINE | ID: mdl-21368509

ABSTRACT

Dysphagia can cause aspiration pneumonia. The condition of dysphagia is difficult to evaluate from outside. Therefore, a careful examination is necessary to grasp the state of swallowing of a patient accurately. However, it has been a difficult situation for a patient who cannot come to hospital for some reason to be examined by video fluoroscopy or video endoscopy. In recent years, a usefulness of video endoscopy in visiting home examination for dysphagia has been reported several times. And this video endoscopy examination is a valuable tool to detect a discrepancy between swallowing function and nutritional intake of the patient. Cooperative rehabilitation with such a careful examination is an important issue to be successful in dysphagia rehabilitation.


Subject(s)
Deglutition Disorders/rehabilitation , Endoscopy/methods , Home Care Services , Aged , Deglutition , Deglutition Disorders/physiopathology , Endoscopy/instrumentation , Female , Humans
14.
Acta Crystallogr Sect E Struct Rep Online ; 64(Pt 9): m1110, 2008 Aug 06.
Article in English | MEDLINE | ID: mdl-21201575

ABSTRACT

In the title compound, [Ru(C(44)H(28)N(4))(C(8)H(5)F(3))], the fluorin-ated alkyl-idene group is bound to a five-coordinate Ru atom, which is located toward the carbene C atom, 0.3301 (5)Šfrom the least-squares plane of the C(20)N(4) porphyrin core. The Ru=C bond is tilted slightly from the normal to the C(20)N(4) least-squares plane due to steric repulsion between the porphyrinate ligand and the bulky trifluoro-methyl group. The Ru=C bond length of 1.838 (2) Šis comparable with those in bis-(subsituted phen-yl)carbene analogs.

15.
Microbes Environ ; 23(4): 331-6, 2008.
Article in English | MEDLINE | ID: mdl-21558727

ABSTRACT

The toxicity of three organophosphorous nematicides, imicyafos, fosthiazate and cadusafos, to non-target organisms in soil was evaluated. Imicyafos and fosthiazate had no significant inhibitory effect on the growth of fungal (Fusarium oxysporum f. sp. lactucae, Rhizoctonia solani and Trichoderma viride) and bacterial (Ralstonia solanacearum and Pseudomonas fluorescens) strains in media at 12.5 to 200 mg L(-1). Cadusafos, however, significantly inhibited the growth of all these strains except R. solanacearum. A pot test was conducted using a soil naturally infested with Pratylenchus penetrans, and treated with imicyafos or fosthiazate, which are less toxic to non-target organisms. The density of P. penetrans decreased to less than 10% of the control level after exposure to imicyafos and fosthiazate at 3 kg active ingredient ha(-1), the conventional dose. No significant effect was observed on the density of free-living nematodes, cellulose decomposition activity, microbial biomass evaluated with the ATP method and number of ammonia oxidizers between the soil treated with imicyafos or fosthiazate and the untreated control soil. Our results revealed that imicyafos and fosthiazate effectively suppressed a plant-parasitic nematode, P. penetrans, but had little impact on free-living nematodes and the soil microbial community.

16.
Biochemistry ; 42(47): 13977-86, 2003 Dec 02.
Article in English | MEDLINE | ID: mdl-14636066

ABSTRACT

Cholesterol has been claimed to be involved in the generation and/or accumulation of amyloid beta protein (Abeta). However, the underlying molecular mechanisms have not been fully elucidated yet. Here, we have investigated the effect of membrane cholesterol content on gamma-secretase activity using Chinese hamster ovary cells stably expressing beta-amyloid precursor protein (APP) and either wild-type or N141I mutant-type presenilin 2. Cholesterol was acutely depleted from the isolated membrane by methyl-beta-cyclodextrin, and Abeta production was assessed in a cell-free assay system. Reduced cholesterol did not significantly alter the amounts of Abeta produced by either total cell membranes or cholesterol-rich low-density membrane domains. Even its extremely low levels in the latter domains did not affect Abeta production. This indicates that the membrane cholesterol content does not directly modulate the activity of gamma-secretase. To ascertain that gamma-secretase resides in cholesterol-rich membrane domains, low-density membrane domains were further fractionated with BCtheta (biotinylated theta-toxin nicked with subtilisin Carlsberg protease), which has recently been shown to bind selectively to rafts of intact cells. The membrane domains purified with BCtheta did indeed produce Abeta. These observations indicate that the gamma-cleavage required for generating Abeta occurs in rafts, but its activity is virtually cholesterol-independent.


Subject(s)
Amyloid beta-Protein Precursor/metabolism , Cholesterol/physiology , Endopeptidases/metabolism , Membrane Microdomains/enzymology , beta-Cyclodextrins , Amyloid Precursor Protein Secretases , Amyloid beta-Peptides/antagonists & inhibitors , Amyloid beta-Peptides/biosynthesis , Amyloid beta-Protein Precursor/antagonists & inhibitors , Amyloid beta-Protein Precursor/biosynthesis , Amyloid beta-Protein Precursor/chemistry , Animals , Anticholesteremic Agents/chemistry , Aspartic Acid Endopeptidases , Bacterial Toxins/chemistry , Biotinylation , CHO Cells , Cholesterol/metabolism , Cricetinae , Cyclodextrins/chemistry , Endopeptidases/chemistry , Hemolysin Proteins , Humans , Membrane Microdomains/chemistry , Membrane Microdomains/metabolism , Peptide Fragments/chemistry , Peptide Fragments/metabolism , Protein Structure, Tertiary , Solubility
17.
Acta Crystallogr C ; 59(Pt 9): m369-70, 2003 Sep.
Article in English | MEDLINE | ID: mdl-12944647

ABSTRACT

The stable title trifluoromethyl-substituted carbenyl metalloporphyrin, [Ru(C(15)H(8)F(6))(C(48)H(36)N(4))], has a five-coordinate Ru atom which is displaced from the porphyrin N(4) plane towards the axial carbene ligand by 0.230 (3) A. The Ru--C(carbene) bond coincides with a crystallographic twofold axis and its length of 1.841 (6) A is notably shorter than the value of 1.868 (3) A in the pyridine adduct.

18.
Acta Crystallogr C ; 59(Pt 2): m37-9, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12574641

ABSTRACT

In the two ruthenium(II)-porphyrin-carbene complexes (dibenzoylcarbenyl-kappaC)(pyridine-kappaN)(5,10,15,20-tetra-p-tolylporphyrinato-kappa(4)N)ruthenium(II), [Ru(C(15)H(10)O(2))(C(5)H(5)N)(C(48)H(36)N(4))], (I), and (pyridine-kappaN)(5,10,15,20-tetra-p-tolylporphyrinato-kappa(4)N)[bis(3-trifluoromethylphenyl)carbenyl-kappaC]ruthenium(II), [Ru(C(15)H(8)F(6))(C(5)H(5)N)(C(48)H(36)N(4))], (II), the pyridine ligand coordinates to the octahedral Ru atom trans with respect to the carbene ligand. The C(carbene)-Ru-N(pyridine) bonds in (I) coincide with a crystallographic twofold axis. The Ru-C bond lengths of 1.877 (8) and 1.868 (3) A in (I) and (II), respectively, are slightly longer than those of other ruthenium(II)-porphyrin-carbene complexes, owing to the trans influence of the pyridine ligands.

19.
J Reprod Dev ; 49(4): 313-21, 2003 Aug.
Article in English | MEDLINE | ID: mdl-14967924

ABSTRACT

To reveal the molecular mechanism of selective follicular atresia in porcine ovaries, we investigated the changes in the expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptor (DR4) proteins and TRAIL mRNA in granulosa cells during follicular atresia. Immunohistochemical, Western immunoblotting and reverse transcription-polymerase chain reaction analyses (RT-PCR) revealed that significant increases in TRAIL protein and mRNA levels but not DR4 protein were changed during atresia. The RT-PCR product was confirmed to be porcine TRAIL by the cDNA sequence determination. An in vitro apoptosis inducing assay using cultured granulosa cells prepared from healthy follicles showed that TRAIL could activate caspase-3 and induce apoptotic cell death in the cells. The present findings confirm that TRAIL induces apoptosis in granulosa cells during atresia in porcine ovaries.


Subject(s)
Apoptosis/physiology , Follicular Atresia/physiology , Granulosa Cells/cytology , Membrane Glycoproteins/physiology , Ovary/cytology , Tumor Necrosis Factor-alpha/physiology , Animals , Apoptosis Regulatory Proteins , Base Sequence , Caspase 3 , Caspases/metabolism , DNA, Complementary/genetics , Enzyme Activation , Female , Follicular Atresia/genetics , Gene Expression , Granulosa Cells/physiology , In Vitro Techniques , Membrane Glycoproteins/genetics , Ovary/physiology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, TNF-Related Apoptosis-Inducing Ligand , Receptors, Tumor Necrosis Factor/physiology , Signal Transduction , Sus scrofa , TNF-Related Apoptosis-Inducing Ligand , Tumor Necrosis Factor-alpha/genetics
20.
J Vet Med Sci ; 64(5): 435-9, 2002 May.
Article in English | MEDLINE | ID: mdl-12069077

ABSTRACT

Previously, we histochemically examined the localization of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and its receptors in porcine ovarian follicles, and demonstrated a marked reduction in the expression of TRAIL-decoy receptor-1 (DcRI) in granulosa cells of atretic follicles. In the present study, to confirm the inhibitory activity of DcR1 in granulosa cells, granulosa cells prepared from healthy follicles were treated with phosphatidylinositol-specific phospholipase C (PI-PLC) to cleave glycophospholipid anchor of DcR1 and to remove DcR1 from the cell surface, and then incubated with TRAIL. PI-PLC treatment increased the number of apoptotic cells induced by TRAIL. The present finding indicated the possibility that TRAIL and its receptors were involved in induction of apoptosis in granulosa cells during atresia, and that DcR1 plays an inhibitory role in granulosa cell apoptosis.


Subject(s)
Apoptosis , Granulosa Cells/cytology , Granulosa Cells/metabolism , Ovarian Follicle/cytology , Receptors, Tumor Necrosis Factor/metabolism , Animals , Apoptosis/drug effects , Apoptosis Regulatory Proteins , Cells, Cultured , Female , GPI-Linked Proteins , Granulosa Cells/drug effects , Membrane Glycoproteins/antagonists & inhibitors , Membrane Glycoproteins/pharmacology , Phosphatidylinositol Diacylglycerol-Lyase , Phosphoinositide Phospholipase C , Receptors, Tumor Necrosis Factor, Member 10c , Swine , TNF-Related Apoptosis-Inducing Ligand , Tumor Necrosis Factor Decoy Receptors , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Tumor Necrosis Factor-alpha/pharmacology , Type C Phospholipases/metabolism
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