In the past decade, multidrug-resistant Pseudomonas aeruginosa (MDRP) infection has become a serious clinical problem, due to the limitation of drug choices to fight against the bacteria. Here we explored the bactericidal activity in the filtrated supernatant of Streptococcus (S.) sanguinis against Pseudomonas (P.) aeruginosa. S. sanguinis is one of the α-hemolytic streptococci that commonly reside in the human oral cavity. A strain of S. sanguinis, isolated from the sputum of a pulmonary-disease patient, was cultured for overnight. The filtered supernatant was tested for bactericidal effect using the minimum bactericidal concentration method on 20 strains of P. aeruginosa, including two MDRP and five mucoid-type strains. The viable number of P. aeruginosa was decreased with time after exposing to the filtrated supernatant of S. sanguinis, and collapsed bacteria were detected with electron microscopy. Of the 20 strains, 19 (95%) strains of P. aeruginosa were affected by bactericidal effect. Among other species of bacteria examined, the filtrated supernatant of S. sanguinis showed remarkable bactericidal effect on 49% of indole-positive Proteus species (4/9 strains) and 60% of Acinetobacter (A.) baumannii (6/10 strains). We next investigated the property of bactericidal activity in filtrated supernatant by treating with proteinase K or autoclave. There was no change in the bactericidal activity of the filtrated supernatant after each treatment, excluding the involvement of protein and plasmid. Here, we identify the bactericidal activity in the filtrated supernatant of S. sanguinis against MDRP. This unexpected observation may contribute to the development of a novel therapeutic drug against P. aeruginosa.
Longitudinal studies in Japan indicate that nocturnal sleep onset has become later and sleep duration has been progressively shortened. This study aimed to investigate the relationship between sleep patterns and sleep problems among children, and to determine the association between parents and their children's sleep habits. Questionnaires about sleep problems and life habits were administered to families living in Tokyo metropolitan areas of Japan. We analyzed the data of pre-school-age (1-5 years old; n = 319, including 175 girls) and elementary school-age children (6-11 years; n = 217, including 116 girls) as well as their parents (402 mothers: 37.0 ± 4.9 years, 402 fathers: 39.0 ± 5.9 years). Subjects were categorized as morning (evening) type when they answered their lifestyle habit as “definitely or moderately morning (evening) type”. Sleep was categorized into regular, irregular, and intermediate from the sleeping-waking regularity scores. The frequency of daytime dozing is significantly high in children with evening-irregular sleep. Moreover, mothers of children (aged 1-5 and 6-11 years) with evening-irregular sleep have significantly more irregular sleep habits than those of children with morning-regular sleep. Likewise, fathers of children (aged 1-5 years) with evening-irregular sleep have significantly more irregular sleep habits. Thus, irregular late bedtime of parents is associated with sleep problems, daytime sleepiness, and irregular dietary habits of children. Mothers' sleep habits have a stronger influence on their children's sleep than fathers'. Our study indicates the importance of promoting sleep hygiene that encourages healthy sleep for all family members.
Knee pain is extremely common among the elderly, particularly women. Hence, there is an urgent need for applicable community-based intervention models for halting the progression of knee pain and related disabilities in elderly women. We aimed to assess the efficacy of home-visit physiotherapy as a new intervention model. This non-randomized 5-month-long controlled trial enrolled elderly community-dwelling women (aged 60-83 years) with mild knee pain. The intervention consisted of two home visits by a physiotherapist, with instructions on routinely performing muscle-strengthening exercises at home and implementing simple environmental modifications when necessary. Outcome measures were assessed at baseline and 5 months later. The primary outcomes were measured as the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and a newly devised seiza-style sitting score. People in Japan, especially elderly women, are accustomed to seiza-style sitting that involves kneeling on one's lower legs while resting the buttocks on the heels. The secondary outcomes included quadriceps isometric strength, knee alignment in the frontal and sagittal planes, and passive knee extension range. Forty-two subjects (20 in the intervention group and 22 in the control group) completed the study. At baseline, characteristics and the primary outcomes did not significantly differ between the 2 groups. At 5 months, the primary outcomes improved significantly in the intervention group. Estimated differences in the change from baseline for each outcome between the 2 groups were computed, adjusting for outcome variables imbalanced at baseline. Even after the adjustment, the home-visit physiotherapy regimen provides favorable improvement in the seiza-style sitting score.
Large vessel vasculitis leads to arterial wall thickening and stiffening because of chronic inflammatory changes. The cardio-ankle vascular index (CAVI) is recently utilized for assessing arterial stiffening caused by atherosclerosis-related diseases, including hypertension and diabetes, as well as aging. CAVI is mathematically calculated from stiffness index beta, which is established as a parameter of arterial stiffness independent of blood pressure. However, there are no data regarding arterial stiffness assessed by CAVI for large vessel vasculitis. We describe a patient with large vessel vasculitis who showed aortic wall thickening and increased CAVI without hypertension. A 68-year-old woman presented at our hospital with recurrent fever of 2-month duration, fatigue, neck pain, and weight loss. The images of 18F-fluorodeoxyglucose-positron emission tomography (18F-FDG-PET) demonstrated significant 18FDG uptake (indicating increased metabolic activity and presence of inflammation) in the aorta and its major branches, including the carotid and subclavian arteries. Contrast-enhanced magnetic resonance imaging demonstrated wall thickening of the thoracic aorta. These imaging findings resulted in the diagnosis of large vessel vasculitis. The patient showed normal brachial blood pressure (right, 122/72 and left, 121/66 mmHg). However, CAVIs on both sides (right, 10.3 and left, 10.4) were elevated (normal value for her age, 9.1 ± 0.8). In conclusion, arterial stiffness in patients with large vessel vasculitis may be increased because of the arterial wall thickening and inflammatory changes. Thus, CAVI may be promising for detection of increased arterial stiffness in patients with large vessel vasculitis in the early stage, in which blood pressure is normal.
The therapy for acute myocardial infarction (AMI) has been improved; yet, AMI remains a major cause of death and heart failure in industrialized countries. B-type natriuretic peptide (BNP), a hormone secreted from the heart, has been shown cardioprotective effects during myocardial ischemia/reperfusion. In the present study, we aimed to examine whether BNP could inhibit myocardial apoptosis during ischemia/reperfusion. Rabbits were randomly divided into three groups (12 animals for each group): sham-operated control and ischemia-reperfusion animals with or without BNP treatment. Occlusion of the left circumflex coronary for 45 min was followed by 3-h reperfusion with infusion of physiological saline (untreated group) or BNP (treated group) starting 5 min before reperfusion and throughout the whole reperfusion. The infarct size, measured by triphenyltetrazolium chloride staining, was reduced by 44% with BNP treatment (P < 0.01). Accordingly, serum levels of creatine kinase and lactate dehydrogenase were markedly reduced in BNP-treated group (P < 0.05) compared with the untreated group. BNP significantly attenuated apoptotic cells (TUNEL-positive cardiomyocyte nuclei) in the myocardium (P < 0.01). The BNP-mediated attenuation of apoptosis was associated with the increased expression of an anti-apoptotic protein Bcl-2 and the reduced expression of a pro-apoptotic protein Bax. Moreover, BNP treatment significantly decreased the magnitude of caspase-3 activation caused by myocardial ischemia-reperfusion. In conclusion, pretreatment with BNP shortly before the onset of reperfusion not only reduces necrosis, but also attenuates myocardial apoptosis. BNP appears to be an ideal pharmacological agent applied as an adjuvant therapy to current myocardial reperfusion strategies.
Regeneration of segmental bone defects has been a clinical challenge. Recent advances in the field of tissue engineering have developed new procedures enabling bone regeneration. Small animal models capable of supporting weight-bearing femoral defects are integral parts of orthopedic biomedical research. However, a drawback of bone healing research is the lack of stable and adaptable fixation devices for small animals. Therefore, we developed and evaluated an adjustable external fixation device in the maintenance of non-healable (ie critical-sized) segmental defects, and in the fixation of tissue-engineered bone grafts in a rat model. Male Sprague-Dawley rats (n = 24) underwent a femoral osteotomy to create a non-healing segmental defect (6 mm size), which was stabilized with the fixator. A treatment group (12 rats) received tissue-engineered bone graft implants consisting of biphasic calcium phosphate blocks seeded with bone mesenchymal stem cells, while other 12 animals received no bone graft (non-treatment group). The osteotomy gap remained unchanged in the non-treatment group over the 12-week period, indicating that the 6-mm bone defect is really non-healable in the rat femur and that the device has sufficient stability for the management of critical-sized femoral defects. At 12 weeks, the treatment group maintained the bone length throughout the study period and showed bridging of the defect, with remarkable new bone formation. In contrast, the non-treatment group showed marginal new bone formation, but no apparent healing. In conclusion, the novel device provides substantial benefits in the maintenance of critical-sized femoral defects and tissue-engineered bone grafts in a rat model.
The most frequent cause of death in hemodialysis patients is cardiovascular disease with chronic inflammation being an epidemiologically proved risk factor. Many studies have shown C-reactive protein (CRP) as the strongest predictor of long-term mortality of hemodialysis patients, while other reports have indicated acute phase proteins as potential predictors of the mortality. The present study therefore aimed to evaluate the prevalence of chronic inflammation in hemodialysis patients and the role of acute phase proteins together with lipids and divalent ions for predicting mortality in hemodialysis patients. Chronic inflammation was defined, based on the serum level of high sensitive CRP > 8.4 mg/L and/or serum amyloid-A (SAA) > 8.9 mg/L. Acute phase proteins are defined as one whose plasma concentration increase (positive) or decreases (negative) by at least 25% during inflammation. High sensitive CRP and SAA were positive acute phase proteins measured, while albumin and fetuin-A, a calcification inhibitor, were selected as negative acute phase proteins. This prospective 36-month follow-up study included 130 patients (60 males and 70 females, aged 55.1 ± 12.9 years) maintained by hemodialysis for 107.2 ± 54.72 months at a Nephrology Clinic in Belgrade. The prevalence of chronic inflammation was 35.4% (46 patients). During the follow-up period, 24 patients (18.5%) died and 2 patients received transplants. In multivariate analysis, potential independent predictors of mortality in hemodialysis patients are hyperphosphatemia, hypoalbuminemia, and high SAA. Considering that assays for SAA are widely used, we propose that SAA is the best predictor for outcomes of end-stage renal disease.
For the patients with severe hemiplegia, long-time wheelchair sitting is unavoidable, which however increases a risk of secondary impairments due to non-use of the affected leg. A cycling wheelchair (C-W/C) has a possibility to activate paretic muscle through self-locomotion with bilateral pedaling. We therefore measured driving speed of C-W/C and electromyogram (EMG) in both legs during driving in the healthy adults and severe hemiplegic patients. Ten healthy volunteers (mean age 32.8, 26-45 years) and ten non-ambulatory post-stroke patients (mean age 69.0, 55-81 years) with complete or semi-complete hemiplegia participated in this study. EMG was recorded from the key muscles for cycling during isometric movement as baseline and during driving a C-W/C straightforward. All of the patients could drive a C-W/C with mean maximum driving speed of 46.6 (31.7-61.7) m/min, which was about half of that in the healthy subjects and within practical level. Root mean square of EMG (R-EMG) as a parameter reflecting muscle activity was compared between baseline and C-W/C driving. There was no increase in most of the values of R-EMG during driving in the healthy subjects and in the intact side of the hemiplegic patients. In contrast, significant increase was found during driving in several paretic muscles, despite that EMG of the paretic leg showed almost silent at baseline. These results suggest C-W/C can induce muscle activities of the paretic leg and provide a chance of practical locomotion even for the severe hemiplegics. Daily use of a C-W/C may contribute to restore paretic leg function.
Running exercise is an effective therapy for the prevention of osteoporosis; however, appropriate duration of exercise has not been determined. We therefore investigated the effect of exercise duration on bone mineral density (BMD) and systemic bone metabolism using young growing rats. Fifteen 8-week-old female Wistar rats were divided into three groups according to running load: control group (no running), short duration (30 min/day) and long duration (180 min/day), and animals ran on a treadmill 5 days per week over an 8-week period. BMD of the tibia was measured using peripheral quantitative computed tomography, and serum levels of tartarate-resistant acid phosphatase (TRAP), a bone resorption marker and alkaline phosphatase (ALP), a bone formation marker were measured to know whether the treadmill exercise would affect systemic bone metabolism. Short-duration running exercise (30 min/day) caused a significant increase in BMD of the metaphyseal trabecula (p < 0.05) with a reduction of serum TRAP levels (p < 0.01) and an increase in serum levels of calcium (p < 0.05) and phosphorus (p < 0.01). Conversely, long-duration exercise (180 min/day) significantly reduced BMD of the diaphyseal and metaphyseal cortex and that of the diaphyseal trabecula with a significant reduction of serum ALP levels and a significant increase in serum phosphorus. These findings suggest that short-duration exercise may increase BMD through suppression of bone resorption, whereas long-duration exercise may reduce BMD through suppression of bone formation. Exercising for short duration but not prolonged exercise is recommended to increase BMD of loaded long bones.
Lipoatrophy is the long-term adverse effects developed in human immunodeficiency virus (HIV)-1-infected subjects receiving highly active antiretroviral therapy (HAART). This cross-sectional study aimed to evaluate the prevalence of and clinical factors associated with lipoatrophy in HIV-infected Koreans receiving HAART for more than 6 months. Lipoatrophy was diagnosed by concordance between physical examination and history taking performed by a single physician. Various covariates were examined, including diabetes mellitus (DM), lipid profiles after HAART, and HAART regimen and duration. Among total 144 patients (6 females and 138 males), 35 patients (24.3%) were diagnosed with lipoatrophy. The prevalence of lipoatrophy was significantly higher in females than that in males [83.3% (5/6) vs. 21.7% (30/138), p = 0.010] and higher in patients with DM than patients without DM [66.7% (4/6 DM) vs. 22.5% (31/138 non-DM), p = 0.030], or in patients with high total cholesterol levels than patients with low total cholesterol levels [31.9% (23/72 patients with high cholesterol) vs. 16.7% (12/72 patients with low cholesterol), p = 0.035]. Moreover, patients with stavudine treatment history (> 12 months) had a higher prevalence of lipoatrophy than patients who never received stavudine [50.0% (15/30) vs. 16.5% (17/103), p < 0.001]. In the multivariate logistic analysis, stavudine treatment for > 12 months (OR, 3.67; p = 0.011) and being female (OR, 24.93; p = 0.009) are independently associated with lipoatrophy. In conclusion, the prevalence of lipoatrophy in HIV-infected Koreans receiving HAART is not uncommon. Limited use of stavudine and regular monitoring are warranted to reduce lipoatrophy.
Evaluating attendance at health education programs is important to obtain a more comprehensive evaluation of the program impact. This study investigated whether attendance at a lifestyle intervention program in a community setting would reduce risks related to metabolic syndrome. Of 545 subjects with risks related to metabolic syndrome, i.e. overweight, hypertension, dyslipidemia or diabetes, participated in this non-randomized control study, 389 subjects aged 40-71 years completed the surveys at baseline and 27 months. Intervention group (39 males and 168 females) was provided 3 individual counseling plus 28 group sessions conducted monthly on average, whereas control group (64 males and 118 females) received only 7 health information newsletters by mail. Intervention group was subcategorized into two groups according to the median attendance (87.1%): 106 subjects with high attendance (93.8 ± 4.7%) and 101 subjects with low attendance (68.6 ± 16.0%). Logistic regression analyses adjusted for age and baseline value showed that among males, the proportion with dyslipidemia risk was lower only in high attendance group compared with that in control group at 27 months [Odds ratio (OR): 0.11 (95%CI 0.02 - 0.51)] and among females, the proportion of overweight was lower only in high attendance group [OR: 0.24 (95%CI 0.07 - 0.81)]. In females, the mean total risk score calculated by adding the number of the 4 risks present decreased only in high attendance group (p < 0.001). In conclusion, high attendance at a lifestyle intervention program impacts the reduction of risks related to metabolic syndrome in a Japanese community setting.
Live kidney donation is an established form of organ donation but carries the risk of an unnecessary surgery in a normal individual for the benefit of the recipient. Despite a number of recent studies on the renal function of long-term kidney donors, little attention has been paid to the damaging effects of compensatory hyper-filtration on renal tubular cells immediately after donor nephrectomy. The present study therefore aimed to examine the immediate changes in renal function of the remaining kidney using a sheep model of unilateral nephrectomy. We used the gamma camera-based method to measure the glomerular filtration rate and the tubular excretion values after simultaneous injection of 99mTc-diethylene triamine pentaacetic acid and 131I-ortho-iodohippurate tracers. Compared were the differences in the functions between the remaining left kidney immediately after clamping the right renal pedicle and the baseline values that were measured one week before unilateral nephrectomy. After radionuclide data acquisition was completed, the right kidney was removed. The mean glomerular filtration rate (GFR) increased by 52.3% from the baseline values (29.5 ± 2.7 to 45.0 ± 6.7 ml/min; n = 40, p < 0.001), while the mean effective renal plasma flow (ERPF) increased by 40% (225.5 ± 27.8 to 357.8 ± 38.94 ml/min; p < 0.001), respectively. Mean filtration fraction was increased from 0.117 to 0.127 immediately after nephrectomy (p < 0.001). We conclude that after unilateral nephrectomy the remaining kidney immediately compensates for the loss of a donated kidney by increasing glomerular filtration rate and effective renal plasma flow.
The Wilms' tumor gene 1 (WT1) encodes a transcription factor that is involved in normal cellular development and cell survival. WT1 mRNA is overexpressed in the minimal residual disease (MRD) of patients with hematopoietic malignancy patients, particularly acute myeloid leukemia (AML). MRD represents the condition with the low levels of leukemia cells in the bone marrow and is known as a sign of recurrence. In hematopoietic malignancies, definition of remission is based on the lack of MRD at submicroscopic level. Between December 2005 and June 2008, we started to measure WT1 mRNA levels in the peripheral blood (PB) from patients by quantitative real-time PCR in Aomori Prefectural Central Hospital. Three hundreds and eight samples from 95 patients were evaluated. The patients included AML (55 patients), acute lymphoblastic leukemia (11), myelodysplastic syndrome (20), malignant lymphoma (5), chronic myeloid leukemia (1), prostatic carcinoma (1), and leukopenia (2). Among the 55 AML patients, 21 patients were pretreated with remission induction therapy. In the clinical course of 21 patients, timely therapeutic approaches could be started for relapse by the early detection of WT1 mRNA overexpression before the morphological findings were apparent. Monitoring WT1 mRNA is helpful to identify patients at high-risk relapse. High overall survival rate (71.2%, 15/21, median: 24.6 months, range 1.1-35.6 months) was achieved in 3 years. The overall survival rate of 34 post-treatment patients was 61.7% (median: 23.5 months, range 0.13-126.5 months after treatment start). In conclusion, the WT1 mRNA level is a sensitive biomarker for monitoring MRD.