Association between BIA-derived Phase Angle and Sarcopenia and Improvement in Activities of Daily Living and Dysphagia in Patients undergoing Post-Stroke Rehabilitation.

OBJECTIVES: To investigate the predictive value of the BIA-derived phase angle with respect to the functional prognosis and baseline sarcopenia in patients undergoing post-stroke rehabilitation. DESIGN: Retrospective cohort study. SETTING AND PARTICIPANTS: Overall, 577 Japanese patients admitted to a post-acute care hospital from 2016 to 2020 were recruited. MEASUREMENTS: Body composition analysis, which included BIA-derived phase angle and skeletal muscle mass, was performed using bioelectrical impedance analysis (BIA). Study outcomes included physical function assessed using the Functional Independence Measure (FIM-motor) and the level of dysphagia assessed using the Food Intake LEVEL Scale (FILS). Sarcopenia was defined as the loss of skeletal muscle mass and decreased muscle strength. Receiver operating characteristic curves were used to calculate the optimal cutoff value of BIA-derived phase angle to diagnose sarcopenia. Multivariate analyses were used to determine whether the BIA-derived phase angle at admission was associated with outcomes at discharge and baseline sarcopenia. RESULTS: After enrollment, 499 patients (mean age: 74.0 ± 13.1 years; 52.0% men) were examined. The median FIM-motor and FILS scores at admission were 47 (20-69) and 8 (7-10), respectively. Sarcopenia was observed in 43.2% of patients. After adjusting for potential confounders, BIA-derived phase angle was positively associated with FIM-motor scores at discharge (ß = 0.134, P < 0.001), FIM-motor score gain (ß = 2.504, P < 0.001), and FILS scores at discharge (ß = 0.120, P = 0.039). BIA-derived phase angle was negatively associated with the sarcopenia diagnosis at baseline (odds ratio = -0.409, P < 0.001); its cutoff value was 4.76° (sensitivity 0.800, specificity 0.790, P < 0.001) for sarcopenia diagnosis in men and 4.11° (sensitivity 0.735, specificity 0.829, P < 0.001) in women. CONCLUSION: BIA-derived phase angle was positively associated with the recovery of physical function and dysphagia level and negatively associated with baseline sarcopenia in patients undergoing post-stroke rehabilitation. The BIA-derived phase angle cutoff for sarcopenia diagnosis was 4.76° for men and 4.11° for women.

Oral Management in Rehabilitation Medicine: Oral Frailty, Oral Sarcopenia, and Hospital-Associated Oral Problems.

Oral health is a crucial but often neglected aspect of rehabilitation medicine. Approximately 71% of hospitalized rehabilitation patients and 91% of hospitalized acute care patients have impaired oral health. Poor oral condition in hospitalized patients can be attributed to factors such as age, physical dependency, cognitive decline, malnutrition, low skeletal muscle mass and strength, and multimorbidity. Another major factor is a lack of knowledge and interest in oral problems among health care workers. Recently, new concepts have been proposed, such as oral frailty, oral sarcopenia, and hospital-associated oral problems. Oral frailty, the accumulation of a slightly poor status of oral conditions and function, strongly predicts physical frailty, dysphagia, malnutrition, need for long-term care, and mortality in community-dwelling older adults. Oral sarcopenia refers to sarcopenia associated with oral conditions and function, although its definition has not yet been fully discussed. Hospital-associated oral problems are caused by disease, disease treatment, surgery, endotracheal intubation, poor self-care abilities, lack of care by medical staff, drugs, and iatrogenic factors during hospitalization. Furthermore, oral problems have negative impacts on rehabilitation outcomes, which include functional recovery, length of hospital stay, discharge home, and in-hospital mortality. Oral health management provided by dental hygienists improves not only oral status and function, swallowing function, and nutritional status but also activities of daily living, discharge home, and in-hospital mortality in post-acute rehabilitation. Oral rehabilitation, promotion, education, and medical-dental collaboration can be effective interventions for oral problems and therefore are necessary to improve rehabilitation outcomes.

Outcomes of patients receiving a massive transfusion for major trauma.

BACKGROUND: The benefits of high transfusion ratios (plasma to red blood cells and platelets to red blood cells) on survival in injured patients who receive massive transfusions remain uncertain. This study aimed to assess the association between transfusion ratios and adverse events and survival in patients undergoing massive transfusion for major trauma. METHODS: A retrospective observational study was conducted on patients who had major trauma using a Japanese national administrative database. The associations between transfusion ratios and outcomes (in-hospital mortality and incidence of adverse events) were analysed using a non-linear logistic generalized additive model (GAM). In a logistic generalized estimating equation model, adjusted for patient and hospital-level confounders, transfusion ratios were included as continuous or categorical variables (low, transfusion ratio 0·75 or less; intermediate, over 0·75 to 1·25; high, over 1·25). RESULTS: Some 1777 patients were included in the analysis, of whom 602 died in hospital. GAM plots of the transfusion ratios for in-hospital mortality demonstrated a downward convex unimodal curve. In-hospital mortality was similar with increasing transfusion ratios for plasma (adjusted odds ratio (OR) 1·13, 95 per cent c.i. 0·82 to 1·55; P = 0·446) and platelets (adjusted OR 0·84, 0·66 to 1·08; P = 0·171). Both plasma to red blood cell ratio (adjusted OR 1·77, 1·32 to 2·37; P < 0·001) and platelet to red blood cell ratio (adjusted OR 1·71, 1·35 to 2·15; P < 0·001) were significantly associated with a higher incidence of adverse events. No significant differences in in-hospital mortality were observed between the three transfusion categories (low, medium and high). CONCLUSION: In this study, transfusion strategies with high plasma to red blood cell and platelet to red blood cell ratios did not have survival benefits, but were associated with an increase in adverse events.

Efficacy and safety of ustekinumab in Japanese patients with severe atopic dermatitis: a randomized, double-blind, placebo-controlled, phase II study.

BACKGROUND: Ustekinumab, a fully human monoclonal antibody against interleukin-12/23, may potentially be effective for severe atopic dermatitis (AD) treatment. OBJECTIVES: To evaluate efficacy and safety of ustekinumab 45 mg and 90 mg in patients with severe AD. METHODS: In this randomized, placebo-controlled, phase II study, Japanese patients (aged 20-65 years) with severe or very severe AD entered a 12-week double-blind treatment period during which they received (1 : 1 : 1) ustekinumab 45 mg, 90 mg or placebo subcutaneous injections at weeks 0 and 4, with follow-up until week 24. The primary efficacy end point was percentage change from baseline in Eczema Area and Severity Index (EASI) score at week 12. Major secondary efficacy end points included the proportion of patients achieving EASI 50, EASI 75, Investigator's Global Assessment score 0-1, change from baseline Atopic Dermatitis Itch Scale and Dermatology Life Quality Index. RESULTS: A total of 79 patients were randomized [ustekinumab 45 mg (n = 24), 90 mg (n = 28), placebo (n = 27)]. Ustekinumab treatment showed nonsignificant improvement in least square mean change from baseline EASI score at week 12 [45 mg: -38·2%, 95% confidence interval (CI) -21·02-19·51; P < 0·94 and 90 mg: -39·8%, 95% CI -21·84-17·14; P < 0·81] vs. placebo (-37·5%). A nonsignificant improvement in major secondary efficacy end points was observed in both ustekinumab groups vs. placebo. The most common treatment-emergent adverse events were nasopharyngitis and worsened AD (higher in placebo vs. ustekinumab groups). CONCLUSIONS: Ustekinumab 45 mg and 90 mg did not demonstrate meaningful efficacy in Japanese patients with severe AD. The treatment was generally well tolerated.

Effectiveness of early administration of tranexamic acid in patients with severe trauma.

BACKGROUND: A reduction in mortality with the early use of tranexamic acid has been demonstrated in severely injured patients who are bleeding. However, the modest treatment effect with no reduction in blood transfusion has raised concerns. The aim of the present study was to estimate the effectiveness of regular use of tranexamic acid in severely injured patients. METHODS: This multicentre observational study used retrospectively collected data from consecutive injured patients (Injury Severity Score at least 16) treated in 15 Japanese academic institutions in 2012. A propensity score-matched analysis compared patients who did or did not receive tranexamic acid administration within 3 h of injury. Study outcomes included 28-day all-cause and cause-specific mortality, and need for blood transfusion. RESULTS: Of 796 eligible subjects, 281 were treated with tranexamic acid. Propensity score matching selected a total of 500 matched subjects (250 in each group). Tranexamic acid administration was associated with lower 28-day mortality (10·0 versus 18·4 per cent; difference -8·4 (95 per cent c.i. -14·5 to -2·3) per cent) and lower 28-day mortality from primary brain injury (6·0 versus 13·2 per cent; difference -7·2 (-12·3 to -2·1) per cent). However, there was no significant difference between groups in the need for blood transfusion (33·2 versus 34·8 per cent; difference -1·6 (-9·9 to 6·7) per cent). CONCLUSION: Early tranexamic acid use was associated with reduced mortality in severely injured patients, in particular those with a primary brain injury.

SU-E-I-115: Wavelet Analysis of Ultrasound Image for the Diagnosis of Sjögren's Syndrome.

PURPOSE: Sjögren's syndrome (SS) is an auto-immune disease presenting with dry eyes and mouth (keratoconjunctivitis sicca and xerostomia). Ultrasonography is used for the initial and non-invasive investigation of the parotid gland in the disease. The purpose of this study is to develop an image processing for diagnosis of SS by applying wavelet analysis to ultrasound image. METHODS: Ultrasound B-mode images of the parotid gland were captured and analyzed by a personal computer. A square region of interest (ROI) was set on the image and two-dimensional discrete wavelet transform was performed within the ROI. As a Result, the image was decomposed into an approximate image and three detailed images in vertical, horizontal and diagonal directions in different scales. A feature quantity for image classification was defined by calculating from the wavelet coefficients of detailed images within selected scales. The ultrasound images of 80 patients who had been referred to Nagasaki University Hospital because of suspicion of SS were analyzed. A total of 37 patients fulfilled the criteria for SS, whereas the remaining 43 patients did not. The severity of SS was graded into four degrees by sialography. The images with each feature quantity were classified by statistical cluster analysis. RESULTS: In this method, the images can be divided into two groups which mainly contained SS and non-SS. The sensitivity and specificity in the detection of SS was 78% and 95%, respectively. It was also found that the defined feature quantity tended to change with the severity of SS. CONCLUSIONS: In ultrasonography, the image analysis based on wavelet transform was useful for the diagnosis of Sjögren's syndrome.

Short communication. Development of a fine thermocouple-needle system for real-time feedback of thermal tumour ablation margin.

Thermal tumour ablation techniques such as radiofrequency (RF) ablation are applied for radical removal of local tumours as an easier, less invasive alternative to surgical resection. A serious drawback of thermal ablation, however, is that the ablation area cannot be accurately assessed during the procedure. To achieve real-time feedback and exact and safe ablation, a superfine thermocouple-needle system (TNS) comprising a 0.25-mm diameter thermocouple embedded in a 22-G, 15-cm-long needle was devised and efficacy was tested in vitro using porcine livers (n = 15) and in vivo using rabbit back muscles (n = 2) and livers (n = 3). A 17-gauge RF electrode with a 2 cm active tip was used for ablation. The TNS was inserted 1 cm from the active tip of the RF electrode and liver temperature around the electrode was measured concurrently. The RF current was cut off when the temperature reached 60°C or after 5 min at ≥50°C. Porcine livers and rabbit back muscles were then cut along a plane passing through the axes of the electrode and the TNS. In rabbit livers, contrast-enhanced CT was performed to evaluate ablation areas. Ablation areas in cut surfaces of porcine livers exhibited well-defined discoloured regions and the TNS tip precisely pinpointed the margin of the ablation area. Contrast-enhanced CT of rabbit livers showed the TNS tip accurately located at the margin of areas without contrast enhancement. These results indicate that the TNS can accurately show ablation margins and that placing the TNS tip at the intended ablation margin permits exact thermal ablation.

Effects of alfacalcidol on the contractile properties of the gastrocnemius medialis muscle in adult and old rats.

Vitamin D deficiency is associated with muscle weakness. It is unknown, however, how supra-physiological levels of vitamin D affect skeletal muscle. To investigate the effects of increased serum vitamin D (1,25 (OH)2D3 or 1,25D) levels on the contractile properties of the medial gastrocnemius muscle, adult and old female Fischer344 x Brown Norway F1 rats were orally treated with vehicle or the vitamin D analogue alfacalcidol for 1 or 6 weeks. Alfacalcidol treatment resulted in elevated 1,25D serum levels. This was accompanied by hypercalcaemia and a reduction in body mass, the latter largely attributable to a reduced food intake. However, kidney function, as reflected by normal creatinine serum levels, as well as heart mass were unaffected. The 17% reduction in maximal isometric force and power was explicable by a similar loss of muscle mass. The force-frequency relationship of the 6-week-treated old rats was shifted to the left, but neither the shape of the force-velocity relationship nor the fatigability of the muscle were altered. Supra-physiological doses of vitamin D were accompanied by significant reductions in body and muscle mass, but not by an improvement in muscle functioning. Weight loss was largely due to a reduced food intake, while the left shift in the force-frequency relation may be due to increased 1,25D levels.

Rapid detection of Acanthamoeba cysts in frozen sections of corneal scrapings with Fungiflora Y.

AIMS: To evaluate the usefulness of serial frozen sections of corneal scrapings stained with Fungiflora Y (FFY) to diagnose Acanthamoeba keratitis (AK). METHODS: Eight patients with suspected AK were studied. Serial frozen sections were made from part of the corneal epithelial scrapings and stained with FFY. The remaining corneal epithelial scrapings were submitted for laboratory culture. RESULTS: The FFY stained frozen sections were completed within an hour, and Acanthamoeba cysts were detected under a fluorescence microscope in all eight patients. The same sections were examined with a light microscope, and Acanthamoeba cysts were confirmed to be present from their morphological characteristics. Five of the eight patients had positive laboratory cultures for Acanthamoeba. CONCLUSION: FFY staining of frozen sections of corneal scrapings is a rapid and reliable technique which can be used to make an early diagnosis of AK.

Evaluation of filtering bleb function by thermography.

AIM: To determine whether thermography can be used to evaluate bleb function. METHODS: The surface temperatures of filtering blebs from 39 eyes of 33 post-trabeculectomy patients were measured using thermography. We introduced a new parameter: the temperature decrease in the filtering bleb (TDB) where TDB = (mean temperature of the temporal and nasal bulbar conjunctiva)-(temperature of the filtering bleb). The eyes were classified into good and poor intraocular pressure (IOP) control groups according to the patients' postoperative IOP. The morphological appearance of the filtering blebs were classified by slit-lamp images according to the Indiana Bleb Appearance Grading Scale. The differences in TDB were analysed statistically. RESULTS: Mean overall scores were as follows. The TDB were 0.54 (SD 0.20) degrees C and 0.21 (SD 0.18) degrees C for the good and poor IOP control groups, respectively. The difference between the TDB of the two IOP control groups was significant (p<0.0001), but there was no significant relationship between morphological classification and TDB. CONCLUSIONS: Thermographic measurements suggest that functional blebs have lower temperatures than non-functional blebs, and their morphological appearances are not affected by their temperature. Thermography may be useful for the evaluation of bleb function.

Expansion of radiofrequency ablation volume by saturated NaCl saline injection in the area of vaporization.

BACKGROUND: Vaporization around the radiofrequency (RF) electrode after RF application (RFA) limits the RF ablation area. PURPOSE: To determine whether saturated saline injected into the area of vaporization after initial RFA extends ablation area after further RFA. MATERIAL AND METHODS: RFA was performed in 18 ex vivo porcine livers and four in vivo rabbit erector spinae muscles. An RF electrode was used to ablate an area with 40W of parallel current for 15 min. The ablation margin was determined using a thermocouple, and the radius of the ablated area was measured. After RF electrode removal, saturated saline was infused through a percutaneous ethanol injection needle into the site of the original RFA in 11 liver samples and two erector spinae muscles. Three minutes later, RFA was resumed for 15 min. The remaining seven control liver samples and two spinae muscles received RFA without saline injection. The radius of the final ablated area was then measured. RESULTS: In the ex vivo study, injection of saturated saline significantly decreased tissue impedance (87.7+/-9.4 to 51.1+/-9.7 Omega, P<0.0001), and increased the mean radius of the ablated area (15.9+/-3.0 to 25.0+/-3.6 mm, P<0.0001). These significant changes were not observed without injection of saturated saline. Similar trends were found in the in vivo study. CONCLUSION: Injection of saturated saline into the area of vaporization around the RF electrode, followed by additional RFA, caused concentric expansion of the final ablation area, facilitating more efficient tumor ablation.

Oral administration of dihomo-gamma-linolenic acid prevents development of atopic dermatitis in NC/Nga mice.

Disorders of the metabolism of essential fatty acids (EFAs) are related to atopic dermatitis (AD). Concentrations of dihomo-gamma-linolenic acid (DGLA), an EFA, in the serum of AD patients are lower than those in healthy volunteers. Recently we developed a fermented DGLA oil, and examined whether oral administration of DGLA prevents development of dermatitis in NC/Nga mice, which spontaneously develop human AD-like skin lesions. NC/Nga mice were fed a diet either containing or not containing DGLA for 8 weeks under in air-uncontrolled conventional circumstances. Clinical skin severity scores were significantly lower in mice fed DGLA than in mice not fed it. Scratching behavior and plasma total IgE levels were also reduced in the DGLA group, in association with histological improvement. DGLA suppressed clinical severity of skin lesions dose-dependently, with an increase in DGLA contents in phospholipids of skin, spleen, and plasma. Discontinuation of DGLA administration resulted in the onset of dermatitis and a decrease in DGLA contents in skin, spleen, and plasma. These findings indicate that oral administration of DGLA effectively prevents the development of AD in NC/Nga mice, and that DGLA in phospholipids is a compound of key importance in the development and prevention of dermatitis.

Acute multiple brain infarction in large-artery atherosclerosis is associated with hyperhomocyst(e)inemia.

OBJECTIVES: Although hyperhomocyst(e)inemia had been proven to be a potent risk factor for ischemic stroke, it is still controversial which subtype of stroke is associated with hyperhomocyst(e)inemia. The aim of this study was to clarify the association between plasma homocyst(e)ine (Hcy) levels and stroke subtypes according to TOAST classifications based on MRI findings. METHODS: We prospectively recruited 124 consecutive first-ever ischemic stroke patients hospitalized in Toride Kyodo General Hospital. Each patient underwent brain MRI including diffusion-weighted imaging (DWI) and was evaluated for total plasma Hcy levels in addition to routine laboratory tests. RESULTS: Of the 93 patients enrolled in this study, 19 were subtyped with large-artery atherosclerosis with acute multiple brain infarction [LA-AMBI(+)], 13 with LA-AMBI(-). Patients with LA-AMBI(+) showed significantly higher plasma Hcy levels than those with LA-AMBI(-). Moreover, for LA-AMBI, the plasma Hcy level was associated with an OR of 1.3 (95% CI 1.06-1.71, P=0.017) per 1 mumol increase in concentration, and 1.4 (95% CI 1.04-1.77, P=0.025) after adjustment for age, sex and serum cholesterol levels. CONCLUSION: In the subset of patients with LA strokes, those with high Hcy levels are more likely to have AMBI than those without elevated Hcy levels possibly due to plaque instability.

Calcium supplementation does not reproduce the pharmacological efficacy of alfacalcidol for the treatment of osteoporosis in rats.

The purpose of this study was to assess whether a nutritional supply of calcium (Ca) could be substituted for alfacalcidol (ALF) administration in preventing bone loss due to estrogen deficiency. Female Wistar-Imamichi rats (8 months old) were ovariectomized (OVX) or sham-operated. OVX rats received ALF administration (0.025, 0.5, or 0.1 microg/kg, p.o., 5 times a week) with standard rodent chow [Ca 1.2%, phosphorus (P) 1.04%], a Ca-enriched diet containing 2%, 4%, or 6% Ca (Ca/P ratio of 2, 4, and 6, respectively), or a Ca/P-enriched diet (Ca/P ratio of 1.2). After 12 weeks of treatment, all rats were killed to harvest the spine, serum, and urine samples. Neither the ALF treatment nor the Ca supplement caused hypercalcemia. In the spine, ALF prevented decreases in bone mineral density (BMD) and compressive strength of lumbar spine induced by OVX. Micro-computed tomographic analysis confirmed that ALF significantly improved the trabecular bone pattern factor and the structure model index and suppressed bone destruction. In contrast, of particular interest, high-dose Ca administration did not have marked effects on bone fragility. Also, when both Ca and P were administered in high doses, BMD and mechanical strength decreased dose-dependently, urinary P excretion significantly increased, and serum parathyroid hormone level increased. Together, it is difficult to adjust the Ca supply through diet alone without disrupting the balance between serum Ca and P levels. Consequently, we conclude that ALF is beneficial for the treatment of osteoporosis, which is not achieved by the use of a Ca supplement.

Analysis of sequential immunoglobulin E-binding epitope of Japanese cedar pollen allergen (Cry j 2) in humans, monkeys and mice.

BACKGROUND: Japanese cedar (Cryptomeria japonica; CJ) pollinosis has been reported to occur naturally in Japanese monkeys (Macaca fuscata) as well as in humans. Most human patients and monkeys with pollinosis have specific IgE for Cry j 2, a major allergen of CJ pollen. OBJECTIVE: The main purpose of this study was to identify IgE B cell epitopes of Cry j 2 using a synthetic peptide in humans, monkeys and mice. METHODS: We synthesized 38 overlapping peptides that span the entire length of Cry j 2. We examined the B cell epitopes of Cry j 2 that are recognized by IgE in the sera of human patients and monkeys with pollinosis and immunized mice using synthetic peptides of Cry j 2. We also examined the reaction of Cry j 2-specific mouse monoclonal IgG antibodies to the peptides. Furthermore, we conducted a histamine release assay with leucocytes from a pollinosis patient using human serum albumin (HSA) conjugated with the peptides as a B cell epitope. RESULTS: We found that 16 of the 20 pollinosis patients who had specific IgE to Cry j 2 also exhibited IgE reaction with some Cry j 2 peptides. Of these 16 patients, 10 exhibited IgE reaction with Cry j 2 peptide no. 13 (121GQCKWVNGREICNDRDRPTA140). Five of the seven monkeys with CJ pollinosis exhibited a reaction with peptide no. 13. Furthermore, IgE in mice immunized with Cry j 2 and two mouse monoclonal IgG antibodies reacted with peptide no. 13. Peptide no. 13-conjugated HSA showed the release of histamine from basophils. Furthermore, to determine the minimum epitope in peptide no. 13, we conducted an enzyme-linked immunosorbent assay inhibition test. The core of the epitope in humans, monkeys and mice was 124KWVNGREI131. CONCLUSION: We found that 124KWVNGREI131 is an important B cell epitope recognized by IgE in humans, monkeys and mice.

Contribution of trabecular and cortical components to the mechanical properties of bone and their regulating parameters.

To evaluate the mechanical contributions of the spongiosa and cortex to the whole rat vertebra, we developed a finite element analysis (FEA) system linked to three-dimensional data from microcomputed tomography (micro-CT). Twenty-eight fifth lumbar vertebrae (L-5) were obtained from 10-month-old female rats, comprised of ovariectomized (ovx, n = 6), sham operated (n = 7), and alfacalcidol-treated after ovx (0.1 microg/kg [n = 8] and 0.2 microg/kg [n = 7]) groups. The trabecular microstructure of L-5 was measured by micro-CT. Yield strength at the tissue level (YS), defined as the value at which 0.034% of all elements reached yield stress, was calculated by the FEA. Then, the ultimate compressive load of each specimen was measured by mechanical testing. The YS of the whole bone (YSw) showed a significant correlation with ultimate load (r = 0.91, p < 0.0001). The YS values of the isolated spongiosa (YSs) and cortex (YSc) were calculated in models with varying amounts of trabecular or cortical bone mass. The mechanical contribution of the spongiosa showed a nonlinear relationship with bone mass, and ovx reduced the mean mechanical contribution of the spongiosa to the whole bone by 13% in comparison to the sham group. YSs had a strong relationship with trabecular microstructure, especially with trabecular bone pattern factor (TBPf) and structure model index (SMI), and YSc had a strong relationship with cortical bone volume. The structural parameters most strongly related to YSw were BV/TV and TBPf. Our micro-FEA system was validated to assess the mechanical properties of bone, including the individual properties of the spongiosa and cortex, in the osteoporotic rat model. We found that the mechanical property of each component had a significant relationship with the respective bone mass, volume, or structure. Although trabecular microstructure has a significant relationship with bone strength, in ovx bone with deteriorated trabecular microstructure, the strength depended mainly on the cortical component.

A comparison of alfacalcidol and menatetrenone for the treatment of bone loss in an ovariectomized rat model of osteoporosis.

We conducted this study to evaluate the characteristic effects of alfacalcidol (ALF) and menatetrenone (VK) in preventing bone loss using an ovariectomized rat model of osteoporosis. Bilateral ovariectomy (OVX) or sham operation was performed on 10-month-old female Wistar rats. OVX caused a significant decrease in the bone mass and the mechanical strength of the lumbar vertebra as well as the femur 6 months after surgery. VK treatment (30 mg/kg, food intake) required a 6-month period to prevent the bone loss induced by estrogen deficiency, whereas ALF (0.1 or 0.2 mg/kg, p.o.) increased the bone mass and the mechanical strength of the lumbar vertebra as well as the femur in a 3-month treatment period, far above the level in the sham-operated rats. Neither ALF or VK caused hypercalcemia, despite administration for as long as 6 months. By doing a micro-CT analysis of the vertebral trabecular microstructure, it was revealed that ALF treatment increased the interconnections and the plate-like structures and that VK significantly increased the trabecular number. It was also indicated that the increase in spinal strength by ALF treatment was closely associated with improvement of the microstructure, but not VK. The results of histomorphometric analysis showed that ALF caused a significant suppression of bone resorption yet maintained formation in the endocortical perimeter, and also stimulated bone formation in the periosteal perimeter, thereby causing an increase in cortical area. No marked effect of VK on histomorphometric parameters was observed, whereas VK as well as ALF maintained the material strength at femoral midshaft of the normal level, suggesting that VK affected bone quality and thereby prevented the decrease in mechanical strength of femur caused by OVX. In conclusion, it was demonstrated that the two drugs, ALF and VK, differed markedly in their potency and mechanisms for improving bone strength. These results have important implications in understanding the characteristic actions of vitamin K and active vitamin D on bone metabolism.

Seasonal changes of humoral and cellular immune responses to Japanese cedar (Cryptomeria japonica) pollen allergens in Japanese monkeys (Macaca fuscata) with pollinosis.

The natural occurrence of Japanese cedar [Cryptomeria japonica (CJ)] pollinosis has been reported in Japanese monkeys (Macaca fuscata). The present study was designed to investigate seasonal changes in immunological reactions to CJ pollen allergens in monkeys with CJ pollinosis. Blood samples were collected from six monkeys with CJ pollinosis before and after CJ pollen season. Seasonal changes in specific IgE and IgG to major allergens (Cry j 1 and Cry j 2) were observed before and after CJ pollen season. The humoral responses decreased significantly before CJ pollen and increased after CJ pollen season. Similar seasonal changes in peripheral blood mononuclear cells proliferative responses to CJ allergens were observed before and after CJ pollen season. These humoral and cellular immune responses might serve as a biomarker for assessing new immunotherapies for monkeys with pollinosis.

Preclinical evaluation of an immunotherapeutic peptide comprising 7 T-cell determinants of Cry j 1 and Cry j 2, the major Japanese cedar pollen allergens.

BACKGROUND: Peptide immunotherapy is a new approach to treating allergic diseases, but a therapeutic peptide for Japanese cedar pollinosis has not yet been developed. OBJECTIVE: The aim of this study is to prepare and preclinically evaluate a hybrid peptide comprising 7 T-cell determinants of Cry j 1 and Cry j 2, the major Japanese cedar pollen allergens. METHODS: The recombinant hybrid peptide was prepared after immunodominance of 7 T-cell determinants was confirmed by means of PBMC proliferation assay in 113 volunteers with pollinosis. The hybrid peptide was compared with a mixture of the 7 T-cell determinants in a dose-dependent PBMC proliferation assay in 6 volunteers with pollinosis. PBMC proliferation and binding activity of serum IgE antibody against the hybrid peptide, Cry j 1, and Cry j 2 were investigated in 48 volunteers with pollinosis. RESULTS: The hybrid peptide induced T-cell proliferation with an average 100-fold lower concentration than a mixture of the 7 peptides. PBMCs from 44 (92%) of 48 volunteers proliferated against the hybrid peptide, with significant correlation (r = 0.87) in T-cell proliferation against Cry j 1 and Cry j 2. No serum IgE antibodies specific to Cry j 1 or Cry j 2 bound to the hybrid peptide. CONCLUSION: A hybrid peptide comprising 7 T-cell determinants has the potential for inducing T-cell proliferative responses that is superior to the potential of a mixture of the T-cell determinants and comparable with that of Cry j 1 and Cry j 2. The hybrid peptide will be of use in specific immunotherapy against Japanese cedar pollinosis.

Effect of trabecular bone contour on ultimate strength of lumbar vertebra after bilateral ovariectomy in rats.

To test the hypothesis that the effect of trabecular microarchitecture on bone strength varies with the duration of estrogen loss, we evaluated the relationship between three-dimensional (3D) parameters for trabecular microarchitecture and bone minerals with the compressive load of the lumbar vertebra in rats. Female Sprague-Dawley rats (n = 190) were divided into 19 groups. Ten rats were killed at day 0. Half of the remaining rats underwent bilateral ovariectomy (ovx), and the others were subjected to sham surgery. Ten rats from each group were killed at 3, 7, 11, 14, 28, 42, 56, 70, and 84 days postsurgery. Urinary deoxypyridinoline and serum osteocalcin increased significantly in the ovx group from days 28 and 11, respectively, compared with the sham group. Bone mineral content (BMC) and bone mineral density (BMD) of the fifth lumbar body diminished from days 42 and 84, respectively, compared with the sham group. In ovx rats, trabecular bone volume (BV/TV), measured using 3D images of microcomputed tomography, diminished from day 28 compared with both baseline control and sham. The trabecular bone pattern factor (TBPf) and structure model index (SMI) increased from day 28 in the ovx group compared with both baseline control and sham. Ultimate compression loads diminished at day 28 compared with baseline control and decreased progressively thereafter. Neither of these parameters changed in the sham group during the same period. Within 4 weeks post-ovx, TBPf, SMI, and BV/TV correlated with load (p < 0.01). BMC and BMD correlated with load from 6 weeks post-ovx (p < 0.01). Stepwise regression analysis showed that TBPf was the most significant determinant of load within 4 weeks post-ovx (coefficient of determination [R(2)] = 0.669; p < 0.01). SMI correlated with TBPf (R(2) = 0.968; p < 0.01). Moreover, R(2) for ultimate load indicated higher values of 0.975 with TBPf and SMI. However, BMC was the most significant determinant of load from 6 weeks post-ovx (R(2) = 0.511; p < 0.01), as it was in the sham group. These data suggest that changes in trabecular bone contour with increased bone turnover are critical for reducing lumbar bone strength during the early post-ovx period in rats.