Multiplex SNP genotyping in whole blood using an integrated microfluidic lab-on-a-chip.

Pharmacogenetics has often been touted as a cornerstone for precision medicine as detailed knowledge of a specific genetic makeup may allow for accurate predictions of a patient's individual drug response. Still, the widespread use of genetic tests is limited as they remain expensive and cumbersome, requiring sophisticated tools and highly trained personnel. In order for pharmacogenetics to reach its full potential, more cost-effective and easily accessible genotyping methods are desired. To meet these challenges, we present a silicon-based integrated microsystem for the detection of multiple single nucleotide polymorphisms (SNPs) directly from human blood. The device combines a blood lysis chamber, a cross-flow filter, a T-junction mixer, and a microreactor for quantitative polymerase chain reaction (qPCR). Using this device, successful on-chip genotyping of two clinically relevant SNPs in human CYP2C9 gene was demonstrated with TaqMan assays, starting from blood. The two SNPs were detected simultaneously by introducing a sequence of plugs, each containing a different set of primers and probes. The method can be easily extended to detect several SNPs. The microsystem described here offers a rapid, reproducible, and accurate sample-to-answer technology enabling multiplex SNP profiling in point-of-care settings, bringing pharmacogenetics-based precision medicine a step closer to reality.

Bioenergetics and the role of soluble cytochromes C for alkaline adaptation in gram-negative alkaliphilic Pseudomonas.

Very few studies have been conducted on alkaline adaptation of Gram-negative alkaliphiles. The reversed difference of H(+) concentration across the membrane will make energy production considerably difficult for Gram-negative as well as Gram-positive bacteria. Cells of the alkaliphilic Gram-negative bacterium Pseudomonas alcaliphila AL15-21(T) grown at pH 10 under low-aeration intensity have a soluble cytochrome c content that is 3.6-fold higher than that of the cells grown at pH 7 under high-aeration intensity. Cytochrome c-552 content was higher (64% in all soluble cytochromes c) than those of cytochrome c-554 and cytochrome c-551. In the cytochrome c-552-dificient mutant grown at pH 10 under low-aeration intensity showed a marked decrease in µ max⁡ [h(-1)] (40%) and maximum cell turbidity (25%) relative to those of the wild type. Considering the high electron-retaining abilities of the three soluble cytochromes c, the deteriorations in the growth of the cytochrome c-552-deficient mutant could be caused by the soluble cytochromes c acting as electron storages in the periplasmic space of the bacterium. These electron-retaining cytochromes c may play a role as electron and H(+) condenser, which facilitate terminal oxidation at high pH under air-limited conditions, which is difficult to respire owing to less oxygen and less H(+).

Laser measurement of H- ions in a field-effect-transistor based radio frequency ion source.

Hydrogen negative ion density measurements are required to clarify the characteristics of negative ion production and ion source performance. Both of laser photodetachment and cavity ring down (CRD) measurements have been implemented to a field-effect-transistor based radio-frequency ion source. The density ratio of negative hydrogen ions to electrons was successfully measured by laser photodetachment and effect of magnetic filter field on negative ion density was confirmed. The calculated CRD signal showed that CRD mirrors with >99.990% reflectivity are required and loss of reflectivity due to cesium contamination should be minimized.

H- beam extraction from a cesium seeded field effect transistor based radio frequency negative hydrogen ion source.

H(-) beam was successfully extracted from a cesium seeded ion source operated using a field effect transistor inverter power supply as a radio frequency (RF) wave source. High density hydrogen plasma more than 10(19) m(-3) was obtained using an external type antenna with RF frequency of lower than 0.5 MHz. The source was isolated by an isolation transformer and H(-) ion beam was extracted from a single aperture. Acceleration current and extraction current increased with the increase of extraction voltage. Addition of a small amount of cesium vapor into the source enhanced the currents.

Long-term results of conventional varus half-wedge proximal femoral osteotomy for the treatment of osteonecrosis of the femoral head.

We have previously described the mid- to long-term results of conventional simple varus intertrochanteric osteotomy for osteonecrosis of the femoral head, showing that 19 of the 26 hips had good or excellent results. We extended the follow-up to a mean of 18.1 years (10.5 to 26) including a total of 34 hips in 28 patients, with a mean age at surgery of 33 years (19 to 53). There were 18 men and ten women and 25 hips (74%) had a satisfactory result with a Harris hip score ≥ 80. In all, six hips needed total hip replacement (THR) or hemiarthroplasty. The collapse of the femoral head or narrowing of the joint space was found to have progressed in nine hips (26%). Leg shortening after osteotomy was a mean of 19 mm (8 to 36). With conversion to THR or hemiarthroplasty as the endpoint, the ten-year survival rate was 88.2% (95% confidence interval (CI) 82.7 to 93.7) and the 20-year survival rate was 79.7% (95% CI 72.1 to 87.3); four hips were converted at ten years and other two hips were converted at 20 years. Shortening of the leg after osteotomy remains a concern; however, the conventional varus half-wedge osteotomy provides favourable long-term results in hips with less than two-thirds of the medial part of the femoral head affected by necrotic bone and with normal bone superolaterally.

The Chiari pelvic osteotomy for patients with dysplastic hips and poor joint congruency: long-term follow-up.

We report the mid- to long-term (mean 20.3 years, 10 to 32.5) results of the Chiari pelvic osteotomy in patients with pre- to advanced stage osteoarthritis in dysplastic hips. We followed 163 Japanese patients (173 hips) with a mean age at surgery of 20 years (9 to 54). Overall, 124 hips (72%) had satisfactory results, with Harris hip scores ≥ 80. Satisfactory results were seen in 105 of 134 hips with pre- or early osteoarthritis (78%) and 19 of 39 hips with advanced osteoarthritis (49%). A total of 15 hips (9%) underwent a total hip replacement (THR) with a mean interval between osteotomy and THR of 16.4 years. With conversion to THR as the endpoint, the 30-year survival rate was 85.9% (95% confidence interval 82.3 to 89.5). It was 91.8% for patients with pre- or early osteoarthritis and 43.6% for those with advanced osteoarthritis (p < 0.001). We now perform the Chiari osteotomy for patients with dysplastic hips showing poor joint congruency and who prefer a joint-conserving procedure to THR.

Radio frequency ion source operated with field effect transistor based radio frequency system.

Characteristics of radio frequency (RF) plasma production are investigated using a field effect transistor inverter power supply as an RF wave source. With the frequency of around 0.3 MHz, an electron density over 10(18) m(-3) is produced in argon plasma. Although lower densities are obtained in hydrogen plasma, it drastically increased up to 5x10(18) m(-3) with an axial magnetic field of around 100 G applied in the driver region. Effects of the magnetic field and gas pressure are investigated in the RF produced plasma with the frequency of several hundred kilohertz.

Flattening of sagittal spinal curvature as a predictor of vertebral fracture.

UNLABELLED: In a 12-year prospective study of 100 adult volunteers, incident vertebral fractures were analyzed for potential predictors. Decreased sagittal spinal curvature and pre-existing fractures were identified as independent predictors. The risk of vertebral fracture increased to 8.6-fold with a 1 SD decrease in both thoracic and lumbar curves. INTRODUCTION: Physiological spinal curvature acts as a shock absorber: however, it has not been demonstrated whether reduced spinal curvature is a risk of vertebral fracture. The aim of this prospective study is to assess the relationship between the extent of sagittal spinal curvature and incidence of vertebral fractures. METHODS: One hundred community-based volunteers aged 50+ years were followed radiologically for at least 10 years. Entire spine radiograms of volunteers in erect position were used for evaluating sagittal spinal curvatures, grade of osteoporosis, and incident vertebral fractures. RESULTS: Mean baseline age was 61.9 years and mean follow-up period was 12.0 years. In multivariate analysis, preexisting vertebral fracture (relative risk [RR] 3.70, 95% confidence interval [CI] 1.22-11.20), and a decrease in either thoracic or lumbar curvature by 1-SD (RR 3.06, 95% CI 1.04-9.00) were independent predictors. The fracture risk even increased in the presence of a 1-SD decrease in both thoracic and lumbar curvature (RR 8.64, 95% CI 1.39-53.78). CONCLUSIONS: Reduction of physiological curvatures in both thoracic and lumbar spine led to the increased risk of vertebral fracture by more than eightfold. The importance of evaluating sagittal spinal alignment should be emphasized, because it might indicate the risk of the first vertebral fracture, which is another strong predictor of subsequent fractures.

Health-related quality of life in patients with systemic lupus erythematosus after medium to long-term follow-up of hip arthroplasty.

Health-related quality of life (HRQOL) of systemic lupus erythematosus (SLE) patients with hip arthroplasty after medium to long-term follow-up has not been reported. We conducted a retrospective study for SLE patients with osteonecrosis of the femoral head (ONF). Forty-seven consecutive arthroplasties were performed in 36 patients. Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Medical Outcome Survey Short Form 36 (SF-36) and Harris hip score were evaluated. Two patients died before the four-year follow-up and two patients were lost to follow-up. The remaining 43 hips in 32 patients with an average age at surgery of 35 years and an average follow-up of 12.0 years (range 4.0-25.0) were assessed. Bipolar hemiarthroplasty was performed for 18 hips in 12 patients, and total hip arthroplasty (THA) was performed for 25 hips in 20 patients. The mean WOMAC scores for pain and function at the recent followup were 90.8 +/- 8.5points and 79.0 +/- 18.3 points. Patients with THA had significantly high scores in SF-36 physical functioning (P < 0.05) and bodily pain (P < 0.03) compared to those with bipolar hemiarthroplasty. Although improvement could not reach the level of general population, the hip arthroplasty contributed to support HRQOL of SLE patients.

An advantage for concavities in shape perception by chimpanzees (Pan troglodytes).

The significance of concavity in object shape perception by chimpanzees (Pan troglodytes) was investigated in a matching-to-sample procedure. For the task, chimpanzees were required to choose a polygon stimulus that was identical in shape to a sample. The incorrect alternative was defined by the addition or subtraction of a concave or convex apex. Chimpanzees were more sensitive to the concave deformation than to the convex deformation. This tendency conforms to the theories of human visual perception that have treated concave features as important factors in reconstructing three-dimensional structures from two-dimensional images. Our results suggest that shape representation in chimpanzees is similar to that in humans and that chimpanzees visually process two-dimensional images in the same manner as humans.

Three-dimensional computer-aided design based design sensitivity analysis and shape optimization of the stem using adaptive p-method.

The number of stem designs for total hip arthroplasty is increasing, and occasionally design changes have yielded unexpected clinical results. At present, we are not able to clearly identify which parameter of the stem is most important, and the optimum value of many parameters. The goals of this study were to identify which parameter is most important, to understand the effect of design change, and to find the optimum stem shape. For this purpose, we used adaptive p-method together with three-dimensional computer-aided design software program for the design sensitivity analysis (DSA) and shape optimization of the stem. The results suggested that increasing the lateral and medial width of the distal cross-section together with decreasing the medial-lateral width and the medial radius of the distal cross-section from the default value would lead to a decrease in the largest maximum principal stress of the distal cement. The medial width of middle cross-section, however, was not so simple. The result of DSA suggested that decreasing this parameter from the default value decreased the stress in the distal cement, but the optimum shape was obtained by increasing this parameter. The method used in this study will assist our engineers and surgeons in the process of modifying and optimizing the stem design.

Pre-coated femoral components in hybrid total hip arthroplasty. Results at 11 years.

We present the medium-term results of hybrid total hip arthroplasties using pre-coated stems with a second-generation cementing technique. The 128 hips in 111 patients (18 men and 93 women) were followed up at a mean of 11 years after surgery. The mean age at the time of surgery was 61 years. Both components of one hip were removed at ten months after surgery for infection. None of the other 127 femoral components showed possible, probable, or definite loosening at the most recent follow-up. Five acetabular components were revised for aseptic loosening, recurrent dislocation, or displacement of the polyethylene liner from the metal shell. The mean Harris hip score at follow-up was 84 points. A pre-coated femoral component with a second-generation cementing technique provides good clinical function and survival in the medium term.

Coverage dependent desorption dynamics of deuterium on Si(100) surfaces: interpretation with a diffusion-promoted desorption model.

We studied coverage dependence of time-of-flight (TOF) spectra of D2 molecules thermally desorbed from the D/Si(100) surface. The mean translational energies Et of desorbed D2 molecules were found to increase from 0.20+/-0.05 eV to 0.40+/-0.04 eV as the desorption coverage window was decreased from 1.0 ML> or =thetaD> or =0.9 ML to 0.2 ML> or =thetaD> or =0 ML, being consistent with the kinetics switch predicted in the interdimer mechanism. The measured TOF spectra were deconvoluted into 2H, 3H, and 4H components by a curve fitting method along the principle of detailed balance. As a result, it turned out that the desorption kinetics changes from the 4H to the 3H situation at high coverage above thetaD=0.9 ML, while the 2H desorption is dominant for a quite wide coverage region up to thetaD=0.8 ML. A dynamic desorption mechanism by which the desorption is promoted by D-atom diffusion to dangling bonds was proposed.

Mid-term prognosis of non-traumatic osteonecrosis of the femoral head.

We followed, prospectively, 77 patients (90 hips) with early-stage, non-traumatic osteonecrosis of the femoral head for a mean period of nine years. At the time of final review, 56 (62%) were symptomatic. Using the Cox model, the initial radiological stage, the progression of staging, the reduction in size of the lesions and the percentage of necrotic volume on MRI were identified as risk factors. Using the Harris score of < 70 or surgery as the endpoint, the cumulative rates of survival were 60.0% at one year, 43.3% at two years, 38.9% at five years and 37.2% at ten years. Survival curves demonstrated that clinical deterioration could occur 90 months after the initial diagnosis, suggesting that asymptomatic patients should be followed carefully for several years. Radiological time-dependent reduction in size without progressive collapse may represent repair even when the collapse is minimal.

Comparison of the surgical approaches for a Chiari pelvic osteotomy.

We present the mid- to long-term results of the Chiari pelvic osteotomy for dysplastic hips. We followed 135 hips in 129 patients, with a mean age at the time of surgery of 24 years, for a mean of 16.2 years We used the anterior iliofemoral approach without trochanteric osteotomy in the initial 31 hips. Thereafter, we used transtrochanteric approaches in an attempt to ensure that the osteotomy was at the most appropriate level, and to advance the high-riding greater trochanter distally. The next 79 hips therefore underwent a posterolateral approach and the most recent 25 hips an Ollier lateral U approach. The clinical result was excellent or good in 103 hips (77%). The outcome in 104 hips in which we used a transtrochanteric approach was superior, the osteotomy level was more appropriate and a Trendelenburg gait less common than in 31 hips in which we used an anterior approach. We therefore recommend the use of a transtrochanteric approach in order to ensure that the osteotomy is at an appropriate level and in order to achieve effective distal advancement of the high-riding greater trochanter.

Strength of bond to bone and cytotoxicity of sintered bodies of hydroxyapatite/zirconia composite particles.

Regarding sintered bodies of hydroxyapatite (HAP)/zirconia (PSZ) composite particles prepared by covering the surface of HAP particles with PSZ particles, their strength of the bond to bone after implanting them into an organism and their cytotoxicity were evaluated. Cytotoxicity tests were conducted by the colony formation method. Cytotoxicity was not observed in the sintered bodies of HAP/PSZ composite particles, or in sintered monolithic HAP body and sintered monolithic PSZ body. The strength of bond between a sintered body and a bone was evaluated by measuring the shear strength at the interface between them after a fixed period following implantation of a sintered body into a rabbit femur. In all cases of the sintered monolithic HAP body, and the sintered bodies of HAP/PSZ composite particles and monolithic PSZ body, when the implantation period increased, the shear strength of the sintered body/bone interface tended to increase. In particular, this tendency was very high in the case of sintered bodies of HAP/PSZ with weight ratios of 1/1.0 and 1/1.5, the shear strength for each reaching 20 MPa 12 months after implantation.

Comparative biochemical studies of carotenoids in catfishes.

The carotenoids of 12 species of Siluriformes fishes (eight families) were investigated from a comparative biochemical point of view. The patterns of carotenoids in catfishes belonging to the family Siluridae were quite different from those of the other seven families of catfishes (Bagridae, Amblycipitidae, Clariidae, Plotosidae, Ictaluridae, Callichthyidae and Malapteruridae). 7, 8-Dihydro-beta-carotene; 7, 8, 7', 8'- and 7, 8, 9, 10-tetrahydro-beta-carotene; (3R)-7', 8'-dihydro-beta-cryptoxanthin; 7, 8-dihydrolutein A; 7, 8-dihydrolutein B; parasiloxanthin; 7', 8'-dihydroparasiloxanthin; and 4 or 4'-hydroxyparasiloxanthin were characteristic carotenoids found in only one family, Siluridae, and these carotenoids accounted for 24-60% of total carotenoids. In catfishes belonging to the other seven families except Siluridae, the carotenoid patterns were very similar and the most predominant carotenoid was zeaxanthins (23-56%).

Cytomegalovirus DNA among children attending two day-care centers in Tokyo.

BACKGROUND: In the USA, a high prevalence rate of cytomegalovirus (CMV) excretion among children in day-care centres was reported. However, there is no research about the prevalence rate of CMV among children in day-care centres in Japan. METHODS: The CMV excretion was studied in 54 children's saliva samples, collected from two different day-care centres in Tokyo. As a control, the prevalence of CMV was studied among 61 healthy children who did not attend any day-care centers. The CMV DNA in saliva were examined by polymerase chain reaction (PCR) analysis with one pair of primers for the immediate early region. The sequence of CMV genomes were examined in CMV PCR positive samples. RESULTS: Of the 54 saliva samples, 20.6% (6/29) and 24% (6/25) were CMV PCR positive in children at A and B day-care centres, respectively. The overall positivity of CMV PCR in saliva was 22.2% (12/54). Of the 61 saliva samples as the control study, 6.5% (4/61) were CMV PCR positive. There was a difference in the positivity in each age group of day-care centres and normal control. Each sample of the same day-care center gave conclusive and identical sequence results. CONCLUSION: We suspected that in each day-care center that there was one prevailing viral strain. We suppose that CMV infections were acquired inside the day-care centres. This is a first report which described viral transmission in day-care centres in Japan.

The sphericity of the bearing surface in total hip arthroplasty.

This study evaluated the sphericity of bearing surfaces in total hip arthroplasty. The out-of-roundness of metal femoral heads, the inner surface of polyethylene liners, and commercially available ball bearings was measured. The hip prostheses were obtained directly from the manufacturers. The sphericity of the bearing surfaces was significantly inferior to that of the ball bearings. The sphericity of the femoral head on the sagittal plane was inferior to that on the transverse plane. Several significant differences were found among different manufacturers. The sphericity of the femoral head on the sagittal plane and that of polyethylene significantly improved in 1999 and 2000 compared with those in 1995. Further improvement is desirable, however, because good sphericity is expected to prolong the functional performance of the prosthesis after total hip arthroplasty.