A novel transfection method for mammalian cells using calcium alginate microbeads.

The direct transfer of genetic materials into mammalian cells is an indispensable technique. We have developed calcium alginate (CA) microbeads which can deliver plasmid DNAs and yeast artificial chromosomes into plant and yeast cells. In this paper, we demonstrate the effective transfection of mammalian cells by CA microbeads immobilizing plasmid DNAs.

A novel gene delivery system in plants with calcium alginate micro-beads.

We have produced micrometer-sized calcium alginate beads referred to as "bio-beads" that encapsulate plasmid DNA molecules carrying a reporter gene. In order to evaluate the efficiency of the bio-beads in mediating genetic transfection, protoplasts isolated from cultured tobacco cells (BY-2) were transfected with bio-beads containing a plasmid that carries the modified green fluorescent protein gene CaMV35S-sGFP. With the bio-beads treatment, approximately ten-fold higher GFP expression was observed after 24 h incubation compared to that with the conventional method using a naked plasmid solution.

Heel bone ultrasound predicts non-spine fracture in Japanese men and women.

A number of prospective studies in the USA and Europe have demonstrated that quantitative ultrasound (QUS) measurements predict fracture risk. To our knowledge, there has been no such study in a Japanese population, and very few studies have measured the prognostic value of QUS measurements among men, even in the USA and Europe. We performed a three-center prospective study to investigate the relationship between baseline heel QUS measurements and non-spine fracture risk.

Comparison of vertebral morphometry in the lumbar vertebrae by T1-weighted sagittal MRI and radiograph.

Purpose: In this study, we investigated the usefulness of T1-weighted sagittal MR images at the lumbar vertebrae in the vertebral morphometry, in comparison with lateral radiographs.

Subjects And Methods: The subjects were 42 men (mean age: 53.0 years) and 41 women (mean age: 57.

Pathogenesis of macular edema with branch retinal vein occlusion and intraocular levels of vascular endothelial growth factor and interleukin-6.

Purpose: To determine whether correlations between vascular endothelial growth factor (VEGF) or interleukin-6 (IL-6) contribute to the pathogenesis of macular edema in eyes of patients with branch retinal vein occlusion (BRVO).

Design: Retrospective case-control study.

Methods: Nineteen patients with macular edema with BRVO and seven patients with non-ischemic ocular disease (control group) were studied.

Development and assessment of a real-time pcr assay for rapid and sensitive detection of a novel thermotolerant bacterium, Lactobacillus thermotolerans, in chicken feces.

A new real-time PCR assay was successfully developed using a TaqMan fluorescence probe for specific detection and enumeration of a novel bacterium, Lactobacillus thermotolerans, in chicken feces. The specific primers and probe were designed based on the L. thermotolerans 16S rRNA gene sequences, and these sequences were compared to those of all available 16S rRNA genes in the GenBank database.

[Fractal analysis of trabecular architecture: with special reference to slice thickness and pixel size of the image].

Many analyses of bone microarchitecture using three-dimensional images of micro CT (microCT) have been reported recently. However, as extirpated bone is the subject of measurement on microCT, various kinds of information are not available clinically. Our aim is to evaluate usefulness of fractal dimension as an index of bone strength different from bone mineral density in in-vivo, to which microCT could not be applied.

Multi-gene gateway clone design for expression of multiple heterologous genes in living cells: conditional gene expression at near physiological levels.

Using Multisite Gateway five-DNA-fragment constructs vectors that enable expression of two tandemly situated cDNAs on a single plasmid were developed. Heterologous protein production in cells was achieved by modulating respective cDNA expression to pre-determined and different levels. Optimization of cDNA expression at near physiological protein levels was achieved using promoters from four cell cycle-dependent genes.

[Bone structure and bone quality].

Bone strength is determined by bone mineral density, geometry of bone, microarchitecture of bone and quality of bone material. Recent data have shown that the prediction of bone strength can be greatly improved by including parameters of bone structure in the analysis. However, the relative importance of bone density and architecture in the etiology of bone fractures, an issue referred to as bone quality, is poorly understood.

Identification of human T cell epitopes in Japanese cypress pollen allergen, Cha o 1, elucidates the intrinsic mechanism of cross-allergenicity between Cha o 1 and Cry j 1, the major allergen of Japanese cedar pollen, at the T cell level.

Background: Pollens from species of Cupressaceae family are one of the most important causes of respiratory allergies worldwide. In Japan, many patients with pollinosis have specific IgE to both pollens of Japanese cypress (Chamaecyparis obtusa) and Japanese cedar (Cryptomeria japonica). The sequences between Cha o 1 and Cry j 1, the major allergens of Japanese cypress and Japanese cedar pollens, respectively, are 80% identical.

Changes in angiogenic growth factor levels after gefitinib treatment in non-small cell lung cancer.

Background: To investigate the changes in angiogenic growth factor expression before and after gefitinib treatment, and the association between this expression and response to gefitinib treatment, we measured circulating levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), matrix metalloproteinase (MMP) -2 and -9, and tissue inhibitors of metalloproteinase (TIMP) -1 and -2 in patients with non-small cell lung cancer (NSCLC).

Methods: Serum and plasma samples were collected from 52 patients before and after gefitinib treatment. The levels of VEGF, bFGF, MMP-2, MMP-9, TIMP-1 and TIMP-2 were measured using a sandwich enzyme immunoassay kit.

[Increase in bone mineral density and its effect on fracture risk].

Over the past 10 years a number of clinical trials have evaluated several agents for treatment and prevention of osteoporosis, and demonstrated that these agents can increase bone mineral density (BMD) and reduce the incidence of fracture. Although the relationship between antifracture efficacy and changes in BMD varies greatly among reports, all of them suggest the presence of antifracture effect that is mediated by factors other than BMD. Fracture risk is associated with bone strength and nonskeletal risk factors such as the propensity to fall.

[Blood concentrations and secretory capacity of calcitonin in blood in osteoporosis].

The deficiency of calcitonin, CT, which has an inhibitory effect of bone resorption might be related to the pathogenesis of osteoporosis. Therefore, in this paper, in regard to the sex difference, the changes with aging, and the comparison with patients with osteoporosis and normal controls, the basal value and the secretion capacity of CT concentrations in the blood, and the metabolic clearance rate, MCR, and the production rate, PR, of CT have been reviewed.

[Bone metabolic markers and diagnosis of abnormal bone and calcium metabolism].

Bone metabolic markers increase in blood or urine, when bone formation or bone resorption accelerates. Reference values of bone metabolic markers are determined in male or female, and in pre- or post-menopause, respectively. Values of bone metabolic markers in most patients with primary osteoporosis were distributed within a reference value, mean+/-1.

[Diagnostic imaging of osteoporosis: special reference to diagnosis by radiography and DXA].

Primary osteoporosis is diagnosed by diagnostic criteria, eg. whether non-traumatic vertebral body is or not, or the severity of low bone mass. As a diagnostic imaging of osteoporosis, grading of radiographic osteopenia, and Singh index in the upper end of femur are well-known.

[Diagnosis with bone mass].

In diagnostic criteria of osteoporosis proposed by WHO, the severe osteoporosis is defined as a patient with a value for BMD or BMC more than 2.5SD below the young adult mean value in the presence of one or more fragility fractures. When severe fragility fracture occurs from the second to forth lumbar vertebra, the lumbar BMD could be over-estimated.

[Techniques for the assessment of bone microstructure].

Recent advances in non-destructive techniques for imaging of cancellous bone have made possible true three-dimensional (3D) quantification of trabecular architecture. The architectural property can be measured from 3D reconstruction of cancellous bone specimens. Examples of such procedures are model-independent assessment of trabecular dimensions, 3D connectivity estimation, degree of anisotropy and structure type assessment.

[Assessment of fracture and bone mineral density measurement].

In this paper, the assessment of radiographic vertebral fracture (prevalent fracture and incident fracture) in the diagnosis of osteoporosis and the therapeutical assessment by bone mineral density measurements are reviewed.

[Guideline for diagnosis and therapy of osteoporosis from a point of view of sex difference].

Osteoporosis occurs more common in women than that in men. In addition, there exist the sex differences in bone indexes such as bone mineral density (BMD), bone mineral content, and bone area or width, and in the decreased pattern of BMD associated with aging. Therefore, it is necessary to use a cut-off value of BMD of not women but men, as diagnostic criteria for male osteoporosis.

[Diagnosis of male osteoporosis].

Male osteoporosis is not rare because of the recent increase of elderly population. The diagnosis of male osteoporosis is based on BMD as well as in women. Although most of the cross-sectional studies have suggested that men fracture at higher BMD than women, recent prospective longitudinal studies show that the fracture risk increases as BMD decreases in men in the same way as that described in women.

[Therapeutic approaches to increased bone resorption].

The accelerated bone resorption contributes significantly to the pathophysiology of diseases such as osteoporosis, Paget's disease, hypercalcemia, bone metastasis, and inflammation of bone associated with rheumatoid arthritis. Several antiresorptive agents have been validated as a means to control bone resorption in these diseases. A recent understanding of the molecular mechanism of osteoclastic bone resorption is providing opportunities for developing novel antiresorptive agents.

[Bone mass measurement and its evaluation: their usefulness and problem].

In this paper, bone mass measurement and its evaluation have been reviewed. The bone lesions in renal osteodystrophy are classified into high- and low turnover, and the decreased rate of bone mineral density (BMD) might be different in osteitis fibrosa and adynamic bone disease. Secondary hyperparathyroid patients showed lower BMD values in mainly composed of the cortical bone than those in mainly composed of the trabecular bone.

Scalusamides A-C, new pyrrolidine alkaloids from the marine-derived fungus Penicillium citrinum.

Three new pyrrolidine alkaloids, scalusamides A-C (1-3), were isolated from the cultured broth of the fungus Penicillium citrinum, which was separated from the gastrointestine of a marine fish, and the structures were elucidated by spectroscopic data. The absolute stereochemistry of C-2 in the pyrrolidine unit was determined by HPLC analysis of a Marfey's derivative of the hydrolysate of 1, while that of 2 and 3 was assigned by comparison of spectroscopic data of 3 and reductive products of 1 and 2. On the other hand, each of 1-3 was found to be a mixture of epimers at C-7.

Three-dimensional trabecular bone architecture of the lumbar spine in bone metastasis from prostate cancer: comparison with degenerative sclerosis.

Objective: Prostate cancer frequently metastasizes to bone, inducing osteosclerotic lesions. The objective of this study was to clarify the three-dimensional (3D) trabecular bone microstructure in bone metastasis from prostate cancer by comparison with normal and degenerative sclerotic bone lesions, using microcomputed tomography (micro-CT).

Design And Materials: A total of 32 cancellous bone samples were excised from the lumbar spine of six autopsy patients: 15 metastatic samples (one patient), eight degenerative sclerotic samples (four patients) and the rest from normal sites (three patients).