Discrimination of Frailty Phenotype by KinectTM-Based Stepping Parameters.

Background: Frailty increases the risk of falling, hospitalization, and premature death, necessitating practical early-detection tools. Objectives: To examine the discriminative ability of KinectTM-based stepping parameters for identifying frailty phenotype. Design: Population-based cross-sectional study. Setting: Eighteen neighborhoods near Tokyo Metropolitan Institute for Geriatrics and Gerontology, Itabashi, Tokyo, Japan. Participants: In total, 563 community-dwelling older adults aged ≥75 years without mobility limitations, neurological disease, or dementia were included. Measurements: Step number (SN) and knee total movement distance (KMD) during a 20-s stepping test were evaluated using the KinectTM infrared depth sensor. Results: The number (%) of participants with frailty were 51 (9.1). The area under the receiver operating characteristic curves (95% confidence interval) of a parameter consisting of SN and KMD for frailty was 0.72 (0.64, 0.79). Conclusions: Stepping parameters evaluated using KinectTM provided acceptable ability in identifying frailty phenotype, making it a practical screening tool in primary care and home settings.

Assessment of effect of partial sterility on mating performance in sweetpotato weevil (Coleoptera: Curculionidae).

The sterile insect technique (SIT) is widely used to suppress or eradicate target pest insect populations. Although the effectiveness of SIT depends on the ability of released sterile males to mate with and inseminate wild females, the use of gamma radiation to induce sterility negatively impacts reproductive cells as well as somatic cells. Consequently, sterilization by irradiation drastically diminishes mating performance over time. In the current study, we evaluated the effect of irradiation dose intensity on fertility, mating propensity, and mating competitiveness in sweetpotato weevil, Cylas formicarius elegantulus (Summers) (Coleoptera: Curculionidae), for 16 d after irradiation. Although the mating propensity of males irradiated with 200 Gy, the dose currently used to induce complete sterility of C. f. elegantulus in the SIT program in Okinawa Prefecture, was equal to that of nonirradiated weevils for the first 6 d, the mating propensity of males irradiated with doses between of 75 and 150 Gy was maintained for the first 12 d. The potential fertilization ability of weevils was highly depressed compared with the control weevils, even in those treated with 75 Gy. Mating performance was severely compromised in weevils that were irradiated with a dose of 100 Gy or more. These results demonstrate that partial sterilization can be highly advantageous in eradication programs for the sweetpotato weevil. We discuss the advantages of the application of partial irradiation in insect eradication programs.

Sphingosine kinase expression increases intracellular sphingosine-1-phosphate and promotes cell growth and survival.

Sphingosine-1-phosphate (SPP) is a bioactive lipid that has recently been identified as the ligand for the EDG family of G protein-coupled cell surface receptors. However, the mitogenic and survival effects of exogenous SPP may not correlate with binding to cell-surface receptors (Van Brocklyn, J.R., M.J. Lee, R. Menzeleev, A. Olivera, L. Edsall, O. Cuvillier, D.M. Thomas, P.J.P. Coopman, S. Thangada, T. Hla, and S. Spiegel. 1998. J. Cell Biol. 142:229-240). The recent cloning of sphingosine kinase, a unique lipid kinase responsible for the formation of SPP, has provided a new tool to investigate the role of intracellular SPP. Expression of sphingosine kinase markedly increased SPP levels in NIH 3T3 fibroblasts and HEK293 cells, but no detectable secretion of SPP into the medium was observed. The increased sphingosine kinase activity in NIH 3T3 fibroblasts was sufficient to promote growth in low- serum media, expedite the G(1)/S transition, and increase DNA synthesis and the proportion of cells in the S phase of the cell cycle with a concomitant increase in cell numbers. Transient or stable overexpression of sphingosine kinase in NIH 3T3 fibroblasts or HEK293 cells protected against apoptosis induced by serum deprivation or ceramide elevation. N,N-Dimethylsphingosine, a competitive inhibitor of sphingosine kinase, blocked the effects of sphingosine kinase overexpression on cell proliferation and suppression of apoptosis. In contrast, pertussis toxin did not abrogate these biological responses. In Jurkat T cells, overexpression of sphingosine kinase also suppressed serum deprivation- and ceramide-induced apoptosis and, to a lesser extent, Fas-induced apoptosis, which correlated with inhibition of DEVDase activity, as well as inhibition of the executionary caspase-3. Taken together with ample evidence showing that growth and survival factors activate sphingosine kinase, our results indicate that SPP functions as a second messenger important for growth and survival of cells. Hence, SPP belongs to a novel class of lipid mediators that can function inside and outside cells.

Development of Simple, Objective Chair-Standing Assessment of Physical Function in Older Individuals Using a KinectTM Sensor.

BACKGROUND: With increasing interest in addressing quality of life of older individuals, tests such as the Functional Independence Measure (FIM) are widely used measures of infirmity and burden of care. However, these scales are largely qualitative and especially problematic when assessing movement-based tasks. While effective, reliable analysis of human movement is technically complicated and expensive; an infrared depth sensor is potentially a low-cost, portable devise which may provide a quantitative aspect to clinical testing. OBJECTIVE: to assess the utility of the KinectTM sensor in providing an objective evaluation of human movement using an oft measured ADL (chair stand). DESIGN: Cross-sectional study. SETTING: Community, geriatric day-care center in Japan. PARTICIPANTS: Men (n=136) and women (n=266) between 50 and 93 years of age, consisting of healthy (HE; n=312) and physically frail (FR; n= 90) individuals. MEASUREMENTS: Subjects completed two trials of the chair stand, conducted without assistance. Trials were timed and recorded with KinectTM v2. Coronal plane angle (CPA) was determined by a line transecting the shoulder-center and waist relative to the vertical axis and was used to assess quality of the chair stand movement pattern. RESULTS: Age, height, and body mass were not different between groups. CPA was significantly greater in FR (29.3 ± 8.3°) than HE (19.5 ± 6.5°). CPA and age were significantly related (r=0.148, p<0.01). An optimal threshold for CPA identifying frailty was determined by a receiver-operator characteristic curve with a CPA of 23.1° providing the greatest combination of sensitivity (79%) and specificity (73%). CONCLUSION: During the chair stand, frail older adults adopted a forward lean position (increased CPA) compared to healthy older adults. This compensatory posture appears to facilitate torso rotation while reducing lower-limb muscular effort during standing. As such, CPA serves as an indicator of reduced lower-body function in older, frail adults.

Effect of irradiation on mating ability in the male sweetpotato weevil (Coleoptera: Curculionidae).

The sterile insect technique (SIT) is widely used for suppressing or eradicating target pest insect populations. The effectiveness of SIT depends on the ability of released sterile males to mate with and inseminate wild females. Irradiation is the effective manner to sterilize mass-reared insects. The negative impacts of this procedure are not limited to damage on reproductive cells. Gamma-radiation damages the epithelial tissue of midgut, which affects the alimentation in insects. Irradiated males alter their mating behavior over time because of the depression of metabolic activity by sterilization. In this study, we evaluated the male mating performance and sexually compatibility of irradiated male Cylas formicarius elegantulus (Summers) (Coleoptera: Curculionidae) with a 200-Gy dose, as currently used in the SIT program in Okinawa Prefecture, throughout 16 d after irradiation in the laboratory. The mating ability of irradiated males did not differ from that of control males for about a week. However, the mating ability of irradiated male drastically decreased thereafter. We consider that irradiated male C. formicarius elegantulus with a 200-Gy dose had no major effect on male mating behavior approximately for a week after irradiation.

Overcoming MDR-associated chemoresistance in HL-60 acute myeloid leukemia cells by targeting sphingosine kinase-1.

We examined the involvement of sphingosine kinase-1, a critical regulator of the sphingolipid balance, in susceptibility to antineoplastic agents of either sensitive or multidrug-resistant acute myeloid leukemia cells. Contrary to parental HL-60 cells, doxorubicin and etoposide failed to trigger apoptosis in chemoresistant HL-60/Doxo and HL-60NP16 cells overexpressing MRP1 and MDR1, respectively. Chemosensitive HL-60 cells displayed sphingosine kinase-1 inhibition coupled with ceramide generation. In contrast, chemoresistant HL-60/ Doxo and HL-60/VP16 had sustained sphingosine kinase-1 activity and did not produce ceramide during treatment. Enforced expression of sphingosine kinase-1 in chemosensitive HL-60 cells resulted in marked inhibition of apoptosis that was mediated by blockade of mitochondrial cytochrome c efflux hence suggesting a control of apoptosis at the pre-mitochondrial level. Incubation with cell-permeable ceramide of chemoresistant cells led to a sphingosine kinase-1 inhibition and apoptosis both prevented by sphingosine kinase-1 over-expression. Furthermore, F-12509a, a new sphingosine kinase inhibitor, led to ceramide accumulation, decrease in sphingosine 1-phosphate content and caused apoptosis equally in chemosensitive and chemoresistant cell lines that is inhibited by adding sphingosine 1-phosphate or overexpressing sphingosine kinase-1. F-12509a induced classical apoptosis hallmarks namely nuclear fragmentation, caspase-3 cleavage as well as downregulation of antiapoptotic XIAP, and release of cytochrome c and SMAC/Diablo.

Telomerase activity in human endometrium.

Human uterine endometrium undergoes a complex pattern of changes in proliferation and secretory activity during the menstrual cycle. In the present study, telomerase activity in normal endometrium was examined using a non-radioisotope PCR-based telomeric repeat amplification protocol assay. Various levels of telomerase activity were detected in the 60 normal endometrial samples examined, depending on the phase of the menstrual cycle. Of 21 proliferative-phase endometrial samples, 20 (95%) expressed telomerase activity, whereas 8 of 19 (42%) secretory-phase or menstrual endometrial samples did (P = 0.002). Five of nine (56%) samples from atrophic endometrium from postmenopausal women also expressed telomerase activity. Eleven of 21 (52%) endometrial samples in the proliferative phase expressed high telomerase activity detectable after 100-fold dilution of extracts, whereas none of the 19 endometrial samples from the secretory phase or during menstruation and none of the 9 postmenopausal endometrial samples did (P < 0.001). The highest activity was observed in the late proliferative phase, but activity dramatically decreased with the progression of the secretory phase. Surprisingly, the levels of telomerase activity detected in the late proliferative phase were comparable to those detected in the endometrial cancers examined. Immunohistochemical analysis of the expression of proliferating cell nuclear antigen revealed that telomerase activity is closely correlated with endometrial cell proliferative activity. These findings indicate that normal endometrium expresses telomerase, the activity of which changes dramatically over the course of the menstrual cycle, suggesting in turn that telomerase is a regulated enzyme linked to cellular proliferation and that hormone functions may be involved in its regulation.

A burst-promoting activity derived from the human bone marrow stromal cell line KM-102 is identical to the granulocyte-macrophage colony-stimulating factor.

Recently, several human bone marrow stromal cell lines have established and produced hematopoietic growth factors. One of these factors, a burst-promoting activity (BPA), was purified from 6 liters of serum-free conditioned medium cultured from stromal cell line KM-102, which was stimulated by phorbol myristate acetate (PMA) and calcium ionophore A23187. This stimulation induced 60 times more production of BPA than the unstimulated control culture. BPA was purified 4000-fold by sequential fractionation using ammonium sulfate precipitation, anion-exchange and lentil lectin affinity chromatographies, high performance gel filtration chromatography, and reversed phase high performance liquid chromatography. Purified BPA gave a single broad band of protein with a molecular weight of approximately 18 kd, as assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The concentration required for half maximal growth of early erythroid colonies was estimated as 10 pg/ml or 0.6 pM. At a higher concentration (125 pg/ml) this factor also stimulates the growth of granulocyte, macrophage, and eosinophil colonies in agar culture. The profile of amino acid composition is very similar to that of the human granulocyte-macrophage colony-stimulating factor (GM-CSF) deduced from its complementary DNA sequence. The result of amino-terminal sequence analysis strongly suggests that the purified material consists of GM-CSF and tetrapeptide-deleted GM-CSF. Moreover, antibody against GM-CSF completely neutralized the biological activities of this factor. These results indicate that the human bone marrow stromal cell line secretes GM-CSF as a burst-promoting activity and GM-CSF may play a significant role in the interaction between stem cells and stromal cells in the hematopoietic microenvironment.

Functional characterization of human sphingosine kinase-1.

Sphingosine kinase catalyzes the phosphorylation of sphingosine to form sphingosine 1-phosphate (SPP), a novel lipid mediator with both intra- and extracellular functions. Based on sequence identity to murine sphingosine kinase (mSPHK1a), we cloned and characterized the first human sphingosine kinase (hSPHK1). The open reading frame of hSPHK1 encodes a 384 amino acid protein with 85% identity and 92% similarity to mSPHK1a at the amino acid level. Similar to mSPHK1a, when HEK293 cells were transfected with hSPHK1, there were marked increases in sphingosine kinase activity resulting in elevated SPP levels. hSPHK1 also specifically phosphorylated D-erythro-sphingosine and to a lesser extent sphinganine, but not other lipids, such as D,L-threo-dihydrosphingosine, N, N-dimethylsphingosine, diacylglycerol, ceramide, or phosphatidylinositol. Northern analysis revealed that hSPHK1 was widely expressed with highest levels in adult liver, kidney, heart and skeletal muscle. Thus, hSPHK1 belongs to a highly conserved unique lipid kinase family that regulates diverse biological functions.

The effect of leustroducsin B on the production of cytokines by human mesenchymal cells.

Leustroducsin B (LSN-B), a novel colony-stimulating factor (CSF) inducer, has been shown to have various biologic activities in vivo. To compare the CSF-inducing activity of LSN-B in vitro with that of the well-known cytokine inducer, interleukin-1beta (IL-1beta), bacterial lipopolysaccharide (LPS), and phorbol 12-myristate 13-acetate (PMA), we measured granulocyte (G)-CSF and granulocyte-macrophage (GM)-CSF levels that were induced with the stimuli in several mesenchymal cells. The results indicated that each stimulant displayed a different profile in the induction of G-CSF and GM-CSF. Next, to investigate if LSN-B induces cytokines other than G-CSF and GM-CSF, we characterized cytokines that were induced with LSN-B from bone marrow stromal cells (BMSCs). The results showed that a variety of cytokines, including G-CSF and GM-CSF, were induced in both clonal and primary BMSCs. From these results, we speculate that LSN-B induces cytokine production via a regulatory pathway distinct from that of IL-1beta, LPS, or PMA and that this signaling of LSN-B might lead to the production of a variety of cytokines in BMSCs. In addition, from our in vitro and in vivo results, we speculate that the biologic activities of LSN-B in vivo might be based on its own cytokine-inducing activity even though the target cell type of LSN-B in vivo remains to be determined.

Antideuteron production in 158A GeV/c Pb + Pb collisions

The invariant cross section as a function of transverse momentum for antideuterons produced in 158A GeV/c per nucleon Pb+Pb central collisions has been measured by the NA44 experiment at CERN. This measurement, together with a measurement of antiprotons, allows for the determination of the antideuteron coalescence parameter. The extracted coalescence radius is found to agree with the deuteron coalescence radius and radii determined from two particle correlations.

Effect of N-ethylmaleimide on Ca-inhibition of Physarum myosin.

We have established a quick method for preparing Physarum myosins whose actin-activated ATPase activities are inhibited by microM levels of Ca2+ (from plasmodial stage: Kohama, K. & Kendrick-Jones, J. (1986) J. Biochem. 99, 1433-1446; and from amoebal stage: Kohama, K., Takano-Ohmuro, H., Tanaka, T., Yamaguchi, Y., & Kohama, T. (1986) J. Biol. Chem. 261, 8022-8027). N-Ethylmaleimide alkylates sulfhydryl (SH) groups on the heavy chains in the heads of the plasmodial myosin. The actin-activated ATPase activity of the modified myosin was significantly decreased when assayed in low Ca2+ concentrations. Moreover, the activity remained low even when the Ca2+ concentrations was increased, i.e., the myosin was desensitized. For complete desensitization, about 4 mol SH per mol myosin (500,000 Mr) must be modified. These residues are probably the "reactive thiols" which have been predicted from primary structure studies to be conserved among myosins of higher and lower eukaryotes. Ultraviolet absorption spectra of the modified and intact myosins showed a peak at 277 nm. The height of this peak in intact myosin was reduced when the Ca2+ concentration was increased. This Ca-induced reduction was hardly detectable in the modified myosin although Ca-binding activity to myosin did not appear to be affected by the modification. We interprete these results that Ca2+ may change the conformation of the myosin heavy chain by binding to myosin and speculate that impairment of this process upon modification could cause the desensitization to Ca2+ in the ATPase activity.

Sphingosine-1-phosphate in cell growth and cell death.

Recent evidence suggests that branching pathways of sphingolipid metabolism may mediate either apoptotic or mitogenic responses depending on the cell type and the nature of the stimulus. While ceramide has been shown to be an important regulatory component of apoptosis induced by tumor necrosis factor alpha and Fas ligand, sphingosine-1-phosphate (SPP), a further metabolite of ceramide, has been implicated as a second messenger in cellular proliferation and survival induced by platelet-derived growth factor, nerve growth factor, and serum. SPP protects cells from apoptosis resulting from elevations of ceramide. Inflammatory cytokines stimulate sphingomyelinase, but not ceramidase, leading to accumulation of ceramide, whereas growth signals also leading to accumulation of ceramide, whereas growth signals also stimulate ceramidase and sphingosine kinase leading to increased SPP levels. We propose that the dynamic balance between levels of sphingolipid metabolites, ceramide, and SPP, and consequent regulation of different family members of mitogen-activated protein kinases (JNK versus ERK), is an important factor that determines whether a cell survives or dies.

Large uterine myoma with erythropoietin messenger RNA and erythrocytosis.

BACKGROUND: Myomatous erythrocytosis syndrome (erythrocytosis associated with a uterine myoma) has multiple proposed etiologies, one of which is altered erythropoietin production. CASE: A 28-year-old woman, gravida 0, para 0, presented with a solitary, degenerated uterine myoma that was 34-36 weeks' gestational size and erythrocytosis. After GnRH agonist treatment and myomectomy, the tumor was analyzed by reverse transcription-polymerase chain reaction. Specific erythropoietin primer with erythropoietin messenger RNA was noted. CONCLUSION: Erythropoietin production by myomata might cause erythrocytosis in myomatous erythrocytosis syndrome.

A technique of minilaparotomy-assisted vaginal hysterectomy.

In minilaparotomy-assisted vaginal hysterectomy, the operation begins vaginally by opening the peritoneal folds and ligating the uterine vessels and uterosacral ligaments. Minilaparotomy is then performed for myomectomy, cutting the fallopian tubes and the utero-ovarian ligaments and detaching any adhesions on the anterior peritoneal angle. In 26 women who underwent this procedure, the feasibility rate was 100% and no intraoperative complications or postoperative morbidity was observed (except in one case of ovarian bleeding), indicating that vaginal hysterectomy assisted by minilaparotomy is a feasible approach for hysterectomy in the setting of large myomas, myomas with adhesions caused by endometriosis or previous pelvic surgery, and adenomyosis.

Parameters for plaque formation in the potency assay of Japanese measles vaccines.

Parameters for plaque formation by measles vaccine strains licensed in Japan were studied. For the plaque test, inoculum volume was one of the critical factors for obtaining an appropriate titre of the sample. A linear relationship between the inoculum volume and the apparent reciprocal titre was discovered, enabling the comparison of absolute titres. Another factor affecting the infectivity was the strain-specific temperature sensitivity in the plaque assay. Although all the vaccine strains tested showed the highest titre at 35 degrees C, the pattern of the temperature sensitivity differed from one strain to another. These factors must be taken into consideration in order to obtain an appropriate titre of a vaccine virus.

Quantitative analysis of serum alpha 1-acid glycoprotein levels in normal and diabetic pregnancy.

In an attempt to clarify the mechanism of lipid metabolism during pregnancy, alpha 1-acid glycoprotein (alpha 1-AG) was analyzed in normal and diabetic pregnant women. Seventy-two determinations of serum alpha 1-AG levels were performed in 18 diabetic pregnant women and 82 determinations in 82 normal pregnant women in all three trimesters and within 14 days postpartum. Serum alpha 1-AG levels in both normal and diabetic pregnant women decreased throughout pregnancy and rapidly increased postpartum. In all gestational stages, the serum alpha 1-AG levels were lower in diabetic women than in normal women, but the differences were not significant. No significant correlation was obtained between serum alpha 1-AG and hemoglobin A1 (HbA1) in diabetic patients. On the contrary, the serum triglyceride levels increased during pregnancy and decreased postpartum in both groups of subjects. These findings suggest that serum alpha 1-AG plays an important role in the activation of lipoprotein lipase during pregnancy.

Characterization of B-5354c, a new sphingosine kinase inhibitor, produced by a marine bacterium.

B-5354c is a new inhibitor of sphingosine kinase from a novel marine bacterium, SANK 71896. Kinetic study revealed that B-5354c inhibits sphingosine kinase with a Ki value of 12/microM. The inhibition is noncompetitive with respect to sphingosine. The compound also inhibits sphingosine-1-phosphate formation in human platelets. Experiments using synthetic derivatives of B-5354c indicate that all the three functional groups, i.e., the long unsaturated aliphatic chain, 4-amino and 3-hydroxyl groups are necessary to inhibit sphingosine kinase.

F-12509A, a new sphingosine kinase inhibitor, produced by a discomycete.

In the course of our screening for inhibitors of sphingosine kinase, we found an active compound from a culture broth of a discomycete, Trichopezizella barbata SANK 25395. The structure of the compound, named F-12509A, was elucidated by a combination of spectroscopic analyses, to be a new sesquiterpene quinone consisting of a drimane moiety and a dihydroxybenzoquinone. Enzyme kinetic analyses showed that F-12509A inhibits sphingosine kinase activity in a competitive manner with respect to sphingosine, with a Ki value of 18 microM.

Novel microbial metabolites of the phoslactomycins family induce production of colony-stimulating factors by bone marrow stromal cells. II. Isolation, physico-chemical properties and structure determination.

Leustroducsins (LSNs) A, B and C, novel inducers of colony-stimulating factors (CSFs), were isolated from culture broth of Streptomyces platensis SANK 60191 mainly by ethyl acetate extraction and preparative reverse-phase HPLC. The molecular weights and molecular formulae of LSNs A, B and C are 641: C32H52O10NP, 669: C34H56O10NP and 669: C34H56O10NP, respectively. The structure elucidation revealed that they belong to the phoslactomycin group antibiotics, and their structures contain an alpha,beta-unsaturated delta-lactone, an amino group, a phosphate ester and a cyclohexane ring moiety. The structures differ only at the substituent bound to the cyclohexane ring.