Ethnic differences in cross-sectional associations between impaired glucose regulation, identified by oral glucose tolerance test or HbA1c values, and cardiovascular disease in a cohort of European and South Asian origin.

AIMS: We contrasted impaired glucose regulation (prediabetes) prevalence, defined according to oral glucose tolerance test or HbA1c values, and studied cross-sectional associations between prediabetes and subclinical/clinical cardiovascular disease (CVD) in a cohort of European and South Asian origin. METHODS: For 682 European and 520 South Asian men and women, aged 58-85 years, glycaemic status was determined by oral glucose tolerance test or HbA1c thresholds. Questionnaires, record review, coronary artery calcification scores and cerebral magnetic resonance imaging established clinical plus subclinical coronary heart and cerebrovascular disease. RESULTS: Prediabetes was more prevalent in South Asian participants when defined by HbA1c rather than by oral glucose tolerance test criteria. Accounting for age, sex, smoking, systolic blood pressure, triglycerides and waist-hip ratio, prediabetes was associated with coronary heart disease and cerebrovascular disease in European participants, most obviously when defined by HbA1c rather than by oral glucose tolerance test [odds ratios for HbA1c -defined prediabetes 1.60 (95% CI 1.07, 2.39) for coronary heart disease and 1.57 (95% CI 1.00, 2.51) for cerebrovascular disease]. By contrast, non-significant associations were present between oral glucose tolerance test-defined prediabetes only and coronary heart disease [odds ratio 1.41 (95% CI 0.84, 2.36)] and HbA1c -defined prediabetes only and cerebrovascular disease [odds ratio 1.39 (95% CI 0.69, 2.78)] in South Asian participants. Prediabetes defined by HbA1c or oral glucose tolerance test criteria was associated with cardiovascular disease (defined as coronary heart and/or cerebrovascular disease) in Europeans [odds ratio 1.95 (95% CI 1.31, 2.91) for HbA1c prediabetes criteria] but not in South Asian participants [odds ratio 1.00 (95% CI 0.62, 2.66); ethnicity interaction P = 0.04]. CONCLUSIONS: Prediabetes appeared to be less associated with cardiovascular disease in the South Asian than in the European group. These findings have implications for screening, and early cardiovascular prevention strategies in South Asian populations.

The role of FoxC1 in early Xenopus development.

FoxC1 is an important transcription factor in vertebrate development since its mutation in humans results in Axenfeld-Rieger syndrome. In the mouse, disturbance of its function causes congenital hydrocephalus and abnormalities in the development of various mesodermal derivatives. In this report, we provide one mechanistic basis for the requirement for FoxC1 in vertebrate development. We find that, in Xenopus laevis embryos, FoxC1 expression is regulated by the maternal T-box transcription factor VegT, via the nodal sub-family of TGFbeta signaling transducers. We show that at the late neurula to early tailbud stage, FoxC1 depletion causes the down-regulation of adhesion molecules, EP and E cadherin, as well as members of the Ephrin/EphR signaling families in the mesoderm germ layer resulting in the loss of adhesion and apoptosis of mesodermal cells.

Clinical evaluation of expanded input dynamic range in Nucleus cochlear implants.

OBJECTIVE: The aim of this study was to investigate whether expanded instantaneous input dynamic ranges (IIDRs) in the Nucleus cochlear implant system benefit speech perception in the laboratory and listening in the real world. DESIGN: Until recently, Nucleus cochlear implants have used an IIDR of approximately 30 dB. In this study, an IIDR of 31 dB was compared with 46 dB and 56 dB in the SPEAR3 research processor with nine adult implant recipients. Subjects were given two, 2-wk blocks of take-home experience with each of the three IIDRs. A single IIDR setting was used in each trial period. During the take-home experience with the expanded IIDRs, subjects used two programs: a standard program (with clinically measured electrode dynamic ranges) and a program with adjusted thresholds (decreased T levels). After each block of take-home experience, speech perception testing was conducted for CNC words in quiet (at 45 dB and 55 dB SPL) and for CUNY sentences in the presence of multi-taker babble. RESULTS: On average, CNC word recognition at low presentation levels was significantly better with the 46 dB and 56 dB IIDRs, compared with the 31 dB IIDR; however, there was no significant difference between the 46 dB and 56 dB IIDR conditions. These benefits were greater for standard programs than for reduced T level programs. For CUNY sentences in babble, group results indicated no significant difference in performance across IIDR. The three IIDRs were rated similarly in real-life listening situations, and two of the subjects expressed tolerance problems with the expanded standard IIDRs. CONCLUSIONS: IIDRs of 46 and 56 dB provided benefit in accessing low-level speech without a decrement in sentence perception in babble. Most subjects accepted the standard, wider IIDR programs in everyday life. No significant differences were found between the 46 dB and 56 dB IIDR programs.

Control of a hand grasp neuroprosthesis using an electroencephalogram-triggered switch: demonstration of improvements in performance using wavepacket analysis.

Volitionally modulated electroencephalographic (EEG) waves were monitored for the purpose of controlling a hand neuroprosthesis in people with tetraplegia. The region of the EEG signal spectrum monitored was the occipital alpha wave (8-13 Hz), and volitional modulation was achieved with the opening and closing of the eyes. In a set of 13 trials evaluated, a subject with tetraplegia successfully completed ten trials undertaking stimulated grasp and release using the EEG-triggered switch. EEG signal data recorded during the 13 trials were also post-processed off-line using wavepacket analysis. Following this signal processing, the speed and reliability of the EEG-triggered switch, when operated by the subject with tetraplegia, was significantly improved (p < 0.002). Such improvements provide system performance that is likely to be acceptable to a neuroprosthesis user during activities of daily life.

Synergistic control of stimulated pronosupination with the stimulated grasp of persons with tetraplegia.

The control of stimulated forearm pronosupination in concert with stimulated hand grasp of persons with tetraplegia has been investigated. It has been shown that hand grasp stability increased as supination was achieved. In accordance with this, a strategy of object acquisition has been proposed incorporating pronosupination and hand grasp. It has been proposed that, after object acquisition in the pronated posture, that supination be used to increase grasp stability. Three types of pronosupination control which act in synchrony with grasp were implemented incorporating this principle. The three types used were position-controlled pronator stimulation, touch-controlled pronator stimulation, and constant pronation stimulation. These controllers played a supporting role to the separate user control of hand grasp and release. The three controllers were evaluated and compared using a standardized test procedure that incorporated stimulated pronosupination control with stimulated grasp. Such methods of pronosupination control are likely to provide enhanced options for improving upper extremity function using electrical stimulation.

Timing of endogenous activin-like signals and regional specification of the Xenopus embryo.

Signaling by activin-like ligands is important for induction and patterning of mesoderm and endoderm. We have used an antibody that specifically recognizes the phosphorylated and activated form of Smad2, an intracellular transducer of activin-like ligands, to examine how this signaling pathway patterns the early mesendoderm. In contrast to the simple expectation that activin-like signaling should be highest on the dorsal side of the gastrula stage embryo, we have found that while Smad2 phosphorylation is highest dorsally before gastrulation, signaling is attenuated dorsally and is highest on the ventral side by mid-gastrulation. Early dorsal initiation of Smad2 phosphorylation results from cooperation between the vegetally localized maternal transcription factor VegT and dorsally localized beta-catenin. The subsequent ventral appearance of Smad2 phosphorylation is dependent on VegT, but not on signaling from the dorsal side. Dorsal attenuation of Smad2 phosphorylation during gastrulation is mediated by early dorsal expression of feedback inhibitors of activin-like signals. In addition to regulation of Smad2 phosphorylation by the expression of activin-like ligands and their antagonists, the responsiveness of embryonic cells to activin-like ligands is also temporally regulated. Ectopic Vg1, Xnr1 and derrière all fail to activate Smad2 phosphorylation until after the midblastula transition, and the onset of responsiveness to these ligands is independent of transcription. Furthermore, the timing of cellular responsiveness differs for Xnr1 and derrière, and these distinct temporal patterns of responsiveness can be correlated with their distinctive phenotypic effects. These observations suggest that the timing of endogenous activin-like signaling is a determinant of patterning in the early Xenopus embryo.

The role of maternal axin in patterning the Xenopus embryo.

Regulation of the stability of beta catenin protein is a critical role of Wnt signaling cascades. In early Xenopus development, dorsal axis specification depends on regulation of beta catenin by both cytoplasmic and nuclear mechanisms. While the cytoplasmic protein axin is known as a key component of the cytoplasmic beta catenin degradation complex, loss-of-function studies are needed to establish whether it is required for dorso-ventral patterning in the embryo, and to test where in the embryo it carries out its function. Here, we show that embryos lacking maternal axin protein have increased levels of soluble beta catenin protein and increased nuclear localization of beta catenin in ventral nuclei at the blastula stage. These embryos gastrulate abnormally and develop with excessive notochord and head structures, and reduced tail and ventral components. They show increased expression of dorsal markers, including siamois, Xnr3, chordin, gsc, Xhex, and Otx2, decreased expression of Xwnt 8 and Xbra, and little alteration of BMP4 and Xvent1 and -2 mRNA levels. The ventral halves of axin-depleted embryos at the gastrula stage have dramatically increased levels of chordin expression, and severely decreased levels of Xwnt 8 mRNA expression, while BMP4 transcript levels are only slightly reduced. This dorso-anterior phenotype is rescued by axin mRNA injected into the vegetal pole of axin-depleted oocytes before fertilization. Interestingly, the phenotype was rescued by ventral but not dorsal injection of axin mRNA, at the 4-cell stage, although dorsal injection into wild-type embryos does cause ventralization. These results show directly that the localized ventral activity of maternal axin is critical for the correct patterning of the early Xenopus embryo.

Early events in the mammalian germ line.

Germ cells represent the genetic and cellular link between generations, as well as the transmitters of inherited diseases. Despite their central importance, not much is known about the molecular mechanisms whereby a germ cell lineage becomes set aside during development, or how the germ cells, once formed, migrate to the gonads and combine with somatic cells to make a gonad. This article provides a brief review of current knowledge on these issues, with particular focus on the mammalian germ line.

Maternal VegT is the initiator of a molecular network specifying endoderm in Xenopus laevis.

During cleavage stages, maternal VegT mRNA and protein are localized to the Xenopus embryo's vegetal region from which the endoderm will arise and where several zygotic gene transcripts will be localized. Previous loss-of-function experiments on this T-box transcription factor suggested a role for VegT in Xenopus endoderm formation. Here, we test whether VegT is required to initiate endoderm formation using a loss of function approach. We find that the endodermal genes, Bix1, Bix3, Bix4, Milk (Bix2), Mix.1, Mix.2, Mixer, Xsox17 alpha, Gata4, Gata5, Gata6 and endodermin, as well as the anterior endodermal genes Xhex and cerberus, and the organizer specific gene, Xlim1, are downstream of maternal VegT. We also find that the TGF beta s, Xnr1, Xnr2, Xnr4 and derrière rescue expression of these genes, supporting the idea that cell interactions are critical for proper endoderm formation. Additionally, inhibitory forms of Xnr2 and Derrière blocked the ability of VegT mRNA injection to rescue VegT-depleted embryos. Furthermore, a subset of endodermal genes was rescued in VegT-depleted vegetal masses by induction from an uninjected vegetal mass. Finally, we begin to establish a gene hierarchy downstream of VegT by testing the ability of Mixer and Gata5 to rescue the expression of other endodermal genes. These results identify VegT as the maternal regulator of endoderm initiation and illustrate the complexity of zygotic pathways activated by VegT in the embryo's vegetal region.

Vegetal localization of maternal mRNAs is disrupted by VegT depletion.

VegT is an essential maternal regulator of germ layer specification in Xenopus. The localization of VegT mRNA to the vegetal cortex of the oocyte during oogenesis ensures its inheritance by vegetal and not animal cells, and directs the differentiation of vegetal cells into endoderm. Similarly localized mRNAs, Vg1 and Bicaudal-C, are also inherited by vegetal cells, while germ plasm-associated mRNAs, such as Xcat2, become incorporated into vegetally derived primordial germ cells. Although mRNA localization is clearly important for tissue specification, the mechanism of mRNA anchoring to the oocyte vegetal cortex is not understood. Here, we examine the role of VegT in cortical localization. We report that depletion of VegT mRNA caused the release of Vg1 mRNA from the vegetal cortex and a reduction of Vg1 protein, without affecting the total amount of Vg1 transcript. Furthermore, we found that Bicaudal-C and Wnt11 mRNAs were also dispersed, but not degraded, by VegT depletion, while the localization of Xcat2 and Xotx1 mRNAs was unaffected. This effect was specific to the loss of VegT mRNA and not VegT protein, since a morpholino oligo against VegT, that blocked translation without degrading mRNA, did not disperse the vegetally localized mRNAs. Therefore, a subset of localized mRNAs is dependent on VegT mRNA for anchoring to the vegetal cortex, indicating a novel function for maternal VegT mRNA.

Detection of fatigue in the isometric electrical activation of paralyzed hand muscles of persons with tetraplegia.

Paralyzed muscle fatigue is the eventual depression of force due to either prolonged or repetitive electrical stimulation of motor units. The robustness and safety of future functional electrical stimulation (FES) systems will rely on their ability to detect the onset of muscle fatigue. The relative degree of muscle activation can be estimated by monitoring the M-wave. The aim of this study was to test a proposed method of quantitative fatigue assessment that detects muscle force output and its corresponding M-wave measured concurrently. The detection of force and M-wave concurrently allows any reduction in muscle force output to be attributed to either changes in the fatigue state of the stimulated muscle or changes in the degree of stimulus activation of that muscle. The fatigue assessment scheme can thereby accommodate the corresponding changes in muscle force caused by an alteration in the stimulation intensity during fatigue. The Extensor Digitorum Communis (EDC), Extensor Pollicis Longus (EPL), and Flexor Pollicis Longus (FPL) muscles of two C5/C6 tetraplegic men were studied. Stimulation recruitment tests over the pulsewidth range from 0 to 200 micros, were performed at intervals during 20 min of maximal stimulation (200 micro/s). Muscle force correlated to the M-wave parameter, second phase area, with mean correlation coefficients of greater than 0.82, when the muscle was in either a nonfatigued or fatiguing state. After the initial force, likely to be primarily due to the fast glycolytic (FG) motor units, had declined the M-wave demonstrated only minor changes throughout the fatigue of muscle force during 20 min of constant maximal stimulation. The second phase area and root-mean-square (rms) of the M-wave [see Fig. 2(a) reflected muscle activation during modulated stimulation and also remained relatively constant during the fatigue-related force decline when the muscle was stimulated at a constant intensity. This detection of M-wave parameters satisfies the defined requirement for a myoelectric parameter that indicates electrical activation, but is relatively invariant to muscular fatigue. Index Terms-Electrical stimulation, electromyography (EMG), functional electrical stimulation (FES), muscle fatigue, spinal cord injury, tetraplegia.

The use of bilateral stimulated hand grasp in the bimanual activities of persons with tetraplegia.

Grasp and release has been provided to both upper extremities of subjects with tetraplegia using percutaneous and fully implanted stimulation. This is to determine quantitatively the performance of these subjects in three bimanual tests using two handed stimulation. Their performance here is compared with two separate cases: the same tests using single handed stimulation assisted by a nonstimulated hand and using two unstimulated hands. It was found that bilateral stimulation significantly improved the efficacy of performing the bimanual tests assessed over that using unilateral stimulation assisted by the nonstimulated hand (in two of three tests) or no stimulation three of three tests). These results show quantitative evidence of the benefits of providing bilateral stimulation for the completion of bimanual tasks for persons with tetraplegia who are appropriate candidates for stimulated grasp. These benefits were observed especially in cases where the ability of the subject in completing the task using unilateral stimulation was not strong.

Quantitative evaluation of two methods of control of bilateral stimulated hand grasps in persons with tetraplegia.

Electrical stimulation has been applied to the paralyzed muscles of both hands of two persons with tetraplegia using percutaneous and implantable electrodes. Two separate methods of user control were being investigated. The first monitored the myoelectric signals from the user's own sternocleidomastoid muscles and the second monitored wrist joint angle. These signals were used as commands to modify the stimulated grasps. The hands were instrumented to detect the degree of hand closure and grip force and the users matched these to specific target parameters using the controller during tracking tasks. Performance in these tracking tasks was measured quantitatively. Wrist control was found to be less sensitive to the direction of hand opening/closing required than the myoelectric control. The user's performance with the myoelectric control demonstrated sensitivity to the target size and the speed of hand movement in response to the command control. The wrist controller required less training than the myoelectric controller for users to become proficient in its use. Based on these results, the wrist controller and the myoelectric controller both provide successful control of bilateral hand grasp and release. Of the two controllers, the wrist controller is likely to provide the greater ease of use, although it is only available to the population of users with active wrist extension.

Beta-catenin signaling activity dissected in the early Xenopus embryo: a novel antisense approach.

Xenopus embryos develop dorsal/ventral and anterior/posterior axes as a result of the activity of a maternal Xwnt pathway, in which beta-catenin is an essential component, acting as a transactivator of transcription of zygotic genes. However, the questions of where and when beta-catenin is required in early embryogenesis have not been addressed directly, because no loss-of-function method has been available. Here we report the use of a novel antisense approach that allows us to target depletion of protein to individual blastomeres. When a "morpholino" oligo complementary to beta-catenin mRNA is injected into early embryos, it depletes beta-catenin protein effectively through the neurula stage. By targeting the oligo to different cleavage blastomeres, we block beta-catenin activity in different areas and at different times. Dorsal vegetal injection at the 2- and 4-cell stages blocks dorsal axis formation and at the 8-cell stage blocks head formation, while A-tier injection at the 32-cell stage causes abnormal cement gland formation. This approach shows the complex involvement of Xwnt pathways in embryonic patterning and offers a rapid method for the functional analysis of both maternal and early zygotic gene products in Xenopus.

The putative wnt receptor Xenopus frizzled-7 functions upstream of beta-catenin in vertebrate dorsoventral mesoderm patterning.

We have isolated one member of the frizzled family of wnt receptors from Xenopus (Xfz7) to study the role of cell-cell communication in the establishment of the vertebrate axis. We demonstrate that this maternally encoded protein specifically synergizes with wnt proteins in ectopic axis induction. Embryos derived from oocytes depleted of maternal Xfz7 RNA by antisense oligonucleotide injection are deficient in dorsoanterior structures. Xfz7-depleted embryos are deficient in dorsal but not ventral mesoderm due to the reduced expression of the wnt target genes siamois, Xnr3 and goosecoid. These signaling defects can be restored by the addition of beta-catenin but not Xwnt8b. Xfz7 thus functions upstream of the known GSK-3/axin/beta-catenin intracellular signaling complex in vertebrate dorsoventral mesoderm specification.

The onset of germ cell migration in the mouse embryo.

Mouse primordial germ cells (PGCs) are specified between embryonic day 6.5 (E6.5) and E7.5, when they have been visualized as an alkaline phosphatase-positive (AP+) cell population in the developing allantois. By E8.5, they are embedded in the hind-gut epithelium. Previous experiments have suggested different sites for PGCs' origin, and it is unclear how they reach the gut epithelium. We have used transgenic mice expressing GFP under a truncated Oct4 promoter to visualize living PGCs. We find GFP+/AP+ cells in the posterior end of the primitive streak as a dispersed population of cells actively migrating into the allantois, and directly into the adjacent embryonic endoderm. Time-lapse analysis shows these cells to be actively migratory from the time they exit the primitive streak.

The role of cadherins during primordial germ cell migration and early gonad formation in the mouse.

Primordial germ cells (PGCs) are the founder cells of the gametes. In mammals, PGCs migrate from the hindgut to the genital ridges, where they coalesce with each other and with somatic cells to form the primary sex cords. We show here that, in both sexes, PGCs express P- and E-cadherins during and after migration, and N-cadherin at post-migratory stages. E-Cadherin is not expressed by PGCs whilst in the hindgut, but is upregulated as they leave. Blocking antibodies against E-, but not P-cadherin cause defective PGC-PGC coalescence, and in some cases, ectopic PGCs.

Mesoderm induction in Xenopus is a zygotic event regulated by maternal VegT via TGFbeta growth factors.

The maternal transcription factor VegT is important for establishing the primary germ layers in Xenopus. In previous work, we showed that the vegetal masses of embryos lacking maternal VegT do not produce mesoderm-inducing signals and that mesoderm formation in these embryos occurred ectopically, from the vegetal area rather than the equatorial zone of the blastula. Here we have increased the efficiency of the depletion of maternal VegT mRNA and have studied the effects on mesoderm formation. We find that maternal VegT is required for the formation of 90% of mesodermal tissue, as measured by the expression of mesodermal markers MyoD, cardiac actin, Xbra, Xwnt8 and alphaT4 globin. Furthermore, the transcription of FGFs and TGFbetas, Xnr1, Xnr2, Xnr4 and derrière does not occur in VegT-depleted embryos. We test whether these growth factors may be endogenous factors in mesoderm induction, by studying their ability to rescue the phenotype of VegT-depleted embryos, when their expression is restricted to the vegetal mass. We find that Xnr1, Xnr2, Xnr4 and derrière mRNA all rescue mesoderm formation, as well as the formation of blastopores and the wild-type body axis. Derrière rescues trunk and tail while nr1, nr2 and nr4 rescue head, trunk and tail. We conclude that mesoderm induction in Xenopus depends on a maternal transcription factor regulating these zygotic growth factors.

Expression of the Lewis group carbohydrate antigens during Xenopus development.

We have examined the pattern of expression of the Lewis group carbohydrate antigens during the development of African toad Xenopus laevis. One of these antigens, Lewis x (Le(x), also known as SSEA-1), was previously shown to be involved in cell-cell adhesion in early mouse embryos and teratocarcinoma stem cells. Recently another member of these antigens, sialyl-Le(x), was found to be one of the major ligands for the selectin family of cell-cell adhesion molecules. In order to study the role of carbohydrate-mediated cell adhesion during Xenopus development, we first studied the expression pattern of the Le(x). We found that Le(x)was not expressed in early embryos, started to be expressed at the tail bud stage in anterior regions of the body such as the cement gland or head skin, and was gradually showed more posterial expression at later stages. At tadpole stage, it was also expressed on specific cell bodies in brain, and in axon region in brain and neural retina. Antibodies against Le(x)blocked neurite outgrowth in the explant culture of tadpole brain. One of the candidates for Le(x)carrier protein in the tadpole brain is a 200 kDa glycoprotein detected by Western blotting. In adult tissues, it was expressed in brain, testis, and gut, but not in kidney, lung, spleen, ovary, or muscle. We also examined the expression patterns of other Lewis group antigens. Among them, sialyl-Le(x)was expressed on endothelial cells and on leukocytes, suggesting the possibility that it functions as a ligand for selectin in Xenopus.

Bix4 is activated directly by VegT and mediates endoderm formation in Xenopus development.

The maternal T-box gene VegT, whose transcripts are restricted to the vegetal hemisphere of the Xenopus embryo, plays an essential role in early development. Depletion of maternal VegT transcripts causes embryos to develop with no endoderm, while vegetal blastomeres lose the ability to induce mesoderm (Zhang, J., Houston, D. W., King, M. L., Payne, C., Wylie, C. and Heasman, J. (1998) Cell 94, 515-524). The targets of VegT, a transcription activator, must therefore include genes involved both in the specification of endoderm and in the production of mesoderm-inducing signals. We recently reported that the upstream regulatory region of the homeobox-containing gene Bix4 contains T-box binding sites. Here we show that expression of Bix4 requires maternal VegT and that two T-box binding sites are necessary and sufficient for mesodermal and endodermal expression of reporter genes driven by the Bix4 promoter in transgenic Xenopus embryos. Remarkably, a single T-box binding site is able to act as a mesoderm-specific enhancer when placed upstream of a minimal promoter. Finally, we show that Bix4 rescues the formation of endodermal markers in embryos in which VegT transcripts have been ablated but does not restore the ability of vegetal pole blastomeres to induce mesoderm. These results demonstrate that Bix4 acts directly downstream of VegT to specify endodermal differentiation in Xenopus embryos.