LED-display for an intraocular microoptic system.

In this paper the design and fabrication of an LED array usable for an intraocular vision aid (IOVA) is presented. IOVA is intended for people with vastly opaque cornea caused by explosion or chemical burns. The LED array consists of 1024 single GaP/GaAsP LEDs. In a first step an 8 x 8 pixel LED array has been fabricated. This LED array is bonded to a CMOS driver circuit performing a miniaturized display by using flip-chip-technique. Thus each LED pixel can be driven separately and the generated light passes the substrate.

A CMOS clock and data recovery circuit for intraocular microsystems.

This paper presents the implementation of a clock and data recovery circuit (CDR) for intraocular microsystems. The CDR was designed to minimize chip area and power consumption and to recover the clock and data signals from the incoming data stream. Since the CDR has been designed without any external components it is well suited for being integrated in an intraocular microsystem. Simulation results show that this CDR works with power dissipation of less than 2.4 mW with a single 3.3 V power supply. The simulations are based on a 0.6 micron n-well CMOS single-polysilicon, three-metal technology.

Direct determination of particle-particle interactions in a 2D plasma dust crystal.

Particle interactions were directly determined from the lateral compression of two-dimensional plasma dust crystals confined in a parabolic potential well. Measured particle separations combined with an equation of state for the crystal were used to derive values for the particle screening length and the charge on the particles. The shape of the parabolic potential well was confirmed by observing trajectories of single particles falling within the well. No evidence of a lateral particle-particle binding was observed.

Limits to the development of fast neuromuscular transmission in zebrafish.

Zebrafish embryos have small and slow miniature end-plate currents (mEPCs), whereas only a few days later larval mEPCs are an order of magnitude larger and faster, being among the fastest of all neuromuscular synapses. To identify the bases for these changes we compared, in embryos and larvae, the properties and distributions of acetylcholine (ACh) receptors (AChRs) and acetylcholinesterase (AChE) as well as the ultrastructure of the developing neuromuscular junctions (NMJs). To mimic synaptic release, patches of muscle membrane were exposed briefly (for 1 ms) to a saturating concentration (10 mM) of ACh. The AChR deactivation kinetics were twice as slow in embryos compared with larvae. In both embryos and larvae, AChRs demonstrated open channel block by millimolar ACh, and this was detected during mEPCs, indicating that a high concentration of ACh is released at immature and mature NMJs. AChR and AChE distributions were compared using the selective fluorescently conjugated labels alpha-bungarotoxin and fasciculin 2, respectively. In larvae, punctate AChR clusters were detected whereas junctional AChE staining was less intense than that found at adult NMJs. Transmission electron microscopy revealed immature nerve endings in embryos that were closely juxtaposed to the surrounding muscle cells, whereas mature larval NMJs had a wider synaptic cleft with a conspicuous basal lamina over a limited region of synaptic contact. Our results indicate that ACh is released at high concentrations at immature NMJs, but its clearance is prolonged and the AChRs are dispersed, resulting in a slow mEPC time course until a mature cleft appears with densely packed faster AChRs and abundant AChE.

From efficacy to effectiveness: managing organisational change to improve health services for young people with deliberate self harm behaviour.

Repeat Deliberate Self Harm is a recognised risk factor for completed suicide and therefore reduction by effective health service response represents a valid contribution to suicide prevention. However, only a small fraction of people with deliberate self harm presentations to general health settings actually reach specialist mental health follow-up appointments. Therefore, even if responses at that point are known to be effective they do not make a significant contribution to reducing repeat self-harm overall. We describe health system organisational change strategies to improve health service engagement for the target group, and present data demonstrating the effectiveness of these strategies.

Synaptic drive to motoneurons during fictive swimming in the developing zebrafish.

The development of swimming behavior and the correlated activity patterns recorded in motoneurons during fictive swimming in paralyzed zebrafish larvae were examined and compared. Larvae were studied from when they hatch (after 2 days) and are first capable of locomotion to when they are active swimmers capable of capturing prey (after 4 days). High-speed (500 Hz) video imaging was used to make a basic behavioral characterization of swimming. At hatching and up to day 3, the larvae swam infrequently and in an undirected fashion. They displayed sustained bursts of contractions ('burst swimming') at an average frequency of 60-70 Hz that lasted from several seconds to a minute in duration. By day 4 the swimming had matured to a more frequent and less erratic "beat-and-glide" mode, with slower (approximately 35 Hz) beats of contractions for approximately 200 ms alternating with glides that were twice as long, lasting from just a few cycles to several minutes overall. In whole cell current-clamp recordings, motoneurons displayed similar excitatory synaptic activity and firing patterns, corresponding to either fictive burst swimming (day 2-3) or beat-and-glide swimming (day 4). The resting potentials were similar at all stages (about -70 mV) and the motoneurons were depolarized (to about -40 mV) with generally non-overshooting action potentials during fictive swimming. The frequency of sustained inputs during fictive burst swimming and of repetitive inputs during fictive beat-and glide swimming corresponded to the behavioral contraction patterns. Fictive swimming activity patterns were eliminated by application of glutamate antagonists (kynurenic acid or 6-cyano-7-nitroquinoxalene-2,3-dione and DL-2-amino-5-phosphonovaleric acid) and were modified but maintained in the presence of the glycinergic antagonist strychnine. The corresponding synaptic currents underlying the synaptic drive to motoneurons during fictive swimming could be isolated under voltage clamp and consisted of cationic [glutamatergic postsynaptic currents (PSCs)] and anionic inputs (glycinergic PSCs). Either sustained or interrupted patterns of PSCs were observed during fictive burst or beat-and-glide swimming, respectively. During beat-and-glide swimming, a tonic inward current and rhythmic glutamatergic PSCs (approximately 35 Hz) were observed. In contrast, bursts of glycinergic PSCs occurred at a higher frequency, resulting in a more tonic pattern with little evidence for synchronized activity. We conclude that a rhythmic glutamatergic synaptic drive underlies swimming and that a tonic, shunting glycinergic input acts to more closely match the membrane time constant to the fast synaptic drive.

Physiological properties of zebrafish embryonic red and white muscle fibers during early development.

The zebrafish is a model organism for studies of vertebrate muscle differentiation and development. However, an understanding of fish muscle physiology during this period is limited. We examined the membrane, contractile, electrical coupling, and synaptic properties of embryonic red (ER) and white (EW) muscle fibers in developing zebrafish from 1 to 5 days postfertilization. Resting membrane potentials were -73 mV in 1 day ER and -78 mV in 1 day EW muscle and depolarized 17 and 7 mV, respectively, by 5 days. Neither fiber type exhibited action potentials. Current-voltage relationships were linear in EW fibers and day 1 ER fibers but were outwardly rectifying in some ER fibers at 3 to 5 days. Both ER and EW fibers were contractile at all ages examined (1 to 5 days) and could follow trains of electrical stimulation of up to 30 Hz without fatiguing for up to 5 min. Synaptic activity consisting of miniature endplate potentials (mEPPs) was observed at the earliest ages examined (1.2-1. 4 days) in both ER and EW fibers. Synaptic activity increased in frequency, and mEPP amplitudes were larger by 5 days. Miniature EPP rise times and half-widths decreased in ER fibers by 5 days, while EW fiber mEPPs showed fast kinetics as early as 1.2-1.4 days. ER and EW muscle fibers showed extensive dye coupling but not heterologous (red-white) coupling. Dye coupling decreased by 3 days yet remained at 5 days. Somites were electrically coupling, and this allowed filtered synaptic potentials to spread from myotome to myotome. It is concluded that at early developmental stages the physiological properties of ER and EW muscle are similar but not identical and are optimized to the patterns of swimming observed at these stages.

Properties of miniature glutamatergic EPSCs in neurons of the locomotor regions of the developing zebrafish.

As a first step in understanding the development of synaptic activation in the locomotor network of the zebrafish, we examined the properties of spontaneous, glutamatergic miniature excitatory postsynaptic currents (mEPSCs). Whole cell patch-clamp recordings were obtained from visually identified hindbrain reticulospinal neurons and spinal motoneurons of curarized zebrafish 1-5 days postfertilization (larvae hatch after the 2nd day of embryogenesis). In the presence of tetrodotoxin (TTX) and blockers of inhibitory receptors (strychnine and picrotoxin), we detected fast glutamatergic mEPSCs that were blocked by the AMPA/kainate receptor-selective antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). At positive voltages or in the absence of Mg(2+), a second, slower component of the mEPSCs was revealed that the N-methyl-D-aspartate (NMDA) receptor-selective antagonist DL-2-amino-5-phosphonovalerate (AP-5) abolished. In the presence of both CNQX and AP-5, all mEPSCs were eliminated. The NMDA component of reticulospinal mEPSCs had a large single-channel conductance estimated to be 48 pS. Larval AMPA/kainate and NMDA components of the mEPSCs decayed with biexponential time courses that changed little during development. At all stages examined, approximately one-half of synapses had only NMDA responses (lacking AMPA/kainate receptors), whereas the remainder of the synapses were composed of a mixture of AMPA/kainate and NMDA receptors. There was an overall increase in the frequency and amplitude of mEPSCs with an NMDA component in reticulospinal (but not motoneurons) during development. These results indicate that glutamate is a prominent excitatory transmitter in the locomotor regions of the developing zebrafish and that it activates either NMDA receptors alone at functionally silent synapses or together with AMPA/kainate receptors.

In vivo recording from identifiable neurons of the locomotor network in the developing zebrafish.

The zebrafish is a popular model for developmental studies due to its accessibility by cellular, molecular and genetic approaches. As a complement to these other methods, we have devised an exposed hindbrain/spinal cord preparation in the curarized zebrafish embryo and larva that permits intracellular labeling and patch clamp recording from individually identified sensory neurons, motoneurons and interneurons in vivo. Regular bursts of synaptic potentials and action potentials were observed under whole-cell current clamp in embryonic motoneurons and in some identified interneurons. Larval neurons showed prolonged depolarizations with synaptically driven bursts of action potentials. Frequent spontaneous synaptic potentials were observed and synaptic currents were effectively space clamped. It is thus feasible to study in vivo the properties of identifiable neurons of the developing locomotor network in the zebrafish, including their synaptic activity, firing patterns and interconnections.

Excitability changes in sacral afferents innervating the urethra, perineum and hindlimb skin of the cat during micturition.

1. Excitability changes in afferents innervating the urethra, perineum and hindlimb were measured in decerebrated cats during micturition and in response to stimulation of lumbosacral afferents. Increases in excitability were interpreted as primary afferent depolarization (PAD) and decreases as primary afferent hyperpolarization. 2. Excitability increases were observed in 11 of 19 urethral pudendal afferents during micturition. Four of these 11 afferents showed an excitability increase during voiding. Seven of these showed a biphasic change with a decrease in excitability when sphincter activity resumed at the end of the void. Three of 19 afferents showed an excitability decrease during micturition and no change was detected in five afferents. 3. During micturition, the peak amplitude of urethral afferent-evoked excitatory postsynaptic potentials in seven of eight sphincter motoneurones was diminished to a mean of 36% of control values. 4. Eighty per cent of hindlimb cutaneous afferents and 50% of dorsal penile/clitoral and superficial perineal nerve afferents in the sacral cord showed increased excitability during voiding. No excitability increases were measured in 13 hindlimb cutaneous fibres examined in the lumbar segments. 5. PAD was observed in sacral urethral, perineal and hindlimb cutaneous afferents in response to electrical stimulation of other perineal, urethral, hindlimb cutaneous and group II muscle afferents. 6. It is concluded that control of transmission from urethral afferents by the micturition circuitry is different to that by sensory transmission from hindlimb and perineal regions during micturition. We hypothesize that more than one population of sacral PAD-mediating interneurones is involved.

Urethral pudendal afferent-evoked bladder and sphincter reflexes in decerebrate and acute spinal cats.

Electrical stimulation of the urethral sensory pudendal nerve in decerebrate or acute spinal cats was used to evoke micturition reflexes in animals that failed to respond to bladder distension. In the decerebrate animals, stimulation of urethral afferents evoked voiding characterized by a large bladder pressure increase coordinated with a simultaneous decrease in external urethral sphincter activity. In animals in which the spinal cord was transected between T10 and L6, electrical stimulation of the urethral afferents evoked small increases in bladder pressure that were insufficient to expel fluid but the contractions were coordinated with a decrease in external urethral sphincter activity. It was concluded that in addition to interacting with spinobulbospinal micturition pathways, urethral pudendal afferents may have direct access to a spinal circuitry that can coordinate bladder and sphincter activity.

Evidence for a strychnine-sensitive mechanism and glycine receptors involved in the control of urethral sphincter activity during micturition in the cat.

Micturition in the decerebrate cat is characterized by a coordinated bladder contraction and a simultaneous decrease in external urethral sphincter (EUS) efferent activity. Without the suppression of EUS activity, voiding is significantly impaired, resulting in a state sometimes referred to as bladder-sphincter dyssynergia. The aim of the present study was to determine whether glycinergic inhibition contributes to the suppression of EUS activity during micturition evoked by bladder distension or electrical stimulation of the pontine micturition center (PMC) in decerebrate cats. Using subconvulsive intravenous doses of strychnine (0.1-0.24 mg/kg), we examined changes in bladder and EUS electroneurographic (ENG) activity during micturition. Following subconvulsive doses of strychnine, tonic EUS ENG activity increased during bladder filling in five of six animals. In the presence of strychnine, it was possible to evoke reflex bladder contractions of similar duration and peak pressure to those observed before strychnine administration. However, there was an absence of suppression of EUS ENG activity during the bladder contractions in all the animals. To determine whether the changes in sphincter activity could be due to strychnine acting at glycine receptors on EUS motoneurons, sacral spinal tissue was processed for a structural protein (gephyrin) associated with the glycine receptor. Motoneurons in Onufs nucleus in S1 were identified using choline acetyltransferase immunohistochemistry and subsequently processed with a gephyrin monoclonal antibody. Abundant gephyrin labeling was evident throughout Onufs nucleus. Since Onufs nucleus is made up of EUS and other motoneuron populations, a sample of antidromically identified urethral and anal sphincter motoneurons were intracellularly labeled with tetramethylrhodamine dextran (TMR-D) and then processed with the gephyrin antibody. Using dual-beam confocal microscopy, gephyrin immunoreactivity was observed on the soma and proximal processes of individual EUS motoneurons in both male and female animals. It was concluded that a strychnine-sensitive mechanism contributes to the suppression of sphincter activity normally observed during voiding. Although glycinergic inhibition may affect several components of the circuitry responsible for micturition, it appears that the suppression of EUS motoneurons during micturition may be partly due to a direct glycinergic inhibition of the EUS motoneurons.

Do parenteral nutrition solutions with high concentrations of branched-chain amino acids offer significant benefits to stressed patients?

The critically ill, stressed patient has been characterized as having altered cellular metabolism. Altered protein metabolism is manifested as negative nitrogen balance, reduced whole-body protein synthesis, and increased proteolysis. An increased oxidation of the branched-chain amino acids (BCAA) leucine, isoleucine, and valine has also been observed. Exogenous administration of BCAA as part of a total parenteral nutrition (TPN) regimen has been proposed to compensate for the altered protein metabolism in the stressed patient by sparing endogenous sources of BCAA, thereby reducing skeletal muscle catabolism and increasing protein synthesis. Numerous clinical studies have been performed investigating this theory. The results are controversial. Differences in study outcomes appear to be related to study design, especially patient selection. Our review of those studies which were randomized, prospective, and controlled indicates that an improvement in nitrogen retention and visceral protein status can be achieved in stress-stratified patients who receive a TPN regimen containing a BCAA-enriched formula. The significance of these outcomes on morbidity, length of hospital stay, and mortality has not been evaluated.

The Autonomic Nervous System Response Inventory (ANSRI): discrimination among migraine headache sufferers, muscle contraction headache sufferers, and normal controls.

Twenty-three migraine headache sufferers, sixteen muscle contraction headache sufferers, and thirteen no headache control subjects were selected to test the discriminant validity of the ANSRI. Significant Chi-Squares indicated reliable differences among the groups in ANSRI F scale scores under the Anger condition and in P scale scores based on the means of items across four emotions (All E). Discriminant analyses yielded 69% correct classifications for All E P scales and 58% correct classifications for Anger F scales. All E P scale analyses revealed that the Muscle Tension scale separated the headache groups from the control group. A second function separated the three groups from each other, with the Cardiac, Respiration, and Gastrointestinal scales most responsible. Anger F scale analysis showed the Peripheral Vasoconstriction, Cardiac, and Pattern 2 scales separating the headache groups from each other and from the control group. Results demonstrated discriminant validity for the ANSRI, and were consistent with muscle tension and vasoconstriction as variables in muscle contraction and migraine headache, respectively.

Emergency operation in patients with symptomatic abdominal aortic aneurysms.

To assess the need for routine preoperative computerized tomography scanning to discern patients with rupture among those presenting with acutely symptomatic abdominal aortic aneurysms, a retrospective review was performed. During a 5-year period, all patients presenting with symptomatic aneurysm underwent emergency operation without preoperative computerized tomography. The mortality rate was not significantly different among patients with symptomatic, intact aneurysms undergoing emergency operation (3 percent) and those without symptoms having elective operation (5 percent). The mortality rate of patients with ruptured aneurysms was 68 percent. We concluded that the addition of preoperative computerized tomography to the clinical evaluation would not have improved these results. Furthermore, since it is expensive and delays emergency operation in patients with ruptured aneurysms, computerized tomography seems rarely indicated in symptomatic patients with obvious aneurysms.

Iron balances in infant nutrition.

Iron balance studies were performed in 17 full term male infants from their 3rd until their 17th week of life. The balance studies were made in the infant's home and comprised 5 periods with an interval of 3-4 weeks, each consisting of three 24-hour collections. Ten infants were breast-fed, 3 received an adapted infant formula (P1, iron content 1.1 mg/l) and 4 were given the same formula enriched with iron, copper and zinc (P2, iron content 10.35 mg/l). From the 3rd to the 17th week of life the breast-fed infants got a mean iron intake of 0.2 mg/kg body weight X 3 days and they retained 0.09 mg/kg b.w. X 3 days. The P1 group received 0.48 and 0.47 mg/kg b.w. X 3 days and retained -0.01 and -0.5 mg iron/kg b.w. X 3 days, while the P2 group had an intake from 5.04 to 6.38 mg b.w. X 3 days and retained between 1.13 and 3.66 mg iron/kg b.w. X 3 days. Comparing the 3 groups it can be concluded that the P1 group retained definitely less iron than the breast-fed group, whereas the P2 group retained 12 to 40 times more iron than the breast-fed babies.

Assessing the impact of psychological intervention on family practice clinic visits.

The effects of brief psychological intervention on rates of medical utilization were investigated in a family practice clinic. Patients referred to the psychology service served as experimental subjects. These subjects underwent psychological assessment and treatment from supervised clinical psychology externs in joint working arrangements with family practice residents. Control subjects were other family practice patients, matching the experimental group in age, sex, and race. The number of medical visits to the family practice clinic, specialty clinics, and the emergency room were the dependent measures. The study period was two years. Results indicated that the experimental subjects were more likely than control subjects to decrease medical utilization of the family practice clinic. The findings are consistent with earlier work showing an association between brief psychosocial intervention and decreases in the frequency of clinic visits. However, this study suggests that preinternship-level clinical psychology trainees can be equally as effective in providing this intervention as mental health professionals who have completed their training.

A clinical model for enhancing adolescent ego identity.

A model of identity formation based on the cognitive developmental stages of social perspective-taking is described. The model assumes that identity can be achieved through cognitive strategies of considering the self in relation to one friend, one's family, the peer group, and society. Two studies were undertaken to test the model. In Study 1, 28 college students participated in a two week program in which the experimental students were asked to take the perspective of each social entity above and then to consider how the self was like and unlike these. The experimentals gained more than the controls on the identity subscale of Rasmussen's EIS. Fifty-nine college students participated in Study 2 which used a more sensitive discrepancy score measure on a revised identity scale. Discrepancy scores in an analysis of covariance indicated that the experimentals were higher than the controls at post-test and at a one month follow-up.

Mixed meal tolerance test and reactive hypoglycemia.

Twenty-six patients with symptoms suggestive of postprandial hypoglycemia were investigated by oral glucose tolerance test (OGTT). During the OGTT, symptomatic hypoglycemia occurred in 10 (38.5%). Nine of these 10 sugjects were given mixed meal tolerance tests (MMTT) and symptomatic hypoglycemia failed to occur in any case. During the OGTT the nadir glucose was significantly lower than that during MMTT (44.1 +/- 1.5 vs. 77.3 +/- 4.8 mg/dl +/- SEM, respectively; p less than 0.0005). Serum insulin during MMTT peaked significantly earlier than during OGTT (46.7 +/- 7.3 vs. 86.7 +/- 11.7 minutes (SEM, respectively; p less than 0.0125). The early secretion of insulin during MMTT may explain the lack of symptomatic hypoglycemia in these patients. We conclude that reactive hypoglycemia, when tested by a more natural stimulus (such as mixed meal) rather than by OGTT, is uncommon.