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To enhance the understanding of differentiation patterns and bone formation capacity of hESCs, we determined (1) the temporal pattern of osteoblastic differentiation of human embryonic stem cell derived mesenchymal stem cells (hESC-MSCs), (2) the influence of a three-dimensional matrix on the osteogenic differentiation of hESC-MSCs in long-term culture, and (3) the bone-forming capacity of osteoblast-like cells derived from hESC-MSCs in calvarial defects. Incubation of hESC-MSCs in osteogenic medium induced osteoblastic differentiation of hESC-MSCs into mature osteoblasts in a similar chronological pattern to human bone marrow stromal cells and primary osteoblasts. Osteogenic differentiation was enhanced by culturing the cells on three-dimensional collagen scaffolds. Fluorescent-activated cell sorting of alkaline phosphatase expressing...
The [14C]-2-deoxyglucose (2-DG) autoradiographic technique was used to study how auditory-related metabolic activity changes with deafness, and how chronic electrical stimulation of the deafened system may modify these changes. Guinea pigs were deafened by administration of kanamycin and ethacrynic acid. After nine weeks of deafness, the basal unstimulated uptake of 2-DG in the inferior colliculus (IC) was lower than in normal hearing control animals. 100 [mu]A of acute cochlear electrical stimulation significantly increased 2-DG uptake in normal hearing animals but did not evoke a significant increase in four or nine week deafened animals. Electrically elicited 2-DG uptake in the IC is therefore depressed by prolonged deafness. In a second series of experiments, after four weeks of deafness, animals were chronically electrically stimu...
Detection thresholds for biphasic symmetric pulses were measured in fourteen human subjects implanted with the Cochlear Corporation Nucleus 22 Implant. The effects of phase duration on thresholds were studied using single pulses, and 500 ms pulse trains at 100 pps. Psychophysical detection thresholds decreased as a function of phase duration with a change in slope at approximately 0.5 ms/phase. Mean single-pulse and pulse-train slopes were -3.60 and -4.25 dB/doubling of phase duration for pulse durations of less than about 0.5 ms/phase. For pulse durations greater than 0.5 ms/phase, mean slopes were -5.71 and -7.54 dB/doubling for single pulses and pulse trains, respectively. Thresholds for pulse trains decreased as a function of stimulus duration for durations up to at least 300 ms, with the rate of decrease being dependent on the pha...
This study evaluated the psychosomatogenic family model as an etiological theory of psychosomatic symptomatology among chronically ill asthmatic children. The model postulates that four family transactional characteristics--enmeshment, overprotectiveness, lack of conflict resolution, and rigidity--nurture psychosomatic symptoms among chronically ill children. These four characteristics were measured in the families of 42 asthmatic children, and the severity of each child's asthmatic condition was assessed through parents' ratings, medical records, and physicians' clinical judgements. Factor analysis was used to create composite variables of severity, and multiple regression analysis was used to assess the relationships between the four family characteristics and six dimensions of severity of the asthmatic condition. No evidence was fou...
The perilymphatic space of the guinea pig cochlea was perfused with various concentrations of Ca2+, Mg2+ and gentamicin. Cochlear microphonic potentials (CM) were essentially stable when solutions contained 1 mM Ca2+ plus 2 mM Mg2+, 1 mM Ca2+ and no Mg2+ or when both Ca2+ and Mg2+ were omitted. In the absence of Ca2+, the presence of 2 mM Mg2+ or 1 mM EGTA markedly decreased CM.Addition of 3 mM gentamicin lowered CM, the magnitude of the effect depending on the concentration of Ca2+ present. Ten mM Ca2+ eliminated the action of the drug. In the absence of Ca2+, the effects of Mg2+ and gentamicin were additive.When the application of gentamicin was followed by perfusion with drug-free medium, suppression of CM by 3 mM drug could be reversed with 10 mM Ca2+ but not with 1 mM Ca2+. Loss of CM was irreversible when administration of 10 mM ...
Impairment of the cochlear blood supply of guinea pigs was induced in order to study the effects of hypoxia on the cochlear action potentials. The oxygenation of the cochlear structures was decreased by perfusing the ears with polycythemic hyperviscous blood. The validity of using this model of cochlear blood flow was based on the propensity of the blood to flow in a laminar way. Because of the streamlined flow pattern, the blood supplied by the two vertebral arteries does not mix within the common trunk of the basilar artery. The retrograde injection of polycythemic hyperviscous blood into one vertebral artery will affect the ear on the injected side only. The high viscosity of the polycythemic blood decreases the rate of flow of blood through the cochlear vessels; the high oxygen content of this blood, however, avoids hypoxia of the ...
A method for the continuous perfusion of the perilymphatic space of the inner ear in the guinea pig is described. Artificial perilymph is supplied to the cochlea and drained away through a tubing system while flow rates from 10 [mu]l/min to 0.3 ml/min are established by gravity syphon pressure. Techniques are also presented which allow control over the temperature of the perfusate and over the level of dissolved oxygen in the perfusate. Along with these variables, the pH of the artificial perilymph can be manipulated and various drugs can be added to the perfusate to test their effect on the inner ear. The function of the inner ear is monitored by continuous recording of the sound evoked bioelectric potentials, the cochlear microphonic and the compound action potential. The cochlear perfusion technique has many applications in the stud...
Picogram quantities of norepinephrine were found in cochlear regions of pigmented and non-pigmented rats. These regions of the cochlea were the modiolus, organ of Corti-osseous spiral lamina and the lateral cochlear wall. The content of norepinephrine in the modiolus and lateral cochlear wall of the pigmented rat was significantly greater than that in areas of the non-pigmented rat. In contrast, there was no statistical difference between the norepinephrine content of the organ of Corti-osseous spiral lamina region of the pigmented rat and that of the albino rat. Since a major difference between the pigmented and albino rats is the presence of melanin-containing melanocytes in the modiolus and lateral cochlear wall region of the pigmented animals, it is possible that norepinephrine is stored in cochlear melanocytes.
The effect of loud sound on the perilymphatic oxygen tension was studied in anesthetized guinea pigs. Pure tone (4 kHz) and broad-band noise were given at 85-130 dB SPL for 3-8 min. No effects were seen either in the animals exposed to pure tone or in the animals exposed to 85 dB broad-band noise. In the animals exposed to noise at 130 dB SPL both increases and decreases of perilymphatic oxygen were measured but the changes were only of about 12% or less. The response to anoxia was normal. In animals with hypotension (PO2 fluctuated with the blood pressure. When the sound was delivered directly into the opened bulla the measured PO2 dropped immediately but was found to be caused by the cooling effect of an air current produced by the noise. Flushing the opened bulla with nitrogen, air or oxygen caused the same temperature-induced drop ...
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