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Frequency dependence of synchronization of cochlear nerve fibers in the alligator lizard: evidence for a cochlear origin of timing and non-timing neural pathways.

The dependence of synchronization of spike discharges on tone frequency was measured in cochlear nerve fibers of anesthetized alligator lizards at 21 degrees C. Synchronization measures were based on the fundamental component of a Fourier analysis of the instantaneous discharge rate in response to tone bursts. Measurements were obtained from fibers innervating hair cells in both the region of the cochlea that contains a tectorial membrane (tectorial fibers) and the region where hair-cell stereocilia are free-standing in scala media (free-standing fibers). Both rate and synchronization tuning-curves were measured automatically as a function of tone frequency. For tectorial fibers, the shapes of synchronization tuning-curves are roughly similar to the shapes of rate tuning-curves: the characteristic frequencies (CF's) of both curves are approximately equal. For free-standing fibers, the shape of synchronization tuning-curves differ markedly from those of rate tuning-curves. The CF's of synchronization and rate tuning-curves differ - the ranges are 0.2-0.6 kHz and 1-4 kHz, respectively - and the two CF's are uncorrelated. Synchronization filter-functions, which are contours of synchronization index at constant average discharge rate, were measured as a function of tone frequency for both tectorial and free-standing fibers. These synchronization filter-functions have the shapes of lowpass filters. For the populations of tectorial fibers and of free-standing fibers taken separately, these functions are independent of CF. The corner frequency of these functions is 0.50 +/- 0.038 kHz for tectorial fibers and 0.37 +/- 0.037 kHz for free-standing fibers. We conclude that these populations are characterized by different synchronization filters. For free-standing fibers, synchronization filter-functions measured at average driven discharge rates of about 20 and 40 spikes/s do not differ appreciably, and the high-frequency slope is -80 to -115 dB/decade. The results show that tectorial fibers encode timing information for low-level stimuli, whereas free-standing fibers do not. It is proposed that in the alligator lizard, neural pathways that encode timing information originate in the tectorial region and those that encode non-timing information originate in the free-standing region.

Acoustic Stimulation↗

Fiber counting and analysis in the diagnosis of asbestos-related disease.

Analysis of numbers and types of asbestos fibers present in lung tissue may provide insights into the pathogenesis of asbestos-induced disease, as well as diagnostic information concerning the relationship of a given lesion to asbestos exposure. This type of analysis requires extraction of fibers and asbestos bodies from lung tissue, preferably by means of a digestion-and-concentration technique, and examination with a combination of electron optical techniques, including electron diffraction and energy-dispersive x-ray spectroscopy. The combination permits definitive identification of asbestos fibers. Asbestos bodies have been shown to contain asbestos no matter what population they are found in, but they appear to be of value in ascertaining unusual exposure only when present in very large numbers. Numbers of asbestos bodies markedly underestimate total numbers of fibers present in lung. In patients from the general population, the mean number of asbestos fibers is about 1 X 10(6)/g dry lung; of this number, more than 80 per cent are fibers of chrysotile less than 5 microns long. Patients in the general population who have pleural plaques have about the same total number of fibers, but their lungs contain about a 50-fold increase in long thin amphibole fibers of commercial origin. Patients who have asbestosis and most patients who have mesothelioma have 100 to 200 X 10(6) fibers/g dry lung; the grade of asbestosis appears to be related to total fiber content. Occasional patients may develop mesotheliomas with much smaller fiber burdens. Both benign and malignant pleural diseases appear to be closely related to the presence of long thin amphibole fibers. Analysis of pulmonary fiber burden suggest that asbestos-related disease is not merely a matter of total numbers of fibers present, but that factors such as fiber type and size are equally important.

Asbestos↗

1H-nuclear magnetic resonance evidence for acto-myosin-dependent structural changes of the intracellular water of frog skeletal muscle fiber.

The proton-spin relaxation process of the intracellular water in intact-relaxed and skinned-rigor fibers of frog skeletal muscle was studied for slack and stretched fibers by use of 1H-nuclear magnetic resonance technique. The longitudinal and transverse proton-spin relaxation processes of the intracellular water of intact-relaxed and skinned-rigor fibers were composed of a single- and multi-exponential processes, respectively. The longitudinal relaxation process was almost the same in slack as well as in stretched fibers for both intact-relaxed and skinned-rigor fibers. On the other hand, the transverse relaxation process was slightly but significantly faster in stretched than in slack fibers in the case of skinned-rigor fibers while it was almost the same in slack and in stretched fibers in the case of intact-relaxed fibers. As the overlap between actin and myosin filaments is maximal in slack fibers and minimal in stretched fibers, these results indicate that the intracellular water located in the overlap region is less structured in rigor fibers than that in relaxed fibers. This suggests that the rigor crossbridge formation disrupts the structured water bound to myosin and actin filaments in muscle fiber.

Actins↗

Mammalian skeletal muscle fibers distinguished by contents of phosphocreatine, ATP, and Pi.

We tested the proposition that muscle cell types have different contents of phosphocreatine (PCr), ATP, and Pi by 31P NMR spectroscopy and HPLC analyses of adult rat and mouse muscles containing various volume fractions of different fiber types. There was a 2-fold difference in the PCr content between muscles with a high volume fraction of fiber types 1 and 2x versus those with fast-twitch (types 2a and 2b) fiber types. Pi content was low, and PCr and ATP contents were high in muscles with large contents of type 2b and 2a fibers; the reverse was true in muscles with a large volume fraction of type 1 and 2x fibers. There is a large range in the Pi/PCr ratios in normal resting muscles, from less than 0.05 in type 2 to 0.51 in type 1 fibers, depending upon the distribution of their component fiber types. In all muscles, the peak area resulting from the beta phosphate of ATP constituted approximately 13% of the sum of all peak areas observable in the 31P spectrum. Fiber types 2a and 2b were not distinguishable, and the content of type 2x fibers was similar to type 1 fibers. From the profile of these metabolites, we could distinguish only two classes of fibers. For type 2a and 2b fibers, the intracellular concentrations were 8 mM ATP, 39 mM total creatine, 32 mM PCr, 0.8 mM Pi, and 8 microM ADP. For type 1 and 2x fibers, these quantities were 5 mM ATP, 23 mM total creatine, 16 mM PCr, 6 mM Pi, and 11 microM ADP. Thus our results establish an additional criterion upon which to distinguish skeletal muscle cells, one based on the resting content of bioenergetically important metabolites. These results also provide the basis for estimating skeletal muscle fiber-type composition from noninvasive NMR spectroscopic data.

Adenosine Triphosphate↗

Microfibril-associated MAGP-2 stimulates elastic fiber assembly.

Elastic fibers are complex structures composed of a tropoelastin inner core and microfibril outer mantle guiding tropoelastin deposition. Microfibrillar proteins mainly include fibrillins and microfibril-associated glycoproteins (MAGPs). MAGP-2 exhibits developmental expression peaking at elastic fiber onset, suggesting that MAGP-2 mediates elastic fiber assembly. To determine whether MAGP-2 regulates elastic fiber assembly, we used an in vitro model featuring doxycycline-regulated cells conditionally overexpressing exogenous MAGP-2 and constitutively expressing enhanced green fluorescent protein-tagged tropoelastin. Analysis by immunofluorescent staining showed that MAGP-2 overexpression dramatically increased elastic fibers levels, independently of extracellular levels of soluble tropoelastin, indicating that MAGP-2 stimulates elastic fiber assembly. This was associated with increased levels of matrix-associated MAGP-2. Electron microscopy showed that MAGP-2 specifically associates with microfibrils and that elastin globules primarily colocalize with MAGP-2-associated microfibrils, suggesting that microfibril-associated MAGP-2 facilitates elastic fiber assembly. MAGP-2 overexpression did not change levels of matrix-associated fibrillin-1, MAGP-1, fibulin-2, fibulin-5, or emilin-1, suggesting that microfibrils and other elastic fiber-associated proteins known to regulate elastogenesis do not mediate MAGP-2-induced elastic fiber assembly. Moreover, mutation analysis showed that MAGP-2 does not stimulate elastic fiber assembly through its RGD motif, suggesting that integrin receptor binding does not mediate MAGP-2-induced elastic fiber assembly. Because MAGP-2 interacts with Jagged-1 that controls cell-matrix interaction and cell motility, two key factors in elastic fiber macroassembly, microfibril-associated MAGP-2 may stimulate elastic fiber macroassembly by targeting the release of elastin globules from the cell membrane onto developing elastic fibers.

Animals↗

Effect of fiber length on glass microfiber cytotoxicity.

Fiber length has been implicated as a determinant of fiber toxicity. Fibers of narrowly defined length can be generated by dielectrophoretic classifiers. Since the quantities of fibers produced are very small, we developed a rat alveolar macrophage microculture system to study the toxicity of these samples. The objective of this study was to examine the role of fiber length on the cytotoxicity of Manville code 100 (JM-100) fibers. Rat alveolar macrophages were cultured with 0-500 microg/ml of 5 lengths of JM-100 fibers on 96-well plates. After 18 h, well supernatants were removed and lactate dehydrogenase (LDH) activity was measured to assess cell damage. Chemiluminescence (CL), an assessment of macrophage function, was measured by adding lucigenin with or without zymosan, a particulate stimulus, to appropriate wells. For each fiber length the effects were concentration dependent: CL declined and LDH rose with increasing fiber concentration. Comparing the effects of different lengths showed the greatest toxicity from a relatively long fiber sample (mean length = 17 microm). Microscopic examination of the interaction of fibers with macrophages revealed multiple macrophages attached along the length of the long fibers. This suggests that frustrated, or incomplete, phagocytosis may be a factor in the increased toxicity of longer fibers. Overall the results demonstrate that length is an important determinant of toxicity for JM-100 fibers.

Acridines↗

A study of the effect of chrysotile fiber surface composition on genotoxicity in vitro.

Chrysotile fibers (NIEHS intermediate length) were treated with ultrapure HCl to alter the fiber surface chemistry without substantially changing fiber morphology or dimensions. The objective of the study was to determine whether fiber surface chemistry is an important variable in fiber genotoxicity in vitro. The modified fibers, along with native chrysotile fibers, were used to challenge Chinese hamster lung fibroblasts (V79) in vitro using the micronucleus induction genotoxicity assay. Fiber dimensions were assessed using scanning electron microscopy by measuring the distribution of fiber lengths in 3 length ranges: less than 3 microm, 3-10 microm, and greater than 10 microm. For both treated and native fiber samples, 500 fibers were examined. Results indicate that acid-treated fibers were about 20% shorter than untreated chrysotile. Surface chemistry alterations were verified by zeta-potential reversal, x-ray photoelectron spectroscopy (XPS), and scanning electron microscopy/energy-dispersive x-ray spectroscopy (SEM-EDS) elemental analysis. Scanning Auger spectrometry indicated the presence of Mg, O, and Si in both treated and native chrysotile samples, which confirmed the surface purity of both fiber samples. Both XPS and SEM-EDS analysis demonstrated substantial depletion of Mg from fiber surfaces. Results of the micronucleus assay showed a positive concentration-related response for both samples, with toxicity evident only at the highest concentration. No significant difference was found for the treated and untreated chrysotile samples. These results indicate that the surface chemistry is not an important variable in the in vitro genotoxicity of chrysotile asbestos in V79 cells as detected by the micronucleus assay under the conditions used in this study, and support a model of chemically nonspecific chromosomal and spindle damage effects.

Animals↗

Synthetic vitreous fibers: a review of toxicology research and its impact on hazard classification.

Because the inhalation of asbestos, a naturally occurring, inorganic fibrous material, is associated with lung fibrosis and thoracic cancers, concerns have been raised about the possible health effects of synthetic vitreous fibers (SVFs). SVFs include a very broad variety of inorganic fibrous materials with an amorphous molecular structure. Traditionally, SVFs have been divided into three subcategories based on composition: fiberglass, mineral wool (rock, stone, and slag wools), and refractory ceramic fiber. For more than 50 years, the toxicologic potential of SVFs has been researched extensively using human epidemiology and a variety of laboratory studies. Here we review the research and its impact on hazard classification and regulation of SVFs. Large, ongoing epidemiology studies of SVF manufacturing workers have provided very little evidence of harmful effects in humans. Several decades of research using rodents exposed by inhalation have confirmed that SVF pulmonary effects are determined by the "Three D's", fiber dose (lung), dimension, and durability. Lung dose over time is determined by fiber deposition and biopersistence in the lung. Deposition is inversely related to fiber diameter. Biopersistence is directly related to fiber length and inversely related to fiber dissolution and fragmentation rates. Inhaled short fibers are cleared from the lung relatively quickly by mobile phagocytic cells, but long fibers persist until they dissolve or fragment. In contrast to asbestos, most of the SVFs tested in rodent inhalation studies cleared rapidly from the lung (were nonbiopersistent) and were innocuous. However, several relativley biopersistent SVFs induced chronic inflammation, lung scarring (fibrosis), and thoracic neoplasms. Thus, biopersistence of fibers is now generally recognized as a key determinant of the toxicologic potential of SVFs. In vitro dissolution of fibers in simulated extracellular fluid correlates fairly well with fiber biopersistence in the lung and pulmonary toxicity, but several exceptions suggest that biopersistence involves more than dissolution rate. Research demonstrating the relationship between biopersistence and SVF toxicity has provided a scientific basis for hazard classification and regulation of SVFs. For a nonhazardous classification, legislation recently passed by the European Union requires a respirable insulation wool to have a low lung-biopersistence or be noncarcinogenic in laboratory rats. U.S. fiberglass and mineral wool industries and the Occupational Health and Safety Administration (OSHA) have formed a voluntary Health and Safety Partnership Program (HSPP) that include: a voluntary permissible exposure level (PEL) in the workplace of 1 fiber/cc, a respiratory protection program for specified tasks, continued workplace air monitoring, and, where possible, the development of fiber formulations that do not persist in the lung. RCF manufacturers have implemented a Product Stewardship Program that includes: a recommended exposure guideline of 0.5 fibers/cc; a 5-year workplace air monitoring program; and research into the development of high-temperature-resistant, biosoluble fibers.

Animals↗

Intrinsic optical and passive electrical properties of cut frog twitch fibers.

This article describes a new apparatus for making simultaneous optical measurements on single muscle fibers at three different wavelengths and two planes of linear polarization. There are two modes of operation: mode 1 measures the individual absorbances of light linearly polarized along and perpendicular to the fiber axis, and mode 2 measures retardation (or birefringence) and the average of the two absorbance components. Although some intact frog twitch fibers were studied, most experiments used cut fibers (Hille, B., and D. T. Campbell. 1976. Journal of General Physiology. 67:265-293) mounted in a double-Vaseline-gap chamber (Kovacs, L., E. Rios, and M. F. Schneider. 1983. Journal of Physiology. 343:161-196). The end-pool segments were usually exposed for 2 min to 0.01% saponin. This procedure, used in subsequent experiments to make the external membranes in the end pools permeable to Ca indicators (Maylie, J., M. Irving, N. L. Sizto, G. Boyarsky, and W. K. Chandler. 1987. Journal of General Physiology. 89:145-176; Maylie, J., M. Irving, N. L. Sizto, and W. K. Chandler. 1987. Journal of General Physiology. 89:41-143), was routinely employed so that all our cut fiber results would be comparable. A simple method, which does not require microelectrodes, allowed continual estimation of a fiber's membrane (rm) and internal longitudinal (ri) resistances as well as the external resistance (re) under the Vaseline seals. The values of rm and ri obtained from cut fibers with this method agree reasonably well with values obtained from intact fibers using microelectrode techniques. Optical measurements were made on resting and action potential-stimulated fibers. The intrinsic fiber absorbance, defined operationally as log10 of the ratio of incident light to transmitted light intensity, was similar in intact and cut preparations, as were the changes that accompanied stimulation. On the other hand, the resting birefringence and the peak of the active change in cut fibers were, respectively, only 0.8 and 0.7 times the corresponding values in intact fibers. Both the amplitude and the half-width of the active retardation signal increased considerably during the time course of cut fiber experiments; a twofold increase in 2 h was not unusual. Such changes are probably due to a progressive alteration in the internal state of the cut fibers.

Action Potentials↗

Relationships between alkali light-chain complement and myosin heavy-chain isoforms in single fast-twitch fibers of rat and rabbit.

The present study compares the alkali myosin light chain (LC) complement of the fast fiber types IIB, IID and IIA in single fibers from rat muscle, as well as in type IID and type IIA fibers from rabbit muscle. Single fibers were classified according to their electrophoretically determined myosin heavy chain (HC) isoforms, HCIIb, HCIId, and HCIIa. Alkali myosin light chains were analysed by densitometric evaluation of two-dimensional electrophoresis performed on extracts from the same fibers. On the average, the fraction of LC3f, i.e. LC3f/(LC1f+LC3f), was highest in type IIB fibers and lowest in type IIA fibers. Type IID fibers occupied an intermediate position. Also in the rabbit, type IID fibers displayed a higher fraction of LC3f than type IIA fibers. Large scattering of the LC3f fraction in IIB, IID, and IIA fibers indicated that each fiber type is composed of fibers identical with regard to their specific myosin heavy chain complement, but heterogeneous with regard to their fast alkali light chain composition and the resulting light-chain-based isomyosins. It is suggested that the variable proportions of the two alkali light chains in the three fast fiber populations serve as a fine tuning of contractile velocities within the ranges determined by the three fast myosin heavy-chain isoforms.

Animals↗

Early changes in fiber profile and capillary density in long-term stimulated muscles.

Predominantly fast skeletal muscles of rabbits [tibialis anterior (TA), extensor digitorum longus (EDL)] were stimulated at a frequency naturally occurring in nerves to slow muscles (10 Hz continuously) for 8 h/day for 2--4 days. Such stimulation is known to convert all glycolytic fibers to oxidative and to increase capillary density. Our aim was to study early stages of conversion to investigate the factors responsible for the changes. Staining of quick-frozen sections for myosin ATPase, succinic dehydrogenase, and alkaline phosphatase was used to study the distribution of different fiber types and to measure fiber cross-sectional areas, capillaries per square millimeter, and capillary-to-fiber ratios in each fiber category. TA but not EDL showed conversion of fast glycolytic to fast oxidative fibers after 2 days, more after 4 days of stimulation. In both muscles, the largest fast glycolytic fibers were diminished in number after stimulation. There was significant increase in total capillaries per square millimeter after 4 days and some increase after 2 days of stimulation. The increase in capillaries per square millimeter exceeded the increase in the number of fibers per square millimeter, and since there was no change in mean fiber area, the increase is attributed to capillary growth. In EDL, there was an increase in the number of capillaries supplying both fast glycolytic and fast oxidative fibers, suggesting that capillary growth precedes fiber type conversion. In TA, the number of capillaries supplying fast oxidative fibers was increased but that to fast glycolytic fibers, was not. This is consistent with capillary growth simultaneous with or following fiber conversion. In both TA and EDL the number of capillaries perfused after contraction was higher in stimulated muscles, suggesting that increased capillary flow contributed to capillary growth.

Adenosine Triphosphatases↗

Critical role of glass fiber length in TNF-alpha production and transcription factor activation in macrophages.

Recent studies have demonstrated that dielectrophoresis is an efficient method for the separation of fibers according to fiber length. This method allows the investigation of fiber-cell interactions with fiber samples of the same composition but of different lengths. In the present study, we analyzed the effects of length on the interaction between glass fibers and macrophages by focusing on production of the inflammatory cytokine tumor necrosis factor (TNF)-alpha in a mouse macrophage cell line (RAW 264.7). The underlying molecular mechanisms controlling TNF-alpha production were investigated at the gene transcription level. The results show that glass fibers induced TNF-alpha production in macrophages and that this induction was associated with activation of the gene promoter. Activation of the transcription factor nuclear factor (NF)-kappaB was responsible for this induced promoter activity. The inhibition of both TNF-alpha production and NF-kappaB activation by N-acetyl-L-cysteine, an antioxidant, indicates that generation of oxidants may contribute to the induction of this cytokine and activation of this transcription factor by glass fibers. Long fibers (17 micrometer) were significantly more potent than short fibers (7 micrometer) in inducing NF-kappaB activation, the gene promoter activity, and the production of TNF-alpha. This fiber length-dependent difference in the stimulatory potency correlated with the fact that macrophages were able to completely engulf short glass fibers, whereas phagocytosis of long glass fibers was incomplete. These results suggest that fiber length plays a critical role in the potential pathogenicity of glass fibers.

Acetylcysteine↗

Induction of long-term potentiation at hippocampal mossy-fiber synapses follows a Hebbian rule.

1. The induction of long-term potentiation (LTP) at hippocampal mossy-fiber synapses requires an increase in postsynaptic [Ca2+]i but is independent of N-methyl-D-aspartate (NMDA) receptor activation. Voltage-gated Ca2+ channels have been proposed as one alternative source for raising [Ca2+]i during the induction of LTP. We tested the hypothesis that voltage-gated Ca2+ channel activation could mediate the induction of LTP by examining whether 1) the induction of mossy-fiber LTP was dependent on postsynaptic depolarization and 2) depolarization alone, of a magnitude presumably capable of activating Ca2+ channels, was sufficient to induce LTP. 2. Intracellular recordings were made from rat CA3 pyramidal cells in the hippocampal slice preparation under both current- and voltage-clamp conditions. Mossy-fiber postsynaptic potentials and currents were recorded before and after high-frequency stimulation (HFS) in the presence of 20-50 microM D-2-amino-5-phosphonovaleric acid (D-APV), an NMDA-receptor antagonist. 3. Voltage clamping of CA3 neurons between -80 and -100 mV during HFS reversibly blocked the induction of mossy-fiber LTP. Conversely, HFS paired with depolarizing-current steps under current clamp increased the magnitude of LTP compared with controls. These results indicate that mossy-fiber LTP is dependent on postsynaptic depolarization, and presynaptic activation alone was not sufficient to induce mossy-fiber LTP. 4. Depolarizing-current injections, which presumably depolarized CA3 cells to potentials sufficient to activate voltage-gated Ca2+ channels, had no effect on mossy-fiber synaptic responses. These results suggest that synaptic activation, in addition to postsynaptic depolarization, is required for the induction of mossy-fiber LTP. 5. Single mossy-fiber afferent volleys were also paired with depolarizing-current pulses. In the presence of APV, pairing of single-mossy-fiber excitatory postsynaptic potentials (EPSPs) with postsynaptic depolarization did not potentiate synaptic responses, suggesting that some form of HFS is also required for mossy-fiber LTP. In the absence of APV, however, the contamination of mossy-fiber synaptic responses by CA3-recurrent inputs resulted in some potentiation. 6. These results suggest that the induction of mossy-fiber LTP is dependent on both pre- and postsynaptic activity and thus follows a Hebbian rule for synaptic modification. In contrast to that demonstrated at Schaffer-collateral-commissural synapses, however, the induction of mossy-fiber LTP may require HFS in addition to postsynaptic depolarization.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Organization and function of the collagen fiber system in the human temporomandibular joint disk and its attachments.

The collagen fiber organization in the anterior band (AB), intermediate zone (IZ) and posterior band (PB) of the disk of the human jaw joint disk and in its attachments was studied under the polarizing microscope. Observations were made on serially sectioned joints (n = 6) and three sets of disk samples sectioned along mutually perpendicular planes (n = 21). The collagen fiber bundles in all disk regions branch and join or are decussated with other bundles. The fibers of the IZ were oriented generally parallel to the disk surfaces. Most of these fibers extend into the AB and PB where they either join with transverse or vertically oriented fiber groups or pass through the bands into the disk attachments. The lateral branching/decussation angles of fibers in the IZ and adjacent regions of the AB and PB were measured in the central region of four disks. The mean angle for all regions was 28 +/- 13 degrees . The mean angles in the region transitional between the IZ and AB and in the anterior region of the IZ were significantly greater than those in the posterior part of the IZ (p = 0.001 and 0.050 for the two comparisons). The large caliber, vertically oriented fibers in the AB, IZ and PB were counted in nine specimens. Disk dimensions were also measured in these specimens. The vertically and transversely oriented bundles were more frequent in the band regions. The number of vertically oriented fiber bundles varied within and between the disk bands. Vertical fiber number in the AB was greater laterally than medially (p = 0.04). In the PB the fiber number was greater than in either the AB or IZ (p = 0.000 for both comparisons) and within the PB itself the fiber number was greater in its thicker, medial half (p = 0.014). The fiber number in the AB and IZ was not different. The thickness of the AB, IZ and PB and disk length was measured in sections located laterally, centrally and medially. No difference in disk length was found across these planes. Statistically significant differences were found in regional disk thickness. In all of the mediolateral planes, the AB and PB were thicker than the IZ and the PB was thicker than the AB (p = 0.000 for all comparisons). The IZ was thicker medially than laterally (p = 0.034). The PB was thicker centrally and medially than laterally (p = 0.002 and 0.001, respectively). PB thickness in its central and medial regions was not different. The findings, combined with other evidence, suggest that the fiber system of the disk serves a stress distribution function and that within the AB and PB, the regions containing the greatest number of vertical fibers may also be the sites of greatest compressive stress during jaw function.

Collagen↗

Beta-adrenergic receptor distribution among muscle fiber types and resistance arterioles of white, red, and intermediate skeletal muscle.

The pathophysiological effects of congestive heart failure and physiological effects of exercise training on skeletal muscle may be mediated in part by modulation of beta-adrenergic receptor density. To shed light on the physiological role of skeletal muscle beta-receptors, their density and distribution were characterized in muscle fibers and resistance arterioles of whole tissue slices of three rat hindquarter muscles differing markedly in fiber type composition and capacities for oxidative metabolism and vasodilatation. Binding isotherms and quantitative light microscopic autoradiographic localization of receptors were performed by incubating tissue slices in selected concentrations of [125I]cyanopindolol with and without 10(-5) M l-propranolol. Muscle fiber types were delineated in adjacent sections by histochemical staining of myofibrillar ATPase activity at pH 4.5-4.55. The total tissue content of receptors (Bmax) was nearly threefold greater in the soleus, a muscle consisting almost entirely of slow-twitch (type I) fibers than in superficial white vastus lateralis, a muscle composed of greater than 95% fast-twitch (type IIb) fibers. Bmax was intermediate in gastrocnemius, a mixed fiber muscle (all differences p less than 0.01). Receptor affinity for radioligand was higher in the white vastus than in the mixed fiber and slow-twitch muscles (Kd = 3.5 +/- 0.4 pM for white vastus versus 6.8 +/- 0.8 and 6.4 +/- 1.1 pM in gastrocnemius and soleus, respectively; both p less than 0.01 versus white vastus). Disparities in Bmax among muscles were due entirely to differences in receptor densities of skeletal muscle cells as shown autoradiographically. Furthermore, variations in Bmax of the three skeletal muscles were directly related to percentage of type I fibers (r = 0.99; p less than 0.001), which had a beta-receptor density that was approximately 4.5-fold greater than in superficially located type IIb fibers, 3.2-fold greater than in intermediate depth type IIb fibers, and 2.0-fold greater than in type IIa fibers. In contrast, grain densities of resistance arterioles were similar regardless of surrounding skeletal muscle fiber type composition. However, resistance arterioles were 2.5- and 6.1-fold more numerous in the slow-twitch soleus than in the gastrocnemius and superficial white vastus, respectively (all differences p less than 0.01). We conclude that beta-receptor density of rat hindquarter skeletal muscles is directly proportional to percentage of slow-twitch fibers, while receptor affinity for antagonist is higher in fast-twitch than in slow-twitch or mixed fiber muscles.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases↗

Low-pressure-sensitive baroreceptor fibers recorded from rabbit carotid sinus nerves.

Activity was recorded from physiologically identified baroreceptor or chemoreceptor fibers in carotid sinus nerves of urethane-anesthetized spontaneously breathing rabbits. A carotid sinus area was vascularly isolated so that carotid sinus pressure and perfusion medium (Locke's solution or rabbit blood) could be controlled. The cervical sympathetic, vagus, and aortic depressor nerves were bilaterally cut to eliminate vagal and cardiopulmonary reflexes. Baroreceptor fibers could be divided into two groups: fibers with a mean firing threshold of 47.6 +/- 1.9 mm Hg and no activity below this threshold (37 fibers) and fibers that were active at low intrasinus pressures (18.1 +/- 2.2 impulses/sec at an intrasinus pressure of 0 mm Hg). The baroreceptor fibers that were spontaneously active at low pressures were also chemically sensitive: discharge rate was increased by 5-hydroxytryptamine (10 fibers, p less than 0.01), nicotine (10 fibers, p less than 0.01), or hypercapnia (13 fibers, p less than 0.001). The activity of baroreceptor fibers with a clear pressure threshold was usually decreased by hypercapnia (26 of 27 fibers, from 18.8 +/- 3.1 to 13.2 +/- 3.9 impulses/sec). Chemoreceptor fibers failed to respond to intrasinus pressure changes from 0 to 100 mm Hg (n = 25 fibers, p greater than 0.5) but were sensitive to chemical changes, as expected. Thus, there is a subset of baroreceptor fibers that, under certain conditions, is spontaneously active at very low intrasinus pressures and responds to changes in the chemical milieu.

Action Potentials↗

Surface roughness enhances upward migration of bacteria on polymer fibers above liquid cultures.

Monofilament polypropylene (PP) fibers, very similar to fibers that have been used as monofilament tailstrings of interuterine contraceptive devices, were suspended vertically in bacterial liquid monocultures so that a portion of a fiber extended above the liquid surface. In some cases these highly oriented, cold drawn fibers were abraded prior to insertion in the cultures in order to produce surface roughness characterized by axial channels and protruding microfibrils that partially peeled from the fiber surface thereby forming the channels. Extent of migration on a fiber was assessed by aseptically cutting it into small segments, followed by culturing each segment on agar containing growth medium. Such assessment of the PP fibers after 48 h of incubation in the cultures revealed upward migration of Eschericia coli, Pseudomonas aeruginosa, and Staphylococcus aureus over significantly longer distances on the pre-roughened fibers than on those not so pre-treated. Mean measured distances of migration during 48 h were: for E. coli 2.7+/-0.6 mm on roughened fibers (n = 16) and 0.4+/-0.7 mm on fibers not roughened (n = 17); for S. aureus 9.0+/-4.3 mm on roughened fibers (n = 13) and 0.2+/-0.3 mm on fibers not roughened (n = 14); for P. aeruginosa 8.5+/-3.7 mm on roughened fibers (n = 26) and 0.2+/-0.5 mm on fibers not roughened (n = 5). Although no statistically significant (95% confidence level) difference could be discerned between the migration distances of S. aureus and P. aeruginosa, each of these species migrated a greater distance on the PP than did E. coli. The migrations observed are attributed predominantly to wicking of the liquid cultures upward in the axial grooves developed on the surface of the PP by the eruption and peeling of microfibrils from the surface. Surface tension of the growth medium was significantly lower than that of water and its contact angle on PP was less than 90 deg, thereby indicating a tendency to wet the PP. Bacterial growth in the medium further reduced its contact angle on PP, thereby indicating an even greater tendency to wet PP after such growth.

Animals↗

The role of clearance and dissolution in determining the durability or biopersistence of mineral fibers.

It is generally accepted that to cause pulmonary disease, mineral fibers must be relatively long and thin but also able to remain in the lung for long periods. This "biopersistence" of fibers is limited by two main mechanisms of fiber clearance: removal by macrophages after phagocytosis and, for some fibers, by actual dissolution. The relative importance of these mechanisms has not been properly evaluated for any type of fiber and will certainly vary with mineral type. The efficiency of macrophage clearance is greatest with short fibers (< 5 microns long) and is reduced as fibers get longer. Fibers > 50 microns long cannot be cleared by macrophages and for some mineral types they may remain in the lung permanently. Others may fracture into shorter lengths, perhaps aided by chemical dissolution, and thus become susceptible to macrophage clearance. However, for a number of areas relating to fiber removal from the lung parenchyma detailed information is still needed: Do dusts differ in their ability to attract macrophages and stimulate these cells to phagocytosis? Following dust uptake what controls the movement of macrophages? Some may penetrate to the interstitium, some phagocytosing fibers in interstitial sites may migrate back to the alveolar space. Some move to the mucociliary escalator and some to the lymphatics. Some, most importantly, move to the pleura. Fibers are found and phagocytosed in the interstitium during the early stages of disease development, but with time many fibers appear isolated in areas of fibrous tissue. Are such fibers subsequently ignored or can they reenter the disease process after years of isolation? Finally, can phagocytosis by macrophages effect dissolution of fibers?(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗