Variables associated with corneal confocal microscopy parameters in healthy volunteers: implications for diabetic neuropathy screening

Diabet. Med. 29, e297-e303 (2012) ABSTRACT: Aim Corneal confocal microscopy is a promising screening method for diabetic neuropathy. Although much research in this field has been accomplished, we aimed to determine and confirm the known clinical and eyewear variables associated with the parameters of corneal confocal microscopy specifically in healthy volunteers, in particular associations with corneal nerve fibre length. Methods Clinical characteristics, electrophysiological examination and a general clinical eye history were collected from 64 healthy volunteers. Corneal confocal microscopy was performed to determine corneal nerve fibre length, corneal nerve branch density, corneal nerve fibre density and tortuosity coefficient. Univariate and multivariate linear regression analysis was used to determine clinical variables associated with corneal nerve fibre length parameters. Results We observed that corneal nerve fibre length has a broad distribution in healthy volunteers (18?±?4?mm/mm(2) , 95% confidence interval, 12.3-25.7?mm/mm(2) ). Multivariate regression analysis demonstrated that HbA(1c) was the only independent clinical factor to account for variations in corneal nerve fibre length, independent of age and status of contact lens wear. Conclusions This study does not provide convincing evidence that corneal nerve fibre length is independently associated with age or the wearing of contact lenses, and that these factors are therefore unlikely to hinder valid screening for polyneuropathies such as diabetic neuropathy. Furthermore, the strong inverse association of corneal nerve fibre length with glycaemic exposure may support the use of this parameter to detect subclinical pre-diabetic nerve injury.     

Morphology of Schmidt-Lanterman incisures of human shoulder plexus nerves in ontogenesis

The Schmidt-Lanterman incisures are formed in the phase of "loose" myelin at 15-17 weeks of prenatal development with the formation of nodes of Ranvier. The formation of mature Schmidt-Lanterman incisure is completed in the second foetis period. Schmidt-Lanterman incisures distribution in nerve fibres of young men depends on fibre diameter. In the nerves of elderly people the distribution of the incisures in the internode doesn't depend on a fibre diameter. In the nerves of the elderly men the number of incisures is twice as much as that in the young people. The incisural ultrastructure in human nerve is similar to its structure in other mammals.     

Annual variations of the NPYergic innervation of the pineal gland in the European hamster (Cricetus cricetus): a quantitative immunohistochemical study

A prominent innervation of the pineal gland of the European hamster with nerve fibres containing neuropeptide Y (NPY) and tyrosine hydroxylase (TH) was demonstrated by means of immunohistochemistry. Nearly all the TH- and NPY-immunoreactive nerve fibres in the superficial pineal gland disappeared after bilateral superior cervical ganglionectomy, showing that the majority of NPY- and TH-immunoreactive nerve fibres belonged to the sympathetic nervous system. Since, in the European hamster, preliminary studies of the NPY-fibre density in the pineal gland had indicated seasonal changes, the density of NPY-immunoreactive nerve fibre profiles was ascertained in the superficial pineal gland in a series of animals between the first part of November and late April. The highest density of NPY-immunoreactive nerve fibre profiles was observed during midwinter. On the other hand, during the same period of the year, the number of sympathetic TH-immunoreactive sympathetic nerve fibre profiles did not exhibit seasonal variation, nor did substitution of testosterone, during the sexually inactive period, affect the density of NPY-containing nerve fibres in the gland. Our results show the presence of a testosterone-independent annual variation in the content of NPY in the sympathetic nerve fibres innervating the pineal gland of the European hamster. This variation can be correlated with the changes in the daily pattern of melatonin production observed by others in the same species at this period of the year.     

Tapering of human nerve fibres

To determine the tapering of human nerve fibres, rostral and caudal root pieces of cauda equina nerve roots were removed and nerve fibre diameter distributions were constructed for 4 myelin sheath thickness ranges for the two sites, and compared with each other. The reduction of the group diameter in the different alpha-motoneuron groups was 0.2 % per 13 cm. Accounting for systematic errors, there may be even less tapering. An identified single nerve fibre showed no tapering. Further, there is indication that gamma-motoneurons, preganglionic sympathetic and parasympathetic fibres and skin afferents also reduce their fibre diameter by 0.2 % per 13 cm or less. Consequently, a nerve fibre with a diameter of 10 microm would be reduced to approximately 9.8 microm at 1m from the cell soma. Preganglionic parasympathetic fibres were found to be represented in roots S1 to S5. At similar distances from the spinal cord, the mean diameter of ventral root alpha1-motoneuron (FF) axons increased from the thoracic towards the lumbo-sacral region before decreasing again in the lower sacral region. Usually no alpha1-motoneuron axons were found in S5 roots. The diameter distribution of unmyelinated nerve fibres of a ventral S5 root showed three peaks at 0.25, 0.95 and 1.2 microm. The unmyelinated fibres with diameters around 0.25 microm may represent parasympathetic fibres. In six selected areas of the ventral S5 root, 6.6 times more unmyelinated nerve fibres than myelinated fibres were found on the average.     

Histochemical demonstration of some dehydrogenases and NAD-H diaphorase in cat Pacinian Corpuscles

Summary Using histochemical methods for G-6-P-DH, NAD-HD, M-DH, S-DH, L-DH and G-DH the author has examined the participation and distribution of these enzymes in Pacinian Corpuscles of male cats. The reactions for G-6-P-DH, L-DH, M-DH and NAD-HD show similarity regarding the intensity and the character of intrareceptoral distribution. The nerve fibre and its ending show the highest activity. Three other zones might be differentiated round them. The first zone situated in close proximity to the axon coincides with the inner core of the Corpuscles and shows higher activity than the others (second and third zones). The activity of the latter ones diminishes towards the periphery of the receptor.The reaction for S-DH is strongly expressed at the nerve fibre and its ending and weaker at the inner core of the Corpuscles.The reaction for G-DH is moderately expressed in the inner core whereas the nerve fibre and the outer core show the feeble activity.The author discusses the obtained results in relation to the necessity of energy and the ways it is obtained for the excitation of the axonal membrane.     

Effect of stimulus (postsynaptic current) shape on fibre excitation.

Effects of variation of the stimulus pulse shape on the excitation of a nonmyelinated nerve fibre were studied using a mathematical model based on the Hodgkin-Huxley equations. Efficiency of smoothly changing pulses was compared with that of rectangular pulses. For pulses shorter than the time to excitation, the rate of the stimulus rise did not determine the ability of a smoothly changing pulse to excite the fibre. For a given stimulus duration, the main factor was the pulse area or the charge delivered by the pulse. The strength-duration curve for smoothly changing pulses was a nonmonotonic function, in contrast to the curve for rectangular pulses. The dependence of latency on changes in the pulse area was non-linear. It would be nonmonotonic when the pulse area variation were due to the stimulus duration or the stimulus rise duration. More that one propagating intracellular action potential (IAP) could arise upon fibre activation by a long smoothly changing threshold stimulus. Upon activation of relatively short fibres the IAP could arise not at the site of the smoothly changing stimulus injection. The rectangular pulses of long duration were more efficient than the corresponding smoothly changing ones. Irrespective of the shape, the pulses whose duration at the foot is 1-2 ms, are more suitable for a prolonged threshold fibre activation.     

Comparative aspects of muscle fibre size and succinic dehydrogenase distribution in the longissimus dorsi muscle of several species of East African mammals

The distribution of succinic dehydrogenase enzyme activity was investigated in frozen sections of longissimus dorsi muscle taken from several species of East African game animal (giraffe, hartebeest, wildebeest, oryx, gerenuk and dik-dik) as well as local zebu cattle. Muscle fibres were classified as red (high succinic dehydrogenase activity), white (low activity) or intermediate. The mean diameter and percentage distribution of each fibre type were noted as well as the overall mean muscle fibre diameter (MFD) for each species. The diameters of red muscle fibres were found to be between 54 and 62% of the diameters of the white muscle fibres for all species with MFD differences between species being up to over 100%. The variation in MFD was found to be significantly and positively correlated with live weight, when zebu values were omitted. It was found that the variation in MFD was significantly related to the diameter of muscle fibre types and not to the percent distribution of these fibre types.     

Measures of variability in age-structured populations

An expression for the entropy of a population was derived in Demetrius (1974) by using a variational principle argument. This entropy measure is precisely the information content of the distribution in the ages of reproducing individuals in a stationary population. This paper introduces another expression for the entropy by considering the variation in the ages at which offspring will be produced by newborn individuals.The relation between these two measures of entropy and their biological significance are discussed.     

Calbindin neurons of the guinea-pig small intestine: quantitative analysis of their numbers and projections

Summary The distribution of nerve cells with immunoreactivity for the calcium-binding protein, calbindin, has been studied in the small intestine of the guinea-pig, and the projections of these neurons have been analysed by tracing their processes and by examining the consequences of nerve lesions. The immunoreactive neurons were numerous in the myenteric ganglia; there were 3500±100 reactive nerve cells per cm² of undistended intestine, which is 30% of all nerve cells. In contrast, reactive nerve cells were extremely rare in submucous ganglia. The myenteric nerve cells were oval in outline and gave rise to several long processes; this morphology corresponds to Dogiel's type-II classification. Processes from the cell bodies were traced through the circular muscle in perforating nerve fibre bundles. Other processes ran circumferentially in the myenteric plexus. Removal of the myenteric plexus, allowing time for subsequent fibre degeneration, showed that reactive nerve fibres in the submucous ganglia and mucosa came from the myenteric cell bodies. Operations to sever longitudinal or circumferential pathways in the myenteric plexus indicated that most reactive nerve terminals in myenteric ganglia arise from myenteric cell bodies whose processes run circumferentially for 1.5 mm, on average. It is deduced that the calbindin-reactive neurons are multipolar sensory neurons, with the sensitive processes in the mucosa and with other processes innervating neurons of the myenteric plexus.     

Effect of ten different target tissues on nerve fibre outgrowth from rat sympathetic ganglia in organotypic co-cultures

Sympathetic thoracic chain ganglia of 3-day-old rats were cultured in collagen gel medium for 24 hours together with explants from heart atrium, liver, kidney, cornea, iris, lung, adrenal cortex, adrenal medulla, skeletal muscle, or vas deferens. The extent of nerve fibre growth was estimated by counting the number of fibres crossing each arc of a sector drawn in the ocular. The various tissues stimulated nerve fibre growth to distinctly different extents. The increase in the nerve fibre outgrowth induced by atrium and iris was statistically highly significant. Kidney, liver, vas deferens, lung, and adrenal cortex had, in that order, a decreasingly stimulatory influence on sympathetic chain ganglia. Yet they all caused a significant increase in nerve fibre growth. Skeletal muscle, cornea and adrenal medulla had no stimulatory effect. Since the significant effects of the tissue explants were abolished by antiserum to nerve growth factor (NGF), it is concluded that the observed effects were due to NGF produced by the explants. The only exception was vas deferens, the stimulatory action of which proved to be partially NGF-independent.     

Neurokinin A-like immunoreactivity in feline dental pulp: its distribution,origin and coexistence with substance P-like immunoreactivity

Summary The distribution and origin of neurokinin A (NKA)-like immunoreactivity were investigated in feline dental pulp by an indirect immunofluorescence method. NKA-containing nerve fibres with varicosities, which entered the dental pulp via apical foramen, were distributed throughout this tissue. Many NKA-containing nerve fibres were localized around blood vessels, but some were observed apart therefrom. At the odontoblastic layer, thin NKA-containing nerve fibres were observed running straight toward the pulp-predentinal border between odontoblasts. After inferior alveolar nerve section, all NKA-containing nerve fibres disappeared in the dental pulp, while the removal of the superior cervial ganglion resulted in no change in the distribution of these fibres. The correlation of NKA-like immunoreactivity and substance P (SP)-like immunoreactivity was also investigated by double-immunofluorescence technique. The distribution of NKA-containing nerve fibres was very similar to that of SP-containing nerve fibres; it appeared that all NKA-containing nerve fibres contained SP.     

Effect of foreign innervation on the androgen-sensitive levator ani muscle of the rat

Summary Cross-union of the tibial with the pudendal nerve innervating the androgen-sensitive levator ani (LA) muscle of male rats, results in reversal of the histochemical muscle fibre pattern concerning myofibrillar ATPase, succinate dehydrogenase and phosphorylase enzyme activities. The homogeneous muscle fibre pattern of the LA muscle is changed to a mosaic pattern of muscles normally innervated by the tibial nerve. The success of the hetero-reinnervation is shown by practically full recovery of muscle weight and of isometric twitch-contraction properties of the LA muscle. Castration of 2-months duration, i. e. lack of the male sex hormone, leads to marked atrophy but no change in histochemical muscle fibre pattern. Hetero-reinnervation of the LA muscle results in change of histochemical enzyme pattern even if the cross-union of nerves is performed after long periods of castration leading to very marked decrease of muscle fibre size. However, testosterone application alone after castration increases markedly muscle fibre size but does not lead to reversal of muscle fibre pattern. The myotropic hormonal influence on the target (LA) muscle is therefore primarily of myogenic origin and specificity of hormonal action is maintained even with a foreign nerve innervating the muscle. The experiments, thus, provide evidence for the differentiation of specific neural influences affecting muscle fibre pattern and hormonal influences in respect to the myotropic action of the sex hormone on the androgen-sensitive LA muscle.     

AUTORADIOGRAPHIC STUDY OF RNA IN NERVE FIBRES OF EMBRYONIC SENSORY GANGLIA CULTURED IN VITRO UNDER NGF STIMULATION

Abstract— The presence and distribution of RNA in nerve fibres was studied by autoradiographic detection of [³H]uridine incorporation in chick embryo sensory nerve cells cultured in vitro under the effect of the Nerve Growth Factor. The presence of RNA in these fibres, free from satellite cells, is demonstrated. The cellular origin of the fibre RNA is, at least in part, proved.     

Immunoreactive atrial and brain natriuretic peptides are co-localized in Purkinje fibres but not in the innervation of the bovine heart conduction system

Summary Recently, we observed that atrial natriuretic peptide (ANP) immunoreactivity was present in Purkinje fibres and nerve fibre varicosities in the conduction system of the bovine heart. In order to elucidate further the morphological correlation between natriuretic peptides and the conduction system, the distribution of brain natriuretic peptide (BNP) was examined. The different parts of the conduction system in the bovine heart were dissected out and processed for immunohistochemistry with antisera against BNP and ANP. BNP immunoreactivity was frequently observed in Purkinje fibres of the atrioventricular bundle, whereas only a few Purkinje fibres in the ventricular part of the conduction system showed immunoreaction. BNP immunoreactivity was detected in regions of the Purkinje fibres which also showed ANP immunoreactivity. BNP immunoreactivity was not observed in nerve fibre varicosities. Methodologically, a larger number of small BNP immunofluorescent granular structures was observed by using an elution-restaining technique instead of conventional immunohistochemistry. The present study shows that BNP and ANP immunoreactivities frequently occur in the atrioventricular bundle and that they are co-localized in Purkinje fibres, but not in nerve fibre varicosities, in the conduction system. As previously has been proposed for ANP, the present observations suggest that also BNP may act in an autocrine and/or paracrine way in the conduction cells.     

X-ray observations on a collagen fibril lattice structure in peripheral nerve

It has been known for more than two decades that peripheral nerve shows X-ray reflections other than those originating from the myelin sheath. These extra reflections are at small angles of diffraction and arise from a variation of electron density in the radial direction. X-ray diffraction studies since 1973 have identified these reflections as coming from a lattice structure of filaments, the filament axes are parallel to the nerve fibre axis. The present X-ray observations were obtained using a high-resolution X-ray camera and we conclude that these reflections arise from a collagen fibril lattice structure within peripheral nerve. Estimates of fibril radius and the separation distance between fibrils in intact rat, rabbit and frog sciatic nerves have been obtained using X-ray analysis.     

Implications to fleece evaluation derived from sources of variation contributing to cashmere fibre curvature

Cashmere fibre curvature (crimp) has important impact on the softness and quality of cashmere textiles, the efficiency of cashmere processing and cashmere production. This work was aimed to quantify the magnitude and direction of factors affecting cashmere fibre curvature, with data collected from 11 Australian commercial cashmere farms, using general linear model analysis. Nineteen parameters were recorded for 1244 goats. Following log transformation the best model for fibre curvature included farm, age, clean washing yield, mean fibre diameter, cashmere yield, fibre diameter standard deviation, and live weight and the interactions between these terms. The percentage variance accounted for was 71.7%. Mean fibre diameter alone accounted for 39% of the variation in fibre curvature and farm accounted for 49% of the variation. Cumulatively mean fibre diameter and farm accounted for 66.6% of the variation existing in fibre curvature. For the other terms, age added 2.2% and the other fibre measurements a further 2.9% to variation accounted for by the best model. Results suggest that within a farm, using cashmere fibre crimp frequency to estimate mean fibre diameter has a correlation of 0.72—provided the trained observers perform as well as the calibrated laboratory equipment. On the other hand, however, results indicate fibre curvature not to be a reasonable indicator of mean fibre diameter differences across farms. Farm-effects on fibre curvature are large and may explain the difficulties cashmere growers experience when they visit other farms to visually evaluate cashmere goats prior to purchase. This work indicated that heavier goats are likely to produce cashmere with a lower fibre curvature. As this relationship did not differ between farms, it is reasonable to conclude that all goats exhibit this phenotypic response. Using cashmere fibre curvature (crimp frequency) as a tool for changing mean fibre diameter or selecting homogenous batches of fibre for sale will be reasonably effective within a farm, but is not a reasonable indicator and predictor of mean fibre diameter differences between farms.     

Fluorescence anisotropy and myelin structure. II. Azimuth characteristic of surviving nerve fibers]

The dependence of fluorescence polarization of stained nerve fibres on the angle between the fibre axis and electrical vector of exciting light (azimuth characteristics) has been considered. Evidence is provided that the azimuth characteristics of stained nerve fibres depends on dye molecules adsorbed on the myelin sheath membranes. From the previous calculations it may be concluded that part of the dye molecules are oriented at a small angle to the geometrical axis of the nerve fibre.     

The astrocytes in the retina and optic nerve head of mammals: A special glia for the ganglion cell axons

Summary The neuroglia in the retina and the intraocular portion of the optic nerve of the monkey and cat has been examined by light and electron microscopy. In the retina two types of macroglial cells can be distinguished: 1) Müller cells, and 2) astrocytes. The bipolar radial glial cells of Müller penetrate the entire thickness of the retina and their basal processes align in the nerve fibre layer to form septa that fasciculate the axons of the ganglion cells. In contrast to the Müller cells, the retinal astrocytes are not homogeneously distributed throughout the retina; their number correlates with the thickness of the nerve fibre layer. The processes of the astrocytes are confined to the ganglion cell layer and to the nerve fibre layer. In the latter, the astrocytic processes run parallel to and between the axons of a given nerve fibre bundle. According to cytological criteria, the retinal astrocytes are protoplasmic. In the intraocular portion of the optic nerve, however, the astrocytes are fibrous and their processes run perpendicular to the axon bundles of the prelaminar portion of the optic nerve. Thus, because of their intimate morphological relationship to axons of the nerve fibre layer and the intraocular portion of the optic nerve, the astrocytes in the eye of the monkey and the cat may be considered as a special glia for the axons of ganglion cells.