Psychophysical Investigations into the Role of Low-Threshold C Fibres in Non-Painful Affective Processing and Pain Modulation

We recently showed that C low-threshold mechanoreceptors (CLTMRs) contribute to touch-evoked pain (allodynia) during experimental muscle pain. Conversely, in absence of ongoing pain, the activation of CLTMRs has been shown to correlate with a diffuse sensation of pleasant touch. In this study, we evaluated (1) the primary afferent fibre types contributing to positive (pleasant) and negative (unpleasant) affective touch and (2) the effects of tactile stimuli on tonic muscle pain by varying affective attributes and frequency parameters. Psychophysical observations were made in 10 healthy participants. Two types of test stimuli were applied: stroking stimulus using velvet or sandpaper at speeds of 0.1, 1.0 and 10.0 cm/s; focal vibrotactile stimulus at low (20 Hz) or high (200 Hz) frequency. These stimuli were applied in the normal condition (i.e. no experimental pain) and following the induction of muscle pain by infusing hypertonic saline (5%) into the tibialis anterior muscle. These observations were repeated following the conduction block of myelinated fibres by compression of sciatic nerve. In absence of muscle pain, all participants reliably linked velvet-stroking to pleasantness and sandpaper-stroking to unpleasantness (no pain). Likewise, low-frequency vibration was linked to pleasantness and high-frequency vibration to unpleasantness. During muscle pain, the application of previously pleasant stimuli resulted in overall pain relief, whereas the application of previously unpleasant stimuli resulted in overall pain intensification. These effects were significant, reproducible and persisted following the blockade of myelinated fibres. Taken together, these findings suggest the role of low-threshold C fibres in affective and pain processing. Furthermore, these observations suggest that temporal coding need not be limited to discriminative aspects of tactile processing, but may contribute to affective attributes, which in turn predispose individual responses towards excitatory or inhibitory modulation of pain.     

Allodynic skin in post-herpetic neuralgia: histological correlates

Most post-herpetic neuralgia (PHN) patients suffer from tactile allodynia (pain evoked by lightly touching the skin) and it is frequently the dominant clinical manifestation. The pathophysiology of tactile allodynia in PHN patients is poorly understood and this is one of the major limits to the development of appropriate therapies. Epidermal nerve fibres (ENFs) are free nerve endings of small-diameter A-delta and C primary afferents, which can easily be assessed by neurodiagnostic skin biopsy (NSB). The aim of this study was to establish the correlation between the residual epidermal innervation of the allodynic skin and the intensity of tactile allodynia in that area. Twenty-five patients (13 males and 12 females) with PHN were enrolled. Eighteen patients had PHN in the thoracic dermatome, four in the cervical, two in the trigeminal and one in the lumbar. The severity of allodynia evoked by a paintbrush was graded according to an eleven-point numerical scale. A skin biopsy was obtained from the maximal allodynia area and from the contralateral skin. Nerve fibres were labelled with indirect immunofluorescence. Results showed that epidermal innervation was lower in the allodynic skin than in the contralateral skin, although there was great variability among patients. There was no correlation between severity of allodynia and epidermal innervation of the PHN skin. In conclusion, the present study further indicates peripheral nervous system involvement in PHN but does not support a direct correlation between epidermal innervation changes and tactile allodynia.     

The effects of heat-induced pain on the detectability,discriminability, and sensation magnitude of vibrotactile stimuli

The effects of heat-induced pain on absolute thresholds, sensation magnitudes and amplitude-difference thresholds were measured at 10 and 100 Hz. Consistent with previous results, heat-induced pain elevated the absolute thresholds by approximately 8.0 dB and lessened the magnitudes of tactile sensations during pain as compared to the non-painful condition. In contrast to these effects, the discriminability of change in the intensity of the vibrotactile stimuli was unaffected by the presence of pain indicating that the effect of pain on tactile sensations is more likely due to sensory rather than cognitive processes (i.e., attention) and that the mechanisms underlying tactile sensitivity as compared to discriminability are different.     

The effects of heat-induced pain on the detectability, discriminability, and sensation magnitude of vibrotactile stimuli

The effects of heat-induced pain on absolute thresholds, sensation magnitudes and amplitude-difference thresholds were measured at 10 and 100 Hz. Consistent with previous results, heat-induced pain elevated the absolute thresholds by approximately 8.0 dB and lessened the magnitudes of tactile sensations during pain as compared to the non-painful condition. In contrast to these effects, the discriminability of change in the intensity of the vibrotactile stimuli was unaffected by the presence of pain indicating that the effect of pain on tactile sensations is more likely due to sensory rather than cognitive processes (i.e., attention) and that the mechanisms underlying tactile sensitivity as compared to discriminability are different.     

Monoaminergic and cholinergic nerve fibres in the human dental pulp

Summary Distribution of cholinesterase-containing nerve fibres was studied in 15 human dental pulps by the thiocholine method. Falk's fluorescent method was used to demonstrate catecholamines (8 dental pulps).Cholinesterases were localized partly in the subodontoblastic plexus sending out fine branches towards odontoblasts, and partly in the nerve fibres attached to the blood vessel walls. These fibres in contrast to those of the subodontoblastic plexus were finer and showed fine varicosities.Monoaminergic terminals were localized mainly along blood vessel walls, however, some fibres having no relation to the blood vessels were also found.Cholinesterase-containing nerve fibres in the periphery of the pulp are considered to be sensitive nerve fibres originating from n.V. Distribution of cholinesterase-containing nerve fibres and monoaminergic terminals along the blood vessel walls indicates that the blood vessels in the human dental pulp might be under both parasympathetic and sympathetic control.     

Phosphatidylinositol, a phospholipid of ice-nucleating bacteria.

The nature of the phospholipids of the various bacteria that have ice nucleation activity in supercooled water has been determined. The seven bacteria studied included Pseudomonas syringae, Erwinia herbicola, three Escherichia coli K-12 strains that are phenotypically Ice+ because they contain plasmids with different amounts of either P. syringae or E. herbicola cloned DNA, and two E. coli K-12 strains without cloned ice gene DNA. All five Ice+ bacterial strains contained small amounts (0.1 to 1.0% of the total phospholipids) of phosphatidylinositol (PI), a phospholipid not previously detected in E. coli, Pseudomonas, or Erwinia species. The Ice- E. coli strains also contained trace level of PI that amounted to 2 to 30% of the level found in the Ice+ E. coli strains. Extracts of Ice+ strains contained low but measurable activities of PI synthase, while the activities in Ice- strains amounted to only 8 to 12% or less of that found in extracts of Ice+ bacteria. The functioning of the ice gene apparently increased both the PI synthase activity and the PI content of Ice+ strains from low endogenous levels. The relative ice nucleation activity at -4 degrees C or above (class A nucleation activity) of all Ice+ strains was found to be proportional to their PI content. The addition of myo-inositol (5 x 10(-4) M) to synthetic culture media increased the class A nucleation activity of both Ice+ E. coli strains and P. syringae up to sevenfold but had no stimulating effect on ice nucleation at lower temperatures (class B and class C nucleation activities). If these cells after fusion with PI vesicles were incubated with an energy source, the class A nucleation activity increased 70-fold over that present before fusion. These results indicate that PI plays an important role in ice nucleation at warm temperatures and is a likely precursor or component of the class A structure.     

Ice crystals formed in tissues during cryosurgery. I. Light microscopy

The size and distribution of ice crystals formed during cryosurgical procedures in intact animals are not clear. In the present experiment oral mucosa was frozen in situ by means of a surface applied cold probe and was excised and freeze substituted while in the frozen state. It was shown that the form of the frozen tissue was preserved during this procedure and the area frozen was divisible into a zone representing the central part of the lesion and a peripheral zone separating this from normal tissue. Ice crystals within the body of the lesion were intracellular in location but varied somewhat in size. Ice crystals in the boundary zone appeared to be intracellular in the epithelium and both intra- and extracellular in the muscle fibres.It is suggested that the intracellular crystals in the body of the frozen area result in cell death while the extracellular ice in the boundary zone results in a less predictable response.     

The effects of stimulus location on the gating of touch by heat- and cold-induced pain

The influence of heat- and cold-induced pain on tactile sensitivity, a "touch gate", was measured under conditions in which the location of the noxious stimuli was varied with respect to the tactile stimulus applied to the thenar eminence of humans. Vibrotactile thresholds were measured in the absence of pain and during administration of a painful stimulus, with the stimulus frequencies selected to activate independently the four psychophysical channels hypothesized to exist in human glabrous skin. Heat-induced pain produced by spatially co-localizing the noxious stimuli with the tactile stimuli was found, on average, to elevate threshold amplitude by 2.2 times (6.7 dB). Co-localized, cold-induced pain raised the average thresholds by about 1.5 times (3.6 dB). Heat-induced pain presented contralaterally produced no change in vibrotactile sensitivity indicating that the effect is probably not due to attentional mechanisms. Ipsilateral heat-induced pain caused an elevation in tactile thresholds even when the noxious and non-noxious stimuli were not co-localized, and the effect may seem to require that the painful stimulus be within the somatosensory region defined possibly in terms of dermatomal organization. Thus the effect is probably related to somatotopic organization and is not peripherally mediated. A brief discussion as to the possible locus of the touch gate within the nervous system is also given.     

Safety zone as a barrier to root-shoot ice propagation

Winter wheat, rye and triticale were sown at a depth of 12 cm to promote internode elongation. The root-shoot junction was separated by an internode about 4.5 cm long. The rate of ice propagation was measured separately across the root-internode and crown-internode at –4°C. Ice nucleation in seminal roots was experimentally induced by ice nucleating bacteria. The rate of movement of the ice front was measured by differential thermal analysis. The highest rate of ice propagation occurred in the internode. The rate of ice propagation across the root-internode section was slower than in the internode alone, but differences exist in the various genotypes. The rate of ice propagation between the seminal roots and the internode did not correlate with the number of roots with unsafe vessels, as detected by the pressurized air method. The rate of ice propagation between the internode and the crown was approximately one third of the rate in the internode. The discontinuity in internode-crown section reached nearly 100% as was observed by the pressurized air method. Anatomical studies supported the discontinuity of vessels evaluated by the pressurized air method. Experimental data support the view that the barriers in root-internode and in internode-crown section reduced the rate of ice propagation but these barriers are not correlated with either cold hardiness or number of safe roots.Abbreviations AN adventitious nodal roots - AS adventitious seminal roots - SS secondary seminal roots     

The effects of stimulus location on the gating of touch by heat- and cold-induced pain

The influence of heat- and cold-induced pain on tactile sensitivity, a "touch gate", was measured under conditions in which the location of the noxious stimuli was varied with respect to the tactile stimulus applied to the thenar eminence of humans. Vibrotactile thresholds were measured in the absence of pain and during administration of a painful stimulus, with the stimulus frequencies selected to activate independently the four psychophysical channels hypothesized to exist in human glabrous skin. Heat-induced pain produced by spatially co-localizing the noxious stimuli with the tactile stimuli was found, on average, to elevate threshold amplitude by 2.2 times (6.7 dB). Co-localized, cold-induced pain raised the average thresholds by about 1.5 times (3.6 dB). Heat-induced pain presented contralaterally produced no change in vibrotactile sensitivity indicating that the effect is probably not due to attentional mechanisms. Ipsilateral heat-induced pain caused an elevation in tactile thresholds even when the noxious and non-noxious stimuli were not co-localized, and the effect may seem to require that the painful stimulus be within the somatosensory region defined possibly in terms of dermatomal organization. Thus the effect is probably related to somatotopic organization and is not peripherally mediated. A brief discussion as to the possible locus of the touch gate within the nervous system is also given.     

Ice crystal patterns in artificial gels of extracellular matrix macromolecules after quick-freezing and freeze-substitution

Artificial gels, composed of collagen with or without hyaluronate (HA), a glycosaminoglycan (GAG), and chondroitin sulfate (CS), were prepared and quick-frozen for the purpose of studying the influence of composition and concentration on ice patterns. Dilute gels were spread on coverslips, plunged into a slush of 30% isopentane/70% propane (-185 degrees C), freeze-substituted, and examined by phase-contrast microscopy. Ice patterns were revealed as "ice cavities" in the gel after freeze-substitution. Ice morphology in the gels was gel-type-specific, suggesting that composition in dilute gels can influence ice pattern formation. Crystallization patterns reflecting high, intermediate, and low rates of freezing were observed in all gel types. Intermediate freezing in differentiating gel-type-specific ice patterns. Gels which included hyaluronate (HA) and chondroitin sulfate (CS) altered the ice crystal pattern commonly observed in collagen gels. Ice structure in collagen gels consisted predominantly of long, parallel crystals in the herringbone pattern. Ice crystals separated gel into thin, unbranched fibers with a primary spacing of approximately 2 microns. Ice morphology in HA gels formed a mosaic consisting of packets of ice crystals. Contiguous packets were often oriented at right angles to each other. Periodic crossbridges interconnect primary gel fibers of HA gels and interrupt the lengthwise growth of ice crystals. Smooth beads were visible on primary strands in HA gels frozen at intermediate velocities. The addition of CS to collagen gels resulted in formation of randomly oriented ice crystals in gels frozen at intermediate rates. CS has little influence on ice morphology at low freezing velocities. Primary strands in CS gels were decorated with rough-surfaced, osmiophilic aggregates.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ice crystals formed in tissue during cryosurgery. II. Electron microscopy

Tissues frozen by means of a cryosurgical probe have been examined by electron microscopy following techniques designed to preserve the ice crystal spaces.Ice crystals appeared similar whether tissues were quenched or not following cryosurgery and the various techniques of dehydration resulted in similar ice crystal architecture.Ice crystal spaces in the area deep to the freezing probe were intracellular both in epithelium and muscle although in the muscle zone some fibers contained large and others small crystal spaces. It is suggested that this might be due to variations in the local blood supply.At the periphery of the frozen area ice crystals were usually extracellular producing gross distortion of the cells which, however, retained intracellular structural integrity. These results are consistent with the belief of many workers that intracellular ice is lethal while extracellular ice is not, but no evidence of penetration of cell membrane by ice crystals was seen.     

The great Cenozoic ice sheet

Ocean-floor sediments contain a record of Quaternary continental glaciations in which global cooling prevails over a 100,000 year cycle and the intensity of the last glacial maximum is as strong as any of the earlier glaciations. Yet, the areal extent of Quaternary glacial drift in North America and Eurasia during the last glaciation is much less extensive than during several earlier glaciations. In an attempt to explore this problem, two versions of a Great Cenozoic Ice Sheet were reconstructed. The minimum version transformed a frozen bed beneath high interior ice domes and along southern margins during the last glaciation into a thawed bed during more extensive earlier glaciations. This allowed thinner ice to cover a larger area with little change in ice volume, thereby satisfying both the continental and oceanic records of Quaternary glaciation. The maximum version extended frozen basal conditions beneath high interior ice domes and along southern margins, so that a greater areal extent could only be accommodated by thicker ice. A greater ice volume is required in this case, and would seem to put the Great Cenozoic Ice Sheet in the late Tertiary, where some evidence for global cooling on a 400,000 year cycle exists and might provide the longer timespan needed to produce a larger ice sheet.     

Environmental Predictors of Ice Seal Presence in the Bering Sea

Ice seals overwintering in the Bering Sea are challenged with foraging, finding mates, and maintaining breathing holes in a dark and ice covered environment. Due to the difficulty of studying these species in their natural environment, very little is known about how the seals navigate under ice. Here we identify specific environmental parameters, including components of the ambient background sound, that are predictive of ice seal presence in the Bering Sea. Multi-year mooring deployments provided synoptic time series of acoustic and oceanographic parameters from which environmental parameters predictive of species presence were identified through a series of mixed models. Ice cover and 10 kHz sound level were significant predictors of seal presence, with 40 kHz sound and prey presence (combined with ice cover) as potential predictors as well. Ice seal presence showed a strong positive correlation with ice cover and a negative association with 10 kHz environmental sound. On average, there was a 20–30 dB difference between sound levels during solid ice conditions compared to open water or melting conditions, providing a salient acoustic gradient between open water and solid ice conditions by which ice seals could orient. By constantly assessing the acoustic environment associated with the seasonal ice movement in the Bering Sea, it is possible that ice seals could utilize aspects of the soundscape to gauge their safe distance to open water or the ice edge by orienting in the direction of higher sound levels indicative of open water, especially in the frequency range above 1 kHz. In rapidly changing Arctic and sub-Arctic environments, the seasonal ice conditions and soundscapes are likely to change which may impact the ability of animals using ice presence and cues to successfully function during the winter breeding season.     

Nerve supply and distribution of cholinesterase activity in the external nose of the mole

Summary Nerve supply and the distribution of cholinesterase activity were studied in the skin of the external nose of seven moles using a simplified Bielschowsky-Gross silver method and Koelle's histochemical technique.The sensory units of the mole's nose or the organs of Eimer are surrounded by blood sinuses which facilitate their movements during mechanical stimulation. All nerve fibres of the plexus deep to the basal cell layer of Eimer's organ ultimately become intra-epidermal endings. Contrary to the findings of earlier investigators, Merkel's discs, Pacinian corpuscles and Ruffini corpuscles have not been observed at the base of Eimer's organ. In the superficial layer of the plexus, the Schwann sheath cells increase in number, undergo modification and give a positive cholinesterase reaction.It is suggested that the organ of Eimer, the specialised nerve plexus deep to it and the surrounding blood sinus together constitute the touch receptor on a similar principle of transmission by leverage as in the tactile hair or the intermediate ridge of the papillary ridge.The role of the intra-epidermal nerve endings of the mole's nose as tactile receptors is disputed. A suggestion is made that tnese nerves may constitute pain and temperature receptors and that several modalities of sensation may be carried to the brain along one and the same medullated axon.We gratefully acknowledge the technical assistance of Miss Jill Hocknell. Our thanks are also due to Mr. C. J. Duncan and the staff of the Photography Department for their aid with the photographic work. We are particularly grateful to Mr. D. Burgess of the Ministry of Agriculture and Fisheries for kindly supplying us with live moles. One of the authors (N.C.) acknowledges an equipment grant from the Royal Society.     

Ice Nucleation Activity in Fusarium acuminatum and Fusarium avenaceum

Twenty fungal genera, including 14 Fusarium species, were examined for ice nucleation activity at −5.0°C, and this activity was found only in Fusarium acuminatum and Fusarium avenaceum. This characteristic is unique to these two species. Ice nucleation activity of F. avenaceum was compared with ice nucleation activity of a Pseudomonas sp. strain. Cumulative nucleus spectra are similar for both microorganisms, while the maximum temperatures of ice nucleation were −2.5°C for F. avenaceum and −1.0°C for the bacteria. Ice nucleation activity of F. avenaceum was stable at pH levels from 1 to 13 and tolerated temperature treatments up to 60°C, suggesting that these ice nuclei are more similar to lichen ice nuclei than to bacterial ones. Ice nuclei of F. avenaceum, unlike bacterial ice nuclei, pass through a 0.22-μm-pore-size filter. Fusarial nuclei share some characteristics with the so-called leaf-derived nuclei with which they might be identified: they are cell free and stable up to 60°C, and they are found in the same kinds of environment. Highly stable ice nuclei produced by fast-growing microorganisms have potential applications in biotechnology. This is the first report of ice nucleation activity in free-living fungi.     

Adult spinal opioid receptor μ1 expression after incision is altered by early life repetitive tactile and noxious procedures in rats

Clinical and experimental data suggests that noxious stimulation at critical stages of development results in long‐term changes on nociceptive processing in later life. Here, we use an established, well‐documented rat model of repetitive noxious procedures closely mimicking the clinical situation in the NICU. In order to understand molecular changes underlying the long‐term consequences of repetitive stimulation of the developing nociceptive system the present study aims to analyze the presence of the µ‐opioid‐receptor‐1 (OPRM1). Neonatal rats received either four needle pricks per day in the left hind‐paw from postnatal day 0–7 as a model of procedural pain in infancy. Control pups were handled in the same way but were instead tactile stimulated, or were left undisturbed. At the age of 8 weeks, all animals received an ipsilateral hind‐paw incision as a model for post‐operative pain, and mechanical sensitivity was tested at multiple time‐points. Before, and 1 or 5 days post‐incision, spinal cord tissue was collected for immunostaining of opioid receptor OPRM1. Semi‐quantitative immunocytochemical analysis of superficial laminae in lumbar spinal dorsal horn revealed that: (1) early life repetitive tactile or noxious procedures do not alter baseline levels of OPRM1 staining intensity and (2) early life repetitive tactile or noxious procedures lead to a decrease in OPRM1 staining intensity 5 days after incision in adulthood compared to undisturbed controls. We conclude that early life repetitive tactile or noxious procedures affect the intensity of OPRM1‐immunoreactivity in the lumbar superficial spinal cord dorsal horn after adulthood injury, without affecting baseline intensity. © 2018 The Authors. Developmental Neurobiology Published by Wiley Periodicals, Inc. Develop Neurobiol 78: 417–426, 2018     

中药熏蒸联合推拿治疗腰椎间盘突出症的临床效果

目的：探讨中药熏蒸联合推拿治疗腰椎间盘突出症的临床效果。方法：选择2013 年12 月到2015 年1 月我院收治的112 例 腰椎间盘突出症患者为研究对象,根据治疗方法的不同分为研究组和对照组。对照组给予常规推拿治疗措施,研究组给予常规推 拿措施联合中药熏蒸治疗。比较两组患者的住院时间,下肢放射痛、麻木、腰痛的发生率及临床疗效。结果：研究组患者的住院时 间及下肢放射痛、麻木、腰痛的发生率均较对照组明显缩短或降低,其治疗总有效率为92.9%,显著高于对照组,均有统计学差异 (P＜0.05)。结论：中药熏蒸联合推拿方法治疗腰椎间盘突出症的临床效果优于单一推拿治疗措施,可有效改善患者的临床症状,缩 短疗程,提高治疗有效率,值得临床推广使用。     

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.     

Welfare aspects of theriogenology: investigating alternatives to electroejaculation of bulls

Evaluation of the breeding soundness of bulls is an important management tool. Electroejaculation has been a reliable method of obtaining a semen sample for the purpose of evaluating breeding soundness, but is considered by some to be inhumane on the grounds that it is painful. This paper provides a review of studies conducted to find ways to both measure, as well as lessen, pain associated with electroejaculation, and to explore alternatives to electroejaculation in bulls. Changes in heart rate, serum cortisol, serum progesterone, relative aversion, and degrees of vocalization, struggling and lying down have been used to assess the pain associated with electroejaculation. Transrectal massage and artificial vaginas, and oxytocin and cloprostenol have been investigated as alternatives to, and facilitators of electroejaculation, respectively. Epidural, intravenous and topical anesthetics have been used to ameliorate the pain associated with electroejaculation. Serum progesterone and degrees of vocalization are useful for measuring the pain associated with electroejaculation in bulls. Transrectal massage and artificial vaginas are not as efficacious as electroejaculation for obtaining a semen sample and drugs used to facilitate or decrease pain associated with electroejaculation have not been efficacious enough to warrant use. Transrectal massage of the ampullae may be of some use as an alternative to electroejaculation in docile bulls and may be also be used to decrease the duration of subsequent electroejaculation. Pain associated with electroejaculation may be influenced by operator technique; therefore, operators of electroejaculator equipment must strive to apply electrical stimulation as gently as possible.