Cold Stimulation Research Articles

Transplantation of neural stem cells preconditioned with high‑mobility group box 1 facilitates functional recovery after spinal cord injury in rats.

Spinal cord injury (SCI) is a devastating disorder that often results in temporary and/or permanent functional impairment below the injured level. To date, few satisfactory therapeutic strategies are available to treat SCI. Hence, exploring novel strategies for SCI is an essential public health concern. Cell transplantation therapy, which is associated with neuroprotection, immunomodulation, axon regeneration, neuronal relay formation and myelin regeneration, provides a promising therapeutic strategy for SCI. The neuronal stem cell (NSC) preconditioning method is an emerging approach, which facilitates NSC survival and neuronal differentiation after implantation. The aim of the present study was to develop a feasible candidate for cell-based therapy following SCI in rats and to investigate the role of high mobility group box-1 (HMGB1) in NSC activation. The results of the present study showed that transplantation of NSCs, preconditioned with 1 ng/ml HMGB1, facilitated functional improvement of injured spinal cords, as indicated by Basso, Beattie and Bresnahan mean scores, mechanical hypersensitivity and cold stimulation. Meanwhile, the histological examination of hematoxylin and eosin staining indicated that engraftment of HMGB1-preconditioned NSCs resulted in decreased atrophy of the injured spinal cord. Meanwhile, the transplantation of HMGB1-preconditioned NSCs resulted in an increased number of functional Nissl bodies in neurons, as detected by Nissl staining, and an increase in the number of βIII-tubulin+ cells in the epicenter of injured spinal cords in rats with SCI. In addition, the results also demonstrated that 1 ng/ml HMGB1 promoted the differentiation of NSCs into neurons, and that the ERK signaling pathway played an important role in this process. In conclusion, the present data indicated that the preconditioning strategy with 1 ng/ml HMGB1 may present a feasible candidate for cell-based therapy following SCI in rats, which may enlarge the scope of HMGB1 in NSC activation.

Intact vitamin A transport is critical for cold-mediated adipose tissue browning and thermogenesis

ObjectiveTransformation of white into brown fat (“browning”) reduces obesity in many preclinical models and holds great promise as a therapeutic concept in metabolic disease. Vitamin A metabolites (retinoids) have been linked to thermogenic programming of adipose tissue; however, the physiologic importance of systemic retinoid transport for adipose tissue browning and adaptive thermogenesis is unknown. MethodsWe performed cold exposure studies in mice and humans and used a genetic model of defective vitamin A transport, the retinol binding protein deficient (Rbp−/-) mouse, to study the effects of cooling on systemic vitamin A and the relevance of intact retinoid transport on cold-induced adipose tissue browning. ResultsWe show that cold stimulation in mice and humans leads to an increase in circulating retinol and its plasma transporter, Rbp. In Rbp−/- mice, thermogenic programming of adipocytes and oxidative mitochondrial function are dramatically impaired in subcutaneous white fat, which renders Rbp−/- mice more cold-sensitive. In contrast, retinol stimulation in primary human adipocytes promotes thermogenic gene expression and mitochondrial respiration. In humans, cold-mediated retinol increase is associated with a shift in oxidative substrate metabolism suggestive of higher lipid utilisation. ConclusionsSystemic vitamin A levels are regulated by cold exposure in mice and humans, and intact retinoid transport is essential for cold-induced adipose tissue browning and adaptive thermogenesis.

Non-Invasive Analysis of Actinic Keratosis before and after Topical Treatment Using a Cold Stimulation and Near-Infrared Spectroscopy.

Background and objectives: The possible evolution of actinic keratoses (AKs) into invasive squamous cell carcinomas (SCC) makes their treatment and monitoring essential. AKs are typically monitored before and after treatment only through a visual analysis, lacking a quantitative measure to determine treatment effectiveness. Near-infrared spectroscopy (NIRS) is a non-invasive measure of the relative change of oxy-hemoglobin and deoxy-hemoglobin (O2Hb and HHb) in tissues. The aim of our study is to determine if a time and frequency analysis of the NIRS signals acquired from the skin lesion before and after a topical treatment can highlight quantitative differences between the AK skin lesion area. Materials and Methods: The NIRS signals were acquired from the skin lesions of twenty-two patients, with the same acquisition protocol: baseline signals, application of an ice pack near the lesion, removal of ice pack and acquisition of vascular recovery. We calculated 18 features from the NIRS signals, and we applied multivariate analysis of variance (MANOVA) to compare differences between the NIRS signals acquired before and after the therapy. Results: The MANOVA showed that the features computed on the NIRS signals before and after treatment could be considered as two statistically separate groups, after the ice pack removal. Conclusions: Overall, the NIRS technique with the cold stimulation may be useful to support non-invasive and quantitative lesion analysis and regression after a treatment. The results provide a baseline from which to further study skin lesions and the effects of various treatments.

Conduction velocity of the cold spinal pathway in healthy humans.

We aimed to investigate the conduction velocity of the cold spinal pathway in healthy humans. Using a cold stimulator consisting of micro-Peltier elements that was able to produce steep cooling ramps up to -300°C/s, we recorded cold-evoked potentials after stimulation of the dorsal midline at C5, T2, T6 and T10 vertebral levels and calculated the conduction velocity of the cold spinal pathway. In all participants, we used laser stimulation to deliver painful heat (Aδ-fibres-mediated) and warm (C-fibres-mediated) stimuli to the same sites in order to compare the conduction velocity of the cold spinal pathway with that of the nociceptive and warm spinal pathways. Cold stimulation evoked large-amplitude vertex potentials from all stimulation sites. The mean conduction velocity of the cold spinal pathway was 12.0m/s, which did not differ from that of the nociceptive spinal pathway (10.5m/s). The mean conduction velocity of the warm spinal pathway was 2.0m/s. This study provides previously unreported findings regarding cold spinal pathway conduction velocity in humans that may be useful in the assessment of spinal cord lesions as well as in intraoperative monitoring during spinal surgery. This neurophysiological study provides previously unreported findings on cold spinal pathway conduction velocity in healthy humans. Cold-evoked potentials may represent an alternative to laser-evoked potential recording, useful to assess spinothalamic tract in patients with spinal cord lesions and monitor patients during spinal surgery.

Intestinal tract and parenteral multi-organ sequential pathological injury caused by necrotizing enterocolitis

BackgroundTo explore the relationship between the pathological changes of the colon, terminal ileum, lung, liver and kidney, and the changes of Bax, PCNA and PAF in a rat model of NEC.MethodsOne hundred and forty neonatal SD rats were randomly divided into NEC group and control group (70 in each group). NEC group was given hypoxia, cold stimulation and artificial feeding twice a day for 3 consecutive days. The control group was only fed normally. After modeling, From the 1st day to the 7th day, 10 rats were sampled in each group for pathological examination of colon, terminal ileum, lung, liver and kidney tissue. The levels of Bax, PCNA and PAF were investigated by immunohistochemistry.ResultsCompared with the normal group, in the NEC group, on the 1st day, the colon, terminal ileum, lung, liver and kidney showed inflammatory damage. On the 5th day, the inflammatory injury was reduced. The inflammation disappeared on the 7th day. There were differences in the time of apoptosis in the intestine. In the intestine, the proliferation of PCNA was weak at first and then strong. Bax in liver and kidney showed marked apoptosis and apoptosis time increased in the lung. The expression of PCNA increased in lung, liver and kidney, and the expression of PAF increased in lung and liver.ConclusionsNEC can lead to secondary injury of different degrees in colon, terminal ileum, lung, liver and kidney, and the degree and time of injury and repair were different. In general, organ repair played a leading role on the 4th day after modeling.

Effect of wrist-ankle acupuncture on the expression of glutamate and NMDA receptor of the spinal dorsal horn in rats with neuropathic pain

To observe the effect of wrist-ankle acupuncture (WA) stimulation at "R4"- "R5" - "R6" on the expression of glutamate (Glu) and phosphorylated protein NMDAR1(p-NMDAR1) of the spinal dorsal horn in spared nerve injury (SNI) rats, so as to explore its mechanism underlying improvement of SNI. A total of 36 SD rats were randomly divi-ded into sham operation, model and WA groups, with 12 rats in each group. The SNI procedure comprised an axotomy and ligation of the tibial and common peroneal nerves leaving the sural nerve intact. Rats of the WA group were treated by acupuncture at "R4"-"R5"-"R6" points from the 5th day to the 14th day after modeling. The mechanical pain thresholds were measured before and 5, 10 and 14 d after SNI, respectively. The cold allodynia was dectected by Acetone solution dropped onto the lateral plantar surface of the paw. Glu content and p-NMDAR1 expression of spinal dorsal horn were detected by 1H-MRS, ELISA and immunohistochemistry Methods. Compared with the sham operation group, the mechanical pain threshold of the model group was significantly decreased (P<0.01), the duration of cold stimulation foot contraction was increased (P<0.01), and the Glu content and p-NMDAR1 expression in the spinal dorsal horn were significantly increased (P<0.05, P<0.01). After WA intervention, the mechanical pain threshold was significantly increased (P<0.01), the duration of cold stimulation was significantly shortened (P<0.01), and Glu content and p-NMDAR1 protein expression of spinal dorsal horn were decreased significantly (P<0.05, P<0.01) in the WA group compared with the model group. WA can reduce pain sensitivity in rats with neuropathic pain, possibly by inhibiting the expression of Glu and p-NMDAR1 in the spinal dorsal horn.

Effects of Intermittent Mild Cold Stimulation on mRNA Expression of Immunoglobulins, Cytokines, and Toll-Like Receptors in the Small Intestine of Broilers

Simple SummaryCold stress has been associated with adverse effects on health and welfare of broilers. Whilst several studies have shown that long-term sustained and mild cold stimulation can improve immune function, little is known of the effects of intermittent cold stimulation on immune modulation in broilers. In this study, broilers were submitted to cold stimulation of 3 °C below than the usual rearing temperature during 3 and 6 h every two days during 43 days to explore its effect on the intestinal immunity. The findings confirm that appropriate mild cold stimulation has an overall positive influence on the intestinal immunity of broilers. The mild cold stimulation tested in this study is cost-effective and likely enhances overall health of broilers.Appropriate cold stimulation can improve immune function and stress tolerance in broilers. In order to investigate the effect of intermittent mild cold stimulation on the intestinal immunity of broilers, 240 healthy one-day-old Ross 308 chickens were randomly divided into three groups: the control group (CC) housed in climatic chambers under usual rearing ambient temperature with a gradual 3.5 °C decrease per week; group II (C3) and group III (C6) to which cold stimulation at 3 °C below the temperature used in CC was applied every two days for 3 and 6 h, respectively, from day 15 to 35, and at the same temperature used in CC from day 35 to 43. The mRNA expression levels of immunoglobulins (IgA and IgG), cytokines (IL2, IL6, IL8, IL17, and IFNγ), and Toll-like receptors (TLR2, TLR4, TLR5, TLR7, and TLR21) were investigated in duodenum, jejunum, and ileum tissue samples on days 22, 29, 35, and 43. From day 15 to 35, mRNA expression of IL2 and IFNγ was increased in the intestine of broilers. After one week of cold stimulation on day 43, mRNA levels of immunoglobulins, cytokines, and Toll-like receptors (TLRs) stabilized. Collectively, the findings indicate that cold stimulation at 3 °C below the usual rearing temperature had a positive impact on intestinal immunity of broilers.

Abstract 5593: Identification and characterization of an unusual monocyte subpopulation

Abstract Personalized immunotherapy for cancer has become an effective treatment choice in certain oncology indications. To successfully develop such therapeutic candidates requires a deep understanding of the biology including the interaction between a variety of hematopoietic cells, mesenchymal cells, and the microenvironment of bone marrow and peripheral blood. Flow cytometry offers unparalleled advantages in immunophenotyping, functional analysis, and receptor occupancy determination. For example, flow cytometry is capable of simultaneously analyzing millions of leucocytes and identifying populations of regulatory T cells, cytotoxic T cells, B cells, NK cells, monocytes, macrophages, and dendritic cells with multiple surface and intracellular biomarkers. Because of the diversity of clinical samples, variable protocols, and storage conditions, new challenges arise during the development and flow cytometric analysis of immunotherapeutic drug candidates. One of these challenges is the observation, upon flow cytometric analysis, of an unexpected pattern of immune cells. For example, monocytes are one of the important components in the immune system, but they are also readily modulated during an immune response. In our clinical studies, we found that some peripheral whole blood samples exhibit an extra population paralleling normal monocytes during flow cytometric analysis. To correctly categorize these cells, we applied a panel of 18 fluorochrome conjugated monoclonal antibodies, such as CD34, CD11c, CD1b, to define this uncharacterized cluster. Peripheral whole blood samples were maintained at ambient temperature as well as refrigerated at 4°C. The monocyte subpopulation was most evident in samples maintained at refrigerated temperature. We used a gating strategy to separate the two clusters, normal monocytes and the additional high forward scatter (FSC) monocyte subpopulation in FCS Express analysis software. Using custom designed data analysis templates, we performed side-by-side comparison of these two populations. The results indicated that the two clusters demonstrated a similar profile in this panel. In conclusion, we found that this extra cluster phenotyped as an altered subpopulation of monocytes, and was most evident following cold stimulation. The significance of this monocyte subpopulation remains unknown at the current time. The importance of panel design in flow cytometry was highlighted in this work. The results and subsequent bioinformatics proved informative in characterizing this unusual subpopulation by profiling an array of cellular biomarkers. Citation Format: Chengsen Xue, Joanne Cuomo, Lucie-Emily Rows, Kai Chen, Chrinstina D. Swenson, Thomas W. Mc Closkey. Identification and characterization of an unusual monocyte subpopulation [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5593.

Balneotherapy and hydrotherapy in chronic respiratory disease.

Chronic respiratory diseases (CRD) belong to major noncommunicable diseases (NCD) targeted by World Health Organization (WHO) NCD Action Plan and United Nations (UN) Sustainable Development Goal (STG) 3.4 to achieve 30% decline of mortality by the year 2030. Strong evidence is now available in the literature for therapeutic aquatic exercise interventions in improving health status of chronic obstructive pulmonary diseases (COPD) patients. However, gym-based exercises can be difficult for patients with COPD who are mainly elderly and often have co-morbidities—such as severe arthritis and obesity—which may impair their ability to exercise at an adequate intensity. Besides improving respiratory function and health status in COPD, exercise in water helps overcoming patient’s fears and promote socialization, contrasting the risk of depression, which is a major condition often associated with long term COPD condition. Susceptibility to respiratory infections plays a role in exacerbations of COPD. Sulphur-rich water inhalations improve muco-ciliary clearance, reduce inflammatory cytokines production and inflammatory mucosal infiltration, reduce elastase secretion by neutrophils, preserving elastic properties of pulmonary interstitium and thus facilitating expectoration. Repeated cold water stimulations in COPD also reduce frequency of infections. Finally, sauna bathing reduces the risk of pneumonia. On the other side, hydrotherapy/balneotherapy also help obesity control, which is one of the most difficult NCD risk factors to modify and consequently is an important component of the WHO preventive strategy to achieve STG 3.4. Along with high prevalence and mortality, CRD cause increasing pharmaceutical and hospital costs. In this perspective, Health Resort Medicine should not be ignored as a resource in the WHO NCD strategy and Universal Health Coverage, providing a multi-stakeholder platform (including the network of health resorts and their facilities) able to give a real help to the achievement of UN goal STG 3.4 by the year 2030.

Endomorphin analog exhibited superiority in alleviating neuropathic hyperalgesia via weak activation of NMDA receptors

Morphine is a key drug for the treatment of pain but its side effects limit its clinical application. MEL-0614, an endomorphin-1 analog, has fewer side effects than morphine in addition to its powerful analgesic effect. In this study, we measured the effect of morphine and MEL-0614 on hyperalgesia (7days) and neuropathic allodynia (14days) after thermal, mechanical, and cold stimulation. We found that after four and eight consecutive days of intrathecal administration (1, 3, and 10nmol), morphine induced severe hyperalgesia and neuropathic allodynia, respectively. MEL-0614 did not induce hyperalgesia at low doses (1 and 3nmol) and had a mitigating effect on morphine-induced neuropathic exacerbations in spared nerve injury mice. Hyperalgesia was blocked by Dynorphin A (1-17) antibody but not by an opioid receptor antagonist. To explore the reasons for the different results of morphine and MEL-0614, we used quantitative PCR and immunofluorescence to explore the effects of both on N-methyl-D-aspartate (NMDA) receptor subtype 2B (NR2B), microglia marker iba-1, and inflammatory mediators. After 8days of consecutive administration, morphine (10nmol) promoted an increase in the number of NR2B, iba-1, and inflammatory mediators in the spinal cord of mice. MEL-0614 (10nmol) had no significant effect on these factors, and after co-administration with morphine, the expression of NR2B, iba-1, and inflammatory mediators was lower than that with morphine injection alone. Our research showed the advantage of MEL-0614 in terms of hyperalgesia and neuropathic allodynia, which may provide clinical relief of hyperalgesia and neuropathic allodynia caused by morphine.

Pain Processing in Elite and High-Level Athletes Compared to Non-athletes.

BackgroundPrevious studies shows that elite and high-level athletes possess consistently higher pain tolerance to ischemic and cold pain stimulation compared to recreationally active. However, the data previously obtained within this field is sparse and with low consistency.PurposeThe aim of the present study was to examine the difference in pain perception between elite and high-level endurance athletes (cross country skiers and runners), elite soccer players and non-athletes, as well to explore the impact of psychological factors on pain processing.MethodsSeventy one healthy volunteers (33 females and 38 males) participated in the study. Soccer players (n = 17), cross country skiers (n = 12), and long-distance runners (n = 3) formed the athlete group, with 39 non-athletes as controls. Big-five personality traits, fear of pain and Grit (perseverance and passion for long-term goals) were measured prior to induction of experimental pain. Pain threshold and intensity was induced by a PC-controlled heat thermode and measured by a computerized visual analog scale. Pain tolerance was measured by the cold pressor test (CPT).ResultsElite and high-level athletes had increased pain tolerance, higher heat pain thresholds, and reported lower pain intensity to thermal stimulation. Endurance athletes (cross country skiers and long-distance runners) had better tolerance for cold pain compared to both soccer-players and non-athletes. Furthermore, endurance athletes reported lower pain intensity compared to non-athletes, whereas both endurance athletes and soccer players had higher heat pain thresholds compared to non-athletes. Fear of Pain was the only psychological trait that had an impact on all pain measures.ConclusionThe present findings suggest that sports with long durations of physically intense activity, leveling aerobic capacity, are associated with increased ability to tolerate pain and that the amount of training hours has an impact on this tolerance. However, the small sample size implies that the results from this study should be interpreted with caution.

Cold pain hypersensitivity predicts trajectories of pain and disability after low back surgery: a prospective cohort study.

Improving the ability to predict persistent pain after spine surgery would allow identification of patients at risk and guide treatment decisions. Quantitative sensory tests (QST) are measures of altered pain processes, but in our previous study, preoperative QST did not predict pain and disability at single time-points. Trajectory analysis accounts for time-dependent patterns. We hypothesized that QST predict trajectories of pain and disability during 1 year after low back surgery. We performed a trajectory analysis on the cohort of our previous study (n = 141). Baseline QST included electrical, pressure, heat, and cold stimulation of the low back and lower extremity, temporal summation, and conditioned pain modulation. Pain intensity and Oswestry Disability Index were measured before, and 2, 6, and 12 months after surgery. Bivariate trajectories for pain and disability were computed using group-based trajectory models. Multivariable regressions were used to identify QST as predictors of trajectory groups, with sociodemographic, psychological, and clinical characteristics as covariates. Cold pain hypersensitivity at the leg, not being married, and long pain duration independently predicted worse recovery (complete-to-incomplete, incomplete-to-no recovery). Cold pain hypersensitivity increased the odds for worse recovery by 3.8 (95% confidence intervals 1.8-8.0, P < 0.001) and 3.0 (1.3-7.0, P = 0.012) in the univariable and multivariable analyses, respectively. Trajectory analysis, but not analysis at single time-points, identified cold pain hypersensitivity as strong predictor of worse recovery, supporting altered pain processes as predisposing factor for persisting pain and disability, and a broader use of trajectory analysis. Assessment of cold pain sensitivity may be a clinically applicable, prognostic test.

Effect of Intermittent and Mild Cold Stimulation on the Immune Function of Bursa in Broilers.

Simple SummaryCold stress has an adverse effect on immune, antioxidant, and neuroendocrine systems as well as growth performance of broilers. Much attention was focused on how to improve the cold resistance of animals. However, it is not clear how intermittent and mild cold stimulation regulate the immune function. Therefore, the present study was designed to investigate the effect of intermittent and mild cold stimulation on the immune function of bursa in broilers. Our results indicated that the intermittent and mild cold stimulation (IMCS) could help broilers adapt to the low ambient temperature and maintain homeostasis by modulating the production of Toll-like receptors (TLRs), cytokines, and avian β-defensins (AvBDs) in the bursa of broilers. We expect to find a way to promote the establishment of cold adaptation through repeated intermittent cold trainings.Cold stress causes growth performance to decrease and increases production costs. Cold adaptation can enhance immune function and alleviate the negative impact caused by the stress condition. The study investigated the effect of intermittent and mild cold stimulation on the immune function of the bursa of Fabricius in broilers. A total of 400 healthy one-day-old broilers were divided into the control group (CC) and cold stimulation (CS) groups. The CC group was raised at a conventional raising temperature of broilers, while the CS groups were raised at 3°C below the temperature of the CC for three-, four-, five-, or six-hour periods at one-day intervals from 15 to 35 days of age (D35), denoted CS3, CS4, CS5, and CS6, respectively. Subsequently, they were raised at 20°C from 36 to 49 days of age (D49). The expression levels of TLRs, cytokines, and AvBDs were determined to access the immune function of bursa in broilers. After 21-day IMCS (at D36), the expression levels of TLR1, TLR15 and TLR21, interleukin (IL)-8, and interferon (IFN)-γ, as well as AvBD8 in CS groups, were lower than those in CC (p < 0.05). The expression levels of TLR3, TLR4 and TLR7, were decreased in the CS3, CS5, and CS6 groups (p < 0.05), but there were no significant differences in both the CC and CS4 groups (p > 0.05). When the IMCS ended for 14 days (at D49), the expression levels of TLR2, TLR3, TLR5, TLR7, TLR15, and TLR21, and IL-8, as well as AvBD2, AvBD4 and AvBD7 in CS groups, were lower than those in CC (p < 0.05). In addition to CS4, the expression levels of TLR1, IFN-γ, and AvBD8 in CS3, CS5, and CS6 were still lower than those in CC (p < 0.05). We concluded that the intermittent and mild cold stimulation could regulate immunoreaction by modulating the production of TLRs, cytokines, and AvBDs in the bursa, which could help broilers adapt to low ambient temperature and maintain homeostasis.

Heightened affective response to perturbation of respiratory but not pain signals in eating, mood, and anxiety disorders.

Several studies have recently suggested that an abnormal processing of respiratory interoceptive and nociceptive (painful) stimuli may contribute to eating disorder (ED) pathophysiology. Mood and anxiety disorders (MA) are also characterized by abnormal respiratory symptoms, and show substantial comorbidity with ED. However, no studies have examined both respiratory and pain processing simultaneously within ED and MA. The present study systematically evaluated responses to perturbations of respiratory and nociceptive signals across the levels of physiology, behavior, and symptom report in a transdiagnostic ED sample (n = 51) that was individually matched to MA individuals (n = 51) and healthy comparisons (HC; n = 51). Participants underwent an inspiratory breath-holding challenge as a probe of respiratory interoception and a cold pressor challenge as a probe of pain processing. We expected both clinical groups to report greater stress and fear in response to respiratory and nociceptive perturbation than HCs, in the absence of differential physiological and behavioral responses. During breath-holding, both the ED and MA groups reported significantly more stress, feelings of suffocation, and suffocation fear than HC, with the ED group reporting the most severe symptoms. Moreover, anxiety sensitivity was related to suffocation fear only in the ED group. The heightened affective responses in the current study occurred in the absence of group differences in behavioral (breath hold duration, cold pressor duration) and physiological (end-tidal carbon dioxide, end-tidal oxygen, heart rate, skin conductance) responses. Against our expectations, there were no group differences in the response to cold pain stimulation. A matched-subgroup analysis focusing on individuals with anorexia nervosa (n = 30) produced similar results. These findings underscore the presence of abnormal respiratory interoception in MA and suggest that hyperreactivity to respiratory signals may be a potentially overlooked clinical feature of ED.

Lasting modulation of human cortical swallowing motor pathways following thermal tongue stimulation.

Thermal tactile oropharyngeal stimulation has been clinically used to facilitate swallowing initiation in dysphagic patients. We previously demonstrated that thermal stimulation applied to the oral cavity provokes an immediate excitability in pharyngeal motor cortex. The aim of the current study was to investigate whether thermal stimulation can produce longer lasting effects on the corticopharyngeal neural pathway. Healthy volunteers (n=8/12) underwent baseline pharyngeal motor evoked potential (PMEP) measurements evoked by transcranial magnetic stimulation. In the first experiment, subjects received thermal stimulation alternating 30seconds of 15 and 36°C applied to the tongue surface for either 10minutes, 5minutes, or sham. In the second experiment, one of three intermittent thermal stimulus patterns was delivered: cold (alternating 30seconds of 15 and 36°C), warm (continuous 36°C), or hot (alternating 30seconds of 45 and 36°C) for 10minutes. In both experiments, PMEP were remeasured every 15minutes up to 60minutes following thermal stimulation. Repeated measures ANOVA for each stimulus time in the first experiment showed a significant increased change in PMEP amplitude at 30minutes following only 10-minute stimulation compared with sham (P<.05). In the second experiment, we found that cold stimulation was more effective than the other stimulation (P<.05) at increasing PMEP amplitudes. Ten-minute cold stimulation on the tongue can induce a delayed (30minutes) increase in pharyngeal cortical excitability, providing a clinically useful therapeutic window for its application in dysphagic patients.

The α2,3-selective potentiators of GABAA receptors, KRM-II-81 and MP-III-80, produce anxiolytic-like effects and block chemotherapy-induced hyperalgesia in mice without tolerance development

Opiate analgesics are one of the treatment options for severe chronic pain, including late-stage cancer, chronic back pain and other disorders. The recent resurgence in opioid overdose has highlighted the serious need for alternative medicines for pain management. While a role for potentiators of α2/3-containing GABAA receptors in the modulation of pain has been known for several years, advancements in this area required data from selective compounds. KRM-II-81(5-(8-ethynyl-6-(pyridin-2-yl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepin-3- yl)oxazole) and analogs selectively potentiate GABAA receptors containing α2/3 subunits and have recently been shown to attenuate pain behaviors in several acute and chronic pain models in rodents. The present study was designed to ascertain whether KRM-II-81 and the structural analog MP-III-80 (3-ethyl-5-(8-ethynyl-6-(pyridin-2-yl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepin-3-yl)-1,2,4-oxadiazole) would block chemotherapeutic agent paclitaxel-induced pain in male, C57BL/6 mice. Both compounds significantly inhibited pain behaviors evoked by cold and tactile stimulation in paclitaxel-treated mice as did the neuropathic pain drug gabapentin. Subchronic dosing for 22 days with KRM-II-81 and MP-III-80 demonstrated enduring analgesic efficacy without tolerance development, while the effects of gabapentin showed evidence of tolerance development. KRM-II-81 and MP-III-80 also decreased marble-burying behavior in this mouse strain as did the anxiolytic drug chlordiazepoxide. In contrast to KRM-II-81 and MP-III-80, chlordiazepoxide had motor-impairing effects at anxiolytic-like doses. The data add to the literature documenting that these selective potentiators of α2/3-containing GABAA receptors are effective in a host of animal models used to detect novel analgesic drugs. The anxiolytic-like efficacy of these compounds fits well with the comorbidity of anxiety in patients with chronic pain and cancer.

Normalized skin conductance level could differentiate physical pain stimuli from other sympathetic stimuli

Skin conductance monitoring is one of the promising methods for objectively evaluating pain. However, skin conductance might possibly increase in response to sympathetic stimulation other than pain. In this study, we aimed to test whether skin conductance monitoring can distinguish physical pain stimulation (heat, mechanical and cold stimulation) from other sympathetic stimuli (stimulation by noise and painful images). Twenty-three healthy volunteers participated in this prospective observational study. The number of fluctuations in skin conductance (NFSC) and normalized skin conductance level (nSCL) were measured and compared with pain scores on a self-reported pain scale (numerical pain scale [NPS]). Both NFSC and nSCL increased during mechanical stimulation. Further, nSCL, but not NFSC, well reflected heat stimulus intensity, suggesting its ability to quantitatively evaluate pain. nSCLs during physical pain stimulation were greater than those during other sympathetic stimulations. However, NFSC was not able to completely distinguish between the stimuli. These results suggest that nSCL could better differentiate physical pain stimuli from other sympathetic stimuli than NFSC. In comparisons between subjective and objective pain assessment in the same individual, nSCL correlated better with NPS score, indicating the possibility of being able to monitor the transition of pain. Monitoring changes in skin conductance using nSCL might be useful for objectively detecting physical pain.

The effects of season change and fasting on Brown adipose tissue FDG-PET in mice

It is widely reported that BAT is more frequently observed in patients during the winter season, and its activities could vary significantly under different conditions. However, whether this phenomenon is entirely caused by low temperature or other factors is not very clear. In this study, we tried to explore the seasonal fluctuation of FDG-PET BAT using mouse models that were from the same genetic breed and raised in a well-controlled environment. We also compared these variations with the effects of fasting and cold stimulation on BAT activities in these mice. In overnight fasted mice, the FDG-PET BAT was the highest in standardized uptake value (SUV) in the winter season. The values were much lower in all other seasons, especially in the summer. Compared to regular feeding, overnight fasting reduced BAT SUV, and refeeding after fasting could fully recover BAT activities. Fasted mice also did not respond to cold environment stimulation. After refeeding, their BAT thermogenic activities became normal. These results suggest that BAT FDG-PET SUV measurements vary significantly with the season and highlight the importance of taking into account the seasonal effect and fasting status in BAT evaluation studies using FDG-PET imaging.

Antinociceptive Effects of Emodin on CFA-Induced Inflammatory Pain in Rats

The effect of emodin on complete Freund’s adjuvant (CFA)-induced inflammatory pain in rats and its potential molecular mechanism was investigated. For this, a rat model of inflammatory pain induced by CFA was established and rats were treated with emodin by intraperitoneal injection. The pain threshold was evaluated by the von Frey, thermo hyperalgesia, and cold plate tests. The mRNA expression of transient receptor potential channel ankyrin type-1 ( Trpa1) and transient receptor potential vanilloid 1 ( Trpv1) was detected by quantitative reverse transcription polymerase chain reaction, and the level of inflammatory cytokines was determined by enzyme-linked immunosorbent assay. The mechanical and thermal pain thresholds of CFA-treated rats were significantly lower than those of the control rats, while the paw withdrawal responses in response to cold stimulation were higher than that of the control group. Emodin treatment significantly improved CFA-induced hyperalgesia. Further results showed that emodin inhibits the upregulation of Trpa1 and Trpv1 mRNA expression in the dorsal root ganglion (DRG) of rats with inflammatory pain compared with the control group. Emodin also significantly reduced the levels of tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6) in the serum of rats with inflammatory pain. Thus, emodin may inhibit hyperalgesia induced by inflammatory stimulation by downregulating the mRNA expression of Trpa1 and Trpv1 in DRG neurons and reducing the levels of TNF-α, IL-1β, and IL-6.

Influence of Intermittent Cold Stimulations on CREB and Its Targeting Genes in Muscle: Investigations into Molecular Mechanisms of Local Cryotherapy.

Local cryotherapy is widely used as a treatment for sports-related skeletal muscle injuries. The molecular mechanisms are unknown. To clarify these mechanisms, we applied one to three 15-min cold stimulations at 4 °C to various cell lines (in vitro), the tibialis anterior (TA) muscle (ex vivo), and mouse limbs (in vivo). In the in vitro assay, cyclic AMP (cAMP) response element binding protein 1 (CREB1) was markedly phosphorylated (p-CREB1), and the CREB-binding protein (CBP) was recruited to p-CREB-1 in response to two or three cold stimulations. In a reporter assay with the cAMP-responsive element, the signals significantly increased after two to three cold stimulations at 4 °C. In the ex vivo study, CREB-targeting genes were significantly upregulated following two or three cold stimulations. The in vivo experiment disclosed that cold stimulation of a mouse limb for 9 days significantly increased mitochondrial DNA copy number and upregulated genes involved in mitochondrial biogenesis. The results suggest that local cryotherapy increases CREB transcription and upregulates CREB-targeting genes, in a manner dependent on cold stimulation frequency and duration. This information will inform further investigations into local cryotherapy as a treatment for sports-related skeletal muscle trauma.