Peripheral Cold Research Articles

KCNQ1 is an essential mediator of the sex-dependent perception of moderate cold temperatures

Low temperatures and cooling agents like menthol induce cold sensation by activating the peripheral cold receptors TRPM8 and TRPA1, cation channels belonging to the TRP channel family, while the reduction of potassium currents provides an additional and/or synergistic mechanism of cold sensation. Despite extensive studies over the past decades to identify the molecular receptors that mediate thermosensation, cold sensation is still not fully understood and many cold-sensitive peripheral neurons do not express the well-established cold sensor TRPM8. We found that the voltage-gated potassium channel KCNQ1 (Kv7.1), which is defective in cardiac LQT1 syndrome, is, in addition to its known function in the heart, a highly relevant and sex-specific sensor of moderately cold temperatures. We found that KCNQ1 is expressed in skin and dorsal root ganglion neurons, is sensitive to menthol and cooling agents, and is highly sensitive to moderately cold temperatures, in a temperature range at which TRPM8 is not thermosensitive. C-fiber recordings from KCNQ1-/- mice displayed altered action potential firing properties. Strikingly, only male KCNQ1-/- mice showed substantial deficits in cold avoidance at moderately cold temperatures, with a strength of the phenotype similar to that observed in TRPM8-/- animals. While sex-dependent differences in thermal sensitivity have been well documented in humans and mice, KCNQ1 is the first gene reported to play a role in sex-specific temperature sensation. Moreover, we propose that KCNQ1, together with TRPM8, is a key instrumentalist that orchestrates the range and intensity of cold sensation.

Cold Habituation Does Not Alter Thermal Responses or Manual Dexterity During Prolonged Cold Air Exposure

Background: Habituation is a pattern of cold acclimatization characterized by attenuated cold defense responses, including a reduction in peripheral vasoconstriction and subsequent increase in skin temperature (TSk) as well as a decrease in shivering thermogenesis. Habituation is achieved through repeated reductions in TSk, rather than deep body core temperature (TC), via exposure to prolonged cold air or brief cold-water immersion. Due to improvements in TSk, enhanced manual dexterity and peripheral cold injury risk reduction are purported benefits of cold habituation. Warfighters frequently operate in cold climates and experience decrements in manual dexterity and strength. These functional losses can negatively impact mission critical tasks and training exercises; however, the effectiveness of cold habituation as a potential countermeasure to alleviate the deleterious effects of prolonged cold exposure on peripheral temperatures, thermal comfort, and hand function is unclear. Purpose: To evaluate the influence of eight consecutive cold air exposures on the thermal (TC and TSk), perceptual (thermal comfort and sensation), and functional (manual dexterity and strength) responses to cold air. We hypothesized that TSk would be higher and perceptual responses and manual dexterity improved, following repeated cold exposure. Methods: Eight young adults (6 M/1 F/1 FTM; 21 ± 2 years; %BF 24.5 ± 7.0) underwent a 2-h cold air exposure (8 °C, ~50% RH) on 8 consecutive days wearing minimal clothing. On Day 0 (baseline exposure ~1 week prior) and Days 1 and 8 of repeated exposure, TC was recorded continuously via a telemetry pill used as a rectal suppository. Extremity (Thand and Tfinger) and mean weighted TSk (MWTSk) from 8 skin sites were recorded continuously. Thermal comfort (TC) and thermal sensation (TS) were recorded at 20 min intervals. Expired gases were collected at 20 min intervals and used to calculate metabolic heat production (MHP). Manual dexterity was assessed using the Purdue Pegboard and Minnesota Manual Dexterity Test (MMDT), and hand strength were measured before and at the end of cold exposure. Results: TC changed across time ( p < 0.05) but did not differ between days ( p > 0.05). MWTSk, Thand, and Tfinger decreased across time ( p < 0.05) but also did not differ between days ( p > 0.05). Mean and minimum whole-body TS and TC were not different between days ( p > 0.05). MHP increased across time ( p < 0.05) but did not differ between days ( p > 0.05). Purdue Pegboard, MMDT task performance, and hand strength did not differ between days ( p > 0.05). Conclusion: Eight consecutive days of cold air exposure does not augment extremity skin temperature, thermal comfort, or manual performance during prolonged cold conditions. Supported by U.S. Army Medical Research and Development Command, Military Operational Medicine Research Program; author views not offcial US Army or DoD policy. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

A two-week exercise intervention improves cold symptoms and sleep condition in cold-sensitive women

BackgroundWe examined whether an aerobic exercise intervention in young women with cold sensitivity symptoms improves sleep quality and decreases cold complaints. Furthermore, we examined the association with increased foot skin temperature (Tsk) before falling asleep and decrease in sensitivity to cold in the brain.MethodsWe recruited 16 female adult volunteers who had cold sensitivity and were not engaged in daily exercise training, and they were divided into an exercise group (EXE) and a control group (CON). EXE was given a 2-week exercise intervention that consisted mainly of walking and jogging. Before and after the intervention, temperature sensation and body temperature parameters were measured just before bedtime; electroencephalogram measurements were taken during sleep; and subjective sleep surveys, including Oguri-Shirakawa-Azumi (OSA) sleep inventory (middle-aged and aged version) and visual analogue scale (VAS), were conducted immediately after waking up. All experiments were performed in the winter season.ResultsIn EXE, overall and foot warmth and comfort increased (p < 0.05) after the 2-week exercise intervention. The exercise intervention also decreased (p < 0.05) the scores for cold feeling in the fingertips, feet, and toes. In the OSA sleep inventory, factor IV (refreshing) and factor V (sleep length) were increased (p < 0.05) by the exercise intervention. Subjective sleep quality evaluated by VAS increased (p < 0.05) with exercise intervention. The exercise intervention in EXE shortened middle awake time after sleep onset (p < 0.05) and prolonged deep sleep length (p < 0.05). The exercise intervention increased (p < 0.05) alpha-wave power before sleep. In CON, all variables remained unchanged throughout the 2-week control period.ConclusionShort-term aerobic exercise alleviated peripheral extremity cold sensitivity symptoms and improved subjective sleep quality. Our findings suggest that these improvements were not due to increased Tsk at rest before bedtime but to decreased sensitivity to cold in the brain that was expressed as increased alpha activity.

IgM monoclonal gammopathy of undetermined significance: clinicopathologic features with and without IgM-related disorders.

A subset of patients with immunoglobulin M (IgM) monoclonal gammopathy of undetermined significance (MGUS) develop IgM-related disorders (IgM-RD) including peripheral neuropathy, cryoglobulinemia and/or cold agglutinin disease (CAD). We examined the clinical and bone marrow pathologic findings in 191 IgM MGUS patients (2016 World Health Oragnization criteria). Clonal plasma cells were identified in 41 of 171 (24%) cases by immunohistochemistry (IHC) and clonal B cells in 43 of 157 (27%). IgM-RD was identified in 82 (43%) cases, including peripheral neuropathy (n=67, 35%), cryoglobulinemia (n=21, 11%), and CAD (n=10, 5%). Cases of CAD showed distinctive features including lack of MYD88 mutations (P=0.048), supporting the concept of primary CAD as a distinct clinicopathologic disorder. Following exclusion of CAD, comparison of the remaining cases with (n=72) or without (n=109) IgM-RD showed IgM-RD to be more frequent in men than women (P=0.02) and to be more highly associated with MYD88 L265P (P=0.011). Cases with and without IgM-RD otherwise showed similar features including serum IgM concentrations, presence of lymphoid aggregates, clonal B cells by flow cytometry or clonal plasma cells by IHC. No differences were observed in overall survival between cases with and without IgM-RD. No cases in this series met criteria for plasma cell type IgM MGUS as defined in the 2022 International Consensus Classification of lymphoid neoplasms. These results show IgM-RD to be common in patients with IgM MGUS. While CAD shows distinctive features, the remaining cases of IgM-RD largely show pathologic findings similar to IgM MGUS without IgM-RD.

Chloride-dependent mechanisms of multimodal sensory discrimination and nociceptive sensitization in Drosophila.

Individual sensory neurons can be tuned to many stimuli, each driving unique, stimulus-relevant behaviors, and the ability of multimodal nociceptor neurons to discriminate between potentially harmful and innocuous stimuli is broadly important for organismal survival. Moreover, disruptions in the capacity to differentiate between noxious and innocuous stimuli can result in neuropathic pain. Drosophila larval class III (CIII) neurons are peripheral noxious cold nociceptors and innocuous touch mechanosensors; high levels of activation drive cold-evoked contraction (CT) behavior, while low levels of activation result in a suite of touch-associated behaviors. However, it is unknown what molecular factors underlie CIII multimodality. Here, we show that the TMEM16/anoctamins subdued and white walker (wwk; CG15270) are required for cold-evoked CT, but not for touch-associated behavior, indicating a conserved role for anoctamins in nociception. We also evidence that CIII neurons make use of atypical depolarizing chloride currents to encode cold, and that overexpression of ncc69-a fly homologue of NKCC1-results in phenotypes consistent with neuropathic sensitization, including behavioral sensitization and neuronal hyperexcitability, making Drosophila CIII neurons a candidate system for future studies of the basic mechanisms underlying neuropathic pain.

Hypothermia evoked by stimulation of medial preoptic nucleus protects the brain in a mouse model of ischaemia

Therapeutic hypothermia at 32-34 °C during or after cerebral ischaemia is neuroprotective. However, peripheral cold sensor-triggered hypothermia is ineffective and evokes vigorous counteractive shivering thermogenesis and complications that are difficult to tolerate in awake patients. Here, we show in mice that deep brain stimulation (DBS) of warm-sensitive neurones (WSNs) in the medial preoptic nucleus (MPN) produces tolerable hypothermia. In contrast to surface cooling-evoked hypothermia, DBS mice exhibit a torpor-like state without counteractive shivering. Like hypothermia evoked by chemogenetic activation of WSNs, DBS in free-moving mice elicits a rapid lowering of the core body temperature to 32-34 °C, which confers significant brain protection and motor function reservation. Mechanistically, activation of WSNs contributes to DBS-evoked hypothermia. Inhibition of WSNs prevents DBS-evoked hypothermia. Maintaining the core body temperature at normothermia during DBS abolishes DBS-mediated brain protection. Thus, the MPN is a DBS target to evoke tolerable therapeutic hypothermia for stroke treatment.

Peripheral Cold War: A Perspective Study of India and Pakistan Relations

India and Pakistan have long history of animosity which is evident in their bilateral relations. Defining the politics of security is an attempt to first develop the comprehensive definition of security in India Pakistan case and engender certain knowledge that how the threats are surfacing in other aspects of state affairs. The political, economic and social sectors have associated with security while strategic relations of India and United States to counter China are also very important aspects of Pakistan security proposition. This research paper seeks to have a latest oversight to observe how these changing relations might incriminate the strategic environment of South Asia and specifically in Pakistan India relations and how the strategic importance of these periphery’s lead them to cooperate with strong states to get the military and economic benefits. The conflict and strategic relations with the powerful states to maintain the balance of power, forms the peripheral cold war.

The Role of Butyrate in Paclitaxel and Morphine Induced Peripheral Hypersensitivity

Recent evidence implicates the gut microbiome in peripheral hypersensitivity in chronic opioid use models, and in chemotherapeutic‐induced hypersensitivity in mice. An important constituent of the commensal microbiome are bacterial species that ferment fibers through complex enzymatic pathways and produce short chain fatty acids, acetate, propionate, and butyrate. Butyrate is known to improve the integrity of the epithelium and enhance antimicrobial peptides thus preventing colonization by pathogenic bacteria, as well as having an established role in reducing inflammation through the inhibition of histone deacetylase. Based on this, we hypothesized that sodium butyrate can relieve peripheral hypersensitivity as well as reduce primary nociceptor hyperexcitability in experimental models of peripheral hypersensitivity. We used both paclitaxel (PAC) and morphine (MOR) animal models, which were organized into the following treatment groups: PAC (8mg/kg i.p q.d, total 4 injections) ± Sodium Butyrate (10mg/kg i.p b.i.d), MOR (escalating from 20mg/kg i.p b.i.d on day 1 to 80mg/kg i.p b.i.d on day 4) ± Sodium Butyrate (10mg/kg i.p b.i.d). PAC animals were assessed for peripheral cold hypersensitivity using an acetone evaporation assay. MOR‐treated animals were assayed using a hot‐plate test for opioid‐induced hyperalgesia. Primary nociceptors from L4‐S1 dorsal root ganglia were collected from these mice for whole‐cell patch clamp electrophysiological recordings to assess for enhanced excitability. PAC‐treated mice developed significant cold hypersensitivity 7 days post PAC, when compared with vehicle (average time engaging stimulated paw 2.0s vs 8.6s respectively). Butyrate ameliorated this paclitaxel‐induced hypersensitivity (average time reduced to 2.5s). Similarly, MOR induced thermal hypersensitivity (Hot‐plate latency of 14.8s vs 28.16s saline), which was also reversed by sodium butyrate (Hot‐plate latency of 24.84s). Whole‐cell patch clamp recordings revealed that PAC and MOR‐treated neurons fired a greater number of regenerating action potentials compared to respective controls (PAC: 4 vs 2; MOR: 3.8 vs 2.1), indicating enhanced excitability. In‐vivo butyrate treatment attenuated this enhanced excitability. Hysteresis plots of hyperexcitable cells revealed a depolarizing shift in the membrane potential of regenerating action potentials. Sodium butyrate treatment recovered this shift in membrane potential within the repetitive action potentials. Therefore, similar to the in‐vivo peripheral hypersensitivity, butyrate reversed enhanced excitability of PAC and MOR‐treated DRG neurons. These findings demonstrate that the short chain fatty acid, butyrate, a major metabolite of the gut microbiome plays an important role in preventing the development of drug‐induced peripheral hypersensitivity.

Dietary Bioflavanol Supplementation Does Not Improve Cold‐Induced Vasodilation in Healthy Adults

BackgroundCold‐induced vasodilation (CIVD) is a cyclical rise in blood flow to glabrous skin that occurs during cold exposure of the extremities. Nitric oxide (NO) is a proposed mediator of the CIVD response through active vasodilation and/or withdrawal of sympathetic vascular smooth muscle tone. Dietary flavanols augment bioavailable NO by upregulating NO synthase, the main source of NO in the vasculature, and by enhancing removal of reactive oxygen species that are known to breakdown NO. Although acute dietary flavanol intake has been shown to augment skin blood flow, at least in part through increased NO, no studies have examined the impact of flavanol intake on skin perfusion during cold exposure. Improving skin blood flow to the extremities may enhance manual dexterity and reduce the risk of peripheral cold injury, benefits pertinent to warfighters or other populations that spend time in cold environments.PurposeTo examine the effect of high‐dose flavanol supplementation on CIVD. We hypothesized that acute and 8‐day flavanol supplementation would augment CIVD during single‐digit cold water immersion.MethodsEleven healthy adults (24±6 yrs; 10M/1F) ingested cocoa‐based flavanols (FLV; 900 mg/d) or caffeine and theobromine‐matched placebo (PLA) for 8 days in a double‐blind, randomized, crossover design. On Days 1 and 8, CIVD was assessed 2 hours after treatment ingestion. Subjects immersed their middle finger in warm water (42°C) for 15 minutes before cold water immersion (4°C) for 30 min, during which nail bed and finger pad skin temperature were measured continuously.ResultsFlavanol ingestion had no effect (all P &gt; 0.05) on CIVD event frequency (Day 1, FLV: 3±2 vs. PLA: 3±2; Day 8, FLV: 3±2 vs. PLA: 3±1) or amplitude (Day 1, FLV: 4.3±1.7 vs. PLA: 4.9±2.6 °C; Day 8, FLV: 3.9±1.9 vs. PLA: 3.9±2.0 °C) in the finger pad following acute or 8‐day supplementation. Furthermore, average (Day 1, FLV: 9.2±1.5 vs. PLA: 9.2±1.8 °C; Day 8, FLV: 8.3±1.7 vs. PLA: 8.9±1.9 °C), maximum (Day 1, FLV: 11.9±3.0 vs. PLA: 11.2±2.3 °C; Day 8, FLV: 10.1±1.9 vs. PLA: 10.6±2.3 °C), and minimum (Day 1, FLV: 6.2±1.3 vs. PLA: 6.0±1.5 °C; Day 8, FLV: 5.8±1.8 vs. PLA: 6.2±1.7 °C) finger pad temperatures during cold water immersion were not different (all P &gt; 0.05) between treatments on Day 1 or Day 8 of supplementation. Similarly, no differences (all P &gt; 0.05) in CIVD parameters were observed in the nail bed following acute or 8‐day supplementation.ConclusionDietary ingestion of cocoa‐based flavanols does not improve the CIVD response to single‐digit cold water immersion.

Research on the Spatial Agglomeration Characteristics and Influencing Factors of Express Delivery Station Based on DNN.

In this paper, the POI data of 736 Cainiao stations in Nanjing is taken as the research sample. With the help of ArcGIS software, the standard deviation ellipse, spatial autocorrelation, average nearest neighbor, cold and hot spot analysis, nuclear density estimation, and other spatial analysis models are used to quantitatively characterize its business mode, spatial distribution characteristics, and equilibrium. Based on DNN, the spatial agglomeration characteristics and distribution directions of the Cainiao station in Nanjing were sorted out, the cold spots and hot spots of the spatial layout were identified, and the spatial differentiation rules and agglomeration patterns were revealed. Finally, the geographically weighted regression analysis model is used to analyze the influencing factors of the spatial agglomeration of the Cainiao station in Nanjing. The research found that Firstly, the proportion of Nanjing Cainiao station operating mode adopting the exclusive mode is 59.1%, the proportion adopting the concurrent operation mode is 33.7%, and the rest adopting the joint operation mode of cooperation with other logistics enterprises. Secondly, Nanjing Cainiao Station gathers in the central city area, forming a “central hot spot.” The urban fringe area does not form a “peripheral cold spot area,” and the whole presents a “1 + 4” five-core agglomeration model across the river. Thirdly, Regional GDP, population density, and the number of convenience stores/supermarkets are the main factors affecting the spatial agglomeration of the Cainiao station in Nanjing.

Role of TRPM8 in cold avoidance behaviors and brain activation during innocuous and nocuous cold stimuli

Transient receptor potential melastatin 8 (TRPM8) is a cold-sensing cation channel; however, its role in the transferal of information on peripheral cold sensation to the brain remains unclear. Therefore, we herein investigated cold avoidance behaviors and the neuronal activation of the hypothalamus and cerebral cortex in TRPM8 knockout (KO) mice to innocuous and nocuous cold stimuli. An innocuous cold stimulation at 15 °C decreased the duration of sleeping and increased that of rearing, climbing, and eating in WT mice, but it did not alter the duration of these behaviors in TRPM8 KO animals. The innocuous cold stimulation also increased the frequency of rearing, climbing, walking, and eating in WT mice, but it did not change that of these behaviors in TRPM8 KO animals. In contrast, a nocuous cold stimulation at 9 °C decreased the duration of sleeping and increased that of rearing and climbing in both WT and TRPM8 KO mice. The nocuous cold stimulation increased the frequency of rearing, climbing, and walking in WT and TRPM8 KO mice. Quantitative Fos immunohistochemistry showed that both innocuous and nocuous cold stimulations increased the number of Fos-positive neurons in temperature- and metabolism-associated hypothalamic regions in WT mice, but not in TRPM8 KO animals. The number of Fos-positive neurons was markedly increased in the primary motor and somatosensory cortices in WT and TRPM8 KO mice following the nocuous cold stimulation, but only increased in WT mice after the innocuous cold stimulation. Collectively, the present results indicate that TRPM8 plays a crucial role in activating autonomic hypothalamic neuronal circuits under innocuous and nocuous cold stimuli.

Peripheral Cold Sensation in Patients with Chronic Heart Failure

Peripheral Cold Sensation in Patients with Chronic Heart Failure

Surgical management of biatrial myxoma: case report

Heart tumours account for approximately 0.2 % of all tumours: of these, approximately 75 % of all primary heart tumours are benign and 50 % of them are myxomas. Further, myxomas make up 0.0017 % of the general population of patients with cardiovascular disease. Biatrial myxomas, i.e. tumours in both the left and right atria, can be in the form of a ‘butterfly’ or a ‘dumbbell’, and account for &lt; 1 % of all cardiac myxomas. Here we describe the successful surgical management of a rare case of a large biatrial myxoma and concomitant atrioventricular valve insufficiency. Briefly, 2D transthoracic echocardiography findings included an end-diastolic volume of 90 ml, an end-systolic volume of 40 ml and a left ventricular ejection fraction of 55 % (according to Simpson). The fibrous ring of the mitral valve measured 36 mm with a regurgitation degree of 2 while the fibrous ring of the tricuspid valve was 42 mm in size and the regurgitation grade was 3. Lesion size in the cavity of the left and right atrium were 73 × 38 mm and 80 × 42 mm, respectively. These neoplasia were surgically removed under peripheral cardiopulmonary bypass, hypothermia and cold cardioplegia (Custodiol). The myxomas were accessed through the right atrium, according to Giradon and were resected without defragmentation. Next, mitral valve plasty using a soft support ring and annuloplasty of the tricuspid valve were performed according to de Vega. The duration of artificial circulation was 150 minutes and that of myocardial ischaemia was 100 minutes. The patient was extubated 11 hours after surgery, spent 22 hours in the intensive care unit and was discharged on the 14th day after surgery. Surgical resection of biatrial myxomas before the development of serious irreversible or life-threatening complications can provide rapid symptomatic relief in congestive heart failure.Received 30 May 2021. Revised 22 August 2021. Accepted 23 August 2021.Funding: The study did not have sponsorship.Conflict of interest: Authors declare no conflict of interest.Contribution of the authors Literature review: T.A. Simonyan Drafting the article: T.A. Simonyan, I.M. Tsiskaridze Critical revision of the article: T.A. Simonyan, I.Yu. Farulova, E.A. Babajanyan Surgical treatment: I.I. Scopin, I.M. Tsiskaridze Final approval of the version to be published: T.A. Simonyan, I.I. Scopin, I.M. Tsiskaridze, I.Yu. Farulova, E.A. Babajanyan

Non-healing Ulcer on the Foot as Cellulitis: A Case Report

Introduction: Cellulitis is a serious bacterial skin infection. The skin is swollen and red, and it feels warm and uncomfortable to the touch. Cellulitis is most commonly found in the lower legs, although it can also appear on the face, arms, and other regions of the body. Bacteria enter the body through a crack or break in the skin. The infection can spread to your lymph nodes and bloodstream if left untreated, creating a major health danger. It is rarely passed down from one generation to the next.&#x0D; Clinical Findings: The patient presented with a non-healing ulcer for 30days, due to the insect bite 1month ago on right foot which is being developed into non-healing ulcer on right foot. A 20x6cm wound over the right foot was seen on a plain film radiograph of the knee. It consists of necrotic debris, underlying tendons exposed, peripheral cold and pigmentation. Foul smelling is not present. Based on the appearance of the spreading erythema and radiographically confirmed findings, cellulitis was diagnosed. &#x0D; Diagnostic Evaluation: Blood test- Hb-8.6gm%, MCV-91.1fl, MCH-30.9Pico-gm, MCHC-33.9%, Total RBC Count-2.78 millions/cu.mm, RDW-16.8%, HCT-25.3%, Total WBC Count-7400 cu.mm, Monocytes-04%, Granulocytes-75%, Lymphocytes-20%, Eosinophills-01%, Basophills-00%, Total Platelet Count-2.06lacs/cu.mm Peripheral Smear: RBCs-Normocytic hypochromic Platelets Adequate on smear, No Haemoparasite seen.&#x0D; Therapeutic Intervention: Inj. Ceftriaxone 1gm IV BD; Inj. PAN 40mg IV OD; Inj. Emset 4 mg IV TD; Inj. Neomal 100 ml IV TDS; Inj. MVI IV OD; Tab. Dolo 650 mg TDS; Tab Limcee 500 mg OD; Cap. Becosute OD; Protein powder; Tab Shelcal 500 mg OD; Tab. Duphalac 15 ml. &#x0D; Conclusion: My patient was admitted to Surgery Ward, A.V.B.R.H with a known case of Cellulitis and he had a complaint of non-healing-ulcer over the right limb. After getting appropriate treatment his condition was approved.

Constitutive Phosphorylation as a Key Regulator of TRPM8 Channel Function

In mammals, environmental cold sensing conducted by peripheral cold thermoreceptor neurons mostly depends on TRPM8, an ion channel that has evolved to become the main molecular cold transducer. This TRP channel is activated by cold, cooling compounds, such as menthol, voltage, and rises in osmolality. TRPM8 function is regulated by kinase activity that phosphorylates the channel under resting conditions. However, which specific residues, how this post-translational modification modulates TRPM8 activity, and its influence on cold sensing are still poorly understood. By mass spectrometry, we identified four serine residues within the N-terminus (S26, S29, S541, and S542) constitutively phosphorylated in the mouse ortholog. TRPM8 function was examined by Ca2+ imaging and patch-clamp recordings, revealing that treatment with staurosporine, a kinase inhibitor, augmented its cold- and menthol-evoked responses. S29A mutation is sufficient to increase TRPM8 activity, suggesting that phosphorylation of this residue is a central molecular determinant of this negative regulation. Biophysical and total internal reflection fluorescence-based analysis revealed a dual mechanism in the potentiated responses of unphosphorylated TRPM8: a shift in the voltage activation curve toward more negative potentials and an increase in the number of active channels at the plasma membrane. Importantly, basal kinase activity negatively modulates TRPM8 function at cold thermoreceptors from male and female mice, an observation accounted for by mathematical modeling. Overall, our findings suggest that cold temperature detection could be rapidly and reversibly fine-tuned by controlling the TRPM8 basal phosphorylation state, a mechanism that acts as a dynamic molecular brake of this thermo-TRP channel function in primary sensory neurons.SIGNIFICANCE STATEMENT Post-translational modifications are one of the main molecular mechanisms involved in adjusting the sensitivity of sensory ion channels to changing environmental conditions. Here we show, for the first time, that constitutive phosphorylation of the well-conserved serine 29 within the N-terminal domain negatively modulates TRPM8 channel activity, reducing its activation by agonists and decreasing the number of active channels at the plasma membrane. Basal phosphorylation of TRPM8 acts as a key regulator of its function as the main cold-transduction channel, significantly contributing to the net response of primary sensory neurons to temperature reductions. This reversible and dynamic modulatory mechanism opens new opportunities to regulate TRPM8 function in pathologic conditions where this thermo-TRP channel plays a critical role.

UCP1 and TRPM8 Expression in the Brown Fat Did Not Affect the Restriction of Menthol-Induced Hyperthermia by Estradiol in Ovariectomized Rats.

Estradiol (E2) modulates the central and peripheral thermoregulatory responses to cold. Menthol is an agonist of transient receptor potential melastatin type 8 (TRPM8), which is a peripheral cold receptor. E2 suppresses menthol-induced elevation of body temperature (Tb) in ovariectomized rats, but the mechanism is unknown. The aim of the present study is to investigate the effect of E2 on uncoupling protein 1 (UCP1), a thermogenic gene, and TRPM8 mRNA levels in ovariectomized rats applied menthol. A silastic tube was implanted in ovariectomized rats with and without E2 underneath the dorsal skin (E2(+) and E2(-) groups), and data loggers for Tb measurement into peritoneal cavity. After application of 10% L-menthol or vehicle to the skin of the whole trunk of rats, Tb was measured for 2 h. The interscapular brown adipose tissue (BAT) and spinal ganglia of cervical, thoracic, and lumbar parts were obtained for RT-qPCR assay. In the menthol application, Tb in the E2(+) group was lower than that in the E2(-) group. The UCP1 mRNA in the BAT, TRPM8 mRNA in the BAT and spinal ganglia in all areas did not differ between the E2(+) and E2(-) groups. In conclusion, the UCP1 and TRPM8 expression in the brown fat did not affect the restriction of the menthol-induced hyperthermia by estradiol in ovariectomized rats.

Kv4.3 Channel Dysfunction Contributes to Trigeminal Neuropathic Pain Manifested with Orofacial Cold Hypersensitivity in Rats

Trigeminal neuropathic pain is the most debilitating pain disorder but current treatments including opiates are not effective. A common symptom of trigeminal neuropathic pain is cold allodynia/hyperalgesia or cold hypersensitivity in orofacial area, a region where exposure to cooling temperatures are inevitable in daily life. Mechanisms underlying trigeminal neuropathic pain manifested with cold hypersensitivity are not fully understood. In this study, we investigated trigeminal neuropathic pain in male rats following infraorbital nerve chronic constrictive injury (ION-CCI). Assessed by the orofacial operant behavioral test, ION-CCI animals displayed orofacial cold hypersensitivity. The cold hypersensitivity was associated with the hyperexcitability of small-sized trigeminal ganglion (TG) neurons that innervated orofacial regions. Furthermore, ION-CCI resulted in a reduction of A-type voltage-gated K+currents (IA currents) in these TG neurons. We further showed that these small-sized TG neurons expressed Kv4.3 voltage-gated K+channels, and Kv4.3 expression in these cells was significantly downregulated following ION-CCI. Pharmacological inhibition of Kv4.3 channels with phrixotoxin-2 inhibited IA-currents in these TG neurons and induced orofacial cold hypersensitivity. On the other hand, pharmacological potentiation of Kv4.3 channels amplified IA currents in these TG neurons and alleviated orofacial cold hypersensitivity in ION-CCI rats. Collectively, Kv4.3 downregulation in nociceptive trigeminal afferent fibers may contribute to peripheral cold hypersensitivity following trigeminal nerve injury, and Kv4.3 activators may be clinically useful to alleviate trigeminal neuropathic pain.SIGNIFICANCE STATEMENTTrigeminal neuropathic pain, the most debilitating pain disorder, is often triggered and exacerbated by cooling temperatures. Here, we created infraorbital nerve chronic constrictive injury (ION-CCI) in rats, an animal model of trigeminal neuropathic pain to show that dysfunction of Kv4.3 voltage-gated K+channels in nociceptive-like trigeminal ganglion (TG) neurons underlies the trigeminal neuropathic pain manifested with cold hypersensitivity in orofacial regions. Furthermore, we demonstrate that pharmacological potentiation of Kv4.3 channels can alleviate orofacial cold hypersensitivity in ION-CCI rats. Our results may have clinical implications in trigeminal neuropathic pain in human patients, and Kv4.3 channels may be an effective therapeutic target for this devastating pain disorder.

The Effect of Cold on the Trigeminal Reflexes

Objective: Effects of muscle cooling on the spinal stretch and monosynaptic reflexes have been studied to describe the properties of Group I and II afferents of muscle spindles. Masseter muscle differs from extremity muscles in structural and numerical features of the muscle spindles. The aim of this study was to examine the muscle spindle afferent features of masseter muscle by applying cold to understand the role of Group I and II afferents in reflexes of masseter. Patients and Methods: We included 12 healthy subjects (7 females and 5 males) in the study. Masseter inhibitory reflex (MIR), jaw tendon reflex (JTR), trigeminal motor evoked potential (MEP), and trigeminal somatosensory evoked potential (SEP) were studied before and after cold application. Left masseter muscle was cooled down to 18°C. We compared the data obtained before and after cold application. Results: After cold application, the mean total duration of MIR was shortened and it was absent in four subjects. The mean amplitude of JTR was higher after cold application (P = 0.018) without any significant change in latency. The mean latency of MEP was delayed without any change in amplitude (P = 0.003). There was no significant difference in SEPs. Conclusions: Changes observed in MIR, JTR, and MEP could not be ascribed to any specific type of muscle spindle afferents. Delayed mean reflex latencies were attributed to the effect of cold on nerve conduction. Summation of peripheral cold and pain receptor features, spindle afferents, and cortical mechanisms might have caused cold-associated changes.

The effect of inter-pulse delay on the spectral emission and expansion dynamics of plasma in dual-pulse fiber-optic laser-induced breakdown spectroscopy

The role of inter-pulse delay on plasma dynamics and spectral emission in dual-pulse fiber-delivery plasma has been investigated using fast imaging, optical emission spectroscopy, and laser shadowgraphy. The detection on the return spectrum in dual-pulse fiber-optic laser-induced breakdown spectroscopy showed that the self-reversal and self-absorption were reduced as the inter-pulse delay increased from 50 to 1000 ns. Using scanning electron microscopy, the ablation depth showed a changing trend of increasing first and then decreasing, and a maximum of ∼2.8 μm was achieved at 250-ns inter-pulse delay. Experimental results confirmed that the improvement was due to the reduction of the plasma thickness from 1.161 mm to 0.964 mm, and the calculation of electronic excitation temperature along the photon collection path showed that the excited atom densities became more spread, which both contributed to the self-absorption reduction. At a long inter-pulse delay, the plasma trailing edge would gradually separate from the target surface with a little further expansion distance of the leading edge, resulting in the reduction of plasma thickness after a time delay of several hundred nanoseconds. The heating of the peripheral cold particles by the second-generation plasma in the later expansion led to the rapid reduction of the peak temperature. Also, the early expansion trajectory of the second-generation plasma was tracked by laser shadowgraphy, which started to appear at an inter-pulse delay of 100 ns, and the average expansion velocity reached its maximum of ∼3.8 km/s.

Effects of a Standardized Oligomerized-Polyphenol from Litchi chinensis Fruit Extract and Mixed Plant Extract Supplementation on Peripheral Circulation and Cold Sensitivity.

Certain individuals tend to suffer from a cold sensation-particularly in the lower extremities-despite most people not suffering from the same sensation. In Japan, this phenomenon is called "hie-sho" and reduces quality of life for several people, particularly women. A previous study has shown that a standardized oligomerized-polyphenol from Litchi chinensis fruit extract (OPLFE) reportedly causes a significant increase in body surface temperature. The present study aimed to investigate whether supplementation with OPLFE affected peripheral circulation and cold sensitivity. This randomized, double-blind, placebo-controlled trial was performed including 25 participants (age, 45.0±10.4 y; 3 males and 22 females) who were assigned to consume OPLFE, mixed plant extract with OPLFE, or placebo capsules for 14 d. Participants were instructed to relax for 60 min in a temperature-controlled room prior to obtaining measurements. Changes in skin temperature and peripheral blood flow of the middle finger were assessed immediately before and 1, 5, 10, 20, and 30 min after immersion in cold water (10ºC). Participants' height, weight, skin temperature, and blood flow in peripheral tissue were measured; furthermore, their "hie-sho" was measured using the Visual Analog Scale (VAS). Skin temperature and blood flow in peripheral tissue increased in the OPLFE and mixed plant extract with OPLFE groups on day 14 compared with those on day 1. In addition, cold sensitivity in these two groups significantly improved between day 1 and day 14. These findings suggest that OPLFE improves "hie-sho" by increasing peripheral blood flow and skin temperature.