Preliminary study of finger temperature recovery in patients with diabetes mellitus following cold stimulation.

OBJECTIVE: To explore the difference in temperature recovery following cold stimulation between participants with and without diabetes mellitus (DM). MATERIALS AND METHODS: The participants without (control group; n = 25) and with (DM group; n = 26) DM were subjected to local cold stimulation (10º C for 90 s). The thermal images of their hands were continuously captured using a thermal camera within 7 min following cold stimulation, and the highest temperature of each fingertip was calculated. According to the temperature values at different timepoints, the temperature recovery curves were drawn, and the baseline temperature (T-base), initial temperature after cooling (T0), temperature decline amplitude (T-range), and area under the temperature recovery curve > T0 (S) were calculated. Finally, symmetry differences between the two groups were analysed. RESULTS: No statistical differences in the T-base, T0, and T-range were observed between the DM and control groups. After drawing the rewarming curve according to the temperature of the fingertips of the patients following cold stimulation, the S in the DM group was significantly lower than that in the control group (p < 0.05). Furthermore, the asymmetry of the base temperature of the hand was observed in the DM group. CONCLUSIONS: Following cold stimulation, the patients with DM exhibited a different rewarming pattern than those without DM. Thus, cold stimulation tests under infrared thermography may contribute to the early screening of diabetic peripheral neuropathy in future.

Cold stimulation causes oxidative stress, inflammatory response and apoptosis in broiler heart via regulating Nrf2/HO-1 and NF-κB pathway.

To investigate the effect of cold stimulation on heart, 300 1-day-old female broilers were divided into control (CON) and two cold stimulation (CS3 and CS9) groups. Birds in CON group were reared in normal ambient temperature during day 1-43; while birds in CS3 and CS9 groups were reared at 3 °C and 9 °C below CON group for 5 h at 1-day intervals from day 15 to day 35, respectively. Heart tissues were collected at day 22, 29, 36, and 43 to determine the indexes related to oxidative stress, inflammation and apoptosis. The H&E staining displayed that inflammatory cell infiltration and myocardial fiber break were obviously observed in CS9 group, and cardiac pathological score in CS9 group was higher than CON and CS3 groups (P < 0.05) at day 22, 36, and 43. Overall, compared to CON group, the concentrations of MDA and H2O2 were elevated, the activities of SOD, CAT, GPx, and T-AOC were reduced, and mRNA expression of CAT, GPx, SOD, nuclear factor-erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) was downregulated in CS9 group at each time-point (P < 0.05). Compared to CON group, mRNA expression of NF-κBp65, COX-2, iNOS, PTGEs, TNF-α, and IL-1ß, and mRNA and protein expression of Bax, Bak, Cyt-c, caspase-3, and caspase-9 were increased, while Bcl-2 and Bcl-2/Bax ratio were decreased in CS9 group (P < 0.05) at the most detected time-points. There were no significant differences in the levels of indexes associated with oxidative stress, Nrf2/HO-1 antioxidant system, inflammation, and apoptosis between CON and CS3 groups at the most detected time-points (P > 0.05). Therefore, this study suggests that severe cold stimulation at 9 °C below normal rearing temperature induces cardiomyocyte inflammation and apoptosis by regulating Nrf2/HO-1 pathway-related oxidative stress in broilers, and mild cold stimulation of CS3 group can improve the adaptability of hearts to cold environment.

Effects of alternating heat and cold stimulation at different cooling rates using a wearable thermo device on shoulder muscle stiffness: a cross-over study.

BACKGROUND: A small, wearable thermo device that uses Peltier elements for programmed heat and cold stimulation has been developed recently and is expected to be applied in conventional contrast bath therapy. This study was aimed to examine improvements in trapezius muscle hardness and subjective symptoms resulting from alternating heat and cold stimulation, with different rates of cooling. METHODS: This cross-over study included four conditions. Twenty healthy young male individuals (age, 22.3 ± 4.5 years) participated in this study. These four interventions targeted the unilateral trapezius muscle of the dominant arm after a 15-min typing task. Specifically, heat and cold stimulations were applied at different ratios (the heating/cooling rate of 3:1, 3:2, and 3:3) or not applied. Each intervention was separated by at least one week. Skin temperature at the stimulation area was recorded using a data logger. Outcome measures included muscle hardness (measured using a portable tester) and subjective symptoms (muscle stiffness and fatigue). Each item was assessed at three time points: baseline, after typing, and after the intervention. RESULTS: Two-way analysis of variance with repeated measures found an interaction effect for muscle hardness between four conditions (3:1, 3:2, 3:3, and no) and three time points (p < 0.05). Only in the 3:1 condition were the post-intervention values lower than those after typing (p < 0.01). There was also an interaction effect for subjective muscle stiffness (p < 0.05); the values after the intervention in the 3:1 condition were lower than those after intervention in the no stimulation condition (p < 0.01). There was no significant relationship between changes in muscle hardness and changes in subjective symptoms in the 3:1 condition. CONCLUSIONS: Our results demonstrate that alternating heat and cold stimulations with a different cooling rate could affect the degree of improvement in muscle hardness and subjective symptoms. In particular, the 3:1 condition has the possibility to improved muscle hardness within the condition and subjective muscle stiffness between conditions. TRIAL REGISTRATION: UMIN000040620. Registered 1 June 2020, https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000046359.

Letter to Editor Regarding the Article Entitled "Impact of Sensory Stimulation on Pharyngo-esophageal Swallowing Biomechanics in Adults with Dysphagia: A High-Resolution Manometry Study".

In accordance with the abstract as well as results of the article entitled "Impact of Sensory Stimulation on Pharyngo-esophageal Swallowing Biomechanics in Adults with Dysphagia: A High-Resolution Manometry Study" published in Dysphagia journal, it has been found the cold bolus significantly reduced extent of upper esophageal sphincter opening during swallowing as measured by Integrated Relaxation Pressure from 9.34 to 5.17 mmHg. But, contrarily, it is written in paragraph four of the discussion that "interestingly, only the cold bolus significantly increased the extent of upper esophageal sphincter opening in this study based on the Integrated Relaxation Pressure metric" which considerably contrasts with the result of the study.

Procyanidin B2 alleviates liver injury caused by cold stimulation through Sonic hedgehog signalling and autophagy.

Procyanidin B2 (PB2), a naturally occurring flavonoid abundant in a wide range of fruits, has been shown to exert antioxidant, anti-inflammatory and anticancer properties. However, the role of PB2 in the prevention of cold stimulation (CS)-induced liver injury. The present study was undertaken to determine the effects of PB2 on liver injury induced by cold stimulation and its potential molecular mechanisms. The present study results showed that treatment with PB2 significantly reduced CS-induced liver injury by alleviating histopathological changes and serum levels of alanine transaminase and aspartate transaminase. Moreover, treatment with PB2 inhibited secretion of inflammatory cytokines and oxidative stress in cold-stimulated mice. PB2 reduced cold stimulation-induced inflammation by inhibiting TLR4/NF-κB and Txnip/NLRP3 signalling. Treatment with PB2 reduced oxidative stress by activating Nrf-2/Keap1, AMPK/GSK3ß signalling pathways and autophagy. Furthermore, simultaneous application of Shh pathway inhibitor cyclopamine proved that PB2 targets the Hh pathway. More importantly, co-treatment with PB2 and cyclopamine showed better efficacy than monotherapy. In conclusion, our findings provide new evidence that PB2 has protective potential against CS-induced liver injury, which might be closely linked to the inhibition of Shh signalling pathway.

Cold urticaria - What we know and what we do not know.

Cold urticaria (ColdU) is a common form of chronic inducible urticaria characterized by the development of wheals, angioedema or both in response to cold exposure. Recent research and guideline updates have advanced our understanding and management of ColdU. Today, its pathophysiology is thought to involve the cold-induced formation of autoallergens and IgE to these autoallergens, which provoke a release of proinflammatory mediators from skin mast cells. The classification of ColdU includes typical and atypical subtypes. We know that cold-induced wheals usually develop on rewarming and resolve within an hour and that anaphylaxis can occur. The diagnosis relies on the patient's history and cold stimulation testing. Additional diagnostic work-up, including a search for underlying infections, should only be done if indicated by the patient's history. The management of ColdU includes cold avoidance, the regular use of nonsedating antihistamines and the off-label use of omalizumab. However, many questions regarding ColdU remain unanswered. Here, we review what is known about ColdU, and we present important unanswered questions on the epidemiology, underlying pathomechanisms, clinical heterogeneity and treatment outcomes. Our aim is to guide future efforts that will close these knowledge gaps and advance the management of ColdU.

Reduced brown adipose tissue-associated skin temperature following cold stimulation in children and adolescents with type 1 diabetes.

BACKGROUND: Brown adipose tissue (BAT) is essential to maintain body temperature. Its ability to convert chemical energy in glucose and free fatty acids to heat is conferred by a unique protein, UCP-1. BAT activity is greatest in children and adolescents, declining through adulthood. Blood glucose concentrations outside the normal nondiabetic range are common in type 1 diabetes and hyperglycaemia leads to insulin resistance in muscle and white adipose tissue, but whether this applies to BAT, is not known. METHOD: To investigate the effect of type 1 diabetes on BAT activity, we measured the supraclavicular temperature of 20 children with type 1 diabetes and compared them to 20 age-matched controls, using infrared thermography. RESULTS: The diabetes group had lower stimulated supraclavicular temperatures (diabetes group: 35.03 (34.76-35.30)°C; control group: 35.42 (35.16-35.69)°C; p = 0.037) and a reduced response in relative temperature following cold stimulation, after adjusting for BMI (diabetes group: 0.11 (0.03-0.18)°C; control group: 0.22 (0.15-0.29)°C; p = 0.034). In the diabetes group, there was no association between glycaemic measures and supraclavicular temperatures, but the method of insulin delivery may significantly affect the change in supraclavicular temperature with stimulation (injections: 0.01 (-0.07-0.09)°C; pump: 0.15 (0.04-0.26)°C; p = 0.028). CONCLUSIONS: While further work is needed to better understand the glucose-insulin-BAT relationship, one possible explanation for the reduced supraclavicular temperature is that exogenous, unlike endogenous, insulin, is not suppressed by the activity of the sympathetic nervous system, preventing lipolysis-driven activation of BAT.

Modulation of the cutaneous and cortical silent period in response to local menthol application.

In this study, we investigated the effects of menthol application on the cortical and cutaneous silent period (CSP/cutSP). Both the cutSP and CSP were assessed while participants (n = 11, young adults) exerted a light contraction with the right thumb. In the 1st block of trials, SPs were measured after the application of a Neutral gel (Aloe Vera) to the dorsal aspect of the hand. In the 2nd block, the same measures were repeated following a Menthol gel (4%) application. Subjective ratings of cooling sensations were obtained for each block. The Neutral gel was consistently perceived as slightly cool by participants, wheres the Menthol gel elicited sensations from cool to very cold. Paired t-tests showed no difference in the cutSP duration between the two conditions, whereas a significant increase in the CSP was detected with the Menthol condition. No correlation was found between changes in the CSP and those of the cutSP. These results highlight the difference between the cutSP and the CSP, as inhibitory phenomena, and point to a cortical contribution to the soothing effects associated with topical menthol applications.

Effect of cold oral stimulation on orthostatic hypotension in multiple system atrophy: a case study.

[Purpose] The aim of this case study is to reconsider the method for preventing orthostatic hypotension in multiple system atrophy. [Participant and Methods] The case was that of a 70-year-old female with multiple system atrophy who experienced frequent falls and orthostatic hypotension. An orthostatic test was performed, and the effect of cold oral stimulation before standing was compared with no stimulation. Outcome measures were blood pressure, heart rate variability and autonomic variables. [Results] In the control test, blood pressure decreased from 150/72 mmHg in the supine position to 98/58 mmHg in the standing position. Heart rate increased from 71 bpm to 82 bpm, high frequency declined from 16.48 msec2 to 14.07 msec2, and low/high frequency increased from 2.56 to 5.13. Cold stimulation in the standing position induced changes in blood pressure from 168/82 mmHg to 104/72 mmHg, heart rate from 73 bpm to 83 bpm, high frequency from 61.29 msec2 to 24.56 msec2, and low/high frequency from 1.45 to 6.33 msec2. [Conclusion] Standing after cold stimulation affected autonomic variables, but did not affect the heart rate or blood pressure, possibly because of damaged peripheral blood vessels. Further research is required to demonstrate the effect of cold stimulation on orthostatic hypotension.

Pre-cold acclimation improves the immune function of trachea and resistance to cold stress in broilers.

Acclimation can alleviate the damage caused by adverse environmental factors. To investigate the effects of cold stimulation on immunity in tracheal of broilers, 360 one-day-old chicks were raised at normal temperatures during 1-7 days. From 8 day, G1 (control) continued to be raised at normal temperatures, whereas G2 and G3 (treatment groups) were cold-stimulated at 3°C and 12°C below the temperature of G1, respectively. At 42 day, all the groups were subjected to a 24-hr acute cold stress, designated as S1, S2, and S3. Tracheal tissues were collected to detect gene levels of immunoglobulins, antimicrobial peptides, Hsps, and cytokines, and oxidative stress-related indicators at 14 day, 42 day, and 43 day, and protein levels of Hsps and proinflammatory cytokines as well as morphology changes at 42 day and 43 day. The results showed that, compared with 42G1, tracheal structure of 42G2 was basically intact, and gene levels of immunoglobulins and antimicrobial peptides increased (p < 0.05), whereas tracheal structure of 42G3 was destroyed, with decreased levels of immunoglobulins ( p < 0.05), and increased levels of Hsps and proinflammatory cytokines ( p < 0.05). At 43 day, tracheal damage was visible and gene levels of immunoglobulins and antimicrobial peptides decreased in S1 ( p < 0.05). Tracheal structure was relatively intact and gene levels of antimicrobial peptides increased in S2 ( p < 0.05). Compared with S1 and S3, immune-related gene levels in S2 were higher, and Hsps and proinflammatory cytokines levels were lower. The results demonstrate that cold stimulation of lower 3°C from 8 to 42 day led to cold acclimation, which improved immunity of tracheal mucosa and resistance to cold stress in broilers.

The effects of skin cold stimulation on quadriceps muscle activity during walking in older adults.

Purpose: Given that walking speed declines with ageing and decreasing walking speed restricts activities of daily living (ADL), it is important for the old to maintain walking speed in order to prevent affecting ADL. Although skin cold stimulation (SCS) facilitates instantaneous muscle activity, which occurs during walking, the effects of SCS on muscle activity during walking remain unclear. Thus, the present study aimed to investigate the effect of SCS during walking in older adults.Methods: Seventeen community-dwelling healthy older adults (73 ± 6 years old) participated in this study. Walking speed at a comfortable pace and the electromyographic (EMG) activity of the vastus lateralis (VL) and biceps femoris (BF) were measured. SCS, which maintains the skin temperature at 25 °C, was applied to the front of the thigh during the procedures. Walking speed, root mean square EMG (rmsEMG) and mean power frequency (MPF) were compared under SCS and control conditions.Results: SCS significantly increased the walking speed (p < 0.01) and the rmsEMG of the vastus lateralis (p = 0.032). No change in the rmsEMG of the BF was observed, and SCS had no effect on MPF of both the VL and BF. Furthermore, a significant relationship was observed between these changes (r = 0.619, p = 0.042).Conclusion: SCS increased the EMG activity of the VL while increasing walking speed. Our results suggest that SCS is an effective strategy that can be included in daily life in order to improve walking ability of older adults.

Transcriptional Response of Subcutaneous White Adipose Tissue to Acute Cold Exposure in Mice.

Beige adipose tissue has been considered to have potential applications in combating obesity and its related metabolic diseases. However, the mechanisms of acute cold-stimulated beige formation still remain largely unknown. Here, transcriptional analysis of acute cold-stimulated (4 °C for 4 h) subcutaneous white adipose tissue (sWAT) was conducted to determine the molecular signatures that might be involved in beige formation. Histological analysis confirmed the appearance of beige adipocytes in acute cold-treated sWAT. The RNA-sequencing data revealed that 714 genes were differentially expressed (p-value < 0.05 and fold change > 2), in which 221 genes were upregulated and 493 genes were downregulated. Gene Ontology (GO) analyses showed that the upregulated genes were enriched in the GO terms related to lipid metabolic process, fatty acid metabolic process, lipid oxidation, fatty acid oxidation, etc. In contrast, downregulated genes were assigned the GO terms of regulation of immune response, regulation of response to stimulus, defense response, etc. The expressions of some browning candidate genes were validated in cold-treated sWAT and 3T3-L1 cell browning differentiation. In summary, our results illustrated the transcriptional response of sWAT to acute cold exposure and identified the genes, including Acad11, Cyp2e1, Plin5, and Pdk2, involved in beige adipocyte formation in mice.

Induction of copper and iron in acute cold-stimulated brown adipose tissues.

Copper (Cu) and iron (Fe), the enzymatic cofactors required for various biochemical processes, have been recently linked to lipid metabolism. Adipose tissues play central roles in energy balance and whether copper and iron homeostasis is involved in cold-stimulated energy metabolism remains unknown. In this study, the Cu and Fe contents in interscapular brown adipose tissue (BAT) and subcutaneous white adipose tissue (subWAT) from mice at different developmental stages were measured. Our results revealed the dynamic changes of Cu and Fe levels during development, suggesting their potential roles in energy homeostasis. Furthermore, the significantly increased Cu and Fe contents and the markedly up-regulated CTR1 and ATP7A expression in acute cold-stimulated BAT indicates the involvement of both Cu and Fe in the BAT-mediated thermogenesis. Comparably, no change in the Cu and Fe levels in cold-stimulated subWAT was observed at the indicated time points, suggesting that both metals are not involved in the cellular process of cold-induced WAT browning. Taken together, these data demonstrate the induction of Cu and Fe in acute cold-stimulated BAT activation, thus providing the direct evidences of the involvement of copper and iron homeostasis in BAT-mediated non-shivering energy metabolism.

A Pilot Study Investigating Clinical Responses and Biological Pathways of Azelastine/Fluticasone in Nonallergic Vasomotor Rhinitis before and after Cold Dry Air Provocation.

BACKGROUND: Nonallergic vasomotor rhinitis (NAVMR) has been considered a diagnosis by exclusion due to unknown mechanisms or lack of diagnostic biomarkers. METHODS: To determine clinical responses and biological pathways in NAVMR subjects challenged to cold dry air (CDA) in an environmental exposure chamber (EEC) pre- and posttreatment with azelastine/fluticasone (AzeFlu), 30 NAVMR subjects, prescreened for CDA-induced symptoms (approx. 14°C, <15% relative humidity, ×1 h) were randomized to treatment with AzeFlu (n = 20) or placebo (n = 10) for 2 weeks. Total nasal symptoms scores, minimum cross-sectional area, cough, and conjunctival redness were recorded at visit 1 (pretreatment) and visit 2 (posttreatment) before, during, and after CDA challenge. At both visits, nasal lavage fluid (NLF) and nasal scrapings (NS) were collected pre- and post-CDA challenge. Substance P, neurokinin-A, and calcitonin gene-related peptide concentrations in NLF were analyzed pre- and postchallenge at each visit. Their relationship with CDA-induced symptoms was determined by statistical analysis. MicroRNA sequencing from NS determined differentially expressed miRNA between the treatment groups post-CDA challenge at each visit. RESULTS: The minimum cross-sectional area (p < 0.05), cough count (p < 0.05), and substance P (p < 0.01) improved posttreatment with AzeFlu versus placebo. Gene targets for differentially expressed miRNAs at visit 1 were enriched for biological pathways regulating epithelial ciliogenesis and cell integrity that were modified in the AzeFlu-treated group versus placebo posttreatment. CONCLUSIONS: This study demonstrated the feasibility of an EEC model to investigate CDA-induced clinical responses and pathobiology in NAVMR subjects pre- and posttreatment with AzeFlu. NAVMR disease mechanisms for other nonallergic triggers can be investigated similarly.

Potential effects of pleasant and cold stimuli on nausea and vomiting induced by disgusting tastes.

Several pharmacological agents have disgusting tastes that are perceived strongly in the back of the mouth and may trigger nausea and vomiting (NV), resulting in poor adherence to medication schedules and negative impacts on clinical outcomes. Pleasant stimuli and cold temperature lessen the disgusting stimuli, lowering NV through different mechanisms. A pleasant stimulus can mask an unpleasant one, presumably through lateral inhibitory connections in the local neuronal circuit. Similarly, temperature deeply influences taste perception because the response to bitter as well as to salty and sour has been found to assume a reversed U-shaped form, being reduced by cooling to 18°C and enhanced by warming to 30-37°C. This Review describes the mechanisms by which pleasant and cold stimuli may mask emetogenic disgusting stimuli and identifies the potential clinical applications of cooling for inhibiting objectionable drug-related gustatory reactions. © 2015 Wiley Periodicals, Inc.

Segmentation and characterization of interscapular brown adipose tissue in rats by multi-parametric magnetic resonance imaging.

OBJECTIVE: The aim was to auto-segment and characterize brown adipose, white adipose and muscle tissues in rats by multi-parametric magnetic resonance imaging with validation by histology and UCP1. MATERIALS AND METHODS: Male Wistar rats were randomized into two groups for thermoneutral (n = 8) and cold exposure (n = 8) interventions, and quantitative MRI was performed longitudinally at 7 and 11 weeks. Prior to imaging, rats were maintained at either thermoneutral body temperature (36 ± 0.5 °C), or short term cold exposure (26 ± 0.5 °C). Neural network based automatic segmentation was performed on multi-parametric images including fat fraction, T2 and T2* maps. Isolated tissues were subjected to histology and UCP1 analysis. RESULTS: Multi-parametric approach showed precise delineation of the interscapular brown adipose tissue (iBAT), white adipose tissue (WAT) and muscle regions. Neural network based segmentation results were compared with manually drawn regions of interest, and showed 96.6 and 97.1% accuracy for WAT and BAT respectively. Longitudinal assessment of the iBAT volumes showed a reduction at 11 weeks of age compared to 7 weeks. The cold exposed group showed increased iBAT volume compared to thermoneutral group at both 7 and 11 weeks. Histology and UCP1 expression analysis supported our imaging results. CONCLUSION: Multi-parametric MR based neural network auto-segmentation provides accurate separation of BAT, WAT and muscle tissues in the interscapular region. The cold exposure improves the classification and quantification of heterogeneous BAT.

Wide-scale quantitative phosphoproteomic analysis reveals that cold treatment of T cells closely mimics soluble antibody stimulation.

The activation of T lymphocytes through antigen-mediated T cell receptor (TCR) clustering is vital in regulating the adaptive immune response. Although T cell receptor signaling has been extensively studied, the fundamental mechanisms for signal initiation are not fully understood. Reduced temperatures have initiated some of the hallmarks of TCR signaling, such as increased phosphorylation and activation on ERK and calcium release from the endoplasmic reticulum, as well as coalesced the T cell membrane microdomains. The precise mechanism of the TCR signaling initiation due to temperature change remains obscure. One critical question is whether the signaling initiated by the cold treatment of T cells differs from the signaling initiated by the cross-linking of the T cell receptor. To address this uncertainty, we performed a wide-scale, quantitative mass-spectrometry-based phosphoproteomic analysis on T cells stimulated either by temperature shifts or through the cross-linking of the TCR. Careful statistical comparisons between the two stimulations revealed a striking level of identity among the subset of 339 sites that changed significantly with both stimulations. This study demonstrates for the first time, in unprecedented detail, that T cell cold treatment was sufficient to initiate signaling patterns that were nearly identical to those of soluble antibody stimulation, shedding new light on the mechanism of activation of these critically important immune cells.

Intermittent mild cold stimulation alleviates cold stress-induced pulmonary fibrosis by inhibiting the TGF-ß1/Smad signaling pathway in broilers.

To investigate the potential protective effect of intermittent cold stimulation on lung tissues of broilers exposed to acute cold stress (ACS). A total of 384 one-day-old broilers were assigned to 4 experimental groups with 6 replicates of 16 birds each: control (CON) and ACS groups were reared at normal feeding temperature from d 1 to 42; cold treatment groups (CS3+ACS and CS9+ACS) were reared, respectively, at 3°C or 9°C for 5 h on alternate days below the CON group from d 15 to 35. Animals in CS3+ACS, CS9+ACS, and ACS groups were exposed at 10°C for 24 h on d 43. Subsequently, lung tissues were collected to perform histopathological examination and measurement of relevant indexes. The results showed that lung tissues in CS9+ACS and ACS groups exhibited increased inflammatory cell infiltrates and collagen deposition compared to the CON group, while this pathological phenomenon was less pronounced in the CS3+ACS group. Compared to CON group, H2O2 and MDA contents were increased, and the activities of antioxidant enzymes (CAT, SOD, GPx, T-AOC) were reduced in CS9+ACS and ACS group (P < 0.05); mRNA and protein levels of inhibitor of NF-κB, Smad7, matrix metallopeptidase (MMP)-2, MMP9, and antioxidant-related genes were downregulated, whereas mRNA and protein levels of genes related to NF-κB/NLRP3 pathway-regulated inflammation and TGF-ß1/Smad pathway-regulated fibrosis were upregulated in cold-stressed broilers (P < 0.05). mRNA levels of heme oxygenase-1, NAD(P)H quinone oxidoreductase-1, and MMP9 were increased in CS3+ACS group (P < 0.05). Moreover, the expression of most antioxidant-related genes was increased, and that of inflammation- and fibrosis-related genes was reduced in CS3+ACS group (P < 0.05). Therefore, cold stress caused oxidative stress and inflammation, leading to pulmonary fibrosis in broilers, whereas intermittent mild cold stimulation at 3°C below normal rearing temperature alleviated fibrosis by inhibiting the TGF-ß1/Smad pathway modulated by the Nrf2/HO-1 and NF-κB/NLRP3 signaling pathway. This study suggests that intermittent mild cold stimulation can be a potential strategy to reduce ACS-induced lung damage in broilers.

Intermittent mild cold acclimation ameliorates intestinal inflammation and immune dysfunction in acute cold-stressed broilers by regulating the TLR4/MyD88/NF-κB pathway.

To investigate the potential protective effect of prior cold stimulation on broiler intestine induced by acute cold stress (ACS). A total of 384 one-day-old broilers were divided into control (CON), ACS, cold stimulation Ⅰ (CS3+ACS), and cold stimulation Ⅱ (CS9+ACS) groups. Broilers in CON and ACS groups were reared normally, and birds in CS3+ACS and CS9+ACS groups were reared at 3℃ and 9℃ below CON group for 5 h, respectively, on alternate days from d 15 to 35. Broilers in ACS, CS3+ACS, and CS9+ACS groups were subjected to 10℃ for 24 h on d 43. Eventually, small intestine tissues were collected for histopathological observation and indexes detection. The results showed that intestinal tissues in all ACS-broilers exhibited inflammatory cell infiltrates, microvilli disruption, reduced villus length in jejunum and increased crypt depth in jejunum and ileum. Whereas these phenomena were relatively light in CS3+ACS group. Compared to CON group, mRNA expression of the TLR4/MyD88/NF-κB pathway-related genes (TLR4, MyD88, NF-κBp65, COX-2, iNOS, PTGEs, TNF-α), Th1/Th17-derived cytokines (IL-1ß, IL-2, IL-8, IL-12, IFN-γ, IL-17), and HSPs (HSP40, HSP60, HSP70, HSP90) was upregulated (P < 0.05), and that of Th2-deviated cytokines (IL-4, IL-6, IL-10, IL-13) and IκBα was downregulated (P < 0.05) in small intestine in almost all ACS-broilers. Compared to ACS group, mRNA expression of most of the TLR4/MyD88/NF-κB pathway-related genes, Th1/Th17-derived cytokines, and HSPs was downregulated and that of Th2-derived cytokines was upregulated in CS3+ACS group (P < 0.05). Protein expression levels of TLR4, MyD88, p-p65/p65, p-IκBα/IκBα, IKK, TNF-α, IL-1ß, IL-10, and HSPs were similar to their mRNA expression. The concentration of sIgA and activities of CAT, SOD, and GSH-px were decreased and MDA and H2O2 were increased in ACS and CS9+ACS groups compared to CON group (P < 0.05). Therefore, cold stress caused oxidative stress and inflammation, leading to gut immune dysfunction; while mild cold stimulation at 3℃ below normal rearing temperature alleviated cold stress-induced intestinal injure and dysfunction by modulating the TLR4/MyD88/NF-κB pathway in broilers.

Study on the effect of human sympathetic nerve cold stimulation to relieve driving fatigue based on order recurrence plot.

Driving fatigue is very likely to cause traffic accidents, seriously threatening the lives and properties of drivers. Therefore, accurate detection and effective mitigation of driving fatigue are crucial for ensuring the personal safety of drivers. This study proposes a method to relieve driving fatigue by properly reducing the temperature to stimulate the human sympathetic nerve. The method uses the intelligent cooling and blowing device on the car seat cushion to achieve cold stimulation of the sympathetic nerve of the driver by reducing the temperature of the driver's hip, back and neck, so as to increase the excitement of the sympathetic nerve, keep the driver alert and achieve the purpose of fighting driving fatigue. In view of the fact that the traditional fatigue detection method is easily affected by environmental factors and individual differences, this study uses the order recurrence plot (ORP) method to detect driving fatigue based on electroencephalogram (EEG) signals. The results show that ORP textures drawn by EEG signals of the two driving conditions (normal driving condition and sensory cold stimulation driving condition) are significantly different, and the quantization parameters determinism (DET) and average diagonal line length (DLL) values are significantly different. Cold stimulation of the subjects' hips, back and neck to alleviate driving fatigue was the best when the temperature was 21 °C. In addition, compared with the traditional methods of fatigue relief, the sensory cold stimulation method proposed in this study does not easily to produce tolerance and has no damage to the body.