We thank Evelyn Lailey and Claudia Silva for their excellent technical assistance; the Canadian Foundation for Innovation for providing key infrastructure. Dr K. D. Patel and Dr C. Power are both SCH727965 order Canada Research Chairs. KDP is an Alberta Heritage Foundation for Medical Research Scientist and CP is an AHFMR Senior Scholar. Dr V.E.L. Stubbs is supported by fellowships from the Alzheimer Society of Canada, the CIHR Institute of Aging and the

CIHR Strategic Training Program. This research was supported by grants from the Heart and Stroke Foundation and the CIHR. None. “
“Epigenetic deregulation of genes encoded on the X chromosome as reported for CD40L in lupus could explain the female predominance of autoimmune

diseases. We compared CD40L expression on CD4+ T cells from primary Sjögren’s syndrome (pSS) women and healthy controls and investigated DNA methylation patterns of the promoter and enhancer regions of CD40L. The expression of CD40L on activated CD4+ T cells was higher in patients with pSS than controls after phorbolmyristate acetate and ionomycin activation (P = 0.02). CD40L mRNA level in CD4+ T cells did not differ between patients with pSS and controls and was similar in both groups in cultures treated with the demethylating agent 5-azacytidine C. Pyrosequencing analysis revealed no Obeticholic Acid manufacturer significant differences in methylation profiles between patients and controls. Inducible membrane-bound CD40L on CD4+ T cells is increased in patients with pSS but was not related to epigenetic deregulation by demethylation patterns of the regulatory regions of CD40L. Primary Sjögren’s syndrome (pSS) and systemic lupus erythematosus Methane monooxygenase (SLE) are two autoimmune diseases that share numerous pathogenic features and

the same strong predominance among women. The reason for the female preponderance is not yet understood but may be related to the X chromosome. In fact, one X chromosome in women is silenced by epigenetic mechanisms, so epigenetic deregulation could contribute to the female predisposition to autoimmunity via overexpression of some X-chromosome-located genes, and thus X-inactivation escape [1]. Numerous genes (e.g. Toll-like receptor 7 and CD40L) involved in adaptive and/or in innate immunity are located on the X chromosome. In a recent study of women with systemic sclerosis (SSc), 40% of patients versus 8% of controls showed skewed X chromosome inactivation (odds ratio 9.3, 95% confidence interval [95% CI] 4.3–20.6) [1]. In two other studies of SLE [2] and SSc [3], the promoter and downstream enhancer regions of CD40 ligand (CD40L) were hypomethylated. In pSS, overexpression of the soluble form of CD40L was reported [4, 5], but data are lacking on membrane-bound CD40L expression on CD4+ T cells. CD40L is a co-stimulation molecule of 260 amino acids located on Xq26.3–27.1. The gene consists of five exons and five introns.

Mesenchymal stromal cell (MSC) -mediated immunosuppression is non-cognate dependent and non-antigen-specific. The effector mechanisms prevalently involve soluble factors that are used by other immunomodulatory populations that are also recruited by the MSC. Mesenchymal stromal cells expand and activate regulatory T cells and interfere with the maturation and function of antigen-presenting

cells (APC). The interaction between MSC and haemopoietic stroma is fundamental because MSC depend on the presence of inflammatory molecules produced by monocytes/macrophages to become immunosuppressive. The inflammatory profile to which MSC are exposed determines their immunomodulatory properties, because only in the presence of cytokines like selleck screening library interferon-γ (IFN-γ) or tumour necrosis factor-α (TNF-α) do MSC become immunosuppressive (‘licensing’). Alternative stimulations polarize MSC towards a pro-inflammatory activity. More study of the physiological significance of the immunomodulatory activity is needed to better clarify their key role among the effectors of innate tolerance. AZD5363 cost It is not surprising that

MSC have generated enormous interest for therapeutic applications. Their properties have been extensively and successfully tested in animal models and in the clinical setting on a variety of autoimmune and alloimmune diseases but the modalities of the therapeutic efficacy remain to be elucidated. Although the existence of a population of MSC has long been recognized in many adult tissues, it was only recently that these cells received centre-stage attention. The characterization of MSC within the bone marrow, initially described in the 1960s

by Friedenstein et al.,[1, 2] paved the way to a number of studies that identified in this population a large proportion C225 of self-renewing progenitors capable of differentiating into adipocytes, osteoblasts and chondrocytes.[3-5] Since then, MSC with similar phenotypes and properties have been isolated from a number of other sources, including cord blood, adipose tissue, muscle and liver.[6-8] These findings led to the use of the acronym MSC to indicate mesenchymal stem cells, irrespective of their source, differentiation stage and function. In contrast to haemopoietic stem cells, the absence of an in vivo assay for quantifying their stemness/multipotency has hindered the identification of markers that can convincingly distinguish primitive stem cells from progenitors and the even less defined fibroblasts. Human MSC are reported as expressing CD105, CD73, CD90, CD44, CD71 and Stro-1, as well as the adhesion molecules CD106 [vascular cell adhesion molecule 1 (VCAM-1)], CD166, CD54 [intercellular adhesion molecule 1 (ICAM-1)] and CD29, in the absence of any haemopoietic markers.[9-12] The identity of murine MSC has progressed recently.

Guinea-pigs that were administered wild-type S. flexneri 2a and treated with opium post 4 days starvation developed fatal enteric infections (Formal et al., 1958). Because of the fatal effects at a relatively early stage of infection, this model was not ideal for the purpose of screening vaccine candidates. Although the rabbit shigellosis model was sensitive (Rabbani et al., 1995), its suitability for measuring the protection is not known. Rhesus monkeys are the only animals in which typical bacillary dysentery can be induced by oral infection with shigellae without starvation and/or pretreatment

with antibiotics (Takeuchi et al., 1968; Rout et al., 1975; Collins et al., 2008). However, the use of this animal selleck kinase inhibitor is a major constraint due to many reasons. Recently, a new guinea-pig model has been described that represents typical bacillary dysentery and acute rectocolitis after rectal inoculation (Shim et al., 2007). In this model, the catheter does not reach the

proximal colon, which is the specific site of Shigella colonization. In addition, backflow of inoculum cannot be prevented while removing the catheter. Considering the difficulties OTX015 mouse in the several animal models and methods, luminal inoculation in guinea-pigs is more reliable as this model allows Shigella to be retained in the proximal colon. Recently, Jeong et al. (2010) successfully developed a model of intragastric infection in 1–3-day-old piglets that induced symptoms and characteristic gut lesions similar to those of humans. The need for specialized isolators, environmentally controlled accommodation, competent animal handlers and labor-intensive systems are some of the issues that make this model unfavorable. The guinea-pig luminal model described in this study is ideal for studying Roflumilast bacillary dysentery in vivo as it covers several features such as the appropriate infection site, immune responsiveness and protective immunity. Thus, this model is ideal for the generation of preclinical information of Shigella vaccines before human volunteer studies. This model cannot entirely replace primate or human studies, but it can be used to generate preclinical

information that should significantly reduce the number of studies in primates as well as in humans. This work was supported by funds from the Indian Council of Medical Research, New Delhi, India, and the Japan Initiative for Global Research Network on Infectious Diseases (J-GRID), Ministry of Education, Culture, Sports, Science and Technology of Japan. S.B., Research Associate, is a recipient of J-GRID fellowship. The authors thank Mr Suhasit Ranjan Ghosh for technical assistance, Mr Prasanta Karmakar for graphical presentation and Mr Subhadip Dan for editorial assistance. “
“Citation Rose JA, Rabenold JJ, Parast MM, Milstone DS, Abrahams VM, Riley JK. Peptidoglycan induces necrosis and regulates cytokine production in murine trophoblast stem cells.

A free flap transfer combined Metformin with an autologous vein graft can cover large tissue defects and simultaneously improve distal perfusion even in patients with arterial occlusive disease. We are presenting a case of bypass-free radial forearm flap used to cover a foot defect in an old diabetic

patient with peripheral arterial disease. The flap perfusion deteriorated significantly during the early postoperative period. The patient was brought back to the operating room with acute thrombosis of the popliteal-radial venous graft and the arterial pedicle of the flap. The flap was salvaged by thrombectomy and creation of an additional arteriovenous fistula at the distal arterial pedicle. The procedure improved the flap perfusion and decreased the high internal resistance that was noticed in the flap when trying to flush the radial artery during the revision surgery and was evident by continuous wave -Doppler sonography. The successful salvage of the flap in the presented case and the convenient long-term follow up suggest that this technique may be safe and helpful as a last effort to salvage a bypass-free flap with a suspected high internal resistance. © 2013 Wiley Periodicals, Inc. Microsurgery 33:391–395, Protein Tyrosine Kinase inhibitor 2013. “
“Although ischemia-reperfusion (I/R) strongly influences muscle flap survival in reconstructive

surgery, there is limited knowledge about its relation to hemorheological parameters and oxidative stress markers in flaps. In the present study we investigated these changes during I/R of latissimus dorsi muscle (LDM) flaps in beagle dogs. In four animals LDM flaps were prepared bilaterally. The right side served as control, while the left side’s vascular pedicle was clamped for 60 minutes, and a 60-minute reperfusion was allowed afterward. Blood samples (0.5 ml each) were taken from the pedicle’s vein bilaterally before and after the ischemia,

and at the 5th, 15th, PLEKHM2 30th, 45th, and 60th minutes of the reperfusion, for hematological and erythrocyte aggregation tests. In muscle biopsies, taken before and after I/R, histological investigations and tests for measuring gluthation-peroxidase (GSH-PX) activity, glutathione (GSH) and carbonyl concentrations, and thiobarbituric acid reactive substances (TBARS) content were carried out. In I/R side leukocyte count increased during the reperfusion with a peak at the 30th minute. Hematocrit continuously increased from the 15th minute. In the first 5 minutes of the reperfusion, erythrocyte aggregation increased, than tented to be normalized. In muscle homogenates GSH-PX activity did not change markedly, GSH content slightly decreased, carbonyl and TBARS content increased during reperfusion. A 1-hour ischemia and reperfusion of LDM flaps caused local changes of leukocyte distribution and erythrocyte aggregation, supposedly due to the metabolic and inflammatory reactions.

We therefore isolated F5 T cells and determined their rate of death in vitro. T cells from control F5 donors ((F5 Rag1−/−×C57Bl6/J.CD45.1)F1, IL-7R+ F5 hereon) underwent check details progressive apoptosis over several days that was prevented by addition of IL-7 (Fig. 1A). In the absence of continued IL-7Rα expression in vivo, IL-7R– F5 T cells disappear relatively fast, with a half life of ∼14 d (Supporting Information Fig. 1), a phenotype that implies their reduced homeostatic fitness. Interestingly, upon culture in vitro, IL-7R– F5 T cells underwent apoptosis far more rapidly than controls, particularly at early time points (Fig. 1A). As expected,

in the absence of IL-7Rα expression, death of IL-7R– F5 T cells was not prevented by addition of IL-7 (Fig. 1A). We next examined the effect of non-limiting IL-7 in vivo on T-cell fitness. Control F5 T

cells were transferred into T-cell-deficient Rag1−/− hosts in which there is consequently no T-cell competition for IL-7. www.selleckchem.com/products/MK-1775.html Although F5 T cells proliferate in response to lymphopenia in Rag1−/− hosts, they retain a naïve phenotype 25, 26. After 7–14 d, survival of transferred cells was compared with T cells from intact F5 donors. Remarkably, F5 T cells recovered from Rag1−/− hosts survived in vitro in the complete absence of any survival or growth factors for many days (Fig. 1B), and exogenous IL-7 had little additional effect on their survival. T-cell survival was Bcl2 dependent, since addition of specific inhibitor ABT-737 caused death of all cells by 24 h (data not shown). Although naïve T cells proliferate in lymphopenic hosts, persistence of F5 T cells in vitro was a function of survival and not cell division, as F5 T cells did not continue to divide in vitro, even in the presence of exogenous IL-7 (Supporting Information Fig. 2A). While not further enhancing survival, IL-7 did maintain the increased cell size observed in F5 T cells

transferred to Rag1−/− hosts, suggesting that the trophic properties Liothyronine Sodium of IL-7 are more short lived and do require persistent IL-7 signalling (Supporting Information Fig. 2B) and also confirmed that IL-7 signalling had ceased in IL-7 free cultures. In Rag1−/− hosts, there is a lack of T-cell competition for other factors important for CD8+ T-cell survival, such as DCs expressing self-peptide–MHC (spMHC) and IL-15, which could also influence the fitness of F5 T cells. Additionally, IL-7Rα is also a component of the heterodimeric thymic stromal lymphopoietin (TSLP) receptor, that has also been implicated in maintenance of naïve CD4+ T cells 27, 28, and loss of signalling through this receptor could also be contributing to death of IL-7R– F5 T cells. Therefore we directly addressed the role of IL-7 in enhancing T-cell fitness by transferring the same cells to IL-7-deficient Rag1−/− mice.

c.). Mast cell numbers typically average about 9–10/mm2 of intestinal mucosa in uninfected hamsters (18), and the values in Figure 3 for naïve control animals (Group 1) concur. Likewise Group 3 hamsters (primary abbreviated infection), which had been treated to remove worms on day 35, recovered almost completely by day 73, showing mast cell

densities much like those of naïve animals on both days 73 and 94 of the experiment. In marked contrast hamsters that had experienced the uninterrupted primary infection (Group 2) had markedly elevated levels of mast cells, approximately five times more cells per mm2 of mucosal tissue on both days 73 and 94 p.i. Group 4 animals (secondary infection only) did not have elevated mast cell densities Dabrafenib mw on day 10 p.i., but by 31 days p.i. the numbers had increased approximately three fold. Unexpectedly, 10 days p.c. mast cell numbers in immunized, challenged hamsters (Group 5, primary + secondary infections) were much like those of the naïve animals and then rose only

slowly, although significantly, over the course of the remainder of the experiment (regression of mast cells/mm2 of mucosal tissue on days after challenge, confined to Group 5; Rp = 0·50, n = 20, β = 0·29 ± 0·118, t = 2·43, P = 0·026). Goblet cell numbers in naive hamsters usually average about 50–70/mm2 (18), and the values in Figure 4 for naive hamsters (Group 1) and those from which worms had been removed selleck Selleckchem Erastin (Group 3, primary abbreviated infection), fall comfortably within the normal range. In hamsters with an uninterrupted primary infection (Group 2), goblet cell numbers were two fold higher on day

73 p.i. and over three fold higher on day 94 p.i., and in Group 4, given only the second infection, they were about half as high on day 10 p.i. and twice as high on day 31 p.i. In contrast, hamsters in Group 5 (primary + secondary infection), goblet cell numbers on day 10 were within the naïve control range, but then climbed steeply to peak on day 24 more than four fold higher before dropping somewhat by day 31 p.c. The curve thus generated was best described by the quadratic equation y = −193·9 + 29·72x−0·6×2 (where y = goblet cells/mm2 and x = days after challenge); R2 = 52·2%, F2,17 = 11·36, P = 0·0007). Eosinophil counts averaged below 32 cells/mm2 in naive animals (Group 1), and in animals, which had been treated to remove worms (Group 3, primary abbreviated infection) the values were about twice higher, but averaging below 66 cells/mm2 (Figure 5). In contrast in hamsters with the uninterrupted primary infection (Group 2) on days 73 and 94 p.i., the eosinophil counts were 12·8 and 9·7-fold higher, respectively, relative to the appropriate naïve control group.

These cells stimulate T helper type 1 (Th1) helper cells that in turn elicit the production of cytotoxic T lymphocytes (CTL) [22]. These cytotoxic effector cells attack infected cells, resulting in resolution of the infection [23]. However, little is known about how to modulate these immune responses. Prophylactic vaccination. 

Vaccination Autophagy inhibitor with VLPs gives rise to virus-neutralizing antibodies in serum. Vaccination by intramuscular injection of L1 VLPs has been shown to be highly immunogenic and well tolerated in Phase I trials [24–27]. Three randomized placebo-controlled Phase II trials with, respectively, a monovalent HPV16 vaccine, a bivalent HV16/18 vaccine and a quadrivalent HPV6/11/16/18 vaccine candidate have consistently demonstrated almost complete protection against persistent infection with the targeted HPV types [28–32]. Moreover, these trials confirmed Opaganib nmr the safety of the vaccines and showed strong immunoresponses that were several orders of magnitude higher than those observed after natural infections. Two pharmaceutical companies [Merck Sharp & Dohme (MSD) and GlaxoSmithKline (GSK)] have completed large multi-centre Phase III vaccine trials

in all continents except Africa [33–35]. In addition, the National Cancer Institute (United States) is conducting a population-based trial in Costa Rica using the bivalent vaccine [36]. These Phase III trials demonstrated that vaccines protect against histologically confirmed high-grade cervical intraepithelial neoplasia (CIN) and adenocarcinoma in situ (AIS) associated with the targeted HPV types under the condition that subjects were not infected with one or more vaccine types at baseline [33–35]. Both vaccine formulations have a good safety profile. Selleckchem Enzalutamide Neither has noted any therapeutic effect, as women who test positive for HPV DNA prior to vaccination show no protection against disease end-points associated with that type. Modest cross-protection to closely related high-risk types HPV

31, 33, 45 was found with bivalent vaccine [Cervarix(R)][37] and also to some extent with the quadrivalent vaccine [Gardasil(R)][38,39]. Therapeutic HPV vaccines.  Development of cervical precursors, their maintenance and progression to invasive cancer requires the continued intracellular expression of the viral oncoproteins E6 and E7 [40,41]. Therefore, therapeutic vaccines have been directed towards stimulating T cell responses against these viral early oncogenes. The approaches include administration of peptide antigens or recombinant proteins, plasmid DNA vaccines, viral vector vaccines and administration of E7-pulsed dendritic cells, but despite being variably immunogenic have not shown an impact upon invasive cancer but appear to induce some degree of clearance of cancer precursors or anogenital warts [23,42–44].

One week after the last immunization, mice were killed, blood was taken and, following perfusion, intestinal samples were collected using the perfusion-extraction (PERFEXT) technique.20 Ovalbumin-specific IgG and IgA titres were determined by ELISA. Doxorubicin Ninety-six-well plates (Greiner Bioscience, Frickenhausen, Germany) were coated with OVA (20 μg/ml)

and blocked with PBS/BSA. Serially diluted serum and intestinal samples were added followed by goat anti-mouse horseradish peroxidase-conjugated IgA or IgG (SouthernBiotech, Birmingham, AL). Plates were developed with o-phenylenediamine dihydrochloride, stopped with 0·1 m H2SO4 and absorbance was read at 490 nm. Titres of IgG and IgA were determined from the sample dilution giving an optical density value above 0·4. Data were statistically analysed in Prism (graphpad software) using the Student’s t-test, in which *P < 0·05, **P < 0·01 and ***P < 0·001. Although systemic immune compartments and skin-draining LN of CD47−/− mice have been extensively studied, the GALT has not been carefully characterized. We

therefore enumerated cells in the GALT of CD47−/− mice and revealed a 50% reduction of total cell numbers in MLN, LP and PP, compared with those in WT mice (Table 1). In contrast, the number of cells in skin-draining LN and spleen was not significantly different between WT and CD47−/− mice (Table 1). Although immunohistochemical analysis showed normal localization of T and B cells in MLN and PP of CD47−/− mice high throughput screening compounds (see supplementary material, Fig. S1a), and both CD47−/− and WT CD4+ T cells in PP and MLN were found to express similar levels of CD44 and CD62L (data not shown), the frequency of CD4+ T cells in MLN and PP of CD47−/− mice was significantly reduced compared with that in WT mice (Fig. S1b). In contrast, the frequency of Foxp3+ CD4+ T cells in PP, but not in MLN, was significantly increased in CD47−/− compared with WT mice (Fig. S1c). Impaired DC migration from the skin and subset-specific Sclareol alterations in splenic DC at steady state have previously been

reported in CD47−/− mice13,14 therefore, we next assessed populations of antigen-presenting cells in the GALT of these mice. As the total number of cells in the GALT of CD47−/− mice was reduced by 50%, frequency rather than total number of cells within cell populations was determined. Flow cytometric analysis showed a significant reduction in the frequency of CD11c+ MHC-II+ conventional DC (cDC) in MLN, but not in LP or PP, of CD47−/− mice (Fig. 1a). In contrast, no significant change in the frequency of CD172a+ CD11clow MHC-IIlow SSClow cells was detected (Fig. 1b). Further phenotypic characterization was therefore focused on cDC and identified two populations of cDC in MLN (see supplementary material, Fig. S2a).

However, in B cells, receptor internalization occurs within 15 min [9, 42]. The differential kinetics in actin oxidation between the cell types could control the differences in actin reorganization following

activation. Interestingly, in B cells, SHP-1 maximal oxidation occurred at 5 min and was similar to CD8+ T cells [8]. Previous work has shown that recruitment of SHP-1 to CD22 is necessary to downregulate BCR signals [43]. Docking of SHP-1 to CD22 could explain the delay in oxidation, ensuring that SHP-1 activity is decreased when it is recruited to the plasma membrane to allow full signal through the BCR. Furthermore, we are the first to document that PTEN is oxidized following B-cell activation. Like SHP-1, cysteine

oxidation of PTEN and ZVADFMK its subsequent inactivation could be delayed allowing the opposing kinase, PI3K, to dock at CD19 [44]. Interestingly, we could not detect sulfenic acid formation in CD45 following B-cell activation. It is possible that CD45 could be in a disulfide bond with glutathione, sulfenamide, sulfinic, or sulfonic acid species, which may account for our inability to detect sulfenic acid. Together, our results demonstrate that B cells exhibit this website a unique cysteine oxidation profile following activation compared to other cell types and it is tightly regulated to facilitate proper signal transduction and activation. In this study, we demonstrate that the reversible oxidation of cysteine is a mechanism by which ROI modifies proteins to promote B-cell activation and proliferation. The goal of autoimmune therapies selleckchem and vaccination is to dampen or enhance the immune response, respectively. By identifying proteins in signaling pathways that are regulated by oxidation, it may be possible to design targeted therapeutics to modulate B-cell

responses. Spleens were removed from 6- to 8-week-old C57BL/6 mice after cervical dislocation. After teasing apart the spleen on a wire mesh screen, red blood cells were osmotically lysed using ACK Lysis Buffer (Lonza). Splenocytes were resuspended in complete media composed of RPMI 1640 supplemented with 10% fetal calf serum (FCS, HyClone), L-glutamine (HyClone), penicillin-streptomycin (Cellgro), nonessential amino acids (GIBCO), and 2-mercaptoethanol (GIBCO). All animal studies were approved by the Institutional Animal Care and Use Committee (IACUC) of the Wake Forest University School of Medicine. B cells were isolated from spleens of C57BL/6 mice using Miltenyi Biotec CD43 negative selection magnetic bead separation according to the manufacturer’s protocol. Purity was routinely greater than 96% B220+ cells as determined by acquisition on FACSCalibur instrument. For all stimulations, with the exception of the calcium flux experiments, purified cells were pretreated for 1 h at 37°C with complete media alone (vehicle) or media containing dimedone (Sigma-Aldrich).

In the process of surveying the CIVD literature, one major caveat is the lack of consensus

in definition and protocols, making it very difficult to compare results across studies. As seen in Figure 1, many parameters are used to quantify CIVD, and studies may report improved CIVD simply from a change in a single parameter, with other parameters either not significant or not reported. At its most basic, Selleck Talazoparib no standardized definition for a rise in digit skin temperature that constitutes a CIVD event exists, with some studies depending on a deflection of skin temperature from a baseline [1,16,49], through to temperature increases ranging from 0.5°C to 4.0°C as a CIVD threshold [28,36]. Similar variability exists in quantifying what is actually meant by an improved thermal response across studies, which can consist of factors, such as more numerous CIVD events, higher mean or minimum digit temperatures, or more rapid onset times for CIVD [15]. To overcome these methodological differences, we carefully read the methodology of each

study and determined to report the parameters that are most common for CIVD research like onset time and mean, minimum, maximum finger/toe temperatures. Another example of methodological variability across studies is the use of either skin blood flow or skin temperature, each of which may be measured with different types of sensors at different sites, as generally interchangeable methods for measuring CIVD. Tamoxifen chemical structure While the transposition of blood flow and skin temperature may appear intuitive, little direct evidence exists. Shitzer et al. [69] modeled and experimentally validated the relationship between blood perfusion in the fingertips during cold exposure with finger skin temperature, whereas Daanen [14] reported that skin perfusion preceded the temperature response by 112 ± 72 seconds with a cross-correlation coefficient of 0.76 ± 0.14. Figure 3 illustrates the many different responses possible from cold exposure, further making quantification of CIVD difficult. For research in this field to advance, it seems critical that basic standardized definitions

and protocols be adopted Axenfeld syndrome to maximize the integration of research. O’Brien’s [59] study on the reproducibility of CIVD may provide a starting point for standardization of ambient and individual factors; a number of individual factors were standardized in a study on the reproducibility of CIVD, including circadian rhythm, pre-test nutrition, posture, site of sensor placement, and pre-immersion in warm water to normalize vasodilation. Other experimental factors may need to be controlled, especially depth of digit or limb immersion [68], along with ambient or core temperatures due to the strong relationship between body temperature and CIVD response [19,25], and the demonstration that facial protection improved finger temperature and thermal comfort during whole-body cold exposure [60].