Human Experimental Models Research Articles

Cannabidiol reverses attentional bias to cigarette cues in a human experimental model of tobacco withdrawal.

Background and AimsCannabidiol (CBD), a non‐intoxicating cannabinoid found in cannabis, may be a promising novel smoking cessation treatment due to its anxiolytic properties, minimal side effects and research showing that it may modify drug cue salience. We used an experimental medicine approach with dependent cigarette smokers to investigate if (1) overnight nicotine abstinence, compared with satiety, will produce greater attentional bias (AB), higher pleasantness ratings of cigarette‐related stimuli and increased craving and withdrawal; and (2) CBD in comparison to placebo, would attenuate AB, pleasantness of cigarette‐related stimuli, craving and withdrawal and not produce any side effects.DesignRandomized, double‐blind cross‐over study with a fixed satiated session followed by two overnight abstinent sessions.SettingUK laboratory.ParticipantsThirty non‐treatment‐seeking, dependent cigarette smokers recruited from the community.Intervention and comparator800 mg oral CBD, or matched placebo (PBO) in a counterbalanced orderMeasurementsAB to pictorial tobacco cues was recorded using a visual probe task and an explicit rating task. Withdrawal, craving, side effects, heart rate and blood pressure were assessed repeatedly.FindingsWhen participants received PBO, tobacco abstinence increased AB (P = 0.001, d = 0.789) compared with satiety. However, CBD reversed this effect, such that automatic AB was directed away from cigarette cues (P = 0.007, d = 0.704) and no longer differed from satiety (P = 0.82). Compared with PBO, CBD also reduced explicit pleasantness of cigarette images (P = 0.011; d = 0.514). Craving (Bayes factor = 7.08) and withdrawal (Bayes factor = 6.95) were unaffected by CBD, but greater in abstinence compared with satiety. Systolic blood pressure decreased under CBD during abstinence.ConclusionsA single 800‐mg oral dose of cannabidiol reduced the salience and pleasantness of cigarette cues, compared with placebo, after overnight cigarette abstinence in dependent smokers. Cannabidiol did not influence tobacco craving or withdrawal or any subjectively rated side effects.

Creation of Experimental Human Nose Model With Lyophilized and Decellularized Bovine Cartilage Xenograft.

The nose anatomy is a functional and aesthetically important organ because of its three-dimensional structure, visible location in the face region, and its connection with the respiratory tract. Aesthetic and reconstructive nasal surgery requires correction of deformations in cartilage and bone structures as well as preservation of the natural connections between all subunits. The minimal mistake made can result in functional or aesthetically bad results. In this study, the authors aimed to create an experimental nose model that help aesthetic and reconstructive nose surgery operations.

Pro-Inflammatory Th1 and Th17 Cells Are Suppressed During Human Experimental Endotoxemia Whereas Anti-Inflammatory IL-10 Producing T-Cells Are Unaffected.

Sepsis is one of the leading causes of the deaths in hospitals. During sepsis, patients are exposed to endotoxemia, which may contribute to the dysregulation of the immune system frequently observed in sepsis. This dysregulation leads to impaired pro-inflammatory responses and may increase the risk for secondary infections in sepsis. The experimental human endotoxemia model is widely used as a model system to study the acute effects of endotoxemia. Under physiological circumstances, the immune system is tightly regulated. Effector T-cells exert pro-inflammatory function and are restrained by regulatory T-cells (Tregs), which modulate pro-inflammatory effector responses. Endotoxemia may induce inadequate Treg activity or render effector T-cells dysfunctional. It was the aim of the study to investigate effector T-cell and Treg responses in an experimental human endotoxemia model. In a cross-over designed placebo-controlled study, 20 healthy male volunteers received an intravenous injection of either lipopolysaccharide (LPS) (0.8 ng/kg body weight) or a placebo (saline 0.9%). CD3+ T-cells, CD4+ T-cells, CD8+ T-cells, and intracellular cytokine profiles were measured with flow cytometry at baseline and at repeated points after LPS/placebo injection. Complete blood cell counts were obtained with an automated hematology analyzer and cytokines were quantified by ELISA. Circulating neutrophils were significantly increased 2 h after LPS injection (p < 0.001) while absolute number of CD3+ T-cells, CD4+ T-cells, and CD8+ T-cells decreased (p < 0.001). Effector T-helper-cells (THs) showed a significant-but transient-decrease of pro-inflammatory IFNγ, interleukin (IL)-2, TNFα, and IL-17A production after LPS injection (p < 0.001). In contrast, the frequency of Treg and the capacity to produce IL-10 were unchanged (p = 0.21). Effector THs fail to produce pro-inflammatory Th1-/Th17-associated cytokines after LPS challenge. In contrast, IL-10 production by Treg is not affected. Thus, endotoxemia-induced suppression of pro-inflammatory THs might be considered as a contributing factor to immunoparalysis in sepsis.

Using Multiplex and Single-Molecule Bead-Based Technologies to Assess Inflammatory Changes Following Skeletal Muscle Injury

Abstract Inflammation is a key component in the healing and recovery of injured skeletal muscle. Despite the known importance of inflammation, its response has not been consistently demonstrated in human experimental model systems. One plausible explanation for the inconsistent outcomes in human skeletal muscle injury models may be caused by the analytical methods selected. Traditional methods (i.e. ELISA, Western Blotting, etc.) used to measure serum inflammatory/muscle damage biomarkers lack the bandwidth and/or sensitivity to measure multiple biomarkers or femtomolar changes. The purpose of this investigation was to describe a comprehensive method for the measurement of serum inflammatory biomarkers in a human skeletal muscle injury model. To accomplish this objective, we combined 384-well, bead-based multiplex assays (MILLIPLEX® and Luminex® FM3D) with a novel, bead-based single molecule detection technology (MilliporeSigma SMCxPro). The samples used in the present investigation were collected from an IRB-approved study and consented subjects (in accordance with the latest Declaration of Helsinki). Subjects completed a muscle damage exercise session and blood samples were collected at 0, 24, 48, and 72-h after muscle damage. We observed a significant change in inflammation post-damage using both detection technologies. Combining multiplexing with the precise, single molecule counting detection method allowed for a better clarification and description of the inflammatory response in otherwise undetectable sample concentrations. More research is needed to demonstrate repeatability for this assay approach as well as validation for other inflammatory states (i.e. chronic inflammatory disease).

Type I interferons in tuberculosis: Foe and occasionally friend.

Tuberculosis remains one of the leading causes of mortality worldwide, and, despite its clinical significance, there are still significant gaps in our understanding of pathogenic and protective mechanisms triggered by Mycobacterium tuberculosis infection. Type I interferons (IFN) regulate a broad family of genes that either stimulate or inhibit immune function, having both host-protective and detrimental effects, and exhibit well-characterized antiviral activity. Transcriptional studies have uncovered a potential deleterious role for type I IFN in active tuberculosis. Since then, additional studies in human tuberculosis and experimental mouse models of M. tuberculosis infection support the concept that type I IFN promotes both bacterial expansion and disease pathogenesis. More recently, studies in a different setting have suggested a putative protective role for type I IFN. In this study, we discuss the mechanistic and contextual factors that determine the detrimental versus beneficial outcomes of type I IFN induction during M. tuberculosis infection, from human disease to experimental mouse models of tuberculosis.

T271. Cannabidiol Reverses Attentional Bias to Cigarette Cues in a Human Experimental Model of Tobacco Withdrawal

T271. Cannabidiol Reverses Attentional Bias to Cigarette Cues in a Human Experimental Model of Tobacco Withdrawal

Identification of homocysteine-suppressive mitochondrial ETC complex genes and tissue expression profile – Novel hypothesis establishment

Hyperhomocysteinemia (HHcy) is an independent risk factor for cardiovascular disease (CVD) which has been implicated in matochondrial (Mt) function impairment. In this study, we characterized Hcy metabolism in mouse tissues by using LC-ESI-MS/MS analysis, established tissue expression profiles for 84 nuclear-encoded Mt electron transport chain complex (nMt-ETC-Com) genes in 20 human and 19 mouse tissues by database mining, and modeled the effect of HHcy on Mt-ETC function. Hcy levels were high in mouse kidney/lung/spleen/liver (24–14 nmol/g tissue) but low in brain/heart (~5 nmol/g). S-adenosylhomocysteine (SAH) levels were high in the liver/kidney (59–33 nmol/g), moderate in lung/heart/brain (7–4 nmol/g) and low in spleen (1 nmol/g). S-adenosylmethionine (SAM) was comparable in all tissues (42–18 nmol/g). SAM/SAH ratio was as high as 25.6 in the spleen but much lower in the heart/lung/brain/kidney/liver (7–0.6). The nMt-ETC-Com genes were highly expressed in muscle/pituitary gland/heart/BM in humans and in lymph node/heart/pancreas/brain in mice. We identified 15 Hcy-suppressive nMt-ETC-Com genes whose mRNA levels were negatively correlated with tissue Hcy levels, including 11 complex-I, one complex-IV and two complex-V genes. Among the 11 Hcy-suppressive complex-I genes, 4 are complex-I core subunits. Based on the pattern of tissue expression of these genes, we classified tissues into three tiers (high/mid/low-Hcy responsive), and defined heart/eye/pancreas/brain/kidney/liver/testis/embryonic tissues as tier 1 (high-Hcy responsive) tissues in both human and mice. Furthermore, through extensive literature mining, we found that most of the Hcy-suppressive nMt-ETC-Com genes were suppressed in HHcy conditions and related with Mt complex assembly/activity impairment in human disease and experimental models. We hypothesize that HHcy inhibits Mt complex I gene expression leading to Mt dysfunction.

MP64-16 HYPOFRACTIONATED STEREOTACTIC RADIOTHERAPY INCREASES BOTH TUMOR-INFILTRATING LYMPHOCYTES AND SUPPRESSIVE IMMUNE CELLS IN PROSTATE CANCER

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology II1 Apr 2018MP64-16 HYPOFRACTIONATED STEREOTACTIC RADIOTHERAPY INCREASES BOTH TUMOR-INFILTRATING LYMPHOCYTES AND SUPPRESSIVE IMMUNE CELLS IN PROSTATE CANCER Lin Lin, Nathan Kane, Naoko Kobayashi, Siliva Diaz-Perez, Rajan Kulkarni, Nicholas Nichols, and Robert Reiter Lin LinLin Lin More articles by this author , Nathan KaneNathan Kane More articles by this author , Naoko KobayashiNaoko Kobayashi More articles by this author , Siliva Diaz-PerezSiliva Diaz-Perez More articles by this author , Rajan KulkarniRajan Kulkarni More articles by this author , Nicholas NicholsNicholas Nichols More articles by this author , and Robert ReiterRobert Reiter More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.2061AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Extremely hypofractionated stereotactic body radiotherapy (SBRT) is a technique for delivering radiotherapy (RT) to prostate cancer that is growing in use due to improved convenience as compared to conventional radiotherapy and equivalent efficacy results. It is hypothesized that SBRT may enhance or suppress tumor reactive immunity. The goal of this study is to assess both the anti-tumor and immunosuppressive effects induced by SBRT in prostate cancer patients and experimental models, and ultimately, to test if a combination of SBRT with targeted immune therapy can achieve a superior antitumor effect. METHODS Twelve patients are recruited in our Phase I feasibility trial (NCT02830165) of hypofractionated SBRT followed by radical prostatectomy in high-risk localized prostate cancer. Pre-SBRT biopsies, post-SBRT prostatectomies, and sequential blood samples are collected to assess local radiation-induced tumor immunity and the development of systemic immune response. Syngeneic murine allograft prostate cancer models are used to study the dynamic change of tumor-infiltrating lymphocyte (TIL), T regulatory cell (Treg), myeloid-derived suppressive cell (MDSC), and circulating T memory cell. These immune cell populations from human prostates and experimental models are evaluated using flow cytometry. RESULTS Results from the first patients treated suggest that SBRT increased the density of TILs within the resected prostate tumor two-weeks after treatment, and the majority of the post-SBRT TILs were CD8+ T cells. Co-clinical syngeneic murine prostate cancer models demonstrated a drastic increase in tumor-infiltrating MDSCs immediately after a single RT dose and then a gradual rise of TILs. Besides local effects, irradiation of primary tumors also increased the fractions of MDSCs and Tregs in non-irradiated synchronous tumors, suggesting that hypofractionated SBRT induced a systemic immune response. CONCLUSIONS In prostate cancer patients, hypofractionated SBRT may enhance CD8 T cell infiltration weeks after treatment. In murine prostate cancer models, RT induces an early rise of tumor-infiltrating MDSCs, a gradual increase of TILs, and a systemic immune response in non-irradiated tumors. The expansion of immunosuppressive cell populations induced by SBRT allows us to further explore the effectiveness of SBRT in combination with targeted immune therapies in prostate cancer. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e856 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Lin Lin More articles by this author Nathan Kane More articles by this author Naoko Kobayashi More articles by this author Siliva Diaz-Perez More articles by this author Rajan Kulkarni More articles by this author Nicholas Nichols More articles by this author Robert Reiter More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Three-dimensional experiments and individual based simulations show that cell proliferation drives melanoma nest formation in human skin tissue

BackgroundMelanoma can be diagnosed by identifying nests of cells on the skin surface. Understanding the processes that drive nest formation is important as these processes could be potential targets for new cancer drugs. Cell proliferation and cell migration are two potential mechanisms that could conceivably drive melanoma nest formation. However, it is unclear which one of these two putative mechanisms plays a dominant role in driving nest formation.ResultsWe use a suite of three-dimensional (3D) experiments in human skin tissue and a parallel series of 3D individual-based simulations to explore whether cell migration or cell proliferation plays a dominant role in nest formation. In the experiments we measure nest formation in populations of irradiated (non-proliferative) and non-irradiated (proliferative) melanoma cells, cultured together with primary keratinocyte and fibroblast cells on a 3D experimental human skin model. Results show that nest size depends on initial cell number and is driven primarily by cell proliferation rather than cell migration.ConclusionsNest size depends on cell number, and is driven primarily by cell proliferation rather than cell migration. All experimental results are consistent with simulation data from a 3D individual based model (IBM) of cell migration and cell proliferation.

Population-based mechanistic modeling allows for quantitative predictions of drug responses across cell types

Quantitative mismatches between human physiology and experimental models can be problematic for the development of effective therapeutics. When the effects of drugs on human adult cardiac electrophysiology are of interest, phenotypic differences with animal cells, and more recently stem cell-derived models, can present serious limitations. We addressed this issue through a combination of mechanistic mathematical modeling and statistical analyses. Physiological metrics were simulated in heterogeneous populations of models describing cardiac myocytes from adult ventricles and those derived from induced pluripotent stem cells (iPSC-CMs). These simulated measures were used to construct a cross-cell type regression model that predicts adult myocyte drug responses from iPSC-CM behaviors. We found that (1) quantitatively accurate predictions of responses to selective or non-selective ion channel blocking drugs could be generated based on iPSC-CM responses under multiple experimental conditions; (2) altering extracellular ion concentrations is an effective experimental perturbation for improving the model’s predictive strength; (3) the method can be extended to predict and contrast drug responses in diseased as well as healthy cells, indicating a broader application of the concept. This cross-cell type model can be of great value in drug development, and the approach, which can be applied to other fields, represents an important strategy for overcoming experimental model limitations.

Natural small molecule bigelovin suppresses orthotopic colorectal tumor growth and inhibits colorectal cancer metastasis via IL6/STAT3 pathway

Bigelovin, a sesquiterpene lactone, has been demonstrated to induce apoptosis, inhibit inflammation and angiogenesis in vitro, but its potential anti-metastatic activity remains unclear. In the present study, two colon cancer mouse models, orthotopic tumor allografts and experimental metastatic models were utilized to investigate the progression and metastatic spread of colorectal cancer after bigelovin treatments. Results showed that bigelovin (intravenous injection; 0.3–3 mg/kg) significantly suppressed tumor growth and inhibited liver/lung metastasis with modulation of tumor microenvironment (e.g. increased populations of T lymphocytes and macrophages) in orthotopic colon tumor allograft-bearing mice. Furthermore, the inhibitory activities were also validated in the experimental human colon cancer metastatic mouse model. The underlying mechanisms involved in the anti-metastatic effects of bigelovin were then revealed in murine colon tumor cells colon 26-M01 and human colon cancer cells HCT116. Results showed that bigelovin induced cytotoxicity, inhibition of cell proliferation, motility and migration in both cell lines, which were through interfering IL6/STAT3 and cofilin pathways. Alternations of the key molecules including Rock, FAK, RhoA, Rac1/2/3 and N-cadherin, which were detected in bigelovin-treated cancer cells, were also observed in the tumor allografts of bigelovin-treated mice. These findings strongly indicated that bigelovin has potential to be developed as anti-tumor and anti-metastatic agent for colorectal cancer.

The impact of naloxegol on anal sphincter function - Using a human experimental model of opioid-induced bowel dysfunction

Background and aimsOpioid treatment interferes with anal sphincter function and its regulation during defecation. This may result in straining, incomplete evacuation, and contribute to opioid-induced bowel dysfunction (OIBD). Employing an experimental model of oxycodone-induced OIBD, we hypothesized that co-administration of the peripherally acting μ-opioid antagonist naloxegol would improve anal sphincter function in comparison to placebo. MethodsIn a double-blind randomized crossover trial, 24 healthy males were assigned to a six-day treatment of oral oxycodone 15 mg twice daily in combination with either oral naloxegol 25 mg once daily or placebo. At baseline and at day 6, anal resting pressure and the recto-anal inhibitory reflex (RAIR) were evaluated using manometry and rectal balloon distension. Furthermore, the functional lumen imaging probe was used to measure distensibility of the anal canal. Gastrointestinal symptoms were assessed with the Patient Assessment of Constipation Symptom (PAC-SYM) questionnaire and the Bristol Stool Form Scale. ResultsDuring oxycodone treatment, naloxegol improved RAIR-induced sphincter relaxation by 15% (−45.9 vs −38.8 mm Hg; P < 0.01). No differences in anal resting pressure and anal canal distensibility were found between treatments (all P > 0.5). Naloxegol improved PAC-SYM symptoms (mean score over days; 2.6 vs 4.5, P < 0.001) and improved stool consistency scores (mean score over days; 3.3 vs 2.9, P < 0.01). ConclusionsIn this experimental model of OIBD, naloxegol improved the RAIR and reduced gastrointestinal symptoms. Hence, in contrast to conventional laxatives, naloxegol may regulate opioid-induced anal sphincter dysfunction and facilitate the defecation process.

MUSCLEMOTION: A Versatile Open Software Tool to Quantify Cardiomyocyte and Cardiac Muscle Contraction In Vitro and In Vivo.

There are several methods to measure cardiomyocyte and muscle contraction, but these require customized hardware, expensive apparatus, and advanced informatics or can only be used in single experimental models. Consequently, data and techniques have been difficult to reproduce across models and laboratories, analysis is time consuming, and only specialist researchers can quantify data. Here, we describe and validate an automated, open-source software tool (MUSCLEMOTION) adaptable for use with standard laboratory and clinical imaging equipment that enables quantitative analysis of normal cardiac contraction, disease phenotypes, and pharmacological responses. MUSCLEMOTION allowed rapid and easy measurement of movement from high-speed movies in (1) 1-dimensional in vitro models, such as isolated adult and human pluripotent stem cell-derived cardiomyocytes; (2) 2-dimensional in vitro models, such as beating cardiomyocyte monolayers or small clusters of human pluripotent stem cell-derived cardiomyocytes; (3) 3-dimensional multicellular in vitro or in vivo contractile tissues, such as cardiac "organoids," engineered heart tissues, and zebrafish and human hearts. MUSCLEMOTION was effective under different recording conditions (bright-field microscopy with simultaneous patch-clamp recording, phase contrast microscopy, and traction force microscopy). Outcomes were virtually identical to the current gold standards for contraction measurement, such as optical flow, post deflection, edge-detection systems, or manual analyses. Finally, we used the algorithm to quantify contraction in in vitro and in vivo arrhythmia models and to measure pharmacological responses. Using a single open-source method for processing video recordings, we obtained reliable pharmacological data and measures of cardiac disease phenotype in experimental cell, animal, and human models.

Population-Based Mechanistic Modeling Allows for Quantitative Predictions of Drug Responses across Cell Types

Quantitative mismatches between human physiology and experimental models can be problematic for the development of effective therapeutics. When the effects of drugs on human adult cardiac electrophysiology are of interest, phenotypic differences with animal cells, and more recently stem cell-derived models, can present serious limitations. We addressed this issue through a combination of mechanistic mathematical modeling and statistical analyses. Physiological metrics were simulated in heterogeneous populations of models describing cardiac myocytes from adult ventricles and induced pluripotent stem cells derived cardiac myocytes (iPSC-CMs). These simulated measures were used to construct a cross-cell type regression model that predicts adult myocyte drug responses from iPSC-CM behaviors. We found that: (1) quantitatively accurate predictions of adult myocyte responses to selective or non-selective ion channel blocking drugs could be generated based on iPSC-CM responses under multiple experimental conditions; (2) altering extracellular ion concentrations is an effective experimental perturbation for improving the model's predictive strength; (3) the method can be extended to predict and contrast drug responses in diseased as well as healthy cells, indicating a broader application of the concept. This cross-cell type model can be of great value for drug development, and the approach, which can be applied to other fields, represents an important strategy for overcoming experimental model limitations.

Relationships between vitamin D and paraspinal muscle: human data and experimental rat model analysis

Background ContextVitamin D deficiency (VDD) has been closely linked with skeletal muscle atrophy in many studies, but to date no study has focused on the paraspinal muscle. PurposeTo verify paraspinal muscle changes according to serum vitamin D level. Study DesignA cross-sectional study of patients who visited our hospital and an in vivo animal study. MethodsWe measured serum vitamin D concentration in 91 elderly women and stratified them according to their vitamin D status in three groups, control, vitamin D insufficiency, and VDD, and obtained magnetic resonance imaging data of the lumbar spine and evaluated the quality and quantity of the paraspinal muscles. Additionally, we designed experimental rat models for VDD and VDD replacement. Then, we analyzed the microcomputed tomography data and histologic data of paraspinal muscles, and the histologic data and reverse transcription-quantitative polymerase chain reaction data of intramyonuclear vitamin D receptor (VDR) in paraspinal muscle through comparison with control rats (n=25, each group). This work was supported by a Biomedical Research Institute grant ($40,000), Kyungpook National University Hospital (2014). ResultsIn the human studies, a significant decrease was noted in the overall paraspinal muscularity (p<.05) and increase in fatty infiltration in the VDD group as compared with the other groups (p<.05). In the rat experiment, a decrease was noted in paraspinal muscle fiber size and VDR concentration and VDR gene expression level, and total muscle volume of the VDD rats as compared with the control rats (p<.05). Vitamin D replacement after VDD could partially restore the muscle volume, muscle fiber size, and intramyonuclear VDR concentration levels (p<.05) of the paraspinal muscles. ConclusionsVDD induces paraspinal muscle atrophy and decreases the intramyonuclear VDR concentration and VDR gene expression level in these muscles. Vitamin D replacement contributes to the recovery from atrophy and restoration of intramyonuclear VDR concentration in VDD status.

Human Fetal-Derived Enterospheres Provide Insights on Intestinal Development and a Novel Model to Study Necrotizing Enterocolitis (NEC).

Background & AimsUntreated necrotizing enterocolitis (NEC) can lead to massive inflammation resulting in intestinal necrosis with a high mortality rate in preterm infants. Limited access to human samples and relevant experimental models have hampered progress in NEC pathogenesis. Earlier evidence has suggested that bacterial colonization of an immature and developing intestine can lead to an abnormally high inflammatory response to bacterial bioproducts. The aim of our study was to use human fetal organoids to gain insights into NEC pathogenesis.MethodsRNA sequencing analysis was performed to compare patterns of gene expression in human fetal-derived enterospheres (FEnS) and adult-derived enterospheres (AEnS). Differentially expressed genes were analyzed using computational techniques for dimensional reduction, clustering, and gene set enrichment. Unsupervised cluster analysis, Gene Ontology, and gene pathway analysis were used to predict differences between gene expression of samples. Cell monolayers derived from FEnS and AEnS were evaluated for epithelium function and responsiveness to lipopolysaccharide and commensal bacteria.ResultsBased on gene expression patterns, FEnS clustered according to their developmental age in 2 distinct groups: early and late FEnS, with the latter more closely resembling AEnS. Genes involved in maturation, gut barrier function, and innate immunity were responsible for these differences. FEnS-derived monolayers exposed to either lipopolysaccharide or commensal Escherichia coli showed that late FEnS activated gene expression of key inflammatory cytokines, whereas early FEnS monolayers did not, owing to decreased expression of nuclear factor-κB–associated machinery.ConclusionsOur results provide insights into processes underlying human intestinal development and support the use of FEnS as a relevant human preclinical model for NEC. Accession number of repository for expression data: GSE101531.

A comprehensive molecular and morphological study of the effects of space flight on human capillary endothelial cells: sample quality assessment and preliminary results.

ESA (ESA ILSRA-2009-1026); ASI (contract no. 5681); Regione Toscana (POR FSE 2007-2013-FORTEC); Kayser Italia; Lions Club International, District 108LA, Toscana, Italy.

Multicenter assessment of quantitative sensory testing (QST) for the detection of neuropathic-like pain responses using the topical capsaicin model

ABSTRACT Background The use of quantitative sensory testing (QST) in multicenter studies has been quite limited, due in part to lack of standardized procedures among centers. Aim The aim of this study was to assess the application of the capsaicin pain model as a surrogate experimental human model of neuropathic pain in different centers and verify the variation in reports of QST measures across centers. Methods A multicenter study conducted by the Quebec Pain Research Network in six laboratories allowed the evaluation of nine QST parameters in 60 healthy subjects treated with topical capsaicin to model unilateral pain and allodynia. The same measurements (without capsaicin) were taken in 20 patients with chronic neuropathic pain recruited from an independent pain clinic. Results Results revealed that six parameters detected a significant difference between the capsaicin-treated and the control skin areas: (1) cold detection threshold (CDT) and (2) cold pain threshold (CPT) are lower on the capsaicin-treated side, indicating a decreased in cold sensitivity; (3) heat pain threshold (HPT) was lower on the capsaicin-treated side in healthy subjects, suggesting an increased heat pain sensitivity; (4) dynamic mechanical allodynia (DMA); (5) mechanical pain after two stimulations (MPS2); and (6) mechanical pain summation after ten stimulations (MPS10), are increased on the capsaicin-treated side, suggesting an increased in mechanical pain (P < 0.002). CDT, CPT and HPT showed comparable effects across all six centers, with CPT and HPT demonstrating the best sensitivity. Data from the patients showed significant difference between affected and unaffected body side but only with CDT. Conclusion These results provide further support for the application of QST in multicenter studies examining normal and pathological pain responses.

Chapter 4 - Experimental models of human prion diseases and prion strains

Prion strains occur in natural prion diseases, including prion diseases of humans. Prion strains can correspond with differences in the clinical signs and symptoms of disease and the distribution of prion infectivity in the host and are hypothesized to be encoded by strain-specific differences in the conformation of the disease-specific isoform of the host-encoded prion protein, PrPTSE. Prion strains can differ in biochemical properties of PrPTSE that can include the relative sensitivity to digestion with proteinase K and conformational stability in denaturants. These strain-specific biochemical properties of field isolates are maintained upon transmission to experimental animal models of prion disease. Experimental human models of prion disease include traditional and gene-targeted mice that express endogenous PrPC. Transgenic mice that express different polymorphs of human PrPC or mutations in human PrPC that correspond with familial forms of human prion disease have been generated that can recapitulate the clinical, pathologic, and biochemical features of disease. These models aid in understanding disease pathogenesis, evaluating zoonotic potential of animal prion diseases, and assessing human-to-human transmission of disease. Models of sporadic or familial forms of disease offer an opportunity to define mechanisms of disease, identify key neurodegenerative pathways, and assess therapeutic interventions.

Annurca Apple Nutraceutical Formulation Enhances Keratin Expression in a Human Model of Skin and Promotes Hair Growth and Tropism in a Randomized Clinical Trial.

Several pharmaceutical products have been formulated over the past decades for the treatment of male and female alopecia, and pattern baldness, but relatively few metadata on their efficacy have been published. For these reasons, the pharmaceutical and medical attention has recently focused on the discovery of new and safer remedies. Particularly, great interest has been attracted by oligomeric procyanidin bioactivity, able to promote hair epithelial cell growth as well as to induce the anagen phase. Specifically, the procyanidin B2, a dimeric derivative extracted from apples, has demonstrated to be one of the most effective and safest natural compounds in promoting hair growth, both in vitro and in humans by topical applications. By evaluating the polyphenolic content of different apple varieties, we have recently found in the apple fruits of cv Annurca (AFA), native to Southern Italy, one of the highest contents of oligomeric procyanidins, and, specifically, of procyanidin B2. Thus, in the present work we explored the in vitro bioactivity of AFA polyphenolic extract as a nutraceutical formulation, named AppleMets (AMS), highlighting its effects on the cellular keratin expression in a human experimental model of adult skin. Successively, testing the effects of AMS on hair growth and tropism in healthy subjects, we observed significant results in terms of increased hair growth, density, and keratin content, already after 2 months. This study proves for the first time the impact of apple procyanidin B2 on keratin biosynthesis in vitro, and highlights its effect as a nutraceutical on human hair growth and tropism.