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Phenotypic screens still produce novel drug discoveries, even though the emergence of the finer molecular details of cancer favors target-focused approaches. Recent efforts to improve the design and power of phenotypic screens have addressed the persistent shortcomings of target-focused discovery, such as those related to heterogeneity, resistance, or redundancy. Innovations in culturing patient-derived cancer cells in a tumor-relevant microenvironment have improved the disease relevance of findings. Technological advances for high-throughput screening, ranging from imaging to omics, have increased the information content of readouts and strengthened discovery by phenotype-focused approaches. Before molecular pathways in cancer were known to a depth that could predict targets, drug development relied on phenotypic screening, where the effectiveness of candidate chemicals is judged from functional readouts without considering the mechanisms of action. The unraveling of tumor-specific pathways has brought targets for molecularly driven drug discovery, but precedents in the field have shown that awareness of pathways does not necessarily predict therapeutic efficacy, and many cancers still lack druggable targets. Phenotypic screening therefore retains a niche in drug development where a targeted approach is not informative. We analyze the unique advantages of phenotypic screens, and how technological advances have improved their discovery power. Notable advances include the use of larger biological panels and refined protocols that address the disease-relevance and increase data content with imaging and omic approaches. [ABSTRACT FROM AUTHOR]

Background: To investigate the powerful placebo effects in antidepressant drug trials and their mechanisms, recent pioneering experimental studies showed that expectation manipulation combined with an active placebo attenuated induced sadness. In the present study, we aimed at extending these findings by assessing the psychophysiological response in addition to mere self-report.Methods: One hundred and thirteen healthy female students were randomly assigned to a drug expectation group (active placebo, positive treatment expectation), placebo expectation group (active placebo, no treatment expectation), or a no-treatment group (no placebo, no treatment expectation). After placebo intake, sadness was induced by self-deprecating statements using the Velten method combined with sad music, including a rumination phase. Sadness was measured using the Positive and Negative Affect Schedule Expanded Form (PANAS-X). Heart rate and skin conductance were assessed continuously.Results: After mood induction and after rumination, self-reported sadness was significantly lower, and skin conductance level was significantly higher, in the drug expectation group than in the no-treatment group. The mood induction was further accompanied by a heart rate deceleration within all groups.Limitations: Generalizability is limited by sample selectivity and focusing on sadness as a symptom of depression, exclusively.Conclusion: Expectation-induced placebo effects significantly influenced sadness-correlated changes in autonomic arousal, and not only subjectively reported sadness, indicating that placebo effects in the context of affect are not merely due to subjective response bias. The systematic modification of treatment expectation could be utilized in clinical practice to optimize current therapeutic approaches to improve mood regulation. [ABSTRACT FROM AUTHOR]

Background: Buprenorphine, used for opioid use disorder (OUD) treatment during pregnancy, provides unknown effects on maternal physiological activity. The primary aim of this report is to document acute effects of buprenorphine administration on indicators of maternal autonomic functioning. Effects of maternal buprenorphine dose and other substance exposures on maternal measures were examined, as were neonatal abstinence syndrome (NAS) outcomes.Methods: Forty-nine pregnant, buprenorphine-maintained women yielded maternal physiologic information (heart rate and variability, electrodermal activity, and respiratory rate) at 24, 28, 32 and 36 weeks gestation. Monitoring at trough and peak maternal medication levels was implemented to ascertain acute physiologic effects of buprenorphine administration.Results: Buprenorphine administration accelerated maternal heart rate and reduced variability at two gestational ages (24 and 36 weeks) and suppressed sympathetic (electrodermal) activation at 24, 28 and 32 weeks at times of peak maternal medication levels. Maternal autonomic parameters were unrelated to polysubstance exposure with the exception of cigarette smoking. Heavier smoking dampened maternal heart rate variability across gestation and potentiated reactivity to buprenorphine at 24 and 36 weeks. Heavier smoking was also associated with reduced electrodermal activity at 36 weeks. Buprenorphine dose was unrelated to observed effects. Larger degree of maternal heart rate reactivity to buprenorphine administration was related to more severe NAS expression.Conclusions: These findings detail the maternal autonomic response to buprenorphine administration but also illustrate the significant effect of concurrent cigarette use on maternal autonomic regulation. This suggests the importance of smoking-reduction strategies in the comprehensive, medication-assisted treatment of women with OUD. [ABSTRACT FROM AUTHOR]

Abstract Background Sustained, low- and mid-level (3–6%), radial stretching combined with varying concentrations of sodium pyruvate (NaPy) supplement increase the migration rate during microscale gap closure following an in vitro injury; NaPy is a physiological supplement often used in cell-culture media. Recently we showed that low-level tensile strains accelerate in vitro kinematics during en masse cell migration; topically applied mechanical deformations also accelerate in vivo healing in larger wounds. The constituents and nutrients at injury sites change. Thus, we combine a supplement with stretching conditions to effectively accelerate wound healing. Methods Monolayers of murine fibroblasts (NIH3T3) or myoblasts (C2C12) were cultured in 1 mM NaPy on stretchable, linear-elastic substrates. Monolayers were subjected to 0, 3, or 6% stretching using a custom three-dimensionally printed stretching apparatus, micro-damage was immediately induced, media was replaced with fresh media containing 0, 1, or 5 mM NaPy, and cell migration kinematics during gap-closure were quantitatively evaluated. Findings In myoblasts, the smallest evaluated strain (3%, minimal risk of damage) combined with preinjury (1 mM) and post-injury exogenous NaPy supplements accelerated gap closure in a statistically significant manner; response was NaPy concentration dependent. In both fibroblasts and myoblasts, when cells were pre-exposed to NaPy, yet no supplement was provided post-injury, mid-level stretches (6%) compensated for post-injury deficiency in exogenous NaPy and accelerated gap-closure in a statistically significant manner. Interpretation Small deformations combined with NaPy supplement prior-to and following cell-damage accelerate en masse cell migration and can be applied in wound healing, e.g. to preventatively accelerate closure of microscale gaps. Highlights • Low strain with sodium pyruvate (NaPY) accelerates cell migration in gap closure. • Post-injury NaPy supplement accelerates closure following pre-injury exposure. • Strain of 6% with pre-injury NaPy exposure compensates for lack of post-injury NaPy. • Non-damaging stretch (3 or 6%) accelerates migration of fibroblasts and myoblasts. [ABSTRACT FROM AUTHOR]

Introduction: Antiplatelet medication use continues to rise in an aging population, and these agents can have a deleterious effect for patients with traumatic intracranial hemorrhage (tICH). The purpose of the current investigation is to assess the safety and efficacy of using platelet reactivity testing (PRT) to direct platelet transfusion for tICH patients.Patients and Methods: A Level I trauma center adopted a targeted platelet transfusion guideline using PRT to determine whether platelets were inhibited by an antiplatelet medication (aspirin or P2Y12 inhibitors). Non-inhibited patients were monitored without platelet transfusion, regardless of severity of the head injury. The guideline was analyzed retrospectively to evaluate patient outcomes during the study period (June 2014-December 2016). All patients sustained blunt tICH and received a PRT for known or suspected antiplatelet medication use. Differences were assessed with Kruskal-Wallis and Fisher's Exact tests.Results: 166 patients met study inclusion criteria. PRT results indicated that 48 patients (29%) were not inhibited by an antiplatelet medication, and 92% of those patients (n = 44) were spared platelet transfusion. Seven percent (n = 11) of all patients had a clinically significant progression of the head bleed, but this did not differ by inhibition or transfusion status. Implementation of this guideline reduced platelet transfusions by an estimated 30-50% and associated healthcare costs by 42%.Conclusions: A targeted platelet transfusion guideline using PRT reduced platelet usage for patients with tICH. If appropriately tested, results suggest that not all tICH patients taking or suspected of taking antiplatelet drugs need platelet transfusion. Platelet reactivity testing can significantly reduce healthcare costs and resource usage. [ABSTRACT FROM AUTHOR]

Objectives: To investigate the effect of kallikrein-related peptidase KLK1 on azoospermic mice induced by busulfan and mouse spermatogonial stem cell.Methods: Mice were treated with a single intraperitoneal injection of busulfan, and 4 weeks later, they received a daily intraperitoneal injection of KLK1 at different doses for another 4 weeks. Eight weeks after the busulfan treatment, all mice were sacrificed and their testes were collected for histological evaluation, immunostaining and protein extraction. In vitro, immortalized mouse spermatogonial stem cells, namely C18-4 cells, were treated with KLK1 for proliferation assays.Results: Histological evaluation of testes, epididymis and epididymal fluid showed that KLK1-treated mice had better spermatogenesis than the control group. Immunostaining showed that tissue samples from testes of KLK1-treated mice had more PLZF- and SCP3-positive cells per seminiferous tubule as well as more PNA-positive cells in the seminiferous tubules. Western blots revealed higher expression levels of PCNA in KLK1-treated mice than in control mice. C18-4 cells treated with KLK1 had a higher proliferation rate and higher expression levels of PCNA, Cyclin A and Cyclin E, and the level of phosphorylated ERK2 were increased after KLK1 treatment.Conclusion: Collectively, KLK1 can improve spermatogenesis in azoospermic mice, and KLK1 can promote the proliferation of mouse spermatogonial stem cells via activating ERK1/2 and cell cycle proteins Cyclin A and Cyclin E. This study could offer novel approach and provide new targets for the treatment of azoospermia. [ABSTRACT FROM AUTHOR]

Background: Pain is a common co-morbidity among clinically depressed individuals. We investigated a group of patients who were treated with repetitive transcranial magnetic stimulation (rTMS) for treatment resistant depression (TRD) and who were assessed for severity of both depression and pain at baseline and throughout treatment.Methods: Records of 71 patients treated for TRD with rTMS from 2008 to 2017 were reviewed. Primary outcome measures including depression severity using the Quick Inventory of Depressive Symptomatology (QIDS) and a 0-10 numeric pain rating scale were assessed at baseline and after every 5 sessions throughout the course of 30 treatments.Results: In the total sample, pain improved significantly over the course of treatment. Changes within subjects in QIDS were associated with the changes in pain (p = 0.011). TRD patients with higher pain scores at baseline tended to be older, experienced a longer duration of illness, and showed significant differences in QIDS over treatment time as compared with the low baseline pain group. Patients who had failed a serotonin norepinephrine reuptake inhibitor (SNRI) (venlafaxine or duloxetine) trial in the past had less pain at baseline and showed a group difference in pain scores at all time points, which was significant at treatments 20, 25 and 30, compared to patient groups who had never taken these medications or were currently taking these medications.Limitations: Limitations include the potential impact of the discomfort over the treatment site on the scalp, as it is unclear whether patients' assessment of pain included this side effect, and the lack of a control group due to the naturalistic design of this study.Conclusion: Our data show that pain and depression respond well to rTMS in a TRD population. Pain and depression severity in rTMS patients may be associated over the course of rTMS treatment time-points in individuals with higher levels of baseline pain. [ABSTRACT FROM AUTHOR]

Graphical abstract Abstract The lipid membrane composition defines the physical and pharmacological characteristics of liposomal drugs, and it can be tailored to meet the desired drug delivery needs. The current study is aimed to provide a sharper understanding of the lipid composition effect on doxorubicin (DOX) delivery kinetics, using cholesterol and phosphatidylcholine lipids (PCs) with different acyl chains in liposomal DOX formulations. The PCs were distearoyl (DSPC), dipalmitoyl (DPPC), dimyristoyl (DMPC) and egg-derived PC (EPC), either alone or in combination with cholesterol. Several characteristics were monitored, including DOX loading capacity of liposomes, DOX release in phosphate buffered saline (PBS), PBS/human plasma including buffy coat and human blood, cell uptake, as well as in vivo distribution and therapeutic effects in BALB/c mice bearing C26 colon carcinoma. Addition of cholesterol to liposomal formulation enhanced the particle size stability of the liposomes and the DOX-to-lipid ratio. EPC-liposomes and EPC/Cholesterol-liposomes showed few distinctive features. Overall, cholesterol decreased DOX release from the liposomes, and longer saturated fatty acyl chains in PC decreased DOX release and side-effects and increased the anti-tumor effects of liposomal DOX. [ABSTRACT FROM AUTHOR]

Abstract Background Augmenter of liver regeneration (ALR) protects liver from various injuries, however, the association of ALR with liver fibrosis, particularly its effect on hepatic stellate cells (HSC), remains unclear. In this study, we investigated the impact of ALR on the activation of HSC, a pivotal event in occurrence of liver fibrosis. Methods Liver fibrosis was induced in vivo in mice with heterozygous ALR knockdown (ALR -KD) by administration of CCl 4 or bile duct ligation. The effect of ALR -KD and ALR-overexpression on liver fibrosis was studied in mice and in HSC cells as well. Results Hepatic collagen deposition and expression of α-smooth muscle actin (α-SMA) were significantly increased in the ALR -KD mice compared to wild-type mice. In vitro , ALR -shRNA resulted in the activation of HSC cell line (LX-2). Furthermore, ALR -shRNA promoted LX-2 cell migration, accompanied by increased filamentous actin (F-actin) assembly. The ALR -KD-mediated increase in HSC migration was associated with mitochondrial fusion, resulting in mitochondria elongation and enhancing ATP production. In contrast, ALR transfection (ALR -Tx) decelerated HSC migration and inhibited F-actin assembly, concomitantly enhancing mitochondrial fission and reducing ATP synthesis. Mechanically, stimulation of HSC migration by ALR -shRNA was attributed to the increased mitochondrial Ca2+ influx in HSCs. Treatment of ALR -shRNA-cells with Ruthenium Red (RuR), a specific inhibitor of mitochondrial calcium uniporter (MCU), significantly suppressed mitochondrial Ca2+ influx, HSC migration, mitochondrial fusion and ATP production. ALR -KD-induced HSC migration was verified in vitro in primary mouse HSCs. Conclusion Inhibition of ALR expression aggravates liver fibrosis, probably via promoting HSC migration and mitochondrial fusion. Graphical abstract Unlabelled Image Highlights • Expressions of ALR were evaluated in human fibrotic liver tissue and hepatic tissues in mice with experimental liver fibrosis. • ALR expression was decreased following the activation of HSCs. • Knockdown of ALR deteriorates liver fibrosis and activates HSCs, thereby promoting F-actin assembly and accelerating the migration of HSCs • HSC activation caused by ALR knockdown is linked with elevated mitochondrial calcium and increased ATP synthesis. [ABSTRACT FROM AUTHOR]

Background: There is growing interest in non-invasive brain stimulation techniques as treatments for addictive disorders. While multiple reviews have examined the effects of neuromodulation on craving and consumption, there has been no review of how neuromodulation affects cognitive functioning in addiction. This systematic review examined studies of the cognitive effects of transcranial direct current stimulation (tDCS) and transcranial magnetic stimulation (TMS) in individuals exhibiting addictive behavior.Methods: Articles were identified through searches in PubMed and PsycINFO conducted in October 2017. Eligible studies investigated the effects of tDCS or TMS on cognitive task performance in participants reporting substance use (e.g., alcohol, tobacco, or drugs) or addictive behaviors (e.g., gambling). Tasks were organized into five domains: (1) Inhibitory control, (2) Risk-taking, (3) Impulsive choice (delay discounting), (4) Executive function, and (5) Implicit biases.Results: Twenty-four articles met the inclusion criteria. Fifty-seven percent of studies used tDCS and 43% used TMS, with nearly all studies (96%) targeting the dorsolateral prefrontal cortex. Ten studies reported significant within-subject modulation of cognitive functioning associated with active TMS or tDCS, with the same number reporting no change in cognitive performance. Of four studies that included both an experimental and control participant group, three showed between-group differences in the effects of neuromodulation.Conclusions: While positive effects in several studies suggest that tDCS and TMS improve cognitive functioning in addiction, there is substantial heterogeneity across studies. We discuss person-related and methodological factors that could explain inconsistencies, and propose individualized stimulation protocols may sharpen the cognitive effects of neuromodulation in addiction. [ABSTRACT FROM AUTHOR]

Objective: Naltrexone has been shown to attenuate craving and the subjective effects of methamphetamine. Although naltrexone has modulatory effects on neural activity at dopaminergic synapses, the effect on striatal connectivity is unclear. As methamphetamine use is associated with greater resting-state functional connectivity (RSFC) in the dopaminergic system, we examined whether extended-release naltrexone (XR-NTX) can normalize striatal connectivity and whether changes in RSFC are associated with changes in craving and methamphetamine use.Methods: Thirty-seven participants in or seeking treatment for methamphetamine use disorder took part in this clinical trial at a university-based research clinic between May 2013 and March 2015 (Clinicaltrials.gov NCT01822132). Participants were randomized by a random number generator to a single four-week injection of XR-NTX or placebo. Functional magnetic resonance imaging (fMRI) and self-reported measures of craving and methamphetamine use were conducted before and after double-blinded randomization.Findings: There was a significant reduction in methamphetamine use in the naltrexone group and a significant treatment-by-time interaction on RSFC between the ventral striatum, amygdala, hippocampus, and midbrain. Connectivity was significantly reduced over time in participants randomized to naltrexone but unchanged in those randomized to placebo (p < 0.05, whole-brain corrected). Interactions between treatment and changes in connectivity show that significant reductions in connectivity were associated with reductions in methamphetamine use.Conclusions: Neurobiological deficits associated with methamphetamine use may undermine the efficacy of pharmacotherapies that directly target the dopamine reward system. Naltrexone, via antagonism of indirect mu-opioid effects on dopamine neurons, may attenuate reward system connectivity and aid in methamphetamine use treatment. [ABSTRACT FROM AUTHOR]

Highlights • The models of subchronic acrylamide (ACR) poisoning and calpeptin (CP) intervention have been set up. • CP has an intervening effect on ACR-induced MT injury in sciatic nerve. • The mechanism of ACR-induced MT injury is associated with the channel of calpain- PKC/CDK5-MAP2/Tau-MTs. Abstract The present study aimed to investigate the protective effect and mechanism of calpeptin (CP) on acrylamide (ACR)-induced microtubule (MT) injury in the sciatic nerve of rats. All rats were divided into four groups (control, CP, ACR, and ACR + CP):1 ml/kg saline, 200 μg/kg CP, 30 mg/kg ACR, and 30 mg/kg ACR plus 200 μg/kg CP were administered to the corresponding rats for 4 weeks through intraperitoneal injection. Body weight and neurobehavioral indicators were measured weekly and α-tubulin, β-tubulin, and other concerned proteins were estimated by western blotting and immunohistochemistry. At 4 weeks, decreased body weight, increased gait scores, increased hindlimb splay, and decreased time of fall of ACR rats were observed compared with those of control rats. All these mentioned changes were restored in the ACR + CP group compared with the ACR group. After 4 weeks of administration, western blotting and immunohistochemistry revealed significant increase in the protein levels of β-tubulin, calpain I, calpain II, Tau, microtubule-associated protein 2 (MAP2), protein kinase C, and cyclin-dependent kinase 5 in the ACR group compared with the control group; these increases were significantly lower in the ACR + CP group than in the ACR group. Furthermore, histopathological examination revealed loose arrangement, disorganised structure, uneven density, and exfoliated perineurium in the ACR group, and CP administration improved these changes significantly. The present results suggest that CP has an intervening effect on ACR-induced MT injury. A possible mechanism is that calpain maintains the stability of MTs by regulating the metabolism of Tau and MAP2. [ABSTRACT FROM AUTHOR]

Abstract MicroRNA-146a is a well-studied microRNA participating in immune and inflammatory diseases, but its role in sepsis has not been investigated. Here in our study, we found increased level of microRNA-146a in macrophage stimulated by lipopolysaccharide. In addition, the mRNA level of Notch1 was also increased. Up-regulation of microRNA-146a by miR-146a-mimic alleviated inflammatory responses of macrophage, for the levels of IL-1β, IL-6 and CCL-2 were decreased, and the activation of NF-κB signaling was inhibited. The histological examination showed that microRNA-146a protected against organ damage in mice with lipopolysaccharide injection, and the level of inflammatory factors, Cr, BUN, AST and ALT in serum were all decreased, reflecting the alleviated inflammation and recovered organ function. Then predicting databases were used and Notch1 was predicted as one of the potential targets of microRNA-146a. Knockout of Notch1 in macrophage showed reduced secretion of inflammatory factors and attenuated activation of NF-κB signaling in response to lipopolysaccharide. Specifically knockout of Notch1 in macrophage protected mice from LPS induced organ damage and dysfunction. Therefore, we prove that miR-146a acts as an inhibitor of inflammation by targeting Notch1 in macrophage, therefore protects mice from organ damage in sepsis. Highlights • LPS increased miR-146a in macrophage. • miR-146a decreased inflammatory response of macrophage and protected against organ damage induced by LPS. • miR-146a directly targeted Notch1 to anti-inflammation. [ABSTRACT FROM AUTHOR]

Abstract We herein report a unique case of type B2 thymoma-associated myasthenia gravis which was ameliorated by immunosuppressive therapy in combination with chemotherapy. However, the patient subsequently developed pure red cell aplasia and marked lymphocytosis after additional chemotherapy aimed at improvement of thymoma. While a separate immunosuppressive regimen was effective for anemia, lymphocytosis was exacerbated. The biopsied thymoma specimen contained CD4+, CD8+, and CD4+/CD8+ T cells, some of which were CD3−, suggesting immature thymocytes. In contrast, majority of the peripheral lymphocytes were polyclonal CD3+/CD8+/T cell receptor (TCR)αβ+ T cells. The CD4/CD8 ratio in the present patient might be affected by immunosuppressive agents, resulting in CD8+ T cell expansion associated with pure red cell aplasia. Although several cases of thymoma accompanied by peripheral T cell lymphocytosis were reported, marked CD8+ T cell proliferation is extremely rare. Highlights • We reported a thymoma with MG, later developed PRCA and CD8+ T cell expansion. • They were polyclonal mature CD8+ T cells, not neoplastic nor thymoma-related T cells. • T cells were expanded after immunosuppressive therapy with steroids and tacrolimus. • Lymphocytosis was worsened after chemotherapy that could reduce thymoma volume. • Thymoma with MG accompanied by marked CD8+ T cell proliferation is extremely rare. [ABSTRACT FROM AUTHOR]

Abstract Dendritic cells (DCs) are antigen-presenting cells with the ability to activate naïve T cells and direct the adaptive cellular immune response toward a specific profile. This is important, as different pathogens demand specific “profiles” of immune responses for their elimination. Such a goal is achieved depending on the maturation/activation status of DCs by the time of antigen presentation to T cells. Notwithstanding this, recent studies have shown that DCs alter their metabolic program to accommodate the functional changes in gene expression and protein synthesis that follow antigen recognition. In this review, we aim to summarize the data in the literature regarding the metabolic pathways involved with DC phenotypes and their functions. Highlights • Metabolism directs the activity of DCs into immunogenic or tolerogenic profile. • Glycolysis is increased in DCs during TLR activation, whereas OXPHOS is decreased. • Rapamycin, AMPK agonists and Rosiglitazone may direct DCs to a tolerogenic phenotype. • PI3K/AKT signaling pathway prevents excessive inflammation. • The modulation of DC metabolism may be used for the development of new drugs. [ABSTRACT FROM AUTHOR]

Abstract Necrotizing enterocolitis (NEC) is a life-threatening condition that can occur in about 7% of pre-term infants, and approximately 20% to 30% of the cases will end in death. An overactive immune response is thought to be a primary instigator of many symptoms during NEC. Hence, we hypothesized that NEC patients might present impairment in regulatory T (Treg) cells that limited their capacity to contain the excessive inflammation-induced damage. To investigate this, peripheral blood mononuclear cells were collected from NEC and non-NEC infants with matching age and weight. Treg cells, identified as CD3+CD4+CD25+/hiFoxp3+ T cells, were present at significantly lower frequency in NEC infants than in non-NEC infants. We also observed that the frequency of IL‑17+ CD4+ T cells was significantly higher in NEC infants, while the frequencies of IL‑10+ and TGF‑β+ CD4+ T cells were significantly lower in NEC infants. The CD4+CD25+/hi Treg cells from NEC infants were capable of suppressing CD4+CD25− T conventional cell proliferation, but with significantly reduced potency than the CD4+CD25+/hi Treg cells from non-NEC infants. In addition, the CD4+CD25+/hi Treg cells from non-NEC infants, but not those from NEC infants, were capable of suppressing IL‑17 expression. Furthermore, the CD4+CD25+/hi Treg cells from NEC infants displayed reduced expression of CTLA‑4, LAG‑3, and Helios, compared to those from non-NEC infants. Overall, these results demonstrated that Treg cells from NEC infants displayed a multitude of functional impairments, and suggested that Treg cells might serve as a treatment target in NEC. Highlights • Frequency of Tregs was significantly lower in NEC infants than in non-NEC infants. • IL‑17+ CD4+ T cells were higher while TGF-β+ CD4+ T cells were lower in NEC infants. • Tregs from NEC were less capable to suppressing CD4+CD25− Tconv proliferation. • Tregs from NEC infants were incapable of suppressing IL‑17 expression. • Tregs from NEC infants displayed reduced expression of CTLA‑4, LAG‑3, and Helios. [ABSTRACT FROM AUTHOR]

Abstract Natural killer (NK) cells eliminate cancer cells in a contact-dependent manner. However, how NK cells find cancer cells remain unclear. Here, using time-lapse imaging, we investigated how individual NK cells migrate toward cancer cells. Although naïve B16F10 cancer cells produce low levels of chemokines, IFN-γ-treated B16F10 cells secreted high levels of CXCL10, low levels of CCL5, but did not secrete CCL2, CCL7, or CXCL12. Wild-type NK cells migrated well toward cancer cells and killed them, whereas NK cells deficient in CXCR3 did not. CXCR3-deficient NK cells also showed slower migration speed than did wild-type NK cells. Taken together, our data show that NK cells find cancer cells, at least in part, by sensing CXCL10 produced by cancer cells and suggest that a strategy to increase CXCL10 secretion by cancer cells may improve the efficacy of NK cell–based immunotherapy. Highlights • Time-lapse imaging reveals how NK cells find cancer cells. • IFN-γ activates cancer cells to produce high levels of CXCL10. • CXCL10 increases migration directionality and speed of wild-type NK cells, but not CXCR3−/− NK cells. [ABSTRACT FROM AUTHOR]

Oxidative stress plays a major role in the development and progression of diabetic nephropathy (DN). In this study, the potential protective effect of ginger ( Zingiber officinale ) rhizome extract on hyperglycemia-induced oxidative stress, inflammation and apoptosis was investigated. An experimental diabetic rat model was induced by intraperitoneal injection of streptozotocin. Diabetic rats were treated orally with 400 or 800 mg/kg/day Z. officinale extract for six weeks. Diabetic animals exhibited elevated blood glucose levels and glycated hemoglobin (HbA1c) with altered lipid profile. Blood urea nitrogen, serum creatinine and urea, and urine albumin levels were significantly increased in diabetic rats. Treatment with Z. officinale ameliorated hyperglycemia, hyperlipidemia and kidney function. In addition, Z. officinale minimized the histological alterations in the kidney of diabetic rats. Chronic hyperglycemia resulted in a significant increase in malondialdehyde, protein carbonyl, pro-inflammatory cytokines, cytochrome c and caspase-3 in the kidney of rats. Z. officinale extract significantly attenuated oxidative stress, inflammation and apoptosis, and enhanced antioxidant defenses in the diabetic kidney. In conclusion, this study strongly suggests that Z. officinale rhizome extract exerts a protective role against diabetes-induced renal injury by ameliorating oxidative stress, inflammation and apoptosis. [ABSTRACT FROM AUTHOR]

Abstract Introduction Obstructive sleep apnea (OSA) is now a more common indication for tonsillectomy than recurrent tonsillitis (RT) [1,2]. Few studies have addressed possible differences in pathogenesis between these two conditions. Children with RT and OSA are often being treated in the community with multiple courses of antibiotics before surgery. Current understanding of the role of bacteria in disorders of the tonsils is mainly based on the culture of tonsil swabs. Swab cultures reflect only a very small fraction of the bacteria present on the mucosal surface and may not represent the bacterial communities within the tonsil crypts [3,4]. This study aimed to evaluate the local lymphocyte response and associations with bacterial community composition using molecular techniques of the tonsils removed from children for RT or OSA. Method The palatine tonsils were removed by extracapsular dissection from 24 patients with age range one to ten years, 14 of whom had RT and 10 had OSA. The fixed tonsil tissues were evaluated for bacteria by Gram-staining and presence of connective tissue by safranin staining. B lymphocytes and T lymphocytes were also measured immunohistochemically. Finally, previously published bacterial community data for this cohort were reassessed for associations with RT and OSA, and with the observed lymphocyte patterns. Results In tonsils from patients with RT, large micro-colonies of bacteria were observed in the tonsil crypts, and a large number of B and T lymphocytes were noted immediately adjacent to the tonsil crypt itself. In marked contrast, the tonsils from patients with OSA had no bacteria identified, and no significant skewing of lymphocytes based on site (such as follicles or crypts). We observed that the majority of lymphocytes surrounding the bacterial micro-colonies were B lymphocytes with a mean ratio of 109:55 (B lymphocytes: T lymphocytes). Bacterial community diversity was not different between the two cohorts; however, there were significant differences in bacterial community composition. Children with RT had a higher relative abundance of members from the genera Parvimonas , Prevotella, and Treponema. While children with OSA had a higher relative abundance of Haemophilus , and Capnocytophaga. Conclusion These results demonstrate significant differences in the local lymphocyte response and bacterial community composition in tonsil tissue between RT and OSA patients. It suggests that the response to antibiotics used in the treatment of these two conditions may be different. Furthermore, the presence of lymphocytes in RT within the tonsil crypt outside the tonsil epithelium is a unique observation of the location of these cells. [ABSTRACT FROM AUTHOR]

Highlights • Peroxidation, increased ROS and oxidative stress can lead to premature aging. • Estrogens and genistein have cytoprotective effects in skin by modulating peroxidation. • Protective effects on skin involve nitric oxide release and mitochondria function. Abstract Background Estrogens and phytoestrogens can hinder the aging process through mechanisms related to estrogen receptors (ERs), guanine nucleotide-binding protein-coupled receptor (GPER30), mitochondria function and nitric oxide (NO) release. Up to date, however, the above issues are a matter of debate. Objective To examine the effects elicited by 17 β-estradiol and genistein against peroxidation in human keratinocytes/fibroblasts and evaluate the role played by ERs, GPER30, mitochondria and NO. Methods Human fibroblasts/keratinocytes, either subjected to peroxidation or not, were exposed to 17 β-estradiol/genistein in the absence or presence of the NO synthase (NOS) inhibitor, the ERs and GPER30 blockers, fulvestrant and G15, the phosphatidyl-inositol-3-kinase (PI3K-Akt), the p38 mitogen-activated protein (MAP) kinase and the extracellular signal–regulated kinases (ERK) 1/2 inhibitors. Specific kits were used for cell viability, NO, ROS and glutathione (GSH) detection and mitochondrial membrane potential measurement. Western Blot analysis was performed for kinases expression/activation detection. Results In physiological and peroxidative conditions, 17 β-estradiol/genistein respectively increased and reduced NO release by fibroblasts/keratinocytes. Moreover, both agents prevented the ROS release and the fall of cell viability and mitochondrial membrane potential, while increasing GSH levels and the proliferation rate. Fulvestrant and G15 counteracted all above responses. Also, the NOS, and the kinases blockers reduced the protection exerted by 17 β-estradiol/genistein on cell viability/mitochondria function. The involvement of PI3K-Akt and p38-MAPK was confirmed by Western blot. Conclusion 17 β-estradiol/genistein protected fibroblasts/keratinocytes against peroxidation by modulating oxidant/antioxidant system and mitochondria membrane potential, through mechanisms related to ERs and GPER30 and kinases activation. [ABSTRACT FROM AUTHOR]

Plasmodium vivax is the most widely spread species causing human malaria. The control of malaria caused by P. vivax has been largely hampered by its ability to develop a dormant liver stage that can generate a new blood infection at different periods of time. Unfortunately, the mechanisms of dormancy in P. vivax have not been thoroughly elucidated to date. In this study, the putative dormancy genes were analyzed to select genes with less genetic variability to maintain the function of relapsing. Expression data concerning these genes were searched to support the selection. Protein interactions among selected gene products were identified based on known and predicted protein-protein interaction using String database. Potentially interacting proteins (n = 15) were used to propose a mechanism involved in dormancy based on the differential vesicular transport due to the iron available in the hepatocyte. [ABSTRACT FROM AUTHOR]

Zinc (Zn) is a fundamental trace element for cell viability and physiology, and concentrations above the physiological level may lead to cell damage. In order to explore the effect of zinc chloride (ZnCl 2 ) on murine photoreceptor cells, in this study, we investigated the alterations of murine photoreceptor cell proliferation and cell morphology after exposure to various concentrations of ZnCl 2 , determined the levels of hydrogen peroxide, hydroxyl radicals, cytochrome c , and ATP before and after cells exposure to different concentrations of ZnCl 2 , monitored the changes of mitochondrial membrane potential, and further explored the expressions of BCL2-associated X (Bax) and B cell CLL/lymphoma (Bcl)-2 at gene and protein levels. The results indicated that appropriate ZnCl 2 levels can enhance the cell proliferation, whereas high levels of ZnCl 2 could apparently inhibit cell growth, exaggerate the generation of both hydrogen peroxide and hydroxyl radicals, collapse the mitochondrial membrane potential, and accelerate cytochrome c release into cytosol, decrease the ATP production, elevate the Bax production, and reduce the Bcl-2 expression, thereby disrupting the mitochondrial homeostasis. Consequently, the disrupted mitochondrial homeostasis initiates the mitochondria-mediated apoptotic signaling pathway, leading to cell death. Taken together, the results suggest that the over generation of reactive oxygen species and the activated mitochondrial signaling pathway play an important role in ZnCl 2 -induced murine photoreceptor cell death. [ABSTRACT FROM AUTHOR]

Highlights • Castration of bull calves increased pain indicator substance P. • Castration increased inflammation indicator haptoglobin. • A lidocaine-plus-flunixin (LF) treatment tended to reduce inflammation indicator. • LF treatment did not reduce elevated cortisol and substance P in castrated calves. • Additional treatment would be beneficial to pain relief in castrated calves. Abstract The aim of this study was to determine the effects of castration and a lidocaine-plus-flunixin (LF) treatment on growth and indicators of pain, inflammation, and liver function in Korean cattle bull calves. Forty Korean cattle bull calves (body weight 197.0 ± 2.94 kg and age 6.3 ± 0.09 months) were each assigned to one of four treatments (n = 10 heads/group): no castration with no LF injection (NC-NLF); no castration with LF injection (NC-LF); castration with no LF injection (C-NLF); and castration with LF injection (C-LF). LF treatment included a local anesthetic lidocaine hydrochloride injection (12 mL of 2% in the scrotum) and intravenous injection of a non-steroidal anti-inflammatory drug, flunixin meglumine (2 mg/kg body weight of 50 mg/mL solution), immediately prior to castration. For the NLF groups, a 0.9% NaCl placebo solution was used. Castration was performed surgically using a Newberry knife and a Henderson castrating tool. Blood was collected immediately before castration and at h 0.5, h 6, d 1, d 3, d 7, and d 14 after castration. Feed intake was recorded daily, and body weight was measured on the day prior to the experiment and at d 14 after castration. Castration tended (P = 0.07) to decrease average daily weight gain, but LF treatment did not affect weight gain. Castration increased both circulating cortisol concentrations (P < 0.001) at h 0.5 after castration and substance P (SP) concentrations (P = 0.001) at h 6 after castration. However, the LF treatment did not significantly reduce cortisol and SP concentrations in castrated animals. Castration increased (P < 0.001) circulating haptoglobin (Hp) concentrations on d 1 and d 3 after castration, and LF treatment tended (P = 0.09) to decrease Hp concentrations on d 1 and decreased (P = 0.02) Hp concentrations on d 3. Castration did not affect glutamic oxaloacetic transaminase (GOT) and glutamic pyruvate transaminase (GPT) concentrations. LF treatment increased GOT concentrations at h 6 (P < 0.001), d 1 (P < 0.001), and d 3 (P = 0.003). LF treatment also increased GPT concentrations on d 1 (P = 0.006) and d 3 (P = 0.003). In conclusion, castration of bull calves resulted in time-sequential increases in circulating concentrations of cortisol at h 0.5, SP at h 6, and Hp on d 1. LF treatment did not significantly reduce elevated cortisol and SP, but tended to decrease elevated Hp concentrations in castrated animals. Our study demonstrates that LF treatment is not sufficient for the reduction of the indicators of pain and inflammation in castrated calves, suggesting that additional alleviation methods are required. [ABSTRACT FROM AUTHOR]

Background Given the changes in the patterns of muscular activation and scapular movement in individuals with subacromial pain syndrome, the use of neuromuscular training has been considered in rehabilitation protocols. There is currently no evidence of the effects of the use of three-dimensional (3D) kinematic biofeedback on individuals with subacromial pain syndrome. This study aimed to determine the immediate effect of scapular motor control exercises using 3D kinematic biofeedback on the scapular kinematics, inter-segment coordination and pain of individuals with subacromial pain syndrome. Method The kinematics of the scapulothoracic joint of 26 subjects with subacromial pain syndrome were assessed in the movement arm elevation and lowering in the sagittal plane before and after performance of three scapula-focused exercises using kinematic biofeedback. The individuals were familiarized with the selected exercises to acquire a greater scapular posterior tilt, while kinematic biofeedback, with visual and auditory stimuli, was used in real time. Scapular kinematics, pain, and subjective perception of exertion were the pre- and post-test measures. Findings In the movement of arm elevation and lowering, no differences were found in scapular tilt and on coordination between the segments pre- and post-test and the effect size was considered small. Interpretation Our results demonstrate that the performance of scapula-focused exercises using kinematic biofeedback does not cause immediate changes in the magnitude of scapular movement. However, inter-segmental coordination showed evidence of changes for scapular tilt in the lowering of the arm and internal rotation in the elevation and the lowering of the arm in individuals with subacromial pain syndrome. [ABSTRACT FROM AUTHOR]

Dasatinib, a new tyrosine kinase inhibitor, is used clinically to kill chronic myelogenous leukemia and acute lymphoblastic leukemia through apoptosis. Obviously, anemia is developed in many patients receiving dasatinib for treatment. Until now, the mechanism for the cytotoxic effects of dasatinib in human erythrocytes is not fully understood. As many tyrosine kinases are found in human erythrocytes, it is therefore logical to hypothesize that dasatinib is able to induce apoptosis (or eryptosis) in human erythrocytes. True to our expectation, dasatinib inhibited tyrosine kinase and induced eryptosis in human erythrocytes with early denature of esterase, cell shrinkage, loss of membrane integrity with inside-out phosphatidylserine, increase in the cytosolic Ca 2+ ion concentration ([Ca 2+ ]i), caspase-3 activation and change in cellular redox state. Mechanistically, the rise of [Ca 2+ ]i seems to be a key mediator in the dasatinib-mediated eryptosis because depletion of external Ca 2+ could suppress the eryptotic effects. Also, dasatinib was able to reduce membrane fluidity in human RBCs. For the direct action on membrane, dasatinib permeabilized RBC ghosts in a way similar to digitonin. Taken together, we report here for the first time that dasatinib inhibited tyrosine kinase and induced eryptosis in human erythrocytes through Ca 2+ loading and membrane permeabilization. [ABSTRACT FROM AUTHOR]

Arsenic, an established human carcinogen, causes genetic toxicity. However, the molecular mechanisms involved remain unknown. MicroRNAs (miRNAs) are regulators that participate in fundamental cellular processes. In the present investigation, we selected, as research subjects, patients with arsenic poisoning caused by burning of coal in Guizhou Province, China. For these patients, the plasma levels of miR-145 were up-regulated. In L-02 cells, arsenite, an active form of arsenic, induced up-regulation of miR-145 and down-regulation of ERCC1 and ERCC2, and caused DNA damage. For L-02 cells, transfection with an miR-145 inhibitor prevented arsenite-induced DNA damage and decreased ERCC2 levels. Luciferase reporter assays showed that miR-145 regulated ERCC2 expression by targeting the 3′-UTR of ERCC2, but not that for ERCC1. The present results demonstrate that arsenite induces the over-expression of miR-145 and inhibits DNA repair via targeting ERCC2, thus promoting DNA damage. The information provides a new mechanism for arsenic-induced liver injury. [ABSTRACT FROM AUTHOR]

Highlights • Resveratrol prevents deficits in spatial reference memory following ethanol administration. • Resveratrol reverses EtOH-induced TLR2 activation in microglia. • Resveratrol mitigated activation of the TLR2/MyD88/NF-κB pathway. Abstract Excessive ethanol (EtOH) intake affects cognitive function and leads to permanent learning and memory deficits. EtOH-induced neuroinflammation plays an important role in EtOH neurotoxicity. Studies have shown that EtOH activates microglia and induces an inflammatory response. Resveratrol (Rsv) is a natural polyphenol found in a wide variety of plants and fruits, and produces the neuroprotective and anti-inflammatory effects in the central nervous system. However, effect of Rsv on EtOH-induced neuroinflammation is still unknown. We investigated the anti-inflammatory effect of Rsv in the context of EtOH-induced neurotoxicity and the molecular mechanisms potentially involved in the effect. The results showed that treatment of rats with Rsv prevented the deficits of spatial reference memory induced by EtOH and mitigated EtOH-induced neuroinflammation by inhibiting microglial activation and decreasing the levels of inflammatory cytokines including interleukin-1β, interleukin-6 and tumor necrosis factor α. The further studies indicated that Rsv reduced TLR2 expression in vivo and in vitro, and downregulated expression of myeloid differentiation primary response 88 (MyD88) and phosphorylation of nuclear factor κB (NF-κB). These data demonstrate that Rsv alleviates the ethanol-induced neuroinflammation via inhibition of TLR2-MyD88-NF-κB signal pathway. [ABSTRACT FROM AUTHOR]

Highlights • Desynchronizing electrical stimulation may be used to treat epilepsy. • Biphasic bilateral asynchronous stimulation has the greatest anticonvulsant effect. • Morphology, spatial and timing parameters display additive effects. Abstract A promising alternative for the treatment of refractory epilepsy is electrical stimulation (ES) of the central nervous system. Based on the premise that epilepsy is a result of neural hypersynchronization, we have previously demonstrated that a novel non-standard form of electrical stimulation with randomized inter-pulse intervals, termed non-periodic stimulation (NPS), applied to the amygdala is robustly anticonvulsant. This investigation also suggested that NPS attains its therapeutic effect by desynchronization of epileptiform activity. Here, we further explored the desynchronization hypothesis by testing how the efficacy of NPS in the suppression of convulsive behaviors depends on morphological, spatial, and temporal parameters of stimulus. For this purpose, we varied the number of pulse phases (monopolar versus bipolar square pulses), side of stimulation (right versus left), number of application hemispheres (unilateral versus bilateral), and interhemispheric temporal synchronicity (synchronous versus asynchronous), while measuring the impact on the anticonvulsant action of NPS. Wistar rats received a controlled infusion of the convulsant agent pentylenetetrazole (PTZ, 10 mg/min), together with one of six variations of NPS applied to the amygdala. A stimulated PTZ-free group of animals was also performed as a positive control. Latency to convulsive behavior was used to measure seizure threshold. Animals receiving NPS displayed significant higher threshold for forelimb clonus and generalized tonic-clonic seizures for all patterns. Thresholds seemed to increase gradually from mono to biphasic, unilateral to bilateral, and synchronous to asynchronous stimuli. Thus, combined biphasic, bilateral, and asynchronous stimulation resulted in the greatest seizure threshold. PTZ free animals did not develop any observable convulsive behavior or other uncommon motor activity. These results confirm that NPS has anticonvulsant properties and that biphasic, bilateral, and asynchronous stimulation enhances its efficacy. The fact that lack of synchronism between stimuli of each hemisphere maximizes NPS anticonvulsant power is evidence to desynchronization as tool for suppression of seizures. [ABSTRACT FROM AUTHOR]

Abstract The impact of vanadium (V) and magnesium (Mg) as sodium metavanadate (SMV, 0.125 mg V/ml) and magnesium sulfate (MS, 0.06 mg Mg/ml) on lipid peroxidation (LPO) and selected elements in the rat erythrocytes (RBCs) was investigated. Relationships between some indices determined in RBC were also studied. SMV alone (Group II) elevated the malondialdehyde level (MDA RBC) (by 95% and 60%), compared with the control (Group I) and MS-supplemented rats (Group III), respectively, reduced the concentration of Cu RBC (by 23.5%), in comparison with Group I, but did not change the levels of Na RBC , K RBC , and Ca RBC , whereas MS alone (Group III) only reduced the Cu RBC concentration (by 22%), compared with Group I. The SMV + MS combination (Group IV) reduced and elevated the Cu RBC (by 24%) and Ca RBC (by 111%) concentrations, respectively, in comparison with Groups I and III, and these changes were induced by the V-Mg antagonistic and synergistic interaction, respectively. The combined SMV + MS effect also enhanced the MDA RBC level, compared with Groups I (by 79%) and III (by 47%) and slightly limited its concentration, compared with Group II, which, in turn, resulted from the distinct trend toward the V-Mg antagonistic interaction. We can conclude that V (as SMV) is able to stimulate LPO in rat RBCs and that V-Mg interactive effects are involved in changes in Cu RBC , Ca RBC , and MDA RBC. Further studies are needed to elucidate the exact mechanisms of the V-Mg antagonistic/synergistic interactions and to provide insight into the biochemical mechanisms of changes in rats suffering from anemia [1], characterized by a disrupted antioxidant barrier in RBCs [2] and an intensified free radical process in these cells. Graphical abstract This report provides new information on the effect of vanadate and magnesium and their interactions on some oxidative stress markers and level of selected elements in rat RBCs. Image 1 Highlights • Lipid peroxidation and some elements were investigated in the rat erythrocytes during V and/or Mg administration. • Mg alone did not alter the MDA RBC level but significantly decrease the Cu RBC concentration. • SMV alone markedly increased MDA RBC and decreased the Cu RBC level. • The V-Mg interactive effects were involved in changes in the Cu RBC , Ca RBC , and MDA RBC levels. • The V-Mg antagonistic and synergistic interactions were revealed. [ABSTRACT FROM AUTHOR]

Bone scaffolds should exhibit antimicrobial activity against bacteria, which is one of the problems in the treatment and regeneration of bone tissue in orthopedics and dentistry. Chlorhexidine (CHX) is very common in practical medicine as an antibacterial agent. We have developed a composite biomaterial based on hydroxyapatite (HA), sodium alginate (Alg) and CHX, which can be used as carrier system for local drug delivery, in particular for dental application. The suitability of HA/Alg/CHX composite to act as slow release drug delivery systems was evaluated in phosphate-buffered saline (PBS) using a high-performance liquid chromatography. The dependence of drug release from the content of Alg and the method of the material preparation (drying at 37 °C, lyophilization at −55 °C and annealing at 1100 °C, in form ceramic scaffolds) was investigated. The presence of Alg in the composite increased the volume of adsorbed and released CHX by 2 times, and the release time from 24 to 72 h. The highest concentration of drug in the PBS after 72 h (0.04 mg/l) and the maximum rate of its release (0.83 μg/h) was in the case of lyophilized and lowest (0.015 mg/l and 0.2 μg/h, respectively) in the case of dried samples at 37 °C. [ABSTRACT FROM AUTHOR]

Single instillation (SI) reduces recurrence of non-muscle-invasive bladder cancer by chemoresecting floating tumor cells and residual tumor lesions (RTLs) after transurethral resection of the bladder, but with limited efficacy. Current studies improved this by prolonging retention time and increasing penetration to bladder wall, ignoring that the two separate factors should be treated in different ways. Here, we introduced a smart gel system-based SI to prevent re-implantation of tumor cells (RTCs) and ablate RTLs in a “divide-and-rule” approach. The gel system was synthesized by PEG-PAMAM and dextran aldehyde and composed of gold nanorods and gemcitabine for photothermal therapy and chemotherapy, respectively. It was developed to provide dextran aldehyde-selective adhesion with tissue amines. Since tumor surface expressed high levels of collagen, the exposed amines could act as adhesion points for the gel system. Thus, the gel presented more affinity to tumor tissues. When being instilled, it could form a protective layer on the inner face of the entire bladder wall immediately, preventing RTCs in early time. And it persisted long at the tumor site, ablating RTLs. Our data proved the gel system improved intravesical treatment efficacy in a “divide-and-rule” approach and might be a promising treatment strategy for SI. [ABSTRACT FROM AUTHOR]

Oxidative stress contributes to the initiation and progression of liver damage. SIRT3 is a member of nicotinamide adenine dinucleotide-dependent deacetylases that plays a key role in anti-oxidative defense and mitochondrial function in the liver. Honokiol is a natural lignan from the plants of Magnolia genus that exhibits potent anti-oxidative property. This study aims to evaluate the hepatoprotective potential of honokiol against oxidative injury in tert -butyl hydroperoxide ( t -BHP)-injured AML12 hepatocytes in vitro and carbon tetrachloride (CCl 4 )-stimulated liver damaged mice in vivo and to determine whether or not this effect occurs by activating SIRT3. The results showed honokiol protects t -BHP-injured AML12 hepatocytes and CCl 4 -stimulated liver damage in mice by activating SIRT3. Honokiol reduces the acetylation level of superoxide dismutase 2 to enhance its anti-oxidative capacity, which decreases reactive oxygen species accumulation in AML12 cells. Honokiol increases the deacetylated peroxisome proliferator-activated receptor γ coactivator 1-α level to promote mitochondrial biogenesis. Moreover, honokiol attenuates t -BHP induced mitochondrial fragmentation through Ku70-dynamin-related protein 1 axis. These results suggest that honokiol can ameliorate oxidative damage in hepatocytes by activating SIRT3, which might be a potential therapeutic agent for liver oxidative injury. [ABSTRACT FROM AUTHOR]

It seems that histamine release in the site of neuronal injury could contribute to the neuropathic pain mechanism. In the present study, we investigated the anti-allodynic effects of chronic administration of different classes of histamine H 1 and H 2 receptor antagonists on neuropathic nociceptive behavior following tibial nerve transection (TNT) in rats. Peripheral neuropathy was induced by TNT surgery. We performed acetone tests (AT) to record cold allodynia, Von Frey tests (VFT) to measure mechanical allodynia, double plate test (DPT) to evaluate thermal place preference/avoidance and open field test (OFT) for evaluation of animal activity. TNT rats showed a significant mechanical and cold allodynia compared to the sham group. Chlorpheniramine (5 and 15 mg/kg, i.p) significantly attenuated cold allodynia and prevented cold plate avoidance behavior and at the dose of 15 mg/kg remarkably decreased mechanical allodynia. Fexofenadine (10 and 30 mg/kg, p.o) significantly attenuated the mechanical allodynia and prevented cold plate avoidance. Ranitidine (5 and 15 mg/kg, i.p) significantly prevented cold plate avoidance behavior and at the dose of 15 mg/kg notably improved mechanical and cold allodynia. Famotidine (1 and 3 mg/kg, p.o) was ineffective on all nociceptive tests. Gabapantin (100 mg/kg, p.o) significantly improved all types of nociceptive behaviors. These results indicate that both blood brain barrier penetrating (chlorpheniramine) and poorly penetrating (fexofenadine) histamine H 1 receptor antagonists could improve the neuropathic pain sign, but only the blood brain barrier penetrating histamine H 2 receptor antagonist (ranitidine) could produce anti-allodynic effects in the TNT model of neuropathic pain in rats. [ABSTRACT FROM AUTHOR]

Drugs of abuse trigger a very specific type of memory called state-dependent memory (SDM). Both memory process and drug addiction are underlain by neuroplasticity, which depends on calcium concentration and protein kinase activity. Within the scope of this study was to evaluate the impact of verapamil, an L-type voltage-gated calcium channel (VGCC) blocker, and SL-327, a selective MAPK/ERK kinase inhibitor, on morphine and ethanol SDM including the cross effects between these drugs with an additional influence of nicotine. To assess SDM in mice a step-through passive avoidance task was used. Our results show that amnestic effects of morphine (10.0 mg/kg, s.c.) and ethanol (1.0 g/kg, i.p.) can be reversed by pre-test administrations of morphine (10.0 mg/kg, s.c.), ethanol (1.0 g/kg, i.p.) and nicotine (0.1 mg/kg, s.c.), indicating morphine and ethanol SDM, as well as morphine-ethanol, morphine-nicotine, ethanol-morphine and ethanol-nicotine cross SDM. Pre-test co-treatment of verapamil (10.0 mg/kg, i.p.) with morphine/ethanol/nicotine increased all investigated SDM and cross SDM effects. Pre-test co-treatment of SL-327 (10.0 mg/kg, i.p.) diminished morphine- and ethanol-induced SDM along with the cross effects except ethanol-morphine cross SDM. In conclusion, SDM depends on ERK1/2 activation and also verapamil affects this type of memory, although the exact mechanism of its cognitive action has not been investigated in this study. [ABSTRACT FROM AUTHOR]

Acute coronary arterial spasm is contributory to ethanol-induced heart ischemic events. The present experiments were designed to study contractive effect of ethanol in isolated rat coronary arteries (RCAs) stimulated mildly with vasoconstrictors and involvement of voltage-gated potassium (K V ) channels and mitogen-activated protein kinases (MAPKs) of rat coronary arterial smooth muscle cell (RCASMC) in the spasm. The vascular tension was recorded with a wire myograph. K V currents of single freshly isolated RCASMC were assessed with patch clamp. Phosphorylation of RCASMC MAPKs (p38 MAPK and p44/42 MAPK) was assayed by Western blots. Mild stimulation, which was defined as 5–20% of 60 mM KCl-induced contraction, with thromboxane A 2 mimetic U46619, endothelin-1 or KCl tremendously increased contractive response of RCAs to ethanol (0.8–8.0 mg/ml). Ethanol (8 and 25 mg/ml) reduced the maximal K V currents by 53.6% and 56.6% respectively without concentration-dependence. Both ethanol (8.0 mg/ml) and U46619 (0.3 μM) enhanced phosphorylation of p38 MAPK and p44/42 MAPK and there was a pronounced synergism between them. MAPK pathway inhibition with p38 MAPK inhibitor SB239063 and p44/42 MAPK inhibitor PD98059 depressed ethanol-induced contraction by 22–55% in the stimulated RCAs and almost abrogated ethanol-induced reduction in K V currents. The present results for the first time demonstrated that ethanol induces potent vasoconstriction in stimulated RCAs and depresses K V currents in RCASMCs. It is suggested that MAPKs pathways play an important role in the etiology of ethanol-induced coronary artery diseases. [ABSTRACT FROM AUTHOR]

Marrow proliferation of the ossicular chain is a rare phenomenon. To date, only two other cases have described this rarity. We report a third paediatric case from Australia. A seven-year-old with thalassemia major demonstrated conductive impairment during surveillance for Deferasirox ototoxicity. Otitis media was assumed, however, CT scan of the petrous temporal bone revealed extramedullary haematopoiesis causing bilateral ossicular expansions and fixed conductive deficit. Reports of hearing loss in the thalassemia population focus on sensorineural impairment from iron chelation therapies. Clinicians should suspect ossicular deformation where treatment has been delayed, poorly controlled or conductive deficit persists without effusion. [ABSTRACT FROM AUTHOR]

Introduction Cochlear nerve aplasia or hypoplasia is found in up to a half of patients with unilateral or bilateral hearing loss. There is an ongoing discussion regarding the indication of cochlear implants for hearing rehabilitation in cases with radiologically-defined aplasia or hypoplasia of the cochlear nerve in those patients, especially in children. At present there is conflicting evidence whether the audiological outcomes of those children with a CI are comparable to those of children with a CI and a radiologically-normal cochlear nerve. The primary aim of this study was to assess the audiological abilities before and after CI provision in children with cochlear nerve hypoplasia or aplasia. Additionally, we aimed to determine if audiological outcomes differed in children with aplasia from those with hypoplasia. Such data should be helpful in determining if CI provision is appropriate for such children. Methods This retrospective study presents 7 children who were diagnosed with cochlear nerve aplasia or hypoplasia and received a CI. The pre- and postoperative audiological performance and the hearing and speech development of the children were examined. Results 4 children were unilateral CI users and 3 were bilateral CI users. Hearing reactions could be detected in all children. Already at first fitting, prompt responses and reactions to songs were observed. The aided thresholds in free field in children with hypoplasia were between 30 and 60 dB. Even in children with aplasia, the results in free field with CI averaged between 30 and 70 dB. Therefore the aided thresholds in children with hypoplasia and in children with aplasia of the CN are similar. It could be demonstrated that hearing reactions improve with the long term use of the implant. Improvement in general development could be observed in all children despite the very heterogeneous conditions and the accompanying handicaps. Conclusion The results of this study support the hypothesis that children with radiologically-defined CN hypoplasia or aplasia and detectable responses to electrical or acoustical stimuli can improve their sound detection thresholds and their awareness of sound when provided with a CI. [ABSTRACT FROM AUTHOR]

Objective Acute rhinosinusitis (ARS) is among the most common infectious diseases. Neutrophils play a major role in innate host defenses against pathogenic microorganisms such as fungi and bacteria. Recently, in neutrophils, ligation of the triggering receptor expressed on myeloid cells (TREM)-1 was found to activate the full spectrum of neutrophil effector mechanisms, including the release of inflammatory mediators, degranulation, phagocytosis, and oxidative burst in synergy with Toll-like receptors (TLRs). In this study, we investigated the effect of TREM-1 on the functions of neutrophils in relation to TLR4 in a nasal and nasopharyngeal inflammation mouse model via nontypeable Haemophilus influenzae (NTHi) intranasal inoculation. Methods We used C3H/HeJ (TLR4-deficient) mice, which arose spontaneously and have non-functional TLR4 protein, and normal wild-type (WT) C3H/HeN mice. Mice were inoculated intranasally with NTHi (10 7  cfu/mouse) to investigate the effects of TLR4 on the function of Neutrophils. We examined the kinetics of bacterial clearance and inflammatory cell infiltration in nasal washes at 6, 12, 24, and 72 h after inoculation. The expression of TREM-1 on neutrophils, and TREM-1 mRNA expression in neutrophils in the nasal washes were examined by flow cytometric analysis and RT-PCR. Results Bacterial counts of NTHi from nasal washes were significantly lower in WT mice than in TLR4-mutant mice after inoculation. The numbers of inflammatory cells in nasal washes were significantly higher in WT mice at 6 h, 12 h, and 24 h after inoculation than in TLR4-deficient mice. The expression of TREM-1 protein on neutrophils and the mRNA levels were greater in WT mice than in TLR4-mutant mice. The concentrations of soluble TREM-1 in WT nasal washes were also significantly higher than in those of TLR4-deficient mice. Conclusion TREM-1 may play an important role together with TLR4 in the nasopharyngeal clearance of NTHi by neutrophils. Further studies will need to clarify the innate immune responses of neutrophils via TLR4 to prevent NTHi infection. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) on the estrous response, ovulatory follicle development, ovulation rate, corpus luteum (CL) diameter, concentrations of progesterone (P4) and pregnancy per AI (P/AI) in CIDR-EB treated lactating anovular Nili-Ravi buffalo during the breeding season. Buffalo ( n = 87), 60 to 200 Days postpartum were treated with CIDR at a random stage of the estrous cycle (Day 0) and assigned to two groups; with eCG (+eCG; n  = 44), or without eCG (-eCG; n  = 43). In both groups, each animal received 2 and 1 mg estradiol benzoate (EB) on Day 0 and then on Day 8, respectively. On Day 6, eCG and PGF 2α were administered. On Day 7, CIDR were removed and FTAI was performed at 48 and 60 h later and pregnancy was diagnosed 35 Days post AI. The results revealed that eCG treatment increased ( P  <  0.05) the CL diameter on 15, 18, and 21 after the ovulation that resulted from imposing the estrous synchronization protocol than when eCG was not administered. Similarly, treatment with eCG resulted in an increased ( P  <  0.05) mean concentration of P4 as compared with animals not treated with eCG on Days 18 and 21 after the ovulation that occurred subsequent to imposing the estrous synchronization protocol. The ovulation rate was greater in eCG-treated buffalo as compared with those not treated with eCG (93% compared with 74%, respectively; P < 0.05). The P/AI was greater in buffalo treated with eCG than those not treated with eCG (56% compared with 31%, respectively; P < 0.05). In conclusion, the addition of eCG along with EB and P4 based treatments enhanced the estrous response, follicular and luteal function, ovulation rate, concentrations of P4, and P/AI in lactating anovular Nili-Ravi buffalo. [ABSTRACT FROM AUTHOR]

Sudden death syndrome (SDS) of soybean can be caused by at least four distinct Fusarium species, with F. tucumaniae being the main causal agent in Argentina. The fungus is a soil-borne pathogen that is largely confined to the roots, but damage also reaches aerial part of the plant and interveinal chlorosis and necrosis, followed by premature defoliation can be observed. In this study, two genetically diverse soybean cultivars, one susceptible (NA 4613) and one partially resistant (DM 4670) to SDS infection, were inoculated with F. tucumaniae or kept uninoculated. Leaf samples at 7, 10, 14 and 25 days post-inoculation (dpi) were chosen for analysis. With the aim of detecting early markers that could potentially discriminate the cultivar response to SDS, gas chromatography–mass spectrometry (GC–MS) analyses and biochemical studies were performed. Metabolic analyses show higher levels of several amino acids in the inoculated than in the uninoculated susceptible cultivar starting at 10 dpi. Biochemical studies indicate that pigment contents and Rubisco level were reduced while class III peroxidase activity was increased in the inoculated susceptible plant at 10 dpi. Taken together, our results indicate that the pathogen induced an accumulation of amino acids, a decrease of the photosynthetic activity, and an increase of plant-specific peroxidase activity in the susceptible cultivar before differences of visible foliar symptoms between genotypes could be observed, thus suggesting that metabolic and biochemical approaches may contribute to a rapid characterization of the cultivar response to SDS. [ABSTRACT FROM AUTHOR]

Ketamine and phencyclidine are well-known drugs of abuse of the arylcyclohexylamine class, the backbone of which is used for the synthesis of new psychoactive substances (NPS). In October 2017, a cluster of acute intoxications was encountered where patients presented with ketamine-like toxidrome. Upon initial toxicology screening, however, neither ketamine nor other causative agents were detected in the patients' urine. Instead, an unidentified substance was consistently detected. Further investigations using gas- and liquid-chromatography mass spectrometry led to the identification of an arylcyclohexylamine analogue, 2-oxo-PCE. The present study reports the analytical and toxicological profile of this emerging NPS. Chart review found, in total, 56 cases of 2-oxo-PCE associated acute poisoning between October and November 2017. Laboratory analysis confirmed the presence of 2-oxo-PCE in the urine of all patients; nasal swab samples from three patients revealed the lone presence of 2-oxo-PCE. Urine bedside immunoassay for ketamine was found not to cross-react with 2-oxo-PCE. In 55% of the cases, other drugs of abuse were detected on toxicology analysis; whilst in the remainder, 2-oxo-PCE was used alone. The main clinical symptoms associated with sole 2-oxo-PCE use include impaired consciousness (84%), confusion (60%), abnormal behaviour (44%), hypertension (80%) and tachycardia (40%). Convulsion (16%) was also observed relatively frequently. Management was mainly supportive, whilst three patients required intensive care. All patients recovered uneventfully. In conclusion, frontline clinical and laboratory personnel should be highly vigilant in the lookout for 2-oxo-PCE, a dangerous emerging arylcyclohexylamine analogue. [ABSTRACT FROM AUTHOR]

Although leukocyte telomere length (TL) shortens over the lifespan and is associated with diseases of aging, little is known about the relationships between TL, memory, and brain structure. Sixty-nine functionally normal older adults (mean age = 71.7) were assessed at 2 time points (mean interval = 2.9 years). Linear mixed models assessed relationships between TL and hippocampal volume, fractional anisotropy, and mean diffusivity (MD) of the fornix and verbal and visual episodic memory. Unstandardized coefficients are reported in the following, and p values are not corrected for multiple comparisons. A negative baseline trend was observed between TL and fornix MD ( b = −0.01, p = 0.06), but no other cross-sectional associations were significant ( p s > 0.16). Greater TL shortening at follow-up was associated with greater hippocampal volume loss ( b = 27.09, p < 0.001), even after controlling for global volume loss ( b = 10.83, p = 0.002). Greater telomere attrition was also associated with larger increases in fornix MD ( b = −0.01, p = 0.012) and decreases in fornix fractional anisotropy ( b = 0.004, p = 0.002). TL was not associated with changes in episodic memory ( p s > 0.23). These relationships may reflect neurobiological influences that affect both TL and brain structure, as well as the effect of TL on brain aging via mechanisms such as cellular senescence and inflammation. [ABSTRACT FROM AUTHOR]

Background: Rapid detection of changes in cardiac index (CI) in real time using minimally invasive monitors may be of clinical benefit. We tested whether the Starling-SV bioreactance device, which averages CI over a short 8 s period, could assess the effects of passive leg raising (PLR), a clinical test that is recommended to assess fluid responsiveness during septic shock.Methods: In 32 critically ill patients, we measured CI by transpulmonary thermodilution (PiCCO2, CItd), pulse contour analysis (PiCCO2, CIPulse), and the Starling-SV device (CIStarling) at baseline. CIPulse and CIStarling were measured again at the end of a PLR test. In the 13 patients with a positive PLR test, CItd, CIPulse, and CIStarling were measured before and after a 500 ml saline infusion. The primary outcome was relative changes from baseline measurements in CItd, CIPulse, and CIStarling. Secondary outcomes compared absolute values measured by each method.Results: Relative changes in CIPulse and CItd were significantly correlated (r=0.82; n=45; P<0.001), with an 89% concordance rate (n=45 paired measurements). Relative changes in CIStarling and CItd were also significantly correlated (r=0.59; n=45; P<0.001) with a 78% concordance rate. For absolute measures of CI (n=77 paired measurements), the bias between CIPulse and CItd was 0.01 L min-1 m-2 (limits of agreement, -0.49 and 0.51 L min-1 m-2; 15% percentage error). Bias between CIStarling and CItd was 0.03 L min-1 m-2 (limits of agreement, -1.61 and 1.67 L min-1 m-2; 48% percentage error).Conclusions: In critically ill patients, a non-invasive bioreactance device with a shorter averaging period assessed a passive leg raising test with reasonable accuracy. [ABSTRACT FROM AUTHOR]

Small collagen peptides are associated with various benefits, such as bone and skin health. However, preparation of small collagen peptides from terrestrial vertebrate origins remains a challenge. Here, we show that pretreatment with formic acid enhanced the production of small collagen peptides from spent hen skin. After pretreatment with formic acid, the percentage of small peptides below 2 kDa increased to 48.92% and 43.34% from 33.79% and 36.32% for heat-soluble collagen (HSC) and pepsin-soluble collagen (PSC), respectively. Pretreatment with formic acid degraded telopeptides and released the cross-links (pyrrole and pyridinoline), which made hen collagen more susceptible to papain hydrolysis. LC-MS/MS results revealed that none of the peptides identified from HSC-FA (formic acid)-Papain and PSC-FA-Papain were derived from cross-linked telopeptides. These results demonstrated that formic acid assisted the hydrolysis of highly cross-linked collagen of spent hens, and it might also be used to produce small collagen peptides from other aged, vertebrate collagens. [ABSTRACT FROM AUTHOR]

Oxidative stress and blood-brain barrier (BBB) dysfunction contribute to brain injury after intracerebral hemorrhage (ICH). Adiponectin (APN) inhibits oxidative stress in the CNS, but the role of APN in ICH is not clear. Thus, we elucidated the possible neuroprotective effect of APN in ICH-induced brain injury in rats and investigated the neuroprotective mechanisms. A lentivirus-carrying APN gene was injected into rats 14 days before ICH induced via intracerebral injection of autologous blood. The effects of lentiviral overexpression of APN on brain injury were evaluated 24 h after ICH. Superoxide dismutase (SOD), glutathione (GSH), and the ratio of oxidized glutathione to reduced glutathione (GSSG/GSH) and malondialdehyde (MDA) were measured. Oxidative stress-related proteins were measured by Western blot and qRT-PCR. APN overexpression improved neurological function, reduced brain edema, preserved the BBB and increased the expression of APN and decreased the expression of NADPH oxidase-2 (NOX 2) compared with null vector controls ( p <  0.01). SOD, GSH, and GSSG/GSH increased, and MDA was reduced. Furthermore, tetrabromocinnamic acid (TBCA, a NADPH oxidase activator) blocked the effect of APN on cerebral protection and antioxidant activity. Our results demonstrate the importance of APN in regulating oxidative stress and BBB function and suggest APN may be a novel treatment for brain damage after ICH. [ABSTRACT FROM AUTHOR]

Background: Inulin-type fructans used in formula have been shown to promote microbiota composition and stool consistency closer to those of breastfed infants and to have beneficial effects on fever occurrence, diarrhea, and incidence of infections requiring antibiotic treatment in infants.Objectives: The primary study aim was to explore whether prophylactic supplementation with prebiotic fructans is able to influence the frequency of infectious diseases in kindergarten children during a winter period. A secondary objective was to ascertain the effect on the intestinal microbiota.Methods: 142 boys and 128 girls aged 3-6 y were randomly allocated to consume 6 g/d fructans or maltodextrin for 24 wk. At baseline, stool samples were collected for microbiota analysis and anthropometric measurements were made. During the intervention period diagnoses were recorded by physicians, whereas disease symptoms, kindergarten absenteeism, dietary habits, and stool consistency were recorded by parents. Baseline measurements were repeated at wk 24.Results: In total 219 children finished the study. Both the relative abundance of Bifidobacterium (P < 0.001) and that of Lactobacillus (P = 0.014) were 19.9% and 7.8% higher, respectively, post data normalization, in stool samples of children receiving fructans as compared with those of controls at wk 24. This was accompanied by significantly softer stools within the normal range in the prebiotic group from wk 12 onwards. The incidence of febrile episodes requiring medical attention [0.65 ± 1.09 compared with 0.9 ± 1.11 infections/(24 wk × child), P = 0.04] and that of sinusitis (0.01 ± 0.1 compared with 0.06 ± 0.25, P = 0.03) were significantly lower in the prebiotic group. The number of infectious episodes and their duration reported by parents did not differ significantly between the 2 intervention groups.Conclusions: Prebiotic supplementation modified the composition of the intestinal microbiota and resulted in softer stools in kindergarten-aged children. The reduction in febrile episodes requiring medical attention supports the concept of further studies on prebiotics in young children. This trial was registered at clinicaltrials.gov as NCT03241355. [ABSTRACT FROM AUTHOR]

Background: Zearalenone (ZEN) can cause serious defects in development and reproduction in humans and animals. Silymarin shows antioxidant and estrogenic effects.Objective: This study was conducted to determine if silymarin can antagonize ZEN-induced hepatic and reproductive toxicities.Methods: Thirty-five 21-d-old female Sprague-Dawley rats (n = 7/diet) were fed a control diet (Ctrl) or Ctrl plus 20 mg ZEN/kg or Ctrl plus 20 mg ZEN/kg with 100, 200, or 500 mg silymarin/kg for 6 wk. Serum, livers, ovaries, and uterus were collected at week 6 for biochemistry, hormone, and redox status and selected gene and protein assays.Results: The consumption of ZEN decreased (P < 0.05) the final body weight by 17.9%, induced liver injury, increased (P < 0.05) aspartate aminotransferase and alkaline phosphatase activities, and decreased (P < 0.05) total protein and albumin concentrations in serum by 16.7-40.6%. ZEN also caused reproductive toxicity, including decreased (P < 0.05) 17β-estradiol and increased (P < 0.05) follicle-stimulating hormone concentrations in serum by 12.7-46.3% and induced histopathologic alterations in the liver, ovaries, and uterus. Interestingly, these alterations induced by ZEN were alleviated (P < 0.05) by silymarin supplementation at 100, 200, and 500 mg/kg. Moreover, silymarin supplementation at the 3 doses mitigated (P < 0.05) ZEN-induced impairment in hepatic glutathione peroxidase activity, total antioxidant capacity, and malondialdehyde concentration by 17.6-100%. Meanwhile, silymarin supplementation at all doses upregulated (P < 0.05) phospho-ribosomal protein S6 kinase 1 (p-RPS6KB1) and 3β-hydroxysteroid dehydrogenase (HSD3B) by 43.0-121% but downregulated (P < 0.05) AMP-activated protein kinase (AMPK) and 3α-hydroxysteroid dehydrogenase (HSD3A) in the liver relative to the ZEN group by 11.2-40.6%. In addition, silymarin supplementation at all doses elevated (P < 0.05) HSD3B by 1.8- to 2.5-fold and decreased (P < 0.05) estrogen receptor 1 (ESR1), ATP binding cassette (ABC) c1, and Abcc5 in ovaries and the uterus by 10.7-63.2%.Conclusion: Dietary silymarin supplementation at 100, 200, and 500 mg/kg protected rats from ZEN-induced hepatotoxicity and reproductive toxicity, potentially through improvement in the antioxidant capacity and regulation in the genes related to protein synthesis, ZEN metabolism, hormone synthesis, and ABC transporters in the tissues. [ABSTRACT FROM AUTHOR]

Background: Iron deficiency is an enduring global health problem that requires new remedial approaches. Iron absorption from soybean-derived ferritin, an ∼550-kDa iron storage protein, is comparable to bioavailable ferrous sulfate (FeSO4). However, the absorption of ferritin is reported to involve an endocytic mechanism, independent of divalent metal ion transporter 1 (DMT-1), the transporter for nonheme iron.Objective: Our overall aim was to examine the potential of purified ferritin from peas (Pisum sativum) as a food supplement by measuring its stability under gastric pH treatment and the mechanisms of iron uptake into Caco-2 cells.Methods: Caco-2 cells were treated with native or gastric pH-treated pea ferritin in combination with dietary modulators of nonheme iron uptake, small interfering RNA targeting DMT-1, or chemical inhibitors of endocytosis. Cellular ferritin formation, a surrogate measure of iron uptake, and internalization of pea ferritin with the use of specific antibodies were measured. The production of reactive oxygen species (ROS) in response to equimolar concentrations of native pea ferritin and FeSO4 was also compared.Results: Pea ferritin exposed to gastric pH treatment was degraded, and the released iron was transported into Caco-2 cells by DMT-1. Inhibitors of DMT-1 and nonheme iron absorption reduced iron uptake by 26-40%. Conversely, in the absence of gastric pH treatment, the iron uptake of native pea ferritin was unaffected by inhibitors of nonheme iron absorption, and the protein was observed to be internalized in Caco-2 cells. Chlorpromazine (clathrin-mediated endocytosis inhibitor) reduced the native pea ferritin content within cells by ∼30%, which confirmed that the native pea ferritin was transported into cells via a clathrin-mediated endocytic pathway. In addition, 60% less ROS production resulted from native pea ferritin in comparison to FeSO4.Conclusion: With consideration that nonheme dietary inhibitors display no effect on iron uptake and the low oxidative potential relative to FeSO4, intact pea ferritin appears to be a promising iron supplement. [ABSTRACT FROM AUTHOR]