Increase In Inflammatory Cells Research Articles

A Series of Compounds Bearing a Dipyrido-Pyrimidine Scaffold Acting as Novel Human and Insect Pest Chitinase Inhibitors.

Chitinases not only play vital roles in the human innate immune system but are also essential for the development of pathogenic fungi and pests. Chitinase inhibitors are efficient tools to investigate the elusive role of human chitinases and to control pathogens and pests. Via hierarchical virtual screening, we have discovered a series of chitinase inhibitors with a novel scaffold that have high inhibitory activities and selectivities against human and insect chitinases. The most potent human chitotriosidase inhibitor, compound 40, exhibited a Ki of 49 nM, and the most potent inhibitor of the insect pest chitinase OfChi-h, compound 53, exhibited a Ki of 9 nM. The binding of these two most potent inhibitors was confirmed by X-ray crystallography. In a murine model of bleomycin-induced pulmonary fibrosis, compound 40 was found to suppress the chitotriosidase activity by 60%, leading to a significant increase in inflammatory cells and suggesting that chitotriosidase played a protective role.

Juglone Attenuates Sepsis-Induced Acute Lung Injury via Suppression of the TLR4/Nf-κb Pathway

Sepsis, with high mortality, induces deleterious organ dysfunction and acute lung injury. Natural compounds show protective effect against sepsis-induced acute lung injury. Juglone, a natural naphthoquinone, demonstrates pharmacological actions as a pro-apoptotic substrate in tumor treatment and anti-inflammation substrate in organ injury. In this study, the influence of juglone on sepsis-induced acute lung injury was investigated. First, a septic mice model was established via cecal ligation and puncture, and then verified via histopathological analysis of lung tissues, the wet/dry mass ratio and myeloperoxidase activity was determined. Cecal ligation and puncture could induce acute lung injury in septic mice, as demonstrated by alveolar damage and increase of wet/dry mass ratio and myeloperoxidase activity. However, intragastric administration juglone attenuated cecal ligation and puncture-induced acute lung injury. Secondly, cecal ligation and puncture-induced increase of inflammatory cells in bronchoalveolar lavage fluid was also alleviated by the administration of juglone. Similarly, the protective effect of juglone against cecal ligation and puncture-induced acute lung injury was accompanied by a reduction of pro-inflammatory factor secretion in bronchoalveolar lavage fluid and lung tissues. Cecal ligation and puncture could activate toll-like receptor 4/nuclear factor-kappa B signaling pathway, and administration of juglone suppressed toll-like receptor 4/nuclear factor-kappa B activation. In conclusion, juglone attenuated cecal ligation and puncture-induced lung damage and inflammatory response through inactivation of toll-like receptor 4/nuclear factor-kappa B, suggesting a potential therapeutic strategy in the treatment of sepsis-induced acute lung injury.

Oxidative stress and immune complexes: Pathogenic mechanisms in pristane induced murine model of lupus.

Systemic lupus erythematosus is a systemic autoimmune inflammatory disease with a broad spectrum of clinical presentations. Mouse models play an indispensible role in understanding the disease pathology and for developing new therapeutics. This study investigated the effect of pristane administration on female BALB/c mice. The various manifestations of lupus replicated in this mouse model were analysed and their relevance to human disease was assessed. Pristane injection replicates the two prime manifestations of lupus i.e. oxidative stress and inflammation. An increase in oxidative stress was determined by the decreasing anti-oxidants, increasing ROS and lipid peroxidation. Inflammatory cytokines (IL-6 and TNF-α) also increased in the plasma. The deteriorating organ functions after pristane administration were evident histopathologically. An increase in inflammatory cells, necrosis, oil vacuoles and pigment cells were marked in kidney, liver, lungs and spleen, whereas skin did not manifest any alteration. Liver function (bilirubin, SGPT) and kidney function (creatinine and proteinuria) tests were also altered. Pristane injection caused the generation of anti-nuclear antibodies, which were apparent from the different immunofluorescence patterns observed. Immune deposits were evident in all the vital organs stating the similarity this model holds with lupus patients. Replicating various manifestations of human lupus disorder, pristane induced mouse model of lupus serves as an appropriate model to study lupus complications and in the development of novel therapeutics for disease management.

Thymic stromal lymphopoietin (TSLP) and Toluene-diisocyanate-induced airway inflammation: Alleviation by TSLP neutralizing antibody

Toluene-diisocyanate (TDI) is mainly used in the manufacturing process of polyurethane foams, and is a potent inducer of occupational asthma characterized by airway inflammation and airway hyperreactivity. Thymic stromal lymphopoietin (TSLP) plays an important role in the development of asthma, and correlating with the differentiation of Th2 and Th17 cells. However, the role of TSLP in TDI-induced asthma remains unclear. In this study, 96 TDI-exposed workers as well as a mouse model of TDI-induced asthma were investigated. The air exposure assessment result of TDI in the workplace showed that workers were exposed to inhalation of a very high concentration of TDI, approximately 8 times the recommended level, leading to a decrease in pulmonary function and an increase in inflammatory cells, as well as TSLP and IgE levels in the supernatant of sputum obtained from exposed workers. In order to further investigate the role of TSLP in the pathogenesis of TDI-induced asthma, a mouse model of TDI-induced asthma was also employed. Histopathological analysis of mouse lung and bronchus showed an obvious infiltration of inflammatory cells around the bronchus. The levels of inflammatory cells, IFN-γ, IL-4 and IL-17 in bronchoalveolar lavage fluid (BALF), the expression levels of TSLP protein and ROR-γt and IL-17 mRNA in mouse lung tissues were also significantly increased. However, after treatment with TSLP neutralizing antibody (TSLP-Ab), the degree of pulmonary and bronchial inflammation in mice was significantly alleviated, and the levels of inflammatory cells, IFN-γ, IL-4 and IL-17 in BALF, and the expression levels of ROR-γt and IL-17 mRNA in lung tissue were significantly decreased. Our data shows that TSLP plays an important role in the pathogenesis of TDI-induced asthma, and that TSLP-Ab can effectively alleviate TDI-induced airway inflammation of asthma.

Suppression of NLRP3 and NF-κB signaling pathways by α-Cyperone via activating SIRT1 contributes to attenuation of LPS-induced acute lung injury in mice

α-Cyperone is the volatile oil component in Cyperus rotundus L. The previous reports had shown the inhibition of α-Cyperone on NF-κB signaling in LPS-stimulated RAW264.7 cells. However, it is still unclear whether α-Cyperone could suppress inflammatory response in acute lung injury (ALI) induced by LPS. The aim of this study is to investigate the suppression of α-Cyperone on LPS-induced ALI in mice. In this study, we established the LPS-induced ALI model and compared different doses of α-Cyperone with the control group and LPS group. Accordingly, the following indexes would be compared, including lung wet/dry ratio, MPO activity, inflammatory cell number, histopathological changes, levels of inflammatory cytokines, NF-κB and NLRP3 signaling pathways activation. The results demonstrated that α-Cyperone had the effect on reducing the wet/dry ratio and MPO activity. Furthermore, the increase of inflammatory cells and inflammatory cytokines could be inhibited by α-Cyperone. Meanwhile, α-Cyperone could downregulate NF-κB and NLRP3 signaling pathways. Finally, we found α-Cyperone could up-regulate the expression of SIRT1 and SIRT1 inhibitor could reverse the protective effects of α-Cyperone on ALI. In conclusion, α-Cyperone showed the protective effect on LPS-induced ALI in mice by suppressing the NF-κB and NLRP3 signaling pathways, mainly via up-regulating SIRT1. This provides a potential drug for the treatment of LPS-induced ALI.

Abstract 3760: Identifying site-specific disparities in the tumor microenvironment of primary and metastatic disease

Abstract The tumor microenvironment (TME), or all cellular and molecular components in and around the tumor, has a well-documented impact on cancer cell proliferation, chemoresistance, and overall patient survival. However much of the research into the TME has been limited to the primary site. How the TME of metastases compares to the primary lesions remains poorly characterized, even though metastasis is responsible for 90% of all cancer deaths. Understanding these differences is critical to treating advanced cancer patients. To study this our lab examined the TME in 22 matched sets primary pancreatic tumors and liver metastasis obtained from end-stage pancreatic cancer patients via our rapid autopsy program. We used a multiplex-immunofluorescent technique that allows staining of a single slide with up to 35 antigens and characterized the presence, position and functional statues of cells that comprise the pancreatic TME. These included: tumor cells, vasculature, fibroblasts, macrophages, mast cells, basophils, eosinophils, B cells, T cells, and NK cells, along with multiple subtypes of these. Based on our preliminary data from this, we hypothesize that the organ determines the features of the TME. Analyzing primary tumors and metastases revealed that both the composition and spatial distribution of different cell types are distinct between the primary tumor and liver metastases. PAM clustering of the cellular featured grouped primary tumor with primary and metastasis with metastatis 87% of the time. As the TME has been shown to impact chemotherapeutic response, we further examined chemotherapeutic effect between sites. Comparing gemcitabine (standard of care) and untreated patients, we observed distinct site-based responses. In the primary tumors with gemcitabine treatment there was a significant increase in inflammatory cells (neutrophils and macrophages) in comparison to untreated patients, while in the liver gemcitabine induced an increase in desmoplasia (SMA). Combined this data indicates that the distinct TME’s exist within the same patient in an organ specific manner which can impact chemotherapeutic response. Understand these differences is therapeutically critical to patients whose therapeutic strategies involve targeting the TME to improve drug delivery, modulate immune suppression or otherwise modulate the microenvironment. Citation Format: Krysten E. Vance, Micheal A. Hollingsworth, Paul Grandgenett. Identifying site-specific disparities in the tumor microenvironment of primary and metastatic disease [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3760.

DIFFERENTIAL EXPRESSION OF PD1 AND PDL1 IN ORAL POTENTIALLY MALIGNANT LESIONS AND ORAL SQUAMOUS CELL CARCINOMA:A PILOT STUDY

Background Programmed cell death protein 1 (PD-1, CD279) is a 50-55 kDa type I transmembrane receptor expressed by activated T and B cells, as well as subset of monocytes and dendritic cells (DCs). PD-1 and its ligands (PDL1, PDL2) are part of “checkpoint” immune recognition and peripheral tolerance system that emerged as a critical signaling pathway in cancer. PDL1 is expressed in various types of cancers and activation of PD1-PDL1 inhibits T-cell mediated cancer surveillance. Here we describe a quantitative, reproducible 2-color fluorescence-based protocol to determine the differential expression of PD1/PDL1 in oral biopsy specimens. Methods Histopathological samples with a diagnosis of hyperkeratosis (HK), OMPL (mild, moderate, severe dysplasia) and squamous cell carcinoma (OSCC) were selected from the archives of the Toronto Oral Pathology service, University of Toronto. FFPE sections were stained with monoclonal antibodies for PD1 and PDL1 (Abcam) and Alexa Fluor-labelled secondary antibodies allowing visualization of both proteins in the same section using a spinning disk confocal microscope (Quorum). PDL1 staining was assessed in basal/spinous layers of the epithelium while PD1 staining was assessed in inflammatory cells in tumor stroma/lamina propria. The mean fluorescent intensity (MFI) was quantified and normalized against background signal. Results Our results show a significant increase in PD1 expression in inflammatory cells in dysplasia and OSCC compared to hyperkeratosis. PDL1 expression in epithelial cells was significantly increased in OSCC but not in dysplasia or HK. The results suggest that PD1 increase in inflammatory cells precedes malignant transformation while PDL1 overexpression in epithelial cells only occurs after malignant transformation. Conclusion We developed a new quantitative method to study PD1/PDL1 expression in FFPE oral biopsy samples. The expression of PD1 and PDL1 may be used as predictive markers of transformation and the data may be used to develop early intervention in OPML using PD1 inhibitors.

Ccf-mtDNA as a Potential Link Between the Brain and Immune System in Neuro-Immunological Disorders.

Fragments of mitochondrial DNA (mtDNA) are released outside the cell and they appear to persist in extracellular fluids as circulating, cell-free, mtDNA (ccf-mtDNA). When compared to nuclear DNA, such a double stranded mtDNA is more resistant to nuclease degradation. In fact, it is stable extracellularly where it can be detected in both plasma and cerebrospinal fluid (CSF), here acting as a potential biomarker in various disorders. In neurological diseases (Alzheimer's disease, Parkinson's disease and end-stage progressive Multiple Sclerosis), a decreased amount of CSF ccf-mtDNA is related with progressive cell dysfunction. This suggests an alteration in neuronal mtDNA levels (mtDNA replication, degradation and depletion) in vulnerable brain regions at early stages of neurodegeneration leading to reduced mtDNA release, which takes place before actual cell death occurs. On the other hand, elevated CSF ccf-mtDNA levels are reported in acute phases of relapsing-remitting Multiple Sclerosis (RRMS). This occurs during acute inflammation, which anticipates the neurodegenerative process. Thus, an increase in inflammatory cells in the affected regions is expected to add on mtDNA release into the CSF. In addition, similarly to bacterial DNA, the non-methylated CpG sites of mtDNA, which activate innate immunity and inflammation, are likely to participate in the molecular mechanisms of disease. Thus, ccf-mtDNA may represent a powerful biomarker for disease screening and prognosis at early stage, although its biological role may extend to generating the neurobiology of disease. The present manuscript discusses recent experimental findings in relationship with clinical evidence comparing neuro-immunological features of neurodegenerative disorders with frankly neuro-infectious diseases.

Tumor necrosis factor alpha and keratin 17 expression in oral submucous fibrosis in rat model

Background and Objective: Oral submucous fibrosis is an inflammatory and a premalignant lesion as classified by “World Health Organization”. Its pathogenesis is related to collagen metabolism. Bleomycin is an anti-carcinogenic drug, but it causes pulmonary fibrosis. This study was carried out to study the role of tumor necrosis factor alpha and keratin 17 in pathogenesis of oral submucous fibrosis. Study Design: Fifty pathologically-free female rats were divided into one control group and four test groups. They were injected by phosphate-buffered saline and Bleomycin into the buccal mucosae daily for 2, 4, 6 and 8 weeks, respectively. The histopathological changes was evaluated by studying the changes within epithelium tissue and lamina propria by (H&E) and the expression of both (tumor necrosis factor alpha and keratin 17) using immunohistochemistry and enzyme linked immunosorbent assay respectively. Results: The buccal mucosae of the test groups in comparison with control showed hyperplastic and hyperatrophic epithelial changes that gradually decreased with the severity of oral submucous fibrosis, with epithelial atrophy and focal dysplastic changes, which increase with severity of oral submucous fibrosis gradually. The lamina propria showed gradual increase in inflammatory cells, collagen formation and closure of the blood vessels. Regarding tumor necrosis factor alpha the area percent of immunoexpression, showed the lowest expression in the control and test groups showed gradually increase after 2, 4, 6,8 weeks within epithelium , on the other hand, the area percent of immunoexpression of tumor necrosis factor alpha within lamina propria showed gradually increase after 2, 4, 6 weeks, then decreased after 8 weeks. Regarding Keratin-17, the lowest value was recorded in control group while it increased gradually from the second week to the 8th weeks.Conclusion: Both protein molecules tumor necrosis factor alpha and keratin 17 appear to be related in case of oral submucous fibrosis. As tumor necrosis factor alpha and keratin 17 are interdependent regulators, they could be used as diagnostic makers and a prognostic mirror of oral submucous fibrosis cases. Tumor necrosis factor alpha inhibitory drugs can be targeted in new therapeutic agents to improve the prognosis of oral submucous fibrosis.

Lung Protection Strategies during Cardiopulmonary Bypass Affect the Composition of Bronchoalveolar Fluid and Lung Tissue in Cardiac Surgery Patients.

Pulmonary dysfunction is among the most frequent complications to cardiac surgeries. Exposure of blood to the cardiopulmonary bypass (CPB) circuit with subsequent lung ischemia-reperfusion leads to the production of inflammatory mediators and increases in microvascular permeability. The study aimed to elucidate histological, cellular, and metabolite changes following two lung protective regimens during CPB with Histidine-Tryptophan-Ketoglutarate (HTK) enriched or warm oxygenated blood pulmonary perfusion compared to standard regimen with no pulmonary perfusion. A total of 90 patients undergoing CPB were randomized to receiving HTK, oxygenated blood or standard regimen. Of these, bronchoalveolar lavage fluid (BALF) and lung tissue biopsies were obtained before and after CPB from 47 and 25 patients, respectively. Histopathological scores, BALF cell counts and metabolite screening were assessed. Multivariate and univariate analyses were performed. Profound histological, cellular, and metabolic changes were identified in all patients after CPB. Histological and cellular changes were similar in the three groups; however, some metabolite profiles were different in the HTK patients. While all patients presented an increase in inflammatory cells, metabolic acidosis, protease activity and oxidative stress, HTK patients seemed to be protected against severe acidosis, excessive fatty acid oxidation, and inflammation during ischemia-reperfusion. Additional studies are needed to confirm these findings.

Rapid clearance of heavy chain-modified hyaluronan during resolving acute lung injury

BackgroundSeveral inflammatory lung diseases display abundant presence of hyaluronic acid (HA) bound to heavy chains (HC) of serum protein inter-alpha-inhibitor (IαI) in the extracellular matrix. The HC-HA modification is critical to neutrophil sequestration in liver sinusoids and to survival during experimental lipopolysaccharide (LPS)-induced sepsis. Therefore, the covalent HC-HA binding, which is exclusively mediated by tumor necrosis factor α (TNFα)-stimulated-gene-6 (TSG-6), may play an important role in the onset or the resolution of lung inflammation in acute lung injury (ALI) induced by respiratory infection.MethodsReversible ALI was induced by a single intratracheal instillation of LPS or Pseudomonas aeruginosa in mice and outcomes were studied for up to six days. We measured in the lung or the bronchoalveolar fluid HC-HA formation, HA immunostaining localization and roughness, HA fragment abundance, and markers of lung inflammation and lung injury. We also assessed TSG-6 secretion by TNFα- or LPS-stimulated human alveolar macrophages, lung fibroblast Wi38, and bronchial epithelial BEAS-2B cells.ResultsExtensive HC-modification of lung HA, localized predominantly in the peri-broncho-vascular extracellular matrix, was notable early during the onset of inflammation and was markedly decreased during its resolution. Whereas human alveolar macrophages secreted functional TSG-6 following both TNFα and LPS stimulation, fibroblasts and bronchial epithelial cells responded to only TNFα. Compared to wild type, TSG-6-KO mice, which lacked HC-modified HA, exhibited modest increases in inflammatory cells in the lung, but no significant differences in markers of lung inflammation or injury, including histopathological lung injury scores.ConclusionsRespiratory infection induces rapid HC modification of HA followed by fragmentation and clearance, with kinetics that parallel the onset and resolution phase of ALI, respectively. Alveolar macrophages may be an important source of pulmonary TSG-6 required for HA remodeling. The formation of HC-modified HA had a minor role in the onset, severity, or resolution of experimental reversible ALI induced by respiratory infection with gram-negative bacteria.

Repeated pulmonary exposures to zinc ions enhance inflammatory responses to subsequent metal exposures

Aim of Study: Metal contaminants contribute to adverse human health effects via acute and chronic exposures. Acute metal exposures followed by prolonged secondary metal exposures may elicit exaggerated inflammatory responses in certain individuals. The aim of this study is to determine whether repeated pulmonary exposures to zinc chloride (ZnCl2) alter subsequent responses to zinc or cerium exposures. Materials and Methods: Rats were intratracheally (IT) instilled with physiologic saline (n = 24) or 0.05 mg/kg ZnCl2 (n = 16) twice weekly for 4 weeks. Four days after last dosing, the saline group was divided into three subgroups, each IT-instilled with either saline, ZnCl2 or CeCl3 (both at 0.1 mg/kg). The ZnCl2 pre-instilled rats were divided into two subgroups, each instilled with 0.1 mg/kg ZnCl2 or CeCl3. Biomarkers of lung injury/inflammation were assessed in bronchoalveolar lavage (BAL) fluid collected 24 hours later. Oxidative stress was evaluated as total and reduced glutathione in BAL. Results: Increases in inflammatory cells, LDH, albumin, leptin, MCP-1, IP-10, fractalkine, TNFα and RANTES were observed in rats instilled with multiple PBS and then with 0.1 mg/kg ZnCl2 and CeCl3. However, rats pre-exposed repeatedly to 0.05 mg/kg ZnCl2 and then challenged with 0.1 mg/kg ZnCl2 or CeCl3 showed even more eosinophils, lymphocytes, and increased concentrations of hemoglobin and MIP-1α. Significant reduction in GSH/GSSG ratios in BAL in response to all ZnCl2 or CeCl3 exposures indicated oxidative stress. Conclusion: Previous exposure to zinc ions increases responsiveness to subsequent exposures to zinc and cerium ions. These findings suggest enhanced sensitization possibly due to a reduction in antioxidant defenses.

Progressive increase in allergen concentration abrogates immune tolerance in ovalbumin-induced murine model of chronic asthma

Persistent inflammation and remodeling of airways are the major hallmarks of asthma. Though airway inflammation diminishes in ovalbumin (OVA)-based mouse model of chronic asthma owing to immune-tolerance linked with repeated allergen exposure, which limits the application of the disease model. Accordingly, the present study was designed to develop a murine model of chronic asthma which presents persistent airway inflammation coupled with remodeling traits. Herein, OVA-sensitized BALB/c mice were challenged with increasing (modified protocol) or constant concentration (conventional protocol) of the allergen for 6 weeks; 3 times/week. The results, indeed, revealed that mice subjected to modified protocol demonstrate an improved response to the allergen as reflected by the significant increase in inflammatory cells particularly, eosinophils in bronchoalveolar lavage fluid compared to conventional protocol. Moreover, the expression of Th2 cytokines and their responsible transcription factors (GATA-3 and STAT-6) was markedly enhanced in lungs. The increase in inflammation was further accompanied by a marked increase in mucus production, collagen deposition, and the expression of allied factors (Muc5ac, Col1α1, and α-SMA). Interestingly, pre-treatment of dexamethasone, a corticosteroid (0.5 mg/kg b.wt., i.p.), suppressed the allergen-induced airway inflammation and mucus production without altering collagen deposition. Failure of dexamethasone seems to be related to their ineffectiveness to modulate the expression of TGF-β, MMP-9, COL1α1, and α-SMA. Overall, our results strongly suggest that mice underwent modified chronic protocol bears more resemblance with asthmatics as it imitates persistent airway inflammation allied with steroid-refractory remodeling traits; hence, may be useful for the evaluation of new/alternative drugs in steroid-refractory asthmatic conditions.

A novel surgical technique for a rat subcutaneous implantation of a tissue engineered scaffold

ObjectivesSubcutaneous implantations in small animal models are currently required for preclinical studies of acellular tissue to evaluate biocompatibility, including host recellularization and immunogenic reactivity.MethodsThree rat subcutaneous implantation methods were evaluated in six Sprague Dawley rats. An acellular xenograft made from porcine pericardium was used as the tissue-scaffold. Three implantation methods were performed; 1) Suture method is where a tissue-scaffold was implanted by suturing its border to the external oblique muscle, 2) Control method is where a tissue-scaffold was implanted without any suturing or support, 3) Frame method is where a tissue-scaffold was attached to a circular frame composed of polycaprolactone (PCL) biomaterial and placed subcutaneously. After 1 and 4 weeks, tissue-scaffolds were explanted and evaluated by hematoxylin and eosin (H&E), Masson’s trichrome, Picrosirius Red, transmission electron microscopy (TEM), immunohistochemistry, and mechanical testing.ResultsMacroscopically, tissue-scaffold degradation with the suture method and tissue-scaffold folding with the control method were observed after 4 weeks. In comparison, the frame method demonstrated intact tissue scaffolds after 4 weeks. H&E staining showed progressive cell repopulation over the course of the experiment in all groups with acute and chronic inflammation observed in suture and control methods throughout the duration of the study. Immunohistochemistry quantification of CD3, CD 31, CD 34, CD 163, and αSMA showed a statistically significant differences between the suture, control and frame methods (P < 0.05) at both time points. The average tensile strength was 4.03 ± 0.49, 7.45 ± 0.49 and 5.72 ± 1.34 (MPa) after 1 week and 0.55 ± 0.26, 0.12 ± 0.03 and 0.41 ± 0.32 (MPa) after 4 weeks in the suture, control, and frame methods; respectively. TEM analysis showed an increase in inflammatory cells in both suture and control methods following implantation.ConclusionRat subcutaneous implantation with the frame method was performed with success and ease. The surgical approach used for the frame technique was found to be the best methodology for in vivo evaluation of tissue engineered acellular scaffolds, where the frame method did not compromise mechanical strength, but it reduced inflammation significantly.

The effects of oral nutritional formula enriched with arginine, omega 3 fatty acids and nucleotides on methotrexate-induced experimental intestinal mucositis

The aim of this study to investigate the effect of an immunonutritional (IN) oral formula enriched with arginine, omega-3 fatty acids and nucleotides in methotrexate (MTX)-induced experimental intestinal mucositis model. In the study, 32 rats were divided into four groups consisting eight animals in each. Control group fed for 5 days with only saline with gavage, IN group fed for 5 days with an oral IN formula three times a day, MTX group with an intraperitoneal single dose MTX (20 mg/kg), followed by saline with gavage, MTX-IN group with a single dose of MTX, followed by an oral IN formula three times a day. The blood and jejunal tissue sample were collected and then the rats were sacrificed on the sixth day of the study. The level of TNF-α, IL-1β in serum, luminol, lusigenin, glutathione, myeloperoxidase, malondialdehyde and Na-K+ATPase in the jejunal tissue samples were analyzed. Histopathological examination was performed in the jejunal tissue samples. In the MTX group, TNF-α, IL-1β levels in serum, and luminol, lusigenin, malondialdehyde levels, and myeloperoxidase activity in tissue samples were found significantly higher than the control group. Glutathione and Na-K+ATPase levels were lower in the jejunal tissue of the MTX group compared to control group. However, the supplementation of IN with the MTX resulted in a significant increase in glutathione and Na-K+ATPase levels. Severe epithelium loss and inflammatory cell increase were observed in the MTX group on histological examination, whereas these parameters were regressed in the MTX-IN group. Increasing in the mitosis rate of enterocytes and inflammatory cell density decreased with the IN. In conclusion, this study shows that chemotherapy has adverse effects on intestinal mucosa and the IN formula has a protective effect of on MTX-associated intestinal damage.

Protective effects of agomelatine on testicular damage caused by bortezomib

Bortezomib is a chemotherapeutic agent used to treat several cancers; however, it exhibits severe side effects in testicular tissue. We investigated the use of agomelatine to prevent testicular tissue damage caused by bortezomib. We used 36 male Sprague-Dawley rats divided randomly into six equal groups: group 1, no treatment control; group 2, agomelatine treatment only; group 3, bortezomib treatment only for 48 h; group 4, bortezomib + agomelatine treatment for 48 h; group 5, bortezomib treatment only for 72 h; and group 6, bortezomib + agomelatine treatment for 72 h. After treatments, the rats were sacrificed and testicular tissue was harvested. Lipid oxidation (LPO) and superoxide dismutase (SOD) levels in the tissues were determined using biochemical methods. Tissue samples also were examined using histopathological and immunohistochemical techniques. The LPO level was increased, while the SOD level was decreased in the bortezomib treated groups. We found that agomelatine treatment normalized LPO and SOD activities in the bortezomib treated groups. In the spermatogonia and Sertoli cells, the staining density of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and caspase 3 were decreased in the bortezomib + agomelatine groups at both 48 and 72 h compared to bortezomib only treated groups. We observed maturation arrest, basal membrane thickening, increase in inflammatory cells and connective tissue, and edema between germ cells in the bortezomib only treated groups. By contrast, normal basal membrane, less edema and more normal maturation were observed in the bortezomib + agomelatine groups at 48 and 72 h. We found that agomelatine reduced the damaging effects of bortezomib. The use of agomelatine to prevent bortezomib induced testicular tissue damage in human patients should be investigated further.

Red Maca (Lepidium meyenii), a Plant from the Peruvian Highlands, Promotes Skin Wound Healing at Sea Level and at High Altitude in Adult Male Mice.

Nuñez, Denisse, Paola Olavegoya, Gustavo F. Gonzales, and Cynthia Gonzales-Castañeda. Red maca (Lepidium meyenii), a plant from the Peruvian highlands, promotes skin wound healing at sea level and at high altitude in adult male mice. High Alt Med Biol 18:373-383, 2017.-Wound healing consists of three simultaneous phases: inflammation, proliferation, and remodeling. Previous studies suggest that there is a delay in the healing process in high altitude, mainly due to alterations in the inflammatory phase. Maca (Lepidium meyenii) is a Peruvian plant with diverse biological properties, such as the ability to protect the skin from inflammatory lesions caused by ultraviolet radiation, as well as its antioxidant and immunomodulatory properties. The aim of this study was to determine the effect of high altitude on tissue repair and the effect of the topical administration of the spray-dried extract of red maca (RM) in tissue repair. Studies were conducted in male Balb/c mice at sea level and high altitude. Lesions were inflicted through a 10 mm-diameter excisional wound in the skin dorsal surface. Treatments consisted of either (1) spray-dried RM extract or (2) vehicle (VH). Animals wounded at high altitude had a delayed healing rate and an increased wound width compared with those at sea level. Moreover, wounding at high altitude was associated with an increase in inflammatory cells. Treatment with RM accelerated wound closure, decreased the level of epidermal hyperplasia, and decreased the number of inflammatory cells at the wound site. In conclusion, RM at high altitude generate a positive effect on wound healing, decreasing the number of neutrophils and increasing the number of macrophages in the wound healing at day 7 postwounding. This phenomenon is not observed at sea level.

Placental Malaria: A New Insight into the Pathophysiology.

Malaria in pregnancy poses a great health risk to mother and her fetus and results into complications, such as abortion, still birth, intra uterine growth retardation, and low birth weight. The heavy infiltration of Plasmodium falciparum-infected RBCs in the intervillous spaces of placenta seems to be responsible for all the complications observed. Infected RBCs in the placenta cause an inflammatory environment with increase in inflammatory cells and cytokines which is deleterious to the placenta. Increased inflammatory responses in the infected placenta result into oxidative stress that in turn causes oxidative stress-induced placental cell death. Moreover, heat shock proteins that are produced in high concentration in stressed cells to combat the stress have been reported in fewer concentrations in malaria-infected placenta. Pathologies associated with placental malaria seems to be the effect of a change in immune status from antibody-mediated immune response to cell-mediated immune response resulting into excess inflammation, oxidative stress, apoptosis, and decreased heat shock protein expression. However, we also need to study other aspects of pathologies so that better drugs can be designed with new molecular targets.

Comparison of early response to combined chemotherapy gemcitabine-cisplatin in non-small cell lung cancer animal model between using 18F-fluorodeoxyglucose and 18F-fluorothymidine

Non-small cell lung cancer (NSCLC) accounts for more than 80% of lung cancer. Nowadays, gemcitabine and cisplatin in combination have been adopted as the first-line chemotherapy for patients with NSCLC. This study aimed to monitor early response to combined chemotherapy of gemcitabine plus cisplatin in a mouse model of NSCLC by using 18F-fluorodeoxyglucose and 18F-fluorothymidine small animal positron emission tomography (PET). Lewis lung carcinoma-bearing C57BL/6 mice were treated with gemcitabine-cisplatin or saline. Small animal PET with 18F-FDG and 18F-FLT was performed before (baseline) and after treatment (on Day 3), respectively. Imaging results were confirmed by histopathological studies (hematoxylin and eosin staining, Ki67 staining). Compared to the results in the control group, gemcitabine-cisplatin in the treated group significantly inhibited tumor growth ( P<0.05). In the treated group, the maximum standardized uptake value (SUVmax) of 18F-FLT decreased significantly from 0.59±0.05 (baseline) to 0.28±0.05 (Day 3) ( P<0.05). There was no significant difference between baseline (4.35±0.46) and that on Day 3 (4.02±0.47) on 18F-FDG SUVmax ( P>0.05). The proliferation of tumor assessed by Ki67 staining decreased significantly after treatment of one dose of gemicitabine-cisplatin ( P<0.05). The staining of HE showed an increase in necrotic and inflam- matory cells after the treatment. This study demonstrated that the uptake of 18F-FLT reduced more rapidly and signi-ficantly than that of 18F-FDG and was less disturbed by the increase of inflammatory cells after chemotherapy.

Proteasomes in corneal epithelial cells and cultured autologous oral mucosal epithelial cell sheet (CAOMECS) graft used for the ocular surface regeneration

This study focuses on characterizing proteasomes in corneal epithelial cells (CEC) and in cultured autologous oral mucosal epithelial cell sheets (CAOMECS) used to regenerate the ocular surface. Limbal stem cell deficiency (LSCD) was surgically induced in rabbit corneas. CAOMECS was engineered and grafted onto corneas with LSCD to regenerate the ocular surface. LSCD caused an increase in inflammatory cells in the ocular surface, an increase in the formation of immunoproteasomes (IPR), and a decrease in the formation of constitutive proteasome (CPR). Specifically, LSCD-diseased CEC (D-CEC) showed a decrease in the CPR chymotrypsin-like, trypsin-like and caspase-like activities, while healthy CEC (H-CEC) and CAOMECS showed higher activities. Quantitative analysis of IPR inducible subunit (B5i, B2i, and B1i) were performed and compared to CPR subunit (B5, B2, and B1) levels. Results showed that ratios B5i/B5, B2i/B2 and B1i/B1 were higher in D-CEC, indicating that D-CEC had approximately a two-fold increase in the amount of IPR compared to CAOMECS and H-CEC. Histological analysis demonstrated that CAOMECS-grafted corneas had a re-epithelialized surface, positive staining for CPR subunits, and weak staining for IPR subunits. In addition, digital quantitative measurement of fluorescent intensity showed that the CPR B5 subunit was significantly more expressed in CAOMECS-grafted corneas compared to non-grafted corneas with LSCD. CAOMECS grafting successfully replaced the D-CEC with oral mucosal epithelial cells with higher levels of CPR. The increase in constitutive proteasome expression is possibly responsible for the recovery and improvement in CAOMECS-grafted corneas.