scholarly journals Quantitative PCR Analysis of the Bartonella henselae carD Gene during Bacterial Stress

2021 ◽  
Author(s):  
◽  
Jacob Simmonds
Keyword(s):  
Life Cycle ◽  
Stress Response ◽  
Quantitative Pcr ◽  
Bartonella Henselae ◽  
Bacterial Species ◽  
Clinical Manifestations ◽  
Stress Conditions ◽  
Bartonella Species ◽  
Organ Systems ◽  
Significant Difference

<p>The Bartonella genus is comprised of arthropod-borne, intracellular bacterial pathogens that colonise the mammalian bloodstream. A large number of mammalian species are hosts for one or more Bartonella species, as either reservoir or incidental hosts. Bartonella species are only able to invade and replicate in host red blood cells in the reservoir host, and to be taken up by an associated haematophagous arthropod vector to complete transmission and the bacterial life cycle. Humans are the reservoir hosts for B. quintana and B. bacilliformis, and are incidental hosts for more than 16 additional zoonotic Bartonella species, including B. henselae, which is normally carried by cats. B. henselae infection, usually acquired through cat scratches or bites, can result in several clinical manifestations, with varying degrees of severity; the most common of these is cat scratch disease, where symptoms commonly range from enlarged lymph nodes to severe fever. Although usually a mild illness, B. henselae infection can occasionally lead to severe symptoms, affecting neurological and other major organ systems.  During their life cycle the Bartonellae must adapt to various toxic host environments; such adaptation is mediated by several bacterial stress pathways, which modify bacterial transcription. However, many gaps remain in the understanding of B. henselae stress response pathways. The object of this study, the carD gene, was identified as a possible component of the Bartonella stress response. The carD gene has been shown to be critical for stress defence in other bacterial species, including Mycobacterium tuberculosis, Thermus thermophilus, and Myxococcus xanthus.  Our study aimed to investigate whether carD played as significant a role during the B. henselae response to stresses as it does in other bacterial genera. We first attempted to perform growth comparisons between a B. henselae carD mutant strain and a wild type strain during exposure to stress conditions; however, our mutagenic carD plasmid interfered with bacterial growth of Escherichia coli cultures, which hindered transformation and generation of a B. henselae carD mutant. As an alternative, we investigated the expression of B. henselae carD under stress conditions, comparing carD expression during stress against a non-stressed B. henselae control, using quantitative PCR. We found no significant difference of expression of the carD gene between the control and any of our conditions, although a trend of increased carD expression was found in several stress conditions. We believe that these findings merit further study into the role of carD in the B. henselae stress response.</p>

scholarly journals Quantitative PCR Analysis of the Bartonella henselae carD Gene during Bacterial Stress

2021 ◽  
Author(s):  
◽  
Jacob Simmonds
Keyword(s):  
Life Cycle ◽  
Stress Response ◽  
Quantitative Pcr ◽  
Bartonella Henselae ◽  
Bacterial Species ◽  
Clinical Manifestations ◽  
Stress Conditions ◽  
Bartonella Species ◽  
Organ Systems ◽  
Significant Difference

<p>The Bartonella genus is comprised of arthropod-borne, intracellular bacterial pathogens that colonise the mammalian bloodstream. A large number of mammalian species are hosts for one or more Bartonella species, as either reservoir or incidental hosts. Bartonella species are only able to invade and replicate in host red blood cells in the reservoir host, and to be taken up by an associated haematophagous arthropod vector to complete transmission and the bacterial life cycle. Humans are the reservoir hosts for B. quintana and B. bacilliformis, and are incidental hosts for more than 16 additional zoonotic Bartonella species, including B. henselae, which is normally carried by cats. B. henselae infection, usually acquired through cat scratches or bites, can result in several clinical manifestations, with varying degrees of severity; the most common of these is cat scratch disease, where symptoms commonly range from enlarged lymph nodes to severe fever. Although usually a mild illness, B. henselae infection can occasionally lead to severe symptoms, affecting neurological and other major organ systems.  During their life cycle the Bartonellae must adapt to various toxic host environments; such adaptation is mediated by several bacterial stress pathways, which modify bacterial transcription. However, many gaps remain in the understanding of B. henselae stress response pathways. The object of this study, the carD gene, was identified as a possible component of the Bartonella stress response. The carD gene has been shown to be critical for stress defence in other bacterial species, including Mycobacterium tuberculosis, Thermus thermophilus, and Myxococcus xanthus.  Our study aimed to investigate whether carD played as significant a role during the B. henselae response to stresses as it does in other bacterial genera. We first attempted to perform growth comparisons between a B. henselae carD mutant strain and a wild type strain during exposure to stress conditions; however, our mutagenic carD plasmid interfered with bacterial growth of Escherichia coli cultures, which hindered transformation and generation of a B. henselae carD mutant. As an alternative, we investigated the expression of B. henselae carD under stress conditions, comparing carD expression during stress against a non-stressed B. henselae control, using quantitative PCR. We found no significant difference of expression of the carD gene between the control and any of our conditions, although a trend of increased carD expression was found in several stress conditions. We believe that these findings merit further study into the role of carD in the B. henselae stress response.</p>

Assessment of Bacterial Contamination of Orthodontic Arch wire

2019 ◽  
Vol 31 (1) ◽  
pp. 48-51
Author(s):  
Suha S Hassan ◽  
Nidhal H. Ghaib ◽  
Batool H Al-Ghurabi
Keyword(s):  
Bacterial Contamination ◽  
Microbial Contamination ◽  
Ethylenediaminetetraacetic Acid ◽  
Bacterial Species ◽  
Brain Heart Infusion ◽  
Control Groups ◽  
Dental Practitioners ◽  
Significant Difference ◽  
Bacillus Spp ◽  
Droplet Transmission

Background: The microorganisms can impend the life of health care professional and particularly the dental practitioners. They can be transmitted by different ways like airborne and droplet transmission. The current study was carried out to identify whether the arch wires that received from the manufactures are free from microbial contamination and to determine the bacterial species attached to the arch wires. Materials and Methods: This study involved eighty samples, consisted of two types of arch wires (nitinol and stainless-steel) from four companies (3M, G&H, Jiscop, OrthoTechnology). These wires inserted in a plane tube that contains 10 -ml of (Tris [tris(hydroxymethyl)aminomethane] and EDTA (ethylenediaminetetraacetic acid) tris-EDTA and brain heart infusion (BHI) broth. A 0.1 ml was withdrawn from the tube and spread on agar plates. The control groups consist of 16 plane tube (8 tubes with tris-EDTA and other 8 tubes with (BHI). Results: Microbial sampling yielded growth from 5 of the 80 arch wires. The predominant bacteria that isolated were Bacillus spp. No growth was recovered from 75 of the samples and from controls. The bacteria were isolated by BHI reagent and no growth was observed by tris-EDTA reagent with statistically significant difference (P<0.05). The Bacillus spp. found only in the G&H and Jiscop companies, however, no statistically significant difference was found among them (P>0.05). With regard to the presence and distribution of bacteria according to the types of wires, the present results clarified that cases of contamination with Bacillus spp. were found in the nitinol arch wires with statistically significant difference (P<0.05). Conclusions: The results of the current study revealed low count of bacterial contamination in the two types of companies (G&H and Jiscop). Not all materials that received from the manufactures are free from contamination and an effective sterilization regimen is needed to avoid cross-contamination.

PRECLINICAL STUDY OF THE VETERINARY DRUGS TOXICITY

1970 ◽  
pp. 61-64
Author(s):  
E. K. Rakhmatullin ◽  
O. D. Sklyarov
Keyword(s):  
Lethal Dose ◽  
Clinical Manifestations ◽  
Preclinical Study ◽  
Veterinary Drugs ◽  
Toxic Effects ◽  
Laboratory Animals ◽  
Therapeutic Effects ◽  
Significant Information ◽  
Important Indicator ◽  
Organ Systems

Preclinical study of the drugs toxicity was analysed it allows predicting the safety of veterinary drugs in laboratory animals. The fundamental normative instruments in the field of preclinical study of drugs for veterinary medicine and animal husbandry are Order of the Ministry of Agriculture of the Russian Federation dated 06.03.2018 N 101 and GOST 33044-2014 Principles of Good Laboratory Practice. An important indicator of the preclinical study of the veterinary drugs is the determination (calculation) of median lethal dose value (lethal dose for half of the animals tested) or concentration (LD50 or LC50). Existing methods for determining this indicator make it possible at the initial study stage to determine the degree and class the drug of toxicity. Studying the symptoms of intoxication in the analysis of pharmacological substances one obtains significant information about the nature of the action of the future drug. The clinical manifestations of intoxication with damage to various organ systems are presented. As criteria for assessing the toxic effects of veterinary drugs it is recommended to determine LD50, cumulation coefficient, latitude index of therapeutic effects, dose level of toxic effects in the experiment which allows predicting the nature and degree of toxic effects of the drug even at the stage of preclinical veterinary drugs study.

THE CLINICAL, LABORATORY MANIFESTATIONS AND HISTOPATHOLOGY IN NEPHROTIC PATIENTS WITH LUPUS NEPHRITIS

2018 ◽  
pp. 52-58
Author(s):  
Le Thuan Nguyen ◽  
Bui Bao Hoang
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Lupus Nephritis ◽  
Lupus Erythematosus ◽  
Clinical Laboratory ◽  
Activity Index ◽  
Clinical Manifestations ◽  
Systemic Lupus ◽  
Cross Sectional ◽  
Organ Systems ◽  
Tubulointerstitial Lesions

Introduction: Systemic lupus erythematosus (SLE) is an autoimmune disease involving multiple organ systems. The kidney appears to be the most commonly affected organ, especially nephrotic is a serious kidney injury. The clinical, laboratory manifestations and histopathology are very useful for diagnosis, provide the means of predicting prognosis and guiding therapy in nephrotic patients with lupus nephritis. Methods: Descriptive cross-sectional study of nephrotic patients with lupus treated in the Department of Nephrology Trung Vuong Hospital and Cho Ray Hospital between May/2014 and May/2017. Renal histopathological lesions were classified according to International Society of Nephrology/Renal Pathology Society - ISN/RPS ’s 2003. The clinical, laboratory manifestations and histopathological features were described. Results: Of 32 LN with nephritic range proteinuria cases studied, 93.7% were women. The 3 most common clinical manifestations were edema (93.8%), hypertension (96.8%) and pallor (68.9%), musculoskeletal manifestions (46.9%), malar rash (40.6%). There was significant rise in laboratory and immunological manifestions with hematuria (78.1%), Hb < 12g/dL (93.5%), increased Cholesterol (100%), and Triglycerid (87.5%), Creatinine > 1.4 mg/dL (87.5%), increased BUN 71.9%, ANA (+) 93.8%, Anti Ds DNA(+) 96.9%, low C3: 96.9%, low C4: 84.4%. The most various and severe features were noted in class IV with active tubulointerstitial lesions and high activity index. Conclusion: Lupus nephritis with nephrotic range proteinuria has the more severity of histopathological feature and the more severity of the more systemic organ involvements and laboratory disorders were noted. Key words: Systemic lupus, erythematosus (SLE) lupus nepphritis, clinical

scholarly journals Platelet Depletion is Effective in Ameliorating Anxiety-Like Behavior and Reducing the Pro-Inflammatory Environment in the Hippocampus in Murine Experimental Autoimmune Encephalomyelitis

2019 ◽  
Vol 8 (2) ◽  
pp. 162 ◽  
Author(s):  
Pece Kocovski ◽  
Xiangrui Jiang ◽  
Claretta D’Souza ◽  
Zhenjiang Li ◽  
Phuc Dang ◽  
...  
Keyword(s):  
Experimental Autoimmune Encephalomyelitis ◽  
Neuropsychiatric Symptoms ◽  
Critical Role ◽  
Clinical Manifestations ◽  
Lymphocytic Infiltration ◽  
Disease Stage ◽  
Autoimmune Encephalomyelitis ◽  
Significant Difference ◽  
Platelet Depletion ◽  
Experimental Autoimmune

The neuropsychiatric symptoms of multiple sclerosis (MS), such as anxiety and depression, can result from disease activity itself as well as psychological reaction to an unfavorable diagnosis. Accordingly, the literature reports evidence of increased anxiety-like behavior in experimental autoimmune encephalomyelitis (EAE), an accepted MS model. Due to the recently described critical role of platelets in inflammation and autoimmune disease, we examined the relationship between platelets, inflammation, and anxiety-like behavior in EAE. In the elevated plus maze, EAE-induced C57BL/6J mice showed decreased time spent in the open arms relative to vehicle-only controls, demonstrating an increase in anxiety-like behavior. This effect occurred in the presence of platelet–neuron association, but absence of lymphocytic infiltration, in the hippocampal parenchyma. Platelet depletion at the pre-clinical disease stage, using antibody-mediated lysis prevented the EAE-induced increase in anxiety-like behavior, while no significant difference in distance moved was recorded. Furthermore, platelet depletion was also associated with reduction of the pro-inflammatory environment to control levels in the hippocampus and prevention of EAE disease symptomology. These studies demonstrate the high efficacy of a platelet-targeting approach in preventing anxiety-like symptoms and clinical manifestations of EAE and have implications for the treatment of neuropsychiatric symptoms in MS.

scholarly journals Reassessing the Role of the Active TGF-β1 as a Biomarker in Systemic Sclerosis: Association of Serum Levels with Clinical Manifestations

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Andréa Tavares Dantas ◽  
Sayonara Maria Calado Gonçalves ◽  
Anderson Rodrigues de Almeida ◽  
Rafaela Silva Guimarães Gonçalves ◽  
Maria Clara Pinheiro Duarte Sampaio ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Clinical Manifestations ◽  
Serum Levels ◽  
Vascular Involvement ◽  
Digital Ulcers ◽  
Serum Samples ◽  
Peripheral Blood Mononuclear ◽  
Pbmc Culture ◽  
Significant Difference ◽  
Skin Biopsies

Objective. To determine active TGF-β1 (aTGF-β1) levels in serum, skin, and peripheral blood mononuclear cell (PBMC) culture supernatants and to understand their associations with clinical parameters in systemic sclerosis (SSc) patients.Methods. We evaluated serum samples from 56 SSc patients and 24 healthy controls (HC). In 20 SSc patients, we quantified spontaneous or anti-CD3/CD28 stimulated production of aTGF-β1 by PBMC. The aTGF-β1 levels were measured by ELISA. Skin biopsies were obtained from 13 SSc patients and six HC, and TGFB1 expression was analyzed by RT-PCR.Results. TGF-β1 serum levels were significantly higher in SSc patients than in HC (p< 0.0001). Patients with increased TGF-β1 serum levels were more likely to have diffuse subset (p= 0.02), digital ulcers (p= 0.02), lung fibrosis (p< 0.0001), positive antitopoisomerase I (p= 0.03), and higher modified Rodnan score (p= 0.046). Most of our culture supernatant samples had undetectable levels of TGF-β1. No significant difference in TGFB1 expression was observed in the SSc skin compared with HC skin.Conclusion. Raised active TGF-β1 serum levels and their association with clinical manifestations in scleroderma patients suggest that this cytokine could be a marker of fibrotic and vascular involvement in SSc.

scholarly journals 2-Piece Cork Stoppers as Alternative for Valorization of Thin Cork Planks: Analysis by LCA Methodology

Foods ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 873
Author(s):  
Francisco Javier Flor-Montalvo ◽  
Agustín Sánchez-Toledo Ledesma ◽  
Eduardo Martínez Cámara ◽  
Emilio Jiménez-Macías ◽  
Jorge Luis García-Alcaraz ◽  
...  
Keyword(s):  
Life Cycle Assessment ◽  
Life Cycle ◽  
Raw Materials ◽  
Lca Methodology ◽  
Significant Difference ◽  
Different Dimensions ◽  
The Impact ◽  
Different Thickness ◽  
Cork Stoppers ◽  
Wine Bottle

Natural stoppers are a magnificent closure for the production of aging wines and unique wines, whose application is limited by the availability of raw materials and more specifically of cork sheets of different thickness and quality. The growing demand for quality wine bottle closures leads to the search for alternative stopper production. The two-piece stopper is an alternative since it uses non-usable plates in a conventional way for the production of quality caps. The present study has analyzed the impact of the manufacture of these two-piece stoppers using different methodologies and for different dimensions by developing an LCA (Life Cycle Assessment), concluding that the process phases of the plate, its boiling, and its stabilization, are the phases with the greatest impact. Likewise, it is detected that the impacts in all phases are relatively similar (for one kg of net cork produced), although the volumetric difference between these stoppers represents a significant difference in impacts for each unit produced.

scholarly journals AB0345 Th9 CELLS AND THEIR ASSOCIATED CYTOKINES: THE PROBABLE PLAYERS IN SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) AND TYPE 1 DIABETES MELLITUS (T1DM)

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 1197.2-1198
Author(s):  
N. Mohannad ◽  
M. Moaaz ◽  
R. Mohamed Shehata
Keyword(s):  
T Cells ◽  
T Cell ◽  
Disease Activity ◽  
Clinical Manifestations ◽  
Moderate Activity ◽  
Healthy Controls ◽  
Group I ◽  
Pancreatic Islets Of Langerhans ◽  
Th9 Cells ◽  
Significant Difference

Background:SLE is an autoimmune disease (AID) of unknown origin. Several factors can contribute to immune dysfunction in SLE.Interleukin 9 (IL9) is a newly emerging T cell-derived factor preferentially expressed by CD4+T cells: T helper 9 (Th9)IL9 targets different cell lineages, and can contribute to the development of allergic & AIDsWhether abnormal expression and secretion of IL9 are present in SLE patients (pts) still unidentified. It is also unclear whether IL9 exerts main proinflammatory or anti-inflammatory activities in SLE. T1DM is characterized by inflammation of the pancreatic islets of Langerhans. Insulitis progresses over time and β cells become destroyed then clinical DM is established. T1DM is regarded as a T cell-driven AIDObjectives:Evaluation of the expression of CD4+ IL9+ T cells & the level of IL9 in SLE pts compared to both healthy subjects & pts with another AID: T1DM.Also, to evaluate the correlation of these expressions with clinical features, laboratory parameters and SLE activityMethods:The study included: Group I 25 SLE pts fulfilling SLICC classification criteria divided into 2 subgroups (gps) according to SLE disease activity index (SLEDAI) IA: 20 pts with mild to moderate activity (<12) IB:5 pts with severe activity (>12) recruited from rheumatology clinic or internal medicine ward (Rheumatology unit), Main University Hospital, Alexandria. Group II 15 healthy individuals as a first control gp. Group III 15 pts with T1DM fulfilling the American Diabetes Association criteria as a second control gp. All pts were subjected to history taking, clinical examination,laboratory investigations: CBC,LFT,KFT,ESR,CRP,ANA,Anti-dsDNA,Th9 cell expression detection by flowcytometry and serum IL9 by ELISAResults:There was no statistical difference between all gps as regards age & sex but a significant increased ESR in SLE compared to controls & T1DM p< 0.001 p=0.001Th9 expression was highly increased in SLE pts, range 0.13-4.54% & mean ±SD=1.50 ± 1.47% than both control gps. In healthy controls Th9 ranged between 0.0-1.29% with mean 0.37 ±0.52%, while in T1DM pts ranged between 0.03 to 2.13% with mean of 0.67 ± 0.59%. A high significant difference was found between SLE pts and controls p=0.001, an insignificant rise was seen in SLE pts compared to T1DM pts p=0.157. A high significant increase in Th9 was found in severe SLE: mean of 3.74 ±1.15% than in pts with mild to moderate SLE: mean 0.94±0.88% p=<0.001IL9 level was highly increased in SLE pts: mean of 42.83± 23.98 pg/ml than both control gps. In healthy controls the mean was 8.54±13.27, while in T1DM with mean of 29.17±16.09 pg/ml. A high significant difference was found between SLE pts and normal controls p<0.001 but an insignificant rise with T1DM p=0.294. A high significant increase in IL9 in pts with severe ds compared to mild to moderate pts p<0.001.A significant direct correlation between Th9 & IL9 and SLEDAI/105 A significant direct correlation between damage index and Th9 p=0.040 but not IL9 p=0.053In SLE no significant relation between Th9 or IL9 & clinical manifestations or disease duration. A direct correlations between Th9 & ESR p=0.046 and CRP p=0.025,a significant correlation between IL9 and CRP p=0.033, no correlations between Th9&IL9 level and anti-dsDNA p=0.593& 0.4 Significant direct correlation between Th9 and IL9 in T1DM pts, still no correlation with glycemic profile. IL9 levels were significantly increased in SLE with elevated CRP p=0.033 & the % of Th9 cells were increased with elevated ESR and CRP p=0.025, 0.046Conclusion:In SLE pts; IL9 level and Th9 cells expression were significantly elevated compared to healthy controls. IL9 levels and the percentages of Th9 directly correlated with the SLE disease activity. IL9 levels also were significantly increased in T1DM pts compared to controls,but they were less expressed than in SLE. This suggests an important role of IL9 in the pathogenesis AIDs as SLEReferences:[1]Tahernia L et al. Cytokines in SLE: their role in pathogenesis of disease and possible therapeutic opportunities. Rheum Res 2017Disclosure of Interests:None declared

scholarly journals Flavonoids: Potential Candidates for the Treatment of Neurodegenerative Disorders

Biomedicines ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 99
Author(s):  
Shweta Devi ◽  
Vijay Kumar ◽  
Sandeep Kumar Singh ◽  
Ashish Kant Dubey ◽  
Jong-Joo Kim
Keyword(s):  
Stress Response ◽  
Stress Responses ◽  
Neurodegenerative Disorders ◽  
Cell Damage ◽  
Cellular Stress ◽  
Clinical Manifestations ◽  
Cellular Stress Response ◽  
Dramatic Rise ◽  
Cellular Stress Responses

Neurodegenerative disorders, such as Parkinson’s disease (PD), Alzheimer’s disease (AD), Amyotrophic lateral sclerosis (ALS), and Huntington’s disease (HD), are the most concerning disorders due to the lack of effective therapy and dramatic rise in affected cases. Although these disorders have diverse clinical manifestations, they all share a common cellular stress response. These cellular stress responses including neuroinflammation, oxidative stress, proteotoxicity, and endoplasmic reticulum (ER)-stress, which combats with stress conditions. Environmental stress/toxicity weakened the cellular stress response which results in cell damage. Small molecules, such as flavonoids, could reduce cellular stress and have gained much attention in recent years. Evidence has shown the potential use of flavonoids in several ways, such as antioxidants, anti-inflammatory, and anti-apoptotic, yet their mechanism is still elusive. This review provides an insight into the potential role of flavonoids against cellular stress response that prevent the pathogenesis of neurodegenerative disorders.

scholarly journals POS0369 ELEVATED EXPRESSION OF TIM-3 ON NEUTROPHILS CORRELATES WITH DISEASE ACTIVITY AND SEVERITY OF ANKYLOSING SPONDYLITIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 414.2-415
Author(s):  
X. Huang ◽  
T. W. Li ◽  
J. Chen ◽  
Z. Huang ◽  
S. Chen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Ankylosing Spondylitis ◽  
Disease Activity ◽  
Immune Cells ◽  
Clinical Manifestations ◽  
Innate Immune ◽  
Innate Immune Cells ◽  
Bacterial Killing ◽  
Markers Of Inflammation ◽  
Significant Difference

Background:Ankylosing spondylitis (AS) is a type of common, chronic inflammatory disease that compromises the axial skeleton and sacroiliac joints, causing inflammatory low back pain and progressive spinal stiffness, over time some patients develop spinal immobility and ankylosis which can lead to a decrease in quality of life. The last few decades, evidence has clearly indicated that neutrophil also plays key roles in the progression of AS. However, the immunomodulatory roles and mechanisms of neutrophils in AS are poorly understood. T-cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) has been reported as an important regulatory molecule, expressed and regulated on different innate immune cells, plays a pivotal role in several autoimmunity diseases. Recent study indicates that Tim3 is also expressed on neutrophils. However, the frequency and roles of Tim3-expressing neutrophils in AS was not clear.Objectives:In this study, we investigated the expression of Tim3 on neutrophils in AS patients and explored the correlation between the level of Tim3-expressing neutrophils and the disease activity and severity of AS.Methods:Patients with AS were recruited from Guangdong Second Provincial General Hospital (n=62). Age/sex-matched volunteers as Healthy controls (HC) (n=39). The medical history, clinical manifestations, physical examination, laboratory measurements were recorded. The expression of costimulatory molecules including programmed death 1 (PD-1), Tim-3 on neutrophils were determined by flow cytometry. The mRNA expression of PD-1 and Tim-3 was determined by real-time PCR. The levels of Tim3-expressing neutrophils in AS patients were further analyzed for their correlation with the markers of inflammation such as ESR,CRP,WBC and neutrophil count(NE), as well as disease activity and severity of AS. The expression of Tim3 on neutrophils was monitored during the course of treatment (4 weeks).Results:The expression of Tim3 on neutrophils in patients with AS was increased compared to the HC (Figure 1A). However, significant difference was observed in the frequency of PD-1-expressing neutrophils between AS patients and HC (Figure 1B). The expression analysis of Tim-3 mRNA, but not PD-1, confirmed the results obtained from flow cytometry (Figure 1C). The level of Tim3-expressing neutrophils in patients with AS showed an positive correlation with ESR, CRP and ASAS-endorsed disease activity score (ASDAS) (Figure 1D). Moreover, the frequency of Tim3-expressing neutrophils in active patients(ASDAS≥1.3) was increased as compare with the inactive patients (ASDAS<1.3) (Figure 1E). As shown in Figure 1F, the frequency of Tim3-expressing neutrophils decreased after the treatment.Conclusion:Increased Tim-3 expression on neutrophils may be a novel indicator to assess disease activity and severity in AS, which may serves as a negative feedback mechanism preventing potential tissue damage caused by excessive inflammatory responses in AS patients.References:[1]Han, G., Chen, G., Shen, B. & Li, Y., Tim-3: an activation marker and activation limiter of innate immune cells. FRONT IMMUNOL 4 449 (2013).[2]Vega-Carrascal, I. et al., Galectin-9 signaling through TIM-3 is involved in neutrophil-mediated Gram-negative bacterial killing: an effect abrogated within the cystic fibrosis lung. J IMMUNOL 192 2418 (2014).Figure 1.(A,B)The expression of Tim3 and PD-1 on neutrophils in AS and HC were determined by flow cytometry.(C) The expression of Tim3 and PD-1 on neutrophils in AS and HC were determined by RT-PCR.(D)The correction between Tim3-expressing neutrophils and ESR,CRP,ASDAS.(E) The expression of Tim3 on neutrophils in active and inactive patients.(F) Influence of treatment on the frequency of Tim3-expressing neutrophils.Disclosure of Interests:None declared

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