Cytoplasm Of Neutrophils Research Articles

Exceptional microorganisms on blood smear.

A 67-year-old woman with metastatic pancreatic adenocarcinoma was referred for acute respiratory distress syndrome and severe sepsis. She had a totally implantable central venous access device for chemotherapy administration and home parenteral nutrition. Laboratory workup at admission revealed increased C-reactive protein (209 mg/L), mild normocytic anemia (hemoglobin 8.6 g/dL), and severe acute thrombocytopenia (platelet count 3 × 109/L, as compared to 257 × 109/L 8 days before). White blood cell count, absolute neutrophil count, and blood smear were normal. Laboratory tests excluded disseminated intravascular coagulation. The patient was treated with cefotaxime and metronidazole. Forty-eight hours later, blood smear examination done on a sample taken through the intravenous line revealed yeasts (Image 1, panels A-E-B-D). Moreover, some bacilli were also observed (Image 1, panels C-E). Yeasts and budding yeasts in monocyte and neutrophil cytoplasm, extracellular branching pseudohyphae (arrow, respectively panels A-E-B-D May–Grünwald–Giemsa stain, original magnification 100× and 20×). Extra- and intracellular bacilli (arrowhead, respectively panels C-E, May–Grünwald–Giemsa stain, original magnification 100×) This exceptional observation of microorganisms on blood smear led to microbiological tests, identifying Candida albicans and Klebsiella pneumoniae. Severe thrombocytopenia was most likely related to severe sepsis. The patient was admitted to a palliative care unit. Nothing to report.

Paquimeningitis y glomerulonefritis crescéntica en vasculitis p-ANCA: Reporte de caso

Las vasculitis asociadas a anticuerpos contra el citoplasma del neutrófilo configuran patologías multisistémicas caracterizadas por afectar pequeños vasos. Se ha reportado compromiso de sistema nervioso central en 2% a 8% de los casos, rara vez al inicio del cuadro clínico.
 Se reporta el caso de un hombre de 67 años de edad con vasculitis p-ANCA, quien debutó con proptosis, paquimeningitis y disfagia severa. Posteriormente deterioro renal, necesidad de hemodiálisis, ciclofosfamida y recambio plasmático terapéutico con lo cual alcanza criterios de remisión parcial y egresa de diálisis. Las vasculitis ANCA se relacionan con una amplia gama de signos y síntomas inespecíficos. La edad de aparición, creatinina inicial y requerimiento de diálisis, son factores pronósticos. El inicio oportuno del tratamiento es fundamental, requiriendo por tanto un alto índice de sospecha

For fulminant rapidly progressive glomerulonephritis in a 17-year-old female patient with ANCA-associated vasculitis.

A case of rapidly progressive glomerulonephritis in a 17-year-old patient associated with antibodies against the cytoplasm of neutrophils (ANCA) vasculitis - ANCA-associated vasculitis is associated with antibodies to proteinase-3 and morphological picture extracapillar glomerulonephritis with sclerotic lesion of up to 80% of the glomeruli. The peculiarity of the case is the presence of morphologically confirmed when alloimmune rapidly progressive glomerulonephritis type III a pronounced glow-focal granular nature of immunoglobulin classes G and M on the basement membrane of capillaries. The appointment of immunosuppressive therapy led to a decrease in systemic manifestations of vasculitis, but there was a rapid increase in terminal renal failure, which required substitution therapy with hemodialysis. The possible mechanisms of the rapid-training course of the disease in the observed patient, prospects for kidney transplantation are discussed.

Linked help from bacterial proteins drives autoantibody production in small vessel vasculitis

The small vessel vasculitides granulomatosis with polyangiitis (GPA) and microscopic polyangiitis are associated with autoantibodies to neutrophil cytoplasm antigens (ANCA), principally proteinase-3 (PR3) and myeloperoxidase (MPO). There is an association between GPA and nasal carriage of Staphylococcus aureus. The recent finding that S. aureus produces proteins that bind tightly to and block the function of both PR3 and MPO suggests a mechanism for ANCA formation. The bacterial protein-autoantigen conjugate is recognised by B cells with ANCA specificity, internalised, and the bacterial protein processed and presented to T cells with specificity for bacterial peptides. The T cell can then provide help to the B cell, allowing class switching, affinity maturation and the production of pathogenic ANCA. This mechanism predicts that T cells with this specificity will be found in patients, and that the bacterial protein-autoantigen conjugate will be particularly efficient at eliciting ANCA production.

A framework for nucleus and overlapping cytoplasm segmentation in cervical cytology extended depth of field and volume images

We propose a framework to detect and segment nuclei and segment overlapping cytoplasm in cervical cytology images. This is a challenging task due to folded cervical cells with spurious edges, poor contrast of cytoplasm and presence of neutrophils and artifacts. The algorithm segments nuclei and cell clumps in extended depth of field (EDF) images and uses volume images to segment overlapping cytoplasm. The boundaries are first approximated by a defined similarity metric and are refined in two steps by reducing concavity, iterative smoothing and outliers removal. We evaluated our framework on two public datasets provided in the first and second overlapping cervical cell segmentation challenges (ISBI 2014 and 2015). The results show that our method outperforms other state-of-the-art algorithms on both datasets. The results on the ISBI 2014 dataset show that our method missed less than 5% of cells when the pairwise cell overlapping degree was not higher than 0.3 and it missed only 7% of cells on average in a dataset of 810 synthetic images with 4860 (overlapping) cells. On the same dataset, it outperforms other state-of-the-art methods in nucleus detection with precision 0.961 and recall 0.933. The results on the ISBI 2015 dataset containing real cervical EDF images show that our method misses around 20% of cells in EDF images where a segmentation is considered a miss if it has dice similarity coefficient not greater than 0.7. The 20% miss rate is around half of the miss rate of two other recent methods.

Synovial calprotectin

Recently, several synovial biomarkers have been introduced into the algorithm for the diagnosis of a prosthetic joint infection (PJI). Alpha defensin is a promising biomarker, with a high sensitivity and specificity, but it is expensive. Calprotectin is a protein that is present in the cytoplasm of neutrophils, is released upon neutrophil activation and exhibits anti-microbial activity. Our aim, in this study, was to determine the diagnostic potential of synovial calprotectin in the diagnosis of a PJI. In this pilot study, we prospectively collected synovial fluid from the hip, knee, shoulder and elbow of 19 patients with a proven PJI and from a control group of 42 patients who underwent revision surgery without a PJI. PJI was diagnosed according to the current diagnostic criteria of the Musculoskeletal Infection Society. Synovial fluid was centrifuged and the supernatant was used to measure the level of calprotectin after applying a lateral flow immunoassay. The median synovial calprotectin level was 991 mg/L (interquartile range (IQR) 154 to 1787) in those with a PJI and 11 mg/L (IQR 3 to 29) in the control group (p < 0.0001). Using a cut-off value of 50 mg/L, this level showed an excellent diagnostic accuracy, with an area under the curve of 0.94. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) was 89%, 90%, 81% and 95% respectively. The NPV was 97% in the nine patients with a chronic PJI. Synovial calprotectin may be a valuable biomarker in the diagnosis of a PJI, especially in the exclusion of an infection. With a lateral flow immunoassay, a relatively rapid quantitative diagnosis can be made. The measurement is cheap and is easy to use. Cite this article: Bone Joint J 2017;99-B:660-5.

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator

Neutrophils are crucial to host innate defense and, consequently, constitute an important area of medical research. The phagosome, the intracellular compartment where the killing and digestion of engulfed particles take place, is the main arena for neutrophil pathogen killing that requires tight regulation. Phagosomal pH is one aspect that is carefully controlled, in turn regulating antimicrobial protease activity. Many fluorescent pH-sensitive dyes have been used to visualize the phagosomal environment. S-1 has several advantages over other pH-sensitive dyes, including its dual emission spectra, its resistance to photo-bleaching, and its high pKa. Using this method, we have demonstrated that the neutrophil phagosome is unusually alkaline in comparison to other phagocytes. By using different biochemical conjugations of the dye, the phagosome can be delineated from the cytoplasm so that changes in the size and shape of the phagosome can be assessed. This allows for further monitoring of ionic movement.

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator.

Neutrophils are crucial to host innate defense and, consequently, constitute an important area of medical research. The phagosome, the intracellular compartment where the killing and digestion of engulfed particles take place, is the main arena for neutrophil pathogen killing that requires tight regulation. Phagosomal pH is one aspect that is carefully controlled, in turn regulating antimicrobial protease activity. Many fluorescent pH-sensitive dyes have been used to visualize the phagosomal environment. S-1 has several advantages over other pH-sensitive dyes, including its dual emission spectra, its resistance to photo-bleaching, and its high pKa. Using this method, we have demonstrated that the neutrophil phagosome is unusually alkaline in comparison to other phagocytes. By using different biochemical conjugations of the dye, the phagosome can be delineated from the cytoplasm so that changes in the size and shape of the phagosome can be assessed. This allows for further monitoring of ionic movement.

Distribution of translation initiation factor eIF3 in neutrophils in Alzheimer disease

The distribution of initiation factor eIF3 in the cytoplasm of peripheral blood neutrophils in Alzheimer disease (AD) was investigated. Using a specially developed image analysis protocol, we found a significant increase in the heterogeneity of the eIF3 distribution in AD neutrophils as compared with control cells. We also obtained an evidence that the increase in the eIF3 heterogeneity in AD neutrophils is associated with the formation of stress granules–non-membrane structures that emerge in the cytoplasm of eukaryotic cells in response to the environmental stress.

Prognostic significance of C5a2 on polymorphonuclear neutrophil and C5a2intra/C5a2 ratio level for early sepsis in an ED

IntroductionThe current study was designed to evaluate the expression of the second C5a receptor (C5a2) on polymorphonuclear neutrophil and in the cytoplasm of polymorphonuclear neutrophils (C5a2intra) in patients with sepsis in the emergency department (ED) for risk stratification and mortality. MethodsConsecutive patients fulfilling the criteria for systemic inflammatory response syndrome (n = 357) were admitted to Beijing Chao-Yang Hospital ED between January 2015 and July 2015. They were enrolled to identify the expression of C5a2 and C5a2intra and categorized into the following 4 groups: systemic inflammatory response syndrome, sepsis, severe sepsis, and septic shock. ResultsWe report that the surface C5a2 decreased and the C5a2intra/C5a2 ratio level increased with sepsis severity. As independent predictors of 28-day mortality, the areas under the receiver operating characteristic curves of combination of C5a2 or C5a2intra/C5a2 ratio level and the Mortality in ED Sepsis score were significantly higher than that of procalcitonin alone in predicting 28-day mortality in septic patients. ConclusionThe C5a2 and the C5a2intra/C5a2 ratio levels are probably valuable for the risk stratification of sepsis and are associated with the mortality of early sepsis in the ED.

Primer Reporte de Hepatozoon procyonis en Mapaches de Costa Rica

El estudio muestra, por primera vez, la detección de Hepatozoon procyonis en prociónidos, Procyon lotor de Costa Rica. Para caracterizar los gametocitos, se tomó muestras de sangre de mapaches infectados naturalmente. Los gametocitos concuerdan con Hepatozoon procyonis, los cuales fueron observados en neutrófilos del citoplasma, resultando su morfología similar a la reportada en mapaches de todas las Américas.

The Thyroid Hormone Inactivating Enzyme Type 3 Deiodinase is Present in Bactericidal Granules and the Cytoplasm of Human Neutrophils.

Neutrophils are important effector cells of the innate immune system. Thyroid hormone (TH) is thought to play an important role in their function. Intracellular TH levels are regulated by the deiodinating enzymes. The TH-inactivating type 3 deiodinase (D3) is expressed in infiltrating murine neutrophils, and D3 knockout mice show impaired bacterial killing upon infection. This suggests that D3 plays an important role in the bacterial killing capacity of neutrophils. The mechanism behind this effect is unknown. We aimed to assess the presence of D3 in human neutrophils, and determine its subcellular localization using confocal and electron microscopy, because this could give important clues about its function in these cells. D3 appeared to be present in the cytoplasm and in myeloperoxidase containing azurophilic granules and as well as lactoferrin containing specific granules within human neutrophils. This subcellular localization did not change upon activation of the cells. D3 is observed intracellularly during neutrophil extracellular trap formation, followed by a reduction of D3 staining after release of the neutrophil extracellular traps into the extracellular space. At the transcriptional level, human neutrophils expressed additional essential elements of TH metabolism, including TH transporters and TH receptors. Here, we demonstrate the presence and subcellular location of D3 in human neutrophils for the first time and propose a model, in which D3 plays a role in the bacterial killing capacity of neutrophils either through generation of iodide for the myeloperoxidase system or through modulation of intracellular TH bioavailability.

Human Neutrophil Elastase Inhibitory Alkaloids from Chelidonium majus L.

Abstract Human neutrophil elastase (HNE) represents a goodtherapeutic target for the treatment of inflammatory diseases aswell as invasion of microorganism. The methanol extract of aaerial part of Chelidonium majus L. showed high activity againstthe neutrophil elastase with an IC 50 value of 100µg/mL. Due toits potency, subsequent bioactivity-guided fractionation ofmethanol extract led to six alkaloids (1-6), which were identifiedas dihydrosanguinarine (1), (s)-stylopine (2), arnottianamide (3),(+)-chelidonine (4), spallidamine (5), and N-trans-feruloyltyramine (6). Among of them, three alkaloids (2, 5, and 6)inhibited HNE in a dose-dependent manner with IC 50 rangingbetween 11.6 and 51.0µM. Lineweaver-Burk and Dixon plots,and their secondary replots showed that alkaloids (2, 5, and 6)were mixed inhibitors of HNE. The analysis of K I and K IS valueproved that all inhibitors (2, 5, and 6) had reversible mixed typeI mechanism.Keywords Chelidonium majus L. · human neutrophil elastase ·inflammation · isoquinoline alkaloidIntroductionThe regulation of the enzymatic activity of human neutrophilelastase (HNE) has been the attractive field because neutrophilsare the first cells recruited to inflammatory sites and from theearliest line of defense against the invasion of microorganism(Brice et al., 2010). The human neutrophil elastase (EC 3. 4. 21.37) is the family of serine proteases that possess the ability tohydrolyze the extracellular matrix protein (Siedle et al., 2007). Itis present in azurophil granules in the neutrophil cytoplasm. HNEis a proteolytic enzyme involved in pathogenesis of emphysema,adult respiratory distress syndrome and rheumatoid arthritis(Crocetti et al., 2013). The catalytic site of HNE molecule iscomposed of the triad His41-Asp99-Ser173, of which the oxygenof serine attacks carbonyl group on the target substrate (Bode etal., 1989). The activity of HNE is controlled by inhibitor namedα1-antitrypsin produced in the liver. However, their HNE affinityis strongly decreased by oxidative stress and by proteases releasedfrom leukocytes that are recruited to inflammation sites (Brice etal., 2010). Thus, the imbalance between HNE and its inhibitors isimplicated in many inflammation diseases as like pulmonaryemphysema.Chelidonium majus L. is a perennial herbaceous plant of thefamily Papaveraceae, which is widely distributed around theworld. This plant has been traditionally used as an herbal medicinefor treatment of gastric ulcer, oral infection and liver disease(Lenfield et al., 1981). These effects are mainly due to the alkaloidspresent in milk sap. The major bioactive components of this plantare isoquinoline alkaloids such as chelidonine, chelerythrine,sanguianarine, bebeerine and coptisine (Barreto et al., 2003). Itincluded flavonoids and various acids such as ferulic, cumaric,caffeic and chelidonic acids (Barens J et al., 2007). The constituentsof C. majus L. have been found to exhibit antitumor, antiviral, andanti-inflammatory activities (Colombo and Bosisio, 1996). Additionally,the potent and selective acetylcholinesterase inhibition wasobserved on 8-hydroxydihydrochelerythrine in C. majus L (Cho etal., 2006).During a screening procedure on higher plant to find neutrophilelastase inhibitors, methanol extract of C. majus L. was shown toexhibit considerable inhibitory activity. In this study, we isolatedsix alkaloids targeting to HNE from the methanol extract of aerial

Sox2 functions as a sequence-specific DNA sensor in neutrophils to initiate innate immunity against microbial infection

Neutrophils express Toll-like receptors (TLRs) for the recognition of conserved bacterial elements to initiate antimicrobial responses. However, whether other cytosolic DNA sensors are expressed by neutrophils remains elusive. Here we found constitutive expression of the transcription factor Sox2 in the cytoplasm of mouse and human neutrophils. Neutrophil-specific Sox2 deficiency exacerbated bacterial infection. Sox2 directly recognized microbial DNA through its high-mobility-group (HMG) domain. Upon challenge with bacterial DNA, Sox2 dimerization was needed to activate a complex of the kinase TAK1 and its binding partner TAB2, which led to activation of the transcription factors NF-κB and AP-1 in neutrophils. Deficiency in TAK1 or TAB2 impaired Sox2-mediated antibacterial immunity. Overall, we reveal a previously unrecognized role for Sox2 as a cytosolic sequence-specific DNA sensor in neutrophils, which might provide potential therapeutic strategies for the treatment of infectious diseases.

Localization of annexins A1 and A2 in the respiratory tract of healthy calves and those experimentally infected with Mannheimia haemolytica.

Annexins A1 and A2 are proteins known to function in the stress response, dampening inflammatory responses and mediating fibrinolysis. We found, in healthy cattle recently arrived to a feedlot, that lower levels of these proteins correlated with later development of pneumonia. Here we determine the localization of annexin A1 and A2 proteins in the respiratory tract and in leukocytes, in healthy calves and those with Mannheimia haemolytica pneumonia. In healthy calves, immunohistochemistry revealed cytoplasmic expression of annexin A1 in the surface epithelium of large airways, tracheobronchial glands and goblet cells, to a lesser degree in small airways, but not in alveolar epithelium. Immunocytochemistry labeled annexin A1 in the cytoplasm of neutrophils from blood and bronchoalveolar lavage fluid, while minimal surface expression was detected by flow cytometry in monocytes, macrophages and lymphocytes. Annexin A2 expression was detected in surface epithelium of small airways, some mucosal lymphocytes, and endothelium, with weak expression in large airways, tracheobronchial glands and alveolar septa. For both proteins, the level of expression was similar in tissues collected five days after intrabronchial challenge with M. haemolytica compared to that from sham-inoculated calves. Annexins A1 and A2 were both detected in leukocytes around foci of coagulative necrosis, and in necrotic cells in the center of these foci, as well as in areas outlined above. Thus, annexins A1 and A2 are proteins produced by airway epithelial cells that may prevent inflammation in the healthy lung and be relevant to development of pneumonia in stressed cattle.Electronic supplementary materialThe online version of this article (doi:10.1186/s13567-014-0134-3) contains supplementary material, which is available to authorized users.

Neutrophils Promote Hematopoiesis Via a Novel Mechanism Involving Leucine Rich α-2 Glycoprotein (LRG)

Leucine-rich α2 glycoprotein (LRG), the founding member of the leucine-rich repeat superfamily of proteins, was initially identified in serum more than 30 years ago, but its biologic function has remained elusive. A role for LRG has been implicated in inflammation and angiogenesis. Our laboratory previously identified cDNA and genomic clones for human and murine LRG, and showed that ectopically expressed LRG localizes to the granule compartment in transfected myeloid cell lines and promotes their granulocytic differentiation. We also demonstrated that expression of LRG is transcriptionally regulated during neutrophil granulocyte differentiation in a manner similar to that reported for genes encoding the different subsets of neutrophil granule proteins. The presence of LRG in primary neutrophils and a role for LRG in hematopoiesis, however, have not been previously described. Based on our prior studies in transfected myeloid cell lines, we considered the tantalizing possibility that LRG is a novel neutrophil granule protein that is secreted extracellularly upon neutrophil activation to modulate hematopoiesis. To investigate this, we examined LRG in primary human neutrophils isolated from healthy volunteers. Immunoblot analysis of whole cell lysates from neutrophils (97% purity) identified a higher molecular weight LRG species in neutrophils (62 kDa) compared to serum (50 kDa); our data demonstrate the difference in apparent molecular weight is due to differential glycosylation. Immunofluorescence microscopy using antibodies to human LRG and antibodies to the neutrophil granule proteins myeloperoxidase (MPO), lactoferrin (LF), and matrix metalloproteinase 9 (MMP-9, also known as gelatinase), along with fluorescently-labeled secondary antibodies, demonstrated the presence of LRG in the cytoplasm of neutrophils in a compartment corresponding to LF. ELISA and immunoblot analyses of subcellular fractions from isolated neutrophils prepared by nitrogen cavitation demonstrated the presence of LRG in LF-containing fractions as well as some MMP-9-containing fractions, consistent with localization of LRG to the secondary/tertiary granule compartment. Neutrophil exocytosis assays using ionomycin, phorbol-12-myristate 13-acetate, and f-Met-Leu-Phe as stimulants also indicated that LRG is co-released with LF and MMP9, but not with MPO. Notably, LRG secreted from activated neutrophils could bind cytochrome c as reported for LRG purified from serum. Recent reports that LRG can also bind to the TGFβR1 receptor on endothelial cells prompted us to investigate the effects of LRG on TGFβ signaling in hematopoietic cells. LRG significantly antagonized the inhibitory effect of TGFβ on HL-60 cell proliferation (n=3; p<0.05) and also on colony growth of human hematopoietic progenitor cells. When LRG was added to hematopoietic progenitor cells cultured in TGFβ-containing Methocult (SF H4436, serum free), a 50% increase in CFU-GMs was observed. Collectively, these data suggest a novel mechanism whereby neutrophils modulate hematopoiesis in the microenvironment via extracellular release of LRG, and invoke an additional role for neutrophils in innate immunity that has not previously been reported DisclosuresNo relevant conflicts of interest to declare.

P-036 YI Clinical and Serological Factors Associated With Response to Infliximab in Chronic Refractory Pouchitis and Pouch Complications

Pouchitis refractory to first-line therapies and pouch complications remains problematic following colectomy and ileal-pouch anal anastomosis (IPAA) for ulcerative colitis (UC). Evidence for infliximab (IFX) use in this setting and factors predicting its success remain limited. Here, we investigated IFX use in refractory pouchitis and clinical and serological variables associated with treatment response. Patients were identified from the Mount Sinai Hospital IBD and Pelvic Pouch Databases, Toronto, Canada. Clinical, endoscopic and serological data were reviewed from 579 individuals who underwent colectomy and IPAA from 2000 to 2014. Patients with chronic refractory pouchitis and Crohn’s Disease- like outcome treated with IFX were included. Pretreatment parameters were measured within 4 weeks of induction and IFX response at a median of 9 (initial) and 48 weeks (sustained) respectively. Complete response was defined as symptomatic and endoscopic resolution with modified Pouchitis Disease Activity Index (mPDAI) score <5. Partial response was defined as improvement in symptoms with reduction in mPDAI >2. Endoscopic mucosal healing, pouch excision, pouch complications and CRP were recorded. Serum was analyzed for antibodies against Saccharomyces cerevisiae (ASCA), OmpC, CBir1 and perinuclear neutrophil cytoplasm (pANCA). Fisher’s exact testing and Kruskal-Wallis detected differences in clinical and serological variables between subgroups. Logistic regression models were applied to estimate odds ratio (OR) and 95% confidence interval (CI) of clinical and serological factors with initial and sustained clinical response as dependent variables. Results were deemed significant if P-value was ≤0.05. Thirty-one patients were included (33% male; age 32.6 ± 2.6 [mean ± SE]). 26% were on combined therapy with immune-modulator. Seventy-four present achieved post-induction response at median 9 weeks (IQR [6.5–13]) (48% complete, with concomitant mucosal healing). At median 48 weeks (IQR [25–71.5]), 62.6% retained response (29.6% complete). There were significant reductions in mPDAI in the entire cohort from a pre-induction score of 8.5 ± 0.3 to 2 ± 3.4 at final follow-up (mean, SE; P < 0.002) and in CRP from 29.48 ± 6.2 mg/L to 5.76 ± 1.6 mg/L (P < 0.001). Age, gender, smoking, pre-pouch disease extent and CRP did not affect response to IFX. Pretreatment mPDAI score did not significantly affect sustained IFX response but higher pretreatment mPDAI scores were negatively associated with early mucosal healing (P = 0.056; OR = 0.5, 95% CI, [0.2–1.010]). Patients with pretreatment mPDAI >10 were less likely to have initial mucosal healing (P = 0.03). Initial mucosal healing was associated with sustained complete response (P < 0.05; [OR] = 13.2; 95% confidence interval [CI], 1.0–165). Presence of ASCA was associated with higher initial mPDAI (P = 0.016), but no difference in treatment response. IFX responders had a lower number of positive antibody titers (2 positive titers versus 3, mean; P < 0.05). Having >2 positive titers was associated with longer term non-response to IFX though not significantly. IFX was effective in the short- and longer term in patients with chronic refractory pouchitis and pouch complications. Mucosal healing after induction is the strongest clinical association with a complete sustained response to IFX in this group and pretreatment mPDAI may help predict this.

Differential expression of pentraxin 3 in neutrophils

Pentraxins belong to the superfamily of conserved proteins that are characterized by a cyclic multimeric structure. Pentraxin 3 (PTX3) is a long pentraxin which can be produced by different cell types upon exposure to various inflammatory signals. Inside the neutrophil PTX3 is stored in form of granules localized in the cytoplasm. Neutrophilic granules are divided into three types: azurophilic (primary) granules, specific (secondary) granules and gelatinase (tertiary) granules. PTX3 has been considered to be localized in specific (secondary) granules. Immunofluorescent analyses using confocal laser microscopic examination were performed to clarify the localization of all three groups of granules within the cytoplasm of the mature neutrophils and neutrophils stimulated with IL-8. Furthermore, PTX3 was localized in primary granules of promyelocyte cell line HL-60. As a result, we suggest that PTX3 is localized not only in specific granules, but is also partly expressed in primary and tertiary granules. After the stimulation with IL-8, irregular reticular structures called neutrophil extracellular traps (NETs) were formed, three types of granules were trapped by NETs and PTX3 showed partial colocalization with these granular components. PTX3 localized in all three types of granules in neutrophils may play important roles in host defense.

Cytomorphological studies on granulogytes of pig

Author for correspondence : SURESH MEHTA Department of Veterinary Anatomy, Ranchi Veterinary College, RANCHI (JHARKHAND) INDIA Email: drsureshmehtakavaya@gmail.com Abstract : The neutrophils were round in shape and 12.00±0.44 mm in size. The nucleus possessed lobes with variable size, shape and number. The pale gray cytoplasm of neutrophils was laden with light pinkish fine granules which were uniformly distributed. The eosinophils were round in shape and were 12.25±0.28 mm in size. The nuclei had 2-4 lobes and connected with thick chromatin strands The cytoplasmic granules were round, numerous, densely packed, strongly eosinophilic and uniformly distributed throughout the cytoplasm. The basophils were roughly round in shape and 13.25±0.24 mm in size. The nuclei had 2-3 lobes which were connected with comparatively thicker chromatin strand. The cytoplasm had numerous basophilic granules occupying whole of the cytoplasm. The erythrocytes of pig were non nucleated, biconcave shaped and 5.7±0.17 mm in size. The platelets were irregular in shape and 2.76±0.33 mm in size.

P-057A: Detecting inflammation in pediatric inflammatory bowel disease: the role of fecal calprotectin and its impact on management

Introduction: Fecal calprotectin (FC) is a calcium binding protein found in the cytoplasm of neutrophils, monocytes and macrophages. FC levels can be used as a marker of inflammation in inflammatory bowel disease (IBD) patients. Aim: The aim of this study is to explore the current use of FC at our pediatric hospital, and to assess the impact of FC results on patient management. Methods: We performed a retrospective chart review of all IBD patients who had an FC level checked between January 2012 and January 2014. Demographic information, laboratory values and the indications for checking FC were collected. We documented any change in patient management that occurred as a result of the FC value. Results: Forty-seven patients were included in our review. Thirty-seven of the patients (79%) were found to have FC levels greater than 200 mg/g indicating active ongoing inflammation. Thirteen of thirty-seven patients (35%) with elevated FC levels had normal routine blood work screening (including erythrocyte sedimentation rate and c-reactive protein). The FC level resulted in management changes in 32/47 patients (68%). Conclusion: FC was a valuable screening test for inflammation in children with IBD in our center. FC detected inflammation in a subgroup of patients with otherwise normal screening inflammatory markers. FC also provided new information to the physician that frequently resulted in a change in management.