Use of CBCT to investigate the root canal morphology of mandibular incisors

Purpose

To investigate the root canal morphology of mandibular incisors using cone-beam computed tomography (CBCT).

Materials and methods

A total of 353 patients with 1,412 healthy, well-developed mandibular incisors were enrolled. Radiographic examination by CBCT was conducted as part of their routine examination, diagnosis and treatment planning. The following observations were made using CBCT: (1) the number of roots; (2) the number of canals; (3) canal configuration according to Vertucci’s classification; (4) the position of root canal bifurcations.

Results

Two canals were found in 10.9 % of mandibular central incisors, 25.5 % of lateral incisors and in 18.2 % of all the 1,412 mandibular incisors. Significantly, more lateral incisors possessed two canals than central incisors (p < 0.05). Of the teeth with two canals, type III incisors were the most prevalent, followed by types II, IV and V. Furthermore, 37.7 % of teeth were found to have root canal bifurcations that were at or near to the cortical-middle third junction regions of the roots.

Conclusion

Routine mode CBCT imaging was clinically useful for detection of two canals and determines the position of root canal bifurcations in mandibular incisors.     

Evaluation of the nasopalatine canal and variations with cone-beam computed tomography

Purpose

Although nasopalatine canal (NPC) is one of the most important anatomical structures in premaxillary region, few documents are available with regard to anatomic variations, size, and morphology of NPC in the literature. In recent years, the need for radiological identification of anatomical structures has grown with increased dental implant applications in anterior maxillary region with higher esthetic expectations. This study aimed at investigating the NPC’s shape and anatomical variations by cone-beam computed tomography (CBCT) in the Turkish society, in a particular region.

Methods

A total of 500 individual CBCT images were investigated in terms of morphological, dimensional, and anatomical features of NPC. In addition, the width of incisive foramen (IF) and foramina of Stenson were evaluated.

Results

There was no statistically significant difference (p = 0.234) between women and men in terms of NPC shape. Morphological assessment of NPC revealed that 38.78 % of NPCs were hourglass-like in shape, 27.35 % of canals were funnel-shaped, 9.18 % of canals were conical shaped, and 8.25 % of canals were cylindrical shaped. The mean IF width and NPC length was found as 5.06 and 12.59 mm, respectively. And the mean canal length was found significantly longer in men than women (p < 0.001).

Conclusion

It has been shown that NPC has a lot of anatomical variations with regard to its dimensions as well as its morphological appearance. Therefore, identification of the anatomical structure of this region with two- and three-dimensional imaging techniques maybe thought to be important in facilitating surgical management and preventing possible complications.     

Evaluation of location and dimensions of mandibular lingual foramina using cone-beam computed tomography

Objectives

The aim of this study was to assess the regional frequency and anatomical properties of mandibular lingual foramina (MLF) and their bony canals with cone-beam computed tomography (CBCT).

Materials and methods

A retrospective study was conducted by selecting images of the mandible from CBCT examination of 500 patients. MLF were located according to tooth areas and were grouped into midline, paramedian, and posterior foramina. In addition, the frequency of bony canals originating from lingual foramina was calculated, and the course and anastomoses were examined.

Results

In total, 491 areas with lingual foramina were observed. The highest regional frequency was recognized in the midline area (95.2 %), followed by left first premolar (15.1 %) area. The frequency of foramina in the midline was different from the paramedian and posterior (p < 0.01) regions. 95.6 % of lingual vascular canals originating from midline lingual foramina had a perpendicular course into the symphysis, whereas 60.3 % of canals from paramedian foramina and 83.6 % of canals from lateral lingual foramina presented with an anteriorly directed course.

Conclusions

CBCT examination easily demonstrates the presence of the lingual vascular canals. MLF are frequently present in a Turkish population; radiologists and oral surgeons should be aware of this anatomic feature and its possible implications.     

Lateral lingual vascular canals of the mandible: a CBCT study of 500 cases

Purpose

The aim of the present study was to use cone-beam computed tomography (CBCT) images of patients to assess the localization, diameter, and course of the lateral lingual vascular canal (LLVC).

Methods

CBCT (Morita, Accuitomo 170) images from 500 patients were used in the present study. The CBCT images were examined by two oral radiologists. The diameter, localization, and course of the LLVCs were assessed using axial, cross-sectional, and multiplanar reformatted images. Statistical analysis was performed using SPSS^® v. 15 (SPSS Inc., Chicago, IL, USA), and t tests were used for statistical analysis.

Results

Of the 500 patients examined, 163 LLVCs were detected in 124 (24.8 %) cases. The mean diameter of LLVCs on the right side and left side was 0.65 ± 0.18 and 0.64 ± 0.17 mm, respectively. The majority of the LLVCs (n = 106, 83 %) were observed in the premolar region. There was a statistically significant difference between the occurrence of LLVCs in the premolar region and the other areas (p < 0.01).

Conclusions

All the LLVCs observed in this study had a connection with inferior alveolar canal structures. The detection of an LLVC on CBCT images should alert the radiologist and surgeons to the possible presence of collateral arterial and vascular structures.     

Presurgical CBCT assessment of maxillary neurovascularization in relation to maxillary sinus augmentation procedures and posterior implant placement

Purpose

To provide more information to clinicians planning sinus grafting and maxillofacial surgical interventions, the present study evaluated the prevalence, diameter and location of the superior alveolar canals (SAC) using CBCT images.

Methods

The maxillary sinus CBCT scans (i-CAT Classic^®, ISI, USA) of 100 adult patients (67 women and 33 men) aged 20–79 years [mean (SD) 40 (15)] were examined. A dentomaxillofacial radiologist observed the SAC based on CBCT image data and more specifically the parasagittal views to assess SAC’s diameter and location.

Results

The anterior and posterior SAC, double ASAC, intraosseous anastomoses and the extension of the anterior SAC to the piriform aperture were observed in 100, 73, 24.5, 38.5 and 84 % of the cases, respectively. The anastomosis was located between canine and first premolar in 43 % of the cases. The SAC diameters were in 80 % of the cases ≤1 mm, remaining canals had a diameter between 1 and 2 mm. The distance of the SAC to the alveolar crest ranged between 2.42 and 44.6 mm. The anterior SAC was more prevalent in the upper (53 %) and middle (44 %) thirds of the maxillary sinus, while the posterior SAC was more prevalent in the middle (36 %) and lower thirds (64 %). The distance was significantly bigger in men in some tooth positions.

Conclusions

Based on the present findings, one-fifth of the patients may have a diameter of the SAC >1 mm, large enough to cause bleeding and/or paraesthesia. CBCT imaging may assist surgeons to plan grafting and osteotomy procedures, while avoiding these neurovascular structures.     

Evaluation of the relationship between the maxillary posterior teeth and the sinus floor using cone-beam computed tomography

Aim

The aim of this study was to evaluate the relationship between each root of maxillary premolars and molars and the maxillary sinus floor according to sex, sinus position, and age by decade in a Turkish population by using cone-beam computed tomography (CBCT) scanning.

Methodology

We evaluated a database of 5,166 (2,680 maxillary premolars and 2,486 maxillary molars) CBCT scans obtained from 849 patients. The vertical relationship between each root of the molar and premolar teeth to the sinus floor was classified into three types: type 1, the roots penetrated into the sinus floor; type 2, the roots contacted the sinus floor; and type 3, the roots extended below the sinus floor.

Results

The results of the classification of each root in relationship to the sinus floor were as follows: type 3 occurred most frequently in the first (92.4 %) and second (71.6 %) premolar teeth, type 1 (34.2 %) occurred most frequently in the palatinal roots of the first molar teeth, type 3 occurred most frequently in the mesiobuccal (39.9 %) and distobuccal (39.7 %) roots of the first molar teeth, and type 2 (36.7 %) occurred most frequently in the mesiobuccal roots of the second molar teeth. No significant differences were found between the left and right sides, but several differences were found between males and females. The relationship between the posterior teeth and the sinus floor differed according to the age decade interval (p < 0.05).

Conclusions

The maxillary first premolars have no relationship with the maxillary sinus floor, but the maxillary second molars are closer to the sinus floor. Also the second decade and males were most susceptible to undesirable results.     

Optimisation of Omalizumab Dosage in Patients with Severe Persistent Allergic Asthma Using recoveryELISA

Background

The objective of this study was to investigate the suitability of recoveryELISA as a method to monitor treatment with therapeutic antibodies using the example of omalizumab.

Methods

The recoveryELISA is a newly developed immunoassay technology that measures three parameters in one test: the free level of antigen, the level of therapeutic antibody and the specific dose-response interaction which represents the actual activity of the drug. A retrospective and observational analysis was performed on 197 serum samples from 17 patients (13 ± 4 years of age) with severe persistent allergic asthma who received add-on treatment with omalizumab.

Results

The mean omalizumab serum level during antibody therapy was 59 ± 45 µg/mL; the kit’s upper detection limit of 140 µg/mL was exceeded in 27 samples. Antibody concentrations between 50 and 140 µg/mL were found in 64 samples. Independent of the omalizumab dosage, nearly all measurements were in a range of absolute saturation as regards the IgE binding rate. Almost complete binding of IgE with a recovery of added labelled IgE of <1 % was reached within a maximum of 11 days.

Conclusions

The biochemical activity of therapeutic antibodies can be examined by recoveryELISA and their residual activity can be determined. Thus, further individualisation of therapy with biologics is possible using this test which seems to be suitable to diminish side effects and reduce costs.     

Clinical Heterogeneity of Immunodysregulation,Polyendocrinopathy, Enteropathy,X-linked: Pulmonary Involvement as a Non-Classical Disease Manifestation

Purpose

IPEX (Immunodysregulation, Polyendocrinopathy, Enteropathy, X-linked) is a rare X-linked recessive life-threatening disorder characterized by autoimmunity and early death. Pulmonary complication related with IPEX has not been elucidated exactly. Here, we report 4 IPEX patients, 3 of which died from severe pulmonary disease.

Methods

Clinical data and laboratory findings including autoantibodies, immunoglobulin levels as well as number of T, B and NK cells were evaluated. FOXP3 expression and T reg activity were analyzed. The FOXP3 gene was sequenced and RNA analysis was performed.

Results

Patient I (PI) presented with nephrotic syndrome at 3 years of age and then developed autoimmune hepatitis without eczema, enteropathy or high IgE and died at 9 years of age due to acute respiratory distress syndrome (ARDS). Two cousins of PI had the same hypomorphic splice site mutation leading to a deletion of 27 amino acids, but normal FOXP3 protein expression and normal suppressive capacity of T reg in a proliferation inhibition assay. However, they exhibited typical symptoms such as eczema, diabetes and enteropathy with eosinophilia at early age (PII, PIII) and were transplanted in infancy. One of them had severe respiratory distress right after birth (PIII). Patient IV from another family presented with chronic diarrhea without autoimmune manifestations and died due to ARDS.

Conclusion

Lung disease related to IPEX syndrome has not been reported before and this entity could be a critical factor in disease outcome.     

Increased Prevalence of Gastrointestinal Viruses and Diminished Secretory Immunoglobulin a Levels in Antibody Deficiencies

Purpose

Gastrointestinal disease occurs frequently in antibody deficiencies. This study aims to explore the relation between gastrointestinal infections and mucosal homeostasis in patients with antibody deficiencies.

Methods

We performed an observational study including 54 pediatric antibody deficient patients (48 % CVID, 41 % CVID-like, 11 % XLA) and 66 healthy controls. Clinical symptom scores and stool samples were collected prospectively. Stool samples were evaluated for bacteria, parasites, viruses, secretory IgA- and for calprotectin levels. Results were compared between patients and controls.

Results

24 % of antibody deficient patients versus 9 % of healthy controls tested positive for gastrointestinal viruses (p?=?0.028). Fecal calprotectin levels were significantly higher in virus positive patients compared to virus negative patients (p?=?0.002). However, in controls, fecal calprotectin levels were similar between virus positive and virus negative controls. Moreover, gastrointestinal virus positive patients had low serum IgA levels in 13/14 cases (94 %) versus 40/62 (62 %) patients in the virus negative patient group (p?=?0.04). The virus positive patient group also displayed significantly lower secretory IgA levels in stool (median 13 ug/ml) than patients without gastrointestinal viruses detected or healthy controls (median 155 ug/ml) (p?=?0.046).

Conclusion

We here report an increased prevalence of gastrointestinal viruses and gastrointestinal complaints in antibody deficient patients. Patients that tested positive for gastrointestinal viruses showed diminished serum- and secretory IgA levels, and only in patients, virus positivity was associated with signs of mucosal inflammation. These findings suggest that particularly patients with low IgA are at risk for longstanding replication of gastrointestinal viruses, which may eventually result in CVID-related enteropathy.     

Mutational profiling of the RAS,PI3K,MET and b-catenin pathways in cancer of unknown primary: a retrospective study of the Hellenic Cooperative Oncology Group

Cancer of unknown primary origin (CUP) had a poor prognosis, determined by clinico-histological characteristics, partly due to the lack of insights on its biology. We screened tumour DNA from 87 patients with CUP for CTNNB1 (coding exons 2,3,4,5), MET (coding exon 18), PIK3CA (coding exons 9,20), KRAS (coding exons 1,2), BRAF (coding exon 15) gene mutations by using dd-sequencing and evaluated their impact on prognosis. Mutated gene incidences in the 87 CUP cases were: KRAS 11 (12.6 %), BRAF 5 (5.7 %), PIK3CA 8 (9 %), MET 6 (6.7 %) and CTNNB1 18 (20.7 %). Several mutations in the KRAS gene were not the commonly encountered mutations in other solid tumours. Activating mutations were observed in 10.2 % in KRAS, 4.5 % in BRAF, 6.6 % in PIK3CA, 4.5 % in MET, and 19.5 % in CTNNB1. Activating mutations in PIK3CA coding exon 9 were inversely correlated with MET coding exon 18 activating mutations (p = 0.036). MET activating mutations were prognostic for poor Progression-Free Survival (median PFS 5 vs 9 months, p = 0.009) and Overall Survival (median OS 7 vs 20 months, p = 0.005). The complex profile of either CTNNB1 or MET mutations also had an adverse prognostic significance (median OS 11 vs 21 months, p = 0.015). No other gene mutation exhibited prognostic significance. In multivariate analysis, poor performance status, male gender, visceral disease and adenocarcinoma histology, but not gene mutations, were independently associated with poor patient outcome. CTNNB1 gene mutations are frequent, and along with MET mutations have an adverse prognostic effect in patients with CUP.     

Terpinen-4-ol and alpha-terpineol (tea tree oil components) inhibit the production of IL-1β, IL-6 and IL-10 on human macrophages

Objective

Tea tree oil (TTO) is an essential oil with anti-inflammatory properties, steam distilled from the plant Melaleuca alternifolia. We investigated the immunomodulatory properties of TTO and its components (terpinen-4-ol and alpha-terpineol) using lipopolysaccharide (LPS)-stimulated macrophages.

Methods

The ability of TTO, terpinen-4-ol and alpha-terpineol to modulate the macrophage response to bacterial LPS stimulation was assessed by ELISA for tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and IL-10 cytokine production and by western blotting for the activation of nuclear factor kappa B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) signaling, which are associated with the expression of pro-inflammatory cytokines. We used a human monocytic cell line (U937) differentiated into macrophages.

Results

LPS induced the production of all cytokines, and TTO and its components significantly reduced the production of IL-1β, IL-6 and IL-10. The production of TNF-α was not affected by either TTO or its major components. The modulation of cytokine production was not mediated by changes in NF-κB or p38 MAPK activation.

Conclusion

TTO, terpinen-4-ol and alpha-terpineol can suppress the production of inflammatory mediators in LPS-stimulated human macrophages; this inhibition was mediated by interfering with the NF-kB, p38 or ERK MAPK pathways.     

Piceatannol inhibits phorbol ester-induced expression of COX-2 and iNOS in HR-1 hairless mouse skin by blocking the activation of NF-κB and AP-1

Objectives

The present study was aimed at elucidating the molecular mechanisms of anti-inflammatory activity of piceatannol (trans-3,4,3′,5′-tetrahydroxystilbene) in mouse skin in vivo.

Methods

Female HR-1 hairless mice were topically treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) with or without piceatannol pretreatment. Epidermal protein expression was assessed by Western blot analysis. The cyclooxygenase-2 (COX-2) expression was detected by immunohistochemistry. The DNA binding of nuclear factor-kappaB (NF-κB) and activator protein-1 (AP-1) was examined by the electrophoretic mobility gel shift assay. The catalytic activity of IκBα kinase-β (IKKβ) was measured by in vitro kinase assay.

Results

Pretreatment with piceatannol attenuated TPA-induced expression of COX-2 and inducible nitric oxide synthase (iNOS) in mouse skin. Piceatannol diminished nuclear translocation and the DNA binding of NF-κB through the blockade of phosphorylation and subsequent degradation of IκBα. Piceatannol attenuated the catalytic activity of IKKβ and inhibited the phosphorylation of mitogen-activated protein (MAP) kinases in TPA-treated mouse skin. In addition, piceatannol decreased TPA-induced expression of c-Fos and the DNA binding of AP-1.

Conclusion

Piceatannol inhibits TPA-induced COX-2 and iNOS expression by blocking the activation of NF-κB and AP-1 via suppression of the IKKβ activity and phosphorylation of MAP kinases, which provides a mechanistic basis of its anti-inflammatory effects in mouse skin.     

The First US Domestic Report of Disseminated Mycobacterium avium Complex and Anti-Interferon-γ Autoantibodies

Introduction

Anti-interferon-γ (IFNγ) autoantibodies have been associated with disseminated mycobacterial infections, mostly in patients from Southeast Asia.

Purpose

We studied an American-born, Caucasian female with M. avium complex infection of the subglottic mucosa and brain for underlying etiologies of infection.

Methods

Plasma was screened for anticytokine autoantibodies using a Luminex-based approach. The ability of patient plasma to block IFNγ-induced STAT1 phosphorylation in normal blood cells was evaluated by flow cytometry with intracellular staining. Plasma inhibition of IFNγ production and IFNγ-induced cytokines in normal and patient blood cells washed of autologous plasma was also evaluated.

Results

Patient plasma contained high-titer IgG anti-IFNγ autoantibodies, primarily of the IgG₁ subclass. Patient but not control plasma prevented IFNγ-induced STAT1 phosphorylation and expression of the IFNγ-inducible cytokines tumor necrosis factor (TNF) α and interleukin (IL)-12 in normal blood cells. Patient blood cells washed free of autologous plasma demonstrated normal IFNγ production and response.

Conclusions

Disseminated nontuberculous mycobacterial infections should always prompt immune evaluation. This first case of disseminated nontuberculous mycobacterial infection and anti-IFNγ autoantibodies in an American-born Caucasian suggests that anti-cytokine autoantibodies are not racially or regionally restricted.     

CT Screening for Pulmonary Pathology in Common Variable Immunodeficiency Disorders and the Correlation with Clinical and Immunological Parameters

Background

Pulmonary disease is common in patients with common variable immunodeficiency disorders (CVID) and involves infections, chronic airway disease and interstitial lung disease. Chronic pulmonary disease is associated with excess morbidity and early mortality and therefore early detection and monitoring of progression is essential.

Methods and Purpose

Thin slice CT scan and pulmonary function were used to determine the prevalence and spectrum of chronic (pre-clinical) pulmonary disease in adult CVID patients regardless of symptoms. CT Scans were scored for airway abnormalities (AD) and interstitial lung disease (ILD). Other CVID related complications and B and T lymphocyte subsets were analyzed to identify patients at risk for pulmonary disease.

Results

Significant pulmonary abnormalities were detected in 24 of the 47 patients (51 %) consisting of AD in 30 % and ILD in 34 % of cases. In only 7 (29 %) of these 24 patients pulmonary function test proved abnormal. The presence of AD was correlated to (recurrent) lower respiratory tract infections despite IgG therapy. The presence of ILD was correlated to autoimmune disease and a reduction in the numbers of CD4?+?T cells, naïve CD4?+?T cells, naïve CD8?+?T cells and memory B cells and lower IgG through levels over time.

Conclusion

Preclinical signs of AD and ILD are common in CVID patients despite Ig therapy and do not correlate to pulmonary function testing. Patients at risk for ILD might be identified by the presence of autoimmunity or a deranged T cell pattern. Larger studies are needed to confirm these findings and to determine thresholds for the T lymphocyte subsets.     

Renal function and Na−K-ATPase in rats after suprarenal ligation of inferior vena cava

To assess the effects of altered renal function on Na?K-ATPase, the following groups of rats were studied:

1. rats with suprarenal vena cava ligation (SVCL), la. DOCA-treated rats with SVCL,
2. rats with infrarenal vena cava ligation (IVCL),
3. rats with glycerol-induced acute renal failure,
4. rats with bilateral ureteric ligation, and
5. K-exalate-treated rats with SVCL.

In group 1, acute renal failure with hyperkalemia developed and medullary Na?K-ATPase increased from 95±5 in control to 155±7 μmol Pi/mg prot//h,P<0.001, DOCA did not prevent the increase of Na?K-ATPase. In group 2, medullary Na?K-ATPase decreased from 130±10 in control to 88±7,P<0.01, in rats with IVCL. In group 3, cortical Na?K-ATPase decreased from 55±5 to 27±6,P<0.02. In group 4, Na?K-ATPase was unchanged. In group 5, maintenance of normokalemia prevented the rise in Na?K-ATPase. These experiments demonstrated a K-dependent activation of medullary Na?K-ATPase after SVCL but not in other forms of renal failure. Because SVCL diminishes drastically GFR per nephron, the present findings imply that increased loads of Na and K per nephron are not a prerequisite for an increase in medullary Na?K-ATPase. Hyperkalemia in presence of increased renal venous pressure seems to be causually related to the rise in medullary Na?K-ATPase activity.     

Optimal Strategies to Identify Aberrant Intra-Epithelial Lymphocytes in Refractory Coeliac Disease

Introduction

Different strategies have been developed to identify those refractory celiac disease (RCD) patients who are at risk to develop an enteropathy associated T-cell lymphoma (EATL). Flow cytometric analysis of intra-epithelial lymphocytes (IEL) with an aberrant phenotype is considered the golden standard but is not widely available. Immunohistochemistry (IHC) and T-cell receptor (TCR) rearrangement studies are commonly available but may lack sensitivity and specificity. Here, we compared the three different methods in the workup of patients suspected for RCD.

Methods

Duodenal biopsies from control patient (n?=?5), RCD patients with moderately increased aberrant IEL populations (20–50 %: n?=?14), and RCD patients with high numbers of aberrant IEL (>50 %: n?=?5) as determined by flow cytometry were analysed by IHC and TCR-γ chain rearrangement analysis. Three pathologists scored the slides independently.

Results

Sensitivity of IHC and TCR-γ rearrangement analysis in RCD patients with high numbers of aberrant IELs was 100 and 71 %, respectively. RCD patients with aberrant cells between 25 and 50 % however, were missed by IHC and TCR in 50 and 57 % of cases, respectively. In addition, inter-rater reliability analysis of the IHC scoring revealed coder-pair Kappa coefficients between 0.28 and 0.85.

Conclusion

Immunohistochemistry and to a lesser extent TCR-γ clonality analysis are sensitive in identifying patients with high numbers of aberrant IEL populations, yet miss half of RCD patients with moderately increased numbers. In addition, IHC has a high inter-observer variability. Therefore, patients suspected for RCD should undergo flow cytometric analysis of the duodenum.     

Docosahexanoic acid antagonizes TNF-α-induced necroptosis by attenuating oxidative stress,ceramide production,lysosomal dysfunction,and autophagic features

Objective

It was previously reported that docosahexanoic acid (DHA) reduces TNF-α-induced necrosis in L929 cells. However, the mechanisms underlying this reduction have not been investigated. The present study was designed to investigate cellular and biochemical mechanisms underlying the attenuation of TNF-α-induced necroptosis by DHA in L929 cells.

Methods

L929 cells were pre-treated with DHA prior to exposure to TNF-α, zVAD, or Necrostatin-1 (Nec-1). Cell death and survival were assessed by MTT and caspase activity assays, and microscopic visualization. Reactive oxygen species (ROS) were measured by flow cytometry. C16- and C18-ceramides were measured by mass spectrometry. Lysosomal membrane permeabilization (LMP) was evaluated by fluorescence microscopy and flow cytometry using Acridine Orange. Cathepsin L activation was evaluated by immunoblotting and fluorescence microscopy. Autophagy was assessed by immunoblotting of LC3-II and Beclin.

Results

Exposure of L929 cells to TNF-α alone for 24 h induced necroptosis, as evidenced by the inhibition of cell death by Nec-1, absence of caspase-3 activity and Lamin B cleavage, and morphological analysis. DHA attenuated multiple biochemical events associated with TNF-α-induced necroptosis, including ROS generation, ceramide production, lysosomal dysfunction, cathepsin L activation, and autophagic features. DHA also attenuated zVAD-induced necroptosis but did not attenuate the enhanced apoptosis and necrosis induced by the combination of TNF-α with Actinomycin D or zVAD, respectively, suggesting that its protective effects might be limited by the strength of the cell death insult induced by TNF-α.

Conclusions

DHA effectively attenuates TNF-α-induced necroptosis and autophagy, most likely via its ability to inhibit TNF-α-induced sphingolipid metabolism and oxidative stress. These results highlight the role of this Omega-3 fatty acid in antagonizing inflammatory cell death.     

Sputum neutrophil count after bronchial allergen challenge is related to peripheral blood neutrophil chemotaxis in asthma patients

Objective

The aim of this study was to estimate relations between sputum neutrophilia and the chemotactic activity of peripheral blood neutrophils after the bronchial allergen challenge in asthma patients.

Materials and methods

Fifteen patients with allergic asthma (AA), 13 patients with allergic rhinitis (AR), all sensitized to Dermatophagoides pteronyssinus, and 8 healthy subjects (HS) underwent bronchial challenge with D. pteronyssinus. Sputum and peripheral blood collection were performed 24 h before, 7 and 24 h after the bronchial challenge. Cell counts were determined by the May-Grünwald-Giemsa method. Neutrophil chemotaxis was analyzed by a flow cytometer; IL-8 levels were measured by ELISA.

Results

Sputum neutrophil count and peripheral blood neutrophil chemotaxis of patients with AA were greater 7 and 24 h after the challenge compared with the baseline values and patients with AR and HS (P < 0.05). Moreover, a significant correlation was found between the neutrophil count in sputum and IL-8 levels, and the chemotactic activity of peripheral blood neutrophils 24 h after the bronchial challenge only the patients with AA (P < 0.05).

Conclusions

Increased sputum neutrophil count was found to be associated with an enhanced chemotactic activity of peripheral blood neutrophils during allergen-induced late-phase airway inflammation in patients with allergic asthma.