Double negative alpha beta T cells in pediatric hemophagocytic syndromes

Introduction: Autoimmune lymphoproliferative syndrome (ALPS) and hemophagocytic lymphohistiocytosis (HLH) share clinical and laboratory features including lymphadenopathy, splenomegaly, and pancytopenia. We sought to measure αβ double negative T cells (αβ DNT) in a group of patients with established diagnosis of HLH in relation to disease activity and severity.Methods: We conducted a follow-up, controlled study that comprised 25 patients with HLH and 25 healthy matched controls. Patients were subjected to clinical evaluation and flowcytometric measurement of αβ DNT Cells at presentation and 9 weeks after start of HLH induction treatment.Results: In 17 (68%) patients, infection was the trigger of HLH while the cause was malignancy in three (12%), and rheumatological disorders in two patients (8%). At enrollment, 15 patients (60%) had αβ DNT cells levels [median (IQR): 1.71 (1.25-2.12)] that were significantly higher than the control values [median (IQR): 0.7 (0.4-0.8)] (p<0.001). The αβ DNT counts of patients were also higher at enrollment as compared to values at the end of week 9 [median (IQR): 0.76 (0.45-1.17)]; p=0.018. Survivors (n=8) and non-survivors (n=17) had comparable levels of αβ DNT cells at enrollment (p=0.861). αβ DNT cell count correlated positively with ALT (p=0.019) and negatively with CD4/CD8 ratios (p=0.023).Conclusion: Elevated αβ DNT cell counts might be related to the HLH process and this implies that mild elevation can exist in HLH and are not specific to ALPS. Wider scale studies with longer periods of follow up are needed to validate the results and properly outline the correlation between both medical conditions.Keywords: Hemophagocytic lymphohistiocytosis, Double negative T cells, mortality, ALP

ISOLATION AND IDENTIFICATION OF Spiroplasma citri USING SOME SENSITIVE METHODS

Spiroplasma citri, the causal agent of stubborn disease, was isolated from leaves and fruits of diseased citrus plants and cultured on solid and liquid C3G medium. On the basis of mode of transmission, symptomatology, shape on solid medium, staining of infected plant tissues with Dienes’ stain and phase contrast microscopy, the isolated agent was identified as Spiroplasma citri. Identifiction was ensured by ELISA and PCR techniques. A fragment (1053 bp) from the spiralin gene of S. citri was amplified by PCR using two specific primers for the spiralin gene. The nucleic acid hybridization techniques (Southern and dot blots) were used for identification of the spiroplasmal genome using non-radioactive DNA probe specific for spiralin gene region. It was carried out as an alternative sensitive method for rapid detection of the Egyption isolate of S. citri   using non-radioactive DNA probe specific for spiralin gene region

Comparative Evaluation for the Effect of Green tea, Aloe vera and Chlorhexidine on Dentin Erosion

ABSTRACT: Purpose: This study aimed to compare the anti-collagenolytic effect of Green tea, Aloe vera and chlorhexidine on artificial erosion in human dentin using microhardness test and environmental scanning electron microscope (ESEM). Materials and Methods: For this study, 60 extracted sound human maxillary premolars were subjected to artificial erosion and randomly divided into three equal groups (20 premolars each) according to immersion in the tested anti-collagenolytic agents; group (A1) Green tea, (A2) Aloe vera and group (A3) Chlorhexidine (CHX). The microhardness analysis and (ESEM) photomicrographs were taken for the surface of dentin specimens three times, at base line, after the artificial erosion and after immersion in each treatment solution. Bonferroni’s post-hoc test was used for pair-wise comparisons. Statistical analysis was performed with IBM SPSS Statistics for Windows. Results: Results showed no statistically significant difference in microhardness mean values between Aloe vera and (CHX), However, both showed statistically significant increase in microhardness values than Green tea. These results were also supported by (ESEM) photomicrographs. Conclusion: Chemical gold standard CHX could be substituted by natural safe herbal sources and hence Aloe vera opens a new perspective in management of dental erosion

Influence of Etching Methods on the Shear Bond Strength of a Universal Adhesive System to Sound and Artificial Caries-Affected Dentin

Abstract: This study tested the effect of different etching methods on the bond strength of a universal adhesive system to caries-affected (CAD)and sound dentin. Methods: forty premolars were wet grinded to expose flat occlusal midcoronal dentin. Specimens were equally divided according to the substrate; sound and (CAD) by exposing to pH-cycling for 14 days. Each dentin substrate was bonded with Single BondTM universal adhesive applied either after no etching or etching of dentin, then resin composite was built up. Specimens were tested for shear bond strength (SBS). The data was analyzed by two-ways ANOVA. Results: Single BondTM universal bonded to sound dentin had statistically significant higher SBS mean values compared to CAD. Single BondTM applied after separate etching step of dentin showed a statistically higher SBS compared to self-etching method when bonded to sound dentin. Conclusion: Single BondTM applied after dentin separate etching step improved only bonding to sound dentin. CAD had a negative influence on the SBS of the tested universal adhesive

Evaluation of asymptomatic venous disease by Doppler ultrasonography in Behçet’s disease patients

Aim of the work: Evaluation of asymptomatic venous disease in patients with Behçet’s disease (BD) using venous Doppler ultrasonography (US) and its relation to different disease manifestations and activity. Patients and methods: Twenty-two BD patients (20 males and 2 females) with a mean age of 36.9 ± 10.6 years and disease duration of 10.8 ± 11.3 years without any known vascular disease and 22 age and sex matched controls were enrolled in this study. The Behçet’s disease current activity form (BDCAF) was used to assess disease activity. Patients and controls were subjected to venous Doppler US for both upper and lower limbs as well as the inferior vena cavae (IVC). The Clinical-Etiology-Anatomy-Pathophysiology (CEAP) severity score was used to evaluate the severity of venous insufficiency. Results: The 22 patients had a mean BDCAF score of 2.2 ± 0.2. No venous thrombosis or thrombophlebitis was detected in patients or controls. Three patient (13.6%) and one control (4.5%) revealed venous-insufficiency (venous reflux >1 s) in the lower extremities. The venous-insufficiency involved the superficial venous system and involved the deep venous system in 1 patients and the control. The lower limb veins were normal on both sides as regards compressibility, wall thickness and competency of perforator veins. Upper extremity veins were totally normal in all subjects. Conclusion: No superficial, deep venous dysfunction on both lower or upper limbs and/or IVC thrombosis was found in BD patients. Further follow-up venous Doppler for BD patients even without vascular complications is recommended to detect subclinical cases that may predict future thrombotic events

Outcome of different treatment regimens used in newly diagnosed ITP pediatric patients

Aim of the study: To compare the efficacy of different treatment options for newly diagnosed ITP in pediatrics. Patients and methods: This is a retrospective observational study that reviewed data of 102 newly diagnosed ITP children. Treatment regimens received by patients were classified into 5 types: Type I: pulse intravenous methylprednisolone followed by oral prednisolone then gradual tapering. Type II: oral prednisolone 4 mg/kg/d × 4 days. Type III: oral prednisolone 2 mg/kg/d followed by gradual tapering. Type IV: intravenous immune globulin (IVIG) combined with intravenous methylprednisolone. Type V: no treatment. Platelet counts at presentation, after 1 week, at 1 month, 3 months and 6 months from initial presentation were analyzed. Results: Patients who received Type I, III or IV treatment showed statistically significant higher response to treatment at 1 week (96.1%, 93.3%, and 100% respectively) in comparison to those who received Type II or no treatment (Type V) in which less response was observed (58.8% and 75% respectively); (p-value = 0.001%). However, the overall response showed no statistically significant difference in response to treatment using different types of treatment at one month and 3 months, p-value = 0.086 and 0.335 respectively. When we evaluated the response overtime of each line of treatment at 3 months, Type I showed a statistically significant response (p = 0.009) with the biggest number of patients continuing to maintain response up to 3 months. Conclusion: We recommend the use of (methylprednisolone 20–30 mg/kg/d × 3–5 days) followed by oral (prednisolone 2 mg/kg/d) due to rapid and sustained response. Keywords: Purpura, Thrombocytopenia, Idiopathic, Methylprednisolone, Immunoglobulins, Intravenous, Prednisolon

Improvement of DAS-28 ESR score in Egyptian children and adolescents with juvenile idiopathic arthritis treated with etanercept

Aim of study: To assess the efficacy and safety of using anti-tumor necrosis factor-α (TNF-α), Etanercept (ETN) in treatment of children and adolescents with Juvenile Idiopathic arthritis (JIA). Patients and methods: This retrospective cohort study included 18 children and adolescents who were diagnosed as JIA and treated with anti-tumor necrosis factor-α (TNF-α), ETN for at least 6 months due to failure of synthetic disease-modifying antirheumatic drugs (sDMARDS) as methotrexate (MTX). Before starting ETN treatment, all patients were receiving MTX in addition to non-steroidal anti-inflammatory drugs (NSAIDs). Other medications used were Prednisolone, Leflunamide and Hydroxychloroquine. As per recommendations of the European League Against Rheumatism (EULAR), failure of these DMARDs is an indication to start biological DMARDs. Anti TNF-α, mainly ETN is one of the most commonly used biological DMARDs in treating JIA. Clinical scoring using DAS28-ESR score was done for all patients at time of enrollment, 3 months and 6 months after starting ETN. The effect of different clinical and laboratory variables on the outcome of ETN treatment was assessed. Results: There was statistically significant improvement in DAS28-ESR after 3 months of treatment (p = 0.001) and after 6 months (p = 0.049). There was no statistically significant difference in DAS-28 score between males and females, patients younger than 10 years and patients older than 10 years, neither at onset nor at 6 months’ follow up. Similarly, the duration of illness before starting ETN did not affect DAS28 score at 6 months of treatment. Conclusion: ETN treatment of JIA children and adolescents demonstrated safety and efficacy in clinical improvement of DAS28-ESR score at 3 and 6 months of treatment. However, further studies on a bigger number of patients, longer duration and using different biological agents are needed

Cell Growth Inhibition, DNA Fragmentation and Apoptosis-Inducing Properties of Household-Processed Leaves and Seeds of Fenugreek (<i>Trigonella Foenum-Graecum</i> Linn.) against HepG2, HCT-116, and MCF-7 Cancerous Cell Lines

Household processing of fenugreek seeds and leaves, including soaking, germination, and boiling of the seeds, and air-drying of the leaves, has improved the levels of human consumption of the bitter seeds and increased the shelf life of fresh leaves, respectively. The potential anticancer activity of either unprocessed or processed fenugreek seeds or leaves and the relative expression of pro-apoptotic and anti-apoptotic genes of the studied cancerous cell lines exposed to IC50 crude extracts was investigated to observe the apoptotic-inducing property of this plant as an anticancer agent. The protein expression of IKK-α and IKK-β, as inhibitors of NF-KB which exhibit a critical function in the regulation of genes involved in chronic inflammatory disorders, were studied in the tested cancerous cell lines. In this study, the anticancer activity of household-processed fenugreek leaves and seeds against HepG2, HCT-116, MCF-7, and VERO cell lines was measured using an MTT assay. DNA fragmentation of both HepG2 and MCF-7 was investigated by using gel electrophoresis. RT-PCR was used to evaluate the relative expression of each p53, caspase-3, Bax, and Bcl-2 genes, whereas ELISA assay determined the expression of caspase-3, TNF-α, and 8-OHDG genes. Western blotting analyzed the protein-expressing levels of IKK-α and IKK-β proteins in each studied cell line. Data showed that at 500 µg mL−1, ADFL had the highest cytotoxicity against the HepG2 and HCT-116 cell lines. Although, each UFS and GFS sample had a more inhibitory effect on MCF-7 cells than ADFL. Gel electrophoresis demonstrated that the IC50 of each ADFL, UFS, and GFS sample induced DNA fragmentation in HepG2 and MCF-7, contrary to untreated cell lines. Gene expression using RT-PCR showed that IC50 doses of each sample induced apoptosis through the up-regulation of the p53, caspase-3, and Bax genes and the down-regulation of the Bcl-2 gene in each studied cell line. The relative expression of TNF-α, 8-OHDG, and caspase-3 genes of each HepG2 and MCF-7 cell line using ELISA assays demonstrated that ADFL, UFS, and GFS samples reduced the expression of TNF-α and 8-OHDG genes but increased the expression of the caspase-3 gene. Protein-expressing levels of IKK-α and IKK-β proteins in each studied cell line, determined using Western blotting, indicated that household treatments decreased IKK-α expression compared to the UFS sample. Moreover, the ADFL and SFS samples had the most activity in the IKK-β expression levels. Among all studied samples, air-dried fenugreek leaves and unprocessed and germinated fenugreek seeds had the most anti-proliferative and apoptotic-inducing properties against human HepG2, MCF-7, and HCT-116 cell lines, as compared to the VERO cell line. So, these crude extracts can be used in the future for developing new effective natural drugs for the treatment of hepatocellular, breast, and colon carcinomas