Antidiabetic and Liver Histological and Ultrastructural Effects of Cynara scolymus Leaf and Flower Head Hydroethanolic Extracts in Nicotinamide/Streptozotocin-Induced Diabetic Rats.

This study aims to investigate the effect of hydroethanolic extracts of Cynara scolymus (C. scolymus) leaf (CLHE) and C. scolymus flower (CFHE) on the hepatic histopathological lesions and functional biochemical changes induced by type 2 diabetes mellitus (T2DM). The rat model of T2DM was induced by intraperitoneal injection of streptozotocin (STZ) in a dose of 60 mg/kg for 15 minutes following nicotinamide (NA) (60 mg/kg). The rats were allocated into four groups: group 1 (negative control), group 2 (diabetic control), group 3 (diabetic rats supplemented with 100 mg/kg/day CLHE), and group 4 (diabetic rats supplemented with 100 mg/kg/day CFHE). Treatment with CLHE and CFHE, for the study duration of 28 days, significantly improved the deteriorated hepatic glycogen content, glycogen phosphorylase, glucose-6-phosphatase activities, serum fructosamine levels, lipid profile, aspartate transaminase activities, and alanine transaminase activities as well as serum insulin and C-peptide levels. The elevated liver lipid peroxidation and the decreased activities of superoxide dismutase and glutathione peroxidase were significantly alleviated. The elevated expression of the proinflammatory cytokine tumor necrosis factor-α in the liver of diabetic rats was significantly reduced by treatments with CLHE and CFHE. NA/STZ-induced T2DM exhibited hepatic histopathological changes in the form of disordered hepatocytes, cytoplasm dissolution, and mononuclear leukocytic infiltration. The electron microscopic ultrastructure study revealed damaged mitochondria with ill-defined cristae and fragmentation of the rough endoplasmic reticulum. Treatments with CLHE and CFHE remarkably amended these histopathological and EM ultrastructural changes. In conclusion, both CLHE and CFHE may have antidiabetic and improvement effects on the liver function and structural integrity, which may be mediated, at least in part, via suppression of inflammation and oxidative stress and enhancement of the antioxidant defence system.

Perfluorocarbon liquid-assisted vitreo-dissection in eyes with firmly adherent posterior hyaloid.

BACKGROUND: Induction of posterior vitreous detachment (PVD) is a critical step during pars plana vitrectomy. Multiple techniques and utilities have been proposed for assistance with this step with no consensus on the safest and most effective means, especially in eyes with firmly adherent posterior hyaloid. Viscodissection or the utilization of perfluorocarbon liquid (PFCL) can be used to dissect the posterior hyaloid and widely adherent epiretinal membranes. METHODS: A technique of PFCL dissection of the posterior hyaloid in eyes with abnormal adhesion of the posterior hyaloid. After core vitrectomy, breaking into the posterior hyaloid face is made via active aspiration and cutting or a sharp dissection. This is followed by active and slow injection of PFCL into the potential space between the posterior cortical vitreous and the neurosensory retina. A wave of PFCL propagates anteriorly causing "vitreo-dissection" of the peripheral cortical vitreous. RESULTS: The technique was effective and safe in 8 successive cases, 4 cases with vitreoretinal traction syndrome and 4 with diabetic tractional membranes. CONCLUSION: The technique can be considered in cases with abnormal firmly adherent posterior hyaloid when induction of PVD proves difficult.

Immune signature of CCR7+ central memory T cells associates with disease severity and Immunoglobulin E in bronchial asthma.

Summary: Objective. CD4+T cell subtypes are the central orchestrators of airway inflammation in bronchialasthma (BA); however, the mechanisms that regulate their accumulation in asthmatic airways are still a challenging subject. In addition, neutrophils play a significant role in the development of airway remodeling and their presence may influence clinical presentation of BA being linked to the development of severe BA. Neutrophils have also been found to acquireantigen presenting functions, enabling them to directly activate T cells. The study aimed to evaluate the possible association of chemokine receptor 7 (CCR7)+ memory CD4+T cells andCCR4+ effector T cells with disease severity and immunoglobulin E (IgE) production as well as to explore the relationship between these cells and neutrophil function in both allergic andnon-allergic asthmatic patients. Methods. Flow cytometry was used to determine the expression of different T cell subset phenotypes (CCR7 memory CD4+ and CCR4+T cells using anti-human CD3, CD4, CD45RO, CCR4 and CCR7 monoclonal antibodies) utilizing peripheral blood mononuclear cells (PBMCs) isolated from 78 allergic asthmatic patients, 41 non-allergic asthmatic patients, and 40 healthy individuals. Moreover, neutrophils' phagocytic activity was assessed by ingestion of candida particles. Results. We demonstrated increased percentages of CCR7+ memory CD4+T cells and CCR4+CD4+T cells in patients compared to control, where this up regulation was significantly higher in allergic than non-allergic asthmatic patients. Additionally, these cells were negatively correlated with improved pulmonary tests and significantly associated with disease severity scores and IgE levels. The neutrophil phagocytic activity was markedly increased in patients compared to control, showing a significant positive correlation with disease severity. Conclusions. These findings suggest that increased CCR4+ CD4+ T cells and CCR7+ memory CD4+ T cells (Tcm) may be associated with BA severity, especially in allergic BA patients and can potentially contribute to the rational design of new therapeutic approaches for asthma in the future.

Efficacy of Magnesium Sulfate Added to Local Anesthetic in a Transversus Abdominis Plane Block for Analgesia Following Total Abdominal Hysterectomy: A Randomized Trial.

BACKGROUND: Major abdominal surgeries are associated with severe pain, which can affect respiratory and cardiac functions if insufficiently treated; this increases postoperative morbidity. OBJECTIVE: We aim at evaluating the efficacy of magnesium sulfate as an adjuvant to local anesthetic in an ultrasound-guided transversus abdominis plane (TAP) block for postoperative analgesia in total abdominal hysterectomy. STUDY DESIGN: A prospective, randomized, double-blinded clinical trial. SETTING: An academic medical center. METHODS: This study is registered at https://clinicaltrials.gov (no.: NCT02930707). This randomized, double-blinded clinical trial included 60 women undergoing total abdominal hysterectomy that were divided into 2 groups (30 patients per group). Group I received a TAP block with 20 mL per side of 0.25% bupivacaine plus 2 mL magnesium sulphate 10% (200 mg). Group II received a TAP block with 20 mL per side of 0.25% bupivacaine. Visual analog scale (VAS) scores, the time of the first analgesic request, total morphine consumption, and any side effects were assessed and recorded. RESULTS: The mean postoperative VAS score was significantly reduced in group I compared to group II in all of the time-points except after 10 hours. The mean time of the first request for rescue analgesia was significantly prolonged in group I (15.67 hrs.) compared to group II (7.33 hrs.) (P < 0.001), and the mean total morphine consumption, over the first 24 hours postoperatively, was significantly lower in group I (7.63 ± 2.93 mg) than in group II (16.20 ± 3.24 mg) (P < 0.001). No significant difference in side effects was observed. LIMITATIONS: Sample size. CONCLUSION: The addition of 200 mg of magnesium sulfate to bupivacaine in an ultrasound-guided TAP block significantly reduced postoperative opioid requirements, prolonged the duration of analgesia, and reduced the VAS score in patients who underwent abdominal hysterectomy, without significant side effects. KEY WORDS: Magnesium sulfate, TAP block, postoperative pain, total abdominal hysterectomy.

An Approach to Breast Cancer Immunotherapy: The Apoptotic Activity of Recombinant Anti-Interleukin-6 Monoclonal Antibodies in Intact Tumour Microenvironment of Breast Carcinoma.

Current work is one of our comprehensive preclinical studies, a new approach to breast cancer (BC) immunotherapy through induction of tumour cell apoptosis. Tumour growth is not just a result of uncontrolled cell proliferation but also of reduced apoptosis. High levels of interleukin-6 (IL-6) are associated with metastatic BC and correlated with poor survival as it promotes growth of tumour-initiating cells during early tumorigenesis protecting these cells from apoptosis. Therefore, this study aims at investigating the potential of anti-IL-6 monoclonal antibodies to suppress IL-6 proliferative/anti-apoptotic activities in intact tumour microenvironment of BC. Fresh sterile tumour and normal breast tissue specimens were taken from 50 female Egyptian patients with BC undergoing radical mastectomy. A unique tissue culture system designed to provide cells of each intact tumour/normal tissue sample with its proper microenvironment either supplemented or not with anti-IL-6 monoclonal antibodies. To evaluate the apoptotic activity of anti-IL-6 as a novel candidate for BC treatment strategy, we compared its effects with those obtained using tumour necrosis-related apoptosis-inducing ligand TRAIL as an established apoptotic agent. Our results revealed that levels of either anti-IL-6- or TRAIL-induced apoptosis in the tumour or normal tissue cultures were significantly higher than those in their corresponding untreated ones (P < 0.001). No statistically significant differences have been found between apoptosis levels induced by anti-IL-6 monoclonal antibodies and those induced by TRAIL. Recombinant anti-IL-6 monoclonal antibodies could represent a novel effective element of immunotherapeutic treatment strategy for BC. The selectivity and anti-apoptotic potential of anti-IL-6 is highly hopeful in IL-6- abundant BC tumour microenvironment.

Stem Cell Factor and Interleukin-31 Expression: Association with IgE among Egyptian Patients with Atopic and Nonatopic Bronchial Asthma.

Bronchial asthma is a chronic inflammatory disorder which remains a significant cause of morbidity. Recently, it has been reported that the stem cell factor (SCF) and interleukin-31 (IL-31) may play a major role in bronchial asthma. The aim of the current study was to study the association of the stem cell factor and interleukin-31 expression with serum immunoglobulin E among Egyptian patients with atopic and nonatopic bronchial asthma. After measuring serum IgE using total enzyme linked immunosorbent assay (ELISA), Ribonucleic acid (RNA) was isolated to determine gene expression of SCF and IL-31 by real-time polymerase chain reaction (PCR). The levels of SCF mRNAs in atopic asthmatic patients' PBMCs were significantly higher than those in controls (p = 0.0001**) and nonatopic asthmatics (p = 0.0001**). There was a high statistical significant difference also with regard to IL-31 between atopic asthmatics and controls (p = 0.0001**) and between them and nonatopic patients (p = 0.014*). There was a strong significant direct correlation between SCF, IL-31 (r = 0.827 and p = 0.0001**) and between both of them and IgE in asthmatics (r = 0.543 and p = 0.0001**) (r = 0.443 and p = 0.0001**), respectively. A direct correlation between SCF, IL-31 and FEV-1/ FVC %, CRP and wheezing existed. These findings suggest that both SCF and IL-31 play an important role in mediating inflammation and enhancing severity of atopic asthma. Augmented inhaled glucocorticoid therapy was associated with significant reductions in SCF and IL-31 mRNA expression as well as improvements in lung function, symptom scores and bronchial hyperresponsiveness to methacholine (PD20) in atopic and nonatopic asthmatics.

Th17/Treg imbalance in opioids and cannabinoids addiction: relationship to NF-κB activation in CD4+ T cells.

Opioids are widely used for the treatment of severe pain. However, opioids, particularly morphine, is known to cause immunosuppression. This study investigated the impact of morphine and cannabinoids addiction on CD4+ T cell mediated immunity. We hypothesize that, accompanied immunosuppression is attributed to reduced T cell activation with an extent of affection to the cytoplasmic activity of the biologically active transcription factor nuclear factor-κB (NF-κB) which play crucial role in T-cell activation. A disturbance in cytokine balance, in particular, interleukin-17 (IL-17)/interleukin-10 (IL-10) production may also act as a mechanism of immunosuppression. Peripheral blood CD4+ T cells from 45 chronic morphine and cannabinoid addicts and 10 controls with no current or past history of drug abuse; were stimulated by anti-CD3 antibody plus phytoheamagglutinin (PHA). Activation of the NF-κB signaling pathway was examined by analyzing NF-κBp65 in a solid phase sandwich ELISA. IL-17/IL-10 balance was assessed using quantitative ELISA on cultured CD4+ T cells supernatants. We found that, morphine and cannabinoids inhibited NF-κB signaling in activated T cells of addicts, whereas it enhanced activated T cell apoptosis as measured by quantitative in vitro determination of cytoplasmic histone-associated DNA fragmentation following induced cell death. These effects of morphine and cannabinoids T cell suppression were accompanied by elevation of IL-10 level and concomitant reduction in IL-17 secretion from cultured CD4+ T cells. We concluded that Th17/Treg imbalance may be attributed to inhibited NF-κB activity in CD4+ T cells under the effect of morphine and cannabinoids addiction.

Liposome-based HCV vaccine enhances protective cellular immunity and IFN-gamma secretion in mice.

HCV is a worldwide health problem and with the lack of effective treatment, vaccination becomes an urgent task, especially in developing countries. The effective vaccine should elicit long-lasting antibodies but most importantly induce a vigorous, multi-specific cellular immune response that includes both helper and cytotoxic T lymphocytes. Advances in liposome technology account for much of the progress in vaccine delivery systems. Therefore, this study aimed at investigating the potential immunogenicity of HCV core antigen, and assessing the influence of the novel antigen carried on liposomes on T cell proliferation and IFN-gamma production as potent markers of cellular immune response. Several formulations for immunization were prepared, including liposomal encapsulation of the Ag. The study was conducted on a total of 95 female inbred (C57B1/6J) mice divided into five groups including a control group. Spleen lymphocytes were evaluated for cellular proliferation using 3-(4, 5-Dimethylthiazol-2-YI)-2, 5-Diphenyltetrazolium Bromide (MTT) assay and for secretion of IFN-gamma by ELISA. Mice injected with liposomes carrying HCV core Ag (group 1) showed a highly significant increase in splenocytes proliferation (spontaneously and after stimulation with the Ag) compared to all other groups, with a stimulation index (S.I) of 1.47 (P < 0.001). The second highest cellular proliferation was noticed in mice injected with core Ag and CFA (group 2) (S.I = 1.29) with a significant difference from group I (P = 0.001). Mice injected with core Ag alone showed a non-significant difference from the control group (P = 0.126). IFN-gamma level was the highest in liposomal Ag group with a highly significant difference; both spontaneously (56.3 pg/L) and with stimulation (68.32) (P < 0.001) followed by mice injected with core Ag with CFA. In conclusion, Liposomal formulation of HCV peptide vaccine is effective as direct in vivo antigen loading and activation of T cells leading to protective HCV antiviral responses.