ß-glucan nanoparticles alleviate acute asthma by suppressing ferroptosis and DNA damage in mice.

Asthma is a severe respiratory disease marked by airway inflammation, remodeling, and oxidative stress. ß-Glucan (BG), a polysaccharide constituent of fungal cellular structures, exhibits potent immunomodulatory activities. The investigational focus was on the anti-asthmatic and anti-ferroptotic properties of beta-glucan nanoparticles (BG-NPs) in a murine model of allergic asthma induced by ovalbumin (OVA). BG was extracted from Chaga mushrooms (Inonotus obliquus), and its BG-NPs were characterized utilizing techniques including FT-IR, UV visible spectroscopy, zeta potential analysis, DLS, XRD, and TEM. The Balb/C mice were allocated into five groups: control, untreated asthmatic, dexamethasone (Dexa)-treated (1 mg/kg), BG-treated (100 mg/kg), BG-NPs-treated (45 mg/kg), and BG-treated (100 mg/kg). Treatment with BG-NPs markedly diminished the entry of inflammatory cells into the respiratory passage, serum IgE concentrations, DNA damage, and markers of oxidative stress through the reduction of malonaldehyde (MDA) levels and enhancing the levels of reduced glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT). Furthermore, BG-NPs reduced iron deposition and promoted the transcriptional activity of the GPx4 gene in pulmonary cells, attenuating ferroptosis. The results demonstrated that BG-NPs reduced asthma by inhibiting oxidative stress, inflammation, DNA damage, and ferroptosis. Our results suggest that BG-NPs could be used as potential treatments for allergic asthma.

4-Hydroxychalcone attenuates ovalbumin-induced allergic airway inflammation and oxidative stress by activating Nrf2/GPx4 pathway.

Asthma is a lung condition characterized by impaired respiratory function and an apparent infiltration of inflammatory cells. Chalcones are substances that have attracted considerable interest in the disciplines of pharmaceutical chemistry and drug discovery due to their diverse biochemical processes, such as antioxidant, anti-inflammatory, anticancer, antibacterial, and others, but whether they can be used in asthma treatment has yet to be investigated. This study aimed to investigate the immunomodulatory effect of 4 hydroxychalcone (4-HC) against allergic asthma in mice. In this research, we investigated how 4-HC affected asthmatic behavior, leukocyte infiltration, histopathological alterations, oxidative stress, immunoglobulin E (IgE) production, and airway inflammation. Moreover, ELISA and immunohistochemistry (IHC) were used to measure the expression of Nrf2 and GPx4. 4-HC treatment significantly decreased lung oxidative stress, inflammatory cell infiltration, and IgE levels. According to our findings, we imply that 4-HC may be utilized as an anti-asthmatic agent through the upregulation of Nrf2/GPx4 signaling pathway.

Immune response, oxidative stress, and histological changes of Wistar rats after being administered with Parascaris equorum antigen.

Worldwide, particularly in developing nations, helminth infections are the leading causes of livestock illness and mortality. Parascaris (P.) equorum, a parasitic worm from the Ascarididae family, significantly impacts the production, health, and working performance of equines. This study aimed to investigate the impact of intraperitoneal sensitization of P. equorum on the immune system, oxidative stress, and histology in Wistar rats. After acclimatization for 7 days, we divided the rats into five groups, each consisting of six rats. Group I, serving as the control, was administered distilled water, followed by groups II (day 7), III (day 14), IV (day 21), and V (day 33). The rats were euthanized every day mentioned (Days 7-33). On day 0, a dosage of 1ml/100 gm rat (containing 500 µg/ml protein content) emulsified crude antigen extract with an incomplete Freund's adjuvant (1:1 volume), followed by a second dose of the same antigen concentration on day 7. To assess the allergenicity of this nematode, we measured a whole blood profile, serum levels of IFN-γ, IL-5, IL-10, IL-13, and IL-33, total immunoglobulins IgE and IgG, and oxidative stress markers. Also, we examined histological changes in the liver, kidney, and spleen. The results showed that values of total leukocyte count, granulocytes, monocytes, and lymphocytes were significantly (P < 0.05) increased on day 14 post-infection relative to other days of investigation. It was found that the levels of total immunoglobulins (IgE and IgG) and cytokines (INF-γ, IL-5, IL-13, and IL-33) on days 14 and 21 were significantly higher than in the control group. At all periods of the experiment, the injected group exhibited significantly higher concentrations of MDA and NO compared to the control group (P < 0.05). Conversely, GSH and CAT levels (P < 0.05) dropped significantly on days 7, 14, and 21. Different rat tissues showed alterations. Ultimately, this study described the detrimental effects of P. equorum crude antigen administration on the immune system, oxidative states, and histological changes of Wistar rats at various intervals.

Impact of sub chronic administration of deltamethrin on autoimmune activity in rat.

Deltamethrin (DLM) is a newer kind of insecticide that is used on pets, livestock, and crops, as well as to combat malaria vectors and household pests. It belongs to the synthetic pyrethroid group and is being promoted as an alternative to organophosphate chemicals due to its persistent and destructive effects. The current study aimed to evaluate the impact of sub-chronic oral exposure to DLM on autoimmune activity in rats. Three groups of male albino rats (15 rats/group) including the control group, the ethanol-treated group (1 ml/rat), and the DLM-treated group (5 mg/kg b.w). Samples of blood were taken from all groups at 4-, 8- and 12-week intervals for the determination of hematological, cytokines, and immunological parameters. T lymphocyte subsets and Treg lymphocytes were determined in serum using flow cytometric acquisition. The results revealed that DLM significantly increased TNF-α, IL-33, IL-6, IL-17, IgG, IgM, WBCs, differential count, and platelets while decreasing Hb concentration and RBCs. Additionally, DLM decreased the number of T-cell subsets (CD3, CD4, CD5, and CD8) and Treg lymphocytes. All of these impacts became more severe over time. It is possible to conclude that the sub-chronic oral exposure to DLM disturbed autoimmune activity through the disturbances in immunological indices, CDs subset Treg lymphocytes.

Photodynamic therapy mediated by methylene blue-loaded PEG accelerates skin mouse wound healing: an immune response.

PURPOSE: Conventional approaches for enhancing wound healing may not always yield satisfactory results. Instead, we test the effectiveness of a newly developed photodynamic therapy (PDT) that uses methylene blue (MB) loaded with polyethylene glycol (PEG) (MB-PEG) hydrogel to accelerate wound healing process in mice. METHODS: A dorsal skin incision with 6 mm punch which topically subjected to MB-PEG hydrogel and a low-level laser light of red light to assess the regeneration process of wounded skin. A total of 63 adult male CD1 mice divided into normal group (no treatment) and other wound groups received different treatments of laser (650 ± 5 nm and power intensity of 180 mW/cm2), MB-PEG, or PDT (MB-PEG followed by laser). The wound healing parameters were investigated by histological examination of the skin and measuring of proinflammatory cytokines at the early stage (48 h) and a late one on day 21. RESULTS: at 48 h, the score of tissue granulation, inflammation, and angiogenesis process were markedly improved in wounded groups that received MB + PEG combined with laser compared to the group treated with laser alone. On day 21, a significant improvement of the inflammation was detected in the group treated with MB + PEG plus laser compared to the other groups. At 48 h, the upregulated serum levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1ß in the wound group were significantly (P < 0.001) reduced in the group treated with MB + PEG combined with laser. CONCLUSION: MB-PEG based hydrogel improves and accelerates wound closure in the context of laser compared to either single treatment.

ß-glucan mitigates ovalbumin-induced airway inflammation by preventing oxidative stress and CD8+ T cell infiltration.

BACKGROUND: Bronchial asthma is a severe respiratory condition characterized by airway inflammation, remodeling, and oxidative stress. ß-Glucan (BG) is a polysaccharide found in fungal cell walls with powerful immunomodulatory properties. This study examined and clarified the mechanisms behind BG's ameliorativeactivitiesin an allergic asthma animal model. METHOD: BG was extracted from Chaga mushroom and characterized using FT-IR, UV-visible, zeta potential, and 1H NMR analysis. The mice were divided into five groups, including control, untreated asthmatic, dexamethasone (Dexa)-treated (1 mg/kg), and BG (30 and 100 mg/kg)-treated groups. RESULTS: BG treatment reduced nasal scratching behavior, airway-infiltrating inflammatory cells, and serum levels of IgE significantly. Additionally, BG attenuated oxidative stress biomarkers by lowering malonaldehyde (MDA) concentrations and increasing the levels of reduced glutathione (GSH), glutathione peroxidase (GPx), and catalase (CAT). Immunohistochemical and flow cytometric analyses have confirmed the suppressive effect of BG on the percentage of airway-infiltrating cytotoxic CD8+ T cells. CONCLUSION: The findings revealed the role of CD8+ T cells in the pathogenesis of asthma and the role of BG as a potential therapeutic agent for asthma management through the suppression of airway inflammation and oxidative stress.

Echinochrome exhibits anti-asthmatic activity through the suppression of airway inflammation, oxidative stress, and histopathological alterations in ovalbumin-induced asthma in BALB/c mice.

Asthma is a chronic pulmonary disease with marked infiltrating inflammatory cells and reduced respiratory performance. Echinochrome (Ech) is a dark-red pigment isolated from the sea urchin spines, shells, and ova. It has antioxidant, antimicrobial, and anti-inflammatory properties, but whether it can be used in asthma treatment has yet to be investigated. In this research, we aimed to study the inhibitory actions of Ech on allergic asthma symptoms in mice. Mice were divided into 4 groups (n = 8 for each): control, ovalbumin-challenged, and Ech-treated (0.1 and 1 mg/kg). At the end of the experiment, nasal scratching, lung oxidative stress, airway inflammation, and remodeling were assessed. In ovalbumin-challenged BALB/C mice, treatment with Ech significantly decreased nasal scratching, lung oxidative stress, inflammatory cell infiltration, mucus hyperproduction and hyperplasia of goblet cells, IgE levels, and inflammatory cytokines. It also inhibited NF-κB phosphorylation. This is the first study to investigate the immunomodulatory effect of Ech against allergic asthma in mice. According to our findings, we imply that Ech may be utilized as a treatment for allergic asthma.

Echinochrome Ameliorates Physiological, Immunological, and Histopathological Alterations Induced by Ovalbumin in Asthmatic Mice by Modulating the Keap1/Nrf2 Signaling Pathway.

Asthma is a persistent inflammatory disease of the bronchi characterized by oxidative stress, airway remodeling, and inflammation. Echinochrome (Ech) is a dark-red pigment with antioxidant and anti-inflammatory activities. In this research, we aimed to investigate the effects of Ech against asthma-induced inflammation, oxidative stress, and histopathological alterations in the spleen, liver, and kidney in mice. Mice were divided into four groups (n = 8 for each): control, asthmatic, and asthmatic mice treated intraperitoneally with 0.1 and 1 mg/kg of Ech. In vitro, findings confirmed the antioxidant and anti-inflammatory activities of Ech. Ech showed antiasthmatic effects by lowering the serum levels of immunoglobulin E (IgE), interleukin 4 (IL-4), and interleukin 1ß (IL-1ß). It attenuated oxidative stress by lowering malondialdehyde (MDA) and nitric oxide (NO) contents and increasing reduced glutathione (GSH), superoxide dismutase (SOD), glutathione-s-transferase (GST), and catalase (CAT) in the liver, spleen, and kidney. Moreover, it protected asthma-induced kidney and liver functions by increasing total protein and albumin and decreasing aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, urea, and uric acid levels. Additionally, it ameliorated histopathological abnormalities in the lung, liver, spleen, and kidney. Additionally, molecular docking studies were used to examine the interactions between Ech and Kelch-like ECH-associated protein 1 (Keap1). PCR and Western blot analyses confirmed the association of Ech with Keap1 and, consequently, the regulatory role of Ech in the Keap1-(nuclear factor erythroid 2-related factor 2) Nrf2 signaling pathway in the liver, spleen, and kidney. According to our findings, Ech prevented asthma and its complications in the spleen, liver, and kidney. Inhibition of inflammation and oxidative stress are two of echinochrome's therapeutic actions in managing asthma by modulating the Keap1/Nrf2 signaling pathway.

A new perspective on quantitative assessment of photodynamic therapy mediated hydrogel nanocomposite in wound healing using objective biospeckle and morphological local-gradient.

Skin wounding is a serious public health issue, especially when considering factors that accelerate tissue recovery. Consequently, the use of photodynamic therapy (PDT) as an effective wound-healing treatment has attracted more scientific attention. Although assessing the wound healing rate is crucial for appropriate monitoring of the probability of wound healing and evaluating the treatment efficiency, the currently used techniques lack the ability to provide such information. Therefore, this study has two aims, first, it contributes to the development of a new image-guided biospeckle system for quantitative monitoring of skin wound healing rate. Second, it evaluates the potential of using a novel synthesized PDT-mediated polyethylene glycol fabric with methylene blue (PEG-MB) hydrogel nanocomposite in accelerating wound healing. The proposed imaging system initially acquires raw biospeckle images from the wound regions of adult healthy albino mice treated with the synthesized hydrogel nanocomposite. Each raw biospeckle image is then converted into maps of morphological local-gradient matrices implemented from the combination of dilation and erosion operations at different radii up to 25 pixels. Subsequently, their intensity histogram statistics are computed, taking central moments as the feature set. Final characterization is achieved via a linear combination of the biospeckle statistics maintaining as much variance as possible using principal component analysis (PCA). The results confirmed by cytokine concentration measurement and histological investigation demonstrate that the innovative biospeckle image-guided system is ideal for investigating wound healing and suggest the potential of the hydrogel nanocomposite as an active dressing.

MicroRNA-26a systemic administration attenuates tumor formation in hepatocellular carcinoma mouse model.

MicroRNA (miRNA)-26a is one of the tumor suppressor genes that has been down regulated during the development of hepatocellular carcinoma (HCC). This work was conducted to evaluate the possible preventive effect of exogenous miRNA-26a administration on diethylnitrosamine (DEN)-mediated HCC. Balb/C mice were intraperitoneally injected with saline (Normal group), DEN (HCC group) or miRNA-26a (HCC+miRNA-26a group). On week 8, 12, 16 and 20, the concentrations of alpha-fetoprotein (AFP), des-gamma carboxyprothrombin (DCP), the levels of helper T cells-associated cytokines, and the vascular endothelial growth factor (VEGF), were measured. Flow cytometry determined the frequencies of regulatory T (Treg) cells. The concentrations of AFP, DCP and VEGF, as well as the frequency of Treg cells showed significantly lower values following miRNA-26a administration than in HCC group. miRNA-26a administration has reduced the levels of IL (interleukin)-2 and TNF (tumor necrosis factor)-α, in contrast, IL-10 level was markedly elevated in comparison to HCC model at all experimental time points. The restore of miRNA-26a function significantly (P<0.001) down regulated the expression levels of survivin & caspase-3 compared to HCC group. The obtained data introduce an evidence for the suppressive impact of miRNA-26a on liver tumor formation and its possible manipulation as a therapeutic design for HCC.

Culture of Care Enhancement in Egypt: The Impact of Laboratory Animal Science Training on Participants' Attitudes.

Cairo University was the first academic institution in Egypt to establish an Institutional Animal Care and Use Committee (IACUC), as mandated by the World Organisation for Animal Health (OIE). Animal-based research should be performed in accordance with international regulations to monitor the humane care and use of the laboratory animals. Until 2018, the formal training of researchers in the appropriate and correct methods of animal handling during sampling and administration, as well as their husbandry demands, was an uncommon practice in Egypt. In 2018, the Egyptian Association for Animal Research Advancement (EAARA) organised the first international course in laboratory animal science (LAS), in collaboration with Utrecht University (The Netherlands) and the Faculty of Science, Cairo University, to raise researchers' awareness and increase their knowledge of the principles that govern the humane use and care of laboratory animals. A total of 26 researchers from a number of fields (veterinary medicine, dentistry, science, medicine, pharmacy and agriculture) enrolled in the course. In the responses to the post-course questionnaire, 24 (92.3%) participants stated that the principles of animal welfare (Three Rs) were well explained. In addition, 18 (69%) participants found that the course improved their skills in animal sampling and handling. Of the 26 participants, 22 (84.6%) became aware of their responsibility towards their experimental animals and agreed that the different methods of euthanasia were well explained. In conclusion, the general assessment of the course revealed a positive outcome regarding the culture of animal care; the course was repeated a year later, and several participants were enlisted as trainers in this second course.

Microsatellite instability profiling in Egyptian bladder cancer patients: A pilot study.

Microsatellite alterations have been implicated in the pathogenesis of many cancers; however, they are still not well addressed in the bladder cancer (BC) of Egyptian population. We assessed microsatellite instability (MSI) profile and loss of heterozygosity (LOH) using 13 microsatellite markers in tumor tissue samples and urine sediments obtained from 30 Egyptian patients with BC. The concordance between MSI in tumor tissue and urine samples was determined, and correlated to relevant clinicopathologic features. We found that MSI was more frequent than LOH (100% and 46.7%, respectively). D16S310, MBP, and IFN-α showed the highest MSI frequency in urine samples (70%, 70%, and 66.67%, respectively), while MBP, ACTBP2, and D9S171 (66.67%, 63.33%, and 60%, respectively) were the most frequently detected in tumor tissues. All assessed MSI markers correlated significantly with pathologic subtype (being more frequent in TCC) and with hematuria. The concordance between tissue and urine samples was statistically significant for D16S476, D9S171, FGA, and ACTBP2 (P = 0.04, 0.015, 0.02, and 0.007, respectively). When we combined D16S476 and D9S171, the sensitivity, specificity, positive predictive value, and negative predictive value for the diagnosis of BC were 80.0%, 75.0%, 82.8%, and 71.4%, respectively. Accordingly, we concluded that MSI in urine sediments could be a potential tool for the diagnosis of BC.

Development of new lateral-flow immunochromatographic strip using colloidal gold and mesoporous silica nanoparticles for rapid diagnosis of active schistosomiasis

Objective: To develop a new sandwich based lateral flow immunochromatographic strip for rapid detection of circulating Schistosoma mansoni antigen in serum and urine samples of patients with active schistosomiasis. Methods: This lateral flow immunochromatographic strip was prepared by using anti-Schistosoma mansoni soluble egg antigen monoclonal antibody conjugated gold nanoparticles (MAb-AuNPs) as antigen-detecting antibody, while crystalline material (MCM)-41-MAb bioconjugate was immobilized at the test line as antigen-capturing antibody. Both antigen capturing and detecting antibodies formed sandwich complexes with circulating Schistosoma mansoni antigen in the positive samples. Sandwich complexes immobilized at the test line gave distinct red color. The assay reliability was examined by using urine and serum samples of 60 Schistosoma mansoni infected patients, 20 patients infected with parasites other than Schistosoma, and 20 healthy individuals as negative controls. Results were compared with those obtained via sandwich enzyme linked immunosorbent assay (ELISA). Results: The detection limit of circulating Schistosoma mansoni antigen by lateral flow immunochromatographic strip was lower (3 ng/mL) than the detection limit by ELISA (30 ng/mL). The sensitivity and specificity of lateral flow immunochromatographic strip in urine samples were 98.3％ and 97.5％, respectively compared to 93.5％ and 90.0％ by ELISA. In serum samples, they were 100.0％ and 97.5％, respectively compared to 97.0％ and 95.0％ by ELISA. The strip test took approximately 10 min to complete. Conclusions: This new lateral flow immunochromatographic strip offers a sensitive, rapid, and field applicable technique for diagnosis of active schistosomiasis.

Role of relevant immune-modulators and cytokines in hepatocellular carcinoma and premalignant hepatic lesions.

AIM: To assess the levels of different immune modulators in patients with hepatocellular carcinoma (HCC), in relation to other hepatic diseases. METHODS: Eighty-eight patients were included in the current study and represented patients with HCC (20), liver cirrhosis (28) and chronic hepatitis (CH; 25), and normal controls (NC; 15). Peripheral blood was isolated for immunophenotyping of active myeloid dendritic cells (mDCs; CD1c and CD40), mature inactive myeloid cells (CD1c and HLA), active plasmacytoid cells (pDCs; CD303 and CD40), mature inactive pDCs (CD30 and HLA), active natural killer (NK) cells (CD56 and CD161), active NK cells (CD56 and CD314) and inactive NK cells (CD56 and CD158) was done by flow cytometry. Serum levels of interleukin (IL)-2, IL-10, IL-12, IL-1ß, interferon (IFN)-α, IFN-γ and tumor necrosis factor (TNF)-αR2 were assessed by ELISA. RESULTS: Active mDCs (CD1C+/CD40+) and inactive mDCs (CD1c+/HLA+) were significantly decreased in HCC patients in relation to NC (P < 0.001). CD40+ expression on active pDCs was decreased in HCC patients (P < 0.001), and its level was not significantly changed among other groups. Inactive pDCs (CD303+/HLA+), inactive NKs (CD56+/CD158+) and active NKs (CD56+/CD161+) were not statistically changed among the four groups studied; however, the latter was increased in CH (P < 0.05). NKG2D was statistically decreased in HCC, CH and cirrhosis (P < 0.001), and it was not expressed in 63% (12/20) of HCC patients. There was significant decrease of IL-2, IFN-α and IFN-γ (P < 0.001), and a significant increase in IL-10, IL-1ß, and TNF-αR2 (P <0.01, P < 0.001 and P < 0.001; respectively) in HCC patients. There was inverted correlation between IL-12 and IL-1ß in HCC (r = -0.565, P < 0.01), with a strong correlation between pDCs (CD303+/CD40+) and NKs (CD56+/CD161+; r = 0.512, P < 0.05) as well as inactive mDCs (CD1c+/HLA+) and inactive NK cells (CD56+/CD158+; r = 0.945, P < 0.001). CONCLUSION: NKG2D, CD40, IL-2 and IL-10 are important modulators in the development and progression of HCC.

Therapeutic efficacy of osthole against dinitrobenzene sulphonic acid induced-colitis in rats.

Several mediators were associated with the pathogenesis of inflammatory bowel disease such as oxidative stress through the production of reactive oxygen metabolites, neutrophils infiltration and release of pro-inflammatory cytokines. This study was designed to investigate the therapeutic efficacy of osthole against dinitrobenzene sulfonic acid (DNBS) induced-colitis in rats through its anti-oxidant and anti-inflammatory properties. Colitis was induced in rats by single intracolonic instillation of (250 µl DNBS-25 mg/rat). Then 4 days later, rats were received oral administration of either (osthole 50 mg/kg), (sulfasalazine 500 mg/kg) or both in combination for 7 consecutive days. Body weight, some hematological parameters, colonic malondialdehyde (MDA) and myeloperoxidase activity (MPO), antioxidant parameters, colon injury and mucosa architectures were assessed. T helper (Th1)-related cytokines [Tumor necrosis factor alpha (TNF-α) and interferon-gamma (INF-γ)], Th2-relarted cytokines (interleukin-4 [IL-4 and IL-10], and Th-17 related cytokines [IL-17] were determined by ELISA. Osthole significantly improved the loss in body weight. That was accompanied with a remarkable amelioration of the disruption of the colonic architecture as well as a significant improvement in the antioxidant defense system. A reduction in MPO and MDA was observed in flamed colon. Treatment with either osthole or combination therapy showed suppressive activities on pro-inflammatory Th2-related cytokines and upregulation of anti-inflammatory Th2-related cytokines The results of this study suggest that osthole exert beneficial therapeutic effect in experimental colitis and improved the efficacy of the synthetized drugs such as sulfasalazine. Therefore, osthole may have a valuable sound in the treatment of inflammatory bowel disease.

Pathogenic Potential of Fresh, Frozen, and Thermally Treated Anisakis spp. Type II (L3) (Nematoda: Anisakidae) after Oral Inoculation into Wistar Rats: A Histopathological Study.

The third-stage (L3) larvae of Anisakis are the etiological agents of human anisakiasis caused by consumption of raw or undercooked seafood infected with anisakid nematodes. Infection with these worms is associated with abdominal pain, nausea, and diarrhea and can lead to massive infiltration of eosinophils and the formation of granulomas in the gastrointestinal tract if the larvae are not removed. Food allergy affects populations worldwide, and despite several reports on the presence of the potentially zoonotic nematodes among edible fishes in Egypt, there are few immunological and molecular studies investigating the epidemiology of these parasites. Anisakidosis, a human infection with nematodes of the family Anisakidae, is caused most commonly by Anisakis spp. In the present study, seventy specimens of the European seabass Dicentrarchus labrax commercialized in Alexandria city along the Mediterranean Sea were acquired during the period from July to December, 2015. Fish were necropsied and dissected to investigate the presence of nematode larvae. Thirty fish (42.9%) of the total were parasitized by nematode larvae which were morphologically identified as Anisakis spp. Type II (L3) according to light and scanning electron microscopy. The pathogenic potential of oral inoculation of fresh, frozen, and thermally treated larvae into Wistar rats was elucidated by histological examination of their thymus and spleen. Results obtained indicated that neither cooling nor freezing of the parasite could destroy their allergenic capacity. So, it is important to create a wider awareness of this potential risk to human health. It is becoming increasingly likely that the impact of Anisakis spp. on human health has been underestimated, and it is perhaps time to consider more sweeping measures than those currently enforced to protect the public health.

Interleukin-6 induces secretion of tissue inhibitors of etalloproteinases by breast carcinoma cells.

Interleukin (IL)-6 can induce matrix metalloproteinases (MMPs) expression, which may be critical factors involved in tumor metastasis. Tissue inhibitors of metalloproteinases (TIMPs) are important inhibitory enzymes of MMP. This study was designed to investigate the effect of recombinant IL-6 on the MMP/TIMP expression in MDA-MB-231 breast cancer cell line in a dose dependent manner (10, 25 and 50ng/ml) in comparison to non-treated breast cancer cells (control). The data demonstrated that low dose (10 ng/ml) of IL-6 failed to induce TIMP-1 and -2 production by breast cancer cells compared to control cells whereas moderate (25 ng/ml) and high (50ng/ml) exposure levels promoted a significant expression of TIMP-1 (P<0.01 and P<0.0001) respectively as compared to control cells. TIMP-2 was significantly released (P<0.0001) from breast cancer cells higher than in control cells at moderate and high exposure levels of IL-6. This up-regulation of TIMP-1 and -2 was accompanied with undetectable levels of MMP-1, -2, -3, -8, -9, -10, and -13. Furthermore, IL-6 potentially increased the invasion potency of cancer cells significantly (P<0.05 and P<0.01) at moderate and high exposure levels respectively. These findings suggest that IL-6 could promote the invasion potency of breast cancer cells by inducing secretion of TIMP-1 and -2, causing a disturbance in TIMP/MMP balance.

Cytokine signature and antibody-mediated response against fresh and attenuated Anisakis simplex (L3) administration into Wistar rats: implication for anti-allergic reaction.

The third larval stage (L3) of Anisakis simplex (Anisakidae) is one of the zoonotic parasitic nematodes in the musculature and visceral organs of marine fishes belonging to family Moronidae. The consumption of these high-commercial-value fish is widespread in many countries around the Mediterranean Sea including Egypt. The presence of these larvae in fish muscles poses a potential consumer hazard due to the parasite's ability to cause anisakidosis. Forty-two out of 60 (70%) of the European seabass Dicentrarchus labrax were found to be naturally infected by L3 of A. simplex in the form of encapsulated juveniles in the fish musculature. Morphological examination of recovered parasites by light and scanning electron microscopy showed that, in general, all specimens examined closely resembled A. simplex (L3). To evaluate the allergenicity of this nematode, white blood cell count; levels of T helper 1 (Th1) [interferon (IFN)-γ and tumor necrosis factor (TNF)-α)], Th2 [IL-4, IL-5, and IL-6], and Th17 [IL-17] related cytokines; total IgE and IgG antibodies; and nitric oxide (NO) were measured in the plasma of Wistar rats sensitized by oral inoculation with fresh, frozen, and heat-treated A. simplex L3 or rats intraperitoneally injected with L3 crude extract. Rats sensitized with fresh and frozen L3 larvae produced significantly higher levels of IFN-γ, IL-5, IL-17, and total IgE as compared to control rats. Heat-treated larvae administration resulted in a significant rise of IFN-γ, TNF-α, IL-5, and total IgE in comparison to control rats. Intraperitoneal sensitizations enhanced release of IFN-γ, TNF-α, and total IgE. Oral sensitization led to a significant production of NO. Thereby, frozen or cooked larval L3 cannot inhibit the release of Th-related cytokines and IgE, which might impact on the overall anti-parasitic immunity.

Immunodiagnosis of Egyptian human fasciolosis gigantica using Fas1 and Fas2 cysteine proteinase antigens.

Fasciolosis caused by Fasciola gigantica is one of the major public health problems in the world including Egypt. Immunodiagnostic methods are more applicable for their better sensitivity and specificity than other methods. The present study was conducted to cysteine proteinase (CP) antigens of F. gigantica in IgG-ELISA to diagnose human fasciolosis. IgG-ELISA with 2 cysteine proteinases of 27 kDa (Fas1) and 29 kDa (Fas2), obtained from the regurgitated materials of adult worms, were evaluated using serum samples from 90 Egyptian patients infected with F. gigantica, 55 patients with other parasitic infections and 50 healthy volunteers. The diagnostic sensitivity and specificity of Fas1 for detection of F. gigantica infection were 91.1% and 89.1%, respectively. The positivity of the assay was 95%. The positive and negative predicted values were 91% and 86%, respectively. These data suggest that IgG-ELISA with Fas1 is highly sensitive and specific assay and could be used for the immunodiagnosis of human fasciolosis.