Evaluation of the efficacy of gold nanoparticles on Giardia lamblia infection in experimental animals.

Giardia lamblia (G. lamblia) is an important cause of severe malabsorption, weight loss, physical and mental retardation especially in infants and children throughout the world. Metronidazole (MTZ) is the standard drug used for their treatment which possesses several drawbacks with low efficacy. Gold nanoparticles possess a broad-spectrum antimicrobial activity and could be considered as a future alternative to many microbial agents. This study aimed to evaluate the anti-Giardia effect of gold nanoparticles as an alternative to MTZ. This study was done on 70 experimentally albino rats that were divided into three main groups with seven subgroups (each of 10 rats). The effect of MTZ and gold nanoparticles as single or combined therapy were evaluated. The effect was assessed by counting Giardia fecal cysts in the stool and trophozoites in the intestinal wash, histopathological, transmission and scanning electron microscopic examinations of the small intestinal tissues. Toxic tests of biochemical parameters of liver and kidney function were also performed. A significant reduction of the parasite number in the stool and small intestinal sections was apparent in treated infected rats compared with the infected non-treated ones. Gold nanoparticles showed the best result and the highest effect in the eradication of the parasite from the stool and the intestine with marked improvement in the intestinal mucosal injury caused by G. lamblia trophozoites. Gold nanoparticles had a toxic effect on the liver, with no kidney toxicity. Nanogold can be considered as a potential therapeutic agent and as a promising alternative therapy for G. lamblia infection. Further studies using various dosages with different durations of treatment with gold nanoparticles can be tested on Giardia lamblia infection.

Dual targeting nanoparticles based on hyaluronic and folic acids as a promising delivery system of the encapsulated 4-Methylumbelliferone (4-MU) against invasiveness of lung cancer in vivo and in vitro.

Lung cancer is the most common cause of cancer death worldwide. Thereby, new treatment strategies as targeting nano-therapy present promising possibilities to control the aggressiveness of lung cancer. Dual CD44 and folate receptors targetable nanocapsule based on folic-polyethylene glycol-hyaluronic (FA-PEG-HA) were fabricated to improve the therapeutic activity of 4-Methylumbelliferone (4-MU) toward lung cancer. In this study, we fabricate 4-MU Nps as a hybrid polymeric (protamine) protein (albumin) nanocapsule, then functionalized by targeting layer to form 4-MU@FA-PEG-HA Nps with encapsulation efficacy 96.15%. The in vitro study of free 4-MU, 4-MU Nps and 4-MU@FA-PEG-HA Nps on A549 lung cancer cells reveal that the 4-MU Nps and 4-MU@FA-PEG-HA Nps were more cytotoxic than free 4-MU on A549 cells. The observed therapeutic activity of 4-MU@FA-PEG-HA Nps on urethane-induced lung cancer model, potentiality revealed a tumor growth inhibition via apoptotic mechanisms and angiogenesis inhibition. The results were supported by Enzyme-linked immunosorbent assay (ELIZA) of transforming growth factors (TGFß1) and serum HA, histopathological analysis as well as immunohistochemical Ki67, CD44, Bcl-2 and caspace-3 staining. Moreover, 4-MU@FA-PEG-HA Nps exhibited a promising safety profile. Hence, it is expected that our developed novel nano-system can be used for potential application on tumor therapy for lung cancer.

Vaccine hesitancy: Beliefs and barriers associated with COVID-19 vaccination among Egyptian medical students.

BACKGROUND: Vaccine hesitancy poses serious challenges for achieving coverage for population immunity. It is necessary to achieve high COVID-19 vaccination acceptance rates and medical students' coverage as future health care providers. The study aimed to explore the level of COVID-19 vaccine hesitancy and determine the factors and barriers that may affect vaccination decision-making. METHODS: A cross-sectional study was carried out among medical students in Tanta and Kafrelsheikh Universities, Egypt. Data collection was done via an online questionnaire during January 2021 from 2133 students. RESULTS: The majority of the participant students (90.5%) perceived the importance of the COVID-19 vaccine, 46% had vaccination hesitancy, and an equal percentage (6%) either definitely accepted or refused the vaccine. Most of the students had concerns regarding the vaccine's adverse effects (96.8%) and ineffectiveness (93.2%). The most confirmed barriers of COVID-19 vaccination were deficient data regarding the vaccine's adverse effects (potential 74.17% and unknown 56.31%) and insufficient information regarding the vaccine itself (72.76%). CONCLUSION: The government, health authority decision-makers, medical experts, and universities in Egypt need to work together and make efforts to reduce hesitancy and raise awareness about vaccinations, consequently improving the acceptance of COVID-19 vaccines.

Prostatic Carcinogenesis: More Insights.

BACKGROUND: Prostatic carcinoma ranks as the second most common malignant tumor and the fifth cause of cancer-related deaths in men. Many studies now focus on the different molecules involved in prostatic carcinogenesis. Maspin and prohibitin (PHB) are suggested to play crucial roles in the development and progression of many cancers; however, their roles in prostatic carcinogenesis have not been fully elucidated. AIM: This work was designed to study the immunohistochemical expression of maspin and PHB in prostatic carcinoma in comparison to their expression in benign prostatic hyperplasia (BPH) to give more insights about their roles in prostatic carcinogenesis. MATERIALS AND METHODS: Archival blocks of 30 cases of prostatic adenocarcinomas and 15 cases of BPH were subjected to histopathological examination and immunohistochemical evaluation of maspin and PHB expression. RESULTS: Maspin showed higher expression in prostatic carcinoma (88.9% of cases) compared to BPH (20% of cases). PHB expression was detected only in prostatic carcinoma (84.4% of cases), while all cases of BPH were negative. The expression of both maspin and PHB showed statistically significant increase with increasing Gleason score (P = 0.0125 and 0.0065 respectively). CONCLUSIONS: Overexpression of maspin and PHB in prostatic carcinoma reflects their vital roles in prostatic carcinogenesis. Their upregulation with increasing Gleason score indicates their prognostic significance. Moreover, PHB may differentiate between prostatic carcinoma and BPH being expressed only by malignant cells.

The role and prognostic value of inducible nitric oxide synthase (iNOS) and interleukin-33 (IL-33) in serous and mucinous epithelial ovarian tumours.

Understanding different mechanisms contributing to the aggressive behaviour of epithelial ovarian cancer (EOC) is a large challenge. Interaction between inflammation, immunity and carcinogenesis occurs in different cancers; however, the potential roles of different molecules involved in these processes in relation to ovarian carcinogenesis were not fully investigated. Inducible nitric oxide synthase (iNOS) and interleukin-33 (IL-33) are implicated in carcinogenesis. iNOS is an NOS isoform that generates nitric oxide, which plays important roles in various stages of carcinogenesis. IL-33 is a cytokine implicated in modulation of anti-tumour immunity and tumour growth. This work aimed at studying the immunohistochemical expression of iNOS and IL-33 in serous and mucinous epithelial ovarian tumours to investigate their role and prognostic significance. Immunohistochemical expressions of iNOS and IL-33 were assessed in 90 patients with epithelial ovarian tumours (45 serous and 45 mucinous tumours, categorized as benign, borderline, and malignant tumours). iNOS and IL-33 showed significantly higher expressions in borderline and malignant serous and mucinous tumours compared to benign ones (p=0.0001). The differences between borderline and malignant tumours were statistically insignificant (p=0.2351&0.6321). iNOS showed significantly higher expression with increasing tumour grade in malignant mucinous tumours (p=0.0011). IL-33 showed significantly higher expression with increasing tumour grade in both malignant serous and mucinous tumours (p=0.0074 and 0.0007). Upregulation of iNOS and IL-33 expression in borderline and malignant epithelial ovarian tumours indicates their involvement in the development and progression of EOC, and their increased expression in less differentiated cancers suggests their association with poor prognosis in this category of tumours.

Atorvastatin and metformin administration modulates experimental Trichinella spiralis infection.

The host-parasite interaction can be altered by the changes in the host environment that may be or may not be in favor of successful invasion by the nematode parasite Trichinella spiralis. Metformin and atorvastatin are applied on a wide scale, to the degree that they could be considered as part of the host biochemical environment that can affect the parasite. Therefore, this study aimed to investigate the impact of alteration of the host's biochemical environment by these commonly used drugs upon the course of T. spiralis infection. Mice were divided into three groups: (1) received atorvastatin, (2) received metformin, and (3) untreated, then after one week, animals were infected with T. spiralis. The treatment continued until the end of the experiment. From each group, small intestines and muscles were removed for histopathological, immunohistochemical, and biochemical analyses as well as total muscle larval counts. We found that the oxidative stress and the expression of vascular endothelial growth factor (VEGF) in the muscles were significantly reduced in both drug-receiving groups, while the total larval counts in muscles were only significantly reduced in atorvastatin-receiving group as compared to the infected control group. Moreover, marked reduction in the inflammatory cellular infiltration, cyclooxygenase-2 (COX-2) expression, and oxidative stress was noted in the small intestines of the treated groups as compared to the infected control group. In conclusion, this study provides many insights into the different biochemical changes in the host that the parasite has to face. Moreover, the anti-inflammatory and anti-angiogenic effects should be taken into consideration when treating infections in patients on therapy with atorvastatin or metformin.

Immunopathological changes in the brain of immunosuppressed mice experimentally infected with Toxocara canis.

Toxocariasis is a soil-transmitted helminthozoonosis due to infection of humans by larvae of Toxocara canis. The disease could produce cognitive and behavioral disturbances especially in children. Meanwhile, in our modern era, the incidence of immunosuppression has been progressively increasing due to increased incidence of malignancy as well as increased use of immunosuppressive agents. The present study aimed at comparing some of the pathological and immunological alterations in the brain of normal and immunosuppressed mice experimentally infected with T. canis. Therefore, 180 Swiss albino mice were divided into 4 groups including normal (control) group, immunocompetent T. canis-infected group, immunosuppressed group (control), and immunosuppressed infected group. Infected mice were subjected to larval counts in the brain, and the brains from all mice were assessed for histopathological changes, astrogliosis, and IL-5 mRNA expression levels in brain tissues. The results showed that under immunosuppression, there were significant increase in brain larval counts, significant enhancement of reactive gliosis, and significant reduction in IL-5 mRNA expression. All these changes were maximal in the chronic stage of infection. In conclusion, the immunopathological alterations in the brains of infected animals were progressive over time, and were exaggerated under the effect of immunosuppression as did the intensity of cerebral infection.

Effect of propranolol on IL-10, visfatin, Hsp70, iNOS, TLR2, and survivin in amelioration of tumor progression and survival in Solid Ehrlich Carcinoma-bearing mice.

BACKGROUND: ß-Adrenergic signaling could contribute to initiation and progression of breast cancer. This research investigated some potential mechanisms of propranolol in amelioration of progression and survival in breast cancer. METHODS AND RESULTS: Solid Ehrlich Carcinoma (SEC) xenograft model was induced in 30 mice divided into 3 groups; where group I served as untreated SEC group. In groups II and III, propranolol treatment i.p. in low (5mg/kg) and high dose (10mg/kg) caused significant increase in interleukin-10 (IL-10) and decrease in heat shock protein 70 (Hsp70) and inducible nitric oxide synthase (iNOS) activity with non significant change in visfatin in tumor tissues compared to untreated SEC. In untreated SEC, tumor volume (V) exhibited significant negative correlation with IL-10 levels and toll like receptor 2 (TLR2) expression with significant positive correlation with Hsp70 levels and iNOS activity. While propranolol in either doses caused reduction of tumor volume (V), and improved percentage tumor growth inhibition (% TGI) only its high dose exhibited significant impact on survival rate. Propranolol dose-dependent effect was evident for IL-10 and Hsp70, and even only the high dose significantly increased and decreased TLR2 and survivin, respectively. This comes in favor of recommending high dose of propranolol in cancer therapy. Nonetheless, use of low dose cannot be ignored when benefit to risk balance have to be considered. CONCLUSIONS: Propranolol could provide palliative effects in progression and survival of breast cancer that are mainly mediated via direct immunomodulatory and apoptotic mechanisms and probably associated with indirect anti-angiogenic activity.

Biochemical and immunopathological changes in experimental neurotoxocariasis.

Toxocariasis is a widespread soil-transmitted parasitic disease. Toxocara canis larvae migrate through the tissues with a special predilection for the central nervous system. Recently, neurotoxocariasis is being diagnosed in humans with increasing frequency due to improved diagnostic tools. The present study aimed at exploring the biochemical and immunopathological alterations in the brain in experimental T. canis infection. For this purpose, 75 Toxocara-infected mice were sacrificed at 2, 5, and 16 weeks post-infection. The brains were removed and assayed for total larval count, pro-inflammatory cytokines (TNF-alpha, IL-6), and central neurotransmitters (gamma-aminobutyric acid, glutamate, dopamine, norepinephrine, and serotonin). Brain sections were also stained for histopathological study, and for assessment of the expression of inducible nitric oxide synthase (iNOS), and glial fibrillary acidic protein (GFAP) by immunohistochemical methods. We found that larval recovery showed progressive increase over the course of infection. Furthermore, the infected mice displayed increased expression of pro-inflammatory cytokines and iNOS, as well as significant disturbances in neurotransmitter profile. Astrocytic activation, evidenced by enhanced expression of GFAP, was also manifest in infected animals. These changes were maximal in the chronic stage of infection or intensified over time. In conclusion, experimental neurotoxocariasis is associated with significant biochemical, immunological, and pathological changes.

Congenital exposure to Schistosoma mansoni infection: impact on the future immune response and the disease outcome.

Schistosomiasis mansoni is a widespread parasitic infection that may lead to several serious complications, such as hepatic periportal fibrosis and portal hypertension, mainly due to deposition of schistosome eggs in the tissues. However, people in endemic areas infrequently exhibit severe pathology and complications; this may be explained, in part, by modulation of the disease in indigenous populations by in utero exposure to the parasite. This study investigated the differences between mice born to Schistosoma mansoni-infected mothers and those born to non-infected ones in subsequent postnatal schistosomal infections. We found that the intensity of infection, evidenced by hepatic egg load, was much reduced in mice born to infected mothers. No difference was found as regards total and Schistosoma-specific immunoglobulin levels except for total IgG. The levels of gene expression of two regulatory cytokines, namely interleukin-12 (IL-12) and transforming growth factor beta (TGF-beta) were found to be significantly increased in prenatally exposed animals. Moreover, liver fibrosis was significantly decreased in animals born to infected mothers as revealed by histopathological and histochemical examination as well as by immunohistochemical identification of activated hepatic stellate cells (HSCs) using antibody against glial fibrillary acidic protein (GFAP). In conclusion, congenital exposure to S. mansoni seems to ameliorate the immunopathological changes in future postnatal infections.

Immunohistochemical expression of topoisomerase II alpha and Her-2/neu in prostatic carcinoma and benign prostatic hyperplasia.

BACKGROUND AND PURPOSE: Topoisomerase II a (Topo II a) and Her-2/neu are two important targeted therapeutic molecules. The immunohistochemical expression of both of them has not been widely studied in prostatic carcinoma and benign prostatic hyperplasia (BPH). The aim of this study was to evaluate the immunohistochemical expression of Topo II a and Her-2/neu in prostatic carcinoma and BPH and compare the expression patterns of both genes in cases of prostatic carcinoma in relation to Gleason score and hormonal status. MATERIAL AND METHODS: Paraffin blocks of 30 cases of prostatic carcinoma (categorized by Gleason score and hormonal status) and 5 cases of BPH presented to the Department of Pathology, Faculty of Medicine, Tanta University during the period from 2005 to 2008 were retrieved from the files. The immunohistochemical expression of Topo II a and Her-2/neu antibodies in the above-mentioned diagnostic categories was investigated and compared. The percentage of nuclei staining for Topo II a was semiquantitated; overexpression was defined as >or=5% nuclear staining. Her-2/neu immunoreactivity was scored from 0 to 3 + depending on membrane staining intensity and pattern. RESULTS: The expression of Topo II a varied significantly among the different studied groups (p<0.001). Topo II a expression increased significantly with increased Gleason score in prostatic carcinoma (p=0.001). Its expression in both moderately and poorly differentiated carcinomas was significantly higher than in BPH (p=0.005 and 0.002 respectively); however the difference between its expression in well-differentiated carcinoma and in BPH was statistically insignificant (p=0.171). Her-2/neu expression was higher in prostatic carcinoma than in BPH, however the difference did not reach the level of statistical significance (p=0.084). Also, the increase in its expression within prostatic carcinoma cases with increased Gleason score was statistically insignificant (p=0.100). There was a significant correlation between Topo II a and Her-2/neu expression (p=0.008, r=0.478). Hormone resistant carcinomas showed higher expression of Topo II a and Her- 2/neu than carcinomas with no hormone treatment, however, the differences were statistically insignificant (p=0.594 and 0.667 respectively). CONCLUSIONS: Topo II a expression was significantly higher in poorly differentiated and moderately differentiated prostatic carcinoma compared to BPH. There was a significant increase in Topo II a expression with increased Gleason score. Her-2/neu expression was higher in prostatic carcinoma than in BPH, however the difference did not reach the level of statistical significance and the increase in its expression with increased Gleason score was also statistically insignificant. Topo II a and Her-2/neu were co-expressed significantly. Hormone resistant carcinomas showed higher expression of both markers, however, the differences were statistically insignificant. The latter finding may have important therapeutic implications, however, further large scale studies are required for confirmation. KEYWORDS: Topo II a - Her-2/neu - Prostatic carcinoma - BPH.

P63 and cytokeratin8/18 expression in breast, atypical ductal hyperplasia, ductal carcinoma in situ and invasive duct carcinoma.

Background and Purpose : The pattern and distribution of p63 expression as a myoepithelial/basal stem cell marker can be different between atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) and may denote basal phenotype of breast ductal carcinoma. CK8/18 is a luminal marker and may indicate a luminal phenotype of IDC and its expression in ADH and DCIS may refer to a possible precursor lesion to IDC. This work was designed to study and compare the expression of p63 and cytokeratin 8/18 (CK8/18) in some cases of ADH, DCIS and IDC. Materials and Methods : Histopathological evaluation and immunohistochemical study of anti-p63 and anti- CK8/18 was performed on selected archival cases of 7 ADH, 12 DCIS, 30 IDC of known clinicopathological data and previous estrogen receptor status (ER) for IDC. Confirmatory anti-smooth muscle actin (ASMA) expression for positive p63 cases was performed. Results : p63 was expressed in the peripheral rim of the myoepithelial cell layer in ADH and DCIS with occasional gabs in DCIS. It was positive and stained occasional malignant cells in 3/30 (10%) of IDC cases. Confirmatory ASMA staining decorated the same peripheral rim of cells in ADH and DCIS, but was negative in p63 positive IDC cases. CK8/18 was positive in 100% of ADH, 8/12 (66.7%) of DCIS and 22/30 (73%) of IDC cases. Combined p63 and CK8/18 expression was noticed in 3/30 (10%) of IDC. Conclusion : It is concluded from this study that p63 is specific and valuable in differentiating myoepithelial cells and is more specific and valuable than other myoepithelial markers, as ASMA and can differentiate between ADH, DCIS, IDC as it stains peripheral myoepithelial cells in ADH and DCIS with gabs in the latter and does not stain any neoplastic cells. In IDC, it is positive in malignant cells in a minority of cases which may indicate basal/stem cell/myoepithelial cell origin of breast carcinoma. Comparatively, CK8/18 cannot differentiate ADH, DCIS and IDC as there is no difference in its staining pattern among them, which may suggest that they are a continuum or that ADH and DCIS are precursors for the luminal phenotype of IDC. Key Words : p63 -CK8/18 -IDC -ADH -DCIS -Basal/ stem cells.