Restoration of renal hemodynamics and functions by Nigella sativa administration in dinitrophenol-induced hypoxia in rat's animal model.

Objective: Hypoxia is one of the principal causes of renal diseases. This study aimed to evaluate the effects of Nigella sativa on dinitrophenol (DNP)-induced hypoxia renal damage in rats. Methods: Forty adult male rats were incorporated in this study. The rats were divided into four groups: control group, N. sativa group, DNP hypoxic group, and DNP + N. sativa group receiving N. sativa (400 mg/kg body weight). Serum and renal tissue erythropoietin (EPO) hormone and hypoxia-inducible factor-2α (HIF-2α) levels were measured. Renal oxidative stress biomarkers, inflammatory biomarkers, renal hemodynamics, and histopathological examination were evaluated. Results: Administration of N. sativa highly significantly normalized serum EPO level, HIF-2α (P < 0.001 for each) in DNP + N. sativa treated rats as compared to DNP hypoxic rats. Furthermore, it highly significantly improved renal oxidative stress evident by decreased renal tissues malondialdehyde and increased superoxide dismutase, total thiol, and catalase activity (P < 0.001 for each). Furthermore, a highly significant decline of renal intercellular adhesion molecule-1, myeloperoxidase, and interleukin-6 was observed in DNP + N. sativa rats (P < 0.001 for each). Improvements in renal hemodynamics and kidney functions were also found after N. sativa administration (with P < 0.001 for all parameters). In addition, N. sativa treatment reduced renal histopathological changes of the DNP + N. sativa group. Our results were statistically analyzed using the Prism software package (GraphPad version 8.0). Conclusion: N. sativa has an alleviating effect on DNP-induced hypoxia renal damage and can restore kidney functions in rats' animal models. These effects were through antioxidant, anti-inflammatory, and hemodynamic mechanisms.

FOS-Like Antigen 1 Expression Was Associated With Survival of Hepatocellular Carcinoma Patients.

Background: Early diagnosis and proper management of hepatocellular carcinoma (HCC) improve patient prognosis. Several studies attempted to discover new genes to understand the pathogenesis and identify the prognostic and predictive factors in HCC patients, to improve patient's overall survival (OS) and maintain their physical and social activity. The transcription factor FOS-like antigen 1 (FOSL1) acts as one of the important prognostic factors in different tumors, and its overexpression correlates with tumors' progression and worse patient survival. However, its expression and molecular mechanisms underlying its dysregulation in human HCC remain poorly understood. Our study was conducted to evaluate the expression of FOSL1 in HCC tissues and its relationship with various clinicopathological parameters besides OS. Methods: This study is a retrospective cohort study conducted among 113 patients with a proven diagnosis of HCC, who underwent tumor resection and received treatment at South Egypt Cancer Institute. Immunohistochemistry for FOSL1 expression and survival curves were conducted followed by statistical analysis. Results: HCC occurred at older age group and affected males more than females. There was a statistically significant correlation between combined cytoplasmic and nuclear expression of FOSL1 and worse prognosis in HCC patients. There was a statistically significant correlation of FOSL1 expression with histological grade, lymphovascular embolization, and tumor budding where high expression indicated potential deterioration of HCC patients. There was statistically significant correlation between tumor size, tumor grade and FOSL1 expression with the cumulative OS. Conclusions: Combined cytoplasmic and nuclear FOSL1 expression has significant prognostic association with HCC and diagnostic importance, as it can identify cirrhosis and premalignant lesions that can progress to HCC. Furthermore, Kaplan-Meier survival analysis found that overexpressed FOSL1 was correlated with poor OS.

Absence of CD74 Isoform at 41kDa Prevents the Heterotypic Associations between CD74 and CD44 in Human Lung Adenocarcinoma-derived Cells.

Lung cancer is a leading cause of cancer-associated death in all over the globe. This study was undertaken to determine the expression and interaction of membrane-bound receptors CD74 and CD44 in human lung adenocarcinoma cells and their associated signaling was also attempted. Levels of CD74 and CD44 were studied in human lung adenocarcinoma-evolved cells A549 and H460. CD74-mediated downstream signaling was studied by the nuclear-transcription-factor NF-κB and prostaglandin E2 (PGE2) production. Flow-cytometric analysis showed that both CD74 and CD44 were perfectly expressed in A549 cells. Importantly, Western immunoblotting showed that A549 cells expressed only two isoforms of CD74 at 33 and 35 kDa but isoform at 41 kDa was absent. These results were verified in H460 cells. Confocal microscopy showed CD74 and CD44 was colocalized but heterotypic interaction between them was missing in both A549 and H460 cells. Activation of NF-κB and production of PGE2 in human lung cancer cells were comparable with other cancer cells. In conclusion, this is the first study that shows A549 and H460 cells expressed two distinctive isoforms of CD74 but isoform at 41 kDa was absent. Due to the absence of this isoform, the direct physical interaction between them CD74 and CD44 was lacking. Furthermore, the data also demonstrated that lacking of direct physical interaction between CD74 and CD44 had no effect on NF-κB activation and PGE2 production indicating that CD74-mediated downstream signaling occurs either through coreceptors or indirect interaction with CD44 in human lung cancer cells.Abbreviation: CD: cluster of differentiation; SCLC: small cell lung cancer; NSCLC: nonsmall cell lung cancer; SCC: squamous cell carcinoma; ADC: adenocarcinoma; LCC: large cell carcinoma.

EGFR and CXCR1 expression in thyroid carcinoma in Qassim Region-Saudi Arabia: Correlation with clinicopathological parameters.

AIMS: Recent evidence indicates an increased incidence of thyroid carcinoma, especially papillary thyroid carcinoma (PTC), in Saudi Arabia. EGFR and CXCR1 were reported to have increased expression in several human neoplasms. The goals of the present research was to investigate EGFR and CXCR1 expression in thyroid carcinoma and correlate the results to the established prognostic factors. METHODS: Immunohistochemical study for both EGFR and CXCR1 was performed on formalin-fixed paraffin-embedded thyroid carcinomas tissues sections applying Labeled Streptavidin-biotin method (LSAB). RESULTS: Remarkable high expression of EGFR and CXCR1 were observed in PTC cases (56% and 63% respectively). There was association between EGFR expression in PTC and each of histologic subtype, lymph node metastasis (LNM), distant metastasis (DM), TNM staging and tumor relapse. There was statistical significant correlation between CXCR1 expression in PTC and each of histologic subtype, LNM, and tumor relapse. A significant correlation was detected between concomitant EGFR and CXCR expression and LNM, DM, increasing stage and tumor relapse. CONCLUSIONS: The results of the present study demonstrated, a statistically positive correlation of EGFR and CXCR1 expression in PTC compared to normal thyroid tissues and nodular hyperplasia in Qassim Region- Saudi Arabia. Concomitant high expression of both receptors were strongly correlated with LNM, DM, TNM stage and tumor relapse than did each alone. These findings suggest that EGFR and CXCR1 play crucial roles in PTC and serve as predictors of poor prognosis, biomarkers of tumor diagnosis, and potential targets of cancer therapeutics.

Cytoskeletal Focal Adhesion Proteins Fascin-1 and Paxillin Are Predictors of Malignant Progression and Poor Prognosis in Human Breast Cancer.

Breast cancer is a major health problem in both developing and developed countries. The incremental motility of malignant cells is a critical step in their migration, invasion, and metastasis and is regulated by the reorganization of the actin cytoskeleton and regulation of focal adhesion. Fascin-1 and paxillin are essential components of these cellular structures. In cancer, the expression level of fascin-1 and paxillin can vary depending on cell type. However, its precise role in breast cancer of Saudi women has not been evaluated in any published study. We investigated fascin-1 and paxillin expression in breast carcinoma, and we have related these results to the established prognostic factors. We studied 100 breast carcinoma specimens. Immunohistochemical analyses for fascin-1 and paxillin were conducted on paraffin sections of breast tissues using the avidin-biotin peroxidase method. Fascin-1 and paxillin were expressed in 58% and 43% of infiltrating duct carcinoma (IDC) cases. There was a statistically significant correlation between fascin-1 and paxillin expression with each of the following: tumor grade, clinical stage, lymph-node metastasis grade, and HER2 expression. Furthermore, there was a significant correlation between fascin-1 expression with paxillin immunostaining but no such association with PR or ER status. Increased fascin-1 and paxillin expression in IDC cells and their correlation with poor prognostic factors support their strong correlation with tumor progression, invasion, and metastasis in human breast cancer, indicating that these markers can be used as a target for the development of novel therapies.

The effects of alpha-lipoic acid on breast of female albino rats exposed to malathion: Histopathological and immunohistochemical study.

BACKGROUND: The wide use of the organophosphate insecticide malathion is accompanied by the risk of human exposure and may be involved in the etiology of breast cancers, especially in developing countries. Alpha (α)-lipoic acid, a natural molecule, present in our diet has antioxidant and protective effects in cases such as aging, diabetes mellitus, and vascular and neurodegenerative diseases all in which free radicals are involved. However, there is only scarce data regarding the efficacy and biological activity of α-lipoic acid on malathion-induced breast histopathological changes. AIMS: To investigate whether malathion can induce mammary histopathological changes, to immunohistochemically analyze the modulations in proliferation-apoptosis balance associated with these changes, to assess the associated metabolic parameters, antioxidant stress and hormonal profile changes and to elucidate the possible protective effect of α-lipoic acid on malathion induced alterations in rats. MATERIALS AND METHODS: Forty Wistar female rats weighing 150-170g were divided into four groups. Group 1: control group were injected intraperitoneally (ip) with saline solution. Group2: animals were injected (ip) with malathion twice a day for five days. Group 3: animals were orally given α-lipoic acid, after three hours of treatment with malathion at the same dose given to group 2. Group 4: animals were treated with α-lipoic acid at the same dose given to group 3. Rats were sacrificed on the 90th day, and breast tissues were analyzed for histopathological and immunohistochemical alterations. Blood samples were collected for biochemical tests. RESULTS: α-Lipoic acid exhibited a striking reduction of malathion-induced mammary tumor incidence, and reversed intra-tumor histopathological alterations. Alpha lipoic acid suppressed proliferating cell nuclear antigen (PCNA) and p53 expression, induced apoptosis, upregulated proapoptotic protein Bax. CONCLUSIONS: Our results provide the experimental evidence that α-lipoic acid exerts chemopreventive effect in the breast hyperplastic and malignant changes by suppressing abnormal cell proliferation and inducing apoptosis with an oncostatic effects during an early-stage breast cancer.

Expression of tumor necrosis factor-related apoptosis-inducing ligand death receptors DR4 and DR5 in human nonmelanoma skin cancer.

Death receptors 4 and 5 (DR4 and DR5) are cell surface receptors that when activated by their ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in most cancer cells but not in normal cells. Currently, it remains unclear whether DR4 and DR5 are involved in immune surveillance against nonmelanoma skin cancer (NMSC) progression. The aim of this study was to investigate the expression of DR4 and DR5 in NMSC and relate the results to the established clinicopathologic prognostic factors. This study was conducted on about 80 skin specimens from patients with NMSC (40 basal cell carcinoma and 40 squamous cell carcinoma) and diagnosed and confirmed by biopsy. Immunohistochemical analysis for DR4 and DR5 was carried out on formalin-fixed paraffin-embedded sections of skin tissues using avidin-biotin peroxidase method. Significant expression of both DR4 and DR5 was observed in NMSC cases. There was statistically significant association between DR4 and DR5 expression in squamous cell carcinoma and each of tumor site and lymph node metastasis. There was statistically significant association between DR4 expression in basal cell carcinoma and histopathologic subtypes (high expression in nodular type) and between DR5 expression and tumor site (high expression in sun-exposed area). In conclusion, expression of TRAIL receptors that mediate extrinsic apoptotic pathway in NMSC may be suggestive of a reassessment of the suitability of TRAIL-based strategy in future NMSC therapies.

Effects of thymoquinone on STZ-induced diabetic nephropathy: an immunohistochemical study.

BACKGROUND: Thymoquinone (TQ) is the most abundant and active ingredient of Nigella sativa (NS) seeds. Its hepatic, renal, and cardiac protective effects have been demonstrated in animal models. Streptozotocin (STZ) is an antibiotic that is widely used experimentally as an agent capable of inducing insulin-dependent diabetes mellitus (IDDM), also known as type I diabetes mellitus (T1DM). OBJECTIVES: This study was carried out in an attempt to highlight the possible beneficial effects of TQ in STZ-induced diabetes in rats and to determine the predictive value of mesenchymal and epithelial markers in the response of diabetic nephropathy to TQ. MATERIALS AND METHODS: Sixty adult male albino rats were divided in 3 groups: control, diabetic untreated, and diabetic treated with TQ. RESULTS: Diabetic rats exhibited morphological changes in both renal glomeruli and tubules with immunohistochemical expression of the mesenchymal markers Fsp1, desmin, and MMP-17 and disappearance of the epithelial marker ZO-1 largely in the glomeruli of diabetic kidneys. Treatment with TQ significantly attenuated renal morphological and immunohistochemical changes in STZ-induced diabetic rats. CONCLUSIONS: Thymoquinone has protective effects on experimental diabetic nephropathy. Both mesenchymal and epithelial markers serve as excellent predictors of early kidney damage and indicators of TQ responsiveness in STZ-induced diabetic nephropathy.

The role of heat shock protein 70 induced by geranylgeranylacetone in carbon tetrachloride-exposed adult rat testes.

Carbon tetrachloride (CCl4) induces testicular damage, through formation of free-radical metabolites. Molecular chaperone heat shock protein of 70kDa (HSP 70) protects cells from various stresses. This study was designed to investigate the potential role of induction of HSP70 using geranylgeranylacetone (GGA) on testicular damage caused by CCl4. Rats were divided into group I (control group), Group II (CCl4 group) received CCl4 s.c. for 4 weeks, group III received CCl4 s.c. for 4 weeks simultaneously with daily single oral dose of GGA (GGA - treated CCl4 group). Serum testosterone, testicular lactate dehydrogenase (LDH) and alkaline phosphatase (ALP), testicular malondialdehyde (MDA), total nitrite and total antioxidant capacity (T-AOC) were measured. Evaluation of histopathological changes and immunohistochemical HSP70 expression for testicular biopsies were performed. Group II showed lower values of gonado-somatic index, serum testosterone, testicular LDH, ALP, T-AOC and greater values of testicular MDA and total nitrite than in control. Testicular morphology showed widening of seminiferous lumen, less spermatogenesis, vacuolization of germinative epithelium. Group III had higher values of gonado-somatic index, serum testosterone, testicular LDH, ALP, T-AOC with less testicular MDA and total nitrite than in group II. They have less damage and restored the altered testicular morphology. Immunohistochemical HSP70 expression was increased in the testicular spermatogenic and sertoli cells in group II that was significantly accentuated in group III. These findings suggest that GGA-induced activation of HSP 70 significantly alleviate CCl4 inflicting testicular damage by HSP 70 mediated cytoprotection and antioxidant effects.

Histopathological study of evening primrose oil effects on experimental diabetic neuropathy.

UNLABELLED: Diabetic polyneuropathy is a serious complication of diabetes mellitus and the most frequent neuropathy worldwide. AIM: This study was designed to investigate the possible beneficial effects of evening primrose oil (EPO) on histopathological changes of sciatic nerves in streptozotocin-induced diabetic rats. MATERIALS AND METHODS: The rats were randomly allotted into three experimental groups: A (control), B (diabetic untreated), and C (diabetic treated with EPO); each group contained 10 animals. Groups B and C received streptozotocin (STZ) to induce diabetes. The rats in group C were given EPO for 2 weeks after 6 weeks of STZ injection. Blood and tissue samples were obtained for biochemical and histopathological investigation. RESULTS: STZ-treated diabetic rats showed reduction of the size of islets of Langerhans, fatty degeneration in the pancreatic acini with dilation, irregularity, and increased thickness of blood vessels. Electron micrography of sciatic nerves of diabetic rats showed multiple vaculations and partial separation of myelinated nerve fibers with axonal atrophy, endoneural edema, and increased collagen fibers. Compared with diabetic rats, EPO induced partial recovery from diabetes-induced pancreatic and nerve damage. CONCLUSIONS: Histologic evaluation of the tissues in diabetic animals treated with EPO showed fewer morphologic alterations with significant decrease of myelin breakdown. Furthermore, the ultrastructural features of axons showed partial improvement. It is believed that further preclinical research into the utility of EPO may indicate its usefulness as a potential treatment on peripheral neuropathy in STZ-induced diabetic rats.

CD44s and CD44v6 in diagnosis and prognosis of human bladder cancer.

UNLABELLED: The cell adhesion molecules (CAMs) CD44 standard (CD44s) and its variant 6 (CD44v6) are involved in the progression and invasion of human malignancies. However, discrepancies in the prognostic value of CD44s and CD44v6 expression need to be addressed. AIMS: To investigate the expression of CD44s and CD44v6 in bladder carcinomas and relate the results to the established prognostic factors. MATERIALS AND METHODS: 50 bladder carcinoma specimens, 30 cases with transitional cell carcinoma (TCC: 6 bilharzial and 24 nonbilharzial) and 20 cases with squamous cell carcinoma (SCC: 8 bilharzial and 12 nonbilharzial), were included. Immunohistochemical analysis for CD44s and CD44v6 was carried out using avidin-biotin peroxidase method. RESULTS: The level of both CD44s and CD44v6 in TCC was significantly higher in invasive than in preinvasive tumors and normal urothelium (p < .05). A direct association between the percentage of expression of both markers and the grade of TCC (p < .05) was observed. An inverse correlation between CD44s and SCC was seen, where metaplastic urothelium showed higher expression than invasive carcinomas. No association was observed between the expressions of both CD44s and CD44v6 and bilharzial ova, sex and age of the patient, or size of the tumor. CONCLUSIONS: The authors report statistically significant correlation between CD44s and CD44v6 expression and increasing grade and stage of TCC. No such correlation with SCC and with bilharzial cystitis, sex and age of the patient, or size of the tumor was documented.

Gelatinases A and B expression in human colorectal cancer in upper Egypt: a clinicopathological study.

AIM: Prognosis of colorectal carcinoma depends on many factors, such as age and sex of patient; location; multiplicity; local extent and size of tumor, bowel obstruction, or perforation; as well as tumor microscopic type and grade; vascular and perineural invasion; and nodal and distant metastasis. The matrix metalloproteinases (MMPs) are a family of proteolytic enzymes strongly implicated in tumor invasion and metastasis, hence in tumor prognosis. The purpose of this study was to assess the role of MMP-2 and MMP-9 expression in colorectal tumorigenesis, invasion, and metastasis, hence their prognostic values. METHOD: Immunohistochemical analysis of MMP-2 and MMP-9 in colorectal cancer cells, an immunohistochemical score based on the intensity of immunoreactivity and proportion of immunoreactive cells that established for each MMP, and correlation of this expression with the established prognostic factors. RESULTS: MMP-2 was expressed in 81.8% (strong expression in 40%) of cases, and MMP-9 was expressed in 72% (strong expression in 35%) of cases. CONCLUSIONS: MMP-2 and MMP-9 are widely expressed in colorectal carcinoma, suggesting significant diagnostic and prognostic values in these tumors. Increased levels of MMP-2 and MMP-9 protein expression in colorectal carcinoma tissues as compared to normal tissues suggest their association with colorectal tumor invasion and metastasis and that they could be targets for intervention and therapy in colorectal carcinoma.

The prognostic value of breast cancer resistance protein (BCRB/ABCG2) expression in breast carcinomas.

In cancer, the level of expression of breast cancer resistance protein (BCRP/ABCG2) can vary depending on cell type. However, its precise role in breast cancer has been controversial. The aim of this study was to investigate the expression of ABCG2 in breast carcinomas and relate the results to the established prognostic factors. An immunohistochemical study was conducted on 200 breast carcinoma specimens using the avidin-biotin peroxidase method. ABCG2 was expressed in 77% of cases of invasive ductal carcinoma. There was a statistically significant correlation between ABCG2 expression and each of the tumor grade, clinical stage, lymph node metastasis, and HER2 immunostaining, but no such association with progesterone receptor (PR) and estrogen receptor (ER) status. Increased expression of ABCG2 in invasive ductal carcinoma cells and its statistically significant correlation with HER2 expression are strongly correlated with tumor progression, invasion, and metastasis in human breast cancer and indicates that ABCG2 can be used as a target for the development of novel therapies.

CD10 and E-cad expression in urinary bladder urothelial and squamous cell carcinoma.

Identification of prognostic markers in bladder carcinoma comprises a major clinical issue and therapeutic target. CD10 and the adhesion molecule E-cadherin (E-cad) are expressed in a variety of normal tissues and their neoplasms. CD10 is able to degrade extracellular matrix and other proteins, including adhesion molecules. The roles of CD10 and E-cad and their relationship in the development and progression of bladder carcinoma are poorly understood. The aim of this study was to investigate the expression of CD10 and E-cad in bladder carcinomas and relate the results to the established prognostic factors. This study included 144 patients with bladder carcinoma, including 72 with transitional cell carcinoma (TCC; 30 bilharzial and 42 nonbilharzial) and 72 with squamous cell carcinoma (SCC; 38 bilharzial and 34 nonbilharzial). Immunohistochemical analysis for both CD10 and E-cad were carried out on paraffin-fixed sections of neoplastic bladder tissues. CD10 tumor cells, CD10 stromal cells, and E-cad were expressed in 56%, 58%, and 51% of cancer bladder cases, respectively. There was a statistically significant correlation between percentage of tumor cells positively stained by CD10 and each of the tumor grade, clinical stage, and lymph node metastasis of both TCC and SCC. There was a significant statistical correlation between immunostaining by E-cad marker and each of the tumor grade, clinical stage, and lymph node metastasis of TCC, but no such association with SCC. The frequency of stromal expression of CD10 was higher in bilharzial-associated bladder carcinomas than in nonbilharzial ones, and these results were statistically significant. In conclusion, increased expression of CD10 in the tumor and stromal cells of both TCC and SCC and decreased expression of E-cad in the tumor cells of only TCC are strongly correlated with tumor progression, invasion, and metastasis in human bladder cancer.

Expression of matrix metalloproteinases 2 and 9 in human trophoblasts of normal and preeclamptic placentas: preliminary findings.

OBJECTIVE: Here we test the hypothesis that "the expression of matrix metalloproteinase 2 and 9 proteins is altered in preeclamptic placentas compared to placentas of normal pregnancy." PATIENTS AND METHODS: This case-control study includes preeclamptic placentas (40 women with preeclampsia) from a singleton pregnancy and placentas of normal pregnancies (control group, 40 women with uncomplicated pregnancy). The expression patterns of metalloproteinases 2 and 9 were examined using immunohistochemical staining methods. RESULTS: Compared to uncomplicated pregnancy, the incidence of intrauterine growth restriction was high and the mean birth weight was markedly low in patients with preeclampsia. Both metalloproteinase 2 and 9 proteins were frequently and strongly expressed in the majority of placentas of uncomplicated pregnancies (control group). Metalloproteinase 9 expression was absent in the majority of the preeclamptic placentas. In the remaining cases of preeclamptic placentas, the expression of metalloproteinase 9 was weak. In contrast, a strong metalloproteinase 2 protein expression was seen in the majority of the preeclamptic placentas. CONCLUSIONS: These preliminary data demonstrate the expression of metalloproteinase 2 and 9 proteins in the placentas of uncomplicated pregnancies. The absence/reduced expression of metalloproteinase 9 in the preeclamptic placentas may be related to insufficient invasion of trophoblast, leading to superficial and unsuccessful placentation.