Clinical features and outcomes of infective endocarditis in Egypt: an 11-year experience at a tertiary care facility.

BACKGROUND: Few data are available on the characteristics of infective endocarditis (IE) cases in Egypt. The aim of this work is to describe the characteristics and outcomes of IE patients and evaluate the temporal changes in IE diagnostic and therapeutic aspects over 11 years. RESULTS: The IE registry included 398 patients referred to the Endocarditis Unit of a tertiary care facility with the diagnosis of possible or definite IE. Patients were recruited over two periods; period 1 (n = 237, 59.5%) from February 2005 to December 2011 and period 2 (n = 161, 40.5%) from January 2012 to September 2016. An electronic database was constructed to include information on patients' clinical and microbiological characteristics as well as complications and mortality. The median age was 30 years and rheumatic valvular heart disease was the commonest underlying cardiac disease (34.7%). Healthcare-associated IE affected 185 patients (46.5%) and 275 patients (69.1%) had negative blood cultures. The most common complications were heart failure (n = 148, 37.2%), peripheral embolization (n = 133, 33.4%), and severe sepsis (n = 100, 25.1%). In-hospital mortality occurred in 108 patients (27.1%). Period 2 was characterized by a higher prevalence of injection drug use-associated IE (15.5% vs. 7.2%, p = 0.008), a higher staphylococcal IE (50.0% vs. 35.7%, p = 0.038), lower complications (31.1% vs. 45.1%, p = 0.005), and a lower in-hospital mortality (19.9% vs. 32.1%, p = 0.007). CONCLUSION: This Egyptian registry showed high rates of culture-negative IE, complications, and in-hospital mortality in a largely young population of patients. Improvements were noted in the rates of complications and mortality in the second half of the reporting period.

Impact of Mesenchymal Stem Cells and Vitamin D on Transforming Growth Factor Beta Signaling Pathway in Hepatocellular Carcinoma in Rats

Background: Transforming growth factor-beta (TGF-ß) signaling is recognized as being critical for carcinogenesis. Vitamin D has proved to exert numerous tumor suppressive effects. Effects of bone marrow derived mesenchymal stem cells (BM-MSCs) on tumor progression are still controversial. The present study was conducted to evaluate the effects of BM-MSCs and vitamin D on TGF-ß signaling in an experimental hepatocellular carcinoma (HCC) model in rats. Materials and Methods: The study was conducted on fifty female white albino rats divided equally into 5 groups: controls, HCC induced by diethyl-nitrosamine (DENA) and carbon tetrachloride (CCl4), HCC plus MSCs, HCC plus vitamin D and HCC plus both MSCs and vitamin D. The following parameters were assessed in rat liver tissues: TGF-ß and Smad2 protein levels by ELISA and western blotting, respectively, gene expression of Smad3, Smad7, Snail, HNF4α and MMP-2 and histopathological lesions. Serum levels of alpha fetoprotein (AFP), ALT and albumin were also assessed. Results: TGF-ß protein levels and gene expression of its downstream effectors (Smad3 and Snail), in addition to Smad2 protein levels were significantly higher in the HCC group than in the control group. On the other hand, they were significantly down-regulated in all treated groups with most significant amelioration with both MSCs and vitamin D. Also, the serum levels of AFP were significantly increased in the untreated HCC group, and this was again reversed in all treated groups. Histopathological examination of liver tissue revealed that administration of MSCs or vitamin D into HCC rat group improved the histopathological picture with residual tumor pathology, while administration of both MSCs and vitamin D showed better restoration of liver parenchyma. These data suggest that the TGF-ß signaling pathway could be used as a therapeutic target in HCC.

Mesenchymal stem cell therapy of hepatocellular carcinoma in rats: Detection of cell homing and tumor mass by magnetic resonance imaging using iron oxide nanoparticles.

BACKGROUND: Bone marrow-derived mesenchymal stem cells (MSCs) are reported to improve hepatic fibrosis, and may impact the signaling mechanisms leading to the induction of hepatocellular carcinoma (HCC) in animal models of liver cirrhosis. OBJECTIVES: The aim of this study was to clarify and explain the therapeutic role played by MSCs in hepatic cirrhosis and HCC by tracking them using nanoparticles. MATERIAL AND METHODS: Liver cirrhosis and HCC were established in rats with the use of carbon tetrachloride and diethylnitrosamine injection. Magnetic resonance imaging (MRI) was used to track nanoparticlelabeled MSCs in the intact animal following injection and to monitor the changes in the hepatic parenchyma. RESULTS: Labeling of MSCs with iron oxide nanoparticles did not adversely affect their viability and proliferation. MRI indicated a significant reduction in tumor mass in the labeled MSCs group compared to the control group. Histopathologic examination of the liver, following MSCs treatment, showed an apparently normal looking liver with no evidence of neoplastic cellular changes. The biochemical results support these findings. CONCLUSIONS: This work documents that MSCs could be labeled with nanoparticles and traced in normal and cirrhotic liver and in liver with HCC in animals using MRI. MRI monitors the homing and localization of MSCs in the liver. MSCs infusion in animal models of cirrhosis and carcinoma may prove to be useful in limiting the cirrhotic process. Also, it may have a possible therapeutic potential on the carcinogenic process.

Study of the effects of cyclooxygenase-2 inhibitor on the promotion of hepatic tumorigenesis in rats fed a high fat diet.

BACKGROUND/OBJECTIVE: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide. The highest prevalence of hepatitis is an important risk factor contributing to development of HCCs. However, an increasing number of cases are associated metabolic disease and steatohepatitis. Inflammation associated with many liver disease, seems to be a necessary pre-requisite for successful tumor initiation. Mechanisms that link high fat diet and inflammation initial stage of HCC are not completely understood. The present work was designed to investigate the effect of fat, through modulation of the insulin-like growth factors I and II (IGF-I and IGF-II), on the promotion of hepatocellular carcinoma, and the role of cyclooxygenase 2 (COX-2). METHODS: two main groups of rats were used: control and HCC groups. The HCC group was further sub-divide in to two subgroups, HCC fed with standard diet and HCC fed with high fat diet. The effects of celecoxib were also investigated in HCC fed with high fat diet. RESULTS: We found that high fat diet was associated with significant increases in COX2 and interleukin 6 (IL6) with significant promotion of HCC progression. The significant increase in IGF could contribute partially to the observed effects of high fat diet. In addition, celecoxib was found to significantly reduce HCC progression. CONCLUSIONS: We conclude that COX2 could play central role in high prevalence of HCC observed with high fat diet. Several triggering factors such as IGF and IL6, together with the direct modulation of fat metabolism could open several novel preventive strategies of celecoxib treatment, and could be useful biomarkers for assessment of its pharmacological effects.

The role of bone marrow derived-mesenchymal stem cells in attenuation of kidney function in rats with diabetic nephropathy.

BACKGROUND: Stem cell therapy holds a great promise for the repair of injured tissues and organs, including the kidney. We studied the effect of mesenchymal stem cells (MSC) on experimental diabetic nephropathy (DN) in rats and the possible paracrine signals that mediate their action. MATERIALS AND METHODS: Rats were divided into controls, DN rats, DN rats receiving MSCs. MSCs were given in a dose of (106cells) by intravenous injection. After 4 weeks, 24 h urinary albumin, serum urea and creatinine concentrations, transforming growth factor ß (TGF ß), tumor necrosis factor α (TNFα), B-cell lymphoma 2 (bcl2) and Bax gene expression and vascular endothelial growth factor (VEGF) were assessed. Histopathology staining was performed. RESULTS: MSC therapy significantly improved 24 h urinary albumin, serum urea and creatinine concentrations, increased angiogenic growth factor VEGF, and anti-apoptotic protein bcl2 while decreased the pro-inflammatory TNF-α, fibrogenic growth factor TGF ß, and pro-apoptotic protein Bax. The histopathology examination showed patchy areas of minimal necrosis and degeneration in renal tubules.

The effect of a novel curcumin derivative on pancreatic islet regeneration in experimental type-1 diabetes in rats (long term study).

BACKGROUND: Several studies highlight curcumin's benefit as a hypoglycemic agent, however; a limited number of reports present the importance of curcumin in improvement of pancreatic islets in diabetes. The aim of the present study is to evaluate the antidiabetic effect of a novel curcumin derivative and its effect on pancreatic islet regeneration in type I diabetes-induced by STZ. MATERIALS AND METHODS: Rats were divided into diabetic rats and diabetic rats treated orally with the novel curcumin derivative (NCD) for 40 days. Fasting blood samples were withdrawn periodically from all rats to estimate plasma glucose, insulin and C-peptide for 10 months. Histopathology was performed to allow the assessment of pancreatic islet morphology. Insulin and CD105 were detected immunohistochemically. RESULTS: In diabetic rats, the plasma glucose, insulin and C-peptide levels remained within the diabetic range for about 4 months, after which a gradual decrease in glucose and increase in insulin and C-peptide was observed, which reached almost normal levels after 10 months. NCD treated diabetic rats showed significantly lowered plasma glucose and increased plasma insulin and C-peptide levels. This was followed by a further significant decrease in plasma glucose and increase in plasma insulin and C-peptide after two months from oral administration of the NCD. The plasma insulin and C-peptide continued to increase for ten months reaching levels significantly higher than the basal level. Histopathological examination of diabetic rat pancreas revealed absence of islets of Langerhans, minimal adipose tissue infiltration and localized lymphocytic infiltrates. However, after 6 months of induction of diabetes, rat pancreas showed the appearance of small well formed islets and positive insulin cells but no CD105 positive cells. NCD treated rats showed the appearance of primitive cell collections, large insulin positive cells and CD105 positive cells in the adipose tissue infiltrating the pancreatic tissues. This was followed by the gradual appearance of insulin positive cells in the islets while, CD 105 positive cells remained in the adipose tissue. After 5 and 10 months from the onset of diabetes, rat pancreas showed, well developed larger sized islets with disappearance of primitive cell collections and CD 105 positive cells. Also, insulin positive islets of variable size with disappearance of insulin positive cells in adipose tissue were detected. CONCLUSION: The NCD possesses antidiabetic actions and enhanced pancreatic islets regeneration.

Image cytometry of fine needle aspiration of thyroid epithelial lesions.

Fine needle aspiration (FNA) biopsy has been documented as a reliable, cost-effective test, diagnosing different thyroid lesions. However, its sensitivity declines much in some settings. This study aimed to investigate the value of computerized morphometry and DNA ploidy in differential diagnosis of different thyroid epithelial follicular lesions diagnosed on FNA smears. In this study, 68 cases of thyroid FNA were cytologically examined, and classified according to their postthyroidectomy histopathologic diagnosis as follow: 10 cases of nodular goiter, 14 cases of adenomas, 10 cases of follicular carcinoma, 20 cases of papillary carcinoma, 10 cases of anaplastic carcinoma, and 4 cases of poorly differentiated carcinoma. Using Leica Qwin 500 image analysis system, nuclear area as a morphologic variable, and the DNA ploidy, DNA index, and proliferation indices were measured for each case. Sensitivity of FNA in the studied cases was optimum (100%) in papillary carcinomas and anaplastic carcinomas. However, the sensitivity declined sharply in discriminating nodular goiter and follicular adenoma from well-differentiated follicular carcinoma, thus, most of these cases were diagnosed as follicular lesions. A significant difference in mean values of nuclear area was found between benign and malignant lesions, with no statistical differences within the benign group. Aneuploidy was significantly associated with malignancy. Therefore, image analysis may become an important ancillary test in the initial evaluation of patients with thyroid epithelial lesions.

Changing patterns (age, incidence, and pathologic types) of schistosoma-associated bladder cancer in Egypt in the past decade.

OBJECTIVE: To assess the patterns of schistosomiasis-associated bladder cancer in Egypt from 2001 to 2010 in a retrospective study. Bilharzial bladder carcinoma is the most common cancer, particularly in Egyptian men. Classically, carcinoma in a bilharzial bladder is most commonly of the squamous cell type. During the past decade, certain changes have occurred in the features in Schistosomiasis-associated carcinoma in Egypt with a decline in the frequency of squamous cell carcinoma and increase in the frequency of transitional cell carcinoma. METHODS: This was a retrospective study of 1932 patients treated at Kasr Al Aini Hospital, Cairo University, from 2001 to 2010. Two groups were selected: group 1 included 1002 patients from 2001 to 2005 and group 2 included 930 patients from 2006 to 2010. RESULTS: The mean patient age increased from 41±11.2 years to 52±8.6 years, and the male/female ratio changed from 5.6:1 to 4.2:1. The incidence of associated bilharziasis decreased from 80% to 50%. A significant increased occurred in transitional cell carcinoma from 20% to 66%, with a significant decrease in squamous cell carcinoma from 73% to 25%. No difference was observed in the tumor stage or grade or incidence of lymph node metastases between the 2 groups. CONCLUSION: The pattern of incidence of the various histologic types of bladder cancer have changed, with most cases now transitional cell carcinoma, in contrast to the findings in the earlier Egyptian series. Additional studies are encouraged to explain the factors explaining these changes.

Efficacy of mesenchymal stem cells in suppression of hepatocarcinorigenesis in rats: possible role of Wnt signaling.

BACKGROUND: The present study was conducted to evaluate the tumor suppressive effects of bone marrow derived mesenchymal stem cells (MSCs) in an experimental hepatocellular carcinoma (HCC) model in rats and to investigate the possible role of Wnt signaling in hepato-carcinogenesis. METHODS: Ninety rats were included in the study and were divided equally into: Control group, rats which received MSCs only, rats which received MSCs vehicle only, HCC group induced by diethylnitroseamine (DENA) and CCl(4), rats which received MSCs after HCC induction, rats which received MSCs before HCC induction. Histopathological examination and gene expression of Wnt signaling target genes by real time, reverse transcription-polymerase chain reaction (RT-PCR) in rat liver tissue, in addition to serum levels of ALT, AST and alpha fetoprotein were performed in all groups. RESULTS: Histopathological examination of liver tissue from animals which received DENA-CCl(4) only, revealed the presence of anaplastic carcinoma cells and macro-regenerative nodules type II with foci of large and small cell dysplasia. Administration of MSCs into rats after induction of experimental HCC improved the histopathological picture which showed minimal liver cell damage, reversible changes, areas of cell drop out filled with stem cells. Gene expression in rat liver tissue demonstrated that MSCs downregulated ß-catenin, proliferating cell nuclear antigen (PCNA), cyclin D and survivin genes expression in liver tissues after HCC induction. Amelioration of the liver status after administration of MSCs has been inferred by the significant decrease of ALT, AST and Alpha fetoprotein serum levels. Administration of MSCs before HCC induction did not show any tumor suppressive or protective effect. CONCLUSIONS: Administration of MSCs in chemically induced HCC has tumor suppressive effects as evidenced by down regulation of Wnt signaling target genes concerned with antiapoptosis, mitogenesis, cell proliferation and cell cycle regulation, with subsequent amelioration of liver histopathological picture and liver function.

Immunohistochemical study of the expression of matrix metalloproteinase-9 in skin lesions of mycosis fungoides.

Matrix metalloproteinase-9 (MMP-9) has been correlated with poor clinical outcome in various malignancies and is associated with enhanced tumor growth and dissemination through its role in angiogenesis. This study was carried out to review the immunohistological staining of MMP-9 in skin lesions of different stages of mycosis fungoides (MF). The study was carried on 22 patients with MF and 10 healthy controls. Immunohistochemical staining using MMP-9 monoclonal anti-human antibodies was performed to determine the intensity of expression and distribution pattern of MMP-9 in MF lesions and in normal control skin. The general intensity of expression of MMP-9 was found to be significantly higher in cases with MF than in controls, and it increased in direct proportion to the increase in disease severity, being greatest in the tumor stages. A significantly greater number of blood vessels were found in cases with MF when compared with controls, and the MMP-9 expression by endothelial cells was significantly higher in endothelial cells within tumor cell aggregates than in endothelial cells outside the tumor cell aggregates. This study raises the possibility that MMP-9 may play an important role in the development of MF lesions, and its significantly higher expression in tumor stages may point to a possible role in disease progression. Further studies are needed to validate these findings and to assess the possible therapeutic role of drugs targeting MMP-9 in the treatment of MF.

An immunohistochemical study of laminin in basal cell carcinoma.

BACKGROUND: Laminins are components of the extracellular matrix that mediate cell adhesion, growth, migration, proliferation and differentiation. Basement membrane (BM) laminins, in particular, may play a role in enhancing carcinoma cell motility. AIM: To evaluate the distribution pattern of laminin in basal cell carcinoma (BCC), as regards the basement membrane, cellular cytoplasm, peritumoral lacunae and surface epithelium and to correlate laminin distribution with different variants of BCC. PATIENTS AND METHODS: Skin biopsy specimens were obtained from 21 BCC patients for routine histopathological and immunohistochemical study. Laminin was evaluated qualitatively and semiquantitatively using monoclonal mouse antihuman antibody (Dako-Laminin, 4C7. Code No: MO638, which reacts with the terminal globular domain of the α5 chain) RESULTS: The majority of BCC cases showed patchy cytoplasmic distribution of laminin. The BM expression of laminin, in most cases, was well defined, fine and linear with irregular areas of thickening. Staining intensity was moderate in differentiated and mixed variants, weak in superficial spreading and absent in morpheic types. CONCLUSION: Cytoplasmic and basement membrane laminin is important in the pathogenesis and invasion of BCC. Most laminin was in basement membrane zone (BMZ), and the more differentiated the tumor, the more cytoplasmic and BM staining it expressed.

Effects of losartan, HO-1 inducers or HO-1 inhibitors on erectile signaling in diabetic rats.

INTRODUCTION: Activation of the renin-angiotensin system which is common in diabetes mellitus might affect heme oxygenase (HO-1) gene expression. AIM: Assessment of the effects of administration of angiotensin II (Ang II) receptor antagonist (losartan) with HO-1 inducer or inhibitor on erectile signaling in diabetic rats. MATERIALS AND METHODS: Seventy male rats were divided equally into seven groups; healthy controls, streptozotocin-induced diabetic rats, rats on citrate buffer, diabetic rats on losartan, diabetic rats on HO-1 inducer (cobalt protoporphyrin [CoPP]), diabetic rats on losartan and CoPP, and diabetic rats on losartan and HO-1 inhibitor (stannus mesoporphyrin [SnMP]). MAIN OUTCOME MEASURE: HO enzyme activity, HO-1 gene expression, cyclic guanosine monophosphate (cGMP) assay, intracavernosal pressure (ICP), and cavernous tissue sinusoids surface area. RESULTS: HO-1 gene expression, HO enzymatic activity, and cGMP were significantly decreased in the cavernous tissue of diabetic rats. These parameters were significantly elevated with the use of CoPP that restored the normal control levels of HO enzyme activity. Administration of losartan exhibited a significant enhancing effect on these parameters compared with the diabetic group, but not restored to the control levels, whereas administration of CoPP combined with losartan led to the restoration of their normal levels. ICP demonstrated significant decline in diabetic rats. The use of CoPP and/or losartan led to its significant improvement compared with diabetic rats. Administration of either losartan and/or CoPP led to a significant increase in the cavernous sinusoids surface area of diabetic rats. Administration of losartan with SnMP significantly decreased the enhancing effect of losartan on the studied parameters. CONCLUSION: The decline in erectile function in diabetes mellitus could be attributed to the downregulation of HO-1 gene expression. HO-1 induction added to Ang II receptor antagonist could improve erectile function.

Immunohistochemical study of CD99 and EMA expression in ependymomas.

Tumors of the central nervous system (CNS) represent a unique, heterogeneous population of neoplasms and include both benign and malignant tumors. The present study was carried out on a total of 79 archival cases of ependymal tumors in addition to a variety of other primary CNS tumors. The study entailed the use of CD99 monoclonal antibody and epithelial membrane antigen (EMA). It was found that all 38 ependymoma cases (classic and nonclassic) showed positive membranous and intracytoplasmic CD99 immunoreactivity. Upon comparing with other CNS tumors (41 cases), it was found that CD99 could differentiate between ependymomas and nonependymal tumors, but intensity and pattern of staining were of no consequence in determining variant type or degree of histologic aggressiveness. In regard to EMA immunoreactivity, which was restricted to the ependymoma group, 2 patterns of staining could be detected--the intracytoplasmic dotlike pattern and the ringlike pattern--but some cases were completely negative. Thus, EMA was found to be of little value in the diagnosis of ependymoma and in the differentiation between different types and grades. CD99 can hence be recommended for use as a good marker for differentiation between ependymal and other CNS tumors. EMA expression and pattern of distribution, on the other hand, cannot be employed to determine the type of variant or the degree of tumor aggressiveness, and hence cannot predict the behavior of ependymal neoplasms.

Heme oxygenase vs. nitric oxide synthase in signaling mediating sildenafil citrate action.

INTRODUCTION: Heme oxygenase (HO) enzyme catalyzes the rate limiting step in oxidative degradation of heme to biliverdin and carbon monoxide (CO). CO has been shown to share many properties with nitric oxide (NO), including activation of guanyl cyclase, signal transduction, and gene regulation. AIM: To assess the signaling pathways mediating cavernous tissues response to sildenafil citrate intake experimentally. MAIN OUTCOME MEASURES: In dissected cavernous tissues; detection of HO-1, HO-2 and nueronal nitric oxide synthase (nNOS) gene expressions by reverse transcriptase polymerase chain reaction (RT-PCR), HO enzyme activity assay, HO-1, HO-2 protein detection by Western blot, cyclic guanosine monophosphate (cGMP) tissue levels by enzyme linked immunosorbent assay (ELISA) and histopathology. METHODS: Two hundred forty Sprague-Dawley rats divided into five equal groups were investigated: group (Gr) 1, controls received regular diet; Gr 2, received sildenafil citrate 4 mg/kg orally; Gr 3, received the same dose of sildenafil added to HO inducer, diferuloylmethane; Gr 4, received sildenafil added to HO inhibitor, zinc protoporphyrin, and Gr 5, received sildenafil kg orally by gastric tube. Gr 3 received the same dose of sildenafil added to HO inducer, added to nitric oxide synthase inhibitor, L-Nitroarginine methylester. Twelve rats from each group were sacrificed by cervical dislocation successively after 1/2, 1, 2, and 3 hours from the intake. RESULTS: HO-2 gene expression was demonstrated in all groups. HO-1 was not expressed in controls, expressed in Gr 2, accentuated in Gr 3, and attenuated in Gr 4 and 5. These results were confirmed by Western blot. The nNOS was expressed in controls, increased in Gr 2 and 3, and decreased in Gr 4 and 5. HO enzyme activity and cGMP levels were significantly elevated in Gr 2, accentuated in Gr 3, and significantly decreased in Gr 4 and 5 compared to controls. Vasodilatations were observed in cavernous tissues of histopathologic sections of Gr 2 and increased in those of Gr 3. CONCLUSION: Sildenafil citrate actions may be mediated by up-regulation of HO-1 gene expression.