Fadia M. Attia

Novel role of curcumin combined with bone marrow transplantation in reversing experimental diabetes: Effects on pancreatic islet regeneration, oxidative stress, and inflammatory cytokines.

Therapeutic utility of bone marrow transplantation in diabetes is an attractive approach. However, the oxidative stress generated by hyperglycemia may hinder β-cell regeneration. The present study was undertaken to investigate the therapeutic potential of curcumin, a dietary spice with antioxidant activity, bone marrow transplantation, and their combined effects in the reversal of experimental diabetes. Diabetes was induced in mice by multiple low doses of streptozotocin. After the onset of diabetes, mice were treated with curcumin (10 mM; 100 μl/mouse, i.p., for 28 days) or received a single bone marrow transplantation (10(6) un-fractionated bone marrow cells), or both. Parameters of diabetes, integrity of pancreatic islets, pancreatic oxidative stress markers, and serum pro-inflammatory cytokines, were evaluated. Treatment with either curcumin or bone marrow transplantation significantly reversed streptozotocin-induced hyperglycemia/glucose intolerance, hypoinsulinemia, and damage of pancreatic islets. Interestingly, combination of curcumin and bone marrow transplantation elicited the most profound alleviation of such streptozotocin-evoked anomalies; including islet regeneration/insulin secretion. On the other hand, curcumin, either alone or combined with bone marrow transplantation, blunted the pancreatic lipid-peroxidation, up-regulated activities of the antioxidant enzymes, and suppressed serum levels of TNF-α and IL-1β. Curcumin and single bone marrow transplantation proved their therapeutic potential in reversing diabetes when used in combination. Curcumin, via its antioxidant and anti-inflammatory effects, evidently enhanced the ability of bone marrow transplantation to regenerate functional pancreatic islets. Hence, the use of natural antioxidants combined with other therapeutic regimens to induce pancreatic regeneration is a promising strategy in the management of diabetes.

Evaluation of neutrophilic CD64, interleukin 10 and procalcitonin as diagnostic markers of early- and late-onset neonatal sepsis.

Abstract The assay of infection markers can improve diagnostic sensitivity in neonatal sepsis. We determined the levels of neutrophilic CD64 (nCD64), procalcitonin (PCT) and interleukin 10 (IL-10) in infants with neonatal sepsis. Forty-nine newborn infants who met the criteria of sepsis were subjected to a routine sepsis evaluation as well as measurement of PCT and IL-10 levels and nCD64 expression. Of these 49 ‘infected’ infants, 16 had a positive blood culture (culture-positive sepsis) and 33 infants were diagnosed to have clinical sepsis with negative blood cultures (culture-negative sepsis). Another 49 healthy newborn infants were included as a control group. The sensitivity, specificity, positive predictive value and negative predictive value of PCT, IL-10 and nCD64 for the diagnosis of sepsis were determined. IL-10 had the highest sensitivity of 92% and specificity of 84% using a cut-off of ≥17.3 pg/ml. For PCT, the highest sensitivity of 65% and specificity of 60% were found at a cut-off value of ≥36.4 pg/ml. nCD64 had a maximal sensitivity of 92% and specificity of 71% at a cut-off value of 2.6%. Combinations of different markers may improve the sensitivity and specificity of biomarker tests. We found that the best combination was IL-10 and nCD64, which together provided sensitivity of 95% and specificity of 83%, and a negative predictive value of 86%.

Multiple intra-familial transmission patterns of hepatitis B virus genotype D in north-eastern Egypt†

The transmission rate of intra‐familial hepatitis B virus (HBV) and mode of transmission were investigated in north eastern Egypt. HBV infection was investigated serologically and confirmed by molecular evolutionary analysis in family members (N = 230) of 55 chronic hepatitis B carriers (index cases). Hepatitis B surface antigen (HBsAg) and hepatitis B core antibody (anti‐HBc) prevalence was 12.2% and 23% among family members, respectively. HBsAg carriers were prevalent in the age groups; <10 (16.2%) and 21–30 years (23.3%). The prevalence of HBsAg was significantly higher in the family members of females (19.2%) than males (8.6%) index cases (P = 0.031). HBsAg and anti‐HBc seropositive rates were higher significantly in the offspring of females (23%, 29.8%) than those of the males index cases (4.3%, 9.8%) (P = 0.001, 0.003), as well as higher in the offspring of an infected mother (26.5, 31.8%) than those of an infected father (4.7%, 10.5%) (P = 0.0006, 0.009). No significant difference was found in HBsAg seropositive rates between vaccinated (10.6%) and unvaccinated family members (14.8%). Phylogenetic analysis of the preS2 and S regions of HBV genome showed that the HBV isolates were of subgenotype D1 in nine index cases and 14 family members. HBV familial transmission was confirmed in five of six families with three transmission patterns; maternal, paternal, and sexual. It is concluded that multiple intra‐familial transmission routes of HBV genotype D were determined; including maternal, paternal and horizontal. Universal HBV vaccination should be modified by including the first dose at birth with (HBIG) administration to the newborn of mothers infected with HBV. J. Med. Virol. 84:587–595, 2012.

Circulating Endothelial Cells as a Marker of Vascular Dysfunction in Patients With Systemic Lupus Erythematosus by Real-Time Polymerase Chain Reaction

CONTEXT Systemic lupus erythematosus (SLE) is associated with an increased risk of atherosclerosis; endothelial dysfunction represents the first step in its pathogenesis. OBJECTIVE To assess endothelial dysfunction in SLE by circulating endothelial cells (CECs) and to characterize SLE-specific factors that contribute to its appearance. DESIGN Case-control study was conducted on 60 subjects, divided into 2 groups: group A (30 patients with SLE) and group B (30 healthy sex- and age-matched controls). Total cholesterol, triglycerides, antinuclear antibodies, anti-double-stranded DNA antibodies, and C3 were determined in all patients. Systemic lupus erythematosus activity was assessed using the SLE Disease Activity Index. Endothelial function was assessed by means of flow-mediated dilation of the brachial artery using B-mode ultrasonography and relative quantification of CD 146 mRNA by real-time polymerase chain reaction. RESULTS The group of SLE patients was formed of 20 females and 10 males, with a mean age of 31.16 ± 9.69 years. The values of SLE-specific tests and SLE Disease Activity Index were represented by anti-double-stranded DNA antibodies 160 ± 40.5, C3 68.91 ± 11.91 mg/dL, total cholesterol 188.66 ± 49.63 mg/dL, triglycerides 143.41 ± 46.26 mg/dL, and SLE Disease Activity Index 12.66 ± 3.70. Values for flow-mediated dilation were 8.85% ± 2.02% (group A) and 20.33% ± 6.19% (group B), P < .001, and CECs were 300 ± 40.5 μL⁻¹ blood (group A) and 10 ± 2.5 μL⁻¹ blood (group B). The statistical analysis showed a strong inverse correlation between CECs and SLE Disease Activity Index, a strong correlation between CECs and C3, a strong correlation between CECs and anti-double-stranded DNA antibodies, and a moderate inverse correlation between CECs and total cholesterol. CONCLUSION Endothelial dysfunction is present in SLE patients even in the absence of traditional cardiovascular risk factors due to disease activity.

Thrombocytopenia in Patients With Chronic Hepatitis C: A Possible Role of HCV on Platelet Progenitor Cell Maturation

A total of 30 patients with chronic hepatitis C (HCV) thrombocytopenia (TP) and 20 healthy controls were studied. Both groups were subjected to complete medical history, clinical examination in addition to assessment of hepatitis markers: level of thrombopoietin (Tpo), Geimsa-stained bone marrow smears, and in vitro short-term megakaryocytic progenitors culture (CFU-MK). Serum Tpo level was significantly elevated in patients with TP HCV. Short-term CFU-MK showed an evident depression in the colony-forming unit—megakaryocyte (CFU-meg). There is a positive correlation between the number of CFU-meg and the platelet count and between serum Tpo level and prothrombin time, transaminase, albumin, and the Child Pugh score of liver disease; a negative correlation between serum Tpo level and the number of CFU-meg and between serum Tpo level and the platelet count. Thus, the level of Tpo could be an indicator of intact functional response of the hepatocytes.

Clinical significance of suppressor of cytokines signalling-3 mRNA expression from patients with non-Hodgkin lymphoma under chemotherapy.

INTRODUCTION To date, little is known about blood immune marker changes that may be related to the development of Non Hodgkin Lymphoma (NHL) and treatment response with few serum biomarkers that could be useful in follow- up of the patients. OBJECTIVE To quantify the expression of suppressor of cytokine signalling-3-(SOCS-3) gene at the mRNA level in the peripheral blood of patients with NHL and correlate with clinical pathological features and response to treatment. METHODS Thirty patients with NHL and 20 healthy controls were enrolled in the study. The SOCS-3 mRNA level in peripheral blood (PB) was detected by semi-quantitative real-time polymerase chain reaction. Quantification of cytokines such as interleukin 6 and tumour necrosis factor alpha (IL-6 & TNF-α) were performed using sandwich enzyme-linked immunosorbent assays (ELISA). RESULTS Increased expression of SOCS-3 mRNA in peripheral blood plus increased serum levels of IL-6 and TNF alpha from NHL cases with no complete remission after therapy. Higher levels of expression of SOCS-3 are associated with advanced disease, bone marrow involvement, extranodal involvement, poor performance status, B cell symptoms (fever, night sweats and weight loss) and high serum lactate dehydrogenase level which are evaluated by international prognostic index (IPI). Complete responses occur in 60% of patients with normal expression of SOCS-3 gene. Increased expression of SOCS-3 is common in diffuse large B cell lymphoma, CLL/small lymphocytic B cell lymphoma and follicular lymphoma. CONCLUSIONS Over-expression of SOCS-3 mRNA from peripheral blood of NHL patients correlates with advanced disease and poor response to treatment. SOCS-3 mRNA expression in peripheral blood from NHL patients might be used to monitor response during treatment.

The prognostic value of p53 mutation in pediatric marrow hypoplasia

BackgroundThe tumor suppressor gene p53 is involved in the control of cell proliferation, particularly in stressed cells. p 53 gene mutations are the most frequent genetic event found in human cancers. Fanconi Anemia (FA) is the most common representative of inherited bone marrow failure syndromes (IBMFS) with a leukemic propensity. P 53 DNA alteration has not been studied before in Egyptian children with FA.Patients and methodswe investigated p53 mutation in the bone marrow and peripheral blood of forty children, FA (n = 10), acquired aplastic anemia (AAA) (n = 10), and immune thrombocytopenia (ITP) as a control (n = 20), using real-time PCR by TaqMan probe assayResultsMutation of p53 gene was demonstrated in the BM of 90% (9/10) of children with FA, compared to 10% (1/10) in AAA (p < 0.001), while, no p53 DNA mutation was seen in the control group. A positive correlation between DNA breakage and presence of p53 mutation was seen in FA (p < 0.02, r0.81).Conclusionmutation of p53 gene in hypoplastic marrow especially FA may represent an early indicator of significant DNA genetic alteration with cancer propensity.

Effect of Schwann and mesenchymal stem cells on experimentally induced sciatic nerve injury in rats

Objective Transplantation of bone marrow stromal cells (BMSCs) and Schwann cells (SCs) can facilitate axon regeneration in peripheral nerve injuries. The aim of this study was to explore the effect of transplantation of BMSCs and SCs on electrophysiological recording after injury of the sciatic nerve in the rat. Materials and methods In this study, 40 adult male albino rats (250-300 g) were used. BMSCs and SCs were cultured. Rats were divided randomly into four equal groups: group 1, control without nerve injury; group 2, nerve injury without cell transplantation; group 3, nerve injury with BMSCs transplantation; and group 4, nerve injury with SCs transplantation. Standardized crush injury of the sciatic nerve (axonotmesis) was performed by surgical hemostat at the first lock for 1 min; BMSCs and SCs were separately transplanted intralesionally. After 8 weeks, electrophysiological recordings were analyzed by one-way analysis of variance. Results Electrophysiological analysis showed a significant improvement in the cell transplantation groups compared with the injured group (P < 0.01). Conclusion BMSCs and SCs may potentially enable recovery after a standardized injury to the sciatic nerve in rats (axonotmesis). Electrophysiological evaluation confirms this improvement after transplantation of SCs and BMSCs, with no significant difference between them.

Prognostic value of VEGF mRNA expression in short-term follow-up of patients with acute coronary syndrome

Acute coronary syndrome (ACS) has a complex and heterogeneous pathophysiology, with patients being at high risk of death and cardiac ischemic events. This study aimed to evaluate the prognostic value of VEGF gene expression in patients with acute coronary syndrome and to correlate gene expression of VEGF with the short-term outcome of ACS. This descriptive study included 28 patients with acute coronary syndrome from Suez Canal University Hospital. Diagnosis of patients with acute coronary syndrome was based on history taking, clinical examination, resting ECG, and cardiac enzymes. VEGF mRNA expression was assessed by Taqman technique using real time PCR method. Results showed that there was significantly higher mean VEGF mRNA in the patients than in the control subjects (p < 0.05). Regarding the relation between good and bad outcomes versus low and high VEGF of the studied patients, there was no statistically significant difference between VEGF in the patients with bad and good outcomes (p > 0.05). VEGF mRNA has a diagnostic role in ACS VEGF. It may play a role in the prognosis of ACS after long-term period but not in a short-term period as in our study.

Flowcytometric analysis of aldehyde dehydrogenase activity in mononuclear cells from umbilical cord blood

Aldehyde dehydrogenase (ALDH) is a cytosolic enzyme that is responsible for the oxidation of intracellular aldehydes. Elevated levels of ALDH have been demonstrated in murine and human progenitor cells compared with other hematopoietic cells, and this is thought to be important in chemoresistance and purification techniques and an indication of the proper function of the cell. A Flowcytometric method for the assessment of ALDH activity in viable cells recently has been developed. Forty six cord blood samples from mothers which underwent normal delivery of full term infants were obtained, after informed consent. Mononuclear cells were obtained by Ficoll-Paque density centrifugation and ammonium chloride red cell lysis. Percentage of viable cells was determined by trypan blue exclusion dye. Cells were labeled with Aldefluor reagent (Vancouver Canada) as described by the manufacturer. Cells were then stained with phycoerythrin (PE)–conjugated anti-CD34 (Miltenyi Biotec, Cologne, Germany) antibodies for 30 min at 4°C. Cells were washed and re-suspended in phosphate-buffered saline (PBS) with 2% fetal calf serum. Cells were then analyzed on coulter epics flow cytometer. The mean percentage of ALDH enzyme expression among the CD34+ cells in the cord blood samples was 61.3% with a minimum of 28% and a maximum of 94.6%. Significant correlations were found between the white blood cell (WBCS) count in the cord blood samples and both the CD34+ cell count and the count of ALDH expressing cells, while no correlation was found between the CD34+ cells count or the ALDH expressing cells count in the cord blood samples and either the sex or the weight of the newborn. Identification and isolation of cells on the basis of ALDH activity provides a tool for their isolation and further analysis. In summary, a high ALDH-1 activity identifies CD34+ cells in cord blood. Key words: Umbilical cord blood, stem cells, aldehyde dehydrogenase (ALDH), CD34.