The ameliorating effects of mesenchymal stem cells compared to α-tocopherol on apoptosis and autophagy in streptozotocin-induced diabetic rats: Implication of PI3K/Akt signaling pathway and entero-insular axis.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) are considered a novel regenerative therapy that holds much potential. This study aimed to examine and compare the ameliorative effects of BM-MSCs compared to α-tocopherol (α-Toc) on apoptosis, autophagy, and ß-cell function in a rat model of streptozotocin (STZ)-induced diabetes and further analyzed the implications and interrelations of the entero-insular axis, and type I phosphoinositide 3-kinase (PI3K)/Akt signaling. Forty adult male albino rats were categorized into four groups (n = 10, in each): control group, STZ-induced diabetic group (single i.p. injection of STZ 45 mg/kg), diabetic and treated with BM-MSCs injection, diabetic and treatment with α-Toc p.o. The serum glucose, insulin, nitric oxide (NO), and catalase (CAT) were measured. Histopathological examination of the pancreas, the expression levels of insulin, CD44, caspase-3, autophagy markers, P13K/Akt, and pancreas/duodenum homeobox protein 1, in pancreatic tissue, and glucose-dependent insulinotropic polypeptide (GIP) in the duodenum were detected by hematoxylin and eosin staining, immunofluorescence labeling, and by quantitative real-time polymerase chain reaction. The diabetic rats showed reduced insulin, hyperglycemia, nitrosative stress (NO, CAT), augmented apoptosis (caspase 3), impaired autophagy (p62/SQSTM1, LC3), downregulated PI3K/Akt pathway and increased GIP expression, and degeneration of pancreatic islets. Treatment with either BM-MSCs or α-Toc suppressed the nitrosative stress, reduced apoptosis, recovered autophagy, upregulated PI3K/Akt pathway, and subsequently increased insulin levels, decreased blood glucose, and downregulated GIP expression with partial restoration of pancreatic islets. Based on our findings, the cytoprotective effects of BM-MSCs and α-Toc in type 1-induced diabetes appeared to be related to repaired autophagy and recovered PI3K/Akt signaling. Moreover, we reported their novel effects on reversing intestinal GIP expression level. The effect of BM-MSCs was notably superior to that of α-Toc.

Evaluation of trail receptor 1 (DR4) polymorphisms C626G and A683C as risk factors of hepatocellular carcinoma.

Tumor necrosis factor related apoptosis-inducing ligand (TRAIL) plays an important role in many cancers including hepatocellular carcinoma (HCC). The aim of this study is to investigate the association of the DR4 polymorphisms C626G (Thr209Arg, rs20575) and A683C (Glu228Ala, rs20576) with the occurrence of HCC in Egyptian patients chronically infected with HCV. The study included 80 patients with HCV-related HCC (group 1) and 80 patients with HCV-related liver cirrhosis (group 2) who are naïve to treatment. Clinical and laboratory data were recorded. Genotyping of TRAIL receptor DR4 polymorphism C626G rs20575 and A683C rs20576 SNP was done by Real-Time PCR using taqman probes technology. The mean age of HCC patients was 57.6 ± 8.4 years with 62 patients (77.5%) were males. While group 2 mean age was 49.5 ± 10.29 years with 50% were males. The frequency distribution of rs20575 genotypes showed a statistically significant difference between the two studied groups (P = 0.02), the carriers of the C allele were 2.01 times more likely to develop HCC than the carriers of the G allele (P = 0.003), while no significant difference in rs20576 genotypes distribution was found between the studied groups (P = 0.680). On combining the carriers of C allele of rs20575 and the carriers of A allele of rs20576, a significant difference was detected (P > 0.001) with 2.85 higher risk of HCC development in patients who carried both genetic risk alleles simultaneously. The significant difference in DR4 polymorphisms among HCC and cirrhotic patients suggests their role as potential risk factors of HCC development.

Transdermal Ketoprofen Patch in Comparison to Eutectic Mixture of Local Anesthetic Cream and Subcutaneous Lidocaine to Control Pain Due to Venous Cannulation.

BACKGROUND: This study was established to compare the analgesic and side effects between transdermal ketoprofen patch 30 mg and eutectic mixture of local anesthetic (EMLA) cream applied to the peripheral venous cannulation site and lidocaine injection before cannulation. PATIENTS AND METHODS: One hundred and five adult patients who had been scheduled for elective general surgery with patients' physical status American Society of Anesthesiologists classes I and II were randomly divided into three groups: Group I (EMLA group) received EMLA cream, Group II (lidocaine group) received subcutaneous infiltration of 1 ml of 2% lidocaine HCl 10 min before cannulation, and Group III (ketoprofen group) received a transdermal ketoprofen patch 30 mg. Groups I and III received their cream or patch 60 min before cannulation. The pain resulting from cannulation by an 18G cannula was assessed by a visual analog scale (VAS) at the time of cannulation and every 2 h for another 6 h for all groups. Signs of inflammation at the site of cannulation (erythema, induration, edema, and blanching) were observed at the site of cannulation for 24 h. RESULTS: Ketoprofen patch, EMLA cream, and lidocaine injection were found to be equal in controlling pain caused by venous cannulation with no significant difference in VAS. Signs of inflammation at the site of cannulation (blanching, erythema, and induration) were very evident in Group I (EMLA) which showed significant difference than in other two groups. CONCLUSIONS: EMLA cream, ketoprofen patch, and lidocaine injection have equal ability to alleviate pain due to cannulation when applied before the procedure, but ketoprofen patch is more superior as it had less local inflammatory effect in comparison to EMLA cream and without double puncture as with lidocaine injection.

Matrix Metalloproteinase-9 and Recovery of Acute Ischemic Stroke.

BACKGROUND: Stroke outcome can be predicted by clinical features, biochemical parameters, and some risk factors. Matrix metalloproteinase-9 (MMP-9) is involved in various stages of stroke pathology. MMP-9 inhibitors are potential stroke therapeutic agents. Little is known about the relation between MMP-9-after the acute stage-and clinical recovery. OBJECTIVE: The study aimed to investigate the serum level of MMP-9 at stroke onset as predictor of stroke outcome and the relation between the level of MMP-9 after 30 days and stroke recovery. METHODS: The National Institutes of Health Stroke Scale, modified Rankin Scale, and serum level of MMP-9 were assessed in 30 patients with acute ischemic stroke during the first 24 hours of onset and then a month later. None of the patients received thrombolytic therapy. Thirty normal volunteers of matched age and sex were included in the control group. RESULTS: The serum level of MMP-9 at stroke onset was independently positively correlated with stroke outcome. The serum level of MMP-9 30 days after stroke onset was positively correlated with initial stroke severity and outcome, as well as with clinical recovery. CONCLUSION: Higher serum level of MMP-9 at stroke onset can be a predictor of poor stroke outcome. However, beyond the acute stage, MMP-9 may play beneficial role in stroke recovery.

Relationship between matrix metalloproteinase-9 and common carotid artery intima media thickness.

Atherosclerosis causes significant morbidity and mortality. Carotid intima media thickness (IMT) predicts future ischaemic strok e incidence. Matrix metalloproteinases (MMPs) play a considerable role in atherosclerosis and hold therapeutic promise as well. To investigate the relationship between serum level of matrix metalloproteinase-9 (MMP-9) and common carotid artery intima media thickness (CCA-IMT) in patients with ischaemic stroke and asymptomatic subjects. Thirty patients with a previous ischaemic stroke and 30 asymptomatic volunteers were recruited. Assessment of vascular risk factors, serum level of MMP-9 and CCA-IMT on both sides was performed. The IMT of both CCAs correlated positively with the serum MMP-9 level in asymptomatic subjects (p = 0.000), even after adjustment for other risk factors. In the patients group, this positive correlation was significant for the right but not for the left CCA (right CCA: p = 0.023, left CCA: p = 0.0284). Fasting blood sugar correlated positively with serum levels of MMP-9 in asymptomatic subjects (p = 0.005) but did not correlate positively in patients. There was no significant correlation between MMP-9 and age or other investigated laboratory risk factors in either the patient or asymptomatic groups. MMP-9 is positively correlated with CCA-IMT both in stroke patients and asymptomatic subjects. This may indicate that MMP-9 is a possible therapeutic target for stroke prevention.

Triple test screening for Down syndrome: an Egyptian-tailored study.

BACKGROUND: The incidence of Down syndrome (DS) in Egypt varies between 1:555 and 1:770 and its screening by triple test is becoming increasingly popular nowadays. Results, however, seem inaccurate due to the lack of Egyptian-specific information needed for risk calculation and a clear policy for programme implementation. Our study aimed at calculation and validation of the triple marker medians used in screening Egyptian females as well as to recommend programme conventions to unify screening in this country. METHODS: The study was conducted on 668 Egyptian women, in weeks 15-20 of pregnancy as proven by sonar. Chorionic gonadotropin (CG), α-fetoprotein (AFP) and unconjugated oestriol (uE3) were measured on Siemens Immulite analyzer. Medians of the three parameters were calculated, regressed against gestational age (GA) and weighted by the number of participants/week. Equations were derived to adjust each parameter to the maternal weight and were centered on the median Egyptian weight. Prisca software was fed with the above data, multiples-of-median (MoM) and DS risks were calculated and the screening performance was evaluated at a mid-trimester risk cutoff of 1:270. RESULTS: Log-linear [AFP/uE3 = 10(A+B*GA)] and exponential equations [CG = A*e (B*GA)] were derived and the regressed medians were found to follow similar patterns to other Asian and Western medians. Oestriol was always lowest (even halved) while CG and AFP were intermediate. A linear reciprocal model best fitted weight distribution among Egyptians and successfully adjusted each parameter to a weight of 78.2 kg. Epidemiological monitoring of these recommendations revealed satisfactory performance in terms of 6.7% initial positive rate and 1.00 grand MoM. CONCLUSIONS: Adoption of the above recommendations is hoped to pave the way to a successful DS screening programme tailored to Egyptian peculiarities.

Pregnancy outcomes of expectant management of stable mild to moderate chronic hypertension as compared with planned delivery.

OBJECTIVE: To compare outcomes between elective delivery at 37 weeks of pregnancy and expectant management among pregnant women with mild to moderate chronic hypertension. METHODS: In a two-center study, 76 women with mild to moderate chronic hypertension were randomly allocated to planned delivery at 37 completed weeks (group A) or expectant management for spontaneous onset of labor or reaching 41 weeks (group B) between April 2012 and October 2013. Differences were compared by t test, χ(2) test, or Fisher exact test. Odds ratios (ORs) with 95% confidence interval (CIs) were determined. RESULTS: There were no differences in superimposed pre-eclampsia (SPE), severe hypertension, preterm delivery, placental abruption, oligohydramnios, intrauterine growth restriction, or perinatal mortality between the groups. Group B had higher gestational age at delivery (P=0.001) and birth weight (P=0.01), but lower cesarean (OR 3.4; 95% CI, 1.2-10.3; P=0.03) and neonatal care unit admission (OR 5.4; 95% CI, 1.4-21.0; P=0.01) rates. More women with SPE were diagnosed before than after 37 weeks in group B (P=0.01). Overall, patients who developed SPE had more adverse pregnancy outcomes than those who did not. CONCLUSION: Mild to moderate chronic hypertension could be managed expectantly up to 41 weeks if SPE did not develop.

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.

Effect of exercise training and anabolic androgenic steroids on hemodynamics, glycogen content, angiogenesis and apoptosis of cardiac muscle in adult male rats.

OBJECTIVES: To investigate the effects of exercise training and anabolic androgenic steroids (AAS) on hemodynamics, glycogen content, angiogenesis, apoptosis and histology of cardiac muscle. METHODS: Forty rats were divided into 4 groups; control, steroid, exercise-trained and exercise-trained plus steroid groups. The exercise-trained and trained plus steroid groups, after one week of water adaptation, were exercised by jumping into water for 5 weeks. The steroid and trained plus steroid groups received nandrolone decanoate, for 5 weeks. Systolic blood pressure and heart rate (HR) were monitored weekly. Heart weight/body weight ratio (HW/BW ratio) were determined. Serum testosterone, vascular endothelial growth factor (VEGF), cardiac caspase-3 activity and glycogen content were measured. RESULTS: Compared with control, the steroid group had significantly higher blood pressure, HR, sympathetic nerve activity, testosterone level, HW/BW and cardiac caspase-3 activity. Histological examination revealed apoptotic changes and hypertrophy of cardiomyocytes. In exercise-trained group, cardiac glycogen, VEGF and testosterone levels were significantly higher while HR was significantly lower than control. HW/BW was more than control confirmed by hypertrophy of cardiomyocytes with angiogenesis on histological examination. Trained plus steroid group, had no change in HR, with higher blood pressure and HW/BW than control, cardiac glycogen and serum VEGF were higher than control but lower than exercise-trained group. Histological examination showed hypertrophy of cardiomyoctes with mild angiogenesis rather than apoptosis. CONCLUSION: When exercise is augmented with AAS, exercise-associated cardiac benefits may not be fully gained with potential cardiac risk from AAS if used alone or combined with exercise.

Does cutaneous lupus erythematosus have more favorable pregnancy outcomes than systemic disease? A two-center study.

OBJECTIVE: To compare pregnancy outcomes in cutaneous lupus erythematosus (CLE) with systemic lupus erythematosus (SLE) and healthy pregnant women. DESIGN: Cohort comparative study. SETTING: Two university maternity centers in Saudi Arabia and Egypt. POPULATION: Pregnant women with CLE and SLE and healthy pregnant women. METHODS: Over a three-year period, 201 participants were allocated to three groups: group 1 (n = 67) contained women with CLE, group 2 (n = 67) women with SLE, and group 3 healthy controls (n = 67). Diagnosis of lupus erythematosus was based on American College of Rheumatology criteria. All participants were followed until delivery. Lupus exacerbation was evaluated by Lupus Activity Index score. ANOVA and chi-squared tests were used to compare obstetrical and neonatal outcomes, and regression analysis was used to define independent factors of adverse pregnancy outcomes. MAIN OUTCOME MEASURES: Pregnancy losses, preterm labor, intrauterine growth restriction, preeclampsia, neonatal intensive care unit admissions, cesarean sections and lupus exacerbations. RESULTS: There was no significant difference between groups 1 and 3 in rates of pregnancy loss, preterm labor, preeclampsia, intrauterine growth restriction and neonatal intensive care admission. Group 1 had lower pregnancy loss (p = 0.005), growth restriction (p = 0.001), preeclampsia (p = 0.05), neonatal intensive care admissions (p = 0.001), cesarean section (p = 0.03), lupus exacerbations (p = 0.05) and anti-phospholipid antibodies (p = 0.02) compared with group 2. In groups 1 and 2, lupus exacerbation and anti-phospholipid antibodies were significant independent factors for adverse outcomes. CONCLUSIONS: Cutaneous lupus erythematosus means comparable pregnancy outcomes to those of the healthy population. Lower rates of disease exacerbation and anti-phospholipid antibodies are potential factors for better pregnancy outcome in CLE compared with SLE.

Association of folate intake, dietary habits, smoking and COX-2 promotor -765G>C polymorphism with K-ras mutation in patients with colorectal cancer.

BACKGROUND: Understanding the role of environmental and molecular influences on the nature and rate of K-ras mutations in colorectal neoplasms is crucial. COX-2 polymorphisms -765G>C may play a role in carcinogenic processes in combination with specific life-style conditions or dependent on the racial composition of a particular population. If mutational events play an important role in colorectal carcinogenesis sequence, one can hypothesize that modification of these events by life-style or other factors would be a useful prevention strategy. AIM OF WORK: To explore the association between K-ras mutation and potential variables known or suspected to be related to the risk of colorectal cancer (CRC) as well as determining the possible modulating effect of the COX-2 polymorphism, -765G>C. SUBJECTS AND METHODS: The study was conducted on 80 patients with colorectal cancer from Tropical Medicine and Gastrointestinal Tract endoscopy Departments and those attending clinic of the National Cancer Institute, Cairo University during the period extending from April 2009 to March 2010. Full history taking with emphasis on the risk factors of interest, namely age, sex, family history, smoking and dietary history. Serum CEA and CA19-9, RBCs folic acid and occult blood in stool were done to all samples. K-ras protooncogene mutation at codon 12 (exon 1) and cyclooxygenase 2 (COX-2) -765G>C polymorphism were determined by PCR-RFLP. RESULTS: The K-ras mutation was positive in 23 (28.7%) patients. COX-2 polymorphism revealed GG in 62.5%, GC in 26.2 % and CC genotype was found in 11.3 % of cases. The mean red blood cell folic acid level was lower in the K-ras positive group (100.96±51.3 ng/ml) than the negative group (216.6±166.4 ng/ml), (P<0.01). Higher folate levels were found in males than females (median=173 ng/ml and 85 ng/ml; respectively, P=0.002) with adjusted odds ratio (OR) of 0.984. Only, the RBCs folate (P=0.0018) followed by gender (P=0.036) contributed significantly in the discrimination between patients prone to develop K-ras mutation and those who are not. CONCLUSION: RBC folic acid was significantly deficient in CRC (colorectal cancer) patients with K-ras mutations in comparison with CRC patients free of the mutations, suggesting that folic acid may be a risk factor for K-ras mutation development.

Laparoscopic ovarian drilling in polycystic ovary syndrome: efficacy of adjusted thermal dose based on ovarian volume.

OBJECTIVE: To compare reproductive outcome of adjusted thermal dose on the basis of ovarian volume versus fixed-puncture dosage in laparoscopic ovarian drilling. DESIGN: Randomized controlled trial. SETTING: University Women's Health Center. PATIENT(S): One hundred twenty patients with polycystic ovary syndrome and clomiphene citrate resistance. INTERVENTION(S): Patients were assigned randomly to two groups of 60 women each. Group A received an adjusted thermal dose based on ovarian volume with use of a new model for dose calculation (60 J/cm(3) of ovarian tissue), and group B received 600 J per ovary through four ovarian holes regardless of size. One month afterward, the hormonal profile was reevaluated, and second-look laparoscopy was performed in patients who had not conceived by 6 months to evaluate adnexal adhesions. MAIN OUTCOME MEASURE(S): Ovulation, conception, and early abortion rates, cycle rhythm, and adnexal adhesions. RESULT(S): More patients resumed regular cycles in group A than in group B (87.9% vs. 75.4%). The ovulation and pregnancy rates were significantly higher in group A than in group B (81.8% vs. 62.2% and 51.7% vs. 36.8%, respectively). There was no significant difference between groups in early miscarriage rate or postdrilling adhesions. CONCLUSION(S): Adjusted diathermy dose based on ovarian volume for laparoscopic ovarian drilling of polycystic ovary syndrome has a better reproductive outcome compared with fixed thermal dosage.

Iron chelation therapy in Upper Egyptian transfusion-dependent pediatric homozygous beta-thalassemia major: impact on serum L-carnitine/free fatty acids, osteoprotegerin/the soluble receptor activator of nuclear factor-kappabeta ligand systems, and bone mineral density.

Bone disease in beta-thalassemia major (betaTM) remains poorly understood. Receptor activator of nuclear factor-kappabeta ligand (RANKL) regulates osteoclast formation and function. RANKL activity is balanced by interaction with its receptor (RANK) and binding to osteoprotegerin (OPG). L-Carnitine (LC) enhances osteoblastic activity by furnishing fuel. This study hypothesized that abnormal bone metabolism in betaTM involves imbalanced RANKL/OPG and LC/free fatty acids (FFAs) metabolism. Sixty-nine transfusion-dependent betaTM patients and 15 healthy controls were enrolled. One group of patients (n=34) received desferrioxamine (DFO) and the other (n=35) did not. Serum OPG, soluble RANKL (sRANKL), FFAs, LC [total LC (TC), free LC (FC), and esterified LC (EC)], calcium, and inorganic phosphate were measured by specific immuno and colorimetric assays; bone mineral density was examined by dual x-ray absorptiometry. Patients showed lower levels of OPG, TC, FC, EC and higher levels of sRANKL, sRANKL/OPG ratio, and FFAs than controls. Patients on DFO showed lower levels of OPG, TC, FC and higher levels of sRANKL, sRANKL/OPG ratio, and FFAs than those without chelation. In patients, sRANKL correlated negatively with TC and OPG and FC correlated positively with OPG and negatively with sRANKL, sRANKL/OPG ratio, and FFAs. In conclusion, altered bone metabolism owing to imbalanced osteoclastic bone resorption versus constructive osteoblastic activities in betaTM pediatric patients could be due to abnormal sRANKL-OPG and LC-FFAs systems that were worsened by DFO.