Laila Rashed

BACKGROUND: Alzheimer's dementia (AD) and Parkinsonism are common in geriatric patients. Skeletal muscles are important for proper function of aging animals and humans. This study focuses on the influence of memantine (used in moderate to severe AD) and levodopa/carbidopa (LD/CD) (a cornerstone of Parkinson's disease treatment) on responses of isolated phrenic nerve-diaphragms (IPNDs) of aged male rats.

MATERIAL/METHODS: Of 100 aged male albino rats, 20 were untreated to study the in vitro effects of memantine and LD/CD on IPNDs and 80 were divided into groups 1 (control), 2 (oral memantine, 1.5 mg/kg/d), 3 (twice daily intraperitoneal LD/CD, 25/2.5 mg/kg), and 4 (both drugs). After three weeks-treatment the animals were sacrificed. Ten rats from each group were used to harvest IPNDs to study the effect of gallamine and 10 rats to measure nAchR (nicotinic acetylcholine receptor) alpha subunit mRNA by PCR.

RESULTS: The heights of indirectly elicited contractions were 63.1+/-4.6, 41.5+/-4.5, 70.6+/-4.7, and 53.9+/-3.3 mm for groups 1-4, respectively, and all differences were statistically significant (p<0.05). Memantine caused a leftward shift of the gallamine concentration-response curve and LD/CD a rightward shift. Reversal of neuromuscular block required larger neostigmine concentrations in the memantine group and smaller concentrations in the LD/CD group. In vitro, memantine inhibited diaphragmatic responses to indirect stimulation. Values of nAchR alpha subunit mRNA (microg/dl) were 0.7+/-0.16 (control), 0.13+/-0.11 (memantine), 2.3+/-0.94 (LD/CD), and 1.18+/-0.71 (both drugs) (p<0.05).

CONCLUSIONS: Memantine inhibits neuromuscular transmission in vitro and with in vivo treatment. LD/CD treatment enhances neuromuscular transmission. Clinical implications need further investigation.

UNLABELLED: We investigated the possible role of reactive oxygen species (ROS) on renal function in experimental diabetes.

MATERIALS AND METHODS: Seven groups of male rats were studied. Group I consisted of control animals. Diabetes was induced (by streptozotocin) in the animals in the other groups and they received either insulin or vitamin E (300 or 600 mg/kg), both insulin and vitamin E, or no treatment for 4 weeks. At the end of the study, blood pressure was measured and parameters of kidney function and oxidative stress were evaluated in serum and kidney tissue samples.

RESULTS: Diabetic animals had higher blood pressures; increased serum glucose, urea, creatinine, cyclic guanosine monophosphate (cGMP); increased kidney tissue levels of malondialdehyde and inducible nitric oxide synthetase (iNOS); and reduced serum glutathione peroxidase when compared with control animals. Blood glucose levels in diabetic animals were controlled by insulin and not by any dose of vitamin E alone. However, all other measured parameters improved towards control levels with either insulin or vitamin E in either dose. An additive beneficial effect was observed on the levels of iNOS and cGMP when both forms of treatment were used in diabetic animals.

CONCLUSIONS: We conclude that ROS may play an important role in diabetes-induced nephropathy in this rat model. Vitamin E supplementation in addition to insulin can have additive protective effects against deterioration of renal function in this model.

INTRODUCTION: Erectile response depends on nitric oxide (NO) generated by NO synthase (NOS) enzyme of the nerves and vascular endothelium in the cavernous tissue. NO activates soluble guanylate cyclase (sGC), leading to the production of cyclic guanosine monophosphate (cGMP). cGMP activates cGMP-dependent protein kinase that activates Ca(2+)/ATPase pump that activates Ca(2+)/K efflux pump extruding Ca(2+) across the plasma membrane with consequent smooth muscle cell relaxation. A role similar to that of NOS/NO signaling has been postulated for carbon monoxide (CO) produced in mammals from heme catabolism by heme oxygenase (HO) enzyme.

AIM: To assess CO signaling pathway for erectile function by reviewing published studies.

METHODS: A systematic review of published studies on this affair based on Pubmed and Medical Subject Heading databases, with search for all concerned articles.

MAIN OUTCOME MEASURES: Documentation of positive as well as negative criteria of CO/HO signaling focused on penile tissue.

RESULTS: The concept that HO-derived CO could play a role in mediating erectile function acting in synergism with, or as a potentiator for, NOS/NO signaling pathway is gaining momentum. CO/HO signaling pathway has been shown to partially mediate the actions of oral phosphodiesterase type 5 inhibitors. In addition, it was shown that the use of CO releasing molecules potentiated cavernous cGMP levels. However, increased CO production or release was reported to be associated, in some studies, with vasoconstriction.

CONCLUSION: This review sheds a light on the significance of cavernous tissue CO signaling pathway that may pave the way for creation of therapeutic modalities based on this pathway.

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.

Two major genotypic assemblages of Giardia intestinalis infect humans; the nested real-time polymerase chain reaction (PCR) was used for targeting the triose phosphate isomerase (tpi) gene to detect and genotype G. intestinalis in human feces in Egypt. Among 97 fecal samples, 30 (31%) were diagnosed as giardiasis by saline wet mount microscopy after staining with Lugol's iodine. The tpi gene was amplified from 41 (42.3%) fecal samples, of which 11 were microscopy-negative specimens. Of the total samples, 24 (58.5%) contained assemblage A group I, and 7 (17.1%) were assemblage A group II from the group of patients complaining of intermittent diarrhea. Eight (19.5%) samples contained assemblage B from patients with persistent diarrhea. Two (5%) samples had a mixture of assemblage A group II and assemblage B. The technique was able to detect as few as 20 trophozoites per PCR on fecal DNA-isolated, microscopy-negative, and quantitative (q)PCR-positive specimens; there was a higher average cycle threshold value than microscopy-positive and qPCR-positive specimens, suggesting that they represented true, low-burden infections. In conclusion, we could genotype G. intestinalis from fresh stool samples in Egypt; in infections commonly presented with intermittent diarrhea, the most prevalent genotype was assemblage A group I. The most vulnerable age group included 10- to 20-yr-old individuals.

BACKGROUND: Fibroblast growth factor-23 (FGF23) is a circulating factor that regulates renal reabsorption of inorganic phosphate. Serum FGF23 level is increased in chronic kidney disease (CKD) patients as a compensatory mechanism to hyperphosphataemia. FGF23 directly signals in the parathyroid glands and can be used to predict future secondary hyperparathyroidism in dialysis patients. We examined the relationship between FGF23 and serum calcium, phosphate, 1,25(OH)(2)D(3), and PTH levels in haemodialysis patients.

METHODS: FGF23 and the above mentioned characteristics were measured in 50 chronic haemodialysis patients. We analysed the correlation between FGF23 and the other characteristics by using the Pearson correlation coefficient and multiple regression analysis.

RESULTS: FGF23 was significantly increased in haemodialysis patients compared with healthy controls (1525 +/- 373 vs. 37 +/- 9 pg/ml, P < 0.0001). There was a significant negative correlation between log FGF23 and 1,25(OH)(2)D(3) (R = -0.375, P = 0.009) and a significant positive correlation between log FGF23 and log PTH values (R = 0.287, P = 0.041). In multiple regression analysis log PTH and 1,25(OH)(2)D(3) values were independent predictors of log FGF23 (P = 0.037 and 0.009, respectively).

CONCLUSIONS: Our results revealed a marked increase in FGF23 levels in haemodialysis patients. PTH and vitamin D3 were independent predictors of FGF23 in the study group. Serum phosphate did not correlate with or predict FGF23 level despite the high prevalence of hyperphosphataemia in the study group.

Diabetic nephropathy (DN) is the leading cause of end-stage renal disease in the western world. Increased number of interstitial macrophages has been observed in biopsies from patients with DN. Monocyte chemo-attractant protein-1 (MCP-1) is the strongest known chemo-tactic factor for monocytes and is upregulated in DN. We examined urinary levels of MCP-1 in patients with type-2 diabetes mellitus (DM) to assess its possible correlation with other para-meters of renal injury. The urinary MCP-1 level was assessed in 75 patients with type-2 DM (25 patients each with no microalbuminuria, with macroalbuminuria and, with renal impairment) and compared them with matched healthy control subjects. The HbA1c and estimated glomerular filtration rate (eGFR) derived from the abbreviated Modification of Diet in Renal Disease (MDRD) equation were examined in the study groups in relation to the urinary MCP-1. The urinary MCP-1 level was significantly higher in patients with micro and macroalbuminuria (167.41 +/- 50.23 and 630.87 +/- 318.10 ng/gm creatinine respectively) as compared with normoalbuminuric patients and healthy controls (63.85 +/- 21.15 and 61.50 +/- 24.81 ng/gm creatinine, p p 0.001), HbA1c (r= 0.55, p 0.001) and inversely with eGFR (r=-0.60, p< 0.001). Our findings suggest that hyperglycemia is associated with increased urinary levels of MCP-1 that is closely linked to renal damage as reflected by proteinuria and eGFR levels. Collectively, these findings suggest that MCP-1 is involved in the pathogenesis of diabetic nephropathy through its various stages.

BACKGROUND: The purpose of this study was to investigate the effect of mesenchymal stem cells (MSCs) on cardiovascular complications of type 1 diabetes mellitus (DM) in rats.

MATERIAL/METHODS: MSCs were derived from the bone marrow of male albino rats. The MSCs were characterized morphologically and by RT-PCR for CD29 expression. They were then infused into female rats which were made diabetic by IP injection of streptozotocin (STZ). The rats were divided into control, STZ, and STZ plus MSC groups. Serum insulin, glucose, and fibrinogen were estimated in all groups and the Y-chromosome gene sry was detected by PCR in pancreatic and cardiac tissues. Physiological cardiovascular functions (heart rate, systolic blood pressure) were assessed by a Langendorff apparatus.

RESULTS: Diabetic rats which received MSCs showed significantly lower serum glucose and increased serum insulin levels compared with the STZ group. Improvement of cardiovascular performance was also observed in the STZ/MSC group compared with the STZ group. The sry gene was detected by PCR in the pancreatic and cardiac tissues of the STZ/MSC group.

CONCLUSIONS: Rat bone marrow harbors cells that have the capacity to differentiate into functional insulin-producing cells capable of controlling blood glucose level in diabetic rats. This may provide a source of cell-based therapy for DM. Furthermore, MSC transplantation can improve cardiac function in DM.<

INTRODUCTION: Heme oxygenase (HO) enzyme catalyzes oxidative degradation of heme to biliverdin and carbon monoxide (CO). CO shares many properties with nitric oxide (NO) including the activation of soluble guanyl cyclase.

AIM: To assess cavernous tissue HO activity and cyclic guanosine monophosphate (cGMP) levels in response to oral phosphodiesterase type 5 (PDE5) inhibitors.

METHODS: Seven hundred twenty male Sprague-Dawley rats, divided into six groups, were investigated. Group 1, controls; group 2 received sildenafil citrate orally; group 3 received vardenafil hydrochloride; and group 4 received tadalafil. Group 5 was subdivided into three equal subgroups, received the same dose of each drug added to the HO inhibitor, Zn protoporphyrin. Group 6 was subdivided into three equal subgroups, received the same dose of each drug added to the NO inhibitor, L-nitroarginine methylester. Eight rats from each group/subgroup were sacrificed at 0.5, 1, 2, 3, 4, 6, 18, 24, and 36 hours, respectively.

MAIN OUTCOME MEASURES: HO enzyme activity assay and cGMP tissue levels in dissected rat cavernous tissues.

RESULTS: Both cavernous tissue HO enzyme activity and cGMP levels were increased significantly in sildenafil-, vardenafil-, and tadalafil-treated rats compared with the controls, with significant decreases after either HO or NO inhibition. Cavernous tissue HO enzyme activity and cGMP showed a positive significant correlation (r = 0.854, P < 0.001).

CONCLUSION: The effects of PDE5 inhibitors in cavernous tissue are partly mediated through HO enzyme activity.

INTRODUCTION: Cyclic guanosine monophosphate (cGMP) levels can be regulated by heme oxygenase-1 and 2 (HO-1 and HO-2)-derived carbon monoxide (CO).

AIMS: Assessment of the effect of upregulating CO in rat corpora cavernosa (CC) on cavernous cGMP.

METHODS: Three experimental groups were studied: first group (N = 40), short-term HO induction over 2 weeks by injection of intraperitoneal increasing doses of hemin; the second group (N = 40) was subjected to intracavernosal injection of CO donor, CORM-3, or its inactive form (iCORM-3) over 2 weeks; the third group (N = 60) was subdivided into three subgroups: the first one received a combined hemin and CORM-3, the second one received hemin and its inhibitor stannus mesoporphyrin (SnMP), and third one received a combined hemin, CORM-3, and SnMP.

MAIN OUTCOME MEASURES: In CC, HO-1 and HO-2 gene expression, Northern blot and Western blot, cGMP levels, and HO enzyme activity.

RESULTS: In the first group, maximum induction of HO-1 gene expression, HO enzyme activity, and cGMP occurred with 4-mg hemin dose with a successive increase over 2 weeks. In the second group, CORM-3 increased cGMP by twofold compared with iCORM-3, and also increased HO-1 protein. In the third group, SnMP inhibited the enhancing effect of CORM-3 and HO on erectile signaling molecules; i.e., HO-1 gene, enzyme activity, and cGMP.

CONCLUSIONS: CORM-3- or hemin-mediated CO release could increase cavernous tissue cGMP.

The protein kinase C (PKC) family consists of 13 members categorized as conventional or novel depending on whether diacylglycerol, calcium, or phosphatidylserine is required for activation. High glucose leads to activation of different forms of PKC across tissue types, thus determining the kind of diabetes-induced organ damage. PKC beta was reported to have a positive role in B-lymphocyte activity through activation of NF-kB, leading to various immune disorders. We examined renal expression of two PKC isoforms alpha and beta in renal biopsies of patients with diabetic nephropathy, lupus nephritis (LN) (Class 3-4), and mesangioproliferative glomerulonephritis (MPGN) to explore the role of each isoform in different glomerular diseases. PKC alpha and beta gene expression was studied by quantitative real-time reverse transcription-PCR in 20 patients with type 2 diabetes and proteinuria (serum creatinine 2.04 +/- 0.85 mg/dl, 24-h urinary protein 3.61 +/- 1.75 g, eGFR 37.85 +/- 17.89 ml/min/1.73 m2), 20 patients with proliferative LN (serum creatinine 1.67 +/- 1.50 mg/dl, 24-h urinary protein 4.46 +/- 5.01 g, eGFR 69.62 +/- 40.93 ml/min/1.73 m2), and 20 patients with MPGN (serum creatinine 3.32 +/- 2.79 mg/dl, 24-h urinary protein 4.65 +/- 4.11 g, eGFR 32.62 +/- 29.56 ml/min/1.73 m2). Normal tissues from the normal pole of four kidneys removed because of renal tumor served as controls. PKC á gene expression was significantly increased in diabetic kidneys compared to LN and MPGN (316.95 +/- 152.94 microg/ml vs. 185.97 +/- 32.13 and 195.46 +/- 46.45 microg/ml, p < 0.05). PKC â gene expression was significantly increased in the LN and MPGN groups compared to the diabetic nephropathy group (41.01 +/- 14.03 and 39.93 +/- 16.41 microg/ml, respectively, vs. 18.20 +/- 4.91 microg/ml, p < 0.05). Significant correlation was noted between the PKC alpha gene concentrations and proteinuria in diabetic patients. Renal expression of PKC alpha and beta genes in control tissues were significantly lower compared to diabetic kidneys, LN, and MPGN groups (32.31 +/- 0.36 and 4.67 +/- 2.41 microg/ml, respectively, p < 0.001). The study revealed enhanced renal gene expression of both PKC isoforms alpha and beta in diabetic kidney tissues, LN, and MPGN, but in different patterns. PKC alpha gene expression was significantly increased in diabetic patients with chronic kidney disease. The increased expression of the PKC beta gene in LN and MPGN highlights its role in regulation of the immune system. This may represent potential therapeutic targets for prevention of progressive kidney injury in diabetic and proliferative glomerular diseases.

AIM: To investigate the possible role of oxidative stress as a common mediator of apoptosis and cardiac damage in diabetes.

MATERIALS AND METHODS: This experimental work was conducted on 5 groups of Wistar rats. Group I was the control group. Diabetes type 1 was induced in other groups (by streptozotocin) and animals received insulin or vitamin E (300 mg /kg body weight), both insulin and vitamin E, or no treatment for 4 weeks according to their group. At the end of the study, serum and cardiac tissues were examined for biochemical parameters of cardiac function, oxidative stress and apoptosis. Electron microscopy pictures of cardiac tissue were also evaluated for signs of cardiac damage

RESULTS: Markers of oxidative stress, apoptosis, inflammation as well as manifestations of cardiac damage as assessed by electron microscopy were significantly decreased in rats treated with both insulin and vitamin E when compared with untreated diabetic rats or rats treated with either insulin or vitamin E alone

CONCLUSION: Administration of both vitamin E and insulin was effective in reducing markers of oxidative stress and apoptosis and improving parameters of cardiac function in experiments animals. Antioxidants might prove beneficial as an adjuvant treatment in addition to insulin in type 1 diabetes associated with manifestations of cardiac complications.

AIM: To assess heme oxygenase-1 (HO-1) activity in the cavernous tissue of sildenafil citrate-treated rats.

METHODS: One hundred and ninety-two Sprague-Dawley male rats, divided into four equal groups, were investigated. Group 1, the control group, received regular animal chow; group 2 received sildenafil citrate by intragastric tube; group 3 received sildenafil and HO inhibitor (zinc protoporphyrin, ZnPP); and group 4 received sildenafil and nitric oxide synthase (NOS) inhibitor L-nitroarginine methyl ester (L-NAME). Twelve rats from each group were killed after 0.5 h, 1 h, 2 h and 3 h of drug administration. Then HO-1 activity, cGMP levels and NOS enzymatic activity in the cavernous tissues were estimated.

RESULTS: In cavernous tissue, HO-1 activity, NOS enzymatic activity and cGMP concentration increased significantly in sildenafil-treated rats compared to other groups throughout the experiment. Rats receiving either HO or NOS inhibitors showed a significant decrease in these parameters. HO-1 cavernous tissue activity and NOS enzymatic activity demonstrated a positive significant correlation with cGMP levels (r = 0.646, r = 0.612 respectively; P < 0.001).

CONCLUSION: The actions of PDE5 inhibitor sildenafil citrate in the cavernous tissue are partly mediated through the interdependent relationship between both HO-1 and NOS activities.

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.

A total of 140 out of 180 outpatients attended MISR University for Science and Technology Hospital complained of abdominal pain, diarrhoea and/or dysentery. Stool examination showed 47 (33.6%) had Entamoeba sp., 36 (25.7%) had cysts and 11 (7.9%) had trophozoites. Of 40 asymptomatic ones, 4 (10%) had cysts. A total of 51 positive stool samples for Entamoeba sp. (40 cysts & 11 trophozoites) were tested by Ne-sted Polymerase Chain Reaction (N-PCR) and Restriction Enzyme Digestion (RED) to clarify true E. histolytica from E. dispar. The results showed that 9/51 (17.6%) had E. dispar, while 31 (60.8%) had E. histolytica and 11 (21.6%) had dual infection with both E. histolytica and E. dispar. All E. histolytica PCR proved cases were from the symptomatic group, 11 had trophozoites and 34 had cysts. Thus, the result showed the potential use of molecular tools in detection of E. histolytica and E. dispar, and is a promising tool for epidemiology, particularly to differentiate pathogenic and non pathogenic Entamoeba sp.

OBJECTIVE: To evaluate the serum transforming growth factor-beta 1 level in type II diabetic patients with diabetic nephropathy (DN) and to assess its use as a marker of renal injury in type II diabetes.

METHODS: Sixty patients with type II diabetes mellitus (DM), who attended the outpatient internal medicine clinic in Cairo University Hospital, Egypt from January 2003 to March 2003, were the subjects of the present study and compared to 10 healthy age- and gender- matched control subjects They were divided into 6 major groups according to degree of metabolic control, as determined by glycosylated hemoglobin (HbA1c), the rate of urinary albumin excretion (UAE) and serum creatinine level. Serum transforming growth factor-beta 1 level was assessed by enzyme linked immunosorbent assay (ELISA).

RESULTS: Serum transforming growth factor-beta 1 level was significantly increased in micro albuminuric (UAE 20-200 ug/minute), macro albuminuric (UAE >200 ug/minute) and overtly nephropathic diabetic patients with renal impairment compared to healthy controls (p<0.05). In addition, serum transforming growth factor-beta 1 level was significantly increased in type II diabetic patients with poor glycemic control (HbA1c >7.6%) compared to patients with good glycemic control (HbA1c 5.5-7.6%). Serum transforming growth factor-beta 1 level was significantly increased in hypertensive DM patients compared to normotensive DM patients (p<0.05). There was a strong correlation between serum transforming growth factor-beta 1 level and HbA1c, blood urea, serum creatinine and 24-hour urinary protein excretion (p<0.01).

CONCLUSION: Our data strongly support the hypothesis that hyperglycemia may trigger the activation of transforming growth factor-beta 1 which in turn mediates progressive renal damage in type II DM. Increased serum transforming growth factor-beta 1 may be useful as a marker of diabetic renal disease as it shows a close association with the parameters of renal injury in type II diabetes.

Forty nine stool specimens collected from severe diarrheic patients. Eight were suffering from Hodgkin's lymphoma, and the rest were suffering from acute lymph plastic leukaemia. All were examined microscopically for protozoan parasites mainly, Cryptosporidium parvum and Cyclospora cayetanensis. Of the patients, 34 (69.4%) were positive and 15 (30.6%) were negative by both microscopy and nested PCR. An additional 12 (24.5%) who were negative by microscopy were positive by nested PCR. Stool examination revealed 16 cases with C. parvum, and 6 with C. cayetanensis, and 3 cases showed mixed infection. The results were compared with the established nested PCR assay to detect DNA directly from stool specimens. The patients <3 years old more affected by Cryptosporidium infection, unlike Cyclospora sp. Infection was in older age groups, which reflected the modes of parasite' transmission.. Diarrheal illness was stronger for Cyclospora than for Cryptosporidium. After the extraction of DNA from stool, a 402-bp fragment of C. parvum, and 602 bp fragment of C. cayetanensis was amplified. The amplified products, 194-bp DNA fragment for C. parvum, and 306 bp DNA fragment of C. cayetanensis were used for the second run. This study indicated that primers were specific for DNA of C. parvum and C. cayetanensis. PCR detected a total of 22 (44.9%) positives for C. parvum infection (6 negative by AF stool examination), and 12 (24.5%) positives for C. cavetanensis. Infection (6 negative by AF stool examination), 7 (14.3%) showed mixed infection (4 negative by AF stool examination), all microscopic negative specimens were positive by successive stool examination. Microscopy exhibited sensitivity of 72.7% for C. parvum, 50% for C. cayetanensis and 100% specificity for both parasites compared to 100% sensitivity and specificity with PCR. So, PCR is more sensitive and easier to interpret but required more hands-on time to perform and is more expensive. However, PCR batch analysis reduces the cost considerably.

Retrovirus-mediated interferon alpha (IFN-alpha) gene transfer was evaluated with regard to its possible protective effects against aflatoxin B1 (AFB1)-initiated and carbon tetrachloride (CCl4)-promoted hepatic carcinogenesis in rats. To our knowledge, this is the first time an experimental in vivo gene therapy trial was conducted in Egypt. Two genes were examined in liver tissue by RT-PCR: the first was glutathione-S-transferase placental (GST-P) isoenzyme, as an early marker to detect hepatic malignancy; the second was IFN-alpha gene expression to detect the efficiency of gene uptake and its persistence after transduction. Forty male rats, divided equally into 4 groups, were included in the study: the first group was the control; the second group received CCl4 0.2 ml subcutaneously twice weekly for 12 weeks and AFB1 0.25 mg/kg body wt intraperitoneally twice weekly for 6 weeks; the third group received IFN-alpha (10(8) pfu) intravenously in the tail vein prior to the start of CCl4 and AFB1 injections; and the fourth group received IFN-alpha (10(8) pfu) by intrahepatic injection under ultrasonography guide after termination of the CCl4 and AFB1 injection schedule. The results showed that IFN-alpha has a marked and significant protective effect against hepatic fibrogenesis as well as hepatic carcinogenesis. Pathological examination of liver tissue proved that IFN-alpha minimized both fibrotic and cirrhotic processes. The amount of fibrosis was less in both groups receiving IFN-alpha, with more protection in the group that received IFN-alpha intravenously prior to CCl4 and AFB1. The results of RT-PCR showed that the IFN-alpha gene was significantly expressed in both groups receiving IFN-alpha, with a more intense expression in the group that received IFN-alpha by intrahepatic injection after termination of CCl4 and AFB1 injections. The IFN-alpha gene was detected after three months of gene transduction in rats receiving IFN-alpha intravenously prior to CCl4 and AFB1 and after one month of gene transduction in the post CCl4 and AFB1 rats. IFN-alpha gene was not expressed in the two groups that did not undergo gene transfer. Histopathological signs of premalignant macronodules were evident in the group receiving CCl4 and AFB1, but not IFN-alpha as well as in the group that received IFN-alpha at the end of the experiment. GST-P gene expression was also detected in these two groups, confirming early malignant transformation. In conclusion, IFN-alpha exerts significant protective effects, but more so when the gene is administered before fibrogenic and carcinogenic induction in hepatic tissues. IFN-alpha gene therapy may be justified in clinical trials for high-risk candidates with hepatic carcinogenesis.

To investigate the immunomodulatory effect of the Th1 mediated cytokine IFN-alpha on schistosomiasis, this cytokine was weekly injected into mice experimentally infected with S. mansoni, beginning from day 0 (group II), week 3 (group III), week 6 (group IV) and week 10 (group V) post-infection. TGF-beta1 serum levels were estimated on a weekly basis and beginning one week after initiation of IFN-alpha therapy, while all animals were sacrified on week 14 to be used for egg counts in liver and small intestine, oogram study for determination of the maturity of deposited eggs, and histopathological examination of stained liver sections. IFN-alpha treated groups were characterized by a more intense oviposition in the intestine (liver/intestine ratio less than 1), with higher egg numbers the earlier IFN-alpha was administered. Oograms of the intestine indicated the level of immature eggs to be statistically significantly higher in group II, III and IV than in the control group I (p < 0.05). In IFN-alpha medicated mice, the mean numbers and diameters of hepatic granulomas were less than in GI, in addition to a lower representation of fibrocellular and fibrous granulomas among them (all parameters p < 0.05), especially in Gs IV & V. The inflammatory cell population in the form of eosinophils, histiocytes and giant cells was more pronounced in Gs III, IV & V. TGF-beta1 serum levels showed a progressive rise, however more pronounced in the untreated control. A statistically positive significant was established between TGF-beta1 levels and number, size and percentage of fibrotic hepatic granulomas in all groups.

Fifty stool specimens collected from severe diarrheic patients attending Misr University Hospital, were examined microscopically for protozoan parasites mainly, Cryptosporidium parvum. Stool examination revealed 22 cases with C. parvum, 8 with E. histolytica, 14 with G. intestinalis and six were parasite-free. The results were compared with the established nested PCR assay to detect DNA directly from stool specimens. After the extraction of DNA from stool, a 402-bp fragment of C. parvum DNA was amplified with two 26-mer outer primers. The amplified products, 194-bp DNA fragment, were used for a second run. This study indicated that the used primers are specific for DNA of C. parvum. The PCR detected a total of 28 positives; six of these cases were negative by AF stool examination, which eventually confirmed to be positive by several successive examinations of the stool and/or duodenal aspiration. Microscopy exhibited 78.5% sensitivity and 100% specificity compared to 100% specificity and sensitivity with PCR. Consequently, PCR is more sensitive and easier to interpret but required more hands-on time to perform and is more expensive than microscopy. However, PCR batch analysis reduces the cost considerably.