Mahmoud Abd El Hafez M. Adam

AIMS AND BACKGROUND: This study aims to detect the expression of E-selectin in tissue and the serum level of its soluble form in patients with primary breast cancer and benign breast tumors and to correlate the results with the clinicopathological data of the subjects.

METHODS: Fifty participants were included in the study and stratified into 3 subgroups. Group A comprised 30 patients with primary breast cancer, group B 9 patients with benign breast tumors, and group C 11 healthy control women undergoing reduction mammoplasty. E-selectin gene expression was investigated in breast tissues by PCR techniques and soluble E-selectin was measured in sera by ELISA.

RESULTS: The E-selectin gene was expressed in 73.3% of group A, 44.4% of group B and 9.1% of group C. It was expressed in 61.5% of patients with grade 2 breast cancer and in 82.4% of patients with grade 3 breast cancer. E-selectin gene expression was detected in 60%, 73.3% and 100% of patients with stage II, III and IV tumors, respectively. It was detected in 81.8% of patients with node-positive primary breast cancer and in 50% of patients with node-negative cancer. PCR in situ hybridization was done to locate the site of E-selectin expression. E-selectin was found on the membranes of peritumoral endothelial cells while it was not found on breast epithelial cells. Serum levels of soluble E-selectin were significantly elevated in group A compared to groups B and C (P < 0.001). They increased significantly with increasing breast cancer stage (P < 0.001) and were significantly higher in patients with lymph node involvement than in patients without node involvement (P < 0.001).

CONCLUSIONS: The studied marker showed associations with established prognostic parameters such as lymph node involvement and histological tumor grade. Further studies are needed to evaluate E-selectin as a possible target for antimetastatic therapy through modulation of the expression of the cell adhesion molecule. E-selectin can be regarded as a promising strategy in improving tumor therapy.

Mycobacterium ulcerans and Mycobacterium marinum are closely related pathogens which share an aquatic environment. The pathogenesis of these organisms in humans is limited by their inability to grow above 35 degrees C. M. marinum causes systemic disease in fish but produces localized skin infections in humans. M. ulcerans causes Buruli ulcer, a severe human skin lesion. At the molecular level, M. ulcerans is distinguished from M. marinum by the presence of a virulence plasmid which encodes a macrolide toxin, mycolactone, as well as by hundreds of insertion sequences, particularly IS2404. There has been a global increase in reports of fish mycobacteriosis. An unusual clade of M. marinum has been reported from fish in the Red and Mediterranean Seas and a new mycobacterial species, Mycobacterium pseudoshottsii, has been cultured from fish in the Chesapeake Bay, United States. We have discovered that both groups of fish pathogens produce a unique mycolactone toxin, mycolactone F. Mycolactone F is the smallest mycolactone (molecular weight, 700) yet identified. The core lactone structure of mycolactone F is identical to that of M. ulcerans mycolactones, but a unique side chain structure is present. Mycolactone F produces apoptosis and necrosis on cultured cells but is less potent than M. ulcerans mycolactones. Both groups of fish pathogens contain IS2404. In contrast to M. ulcerans and conventional M. marinum, mycolactone F-producing mycobacteria are incapable of growth at above 30 degrees C. This fact is likely to limit their virulence for humans. However, such isolates may provide a reservoir for horizontal transfer of the mycolactone plasmid in aquatic environments.

Heparan sulfate (HS), is a proteoglycan (PG) found both in the extracellular matrix and on cell surface. It may represent one of the most biologically important glycoconjugates, playing an essential role in a variety of different events at molecular level. The publication of the mouse genome, and the intensive investigations aimed at understanding the proteome it encodes, has motivated us to initiate studies in mouse glycomics focused on HS. The current study is aimed at determining the quantitative and qualitative organ distribution of HS in mice. HS from brain, eyes, heart, lung, liver, kidney, spleen, intestine and skin was purified from 6-8 week old male and female mice. The recovered yield of HS from these organs is compared with the recovered whole body yield of HS. Structural characterization of the resulting HS relied on disaccharide analysis and (1)H-NMR spectroscopy. Different organs revealed a characteristic HS structure. These data begin to provide a structural understanding of the role of HS in cell-cell interactions, cell signaling and sub-cellular protein trafficking as well as a fundamental understanding of certain aspects of protein-carbohydrate interactions.

The homogeneity and stability of the static magnetic field are of paramount importance to the accuracy of MR procedures that are sensitive to phase errors and magnetic field inhomogeneity. It is shown that intense gradient utilization in clinical horizontal-bore superconducting MR scanners of three different vendors results in main magnetic fields that vary on a long time scale both spatially and temporally by amounts of order 0.8-2.5 ppm. The observed spatial changes have linear and quadratic variations that are strongest along the z direction. It is shown that the effect of such variations is of sufficient magnitude to completely obfuscate thermal phase shifts measured by proton-resonance frequency-shift MR thermometry and certainly affect accuracy. In addition, field variations cause signal loss and line-broadening in MR spectroscopy, as exemplified by a fourfold line-broadening of metabolites over the course of a 45 min human brain study. The field variations are consistent with resistive heating of the magnet structures. It is concluded that correction strategies are required to compensate for these spatial and temporal field drifts for phase-sensitive MR protocols. It is demonstrated that serial field mapping and phased difference imaging correction protocols can substantially compensate for the drift effects observed in the MR thermometry and spectroscopy experiments.

The TALLYHO/JngJ (TH) strain is a newly established, polygenic mouse model for type 2 diabetes (T2D) and obesity, and we have previously reported some key physiological features of this model after the overt onset of diabetes. In the present work, we conducted a comprehensive phenotypic characterization of TH in order to completely characterize this new and relevant model for human T2D and obesity. We monitored the development of obesity and diabetes starting at 4 weeks of age by measuring body weight, glucose tolerance, and plasma levels of insulin, glucose, and triglyceride. Additionally, histological alterations in the pancreas and glucose uptake and glucose transporter 4 (GLUT4) content in soleus muscle were also examined. Compared with age- and sex-matched C57BL/6J (B6) mice, both male and female TH mice were significantly heavier, hyperleptinemic, and hyperinsulinemic at 4 weeks of age, without glucose intolerance or hyperglycemia. TH mice maintained higher body weights throughout the study period of 16 weeks. The hyperinsulinemia in TH mice worsened with age, but to a lesser degree in females than in males. Both the male and the female TH mice had enlarged pancreatic islets. Male TH mice showed impaired glucose tolerance at 8 weeks that became more prominent at 16 weeks. Plasma glucose levels continuously increased with age in male TH mice resulting in frank diabetes, while female TH mice remained normoglycemic throughout the study. Impaired glucose tolerance and hyperglycemia in male TH mice were accompanied by impaired 2-deoxyglucose uptake in the soleus muscle at basal and insulin-stimulated states, but without any reduction in GLUT4 content. Interestingly, male TH mice exhibited a drastic elevation in plasma triglyceride levels in the pre-diabetic stage that was maintained throughout the study. These findings suggest that obesity and insulin resistance are an inherent part of the TH phenotype and glucose intolerance is evident preceding progression to overt diabetes in male TH mice.

Lipoteichoic acids (LTAs) are Gram-positive bacterial cell wall components that elicit mononuclear cell cytokine secretion. Cytokine-stimulating activity is thought to be dependent on retaining a high level of ester-linked D-alanine residues along the polyglycerol phosphate backbone. However, Streptococcus pyogenes LTA essentially devoid of D-alanine caused human and mouse cells to secrete as much IL-6 as LTA with a much higher D-alanine content. Furthermore, hemoglobin (Hb) markedly potentiates the stimulatory effect of various LTAs on mouse macrophages or human blood cells, regardless of their d-alanine content. LTA and Hb appear to form a molecular complex, based on the ability of each to affect the other's migration on native acrylamide gels, their comigration on these gels, and the ability of LTA to alter the absorption spectra of Hb. Because S. pyogenes is known to release LTA and secrete at least two potent hemolytic toxins, LTA-Hb interactions could occur during streptococcal infections and might result in a profound alteration of the local inflammatory response.

Nitric oxide (NO) plays an important role in anoxic preconditioning to protect the heart against ischemia-reperfusion injuries. The present work was performed to study better the NO-cGMP-protein kinase G (PKG) signaling pathway in the activation of both sarcolemmal and mitochondrial ATP-sensitive K+ (KATP) channels during anoxic preconditioning (APC) and final influence on reducing anoxia-reperfusion (A/R)-induced cardiac damage in rat hearts. The upstream regulating elements controlling NO-cGMP-PKG signal-induced KATP channel opening that leads to cardioprotection were investigated. The involvement of both inducible and endothelial NO synthases (iNOS and eNOS) in the progression of this signaling pathway was followed. Final cellular outcomes of ischemia-induced injury after different preconditioning in the form of lactate dehydrogenase release, DNA strand breaks, and malondialdehyde formation as indexes of cell injury and lipid peroxidation, respectively, were investigated. The lactate dehydrogenase and malondialdehyde values decreased in the groups that underwent preconditioning periods with specific mitochondrial KATP channels opener diazoxide (100 microM), nonspecific mitochondrial KATP channels opener pinacidil (50 microM), S-nitroso-N-acetylpenicillamine (SNAP, 300 microM), or beta-phenyl-1,N2-etheno-8-bromoguanosine-3',5'-cyclicmonophosphorothioate, Sp-isomer (10 microM) before the A/R period. Preconditioning with SNAP significantly reduced the DNA damage. The effect was blocked by glibenclamide (50 microM), 5-hydroxydecanoate (100 microM), NG-nitro-L-arginine methyl ester (200 microM), and beta-phenyl-1,N2-etheno-8-bromoguanosine-3',5'-cyclic monophosphorothioate, Rp-isomer (1 microM). The results suggest iNOS, rather than eNOS, as the major contributing NO synthase during APC treatment. Moreover, the PKG shows priority over NO as the upstream regulator of NO-cGMP-PKG signal-induced KATP channel opening that leads to cardioprotection during APC treatment.

PURPOSE: We present a series of 400 patients with tumor in a solitary kidney who underwent open surgical partial nephrectomy performed by a single surgeon (ACN) with a primary focus on postoperative long-term kidney function.

MATERIALS AND METHODS: A total of 400 patients with sporadic nonfamilial kidney tumors in a solitary kidney underwent open partial nephrectomy between 1980 and 2002. In 323 patients (81%) the contralateral kidney had been surgically removed, while the remaining 77 (19%) had a congenital solitary kidney. Renal insufficiency was present preoperatively in 184 patients (46%). Adverse risk factors for partial nephrectomy were present in a large percent of patients. Intraoperative and postoperative parameters were evaluated at a mean followup of 44 months.

RESULTS: In the overall series 5 and 10-year cancer specific survival was 89% and 82%, respectively. Surgical complications occurred in 52 patients (13%), most commonly urinary leakage. Early postoperative renal function was achieved in 398 patients (99.5%). Only 2 patients required permanent dialysis postoperatively. Satisfactory long-term renal function was achieved in 382 patients (95.5%). A total of 18 patients had progressed to renal failure a mean of 3.6 years after surgery. Patient age, the amount of renal parenchyma resected, a congenitally absent or atrophic contralateral kidney and the time of contralateral nephrectomy were noted to be significantly associated with postoperative renal function.

CONCLUSIONS: Open surgical partial nephrectomy can be safely performed in patients with tumor in a solitary kidney. Long-term cancer-free survival with the preservation of renal function can be reliably expected in most of these cases.

We examined the effects of the protein kinase A (PKA) inhibitor H-89 on voltage-dependent K(+) (K(V)) currents in freshly isolated rabbit coronary arterial smooth muscle cells, using a whole-cell patch clamp technique. H-89 inhibited the K(V) current in a concentration-dependent manner, with a K(d) value of 1.02 microM. However, the PKA inhibitors KT 5720 and Rp-8-CPT-cAMPS did not significantly alter the K(V) current or the inhibitory effects of H-89 on the K(V) current. Moreover, H-85, a structurally similar but inactive analog of H-89, showed similar inhibitory effects on the K(V) channel. H-89 had no effect on the voltage-dependency of activation or inactivation, or on recovery kinetics. These results suggest that in rabbit coronary arterial smooth muscle cells, H-89 inhibits the K(V) current directly by blocking the pore cavity, an effect independent of PKA inhibition.

We investigated the effects of the vasoconstrictor angiotensin (Ang) II on the whole cell inward rectifier K(+) (Kir) current enzymatically isolated from small-diameter (<100 microm) coronary arterial smooth muscle cells (CASMCs). Ang II inhibited the Kir current in a dose-dependent manner (half inhibition value: 154 nM). Pretreatment with phospholipase C inhibitor and protein kinase C (PKC) inhibitors prevented the Ang II-induced inhibition of the Kir current. The PKC activator reduced the Kir currents. The inhibitory effect of Ang II was reduced by intracellular and extracellular Ca(2+) free condition and by Gö6976, which inhibits Ca(2+)-dependent PKC isoforms alpha and beta. However, the inhibitory effect of Ang II was unaffected by a peptide that selectively inhibits the translocation of the epsilon isoform of PKC. Western blot analysis confirmed that PKCalpha, and not PKCbeta, was expressed in small-diameter CASMCs. The Ang II type 1 (AT(1))-receptor antagonist CV-11974 prevented the Ang II-induced inhibition of the Kir current. From these results, we conclude that Ang II inhibits Kir channels through AT(1) receptors by the activation of PKCalpha.

INTRODUCTION: Fracture of the penis is a condition where excessive force applied to the long axis of the penis in the erect state results in rupture of the tunica albuginea of the corpus cavernosum. Surgical management can be confusing and time-consuming due to the concealment of the tear in organized blood and edematous tissue, necessitating extensive dissection in friable traumatized tissues, especially if the tear is a small one, or if there are multiple tears.

AIM: The present work investigates the value of methylene blue in aiding the localization of tunical and urethral tears in such cases.

PATIENTS AND METHODS: Twelve cases with delayed presentation of fracture penis were managed. In six patients, methylene blue was injected into the corpora cavernosa and through the urethral meatus to point out tears. In the other six patients, methylene blue was not used.

MAIN OUTCOME MEASURES: Operative time and complication rate.

RESULTS: Operative time was considerably less in the group that received methylene blue, and the repair was more straightforward. Complications issued only in the group that did not receive methylene blue considering the extensive lengthy dissection.

CONCLUSION: Methylene blue-guided repair for trauma of the penis is an easy, reliable, safe, and fast method for spotting tears in the tunica albuginea of the corpora cavernosa or in the urethra, eliminating the need for unnecessary lengthening.

Four water buffalo calves (Bubalus bubalis) were each inoculated intravenously with 10(6)T. evansi (camel isolate) and the fifth calf kept as non-infected control. The blood and sera of all calves were examined every 4 days during the first month post-inoculation (pi) and then once weekly until the end of the experiment (88 days pi). They were examined for hematological and biochemical changes, liver and kidney function tests. Hemoglobin concentration (Hb%), packed cell volume (PCV) and red blood cell count were significantly decreased. Total leucocytic count, lymphocytes and monocytes showed significant increase. Liver function tests revealed significant elevation in the activity of lactate dehydrogenase enzyme (LDH), globulin, total biliruben and indirect biliruben while alkaline phosphatase enzyme showed significant decrease. Kidney function tests revealed significant decrease of both creatinine and urea.

BACKGROUND: Inadequate cyclosporine (CsA) blood levels are a major risk factor for acute rejection in transplant recipients. The CsA trough level (C0 level) measured just before the next dose is commonly used to adjust the oral dosage. However, the 2-hour post-CsA dose concentration (C2 level) is favored as the best single-point correlate of CsA area-under-the-curve concentration and may better reflect the immunosuppressive effect of CsA. Because an adequate C2 level has not yet been defined, this study was performed to assess the value of C2 monitoring for the prevention of acute rejection and to define target levels in pediatric heart transplant recipients.

METHODS: C2 levels were assessed in 50 pediatric heart transplant patients with oral CsA therapy and compared with trough C0 levels using full blood sampling, mass spectrometry and a blinded analysis. Acute graft rejection was detected using intramyocardial electrocardiogram (IMEG) and serial conventional and tissue Doppler echocardiography (TDE). Rejection was confirmed or excluded by endomyocardial biopsy.

RESULTS: C2 and not C0 levels were significantly reduced in patients with acute graft rejection (ISHLT Grade > or =2). Patients with a C2 level <600 ng/ml had a significantly higher risk of developing acute rejection (100% sensitivity and 82% specificity). Patients with impaired CsA absorption were identified with C2 monitoring and switched to another calcineurin inhibitor.

CONCLUSIONS: Monitoring of the C2 rather than the C0 level better reflects immunosuppressive efficiency and identifies patients at increased risk of acute rejection. A C2 level of >600 ng/ml should be the target to prevent acute rejection.

PURPOSE: The particular goal of this work is to study some cell cycle regulatory proteins and their potential impact on prognosis of breast cancer; p53, cyclin D1 and p27 are potential effectors being the major contributors to the control of the restriction (R) check point of the cell cycle. We also aimed to evaluate different techniques used to detect these cell cycle proteins.

MATERIAL AND METHODS: Forty five breast cancer patients as well as 10 controls with non malignant pathology were assessed for cell cycle regulators each by 2 different techniques; p53 was assessed by enzyme immunoassay (EIA) and immunohistochemistry (IHC), cyclin D1 by Western Blotting (WB) and IHC and p27 by WB and IHC. The cut-off was calculated as the mean of the normal controls +2 SD. Patients were followed up for 4 years and their laboratory data were correlated with different clinical parameters and with other studied regulators.

RESULTS: Eighty seven percent of cases (39/45) were positive for p53 by EIA with a range from 20 to 4300, and a mean of 464 +/- 971 pg/mg protein. By IHC, 80% (24/30) of the cases showed varying degrees of positivity. Using WB, cyclin D1 showed high expression levels above cut off values in 69% of patients (31/45) and in 67% (20/30) by IHC. The corresponding positive figures for p27 were 82% (37/45) and 73% (22/30) using the two techniques, respectively. No significant association was found between p53, cyclin D1 and p27 on one side and different clinical parameters as lymph node status, tumor size or presence of distant metastases on the other side. Survival was poor in patients with high p53 expression. Cyclin D1 positive cases showed comparable survival with negative cases, whereas high p27 levels favored a longer disease free survival.

CONCLUSIONS: Techniques more suitable for assessment of each of these markers in our consideration were EIA for p53, WB for cyclin D1 and IHC for p27. Moreover, this study demonstrated that these markers were relevant to the biological behavior of the tumor cell per se with a possible impact on prognosis and survival, independent of other clinical prognostic factors.

BACKGROUND: Rotavirus is a leading cause of morbidity and mortality in children younger than 5 years, but there is no effective treatment. We assessed the activity of nitazoxanide, a broad-spectrum anti-infective drug, against rotavirus in cell culture and in a clinical trial in paediatric patients hospitalised with severe rotavirus diarrhoea.

METHODS: We did a randomised double-blind placebo-controlled trial in 50 children admitted to the Cairo University Children's Hospital between June 15 and Aug 23, 2005, with severe rotavirus diarrhoea. 38 children aged 5 months to 7 years (median age 11 months) with rotavirus as the sole identified cause of gastroenteritis were enrolled in the clinical study. Patients were randomly assigned either 7.5 mg/kg nitazoxanide as an oral suspension or placebo twice a day for 3 days, and all remained in hospital for 7 days after start of treatment. The primary endpoint was time from first dose to resolution of illness, and analysis was by modified intention-to-treat. This study is registered with ClinicalTrials.gov, number NCT00302640.

FINDINGS: Survival analysis showed that the median time to resolution of illness was 31 h (IQR 22-73) for the nitazoxanide-treated group compared with 75 h (51-124) for the placebo group (p=0.0137). No significant adverse events were reported.

INTERPRETATION: A 3-day course of nitazoxanide significantly reduced the duration of rotavirus disease in hospitalised paediatric patients. These results are encouraging, and might lead us to think about new approaches to managing rotavirus disease in children.

Diabetes represents a serious risk factor for the development of cardiovascular problems such as coronary heart disease, peripheral arterial disease, hypertension, stroke, cardiomyopathy, nephropathy and retinopathy. Identifying the pathogenesis of this increased risk provides a basis for secondary intervention to reduce morbidity and mortality in diabetic patients. Hyperglycemia and protein glycation, increased inflammation, a prothrombotic state and endothelial dysfunction have all been implicated as possible mechanisms for such complications. A linking element between many of these phenomena could possibly be, among other factors, increased production of reactive oxygen species. Vascular endothelial cells have several physiological actions that are essential for the normal function of the cardiovascular system. These include the production of nitric oxide (NO), which regulates vasodilatation, anticoagulation, leukocyte adhesion, smooth muscle proliferation and the antioxidative capacity of endothelial cells. However, under conditions of hyperglycemia, excessive amounts of superoxide radicals are produced inside vascular cells and this can interfere with NO production leading to the possible complications. This article aims at reviewing the links between reactive oxygen species, diabetes and vascular disease and whether or not antioxidants can alter the course of vascular complications in diabetic patients and animal models. A possible beneficial effect of antioxidants might present a new addition to the range of secondary preventive measures used in diabetic patients.

In this study, Aspergillus terreus was irradiated by a 7.3 mW He-Ne laser in the presence of crystal violet, toluidine blue O and hematoporphyrin as photosensitizers. Xylanases recovered from non-irradiated and irradiated fungi were purified and characterized. The maximum production of xylanase (42.2 U/ml) was obtained after 5 min of laser irradiation in the absence of the photosensitizer. The irradiation of the sensitized fungus diminished the production of xylanase. On purification using G-100, the specific activity of xylanase recovered from the irradiated fungus was 292 U/mg protein representing a 37-fold purification over the crude extract compared with 95.6 U/mg protein representing the 12.8-fold for the enzyme recovered from the non-irradiated fungus. The enzyme recovered from the irradiated fungus had lower molecular weight as compared with that recovered from the non-irradiated one. Characterization of the purified enzymes revealed that the enzyme recovered from the irradiated fungus was more thermostable and had a wider range of optimum reaction temperature (60-70 degrees C) and pH (4.0-12.0), compared to the non-irradiated one.

In this study, Aspergillus terreus was irradiated by a 7.3 mW He-Ne laser in the presence of crystal violet, toluidine blue O and hematoporphyrin as photosensitizers. Xylanases recovered from non-irradiated and irradiated fungi were purified and characterized. The maximum production of xylanase (42.2 U/ml) was obtained after 5 min of laser irradiation in the absence of the photosensitizer. The irradiation of the sensitized fungus diminished the production of xylanase. On purification using G-100, the specific activity of xylanase recovered from the irradiated fungus was 292 U/mg protein representing a 37-fold purification over the crude extract compared with 95.6 U/mg protein representing the 12.8-fold for the enzyme recovered from the non-irradiated fungus. The enzyme recovered from the irradiated fungus had lower molecular weight as compared with that recovered from the non-irradiated one. Characterization of the purified enzymes revealed that the enzyme recovered from the irradiated fungus was more thermostable and had a wider range of optimum reaction temperature (60-70 degrees C) and pH (4.0-12.0), compared to the non-irradiated one.

BACKGROUND: The aim of this study was to determine if the number of treatment sessions has an influence on the final intermediate term results of submucosal bipolar radiofrequency volumetric tissue reduction of the inferior turbinate.

METHODS: One hundred two patients were enrolled in this study prospectively. The procedure was done using the Coblation (Arthrocare Corp., Sunnyvale, CA). The assessment was done using the 10-cm visual analog scale and acoustic rhinometry. Surgical procedures and pre- and postoperative assessments were done at the Faculty of Medicine, Cairo University.

RESULTS: Eighty eight percent of our study population achieved final relief of their nasal obstruction, and at least three sessions were needed to maintain the favorable outcome at 1-year follow up.

CONCLUSION: Increasing the number of bipolar radiofrequency volumetric tissue reduction treatment sessions was associated with better intermediate-term outcome. This was confirmed using subjective and objective methodology.

The homology and diversification of genomic sequence encoding glucagon gene among native Egyptian buffalos, camel and sheep were tested using cattle as model. Oligodeoxynucleotide primers designed from the available GenBank data were used for PCR probing of the glucagon gene encoding sequence at different loci. The DNA oligomer probes were constructed to flank either the whole gene encoding sequence or different intra-gene encoding sequences. The PCR products were visualized using agarose gel electrophoresis. All species showed a same size band of prepro-glucagon when PCR was used to amplify the whole gene encoding sequence. In contrary, amplifications of different intra-gene loci failed to give the same results. The results indicated variable degrees of diversity among old world ruminating ungulates in the glucagon gene encoding sequence. Compared with other ruminants, the variation appears predominantly in camel. Surprisingly, the similarity in size between both amplification products of whole gene encoding sequence and the proposed size of glucagon cDNA definitely excludes the possibility of large intervening introns spanning the genomic sequence of the glucagon gene in these species. This indicates that, in contrast to other tested mammals, the glucagon gene includes an essentially full-length copy of glucagon mRNA. The study revealed a possible new aspect of glucagon gene evolution in order to correlate its corresponding protein function among different ruminant species.

The homology and diversification of genomic sequence encoding glucagon gene among native Egyptian buffalos, camel and sheep were tested using cattle as model. Oligodeoxynucleotide primers designed from the available GenBank data were used for PCR probing of the glucagon gene encoding sequence at different loci. The DNA oligomer probes were constructed to flank either the whole gene encoding sequence or different intra-gene encoding sequences. The PCR products were visualized using agarose gel electrophoresis. All species showed a same size band of prepro-glucagon when PCR was used to amplify the whole gene encoding sequence. In contrary, amplifications of different intra-gene loci failed to give the same results. The results indicated variable degrees of diversity among old world ruminating ungulates in the glucagon gene encoding sequence. Compared with other ruminants, the variation appears predominantly in camel. Surprisingly, the similarity in size between both amplification products of whole gene encoding sequence and the proposed size of glucagon cDNA definitely excludes the possibility of large intervening introns spanning the genomic sequence of the glucagon gene in these species. This indicates that, in contrast to other tested mammals, the glucagon gene includes an essentially full-length copy of glucagon mRNA. The study revealed a possible new aspect of glucagon gene evolution in order to correlate its corresponding protein function among different ruminant species.

OBJECTIVES: To evaluate the clinical presentation and treatment end results of primary adenocarcinoma of the urinary bladder, and to determine the significant independent prognostic factors that determine this outcome.

PATIENTS AND METHODS: Of 3659 patients who underwent cystectomy, 192 had adenocarcinoma of the urinary bladder, with a relative frequency of 5.2%. Most of these patients (68.2%) presented in late stages (P3 + P4). The incidence of pelvic lymph nodes involvement was 25.5%. Mucinous adenocarcinoma was reported in 28 patients (14.6%), papillary in 20 (10.4%), signet ring in 14 (7.3%), while not otherwise specified was reported in 130 (67.7%) in the cystectomy specimens.

RESULTS: Mucinous and signet-ring histologic subtypes showed increased frequency of high stages and high grades, and more nodal involvement than the papillary and not otherwise specified. All patients were treated with radical cystectomy and pelvic lymphadenectomy with (69 patients) or without (123) postoperative radiotherapy. The 5-year disease-free survival rate was 46 +/- 4% for all patients with adenocarcinoma. Postoperative radiotherapy improved the disease-free survival significantly. The 5-year disease-free survival rate for the postoperative radiotherapy group was 61 +/- 6% compared to 37 +/- 5% for the cystectomy alone group (P = 0.002). Local control rate was significantly improved from 53 +/- 7% for cystectomy alone to 96 +/- 3% for postoperative radiotherapy patients (P = 0.00001). Distant metastases were the leading cause of death in the postoperative radiotherapy group.

CONCLUSIONS: Within the limitations provided by retrospective studies, it could be concluded that postoperative radiotherapy improved the disease-free survival through its effect on local control. The disease-free survival independent prognostic variables were tumor stage, postoperative radiotherapy, nodal involvement, and adenocarcinoma subclassification. These factors, except the adeno-subclassification, were also found to determine the local control rate. On the other hand, the independent prognostic factors for distant metastasis were lymph nodal involvement, stage, and adeno-subclassification.

INTRODUCTION: The technique most commonly used for penile lengthening is the release of the suspensory ligament in combination with an inverted V-Y skin plasty. This technique has drawbacks such as the possibility of reattachment of the penis to the pubis, a hump that forms at the base of the penis, in addition to alteration in the angle of erection.

AIM: In this work, we describe a new technique that overrides these drawbacks and minimize the loss of gained length.

METHODS: The suspensory ligament was released through a penopubic incision. The caudal flap of the resected ligaments was reflected caudally and sutured to the Buck's fascia. The V flap was incised. The caudal half of the V was deskinned, leaving a cranial skin-covered V flap, and a caudal, rectangular fat flap. The fat flap was pulled into the gap between the base of the penis and the pubis and secured in position by suturing its deep surface and lower edge to the pubis. This maneuver filled up the gap. The V incision was closed as a Y. The penopubic incision was closed as a T shape, to avoid pulling the penis back at skin closure. A stay suture stretched from the glans to the thigh, maintaining the penis in the stretched position. A urinary catheter was inserted.

RESULTS: Six months after surgery, there was no loss in the length gained. The angle of erection (as reported by the patient) was similar to that prior to the procedure. The skin incisions left no hump and a faint scar that was not troublesome to the patient.

CONCLUSION: "V-Y half-skin half-fat advancement flap" and "T-closure" may improve the results of suspensory ligament release for penile lengthening. The reported techniques minimize the losses compromising length gain, whether in-surgery or following it.