Rat nicastrin gene: cDNA isolation, mRNA variants and expression pattern analysis

Nicastrin is a type 1 transmembrane glycoprotein that interacts with presenilin, Aph-1, and Pen-2 proteins to form a high molecular complex with gamma secretase activity. Then, nicastrin has a central role in presenilin-mediated processing of beta-amyloid precursor protein and in some aspects of Notch/glp-1 signaling in vivo. Here, we isolated a rat nicastrin cDNA and investigated gene expression in embryonic and adult rat tissues. The predicted amino acid sequence is comprised of 708 residues and showed a high degree of identity with other vertebrate orthologs. Besides full-length nicastrin mRNA, we identified an alternative spliced variant lacking the whole exon 3 and predicted to encode a 62-residue-long truncated protein. Full-length nicastrin mRNA was observed to be ubiquitously expressed, while the spliced variant was preferentially transcribed in the nervous system, whether in embryonic or adult neural tissues. Studies performed on primary cell cultures demonstrated that the short isoform was expressed in neurons, but not in astrocyte and microglial cells. Further experiments performed to verify the presence of the variant in neuroblastoma culture failed to show any truncated protein. Treatments by cyclohexamide showed the involvement of a quality control-based surveillance mechanism, which selectively degrades the exon 3-skipped isoform. In summary, this is the first report describing a novel skipped isoform of nicastrin which may suggest a new possible control mechanism based on the alternative splicing and nonsense-mediated mRNA decay to regulate brain protein expression and provide newer insights into potential implication in Alzheimer's disease.     

Neonatal lupus erythematosus: 4 cases and clinical review

Neonatal lupus erythematosus (NLE) is an infrequent disease in newborns caused by the transplacental passage of maternal Anti-Ro/SSA, Anti-La/SSB and/or Anti-U1 RNP antibodies. The most common manifestations are cutaneous and cardiac. We carried out a retrospective study of cases of NLE diagnosed in the last 10 years at the Hospital Universitario Insular in Gran Canaria. Complete data was obtained for 4 patients. Three cases had circulating Anti-Ro antibodies in the mother and in the newborns, while in the fourth case they were Anti-RNP. Two mothers were diagnosed with systemic lupus, one with mixed connective tissue disease and the other with leucocytoclastic vasculitis. The skin lesions consisted of urticaria-like and desquamative lesions. One patient presented with ulceration. The histological study of the urticaria-like lesions showed a non-specific perivascular infiltrate; the desquamative lesions were consistent with subacute lupus erythematosus.     

Vascular but not cardiac remodeling is associated with superoxide production in angiotensin II hypertension

OBJECTIVE: Angiotensin (Ang) II increases reactive oxygen species (ROS), decreases nitric oxide (NO) bioavailability and promotes cardiovascular remodeling. ROS have been identified as critical second messengers of the trophic responses by Ang II. In rats with Ang II-induced hypertension, we investigated the role of ROS in cardiac hypertrophy as well as the remodeling of aortas and mesenteric (resistance) arteries. METHODS: Sprague-Dawley rats received Ang II (0.7 mg/kg per day by mini-pump, n = 7) or vehicle (n = 7) for 5 days. Endothelium-dependent relaxation to acetylcholine (EDR) in aortas was determined in organ baths and in mesenteric resistance vessels in a pressurized myograph. Superoxide (O2) production was measured by lucigenin chemiluminescence, laser-confocal fluorescence microscopy (LCM) and NADPH oxidase assay. RESULTS: Ang II-treated rats developed hypertension (183 +/- 3 versus 138 +/- 4 mmHg, P < 0.05), increased aortic O2 (50%), aortic hypertrophy (12%) and impaired EDR. Mesenteric arteries manifested impaired EDR, increased NADPH oxidase activity (356%) and eutrophic inward remodeling (decreased lumen diameter and increased wall/lumen ratio). However, although Ang II-treated rats developed cardiac hypertrophy (13%), this was not accompanied by an increase in cardiac O2, as measured by lucigenin, LCM or NADPH oxidase assay. On the other hand, cardiac calcineurin, a molecule that promotes cardiac hypertrophy linked to Ang II, was increased by 40% (52 +/- 8 versus 33 +/- 5 pmol/min per mg protein, P < 0.05). CONCLUSION: These studies demonstrate that the role of ROS in Ang II-induced vascular remodeling differ across vascular territories. Although in conduit and resistance vessels, vascular hypertrophy and endothelial dysfunction are linked to increased ROS production, cardiac hypertrophy is not. Instead, cardiac hypertrophy is associated, at least in part, with an increase in calcineurin. These studies unveil novel mechanisms that may play an important role in the pathogenesis of cardiac and vascular injury in hypertension.     

Preliminary experience with Leo self-expanding stent for the treatment of intracranial aneurysms

We report initial experience with the use of a new intracranial stent, the Leo microstent in combination with detachable coils in treatment of patients with wide-necked cerebral aneurysms. The Leo stent represents a significant advance in the vascular treatment of intracranial aneurysm with high radial force and an easy delivery system. It is a feasible, secure, and effective system.     

Robotic fundoplication for gastro-oesophageal reflux disease

Presented as a possible "second" revolution in general surgery after the introduction of laparoscopy during the last few years, the robotic approach to mini-invasive surgery has not yet witnessed wide, large-scale diffusion among general surgeons and is still considered an "experimental approach". In general surgery, the laparoscopic treatment of gastrooesophageal reflux is the second most frequently performed robot-assisted procedure after cholecystectomy. A review of the literature and an analysis of the costs may allow a preliminary evaluation of the pros and cons of robotic fundoplication, which may then be applicable to other general surgery procedures. Eleven articles report 91 cases of robotic fundoplication (75 Nissen, 9 Thal, 7 Toupet). To date, there is no evidence of benefit in terms of duration of surgery, rate of complications and hospital stay. Moreover, robotic fundoplication is more expensive than the traditional laparoscopic approach (the additional cost per procedure due to robotics is 1,882.97 euros). Only further technological upgrades and advances will make the use of robotics competitive in general surgery. The development of multi-functional instruments and of tactile feedback at the console, enlargement of the three-dimensional laparoscopic view and specific "team" training will enable the use of robotic surgery to be extended to increasingly difficult procedures and to non-specialised environments.     

Calcified renal artery aneurism and high blood pressure. A case report and review of the literature

Renal artery aneurysm is a rare disease in children and usually is due to fibromuscular dysplasia. Clinical symptoms are frequently high blood pressure, abdominal pain, and hematuria. Diagnosis is carried out by means of angiography. We report the case of a 13-year-old male patient who had renovascular hypertension due to calcification and aneurysm of fibromuscular dysplasia-associated renal artery. We carried out total nephrectomy to resolve high blood pressure. We suggested that presence of discreet calcification in region of renal artery in a boy with renovascular high blood pressure should guide us toward diagnosis of fibromuscular dysplasia-related renal artery aneurysm.     

The spreading of B lymphocytes induced by CD44 cross-linking requires actin, tubulin, and vimentin rearrangements

CD44 is a polymorphic family of adhesion molecules widely distributed on cells and tissues. CD44 is up-regulated on activated lymphocytes, and it can function as a receptor, mediating rolling and migration. Although it has been demonstrated that anti-CD44 antibodies bound to tissue-culture plates induce multidirectional emission of retractile dendrites ("spreading") in activated murine B lymphocytes, the involvement of cytoskeleton elements in this phenomenon is largely unknown. In this work, it is shown that the generation of dendrites induced by CD44 cross-linking in activated B cells depends on actin, microtubules, and vimentin reorganization. Immunofluorescence analysis showed that dendrite formation began with actin polymerization, and its extension was favored by microtubules and intermediate filaments of vimentin oriented to the polymerized actin. Pretreatment of activated B lymphocytes with cytochalasin E inhibited the dendrites formation; moreover, when cells were treated with this drug at different time points during the dendrite formation process, the stability of the dendrites was affected. In contrast, although the treatment with colchicine and nocodazole (tubulin polymerization inhibitors) inhibited the dendrites formation, it did not inhibit the initial phase of actin polymerization. According to these results, B cell spreading and dendrite formation induced by anti-CD44 antibodies require coordinated rearrangements of actin, microtubules, and vimentin, being the actin cytoskeleton, the most important element that confers stability and drives the morphological changes during B cell spreading, conceivably preparing B lymphocytes for locomotion.     

Structure-affinity relationship study on N-(1,2,3,4-tetrahydronaphthalen-1-yl)-4-aryl-1-piperazinealkylamides, a new class of 5-hydroxytryptamine7 receptor agents

A series of N-(1,2,3,4-tetrahydronaphthalen-1-yl)-4-aryl-1-piperazinealkylamides was prepared and their affinity for serotonin (5-hydroxytryptamine, 5-HT) 5-HT7, 5-HT(1A), and 5-HT(2A) receptors was measured by in vitro binding assays. In relation to 5-HT7 receptor affinity, receptor binding studies indicated that (i) the optimal alkyl chain length was five methylenes, (ii) an unsubstituted 1,2,3,4-tetrahydronaphthalenyl nucleus was preferred, and (iii) the substitution pattern of the aryl ring linked to the piperazine ring played a crucial role. Several compound with high affinity for 5-HT7 receptors were identified. Among them, 4-(2-methoxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (28), 4-(2-acetylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (34), 4-(2-methylthiophenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (44), 4-(2-hydroxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (46), and 4-(2-methylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (49) were assayed for the 5-HT7 receptor-mediated relaxation of substance P-induced guinea pig ileum contraction. Compounds 28, 44, and 49 behaved as full agonists and compound 34 as a partial agonist, whereas derivative 46 acted as an antagonist. Among the compounds presented here, it emerged that 44 was identified as a potent 5-HT7 receptor agonist (Ki = 0.22 nM, EC50 = 2.56 microM), endowed with selectivity over 5-HT(1A) and 5-HT(2A) receptors (200-fold and >1000-fold, respectively).     

Hyperfractionated radiotherapy for T2N0 glottic carcinoma: a retrospective analysis at 10 years follow-up in a series of 60 consecutive patients

AIMS AND BACKGROUND: To report results of hyperfractionated radiotherapy for T2N0 glottic carcinoma at a single institution after extended follow-up. METHODS: Between 1980 and 1988 at Varese University Hospital, 60 consecutive patients with T2N0 glottic carcinoma received exclusive radiotherapy consisting of 1.5 Gy/fraction twice a day. Treatment gaps during the radiotherapy course were allowed according to individual tolerance. This policy resulted in a wide range of elapsed treatment time: median, 5.7 weeks; range, 3.7-8.9. Median follow-up is 9.8 years. RESULTS: As a result of dose/time distribution, 16, 20 and 24 patients received an average weekly dose rate of <10 Gy/week, equal to 10 Gy/week or >10 Gy/week, respectively. Mean total dose for each group was 62.8 Gy, 63.7 Gy and 63.8 Gy, respectively. Five-year local-regional control was 69 +/- 6% (95% CI); ultimate local-regional control, including salvage surgery, was 78 +/- 5%. All failures were at the primary site, and no patient developed neck recurrence as first site of failure. The actuarial incidence of grade 2-3 late reactions at 5 years was 42 +/- 6%. Most late toxicity events were grade 2: only 2 patients developed grade 3 reactions and none grade 4. None of the several clinical and treatment-related variables showed any statistically significant impact on local-regional control or late toxicity at univariate and multivariate analysis. In particular, 3-year local-regional control rates were 73 +/- 11%, 84 +/- 8% and 69 +/- 10% for an average weekly dose rate of <10 Gy/week, equal to 10 Gy/week and >10 Gy/week, respectively (not significantly different). CONCLUSIONS: At a very long follow-up, the hyperfractionated regimen tested in the study was shown to be effective and devoid of major complications, provided individual patient acute tolerance is carefully taken into account. Also, time factor did not affect outcome in this series.