GC-rich coding sequences reduce transposon-like, small RNA-mediated transgene silencing.

The molecular basis of transgene susceptibility to silencing is poorly characterized in plants; thus, we evaluated several transgene design parameters as means to reduce heritable transgene silencing. Analyses of Arabidopsis plants with transgenes encoding a microalgal polyunsaturated fatty acid (PUFA) synthase revealed that small RNA (sRNA)-mediated silencing, combined with the use of repetitive regulatory elements, led to aggressive transposon-like silencing of canola-biased PUFA synthase transgenes. Diversifying regulatory sequences and using native microalgal coding sequences (CDSs) with higher GC content improved transgene expression and resulted in a remarkable trans-generational stability via reduced accumulation of sRNAs and DNA methylation. Further experiments in maize with transgenes individually expressing three crystal (Cry) proteins from Bacillus thuringiensis (Bt) tested the impact of CDS recoding using different codon bias tables. Transgenes with higher GC content exhibited increased transcript and protein accumulation. These results demonstrate that the sequence composition of transgene CDSs can directly impact silencing, providing design strategies for increasing transgene expression levels and reducing risks of heritable loss of transgene expression.

Canola engineered with a microalgal polyketide synthase-like system produces oil enriched in docosahexaenoic acid.

Dietary omega-3 long-chain polyunsaturated fatty acids (LC-PUFAs), docosahexaenoic acid (DHA, C22:6) and eicosapentaenoic acid (EPA, C20:5) are usually derived from marine fish. Although production of both EPA and DHA has been engineered into land plants, including Arabidopsis, Camelina sativa and Brassica juncea, neither has been produced in commercially relevant amounts in a widely grown crop. We report expression of a microalgal polyketide synthase-like PUFA synthase system, comprising three multidomain polypeptides and an accessory enzyme, in canola (Brassica napus) seeds. This transgenic enzyme system is expressed in the cytoplasm, and synthesizes DHA and EPA de novo from malonyl-CoA without substantially altering plastidial fatty acid production. Furthermore, there is no significant impact of DHA and EPA production on seed yield in either the greenhouse or the field. Canola oil processed from field-grown grain contains 3.7% DHA and 0.7% EPA, and can provide more than 600 mg of omega-3 LC-PUFAs in a 14 g serving.

Impact of gender differences on the outcome of Alzheimer's disease.

BACKGROUND AND AIMS: Since little is known about the role of gender in the course of Alzheimer's disease (AD), a prospective epidemiological study was conducted to detect gender differences in relation to AD evolution and outcome. METHODS: Six hundred AD patients, 214 men and 386 women, first seen between September 2000 and December 2003, were enrolled; the follow-up period lasted until December 2008. RESULTS: The men had greater comorbidity and higher mortality than the women, who instead recorded more disability and longer survival. Survival curves showed that women reach partial loss of autonomy faster than men. Higher Neuropsychiatric Inventory scores at baseline showed a predictive value for loss of autonomy regardless of gender. Pharmacological treatment seems to have a protective role on disability and mortality. CONCLUSIONS: Gender influences disease evolution not only directly but also through other factors such as comorbidity.

Different geometric patterns of pacifiers compared on the basis of finite element analysis.

AIM: This study was carried out with the purpose to show on a virtual model of oral cavity the mechanical behaviour of different kinds of pacifiers with different pressure levels that can be likened to a condition of rest and deglutition. MATERIALS AND METHODS: Three different types of dummies, orthodontic- (A), cherry- (B) and drop- (C) shaped from an anatomical point of view, were inserted between the palate and the tongue in a virtual system by means of a finite element simulation. The palatal structure was recreated through tridimensional laser scanning, while the tongue structure was reconstructed by a software suitable for reproducing solids. Also the image of the pacifiers was developed by computer-aided scanning and reproduction. Suitable constraints were inserted and high and low pressure levels were exerted on these systems. FEA simulation allowed us to distribute the strain on the palate according to the different geometrical structures of the objects. RESULTS: Dummy A shows a more uniform and wider crosswise stress distribution with also a lesser load on the anterior palatal crest. Dummy B and C, on the contrary, show a more dot-like behaviour inducing a higher stress due to contact on restricted points. CONCLUSION: The characteristics of dummy A, although they have not been clinically investigated yet, seem to be the fittest ones to guarantee the maintenance of the transversal diameters of the premaxilla and reduce the risk of open bite.

DNp73alpha protects myogenic cells from apoptosis.

The P73 gene is transcribed from two promoters, P1 and P2, that direct the expression of multiple transactivation competent (TA) and dominant negative (DN) isoforms. TAp73 transcription factors mediate cell cycle arrest and/or apoptosis in response to DNA damage and are involved in developmental processes. P73 mRNA levels increase and the P1p73 promoter is upregulated during myogenic differentiation of C2C12 skeletal muscle satellite cells. The DNp73 proteins act as trans-repressors of p53- and p73-dependent transcription, and possess both antiapoptotic and pro-proliferative potential. Here, we show that DNp73alpha is expressed in proliferating C2C12 myoblasts, rapidly accumulates in differentiating myocytes and remains elevated in C2C12 myotubes. By combining transactivation assays and chromatin immunoprecipitation analysis, we could show that the upregulation of the P2p73 promoter during myogenic differentiation is mediated by the coordinated recruitment and activity of MyoD and p53/p73. Abrogation of DNp73 expression by specific siRNA led to a strong potentiation of the spontaneous apoptosis of C2C12 myoblasts induced to differentiate. Finally, unlike TAp73 that contributes to DNA damage-induced apoptosis of myotubes, endogenous DNp73 mediates the relative resistance of differentiated myotubes to DNA damage. Altogether, our findings identify DNp73alpha as an important target in designing strategies aimed at the potentiation of the regenerative potential of skeletal satellite cells.

Spontaneous intracranial hypotension: case report.

Spontaneous intracranial hypotension is a well defined entity, however it is characterised by a relevant clinical heterogeneity. The main feature is orthostatic headache, often accompanied by other symptoms. The diagnosis is easily made in the presence of orthostatic headache, excluding an obvious aetiology of low cerebrospinal fluid (CSF) pressure, and often there is no evidence of direct or indirect CSF leak.

[++Scintimammography: a new imaging technique for diagnosis and follow-up of breast cancer]. / Apport de la mammoscintigraphie 99m Tc MiBi dans le diagnostic et le suivi des cancers du sein.

99m Tc MiBi has emerged as a new imaging technique for primary breast carcinomas. The aim of this study was to analyze the diagnostic performance and the additional information provided by scintimammography (SMN). Two hundred and forty consecutive women referred to surgery for abnormalities breast or suspicious breast cancer were scanned before surgery. Sensitivity and specificity were respectively 92% and 53%. PPV: 77.5% and PVN: 79.3%. All false negative scans occurred in cancers 1 cm. SMN detected multiple foci of uptake in the same breast in 13%, that were all confirmed to be multifocal disease and histology. Controlateral focal uptake was also detected; at this time 5/27 are confirmed to be bilateral breast neoplasms; 22 patients are in follow-up. SMN is reliable in the diagnosis of breast cancer and also with difficult cases of mammography. Moreover, SMN provides additional qualitative information in 19.6% of breast carcinomas, such as chest wall infiltration, multifocal or bilateral breast cancer.

Long-Term Nicergoline Treatment of Mild to Moderate Senile Dementia : Results of a Multicentre, Double-Blind, Placebo-Controlled Study.

The efficacy and tolerability of nicergoline were evaluated in a long-term, double-blind, placebo-controlled trial. 108 patients, fulfilling DSM III-R criteria for mild to moderate senile dementia of degenerative, vascular or mixed origin, were selected from a pool of outpatients attending five Italian neurological centres and randomised to receive nicergoline 30mg twice daily (54 patients) or placebo (54 patients) for 12 months. Treatment efficacy on cognitive and behavioural performances was assessed by the Sandoz Clinical Assessment Geriatric scale (SCAG) and Mini Mental State Examination (MMSE), at baseline and after 3, 6, 9 and 12 months of treatment. Investigators and patients or caregivers provided a global evaluation of treatment outcome at study end. The efficacy analysis was carried out on 101 patients (51 nicergoline; 50 placebo) who completed the 12-month study. At study end, the SCAG total score and its clusters showed statistically significant improvement in the nicergoline-treated group compared with placebo-treated patients. The MMSE total score was maintained with nicergoline treatment. Global treatment evaluations, both by physician and patients, were consistently in favour of nicergoline (p < 0.001). Nicergoline was well tolerated; incidence of adverse events (7% in the nicergoline and 2% in the placebo group), withdrawals and haemodynamic changes were comparable with placebo.

Spinal chondroma of the lumbar tract: case report.

BACKGROUND: Cartilage-forming tumors are benign cartilaginous tumors that rarely affect the spinal canal: they account for 2% of all spinal tumors and 2.6% of all benign bone tumors. Pathologically, they may be classified as chondromas, osteochondromas, chondroblastomas, and chondromyxoid fibromas. This oncotype may remain asymptomatic (it is confined within the vertebral structure) or may present as a hard paravertebral swelling (it invades the paravertebral structures) or more rarely, with a slowly-developing neurologic syndrome (it extends into the vertebral canal). METHODS: Thirty-one cases have been reported (including our case) of benign cartilage-forming tumors localized in the lumbar column. Only three cases of chondroma of the lumbar spine presented with lumbar radicular pain. We report a fourth case and review clinical and radiologic characteristics of these lesions. RESULTS: Eleven out of the 31 cases were diagnosed as chondromas, 17 as osteochondromas, while in three cases the histopathologic diagnosis was not reported. Seventeen cases originated from the neural arch, seven from the vertebral body, two from the spinous process, and in five cases the exact localization was not reported. This tumor is more frequent in males (21 cases out of 31), than in females (five cases); in five cases the sex was not reported. Mean duration of symptoms was 23 +/- 5.1 months (range: 1-96); chondromas have a short clinical history before diagnosis (13.8 +/- 3.4 months) compared to osteochondromas (28.6 +/- 7.6). Clinical presentation with local swelling is reported in 10 cases, in 10 cases local pain without radicular irradiation, in six cases lumbar pain with sciatica, in two cases signs and symptoms of cord compression, one case of cauda syndrome, while in four cases no clinical details are reported. Among the six cases presenting with sciatica, four were chondromas (in all cases the L4 level was involved), and one osteochondroma, while in one case the histopathologic diagnosis was not reported. CONCLUSION: Computed tomography is important and indispensable for preoperative diagnosis, giving a precise indication of tumor extent and location and its relationship to the adjacent structures; while MRI is helpful in detecting patterns related to histologic malignancy. It is important to examine the whole tumor histologically because it is known that there may be small areas that show signs of malignancy; thus is more likely in chondromas than osteochondromas.

An assessment of hypothalamo-pituitary-adrenal axis functioning in non-depressed, early abstinent alcoholics.

Chronic alcohol consumption has been shown to be associated with abnormalities in the regulation of the hypothalamo-pituitary-adrenal (HPA) axis in humans. However, conflicting data exist in the literature, with particular regard to studies performed in actively drinking or withdrawn alcoholics; in addition, the frequent presence of depressive disturbances in such patients may importantly affect the neuroendocrine findings. In this study, we investigated HPA function in 12 male alcoholics, in whom the presence of depression and other possible confounding factors was excluded, during the first and second weeks after cessation of ethanol intake. The plasma corticotropin (adrenocorticotropic hormone, ACTH) and cortisol levels in response to both a stimulation test with human corticotrophin-releasing hormone (CRH; 100 micrograms IV) and an insulin (0.15 UI/kg IV)-induced hypoglycaemia (ITT) were measured; the cortisol response to a standard overnight dexamethasone (1 mg) suppression test (DST) was also tested. While the mean baseline ACTH and cortisol levels, measured in the morning (0800-0830 h), were not different from those of controls, ACTH and cortisol responses to the CRH test were markedly reduced (area of secretion p < .01 and p < .05, compared to controls). Similarly, the patient group showed an almost absent ACTH and cortisol release following insulin infusion (area of secretion p < .01 compared to controls, in either case). In four patients, non-suppression of plasma cortisol levels was seen on the DST, but no significant difference from normal suppressors was noted as far as the clinical features were concerned. These findings suggest that impaired hypothalamic and pituitary responsiveness, unrelated to depressive disturbances, occurs in recently withdrawn chronic alcoholics. While the possible influence of the alcohol withdrawal syndrome should be taken into account, such a pattern may be due to increased activity of the HPA axis, even in the face of preserved basal adrenal secretion. Whether these findings reflect a direct effect of sustained ethanol exposure on the components of the HPA axis, or a non-specific marker of impaired adaptation in chronic alcoholics, deserves further investigation.

Symptomatic headaches.

This paper is aimed to discuss some aspects of so-called symptomatic or secondary headaches with particular reference to the diagnostic criteria proposed by the IHS Classification Committee on headache disorders, cranial neuralgias and other head pain. Symptomatic headaches, in fact, represent an heterogenous group of conditions, sometimes with relevant clinical importance as warning symptoms of major intracranial pathology, sometimes benign in nature and with ill-defined clinical pictures and unclear relationship with the causative factor or event. It is therefore important to establish general principles for a more systematic approach to the problem of this group of headaches, also considering that some particular forms of secondary headaches, may be viewed as reliable "in-vivo" models for pathophysiological investigation in head pain.

[Not Available]. / Trattamento con talidomide delle ulcere oro-faringo-esofagee in AIDS: descrizione di un caso.

We report a case of oro-pharyngeal and oesophageal ulcerative disease in a AIDS patient. The patient complained of severe oral pain and dysphagia. Systemic manifestations as fever and fatigue were also present. Repeated microbiological exams were negative and therapy with either antibiotics or antiviral agents or corticosteroids or colchicine was unsuccessful. Thus, we started thalidomide: 200 mg/day X 21 days, followed by 100 mg/day X 25 days. A dramatic reduction of symptoms was observed within 15 days from the onset of treatment. The healing of ulcers was evident already one month after the start of therapy. No relevant side effects were reported. Among those patients with AIDS at low risk of side effects, thalidomide may represent a successful therapeutic presidium for severe ulcerative disease of the gastrointestinal tract.

Pramiracetam effects on scopolamine-induced amnesia in healthy volunteers.

Pramiracetam has been evaluated for its potential antiamnesic properties in scopolamine-induced amnesia in healthy volunteers. Two groups of twelve males, 18-42 and 55-65 years old, respectively, were randomly assigned to oral treatment with pramiracetam (600 mg twice a day) or with placebo for 10 consecutive days. On day 11 each subject was injected intramuscularly with scopolamine hydrobromide (0.5 mg). Before scopolamine injection and then 1, 3 and 6 h after it, subjects were administered the following psychometric tests: simple and choice visual reaction times, digit symbol substitution test, Rey's 15 words test for short and long term verbal memory. Scopolamine significantly impaired episodic memory and selective attention tests in both scopolamine and placebo groups. Instead visuo-motor and incidental learning measures were unaffected. Pramiracetam, when compared to placebo, was able to partially reduce the amnesic effects induced by scopolamine both in young and old subjects.

Characterization of a genomic sequence coding for potato multicystatin, an eight-domain cysteine proteinase inhibitor.

A gene coding for potato multicystatin (PMC), the crystalline inhibitor of cysteine proteases which is found in tubers, was isolated and characterized. The deduced polypeptide product of this genomic sequence is 757 amino acids long and has a molecular mass of 86,778 Da. It consists exclusively of eight closely related domains, with 53-89% identity of residues. Each repeated unit is homologous to the cystatin superfamily of cysteine protease inhibitors. To date, no other member of this family has been found to contain so many inhibitor domains in one polypeptide. Eight introns are proposed in the 3.5 kb of genomic DNA coding for PMC, one in each cystatin unit. There is a family of 4 to 6 such large genes in potato, while in pea and maize the homologues are much smaller, and probably code for single-domain cystatins. PMC transcripts are abundant in tubers, but scarce in undamaged leaves or stems of field-grown potatoes. The tuber messages are derived from at least four genes (including the cloned example). The pattern of gene expression, as well as the properties of the protein, suggest that PMC has a role in the plant's defense system.

[Can benign and malignant thyroid nodules be differentiated with thallium 201?]. / Le thallium 201 permet-il de différencier le nodule thyroïdien bénin du nodule malin?

The merits of Thallium 201 radionuclide scanning of the thyroid mentioned as soon as 1976 by PALERMO have been confirmed in the 16 last studies published in the international literature. Over 1601 examinations, authors showed that any cold nodule (Tc 99 m or I 123) which preferentially fixes Thallium 201 as compared to surround thyroid tissue must be operated since some of them are thyroid cancer carriers, some others are carriers of a traditional follicular adenoma or Hurthle-cell adenoma, or a follicular adenoma associated with varied cell atypisms which make difficult the diagnosis between benign and malignant nodules. The analysis of the results published show a 91.3% sensitivity of this diagnosis test, all analysis methods being considered. In case of negative test, it allows eliminating the cancer risk by more than 97% (negative predictive value). Such an examination--as opposed to a cytology test on the isolated nodule only--is valid for any type of nodule, being isolated, multiple or a multinodular goitre where the degeneration risk is close to, or even higher than, that of isolated nodules (4, 22).

[Economic analysis of diagnostic and therapeutic methods in cases of cold thyroid nodules]. / Analyse économique des approaches diagnostiques et thérapeutiques du nodule froid de la thyroïde.

This article studies how two diagnostic tests (cytological puncture and 201thallium scanning) can be combined to obtain a more efficient approach of cold thyroid nodules. Diagnostic and therapeutic approaches are compared on the basis of four criteria: financial cost (immediate and delayed), number of labelled cancers, pre- and postoperative death rates and number of complications.

Correlation of 99Tcm-sestamibi SPECT with coronary angiography in general hospital practice.

One hundred and twenty-eight patients had a stress test with sestamibi injection and a coronarography within 2 months of the sestamibi study. Sestamibi studies were reconstructed using Hanning 0.65 and Wiener 1.0 filters; the latter improved the results significantly (P < 0.05). Significant correlations were found between sestamibi and coronarography: the sensitivity was 0.967 and the specificity was 0.711. Thus sestamibi is a very good compound, with excellent imaging quality, which is very sensitive for mild defects, even more sensitive than coronarography. Thorough studies of discrepancies show that they could be explained by reductions in coronary flow or myocardial reserve, even when the large epicardial vessels are disease free as occurs in various coronary involvements. A follow-up of positive patients with sestamibi and negative coronarography is absolutely necessary since this group of patients represents a 'risk' group. Moreover, sestamibi single photon emission computed tomographic accuracy is highly dependent on filter choice.

Long-term idebenone treatment of vascular and degenerative brain disorders of the elderly.

A multicentre study with an open experimental design was carried out on 118 patients suffering from mild to moderate cognitive decline due to cerebrovascular and degenerative disorders (chronic cerebrovascular disorders, CCVD; multi-infarct dementia, MID; aging brain, AB; dementia of Alzheimer's type, DAT). All patients, after a wash-out period of 3 weeks, were treated with idebenone (45 mg twice daily by oral route) for a period of 6 months. Behavioral and cognitive measures (Sandoz Clinical Assessment of Geriatrics, SCAG; Serial Learning Test) were applied to evaluate the long term therapeutical aspects. The results, analyzed by multivariate analysis of variance and chi2 test, showed a significant improvement of the cognitive profile in all patients, more evident in CCVD and AB groups. No remarkable side-effects were found in all groups of patients, thus confirming good tolerability of idebenone.