The microRNA-29a Modulates Serotonin 5-HT7 Receptor Expression and Its Effects on Hippocampal Neuronal Morphology.

miRNAs are master regulators of gene expression in diverse biological processes, including the modulation of neuronal cytoarchitecture. The identification of their physiological target genes remains one of the outstanding challenges. Recently, it has been demonstrated that the activation of serotonin receptor 7 (5-HT7R) plays a key role in regulating the neuronal structure, synaptogenesis, and synaptic plasticity during embryonic and early postnatal development of the central nervous system (CNS). In order to identify putative miRNAs targeting the 3'UTR of 5-HT7R mouse transcript, we used a computational prediction tool and detected the miR-29 family members as the only candidates. Thus, since miR-29a is more expressed than other members in the brain, we investigated its possible involvement in the regulation of neuronal morphology mediated by 5-HT7R. By luciferase assay, we show that miR-29a can act as a post-transcriptional regulator of 5-HT7R mRNA. Indeed, it downregulates 5-HT7R gene expression in cultured hippocampal neurons, while the expression of other serotonin receptors is not affected. From a functional point of view, miR-29a overexpression in hippocampal primary cultures impairs the 5HT7R-dependent neurite elongation and remodeling through the inhibition of the ERK intracellular signaling pathway. In vivo, the upregulation of miR-29a in the developing hippocampus parallels with the downregulation of 5-HT7R expression, supporting the hypothesis that this miRNA is a physiological modulator of 5-HT7R expression in the CNS.

Epithelioid angiosarcoma arising in schwannoma of the kidney: report of the first case and review of the literature.

BACKGROUND: Schwannoma and angiosarcoma are infrequent pathologies that have been rarely reported in the kidney. Angiosarcoma is an uncommon malignant tumor presenting a recognizable vascular differentiation. It can develop in any site but the most common locations include the skin, soft tissues, breast, bone, liver, and spleen while renal localization has been very rarely reported in the literature. Schwannoma is a benign peripheral nerve sheath tumor composed of cells with the immunophenotype and ultrastructural features of differentiated Schwann cells. It has a wide anatomical distribution but the most frequent locations include subcutaneous tissues of the extremities and the head and neck region and the retroperitoneal and mediastinal soft tissues. The occurrence of an angiosarcoma in a pre-existing schwannoma is an extremely rare event with <20 cases reported in worldwide literature. In the present study, a renal case of angiosarcoma arising in schwannoma is presented with a detailed review of the pertinent literature. CASE PRESENTATION: A 56-year-old man was admitted with a few days history of lower back pain and hematuria. Abdominal ultrasound showed a mass inside the left renal medulla. Subsequent imaging investigations with computed tomography and magnetic resonance confirmed the presence of the lesion and showed a pulmonary metastasis. CONCLUSIONS: The final histopathological examination led to the diagnosis of epithelioid angiosarcoma arising in a schwannoma. The patient came to death a few months later due to a massive hemothorax. To the best of our knowledge, the present is the first case of an angiosarcoma arising in a schwannoma of the kidney.

The serotonin receptor 7 promotes neurite outgrowth via ERK and Cdk5 signaling pathways.

Serotonergic neurotransmission is mediated by at least 14 subtypes of 5-HT receptors. Among these, the CNS serotonin receptor 7 (5-HTR7) is involved in diverse physiological processes. Here we show that treatment of murine striatal and cortical neuronal cultures with 5-HTR7 agonists (8-OH-DPAT and LP-211) significantly enhances neurite outgrowth. This effect is abolished by the selective 5-HTR7 antagonist SB-269970, by the ERK inhibitor U0126, by the cyclin-dependent kinase 5 (Cdk5) inhibitor roscovitine, as well as by cycloheximide, an inhibitor of protein synthesis. These data indicate that 5-HTR7 activation stimulates extensive neurite elongation in CNS primary cultures, subserved by ERK and Cdk5 activation, and de novo protein synthesis. Two-dimensional (2D) gel electrophoresis coupled to Western blot analyses reveals both qualitative and quantitative expression changes in selected cytoskeletal proteins, following treatment of striatal primary cultures with LP-211. In particular, the 34 kDa isoform of MAP1B is selectively expressed in stimulated cultures, consistent with a role of this protein in tubulin polymerization and neurite elongation. In summary, our results show that agonist-dependent activation of the endogenous 5-HTR7 in CNS neuronal primary cultures stimulates ERK- and Cdk5-dependent neurite outgrowth, sustained by modifications of cytoskeletal proteins. These data support the hypothesis that the 5-HTR7 might play a crucial role in shaping neuronal morphology and behaviorally relevant neuronal networks, paving the way to new approaches able to modulate CNS connectivity.

[Microbiological verification of a self control plan for a hospital food service]. / Verifica microbiologica di un piano di autocontrollo in un servizio di ristorazione ospedaliera.

During the past years, it has been an increment of food related infectious diseases. In order to avoid micro biological food contamination, adherence to good manufacturing is required through control measures of food safety practices. Updated national and European regulations underline the need to apply the HACCP system, overcoming the old concept of sample control on the end user product. This work shows results of microbiological controls made along the whole productive chain. Measurements are made using biomolecular techniques (PFGE) in order to assess the management of the micro biological risk of the self control plan applied to a hospital food service of Naples. The use of the PFGE applied on some micro-organisms gram negative potentially pathogen, underlines the circulation, continued in time, of these micro-organisms within the cooking area. In addition, cross contamination between several matrixes of samples has been detected.

Clinical and environmental distribution of Legionella pneumophila in a university hospital in Italy: efficacy of ultraviolet disinfection.

The molecular epidemiology of Legionella pneumophila in the 'V. Monaldi' University Hospital was studied. Seven cases of nosocomial Legionnaires' disease were diagnosed between 1999 and 2003. Two clinical legionella strains obtained from two patients in the adult cardiac surgery unit (CSU) and 30 environmental legionella strains from the paediatric and adult CSUs, neonatal intensive care unit (NICU) and the cardiorespiratory intensive care unit (CR-ICU) were serotyped and genotyped. L. pneumophila serogroup 1/Philadelphia with an identical pulsed-field gel electrophoresis (PFGE) profile A was isolated from two patients in the adult CSU, and from three and one water samples taken in the adult CSU and the paediatric CSU, respectively, from 2001 to 2002. Furthermore, L. pneumophila serogroup 3 with an identical PFGE profile B was identified in 20 environmental strains from all wards, L. pneumophila serogroup 3 with PFGE profile C was identified in a single environmental strain from the CR-ICU, and non-pneumophila Legionella with identical PFGE profile D was identified in five environmental strains from the adult CSU, paediatric CSU and NICU. Ultraviolet irradiation was effective in disinfection of the hospital water supplies in the adult and paediatric CSUs contaminated by L. pneumophila clone associated with nosocomial Legionnaires' disease. In conclusion, these data demonstrate that two cases of nosocomial legionellosis were caused by the persistence of a single clone of L. pneumophila serogroup 1/Philadelphia in the hospital environment, and that disinfection by ultraviolet irradiation may represent an effective measure to prevent nosocomial Legionnaires' disease.

Spinal intradural extramedullary haemangioma: MRI and neurosurgical findings.

Haemangiomas, have rarely been encountered in the spinal intradural extramedullary space and the MRI findings of this entity have been described only in a few cases. We present the Magnetic Resonance Imaging (MRI) and surgical findings of a rare case of intradural extramedullary cavernous angioma located at the T1-T2 level in a 65-year-old man presented progressive paraparesis and upper thoracic back pain. On MRI, a well-circumscribed intradural solid mass, 1 cm in diameter, was detected and another enhancing nodular mass was found at the nerve roots of the cauda equina. The thoracic spinal lesion was removed and the histological diagnosis confirmed cavernous haemangioma. Although very uncommon, haemangioma should be included in the differential diagnosis when a spinal intradural extramedullary lesion is discovered and some neuroradiological findings could allow a presumptive diagnosis.

[Cluster of Pseudomonas aeruginosa in a neurosurgical unit]. / Focolaio epidemico da Pseudomonas aeruginosa in un reparto di neurochirurgia.

Hospital infections caused by Pseudomonas aeruginosa and particulary meningitis appear to be very common in neurosurgical wards and in patients with brain traumas. This study was carried out during the period between 25th October and the 11th December 2000 in a neurosurgical ward of the teaching hospital of the University "Federico II" of Naples. During this period, five patients contracted an infection caused by P. aeruginosa. The microorganisim was found to be responsible for three cases of meningitis and two cases of surgical site infections. The P. aeruginosa isolates responsible for the infections and the "environmental" isolates were subjected to genotypic typing through the analysis of macrorestriction patterns of genomic DNA after XbaI digestion of Pulsed Field Gel Electrophoresis (PFGE). Four clinical isolates and an environmental isolate recovered from an hand washing basin located in the sub-intensive care area showed an identical PFGE pattern, as well as the some multiresistant antibiotype. The results of this study allows us to point out that the surveillance programs of infections in hospitals should include the molecular typing of micro-organisms singled out in clinical samples and, in case of outbreaks, also the typing of microorganisms originating from the environment and the hospital staff.

A nosocomial outbreak of Serratia marcescens producing inducible Amp C-type beta-lactamase enzyme and carrying antimicrobial resistance genes within a class 1 integron.

We investigated an outbreak of Serratia marcescens in the adult intensive care unit of the University Hospital of Napoli. The outbreak involved 13 cases of infection by S. marcescens over a nine-month period and was caused by a single pulsed-field gel electrophoresis clone. The epidemic strain was multiply antibiotic resistant, producing an inducible Amp C-type beta-lactamase enzyme and carrying the trimethoprim-resistance gene and the adenyltransferase gene, which confers resistance to streptomycin and spectinomycin, within a class 1 integron. Antimicrobial therapy with beta-lactams was associated with S. marcescens acquisition in the intensive care unit.

Molecular typing of Aeromonas isolates in natural mineral waters.

A total of 103 isolates of Aeromonas spp. were obtained over a 3-year period from a natural mineral water and from surface streams located within the boundaries of the watershed of the natural mineral water wells and were typed by macrorestriction analysis of genomic DNA with XbaI and by pulsed-field gel electrophoresis. All Aeromonas caviae isolates from the natural mineral water belonged to the same clone, and an analogous clonal identity was found among Aeromonas hydrophila isolates. These two clones expressed no hemolytic or cytotoxic activities. Aeromonas isolates from surface waters showed high molecular heterogeneity and were not related to the clones found in the natural mineral water. The presence of aeromonads chronically found in the natural mineral water was a likely consequence of a localized development of a biofilm, with no exogenous contamination of the aquifer. Molecular fingerprinting of drinking water isolates is a useful tool in explaining possible reasons for bacterial occurrences.

[Genomic typing techniques in a case of skin infection epidemic by Staphylococcus aureus in a newborn unit]. / Utilizzo delle tecniche di tipizzazione genotipica in un focolaio epidemico da Stafilococco aureo in un reparto di neonatologia.

In the present study it has been attempted to characterize the source, the reservoir and the modality of transmission of a skin infection epidemic by Staphylococcus aureus in a Newborn Unit. Every S. aureus isolate from clinician and environmental samples has been submitted to phenotypic typing, by means of antibiotic resistance patterns. The genotype has been analyzed using genomic DNA profile of restriction with Sma I and Pulsed Field Gel Electrophoresis (PFGE). In bacterial infections the epidemiologist nexus can think demonstrated when the stocks isolate from various cases belong to the same one clone. In the Newborn Unit the same one clone of S. aureus has been found on 2 health care workers, on 13 staff bearers characterized, and skin of 5 ill newborn infants. Therefore it has been able to go back to the probable source of infection and to take the right measures.

Molecular epidemiology of Stenotrophomonas maltophilia in a university hospital.

The aim of this investigation was to study the molecular epidemiology of Stenotrophomonas maltophilia in a university hospital in Italy. Sixty-one clinical isolates were collected from 43 patients during a two-year period. The majority of specimens were from the respiratory tract (41 of 43) of patients in the adult intensive care unit (ICU) (19 of 43) or cystic fibrosis (CF) patients (13 of 43). Genotypic analysis by pulsed-field gel electrophoresis (PFGE) of clinical isolates identified 31 different PFGE patterns. Although most patients were infected or colonized by different S. maltophilia clones, clones with identical genotype were isolated in patients from ICU, where two separate outbreaks were identified. Antimicrobial susceptibility identified a multi-resistant phenotype in all S. maltophilia PFGE clones. The majority of PFGE clones identified (six of seven clones from patients in the ICU) were susceptible to fluoroquinolones. Mechanical ventilation was associated with S. maltophilia acquisition in the ICU.

Messenger RNAs in synaptosomal fractions from rat brain.

Synaptosomal fractions from rat brain have been analyzed with semi-quantitative RT-PCR methods to determine their content of mRNAs coding for presynaptic, postsynaptic, glial, and neuronal proteins. Each mRNA was determined with reference to the standard HPRT mRNA. In our analyses, mRNAs were considered to be associated with synaptosomes only if their relative amounts were higher than in microsomes prepared in a polysome stabilizing medium, rich in Mg(++) and K(+) ions, or in the homogenate. According to this stringent criterion, the following synaptosomal mRNAs could not be attributed to microsomal contamination and were assumed to derive from the subcellular structures known to harbor their translation products, i.e. GAT-1 mRNAs from presynaptic terminals and glial processes, MAP2 mRNA from dendrites, GFAP mRNA from glial processes, and TAU mRNA from neuronal fragments. This interpretation is in agreement with the involvement of extrasomatic mRNAs in local translation processes.

Molecular epidemiology of an outbreak of Serratia marcescens in a neonatal intensive care unit.

OBJECTIVE: To investigate and control a biphasic outbreak of Serratia marcescens in a neonatal intensive care unit (NICU). DESIGN: Epidemiological and laboratory investigation of the outbreak. SETTING: The NICU of the 1,470-bed teaching hospital of the University "Federico II," Naples, Italy. PATIENTS: The outbreak involved 56 cases of colonization by S marcescens over a 15-month period, with two epidemic peaks of 6 and 3 months, respectively. Fourteen (25%) of the 56 colonized infants developed clinical infections, 50% of which were major (sepsis, meningitis, or pneumonia). METHODS: Epidemiological and microbiological investigations, analysis of macrorestriction pattern of genomic DNA through pulsed-field gel electrophoresis (PFGE) of clinical and environmental isolates, and institution of infection control measures. RESULTS: Analysis of macrorestriction patterns of genomic DNA by PFGE demonstrated that the vast majority of S marcescens isolates, including three environmental strains isolated from two handwashing disinfectants and the hands of a nurse, were of the same clonal type. The successful control of the outbreak was achieved through cohorting of noncolonized infants, isolation of S marcescens-infected and -colonized infants, and an intense educational program that emphasized the need for adherence to glove use and handwashing policies. The NICU remained open to new admissions. CONCLUSIONS: Outbreaks caused by S marcescens are very difficult to eradicate. An infection control program that includes molecular typing of microorganisms and the proper dissemination among staff members of the typing results is likely to be very effective in reducing NICU-acquired infections and in controlling outbreaks caused by S marcescens, as well as other multiresistant bacteria.

Diagnosis and therapy of intrathecal bleeding.

AIM: Define pathophysiology, epidemiology, diagnosis and therapy in case of spinal bleeding after central neural blockade (CNB). METHODS: Spinal epidural hematoma (SEH) following CNB may occur due to vascular trauma from needle/catheter placement and can occur in subdural and epidural spaces. Epidural artery bleeding seems the source of SEH: the damage mechanism depends on compression and neural vascular ischemia of cord, nerve roots, ganglion and toxicity from blood cell lysis products. Incidence varies (1:150.000 - 1:500.000) but SEH may be asymptomatic. CONCLUSIONS: SEH starts with acute severe low back and/or radicular pain and neurologic signs that may progress to paraparesis, sensory loss and sphincter disturbances. After CNB, the only sign of SEH may be an unusually prolonged motor and sensory block. Symptoms may start even 96 hours after CNB and/or removal of the epidural catheter. Neurological recovery is related to severity and speed of preoperative deficits development and surgical decompression. MR imaging features (diagnostic tool of choice), including degree of cord compression, are useful to establish or confirm the diagnosis of SEH but do not influence the management or predict outcome. Hematoma resolution and severity of neurologic impairment has the greatest impact on management and outcome. Preoperative MRI information and intraoperative evidence of subarachnoid hemorrhage (SAH) and CSF leakage is important: SAH worsens outcome for its negative effect on spinal cord and cauda equina. Conservative therapy may be successful in cases with minimal neurologic deficits, despite cord compression.

rTLE3, a newly identified transducin-like enhancer of split, is induced by depolarization in brain.

The transducin-like enhancers of split are a family of mammalian proteins that share sequence homology with the Drosophila protein Groucho. Using representational difference analysis, we isolated the cDNA for a previously unidentified gene, rTLE3 (rat transducin-like enhancer of split 3), as a sequence induced by depolarization and forskolin, but not by neurotrophins or growth factors, in PC12 pheochromocytoma cells. rTLE3 encodes the protein rTLE3, a 764-amino acid orthologue of mouse and human TLE3. R-esp2, the gene encoding the closest related rat protein, is not induced by any of the four treatments in PC12 cells. rTLE3 and R-esp2 have different patterns of expression in the adult rat CNS and other tissues. After systemic administration of kainic acid, rTLE3 is induced specifically in the dentate gyrus of the hippocampus. We propose that members of the transducin-like enhancer of split family of proteins may have distinct functions in the mature CNS, in addition to their functions during development.

The salt-inducible kinase, SIK, is induced by depolarization in brain.

Membrane depolarization of neurons is thought to lead to changes in gene expression that modulate neuronal plasticity. We used representational difference analysis to identify a group of cDNAs that are induced by membrane depolarization or by forskolin, but not by neurotrophins or growth factors, in PC12 pheochromocytoma cells. One of these genes, SIK (salt-inducible kinase), is a member of the sucrose-nonfermenting 1 protein kinase/AMP-activated protein kinase protein kinase family that was also recently identified from the adrenal gland of rats treated with high-salt diets. SIK mRNA is induced up to eightfold in specific regions of the hippocampus and cortex in rats, following systemic kainic acid administration and seizure induction.

Prevalence and molecular characterization of Aeromonas spp. in ready-to-eat foods in Italy.

A survey was carried out in Italy to ascertain the prevalence of Aeromonas spp. in ready-to-eat foods (vegetables, cheeses, meat products, and ice creams) and the level of molecular heterogeneity of the isolates found by macrorestriction analysis of genomic DNA with pulsed-field gel electrophoresis (PFGE). In total, 46 (14.4%) of the 320 food samples examined were found positive for Aeromonas spp. The highest percentages of isolation were discovered in vegetables, particularly lettuce (45.0%), endive (40.0%), and rucola (20.0%). Ricotta was the only cheese type analyzed that showed a high frequency of isolation (45.0%). Among meat products, salami and raw ham (25.0% of samples positive) and, to a lesser extent, baloney (5.0%) were found positive for Aeromonas spp. Aeromonas hydrophila was the most common isolate from foods of animal origin, whereas Aeromonas caviae was the dominant species in vegetables. No motile aeromonads were found in ice cream samples. Aeromonas isolates showed a high level of genetic heterogeneity, because 24 PFGE patterns were identified among 27 A. hydrophila strains and 20 PFGE patterns were found in 23 A. caviae isolates. In conclusion, consumers of ready-to-eat foods in Italy are regularly exposed to many genetically distinct strains of A. hydrophila and A. caviae without evident signs of malaise, and therefore, few of these strains, if any, are likely to be pathogenic.

Variations of synaptotagmin I, synaptotagmin IV, and synaptophysin mRNA levels in rat hippocampus during the estrous cycle.

Periodic changes in ovarian steroid levels during fertility cycles affect learning both in humans and in rats in parallel with electrophysiological and morphological fluctuations in selective neuronal populations. In particular, during the estrous cycle of the female rat, hippocampal CA1 region undergoes cyclic modifications in synaptic density. To investigate the molecular mechanisms involved in synaptic remodeling during the estrous cycle, we analyzed the expression of three presynaptic markers, synaptotagmin I, synaptotagmin IV, and synaptophysin, in the female adult rat brain by in situ hybridization. Relative abundance in mRNA for these three markers was quantified at four phases of the estrous cycle: diestrus, proestrus (AM and PM), and estrus. mRNA levels for syt1 exhibited cyclic variations in pyramidal neurons of the CA3 region of hippocampus during the estrous cycle, while mRNA levels for syt4 and SYN were relatively invariant in this or other regions of the hippocampus. Because CA3 pyramidal neurons make synaptic contacts in CA1, modulation of syt1 expression in CA3 may participate in the changes in synaptic density observed in CA1 during the estrous cycle. Furthermore, both syt1 and SYN mRNA varied cyclically in layer II, but not in layer III of entorhinal cortex, while syt4 remained unchanged throughout the cycle. These data suggest that regular variations in steroid hormone levels during fertility cycles may alter the properties of several networks involved in information processing and learning and memory through altered levels of presynaptic proteins.

Protein synthesis in presynaptic endings from squid brain: modulation by calcium ions.

Previous biochemical, autoradiographic, and ultrastructural data have shown that, in the synaptosomal fraction of the squid optic lobe, protein synthesis is largely due to the presynaptic terminals of the retinal photoreceptor neurons (Crispino et al. [1993a] Mol. Cell. Neurosci. 4:366-374; Crispino et al. [1993b] J. Neurochem. 61:1144-1146; Crispino et al. [1997] J. Neurosci. 17:7694-7702). We now report that this process is close to its maximum at the basal concentration of cytosolic Ca++, and is markedly inhibited when the concentration of this ion is either decreased or increased. This conclusion is supported by the results of experiments with: 1) compounds known to increase the level of cytosolic Ca++, such as A23187, ionomycin, thapsigargin, and caffeine; 2) compounds sequestering cytosolic calcium ions such as BAPTA-AM; and 3) agents that block the role of Ca++ as second messenger, such as TFP and W7, which inhibit calmodulin, and calphostin, which inhibits protein kinase C. We conclude that variations in the level of cytosolic Ca++ induced in presynaptic terminals by neuronal activity may contribute to the modulation of the local synthesis of protein.