Biomedical subjects
Publications and source records attributed to M Paterlini.
Quick postoperative catheter removal remains one of the main goals of radical prostatectomy, but it leads to a greater risk of urinary leakage. Transrectal ultrasonography with enhancing contrast medium (Levovist) is a simple, effective, and minimally invasive examination to evaluate vesicourethral integrity.
The cerebellum maintains balance and orientation, refines motor action, stores motor memories, and contributes to the timing aspects of cognition. We generated two mouse lines for making Cre recombinase-mediated gene disruptions largely confined to adult cerebellar granule cells. For this purpose we chose the GABA(A) receptor alpha6 subunit gene, whose expression marks this cell type. Here we describe mouse lines expressing Cre recombinase generated by 1) Cre knocked into the native alpha6 subunit gene by homologous recombination in embryonic stem cells; and 2) Cre recombined into an alpha6 subunit gene carried on a bacterial artificial chromosome (BAC) genomic clone. The fidelity of Cre expression was tested by crossing the mouse lines with the ROSA26 reporter mice. The particular alpha6BAC clone we identified will be valuable for delivering other gene products to cerebellar granule cells.
A novel splice variant of metabotropic glutamate receptor type 6 (mGlu6 receptor) was identified by reverse transcription-polymerase chain reaction amplification and sequence analysis of rat retina cDNA. The new rat receptor isoform (mGlu6b receptor) is characterized by an additional exon of 88 nucleotides containing an inframe stop codon, thus predicting the expression of a truncated protein of 508 amino acids. In situ hybridization reveals mGlu6b receptor mRNA to be predominantly expressed in the outer part of the inner nuclear layer of rat retina, containing ON-bipolar cells. The mGlu6b protein would comprise the extracellular domain of the receptor containing the ligand-binding site, but would lack the transmembrane and intracellular portions, thus possibly acting as a retinal soluble receptor for glutamate.
Novel splice variants of metabotropic glutamate receptor type 6 (mGlu6 receptor) were identified by reverse transcription-polymerase chain reaction (RT-PCR) amplification and sequence analysis of rat and human retina cDNAs. The new rat mGlu6 receptor mRNA isoform is characterized by an additional exon of 88 nucleotides containing an in frame stop codon, thus predicting the expression of a truncated protein of 508 amino acids. The human retina was found to express two different mGlu6 receptor mRNA variants: one lacking 97 nucleotides from exon 6, the other including five nucleotides of intron 5. These mRNAs would encode truncated receptors of 425 and 405 amino acids, respectively. Both in rats and in humans, the truncated mGlu6 receptor proteins would comprise the extracellular domain but lack the transmembrane and intracellular portion of the receptor, thus possibly acting as retinal soluble receptors for glutamate. Though generated by different patterns of alternative splicing, the inter-species conservation of truncated mGlu receptor molecules strongly suggest their relevance in the regulatory network of glutamatergic neurotransmission.
PURPOSE: Pelvic abscess after pelvic surgery may be a serious complication. We describe a technique of accurate transrectal ultrasound guided transperineal percutaneous drainage. MATERIAL AND METHODS: Drainage of a pelvic abscess was required in a patient 30 days after radical cystectomy and ileal neobladder construction were performed for bladder cancer. Drainage was performed via perineal access under transrectal ultrasound guidance. RESULTS: The procedure was painless and easily performed. About 100 ml. of pus were removed. The next day the patient was afebrile and 5 days later he was discharged from the hospital. CONCLUSIONS: Pelvic abscess after pelvic surgery is a serious complication. Percutaneous perineal ultrasound guided drainage is a safe and effective therapeutic option.
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The neuronal calcium sensor proteins are members of the calcium-binding protein superfamily. They control localized calcium signalling on membranes and may make G-protein cascades sensitive to cytosolic calcium. The family members are recoverin (visinin, S-modulin), neuronal calcium sensor-1 (frequenin), hippocalcin, neuronal visinin-like protein-1 (visinin-like protein, neurocalcin-alpha), neuronal visinin-like protein-2 and neuronal visinin-like protein-3. Recoverin is expressed only in the retina and pineal gland. Using in situ hybridization, we mapped the expression of the other neuronal calcium sensor protein genes in the adult rat brain. Neuronal visinin-like protein-1 messenger RNA has a widespread distribution and is abundant in all brain areas except the caudate-putamen. Neuronal calcium sensor-1 gene expression is pan-neuronal. Neuronal calcium sensor-1 messenger RNA is present in the dendrites of hippocampal pyramidal and granule cells, suggesting a specific role in dendritic function. Hippocalcin and neuronal visinin-like protein-2 are mainly expressed in the forebrain and have similar expression patterns (neocortex, hippocampus and caudate-putamen). Neuronal visinin-like protein-3 has the most restricted expression; its highest expression level is in the cerebellum (Purkinje and granule cells). However, the neuronal visinin-like protein-3 gene is also expressed in many ventral nuclei throughout the fore- and midbrain, in the medial habenulae, and in the superior and inferior colliculi. The neuronal calcium sensor proteins are a relatively unexplored family of Ca(2+)-binding proteins. They are likely to be involved in many diverse areas of neuronal signalling. In this paper, we describe their expression in the rat brain as determined by in situ hybridization. As all five neuronal calcium sensor protein genes have distinctive expression patterns, they probably perform specific functions.
Human NT2-N neurons derived from retinoic acid treatment of the NTera 2 cell line were used to determine the consequences of ionotropic glutamate receptor (iGluR) hyperstimulation and possible modulatory role(s) exerted by metabotropic glutamate receptor (mGluR) activation. We found that NT2-N neurons express the NR1 subunit of N-methyl-D-aspartate (NMDA) iGluRs and mRNA encoding the 1a isoform of mGluRs. A 15 min pulse with 100 microM NMDA induced an increase in the levels of tau proteins in NT2-N cells. This effect was prevented by incubating NT2-N neurons in the presence of the mGluR agonist (1 S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD). This phenomenon was related, in terms of doses and time, with the observed 1S,3R-ACPD-mediated protection against NMDA-induced NT2-N cell death. Our findings suggest that iGluRs and mGluRs might participate in the control of human neuron viability by differentially affecting the expression of tau proteins.
Sixty-one patients with vesical stones (38 with underlying obstructive conditions and 23 unobstructed) underwent SWL using ultrasound targeting under no regional or general anesthesia. A foley catheter was not routinely employed, and the bladder was filled in a physiologic way. Complete resolution was obtained in 47 patients (78%); in particular, 66% of the obstructed patients and 96% of the unobstructed patients became stone free in one to four SWL sessions. The average number of sessions for all patients was 1.28+/-0.63. Fragments were completely evacuated also in some patients with severe obstruction and in all three patients with neurogenic bladder dysfunction. The size and number of stones did not seem to play a limiting role in SWL effectiveness: the principal limiting factor was the hardness of the stone. No severe complications occurred. However, in six patients (10%), some fragments stopped in the urethra, causing acute urine retention, and endoscopic extraction was necessary. Echo-guided SWL of bladder stones is safe and highly effective in nonobstructed patients and can be considered the elective monotherapy method. In obstructed patients, SWL efficacy is lower, but the method may be suggested for patients who refuse or delay other, more invasive techniques.
It has recently become clear that, analogous to the situation observed in the brain, glutamatergic neurotransmission in the spinal cord relies upon the activation of ionotropic and metabotropic glutamate receptors (mGluRs). Although electrophysiological studies have emphasized the relevance of mGluR activation in the modulation of spinal neuron responses to glutamate, a detailed study of the molecular forms of receptors belonging to the mGluR family in the spinal cord is still lacking. Using a reverse transcriptase-polymerase chain reaction technique with primers specifically recognizing each cloned mGluR subtype, we found that rat adult spinal cord specifically expresses high levels of mRNA encoding mGluR1a but not mGluR1b. The expression of mRNAs for mGluRs 5a and 5b was also found at levels roughly comparable to that observed in mGluR5-positive brain areas. mGluR2, mGluR3, mGluR4 and mGluR7 mRNAs appeared to be expressed at lower levels, while mGluR6 and mGluR8 mRNAs were not detectable.
By combining biochemical, molecular and immunohistochemical approaches, we have investigated the presence of metabotropic glutamate receptors (mGluRs) belonging to the subtype 5 in rat and human spinal cords and the developmental changes in their expression. A polyclonal antibody raised against the carboxy-terminal portion of mGluR5 was used to study the distribution of the receptor in rat foetal (Et15), neonatal (P8) and adult spinal cords and dorsal root ganglia (DRG). mGluR5 appeared to be predominantly expressed in regions containing the primary sensory afferents. Immunoblotting with anti-mGluR5 antibody revealed lower receptor protein levels in rat adult spinal cord when compared with P8 rat spinal cord. Reverse transcriptase-polymerase chain reaction showed both mGluR5a and mGluR5b mRNAs expression in rat spinal cord. The mGluR5a variant was found more abundant in young animals than in adults. The pattern of mGluR5 immunostaining was also studied in foetal (6-8, 10, 12 and 22 weeks of gestation) and adult human spinal cord. At all stages of human development, a strong mGluR5 immunoreactivity was observed in the dorsal roots and in the dorsal and dorsolateral funiculi with maximum levels of staining at week 12 of gestation. Foetal DRG neurons were heterogeneously labeled. mGluR5 was also diffusely detectable in the mantle layer. In adult human spinal cords, immunoreactivity was confined to laminae I and II of the dorsal horns. These results demonstrate for the first time the presence of mGluR5 in human spinal cord. The distribution of this receptor suggests a role in the development of somatosensory pathways and in the control of nociceptive neurotransmission.
The effects of the ionotropic glutamate receptor (iGluR) selective agonist N-methyl-D-aspartate (NMDA) on amyloid precursor protein (APP) levels were investigated in primary cultures of rat cerebellar granule cells. Both immunocytochemistry and immunoblotting techniques showed increased APP levels 4 h after a 15 min pulse with NMDA. This effect was completely prevented by incubating the neurones in the presence of the selective metabotropic GluR (mGluR) agonist 1S,3R-ACPD. This phenomenon was related, in terms of doses and time, with the observed 1S,3R-ACPD-mediated protection on NMDA-induced granule cell death. Our findings indicate that APP metabolism is differentially regulated by the stimulation of various GluR subtypes. The GluR-mediated changes in APP content might participate in the control of neuronal viability.