The autopsy at the time of SARS-CoV-2: Protocol and lessons.

A new viral disease named COVID-19 has recently turned into a pandemic. Compared to a common viral pneumonia it may evolve in an atypical way, causing the rapid death of the patient. For over two centuries, autopsy has been recognized as a fundamental diagnostic technique, particularly for new or little-known diseases. To date, it is often considered obsolete giving the inadequacy to provide samples of a quality appropriate to the sophisticated diagnostic techniques available today. This is probably one of the reasons why during this pandemic autopsies were often requested only in few cases, late and discouraged, if not prohibited, by more than one nation. This is in contrast with our firm conviction: to understand the unknown we must look at it directly and with our own eyes. This has led us to implement an autopsy procedure that allows the beginning of the autopsy shortly after death (within 1-2 h) and its rapid execution, also including sampling for ultrastructural and molecular investigations. In our experience, the tissue sample collected for diagnosis and research were of quality similar to biopsy or surgical resections. This procedure was performed ensuring staff and environmental safety. We want to propose our experience, our main qualitative results and a few general considerations, hoping that they can be an incentive to use autopsy with a new procedure adjusted to match the diagnostic challenges of the third millennium.

Survival following treatment for intracranial ependymoma: a review.

The actual definition of survival rates following treatment for intracranial ependymomas is substantially influenced by the strict interaction among different factors. Age, location, and grading, for example, act together, negatively influencing the prognosis of younger children also invariably influenced by the more demanding role of surgery and the still limited use, up to recently, of radiotherapy under 3 years of age. In the same direction, the worse prognosis in most series of infratentorial ependymomas if compared with their supratentorial counterpart should be cautiously considered, midline posterior fossa tumors having completely different implications from those originating or predominantly extending to the cerebellopontine angle, where the extent of surgery has more invariably to compare with patients' quality of life. New radiotherapic regimens and their applications in infancy are promisingly demonstrating an improvement of present prognostic criteria, with the limit of still insufficient information on their long-term secondary effects. Similarly, molecular biology research studies, though still in their preclinical stage, are prompting to change the concept of a substantially chemoresistant tumor helping to stratify these lesions with the final aim of targeted pharmacological therapies. In the present review paper, we investigated singularly the role that the more commonly considered prognostic factors have had in the literature on survival of children affected by intracranial ependymomas, trying to elucidate their cumulative effect on the actual knowledge of this issue.

End-to-side nerve neurorrhaphy: critical appraisal of experimental and clinical data.

End-to-side neurorrhaphy (ESN) or terminolateral neurorraphy consists of connecting the distal stump of a transected nerve, named the recipient nerve, to the side of an intact adjacent nerve, named the donor nerve, "in which only an epineurial window is performed". This procedure was reintroduced in 1994 by Viterbo, who presented a report on an experimental study in rats. Several experimental and clinical studies followed this report with various and sometimes conflicting results. In this paper we present a review of the pertinent literature. Our personal experience using a sort of end-to-side nerve anastomosis, in which the donor nerve is partially transected, is also presented and compared with ESN as defined above. When the proximal nerve stump of a transected nerve is not available, ESN, which is claimed to permit anatomic and functional preservation of the donor nerve, seems an attractive technique, though yet not proven to be effective. Deliberate axotomy of the donor nerve yields results that are proportional to the entity of axotomy, but such technique, though resembling ESN, is an end-to-end neurorrhaphy. Neither experimental or clinical evidence support liberalizing the clinical use of ESN, a procedure with only an epineurial window in the donor nerve and without deliberate axotomy. Much more experimental investigation needs to be done to explain the ability of normal, intact nerves to sprout laterally. Such procedure appears justified only in an investigational setting.

Linkage analysis of candidate regions using a composite neurocognitive phenotype correlated with schizophrenia.

As schizophrenia is genetically and clinically heterogeneous, systematic investigations are required to determine whether ICD-10 or DSM-IV categorical diagnoses identify a phenotype suitable and sufficient for genetic research, or whether correlated phenotypes incorporating neurocognitive performance and personality traits provide a phenotypic characterisation that accounts better for the underlying variation. We utilised a grade of membership (GoM) model (a mathematical typology developed for studies of complex biological systems) to integrate multiple cognitive and personality measurements into a limited number of composite graded traits (latent pure types) in a sample of 61 nuclear families comprising 80 subjects with ICD-10/DSM-IV schizophrenia or schizophrenia spectrum disorders and 138 nonpsychotic first-degree relatives. GoM probability scores, computed for all subjects, allowed individuals to be partly assigned to more than one pure type. Two distinct and contrasting neurocognitive phenotypes, one familial, associated with paranoid schizophrenia, and one sporadic, associated with nonparanoid schizophrenia, accounted for 74% of the affected subjects. Combining clinical diagnosis with GoM scores to stratify the entire sample into liability classes, and using variance component analysis (SOLAR), in addition to parametric and nonparametric multipoint linkage analysis, we explored candidate regions on chromosomes 6, 10 and 22. The results indicated suggestive linkage for the familial neurocognitive phenotype (multipoint MLS 2.6 under a low-penetrance model and MLS>3.0 under a high-penetrance model) to a 14 cM area on chromosome 6, including the entire HLA region. Results for chromosomes 10 and 22 were negative. The findings suggest that the familial neurocognitive phenotype may be a pleiotropic expression of genes underlying the susceptibility to paranoid schizophrenia. We conclude that use of composite neurocognitive and personality trait measurements as correlated phenotypes supplementing clinical diagnosis can help stratify the liability to schizophrenia across all members of families prior to linkage, allow the search for susceptibility genes to focus selectively on subsets of families at high genetic risk, and augment considerably the power of genetic analysis.

Analysis of the 5' and 3'UTRs in the post-transcriptional regulation of the interleukin-5 gene.

Post-transcriptional regulation is emerging as an important control point in cytokine gene expression. However, the role that it plays in IL-5 gene expression is unclear with some conflicting reports. Here we investigate the importance of post-transcriptional regulation and the role of the 5' and 3' untranslated regions (UTRs) in mIL-5 gene expression. To do this, IL-5 expression from a panel of cDNA constructs was compared. We found it essential to remove the 5' synthetic oligonucleotide tails, introduced during the cloning of the mIL-5 cDNA, when studying IL-5 expression. The presence of these oligo(G) tails acted as potent inhibitors of translation of both SV40 and SP6 transcripts. Furthermore, the length of the tails was found to be critical to the translational efficiency. Taking this into account, we found no evidence to suggest that IL-5 is regulated at the level of mRNA stability or translation efficiency by either the 5' or 3'UTR. These results suggest that post-transcriptional control is not a major factor regulating IL-5 expression.

Identification of two novel palindromic regulatory elements in the murine interleukin-5 promoter.

Interleukin-5 has remarkable specificity for the eosinophil lineage. This fact, combined with the biological specificity of eosinophilia suggests tight and independent regulation of IL-5 expression. Here we report two novel palindromic regulatory elements (PRE) which contain positive regulatory motifs (PRM) that control transcription of the murine IL-5 gene. The first element, mPRE1-IL5 (-79 to -90) contains the mPRM1 at positions -87 to -89 which operates as a positive regulatory element with mutation of this motif resulting in a 64% decrease in gene activity. Gene expression was reduced by 67% when a similar mutation was introduced into the mPRM2 (-467 to -469) of mPRE2-IL5 (-459 to -470). Both elements specifically bind proteins from EL4-23 cell nuclear extracts forming constitutive DNA-protein complexes. EMSA experiments utilising mutated mPRE-IL5 oligonucleotides indicate that in both elements, the mPRMs are essential for protein binding.

Two pathways can activate the interleukin-5 gene and induce binding to the conserved lymphokine element 0.

Eosinophilia is a uniquely specific phenomenon regulated by interleukin-5 (IL-5), suggesting specific control for IL-5 gene expression. Using a transient-transfection reporter assay and DNA mobility-shift experiments in EL4 mouse lymphoma cells, reporter expression and binding of transcription factors to the conserved lymphokine element 0 (CLE0) in the mouse (mIL-5) promoter was investigated. Activation of the IL-5 promoter required costimulation of T cells with phorbol ester (phorbol 12-myristate 13-acetate [PMA]) and cyclic adenosine 3',5'-monophosphate (cAMP), but was blocked by the immunosuppressive drug, cyclosporin A (CsA). Binding to CLE0 was induced under conditions optimal for IL-5 transcription but was not blocked by CsA. CD28-induced signals could partly substitute for cAMP. However, the effects of cAMP, but not of CD28, were sensitive to the cAMP-dependent protein kinase inhibitor, H89, suggesting that CD28 does not involve a cAMP mechanism. It therefore appears that IL-5 expression can be induced by at least two distinct stimulatory pathways. Although CLE0 contains sequences similar to AP-1 and NF-AT, only the AP-1 moiety of the CLE0 element could be demonstrated to have inducible binding. Experiments with antisera to the AP-1 family of transcription factors indicated that c-fos and JunB bind to the IL-5 CLE0 in activated lymphoma cells. The role of the NF-AT-like element was less clear. A constitutively expressed protein showed a weak band that was inhibited by mIL-2 NF-AT competitor sequences. However, this protein did not react with an anti-NF-ATp antiserum. On the other hand, transcription was partially inhibited by an oligonucleotide containing the intact NF-AT-like element from CLE0, suggesting that the element is important for optimal transcription, but the nature of the protein binding to it remains unknown. The fact that these factors are induced in a subclone of EL4 that does not express IL-5 and bind to a number of other cytokine gene promoters suggests that although binding to CLE0 appears to be necessary for IL-5 transcription, other factors must control the specific expression of the gene.

[Strain-gauge plethysmographic measurement of the systemic bioavailability of oral nitroglycerin in liver cirrhosis]. / Misura mediante pletismografia strain-gauge della biodisponibilita' sistemica della trinitrina per os in corso di cirrosi epatica.

7 cirrhotic (M = 3, F = 4, mean age 55, range 35-74) and 7 healthy subjects (M = 6, F = 1, mean age 24, range 23-40) were studied. 2.5mg% nitroglycerin were administered per os. This drug is quite completely metabolized in its first pass through the liver (first pass effect). Peripheric vascular effect of nitroglycerin was evaluated by venous occlusion strain-gauge plethysmography, ECG-coupled (Rest Flow measurement RF, in ml/min/100 ml). No statistically significant differences were found between pre-drug RF in the two groups and between pre and post-drug measurements in healthy subjects. Post-drug RF decreased in cirrhotic subjects when compared either to pre-drug values or to post-drug values in normal subjects (statistically significant after the third minute, p ranging less than 0.05 and less than 0.001). The different peripheric vascular effect found in the two groups was considered as a consequence of the increased drug bioavailability in cirrhotics, caused by portosystemic shunts.