Prognostic MRI parameters in acute traumatic cervical spinal cord injury.

PURPOSE: The aim of this study is to estimate the prognostic value of some features documented on preoperative MRI study in patients with acute cervical spinal cord injury. METHODS: The study was conducted in patients operated for cervical spinal cord injury (cSCI) from April 2014 to October 2020. The quantitative analysis on preoperative MRI scans included: length of the spinal cord intramedullary lesion (IMLL the canal diameter at the level of maximal spinal cord compression (MSCC) and the presence of intramedullary hemorrhage. The canal diameter at the MSCC was measured on the middle sagittal FSE-T2W images at the maximum level of injury. The America Spinal Injury Association (ASIA) motor score was used for neurological assessment at hospital admission. At 12-month follow-up all patients were examined with the SCIM questionnaire. RESULTS: At linear regression analysis, the length of the spinal cord lesion [ß coefficient -10.35, 95% confidence interval (CI)-13.71 to-6.99; p < 0.001], the diameter of the canal at the level of the MSCC (ß coefficient 6.99, 95% CI 0.65 to 13.33; p = 0.032), and the intramedullary hemorrhage (ß coefficient - 20.76, 95% CI - 38.70 to - 2.82; p = 0.025), were significantly associated with the score at the SCIM questionnaire at one year follow-up: shorter spinal cord lesion, greater diameter of the canal at the level of the MSCC, and absence of intramedullary hemorrhage were predictors of better outcome. CONCLUSION: According to the findings of our study, the spinal length lesion, canal diameter at the level of spinal cord compression and intramedullary hematoma documented by the preoperative MRI study were associated with the prognosis of patients with cSCI.

DNAscan: personal computer compatible NGS analysis, annotation and visualisation.

BACKGROUND: Next Generation Sequencing (NGS) is a commonly used technology for studying the genetic basis of biological processes and it underpins the aspirations of precision medicine. However, there are significant challenges when dealing with NGS data. Firstly, a huge number of bioinformatics tools for a wide range of uses exist, therefore it is challenging to design an analysis pipeline. Secondly, NGS analysis is computationally intensive, requiring expensive infrastructure, and many medical and research centres do not have adequate high performance computing facilities and cloud computing is not always an option due to privacy and ownership issues. Finally, the interpretation of the results is not trivial and most available pipelines lack the utilities to favour this crucial step. RESULTS: We have therefore developed a fast and efficient bioinformatics pipeline that allows for the analysis of DNA sequencing data, while requiring little computational effort and memory usage. DNAscan can analyse a whole exome sequencing sample in 1 h and a 40x whole genome sequencing sample in 13 h, on a midrange computer. The pipeline can look for single nucleotide variants, small indels, structural variants, repeat expansions and viral genetic material (or any other organism). Its results are annotated using a customisable variety of databases and are available for an on-the-fly visualisation with a local deployment of the gene.iobio platform. DNAscan is implemented in Python. Its code and documentation are available on GitHub: https://github.com/KHP-Informatics/DNAscan . Instructions for an easy and fast deployment with Docker and Singularity are also provided on GitHub. CONCLUSIONS: DNAscan is an extremely fast and computationally efficient pipeline for analysis, visualization and interpretation of NGS data. It is designed to provide a powerful and easy-to-use tool for applications in biomedical research and diagnostic medicine, at minimal computational cost. Its comprehensive approach will maximise the potential audience of users, bringing such analyses within the reach of non-specialist laboratories, and those from centres with limited funding available.

The diimide drug PIPER has a cytotoxic dose-dependent effect in vitro and inhibits telomere elongation in HELA cells.

It is known that, in vitro, PIPER (N,N'-bis [2-(1-piperidino)ethyl]-3,4,9,10-tetracarboxylic diimide) induces the formation of the Hoogsteen quadruplex structure in telomere DNA, thus inhibiting the polymerisation of telomeric repeats. Since the action of PIPER in vivo has been scarcely investigated, this study was addressed to gain some insight into the effects of this drug on cultured HeLa cells. Vital staining with erythrosine, performed on cells exposed to different PIPER concentrations (from 1 to 50 microM), showed that the drug exerts a dose-dependent cytotoxic effect, clearly evident after a short-term (24 h) treatment. This early cytotoxic effect of PIPER on cultured HeLa cells was confirmed by a spectrophotometric/colorimetric method employing methylthiazoletetrazolium (Mossmann assay). Hematoxylin/eosin staining of cells treated with PIPER for 24 h showed a nuclear condensation and a cytoplasmic vacuolisation, very pronounced at higher drug concentrations. These pictures suggest that PIPER-induced cell death might be of the apoptotic type. Finally, the anti-telomerase activity of PIPER was monitored by TRAP assay, performed on HeLa cell nuclear extracts treated with increasing drug concentrations. It was found that some inhibition of telomerase is apparent even at low concentrations, while at the highest concentration the enzyme is completely inhibited. These results indicate that the cytotoxic power of PIPER is possibly related to its antitelomeric effect.

Variations of telomerase activity in cultured mouse fibroblasts upon proliferation of polyomavirus.

Telomerase plays a central role in various biological phenomena such as cell differentiation and proliferation, apoptosis, malignant transformation and virus infection, for instance HIV and papillomavirus. In addition, it has recently been shown that, in human fibroblasts transformed by monkey polyomavirus SV40, telomeres became stabilized as a consequence of telomerase activation. However, no information exists on the effects of acute infection by murine polyomavirus on the telomeres maintenance and telomerase activity in the host cell. In this paper we report on a differential activity of telomerase in productively infected cells. The results showed a decreased activity of the enzyme as assessed by the TRAP assay. The decrease had already occurred at a non-lytic time of infection and was observed both after infection and naked DNA transfection. Therefore nuclear decapsidation is not involved in the determination of the phenomenon that is attributed to the proliferation of the virus.

Molecular characterization and action of usnic acid: a drug that inhibits proliferation of mouse polyomavirus in vitro and whose main target is RNA transcription.

Usnic acid is a normal component of lichen cells. This natural compound has shown different biological and physiological activities that might have a great relevance in pharmacology and clinics. For instance, usnic acid is known for its antibacterial and antiparasitic action. Also, the drug has a potential interest in cancer therapy because of its antimitotic and antiproliferative action. The molecular structure of usnic acid has been validated and further explored in this investigation. Many biological properties of this drug are known; however its potential antiviral action has not yet been evaluated. In this paper, we demonstrate that usnic acid is a potent inhibitor of the proliferation of mouse polyomavirus. Its action is not exerted at the level of virion entry into the host cell. Moreover, the abolition of viral DNA replication is an indirect consequence of the drastic inhibition of RNA transcription.

Ubiquitin dependent proteolysis is activated in apoptotic fibroblasts in culture.

The ubiquitin mediated pathway constitutes an early response in cultured cells where apoptosis, assessed by internucleosomal specific DNA fragmentation, was induced by serum withdrawal. Data demonstrate that nuclear ubiquitin proteolytic system, but not cytoplasmic, is activated. This activation is paralleled by a substantial chromatin de-condensation. We suggest that chromatin relaxation is causative of the fragmentation since it exposes the DNA to nucleolytic attack. Finally, maintenance of homeostasis and induction of apoptosis seem to undergo a parallel contemporary pathway with a possible mutual feedback.

Cosmology from MAXIMA-1, BOOMERANG, and COBE DMR cosmic microwave background observations.

Recent results from BOOMERANG-98 and MAXIMA-1, taken together with COBE DMR, provide consistent and high signal-to-noise measurements of the cosmic microwave background power spectrum at spherical harmonic multipole bands over 2

Measurement of a Peak in the Cosmic Microwave Background Power Spectrum from the North American Test Flight of Boomerang.

We describe a measurement of the angular power spectrum of anisotropies in the cosmic microwave background (CMB) at scales of 0&fdg;3 to 5 degrees from the North American test flight of the Boomerang experiment. Boomerang is a balloon-borne telescope with a bolometric receiver designed to map CMB anisotropies on a long-duration balloon flight. During a 6 hr test flight of a prototype system in 1997, we mapped more than 200 deg(2) at high Galactic latitudes in two bands centered at 90 and 150 GHz with a resolution of 26&arcmin; and 16&farcm;5 FWHM, respectively. Analysis of the maps gives a power spectrum with a peak at angular scales of 1 degrees with an amplitude 70 µK(CMB).

A flat Universe from high-resolution maps of the cosmic microwave background radiation

The blackbody radiation left over from the Big Bang has been transformed by the expansion of the Universe into the nearly isotropic 2.73 K cosmic microwave background. Tiny inhomogeneities in the early Universe left their imprint on the microwave background in the form of small anisotropies in its temperature. These anisotropies contain information about basic cosmological parameters, particularly the total energy density and curvature of the Universe. Here we report the first images of resolved structure in the microwave background anisotropies over a significant part of the sky. Maps at four frequencies clearly distinguish the microwave background from foreground emission. We compute the angular power spectrum of the microwave background, and find a peak at Legendre multipole Ipeak = (197 +/- 6), with an amplitude delta T200 = (69 +/- 8) microK. This is consistent with that expected for cold dark matter models in a flat (euclidean) Universe, as favoured by standard inflationary models.

The ionophore monensin inhibits mouse polyomavirus DNA replication and destabilizes viral early mRNAs.

Monensin is a ionophore compound with different biological activities. It raises the intralysosomal pH, it binds the plasma membranes particularly at the level of the cisternal system of the Golgi apparatus. It causes imbalance in the intramembrane ion traffic and inhibits export of secretory proteins at membrane level. Monensin blocks endocytosis and therefore impedes entry of toxic molecules. The drug also inhibits viral proliferation of RNA and DNA viruses such as vesicular stomatitis, influenza and human polyomaviruses. In this report we show that monensin effectively abolishes viral DNA replication of mouse polyomavirus. Results show that the half life of viral early mRNAs is significantly reduced in the presence of the drug. Therefore we suggest that the reduction of viral DNA synthesis is a consequence of the reduced intranuclear pool of viral early antigens.

Apoptosis dependent decrease of the intramembrane ion traffic in cultured mouse fibroblasts shown by conductivity dispersion.

We have investigated the intramembranal ion traffic in apoptotic 3T6 cells in culture. Apoptosis was induced by various treatments, such as serum deprivation, high density growth and hydrogen peroxide at subnecrotic doses. Cell death was assessed by nucleosomal DNA fragmentation, single cell electrophoresis, immunofluorescence and histological staining. To study the modifications of membrane structure and function, we adopted a well established biophysical strategy based on the measurement of the electrical conductivity of cell suspensions, as a function of the frequency of the electrical field applied to the sample. A comparison between the conductivity of normal and apoptotic cell suspensions shows that programmed cell death causes a decrease of membrane conductivity which indicates a diminished intramembranal ion traffic. Our results strongly suggest that one of the early events in the triggering of apoptosis is represented by an overall reduction of plasma membrane function. Finally, our results are in agreement with the idea that the nucleus is not the sole target of the apoptotic process.

Electrical conductivity dispersion as a probe of membrane modifications in mouse polyomavirus infected cells in culture.

In this report we investigate the inhibition of membrane conductivity, due to the murine polyomavirus infection in permissive cells in culture. We define experimental conditions to have reproducible results and demonstrate that the intensity of the effects on the cell membrane, depends upon the virus titer used in the infection. Finally, the virus dependent effects disappear if the infection is performed in the presence of a drug that inhibits polymavirus DNA replication.

Role of mouse polyomavirus late region in the control of viral DNA replication: a review.

The genome of polyomaviruses is divided into two coding regions: the early and the late region. A relatively short regulatory sequence, encompassing the origin of viral DNA replication (ori), separates the two regions encoding the structural genes. In mouse polyomavirus (Py) in particular, the early DNA codes for three antigens: large, middle and small T-antigen (L-T, M-T and S-T, respectively). Large T antigen binds ori and thus regulates both viral DNA transcription and replication. Middle T antigen has been shown to mediate malignant transformation in non-permissive cells in vitro. No defined function has been assigned to the small T antigen although this gene product is thought to act synergistically both with L- and M-T antigens. The viral late region of Py encodes also three different genes whose products form the viral capsid during the productive infection cycle in permissive cells. Py early region was thought to be the only part of the genome necessary to code for proteins of functional and regulatory significance. The viral late region, on the other hand, was for a long time considered a simple reservoir of structural information, since it codes for capsid proteins and was supposedly devoid of functional control properties. This short review is focused on recent works from our and other laboratories, reporting evidence that in Py also the late region has a functional role since late sequences are involved in the control of viral DNA replication and in capsid assembly. Results indicating that this might be true for the cognate simian virus SV40 will be also reviewed.