Long-term cognitive deficits after traumatic brain injury associated with microglia activation.

Traumatic Brain Injury (TBI) is the most prevalent of all head injuries. Microglia play an essential role in homeostasis and diseases of the central nervous system. We hypothesize that microglia may play a beneficial or detrimental role in TBI depending on their state of activation and duration. In this study, we evaluated whether TBI results in a spatiotemporal change in microglia phenotype and whether it affects sensory-motor or learning and memory functions in male C57BL/6 mice. We used a panel of neurological and behavioral tests and a multi-color flow cytometry-based data analysis followed by unsupervised clustering to evaluate isolated microglia from injured brain tissue. We characterized several microglial phenotypes and their association with cognitive deficits. TBI results in a spatiotemporal increase in activated microglia that correlated negatively with spatial learning and memory at 35 days post-injury. These observations could define therapeutic windows and accelerate translational research to improve patient outcomes.

Validity and reliability of different techniques of neck-shaft angle measurement.

AIM: To determine a valid and reliable neck-shaft angle (NSA) measurement method while rotating the pelvises in increments of 5° in order to simulate patient malpositioning. MATERIALS AND METHODS: CT images of 17 patients were used to produce digitally reconstructed radiographs in frontal and lateral views and three-dimensional (3D)-reconstructions of the femurs, considered to be the reference standard. Malpositioning was simulated by axially rotating the frontal radiographs from 0° to 20°. Three operators measured in two-dimensions the NSA using four different methods, three times each, at each axial rotation (AR) position. Method 1 (femoral neck axis drawn by joining the centre of the femoral head (CFH) to the median of the femoral neck base; femoral diaphysis axis drawn by joining the median of two lines passing through the medial and lateral edges of the femoral axis below the lesser trochanter) and method 2 (femoral axis taken as the median of a triangle passing through base of femoral neck and medial and lateral head-neck junction; femoral diaphysis as previous) were described for the first time; method 3 was based on a previous study; method 4 was a free-hand technique. Reliability, validity, and global uncertainty were assessed. RESULTS: Method 1 showed the best reliability and validity. The global uncertainty also showed minimal values for method 1, ranging from 7.4° to 14.3° across AR positions. CONCLUSION: Method 1, based on locating the CFH, was the most reliable and valid method and should be considered as a standardised two-dimensional NSA measurement method for clinical application.

SARS-CoV-2 involvement in central nervous system tissue damage.

As the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread globally, it became evident that the SARS-CoV-2 virus infects multiple organs including the brain. Several clinical studies revealed that patients with COVID-19 infection experience an array of neurological signs ranging in severity from headaches to life-threatening strokes. Although the exact mechanism by which the SARS-CoV-2 virus directly impacts the brain is not fully understood, several theories have been suggested including direct and indirect pathways induced by the virus. One possible theory is the invasion of SARS-CoV-2 to the brain occurs either through the bloodstream or via the nerve endings which is considered to be the direct route. Such findings are based on studies reporting the presence of viral material in the cerebrospinal fluid and brain cells. Nevertheless, the indirect mechanisms, including blood-clotting abnormalities and prolonged activation of the immune system, can result in further tissue and organ damages seen during the course of the disease. This overview attempts to give a thorough insight into SARS-CoV-2 coronavirus neurological infection and highlights the possible mechanisms leading to the neurological manifestations observed in infected patients.

Exosomes From Subjects With Multiple Sclerosis Express EBV-Derived Proteins and Activate Monocyte-Derived Macrophages.

OBJECTIVE: To investigate in a cross-sectional study the effect of serum-derived exosomes on primary human blood monocyte-derived macrophages (MDMs) comparing exosomes from healthy donors vs patients with relapsing-remitting multiple sclerosis in remission and in relapse and to assess whether the response correlates with exosomal Epstein-Barr virus (EBV) protein expression. METHODS: A total of 45 serum-derived exosome preparations were isolated from patients and healthy controls and verified for the expression of exosomal and EBV markers. MDMs were differentiated from monocytes for 7 days and incubated for 24 hours with exosomes, and then, cell supernatants were collected for cytokine measurement by cytometric bead array. Cells were immunophenotyped before and after differentiation. RESULTS: Serum-derived exosomes of patients with multiple sclerosis (MS) expressed higher levels of EBV proteins than healthy controls. Of interest, expression of EBV nuclear antigen EBNA1 and latent membrane proteins LMP1 and 2A was higher on exosomes derived from patients with active RRMS compared with healthy controls and stable patients. After data normalization, we observed that incubation with EBV(+) exosomes induced CXCL10 and CCL2 secretion by MDMs. MDMs differentiated from patients with active disease were better secretors of CXCL10 and other interferon-γ-inducible chemokines, including CCL2 and CXCL9, than MDMs from healthy and stable MS groups. MDMs from active patients had a higher frequency of a CD14(++) subset that correlated with the secreted CXCL10. CONCLUSION: Exosomes expressing EBV proteins correlate with disease activity and induce an inflammatory response in MDMs that is compounded by the origin of the responder cells.

Trypanosoma-Cruzi Cross-Reactive Antibodies Longitudinal Follow-Up: A Prospective Observational Study in Hematopoietic Stem Cell Transplantation.

Antibodies named TcCRA "Trypanosoma cruzi Cross Reactive Antibodies" were detected in 47% of blood donors from French population unexposed to the parasite. In order to evaluate the passive or active transmissibility of TcCRA and further characterize its role and etiology, we have conducted a study in a cohort of 47 patients who underwent allogeneic Hematopoietic Stem Cell Transplantations (allo-HSCT). Donors and recipients were tested for TcCRA prior to transplantation. Recipients were further tested during follow-up after transplantation. Demographical, clinical and biological data were collected. Our primary end-point was to assess the risk of TcCRA acquisition after transplantation. During this initial analysis, we observed no seroconversion in patients receiving cells from TcCRA negative donors (n = 23) but detected seroconversion in 4 out of 24 patients who received hematopoietic stem cells from positive donors. Here, we are discussing possible scenarios to explain TcCRA-immune status in recipient after transplantation.

Anti-ICAM-1 antibody modulates late onset of acinar cell apoptosis and early necrosis in taurocholate-induced experimental acute pancreatitis.

The hallmark of severe acute pancreatitis (SAP) is massive acinar cell death by necrosis. However, programmed, apoptotic acinar cell death has also been observed. Little is known about the dynamics, localization, and inductive factors of acinar cell apoptosis in SAP. We therefore induced SAP in rats by retrograde infusion of 3% sodium taurocholate. Starting as early as 5 minutes after taurocholate administration, small scattered groups of acinar cells showed zymogen degranulation, loss of cell polarity, cytoplasmic microvacuolization, and nuclear shrinkage, but no DNA degradation, thus featuring necrosis. The areas of necrotic acini extended at later time points giving rise to larger areas of complete parenchymal breakdown after 6 hours. Parenchymal degradation was paralleled by neutrophil infiltration and significant tumor necrosis factor (TNF)-alpha mRNA up-regulation. Up to the 12-hour interval, apoptotic acinar cells detected by TUNEL were as rare as in healthy pancreata. At 24 hours, however, the acinar apoptotic rate in nonnecrotic parenchyma had dramatically increased. Pretreatment of rats with anti-ICAM-1 antibody prior to pancreatitis induction led to a significant reduction of neutrophil infiltration along with decreased TNF-alpha mRNA expression throughout the 24-hour observation period without affecting the presence and dynamics of necrosis. However, anti-ICAM-1 pretreatment decreased the extent of acinar cell damage by necrosis and extensively suppressed acinar cell apoptosis. We conclude that taurocholate induces two sequential patterns of acinar cell death in terms of very early necrosis followed by late apoptosis during the postacute phase of SAP. The progression of necrosis and the late apoptotic acinar cell death seem to be influenced by the local presence of neutrophils via a TNF-alpha-dependent mechanism. In addition to augmenting necrosis, neutrophils might have an apoptosis-inducing potential in SAP.

Analysis of forces developed during obturations. Comparison of three gutta-percha techniques.

The aim of this study was to compare the forces developed during three obturation techniques: warm vertical compaction (WVC), lateral condensation (LC), and thermomechanical compaction (TMC). A force-analyzer device allowed us to record and analyze the forces exerted during the obturations. The different methods, which varied in instrumentation, duration, and compaction load, were studied from graphs representing the force application sequences. In addition, the graphs resulting from specific modifications in the LC technique (use of a single spreader) and deliberately induced failures with TMC (with a too-small condensor or with excessive load) were analyzed. The mean values for the vertical forces for WVC, LC, and TMC were, respectively, 2.5 +/- 0.4 kg, 1.1 +/- 0.3 kg, and 1.65 +/- 0.2 kg. The mean values for the lateral forces were, respectively, 0.85 +/- 0.2 kg, 0.35 +/- 0.1 kg, and 0.5 +/- 0.2 kg. The use of graphs provides a new approach to the analysis of obturation methods and may find its greatest application in the teaching of these techniques. It also has the potential to improve research into obturation technique.

A comparative study of four coronal obturation materials in endodontic treatment.

The aim of this study was to compare, in vitro, the ability of temporary versus permanent materials to seal the access cavity. Eighty human maxillary single-canal teeth were prepared biomechanically and obturated with gutta-percha and an endodontic cement AH Plus, using the warm vertical compaction technique. All access cavities were sealed with 1 of 4 materials (Cavit, Fermit, Tetric, or Dyract). Microleakage was assessed by methylene blue dye penetration. The teeth were submitted to 100 thermocycles, with temperature varying from 0 degree to 55 degrees C. The greatest degree of leakage was observed with the temporary materials (Cavit and Fermit). There was a significant difference (p < 0.05) in leakage between all materials except between Dyract and Tetric. This suggests that it may be more prudent to use a permanent restorative material for provisional restorations to prevent inadequate canal sealing and the resulting risk of fluid penetration.

Effect of masticatory cycles on tooth compression and resultant leakage of amalgam retrofills.

This study was carried out to investigate the effect of changes in tooth structure due to masticatory cycles on amalgam retrofillings performed to different cavity depths. Forty upper maxillary first molars were tested. In the obturated palatal root apical amalgam was inserted to different depths (1.5 and 3 mm). Eight casts were built, each having four sample molars. To block the teeth, the vestibular roots were embedded in resin (palatal root was left free). The eight remaining retrofilled teeth served as controls and were not submitted to occlusal forces. A mechanical device to simulate masticatory cycles subjected the teeth to 500,000 to three million cycles. Leakage was assessed from dye penetration observations. The values of microleakage were analyzed and compared, and strain gauges were used to assess structural modifications to the tooth. The leakage of all retrofilled obturations increased in correlation with the number of masticatory cycles. At three million cycles, root length compression was 0.3 +/- 0.02%. Leakage was significantly less for the deeper cavity preparations at all stages. This in vitro study suggests a significant effect by root compression due to masticatory loads on the leakage of retrofilled amalgam obturations.

Effect of masticatory cycles on apical leakage of obturated teeth.

This study investigated the effect of apical leakage due to masticatory cycles on root canal treatment. Twenty upper maxillary molars were first obturated using the warm vertical compaction technique. Four maxillary casts were then built, with each holding four of the sample molars. The molars were embedded in resin with the roots separated from the resin by means of a light silicon. The four remaining teeth served as controls and were not submitted to occlusal forces. A mechanical device to simulate masticatory cycles subjected the teeth to 0.5 x 10(6) cycles (group A), 10(6) cycles (group B), 2 x 10(6) cycles (group C), and 3 x 10(6) cycles (group D); the control was group E. The roots were placed in 2% methylene blue dye solution for 72 h and then sectioned longitudinally so that dye penetration could be measured. The mean values of dye penetration were: 3.70 +/- 0.69 mm, group A; 5.00 +/- 1.14 mm, group B; 6.00 +/- 1.01 mm, group C; 7.23 +/- 0.66 mm, group D; and 2.74 +/- 0.75 mm, group E. The value of dye penetration increased in correlation with the number of masticatory cycles. This in vitro study suggests the significant effect of masticatory loads on apical leakage.

Anti-Trypanosoma cruzi cross-reactive antibodies detected at high rate in non-exposed individuals living in non-endemic regions: seroprevalence and association to other viral serologies.

Cross-reactive antibodies are characterized by their recognition of antigens that are different from the trigger immunogen. This happens when the similarity between two different antigenic determinants becomes adequate enough to enable a specific binding with such cross-reactive antibodies. In the present manuscript, we report the presence, at an "abnormal" high frequency, of antibodies in blood samples from French human subjects cross-reacting with a synthetic-peptide antigen derived from a Trypanosoma cruzi (T. cruzi) protein sequence. As the vector of T. cruzi is virtually confined to South America, the parasite is unlikely to be the trigger immunogen of the cross-reactive antibodies detected in France. At present, the cross-reactive antibodies are measured by using an in-house ELISA method that employs the T. cruzi -peptide antigen. However, to underline their cross-reactive characteristics, we called these antibodies "Trypanosoma cruzi Cross Reactive Antibodies" or TcCRA. To validate their cross-reactive nature, these antibodies were affinity-purified from plasma of healthy blood donor and were then shown to specifically react with the T. cruzi parasite by immunofluorescence. Seroprevalence of TcCRA was estimated at 45% in serum samples of French blood donors while the same peptide-antigen reacts with about 96% of T. cruzi -infected Brazilian individuals. In addition, we compared the serology of TcCRA to other serologies such as HSV 1/2, EBV, HHV-6, CMV, VZV, adenovirus, parvovirus B19, mumps virus, rubella virus, respiratory syncytial virus, measles and enterovirus. No association was identified to any of the tested viruses. Furthermore, we tested sera from different age groups for TcCRA and found a progressive acquisition starting from early childhood. Our findings show a large seroprevalence of cross-reactive antibodies to a well-defined T. cruzi antigen and suggest they are induced by a widely spread immunogen, acquired from childhood. The etiology of TcCRA and their clinical relevance still need to be investigated.

Analysis of forces developed during root canal preparation with the balanced force technique.

The aim of this study was to examine the forces and torque developed during root canal preparation with the balanced force technique using a recently described force-analyser device. A tooth was placed in a holder within the Endograph and forces and torques exerted were recorded. These parameters, which can be studied during preparation (on-line) or stored and examinated subsequently (off-line) generated endograms, which showed the forces generated with time. In addition, the endograms of preparations performed by students and endodontists, as well as deliberately induced failures in preparation technique (broken instruments), were compared. The values for the forces and torques depended on the size of the instruments and were related to the phase of the preparation. For the endodontists, the vertical and horizontal forces varied, respectively, from 0.08 +/- 0.01 kg for a size 15 to 0.65 +/- 0.10 kg for a size 45, and from 0.01 +/- 0.005 kg for a size 15 to 0.4 +/- 0.1 kg for a size 40. The torque varied from 0.08 +/- 1 kg mm-1 for a size 15 to 1.6 +/- 0.4 kg mm-1 for a size 45. With the endograms used as a reference, the relation between the developed vertical forces and the torque became more similar between the groups of endodontists and students. The Endograph provides a new approach to the analysis of preparation technique because it depicts the relationships between the different parameters of the preparation.