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International audience; Safe and effective vaccines against coronavirus disease 2019 (COVID-19) are essential for ending the ongoing pandemic. Although impressive progress has been made with several COVID-19 vaccines already approved, it is clear that those developed so far cannot meet the global vaccine demand alone. We describe a COVID-19 vaccine based on a replication-defective gorilla adenovirus expressing the stabilized prefusion severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein named GRAd-COV2. We assessed the safety and immunogenicity of a single-dose regimen of this vaccine in healthy younger and older adults to select the appropriate dose for each age group. For this purpose, a phase 1, dose-escalation, open-labeled trial was conducted including 90 healthy participants (45 aged 18 to 55 years old and 45 aged 65 to 85 years old) who received a single intramuscular administration of GRAd-COV2 at three escalating doses. Local and systemic adverse reactions were mostly mild or moderate and of short duration, and no serious adverse events were reported. Four weeks after vaccination, seroconversion to spike protein and receptor binding domain was achieved in 43 of 44 young volunteers and in 45 of 45 older participants. Consistently, neutralizing antibodies were detected in 42 of 44 younger-age and 45 of 45 older-age volunteers. In addition, GRAd-COV2 induced a robust and T helper 1 cell (T H 1)-skewed T cell response against the spike protein in 89 of 90 participants from both age groups. Overall, the safety and immunogenicity data from the phase 1 trial support the further development of this vaccine.

Safe and effective vaccines against coronavirus disease 2019 (COVID-19) are urgently needed to control the ongoing pandemic. Although impressive progress has been made with several COVID-19 vaccines already approved, it is clear that those developed so far cannot meet the global vaccine demand. We have developed a COVID-19 vaccine based on a replication-defective gorilla adenovirus expressing the stabilized pre-fusion SARS-CoV-2 Spike protein, named GRAd-COV2. We aimed to assess the safety and immunogenicity of a single-dose regimen of this vaccine in healthy younger and older adults to select the appropriate dose for each age group. To this purpose, a phase 1, dose-escalation, open-label trial was conducted including 90 healthy subjects, (45 aged 18-55 years and 45 aged 65-85 years), who received a single intramuscular administration of GRAd-CoV2 at three escalating doses. Local and systemic adverse reactions were mostly mild or moderate and of short duration, and no serious AE was reported. Four weeks after vaccination, seroconversion to Spike/RBD was achieved in 43/44 young volunteers and in 45/45 older subjects. Consistently, neutralizing antibodies were detected in 42/44 younger age and 45/45 older age volunteers. In addition, GRAd-COV2 induced a robust and Th1-skewed T cell response against the S antigen in 89/90 subjects from both age groups. Overall, the safety and immunogenicity data from the phase 1 trial support further development of this vaccine.One Sentence SummaryGRAd-COV2, a candidate vaccine for COVID-19 based on a novel gorilla adenovirus, is safe and immunogenic in younger and older adults

The COVID-19 pandemic caused by the emergent SARS-CoV-2 coronavirus threatens global public health and there is an urgent need to develop safe and effective vaccines. Here we report the generation and the preclinical evaluation of a novel replication-defective gorilla adenovirus-vectored vaccine encoding the pre-fusion stabilized Spike (S) protein of SARS-CoV2. We show that our vaccine candidate, GRAd-COV2, is highly immunogenic both in mice and macaques, eliciting both functional antibodies which neutralize SARS-CoV-2 infection and block Spike protein binding to the ACE2 receptor, and a robust, Th1-dominated cellular response. We show here that the pre-fusion stabilized Spike antigen is superior to the wild type in inducing ACE2-interfering, SARS-CoV2 neutralizing antibodies. To face the unprecedented need for vaccine manufacturing at massive scale, different GRAd genome deletions were compared to select the vector backbone showing the highest productivity in stirred tank bioreactors. This preliminary dataset identified GRAd-COV2 as a potential COVID-19 vaccine candidate, supporting the translation of GRAd-COV2 vaccine in a currently ongoing Phase I clinical trial (NCT04528641)., Graphical Abstract, Safe and effective vaccines are needed to fight COVID-19 pandemic. Here we report the preclinical evaluation of GRAd-COV2, a replication-defective adenovirus vector derived from a gorilla and encoding the pre-fusion stabilized Spike protein of SARS-CoV2. GRAd-COV2 is currently being tested in Phase 2/3 clinical studies.

Microglia are highly motile glial cells that are proposed to mediate synaptic pruning during neuronal circuit formation. Disruption of signaling between microglia and neurons leads to an excess of immature synaptic connections, thought to be the result of impaired phagocytosis of synapses by microglia. However, until now the direct phagocytosis of synapses by microglia has not been reported and fundamental questions remain about the precise synaptic structures and phagocytic mechanisms involved. Here we used light sheet fluorescence microscopy to follow microglia-synapse interactions in developing organotypic hippocampal cultures, complemented by three-dimensional ultrastructural characterization using correlative light and electron microscopy (CLEM). Our findings define a set of dynamic microglia-synapse interactions, including the selective partial phagocytosis, or trogocytosis (trogo-: nibble), of presynaptic structures and the induction of postsynaptic spine head filopodia by microglia. These findings allow us to propose a mechanism for the facilitatory role of microglia in synaptic circuits remodeling and maturation.

The aim of this study was to evaluate the results of rehabilitative treatment of incontinent patients with anal sphincter lesions in order to verify whether these lesions influence the outcome of rehabilitation.Between January 2008 and June 2011, 48 patients [36 women, age range 51-77 years; 12 men, age range 57-70 years] affected by fecal incontinence were included in the study. After a preliminary clinical evaluation, including the Fecal Incontinence Severity Index (FISI) score, all patients were studied by means of endoanal ultrasound and anorectal manometry. Two patients groups were identified: Group 1 comprised 27 patients with sphincter lesions and Group 2 had 21 patients without sphincter lesions. All 48 underwent a multimodal rehabilitation program. At the end of the rehabilitation program, all patients were reassessed by means of clinical evaluation and anorectal manometry and their data were analyzed.Although we obtained an overall significantly lower FISI score in all patients after rehabilitation (p0.001) when compared with pre-treatment values, our study revealed that patients with sphincter lesions had a significantly worse post-rehabilitative FISI score (p0.003) when compared with those patients with intact anal sphincters and that there is a linear relationship between post-rehabilitative FISI scores and severity of sphincter lesions (ρ s 0.69).These data suggest that rehabilitation may be less effective in patients affected by sphincter defects.

Rehabilitation is the first therapeutic step of obstructed defecation, after failure of conservative therapy with high-fiber diet and laxatives. This study evaluates the usefulness of psyllium, a bulk-forming agent, when used during rehabilitation of obstructed defecation.Between January 2008 and December 2010, 45 patients affected by obstructed defecation were included in the study. Two randomized groups were selected. Group 1 (21 women; age range 25-67 (mean, 51.8) years) continued to consume a high-fiber diet (approximately 30 g fiber per day) during rehabilitation. Group 2 (24 women; age range 46-71 (mean, 59.8) years) consumed only psyllium (3.6 g × 2/day; Psyllogel(®) Fibra, Nathura, Montecchio Emilia, Italy) during the rehabilitative cycle. After a preliminary clinical evaluation, including the obstructed defecation syndrome (ODS) score, patients underwent defecography and anorectal manometry as well as rehabilitative treatment according to the "multimodal rehabilitative program" for obstructive defecation. At the end of the program, patients were reassessed by clinical evaluation and anorectal manometry. Post-rehabilitative ODS scores were used for an arbitrary schedule of patients divided into three classes: Class I, good (score ≤ 4); Class II, fair (score 4 to ≤ 8); Class III, poor (score 8).The number of bowel movements per week did not increase significantly after rehabilitation. Both groups had a significantly better Bristol stool form scale score (Group 1: P 0.034; Group 2: P 0.02). The overall mean ODS score from Groups 1 and 2 showed significant improvement after treatment (P 0.001). Twenty-eight patients (82.3%) were Class I (good results) without significant differences between groups. Nine women were symptom-free. Significant differences were found between pre-rehabilitative and post-rehabilitative manometric data from the straining test (P 0.001) and duration of maximal voluntary contraction (Group 1: P 0.004; Group 2: P 0.02). A significant difference was found between the pre-rehabilitative and post-rehabilitative conscious rectal sensitivity threshold (CRST) in Group 2 women (P 0.02). The Group 2 women who underwent volumetric rehabilitation (11 patients) had significantly lower post-rehabilitative CRST values than pre-rehabilitative values (P 0.002); the length of volumetric rehabilitation was also significantly shorter in Group 2 patients (P 0.04) than in Group 1 patients.After rehabilitation of obstructed defecation, some patients became symptom-free and many had an improved ODS score. Psyllium is helpful for volumetric rehabilitation: patients who consumed psyllium had lower post-rehabilitative CRST values than subjects were on high-fiber diet.

Background Fine-needle aspiration biopsy (FNAB) is the primary means to distinguish benign from malignant nodules and select patients for surgery. However, adjunctive diagnostic tests are needed because in 20-40% of cases the FNAB result is uncertain. Objective We investigated whether a search for the oncogenes RET/PTC, TRK and BRAF(V600E) in thyroid aspirates could refine an uncertain diagnosis. Patients and methods A total of 132 thyroid aspirates, including colloid nodules, inadequate samplings, indeterminate and suspicious for malignancy were analysed by reverse transcription polymerase chain reaction (RT-PCR) and mutant allele-specific amplification techniques for the presence of oncogenes. Results No oncogenes were detected in 48 colloid nodules, 46 inadequate and 19 indeterminate FNABs, then confirmed to be benign at histology. No oncogenes were detected in one follicular thyroid cancer (FTC) with indeterminate cytology. Five out of six papillary thyroid cancers (83%) with FNAB suspicious for malignancy were correctly diagnosed by the presence of oncogenes. Among these, four (67%) contained the BRAF mutation and one (17%) contained RET/PTC-3. On final analysis, no false-positive results were reported in 131 samples and five out of seven carcinomas (71%) were correctly diagnosed. The finding of oncogenes in FNAB specimens suspicious for malignancy guided the extent of surgical resection, changing the surgery from diagnostic to therapeutic in five cases. Conclusions Detection of RET/PTC, TRK and BRAF(V600E) in FNAB specimens is proposed as a diagnostic adjunctive tool in the evaluation of thyroid nodules with suspicious cytological findings.