Percussion

Adult male C57BL/6J mice (20–24 g) were subjected to midline fluid percussion injury (mFPI) consistent with methods previously described [27] . Animal numbers are indicated in the results section and figure legends for individual studies. Mice were anesthetized using 5% isoflurane in 100% oxygen for five minutes and the head of the animal was placed in a stereotaxic frame with continuously delivered isoflurane at 2.5% via nosecone. While anesthetized, the animal's body temperature was maintained using a Deltaphase® isothermal heating pad (Braintree Scientific Inc., Braintree, MA). A midline incision was made exposing bregma and lambda, and fascia was removed from the surface of the skull. A trephine (3 mm outer diameter) was used for the craniotomy, centered on the sagittal suture between bregma and lambda without disruption of the dura. An injury cap prepared from the female portion of a Luer-Loc needle hub was fixed over the craniotomy using cyanoacrylate gel and methyl-methacrylate (Hygenic Corp., Akron, OH). The incision was sutured at the anterior and posterior edges and topical Lidocaine ointment was applied. The injury cap was closed using a Luer-Loc cap and animals were placed in a heated recovery cage and monitored until ambulatory before being returned to their sleep cage.
For injury induction 24 hours post-surgery, animals were re-anesthetized with 5% isoflurane delivered for five minutes. The cap was removed from the injury-hub assembly and the craniotomy was visually inspected through the hub. The hub was then filled with normal saline and attached to the male end of the fluid percussion device (Custom Design and Fabrication, Virginia Commonwealth University, Richmond, VA). An injury of moderate severity (1.2–1.3 atm), mild severity (0.8 atm) or sham injury was administered by releasing the pendulum onto the fluid-filled cylinder. Sham-injured animals underwent the same procedure except the pendulum was not released. Animals were monitored for the presence of a forearm fencing response, and righting reflex times were recorded for the injured animals as indicators of injury severity [28] (link). The righting reflex time is the total time from the initial impact until the animal spontaneously rights itself. The fencing response is a tonic posturing characterized by extension and flexion of opposite arms that has been validated as an overt indicator of injury force magnitude [28] (link). The injury hub was removed and the brain was inspected for uniform herniation and integrity of the dura. Animals in which the dura was compromised were excluded from all studies as technical failures. The incision was cleaned using saline and closed using sutures. Moderate brain-injured animals had righting reflex recovery times greater than six minutes and a positive fencing response, and mild injured animals had righting reflex times between two and four minutes and no fencing response. Sham injured animals recovered within 10 seconds. After spontaneously righting, animals were placed in a heated recovery cage and monitored until ambulatory before being returned to their sleep cage (approximately 5 to 15 minutes). Adequate measures were taken to minimize pain or discomfort.

Adults (aged ≥18 years) presenting to the emergency department with a suspicion of CAP and fulfilling at least two of the following criteria: new or worsening cough, new or worsening expectoration of sputum, new or worsening dyspnoea, hemoptysis, pleuritic chest pain, radiological evidence of pneumonia, abnormalities on chest auscultation and/or percussion, and fever (≥38.0°C).
Written informed consent is needed from the patient or from their legal guardian/close relative at the time of recruitment.

This study was part of a project on neuroaesthetics, which addressed issues pertaining to domain-general and domain-specific neural organization among art students in a variety of fields (visual arts, dance, piano, strings, vocals, percussions) and non-artist healthy controls. To facilitate inter-group comparisons, we had all of the artists undergo the same psychological assessments and neuroimaging using the same scanning protocols. In the current study, we focused exclusively on VAs and controls. Creativity can be a core mental competence of VA since the process of creating artwork reflects VA’s creativity (Getzels and Csikszentmihalyi, 2020 ). Thus, all participants took the self-reported 40-item Chinese version of the Abbreviated Torrance Test for Adults (ATTA) to assess their aptitude in tasks involving visual (figural) and verbal manipulation (Chen, 2006 ). The ATTA is commonly used for cross-artist group comparisons (not addressed in the current study); however, it includes a figural part that engages creative drawing mirroring the visual art training of VAs. The ATTA measures the ability to think creatively in terms of fluency, originality, elaboration, and flexibility (Chen, 2006 ). Fluency refers to the number of ideas that a participant can generate in a limited time. Originality indicates one’s ability to create unique ideas. Elaboration indicates the ability to embellish ideas with details. Flexibility indicates one’s ability to generate many different ideas (Althuizen et al., 2010 (link); Shen and Lai, 2014 (link)). The ATTA creativity index (CI) score refers to the sum of the four capacity scores. We followed standard protocols in administering and scoring the tests (Chen, 2006 ). SPSS Statistics (v. 23.0, IBM Corp., Armonk, NY, USA) was used for all psychological evaluation analyses. The results of the psychological evaluations were considered significant at p < 0.05.

Study participants were three patients (two female and one male patient; mean age, 51 years) with chief complaint of severe lower anterior teeth mobility and severe periodontal compromission hindering any periodontal or conservative treatment. As control case, a patient (#1) referred with chief complaint due to lower third molar pericoronitis and indication for surgical extraction was recruited.
The exclusion criteria for this study were antibiotic therapy administered up to 3 months before tooth extraction, systemic diseases, and pregnancy. Another exclusion criterium was evaluating that periodontal pockets could not reach the apical root segment. Moreover, inclusion criteria were teeth affected by EPL without clinically and radiographically identified caries lesions, cracks/fractures and/or restorations.
The patients involved in the research signed a formal written informed consent form.
EPL diagnosis was performed with periodontal probing that evaluate an average clinical attachment loss ≥5 mm on all root’s surfaces and radiographic analyses on periapical bidimensional radiographies using paralleling technique confirming bone loss extending to mid-third of root and beyond; moreover, periapical radiolucency was not observed. None of these teeth presented periodontal pockets reaching the apex. Teeth presented Grade 2 mobility with percussion and palpation sensitivity. In addition, thermal and electric pulp sensibility tests were performed returning non-responsivity and thus confirming pulpar necrosis. Thermal pulp test was performed with a #2 cotton pellet sprayed and fully saturated with 1,1,1,2-tetrafluoroethane and placed at the middle third of the buccal tooth’s surface of the clinical crown for at least 20 s. Cold pulp testing was selected as pulp sensibility test which is able to evaluate vital (specificity = 0.84) and non-vital (sensitivity = 0.87) teeth [10–12 (link)]. No other EPL signs/symptoms (sinus tract, spontaneous pain, periapical radiolucency, suppuration) were observed.
Moreover, on intraoral inspection using loupes for magnification 4.0× and radiographic evaluation, the teeth did not present clinically identified caries lesions, cracks, fractures nor previous conservative restorations. The final diagnosis for all teeth was of EPL without root damage of Grade 3, according to the classification from the American Academy of Periodontology criteria. The sampled teeth clinically appeared without defects, decay, or restorations and were affected by severe periodontitis (Stage IV) according to the classification of American Academy of Periodontology criteria [5 (link)]. Control sample consisted of an intact lower third molar without carious and periodontal pathologies, surgically extracted because affected by pericoronitis. All teeth affected by EPL were single-rooted (N = 5 lower incisors and N = 1 lower canine).
A total of 12 clinical samples of the study group were collected from periodontal (P) (n = 6) and endodontic (E) (n = 6) tissue samples of root canals from six intact teeth of three patients (P1 and E1 from patient #1, P2 to P5 and E2 to E5 from patient #2, and P6 and E6 from patient #3).
All teeth were single-rooted (lower incisors and lower canine).
The study was performed in agreement with the ethical guidelines of the Declaration of Helsinki laid down in the 1964 and its later amendments or comparable ethical standards. The Ethics Committee of Azienda Unità Sanitaria Locale of Bologna approved this study with authorization no. 844-2021-OSS-AUSLBO-21160-ID 3118-Parere CE-AVEC-ENDO-MICROBIOTA 09/2021.

Experiments involving animals complied with ARRIVE guidelines and were performed in accordance with the National Institutes of Health guide for the care and use of laboratory animals (NIH publication No. 8023, revised 1978) following protocols approved by the University of California, Davis, Institutional Animal Care and Use Committee. Adult male Sprague Dawley rats (225–250 g, 8–10 weeks old) were purchased from Charles River Laboratories and individually housed in standard plastic cages under controlled environmental conditions (22 ± 2 °C, 40–50% humidity) in fully accredited AALAC International facilities. Animals were housed in a 12 h light-dark cycle with access to food and water provided ad libitum. Animals were acutely intoxicated with DFP (n = 10) as previously described [43 (link)]. Vehicle (VEH, n = 15) controls were administered saline instead of DFP but were otherwise similarly treated. Experimental TBI was produced using a fluid percussion device (VCU Biomedical Engineering [44 (link)]) with the lateral orientation for impact [45 (link)] as previously published (n = 5 [46 (link)]). Sham controls (n = 5) underwent the same surgical procedures but were not administered the percussive impact. All animal and group characteristics for each data set can be found in Table 1.Table 1

Animal distribution and group characteristics for exploratory and validation data sets.

Table 1

	Exploratory Data Set (Algorithm Development)		Validation Data Sets
Detection Method	Fluorescence		Fluorescence		Chromogen
Experimental Model	DFP Intoxication		DFP Intoxication		TBI
Brain Region Analyzed	Dorsal Lateral Thalamus		Dorsal Lateral Thalamus		Hippocampus
Final Group SizeDFP vs. VEHTBI vs. Sham	DFP:10	VEH:15	DFP:10	VEH:11	TBI:5	Sham:5
Primary AntibodySourceRRIDa	IBA-1Wako Laboratory ChemicalsAB_839504		IBA-1Wako Laboratory ChemicalsAB_839504		IBA-1Wako Laboratory ChemicalsAB_839504
	CD68Bio-Rad LaboratoriesAB_2291300		CD68Bio-Rad LaboratoriesAB_2291300

a

RRID: Research Resource Identification Number.