Maximal Fusion Capacity and Efficient Replenishment of the Dense Core Vesicle Pool in Hippocampal Neurons.

Neuropeptides and neurotrophins, stored in dense core vesicles (DCVs), are together the largest currently known group of chemical signals in the brain. Exocytosis of DCVs requires high-frequency or patterned stimulation, but the determinants to reach maximal fusion capacity and for efficient replenishment of released DCVs are unknown. Here, we systematically studied fusion of DCV with single vesicle resolution on different stimulation patterns in mammalian CNS neurons. We show that tetanic stimulation trains of 50-Hz action potential (AP) bursts maximized DCV fusion, with significantly fewer fusion event during later bursts of the train. This difference was omitted by introduction of interburst intervals but did not increase total DCV fusion. Interburst intervals as short as 5 s were sufficient to restore the fusion capacity. Theta burst stimulation (TBS) triggered less DCV fusion than tetanic stimulation, but a similar fusion efficiency per AP. Prepulse stimulation did not alter this. However, low-frequency stimulation (4 Hz) intermitted with fast ripple stimulation (200 APs at 200 Hz) produced substantial DCV fusion, albeit not as much as tetanic stimulation. Finally, individual fusion events had longer durations with more intense stimulation. These data indicate that trains of 50-Hz AP stimulation patterns triggered DCV exocytosis most efficiently and more intense stimulation promotes longer DCV fusion pore openings.SIGNIFICANCE STATEMENT Neuropeptides and neurotrophins modulate multiple regulatory functions of human body like reproduction, food intake or mood. They are packed into dense core vesicles (DCVs) that undergo calcium and action potential (AP) fusion with the plasma membrane. In order to study the fusion of DCVs in vitro, techniques like perfusion with buffer containing high concentration of potassium or electric field stimulation are needed to trigger the exocytosis of DCVs. Here, we studied the relationship between DCVs fusion properties and different electric field stimulation patterns. We used six different stimulation patterns and showed that trains of 50-Hz action potential bursts triggered DCV exocytosis most efficiently and more intense stimulation promotes longer DCV fusion pore openings.

Reduced dynamin-1 levels in neurons lacking MUNC18-1.

MUNC18-1 (also known as syntaxin-binding protein-1, encoded by Stxbp1) binds to syntaxin-1. Together, these proteins regulate synaptic vesicle exocytosis and have a separate role in neuronal viability. In Stxbp1 null mutant neurons, syntaxin-1 protein levels are reduced by 70%. Here, we show that dynamin-1 protein levels are reduced at least to the same extent, and transcript levels of Dnm1 (which encodes dynamin-1) are reduced by 50% in Stxbp1 null mutant brain. Several, but not all, other endocytic proteins were also found to be reduced, but to a lesser extent. The reduced dynamin-1 expression was not observed in SNAP25 null mutants or in double-null mutants of MUNC13-1 and -2 (also known as Unc13a and Unc13b, respectively), in which synaptic vesicle exocytosis is also blocked. Co-immunoprecipitation experiments demonstrated that dynamin-1 and MUNC18-1 do not bind directly. Furthermore, MUNC18-1 levels were unaltered in neurons lacking all three dynamin paralogues. Finally, overexpression of dynamin-1 was not sufficient to rescue neuronal viability in Stxbp1 null mutant neurons; thus, the reduction in dynamin-1 is not the single cause of neurodegeneration of these neurons. The reduction in levels of dynamin-1 protein and mRNA, as well as of other endocytosis proteins, in Stxbp1 null mutant neurons suggests that MUNC18-1 directly or indirectly controls expression of other presynaptic genes.

CaMKII controls neuromodulation via neuropeptide gene expression and axonal targeting of neuropeptide vesicles.

Ca2+/calmodulin-dependent kinase II (CaMKII) regulates synaptic plasticity in multiple ways, supposedly including the secretion of neuromodulators like brain-derived neurotrophic factor (BDNF). Here, we show that neuromodulator secretion is indeed reduced in mouse α- and ßCaMKII-deficient (αßCaMKII double-knockout [DKO]) hippocampal neurons. However, this was not due to reduced secretion efficiency or neuromodulator vesicle transport but to 40% reduced neuromodulator levels at synapses and 50% reduced delivery of new neuromodulator vesicles to axons. αßCaMKII depletion drastically reduced neuromodulator expression. Blocking BDNF secretion or BDNF scavenging in wild-type neurons produced a similar reduction. Reduced neuromodulator expression in αßCaMKII DKO neurons was restored by active ßCaMKII but not inactive ßCaMKII or αCaMKII, and by CaMKII downstream effectors that promote cAMP-response element binding protein (CREB) phosphorylation. These data indicate that CaMKII regulates neuromodulation in a feedback loop coupling neuromodulator secretion to ßCaMKII- and CREB-dependent neuromodulator expression and axonal targeting, but CaMKIIs are dispensable for the secretion process itself.

Patient-Specific Facial Implants in Polyetheretherketone and Their Stability: A Preliminary Study.

BACKGROUND: Polyetheretherketone (PEEK) is a synthetic material with many favorable characteristics; PEEK implants are increasingly used for a variety of applications ranging from cranioplasty to orthopedic surgery and facial implants. METHODS: This study is a retrospective review of patients who underwent PEEK implant placement in our department over the last 5 years. Polyetheretherketone computer-aided design and manufacture facial implants were designed from high-resolution computed tomography (CT) scans of each patient. The implants placed were onlay implants used for facial rehabilitation purposes to correct malformative and posttraumatic malformations. RESULTS: Twenty-eight consecutive patients (11 males and 17 females) underwent PEEK implant positioning between January 2015 and December 2020. Common indications were anterior plagiocephaly, hemifacial microsomia, and residual facial imbalance after orthognathic surgery. No complications of implant breakdown, exposure, infection, or displacement were noticed during the follow-up period. During routine controls on 3 patients, we requested a craniomaxillofacial CT scan for reasons unrelated to the implanted prostheses. The CT scans were all high resolution (<1-mm slices). The CT images indicated that bone was starting to form around the implant in all 3 patients as well as in the penetrating holes that were planned in the implants. CONCLUSIONS: In our experience, computer-designed, patient-specific PEEK onlay implants are a valid option for the treatment of malformative and posttraumatic malformations. This is, to the best of our knowledge, the first clinical report on bone reaction to PEEK implantation in the maxillofacial field. Moreover, based on the signs of bone regrowth that we observed in CT controls we can presume that the design of this type of prosthesis can probably take advantage of some technical stratagems not yet codified and fully exploited. Despite our preliminary favorable results, further multicentric and comparative studies are necessary to evaluate outcomes and better understand the behavior of this promising material and thus optimize its use in craniomaxillofacial surgery.

Pool size estimations for dense-core vesicles in mammalian CNS neurons.

Neuropeptides are essential signaling molecules transported and secreted by dense-core vesicles (DCVs), but the number of DCVs available for secretion, their subcellular distribution, and release probability are unknown. Here, we quantified DCV pool sizes in three types of mammalian CNS neurons in vitro and in vivo Super-resolution and electron microscopy reveal a total pool of 1,400-18,000 DCVs, correlating with neurite length. Excitatory hippocampal and inhibitory striatal neurons in vitro have a similar DCV density, and thalamo-cortical axons in vivo have a slightly higher density. Synapses contain on average two to three DCVs, at the periphery of synaptic vesicle clusters. DCVs distribute equally in axons and dendrites, but the vast majority (80%) of DCV fusion events occur at axons. The release probability of DCVs is 1-6%, depending on the stimulation. Thus, mammalian CNS neurons contain a large pool of DCVs of which only a small fraction can fuse, preferentially at axons.

Multidisciplinary ocular and periocular cancers meetings: implementation in a tertiary referral center and analysis over a 12-months period.

PURPOSE: The complexity of multimodal approaches in cancer management has lately led to the establishment of multidisciplinary tumor boards (MDTBs) to define targeted, patient-centered treatment strategies. However, few data are available regarding the application of this approach in Ocular Oncology. Hereby, the Authors analyze the implementation and outcomes of a trained MDTB in a tertiary ocular oncology referral center. METHODS: A retrospective descriptive analysis of MDTB meetings discussing patients with ocular and periocular cancers, over a 12-months period, was carried out. Data were grouped by main site involved, topics discussed and final clinical decisions therefore taken. Meetings were held by a constant 'Core team' or - when required - by a broader 'Extended team'. RESULTS: During the observational period 86 cases were discussed. In 27 patients ocular surface tissues were involved (31%), in 25 patients orbital tissues (29%), in 22 patients eyelids (26%), and in 12 patients intraocular tissues (14%). In 13 cases (15%) naïve or referred new patients, in 34 cases (40%) imaging or histopathologic reports and in 39 cases (45%) treatment plans were discussed. Regarding final decisions, a treatment plan was scheduled in 47 cases (55%) and a diagnostic ascertainment was required in 27 patients (31%); locally advanced and/or systemic diseases were referred or teamed up with other specialists in 12 cases (14%). CONCLUSIONS: Ocular Oncology multidisciplinary team, by sharing expertise of different specialists, ensures a comprehensive evaluation of patients improving the accuracy of diagnosis and staging upon which planning a proper treatment. Further studies are needed to assess if this approach may also improve the outcomes and prognosis of patients.

Patient First: Surgery First and More.

INTRODUCTION: The success of surgery first approach has been defined by the number of advantages offered and is definitively determined by the satisfaction of the patients themselves. The strength of this protocol resides in its philosophy that puts the patient at the center of the whole diagnostic-therapeutic process. The compliance of the patient, its happiness and comfort are the best guarantee of a good final results. While pursuing this philosophy we have wondered whether there was way to increase the comfort of surgery first approach even more and to make it even more appealing for the patients. For these reasons, we have decided to work on a preliminary protocol in order to reduce or even eliminate the use of orthodontic braces and wires during perioperative stages. Materials and Methods: No orthodontic braces or wires are bonded on the teeth before surgery. Intraoperatory intermaxillary fixation (IMF) is carried out with the use of IMF screws which are positioned at the beginning of the operation on the edge between keratinized and nonkeratinized gingiva. Once the osteotomies are performed IMF with IMF screws is carried out on the planned occlusion with the use of surgical splints.Discussion and Conclusions: In selected cases the postsurgical orthodontic treatment can be carried out with the use of clear aligners, completely eliminating the need of braces and wires with an additional level of comfort.

Approach to Fronto-Orbital Sequelae in Anterior Synostotic Plagiocephaly: Our Flow Chart Based on Di Rocco Classification.

INTRODUCTION: Anterior synostotic plagiocephaly recognizes the synostosis of one of the hemicoronal sutures as a cause and can manifest itself with varying degrees of severity. Clinically it presents a reduction of the sagittal growth of the affected side and flattening of the frontoparietal complex. MATERIALS AND METHODS: The authors retrospectively examined our case sample dividing it into 3 groups based on the Di Rocco classification. For each category, we assessed the extent of facial alterations at the end of skeletal growth and retrospectively analyzed the surgical options aimed at correcting aesthetic and skeletal deficits. RESULTS: The authors found that predictable results could be obtained by standardizing the surgical procedure based on Di Rocco's classification groups; in particular, the authors achieved satisfactory results by assigning a specific surgical procedure to each class.

Image Captioning Using Motion-CNN with Object Detection.

Automatic image captioning has many important applications, such as the depiction of visual contents for visually impaired people or the indexing of images on the internet. Recently, deep learning-based image captioning models have been researched extensively. For caption generation, they learn the relation between image features and words included in the captions. However, image features might not be relevant for certain words such as verbs. Therefore, our earlier reported method included the use of motion features along with image features for generating captions including verbs. However, all the motion features were used. Since not all motion features contributed positively to the captioning process, unnecessary motion features decreased the captioning accuracy. As described herein, we use experiments with motion features for thorough analysis of the reasons for the decline in accuracy. We propose a novel, end-to-end trainable method for image caption generation that alleviates the decreased accuracy of caption generation. Our proposed model was evaluated using three datasets: MSR-VTT2016-Image, MSCOCO, and several copyright-free images. Results demonstrate that our proposed method improves caption generation performance.

Virtual Surgical Planning for Reconstruction of Giant Ameloblastoma of the Mandible.

Ameloblastoma is a histologically benign tumor that behaves aggressively because of its tendency to invade local structures, and it has a high probability of local recurrence. If neglected, ameloblastomas can grow substantially over the course of years, reaching the size of giant ameloblastomas. This large size can lead to deformities in facial appearance and impairments in speaking, swallowing, eating, and breathing.Surgical planning can be challenging because of the extension of the tumor and the consequent reconstructive issues.In this article, we present our experience with the reconstruction of 2 cases of giant ameloblastomas planned on the basis of occlusal casts and acrylic splints. In these patients, computerized planning was rendered complex and potentially inaccurate because of the dimensions of the tumor, the loss of anatomical landmarks, and the loss of occlusal landmarks. The cases were successfully reconstructed, but the technique can be flawed. A 3-dimensional virtual model of the mandible can be used as a template to develop cutting guides for reconstruction with free fibular flaps. This will allow us to overcome limitations, standardize the procedure, and achieve optimal functional and aesthetic results.

Robust and Efficient Indoor Localization Using Sparse Semantic Information from a Spherical Camera.

Self-localization enables a system to navigate and interact with its environment. In this study, we propose a novel sparse semantic self-localization approach for robust and efficient indoor localization. "Sparse semantic" refers to the detection of sparsely distributed objects such as doors and windows. We use sparse semantic information to self-localize on a human-readable 2D annotated map in the sensor model. Thus, compared to previous works using point clouds or other dense and large data structures, our work uses a small amount of sparse semantic information, which efficiently reduces uncertainty in real-time localization. Unlike complex 3D constructions, the annotated map required by our method can be easily prepared by marking the approximate centers of the annotated objects on a 2D map. Our approach is robust to the partial obstruction of views and geometrical errors on the map. The localization is performed using low-cost lightweight sensors, an inertial measurement unit and a spherical camera. We conducted experiments to show the feasibility and robustness of our approach.

Value of the Rare Disease Registry of the Italian Region Friuli Venezia Giulia.

BACKGROUND: The lack of epidemiological and clinical data is a major obstacle in health service planning for rare diseases. Patient registries are examples of real-world data that may fill the information gap. OBJECTIVE: We describe the Rare Disease Registry of the Friuli Venezia Giulia region of Italy and its potential for research and health planning. METHODS: The Rare Disease Registry data were linked with information on mortality, hospital discharges, ambulatory care, and drug prescriptions contained in administrative databases. All information is anonymous, and data linkage was based on a stochastic key univocal for each patient. Average annual costs owing to hospitalizations, outpatient care, and medications were estimated. RESULTS: Implementation of the Registry started in 2010, and 4250 participants were registered up to 2017. A total of 2696 patients were living in the region as of January 1, 2017. The overall raw prevalence of rare diseases was 22 per 10,000 inhabitants, with higher prevalence in the pediatric population. The most common disease groups were congenital malformations, chromosomal and genetic syndromes, and circulatory and nervous diseases. In 2017, 30 patients died, 648 were hospitalized, and 2355 received some type of ambulatory care. The total annual estimated cost was approximately €6.5 million, with great variability in the average patient cost across diseases. CONCLUSIONS: The possibility of following the detailed real-world care experience of patients with each specific rare disease and assessing the costs related to each step in their care path represents a unique opportunity to identify inefficiencies, optimize care, and reduce waste of resources.

Use of Navigation System in Craniofacial Surgery.

The use of navigated surgery in the treatment of craniofacial malformations can help obtain optimal results. In this article, the authors will discuss a case of anterior plagiocephaly, corrected with frontorbital bandeau remodeling. Navigation was used during the osteotomy and the reposition phase to ensure the correct positioning of the osteotomy instruments. It was also used to ensure that the bandeau was correctly repositioned in accordance with the surgical plan determined during the virtual simulation phase of the surgery.

Early Surgical Treatment in Anterior Synostotic Plagiocephaly: Is This the Best Choice?

At the end of craniofacial growth, in anterior synostotic plagiocephaly, some aesthetical deficiencies may still be evident. This can depend on an inadequate initial correction or on altered postoperative growth or even on the combination of the 2 factors.Aesthetic alterations can result from various factors that could potentially affect the skeleton, the skin, subcutaneous, and muscular tissues.The pathological changes in the cutaneous and subcutaneous tissues are greater in patients who have undergone multiple surgical treatments of the frontoorbital area. The aim of this observational cohort study is to assess the residual aesthetic and functional impairment at the end of skeletal growth, in patients affected by anterior synostotic plagiocephaly who have undergone surgery at an early age. The purpose is to investigate whether early surgery can still be considered unavoidable in patients with this malformation.Between July 2012 and February 2015, patient's data were retrieved from our archives among the patients referred to our department from 2003 to 2012 for Anterior Synostotic Plagiocephaly at an early age.The authors studied this patient with CT scans and photographic documentation. On CT scans, the authors have assessed skeletal alterations, soft tissues alterations, and muscular tissue alterations. With photographic documentation, the authors have studied the perception of the malformation among external subjects.From this study it was possible to demonstrate that is many esthetical alterations are to still to be found in patients treated with an early surgical approach; for this reason in children without early complications, the authors suggest that surgical treatment should be delayed after the end of craniofacial growth.

Effect of Surgery First Orthognathic Approach on the Temporomandibular Joint.

INTRODUCTION: Correction of severe malocclusions with skeletal discrepancies requires orthodontic treatment in combination with orthognathic surgery. Even though conventional orthognathic surgery (COS) is a common and well-accepted approach its influence on the signs and symptoms of temporomandibular disorders (TMDs) is still debated. Recently with the introduction of surgery first approach, a different timing for the management of dentoskeletal imbalances has been proposed. The present study is aimed at assessing the relationship between surgery first approach and temporomandibular joint (TMJ) disorders. METHODS: The study sample consisted of 24 patients who were selected to be treated with surgery first approach. Clinical follow-ups after surgery were performed every week for the first month, at 3 months, 6 months, and at 1 year. A radiological follow-up was performed at 1 week and at 1 year after the operation with a panorex and a latero-lateral teleradiograph. To assess the effect of surgery first approach on the TMDs signs and symptoms, a clinical assessment was performed 4 days before surgery (T1), 6 months after surgery (T2), and 1 year postoperatively (T3). RESULTS: The results of the authors' study show that pain assessment revealed a general improvement of this symptom in correspondence to TMJ and masticatory muscles except in the masseter and neck region. Also joint noises, TMJ functioning, migraine, and headache underwent a considerable improvement. CONCLUSION: Surgery first approach is an innovative orthognathic procedure and, by undergoing surgery first approach, patients with pre-existing TMJ dysfunction may experience a significant improvement or even resolution of the TMDs signs and symptoms.

The Arterialized Facial Artery Musculo-Mucosal Island Flap for Post-Oncological Tongue Reconstruction.

BACKGROUND: In 1992, Pribaz described the facial artery musculomucosal flap (FAMM), an axial musculomucosal flap based on the facial artery. The FAMM flap, a modification of the nasolabial and buccal mucosal flaps, is widely used in the reconstruction of defects in the oral cavity. Many modifications of this flap have been described in the literature. Here we aimed to explore the use of an arterialized tunnelized FAMM island flap (a-FAMMIF) for the reconstruction tongue defects after tumor resection. METHOD: From January 2015 to December 2016, five cases of tongue cancer were selected for the use of arterialized FAMMIF flap to reconstruct defects after tumor resection. RESULTS: Reconstruction was successful in all cases, except one case of total flap necrosis; partial necrosis of the flap occurred in two patients, which were solved with medications. CONCLUSION: The authors consider the a-FAMMIF an unreliable flap in the reconstruction of tongue defects.The authors recommend avoiding tunneling and island modification when the vein is not included in the pedicle.

The Rules of Attractiveness: A Study on the Lower Facial Third.

What is considered attractive may not fall into the "norm," and it can vary from culture to culture and depending on the historical time, for this reason the standard cephalometric and antropometric references may not be sufficient in these cases.Lately some techniques have arose to popularity that are aimed to changing the frontal and lateral aspect of the facial lower third, such as V-line or the Chin-Wing Osteotomy technique, but no reference system exists at the moment to define to which extent a modification of the lower third falls within what is considered beautiful, and everything is left to the patient's will or to the surgeon's sensitivity.The aim of this article is to study which antropometric value is considered attractive by the most for what concerns the frontal shape of the lower third of the face.Twenty-four female models were enrolled in this study and the angle taken into consideration was the one at the intersection between the 2 lines connecting the cutaneous gonial angle of each side of the face and the most external part of the chin on the same side. Measures were made on pictures in frontal view.Two hundred two random examiners were asked to see the pictures and rate them as attractive or nonattractive.Results were then paired with the angles values.Among the models the higher angle measured was 107.5° (found in 1 individual) while the lower angle was 76° (found in 1 individual), the average measure calculated was 88.3° while the median angle was 89.5°.According to the result the subjects considered more attractive were those with an angle between 84.5 and 91.5 (92 for male examiners).This could be an important starting point for studies who can evaluate attractiveness from a numerical point of view.

Navigation System Approach in Zygomatic Implant Technique.

In some patients, the resolution of severe maxillary atrophies can be hardly achieved without the use of zygomatic implants. Although many scientific studies have already demonstrated the excellent immediate stability in long term, the use of zygomatic implants is not yet widespread. Among the complications of this technique, the most threatening is the risk of damaging the eyeball or the maxillary nerve.The use of the navigator system as a surgical aid for implant placement allows to control, at any time, the position of the drill in the bone, avoiding any injury to ocular and nervous structures. The authors present a clinical report which shows a patient affected by a very severe form of post-traumatic maxillary atrophy that has been solved through the of zygomatic implants placement using the "Implant Bone Navigation" system. This procedure allows both to cut down the risks on ocular and nervous structures of the maxilla and also to reach excellent rehabilitation results in such severely compromised patients.

Surgery-first orthognathic approach vs traditional orthognathic approach: Oral health-related quality of life assessed with 2 questionnaires.

INTRODUCTION: The purposes of the study were to investigate and evaluate the differences detected by the patients between the traditional orthognathic approach and the surgery-first one in terms of level of satisfaction and quality of life. METHODS: A total of 30 patients who underwent orthognathic surgery for correction of malocclusions were selected and included in this study. Fifteen patients were treated with the conventional orthognathic surgery approach, and 15 patients with the surgery-first approach. Variables were assessed through the Orthognathic Quality of Life Questionnaire and the Oral Health Impact Profile questionnaire and analyzed with 2-way repeated-measures analysis of variance. RESULTS: The results showed significant differences in terms of the Orthognathic Quality of Life Questionnaire (P <0.001) and the Oral Health Impact Profile (P <0.001) scores within groups between the first and last administrations of both questionnaires. Differences in the control group between first and second administrations were also significant. Questionnaire scores showed an immediate increase of quality of life after surgery in the surgery-first group and an initial worsening during orthodontic treatment in the traditional approach group followed by postoperative improvement. CONCLUSIONS: This study showed that the worsening of the facial profile during the traditional orthognathic surgery approach decompensation phase has a negative impact on the perception of patients' quality of life. Surgeons should consider the possibility of a surgery-first approach to prevent this occurrence.

Trancutaneous Versus Intraoral Approach to Isolated Zygomatic Arch Fractures: A Comparison of Two Techniques.

Zygomatic arch fractures are caused by a vector force orthogonal to the bone segment that causes the collapse of the arch through depression of the bone fragments. Reduction of isolated zygomatic arch fractures are usually only of esthetic interest, with the exception of those cases where the fracture causes an impingement with the underlying mandibular coronoid process, causing limitation of mandibular movements. Reduction is usually performed with an extraoral approach, more rarely through a transoral approach. In this article, authors compare the traditional transcutaneous technique with the intraoral approach in 2 groups for a total number of 42 patients.For what concerns the correct alignment of the fragments, the 2 techniques have shown being equivalent. Although the intraoral approach has shown being a faster surgical procedure leaving no visible incision, allowing faster recovering and reduced postoperative pain.