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Behavioral testing is an essential tool for evaluating cognitive function and dysfunction in preclinical research models. This is of special importance in the study of neurological disorders such as Alzheimer's disease. However, the reproducibility of classic behavioral assays is frequently compromised by interstudy variation, leading to ambiguous conclusions about the behavioral markers characterizing the disease. Here, we identify age- and genotype-driven differences between 3xTg-AD and non-transgenic control mice using a low-cost, highly customizable behavioral assay that requires little human intervention. Through behavioral phenotyping combining both supervised and unsupervised behavioral classification methods, we are able to validate the preventative effects of the immunosuppressant cyclosporine A in a rodent model of Alzheimer's disease, as well as the partially ameliorating effects of candidate drugs nebivolol and cabozantinib.

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Airway integrity must be continuously maintained throughout life. Sensory neurons guard against airway obstruction and, on a moment-by-moment basis, enact vital reflexes to maintain respiratory function1,2. Decreased lung capacity is common and life-threatening across many respiratory diseases, and lung collapse can be acutely evoked by chest wall trauma, pneumothorax or airway compression. Here we characterize a neuronal reflex of the vagus nerve evoked by airway closure that leads to gasping. In vivo vagal ganglion imaging revealed dedicated sensory neurons that detect airway compression but not airway stretch. Vagal neurons expressing PVALB mediate airway closure responses and innervate clusters of lung epithelial cells called neuroepithelial bodies (NEBs). Stimulating NEBs or vagal PVALB neurons evoked gasping in the absence of airway threats, whereas ablating NEBs or vagal PVALB neurons eliminated gasping in response to airway closure. Single-cell RNA sequencing revealed that NEBs uniformly express the mechanoreceptor PIEZO2, and targeted knockout of Piezo2 in NEBs eliminated responses to airway closure. NEBs were dispensable for the Hering-Breuer inspiratory reflex, which indicated that discrete terminal structures detect airway closure and inflation. Similar to the involvement of Merkel cells in touch sensation3,4, NEBs are PIEZO2-expressing epithelial cells and, moreover, are crucial for an aspect of lung mechanosensation. These findings expand our understanding of neuronal diversity in the airways and reveal a dedicated vagal pathway that detects airway closure to help preserve respiratory function.

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Introduction Osteoid osteomas are the most frequent true benign bone tumor in the adolescent age group and the third most prevalent benign bone tumor overall. This study was designed to assess the effectiveness of the procedure and correlate it with the analgesia offered because of the significant burden of this illness and new literature supporting the successful outcomes of image-guided percutaneous radiofrequency ablation (RFA) in osteoid osteoma. Methodology  This hospital-based interventional trial was carried out in a tertiary care referral center. Forty-two patients with osteoid osteoma, ranging in age from 9 to 30, were included in the study. The patients received RFA guided by computed tomography (CT), and they were postoperatively monitored at one, two, and four weeks and three, six months, and 12 months. A numerical pain scale (NPS) was used to evaluate the patient's pain both before and after the procedure. The preoperative and postoperative results were contrasted. Results A total of 42 participants were enrolled in the study. Eight (19.05%) women and 34 (80.95%) men made up the group. Complete pain alleviation (NPS=0) was attained in 42.8% and 96.4% of the study group in the first and second weeks post-procedure. Almost all patients began protected weight-bearing at one week, according to their level of pain tolerance. Osteoid osteoma of the talus was a remnant lesion in one patient that required further treatment after two weeks. During the duration of the follow-up, no problems were recorded. Conclusion Percutaneous CT-guided RFA of osteoid osteoma is a safe, minimally invasive procedure and greatly reduces the duration of hospitalization. It has excellent functional outcomes and no known complications.

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The analysis of mouse behavior is used in biomedical research to study brain function in health and disease. Well-established rapid assays allow for high-throughput analyses of behavior but have several drawbacks, including measurements of daytime behaviors in nocturnal animals, effects of animal handling, and the lack of an acclimation period in the testing apparatus. We developed a novel 8-cage imaging system, with animated visual stimuli, for automated analyses of mouse behavior in 22-h overnight recordings. Software for image analysis was developed in two open-source programs, ImageJ and DeepLabCut. The imaging system was tested using 4-5 month-old female wild-type mice and 3xTg-AD mice, a widely-used model to study Alzheimer's disease (AD). The overnight recordings provided measurements of multiple behaviors including acclimation to the novel cage environment, day and nighttime activity, stretch-attend postures, location in various cage areas, and habituation to animated visual stimuli. The behavioral profiles were different in wild-type and 3xTg-AD mice. AD-model mice displayed reduced acclimation to the novel cage environment, were hyperactive during the first hour of darkness, and spent less time at home in comparison to wild-type mice. We propose that the imaging system may be used to study various neurological and neurodegenerative disorders, including Alzheimer's disease.

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Neuronal up and down states have long been known to exist both in vitro and in vivo. A variety of functions and mechanisms have been proposed for their generation, but there has not been a clear connection between the functions and mechanisms. We explore the potential contribution of cellular-level biochemistry to the network-level mechanisms thought to underlie the generation of up and down states. We develop a neurochemical model of a single tripartite synapse, assumed to be within a network of similar tripartite synapses, to investigate possible function-mechanism links for the appearance of up and down states. We characterize the behavior of our model in different regions of parameter space and show that resource limitation at the tripartite synapse affects its ability to faithfully transmit input signals, leading to extinction-down states. Recovery of resources allows for "reignition" into up states. The tripartite synapse exhibits distinctive "regimes" of operation depending on whether ATP, neurotransmitter (glutamate), both, or neither, is limiting. Our model qualitatively matches the behavior of six disparate experimental systems, including both in vitro and in vivo models, without changing any model parameters except those related to the experimental conditions. We also explore the effects of varying different critical parameters within the model. Here we show that availability of energy, represented by ATP, and glutamate for neurotransmission at the cellular level are intimately related, and are capable of promoting state transitions at the network level as ignition and extinction phenomena. Our model is complementary to existing models of neuronal up and down states in that it focuses on cellular-level dynamics while still retaining essential network-level processes. Our model predicts the existence of a "final common pathway" of behavior at the tripartite synapse arising from scarcity of resources and may explain use dependence in the phenomenon of "local sleep." Ultimately, sleeplike behavior may be a fundamental property of networks of tripartite synapses.

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Enteroendocrine cells are specialized sensory cells of the gut-brain axis that are sparsely distributed along the intestinal epithelium. The functions of enteroendocrine cells have classically been inferred by the gut hormones they release. However, individual enteroendocrine cells typically produce multiple, sometimes apparently opposing, gut hormones in combination, and some gut hormones are also produced elsewhere in the body. Here, we developed approaches involving intersectional genetics to enable selective access to enteroendocrine cells in vivo in mice. We targeted FlpO expression to the endogenous Villin1 locus (in Vil1-p2a-FlpO knock-in mice) to restrict reporter expression to intestinal epithelium. Combined use of Cre and Flp alleles effectively targeted major transcriptome-defined enteroendocrine cell lineages that produce serotonin, glucagon-like peptide 1, cholecystokinin, somatostatin, or glucose-dependent insulinotropic polypeptide. Chemogenetic activation of different enteroendocrine cell types variably impacted feeding behavior and gut motility. Defining the physiological roles of different enteroendocrine cell types provides an essential framework for understanding sensory biology of the intestine.

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BACKGROUND: Simultaneous traumatic dislocation of both the hips in an individual is an uncommon injury. It accounts for only 1-2% of all traumatic hip dislocations. Simultaneous traumatic dislocation of both the hips in opposite directions i.e., anterior dislocation of one hip and posterior of the other is even rarer. These asymmetric dislocations need prompt identification for appropriate management. While their management should proceed in a manner commensurate with the accepted management of isolated dislocations to avoid complications, it is the timely identification which is essential. STUDY DETAILS: Retrospectively the details of three patients with asymmetric bilateral hip dislocations were collected. The article describes the clinical presentation of each patient and the management offered to them. The follow up duration ranged from 15 months to 24 months. RESULTS: In the short term, the outcomes in terms of hip pain, movements at the hips and radiological parameters like absence of arthritic or avascular changes were found to be satisfactory in each patient. CONCLUSION: Early diagnosis and prompt reduction is the essential first step in the appropriate management of these patients to prevent the possible complications. The associated fractures must be managed as per the established principles of trauma care. Long term follow up is essential for identifying if complications like avascular necrosis and secondary osteoarthritis develop after this uncommon injury.

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The convergence of internal path integration and external sensory landmarks generates a cognitive spatial map in the hippocampus. We studied how localized odor cues are recognized as landmarks by recording the activity of neurons in CA1 during a virtual navigation task. We found that odor cues enriched place cell representations, dramatically improving navigation. Presentation of the same odor at different locations generated distinct place cell representations. An odor cue at a proximal location enhanced the local place cell density and also led to the formation of place cells beyond the cue. This resulted in the recognition of a second, more distal odor cue as a distinct landmark, suggesting an iterative mechanism for extending spatial representations into unknown territory. Our results establish that odors can serve as landmarks, motivating a model in which path integration and odor landmarks interact sequentially and iteratively to generate cognitive spatial maps over long distances.

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The care of a patient with a spinal cord injury is part of healthcare systems. It causes a substantial physical and emotional drain on the caretakers who often are in short supply and thus may lack adequate training, preparation, and support. Long hours of assisting a chronically handicapped patient with activities of daily living and exercises decrease the rehabilitator's quality of life and take a psychological toll that increases a risk of burnout syndrome. The present study found a significant caregiving burden among care providers of chronically dependent patients with spinal cord injuries. Additionally, financial drain escalates the issue in this rather neglected health and quality of life aspect concerning caregivers. For the situation to improve, there must be a paradigm shift in care taking toward the motivative patient's participation in the rehabilitative process. Provisions for social support and educational programs focusing on the patients and their families need to be reappraised.

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Mechanosensory neurons across physiological systems sense force using diverse terminal morphologies. Arterial baroreceptors are sensory neurons that monitor blood pressure for real-time stabilization of cardiovascular output. Various aortic sensory terminals have been described, but those that sense blood pressure are unclear because of a lack of selective genetic tools. Here, we find that all baroreceptor neurons are marked in Piezo2-ires-Cre mice and then use genetic approaches to visualize the architecture of mechanosensory endings. Cre-guided ablation of vagal and glossopharyngeal PIEZO2 neurons eliminates the baroreceptor reflex and aortic depressor nerve effects on blood pressure and heart rate. Genetic mapping reveals that PIEZO2 neurons form a distinctive mechanosensory structure: macroscopic claws that surround the aortic arch and exude fine end-net endings. Other arterial sensory neurons that form flower-spray terminals are dispensable for baroreception. Together, these findings provide structural insights into how blood pressure is sensed in the aortic vessel wall.

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INTRODUCTION: Open comminuted distal femur fractures are notorious for septic or aseptic non-union. The recommended fixed angle distal femur locking plate in such situations can lead to a septic non-union due to its extensive approach and further periosteal stripping. Supracondylar nails, though have a minimally invasive approach, are not suitable for type C2 and C3 (AO/ASIF) fractures. A monolateral fixator as damage control followed by plating may be recommended. But if wound healing is delayed it results in difficult articular reduction, poor alignment and a stiff knee. We therefore used ilizarov circular external fixators (ICEF) for such open fractures (type C1, C2 and C3) and analysed its radiological and functional outcomes. MATERIALS AND METHODS: 25 male patients, with a mean age of 31.04 ±â€¯6.62 years (range, 22-44 years), with open grade III type C distal femoral fractures were treated with ICEF. There were 7 fractures of type C1and C3 each, 11 were of type C2. Articular reduction and compression was achieved with inter-fragmentary screws through minimal open technique by extending the open wound and then stabilising the fracture with ICEF. The main outcomes evaluated were union, range of motion, final shortening, Knee Society scoring and ASAMI scoring system for radiological and functional outcomes. RESULTS: The mean follow-up period was 19.12 ±â€¯1.14 months. All fractures except two united at a mean period of 30 ±â€¯3.02 weeks, without the need of bone grafts. The bony assessment (according to ASAMI score) was excellent in 8 cases (33.33%), good in 9 cases (37.5%) and fair in 5 cases (20.83%), while there were 2 poor clinical end results. The functional results were excellent in 6 cases (25%), good in 9 cases (37.5%) fair in 6 cases (25%) poor in 3 cases (12.5%). The complications included shortening, extension lag and pin tract infections. CONCLUSION: With the encouraging results, the use of ICEF with minimal internal fixation in grade III open comminuted distal femur fractures as a primary definitive treatment is a valuable alternative.

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BACKGROUND: Tibia fracture is the most common long bone fracture. The fractures of tibia are commonly open fractures due to subcutaneous position of the tibia. The choice of technique for stabilization of open tibia fractures includes - External fixation, unreamed intra-medullary nails [URTN], Reamed intra-medullary nails, ORIF with Plating. OBJECTIVES: To evaluate & compare the results of Unreamed Intra-Medullary Nail Versus Half Pin External Fixator in Grade III [A & B] Open tibia fractures. METHODS: This prospective clinical study [Randomized chit box] was done on 50 patients presenting to our institute within 24 h of injury. Only those who were skeletally mature with open tibia fracture Grade IIIA & IIIB [Gustilo-Anderson] were included in this study. After initial management, radiological assessment was done. Following this adequate wound debridement, skeletal stabilization with either primary URTN or external fixator was done. Inspection and debridement were repeated at 48-h intervals until the wound was considered clean. RESULTS: 50 cases [25 each group] were compared in terms of - Final Alignment of the Fracture, Presence of Infection/Non-union/Mal-union, Hardware failure, Time to Bone Union, Number of Operative Procedures after index admission. Mean time to full weight bearing was 20.96 weeks in URTN group versus 24.8 weeks in Ex-fix group. 5 in URTN group required further surgery for non-union versus 11 patients in Ex-fix group. There were 6 significant pin track infection. Removal of nail was required in 1 case of deep infection. CONCLUSION: This study supports the use of the URTN over External fixator in the treatment of severe open tibia fractures.

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The sense of taste allows animals to sample chemicals in the environment prior to ingestion. Of the five basic tastes, sour, the taste of acids, had remained among the most mysterious. Acids are detected by type III taste receptor cells (TRCs), located in taste buds across the tongue and palate epithelium. The first step in sour taste transduction is believed to be entry of protons into the cell cytosol, which leads to cytosolic acidification and the generation of action potentials. The proton-selective ion channel Otop1 is expressed in type III TRCs and is a candidate sour receptor. Here, we tested the contribution of Otop1 to taste cell and gustatory nerve responses to acids in mice in which Otop1 was genetically inactivated (Otop1-KO mice). We first show that Otop1 is required for the inward proton current in type III TRCs from different parts of the tongue that are otherwise molecularly heterogeneous. We next show that in type III TRCs from Otop1-KO mice, intracellular pH does not track with extracellular pH and that moderately acidic stimuli do not elicit trains of action potentials, as they do in type III TRCs from wild-type mice. Moreover, gustatory nerve responses in Otop1-KO mice were severely and selectively attenuated for acidic stimuli, including citric acid and HCl. These results establish that the Otop1 proton channel plays a critical role in acid detection in the mouse gustatory system, evidence that it is a bona fide sour taste receptor.

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INTRODUCTION: Giant-cell tumors (GCTs) are benign tumors of musculoskeletal system, commonly occur around knee. High-grade GCTs have a high tendency to recur after intralesional curettage. Reconstruction of joint after excision of such aggressive juxta-articular GCT has been challenging task. We describe a case of recurrence in GCT of lateral femoral condyle managed with tumor excision and pedicled patellar transplant to reconstruct the joint. CASE REPORT: A 25-year-old male patient reported to us after 22 months after he had undergone curettage and bone grafting for GCT of lateral femoral condyle of his right knee with recurrence of primary tumor. After tumor resection, we restored the lost articular surface with musculovascular pedicled patella and managed dead space with corticocancellous bone graft and fibular strut. 2-year follow-up exhibited good functional outcome without any sign of recurrence. CONCLUSION: Biological reconstruction with patellar transplant can be a good option for young patients in whom endoprosthesis placement poses significant concerns about longevity, cost, and revision surgeries.

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Recent studies have revealed that acrolein, a commonly found toxin and a potent metabolite of cyclophosphamide (CTX), can cause deterioration of mouse oocyte quality through a mechanism involving the generation of reactive oxygen species (ROS). We extend these studies to evaluate the effects of acrolein, in varying concentrations, on the oocyte mitochondrial membrane and oocyte apoptosis and its effect on embryo development in vitro. Metaphase II mouse oocytes were exposed for 45 minutes to acrolein and CTX (10 & 25 µM) and mitochondrial dysfunction, a major source of ROS overproduction, was evaluated by the 5,5,6,6-tetrachloro-1,1,3,3-tetraethyl-ß-benzimidazolylcarbocyanine iodide (JC-10) mitochondrial membrane potential assay. Treatment with acrolein led to mitochondrial membrane damage as well as induction of apoptosis compared to untreated control (p < 0.05). Similar results were obtained when oocytes were exposed to CTX (p < .05). Subsequently, the effect of acrolein exposure was evaluated by observing in vitro development of embryos after exposure. Acrolein treatment caused higher proportions of arrested and poor-quality embryos, evidenced by irregular cleavage, severe asymmetry of blastomeres, presence of large percentage of anuclear fragments, and dark granularity of the cytoplasm. Development at various durations in culture revealed that optimal embryo growth was significantly inhibited in a dose dependent manner, when compared to control (p < .05). A global model that links acrolein accumulation, generation of ROS, and mitochondrial dysfunction and their effect on oocyte and embryo quality is discussed further. Collectively, understanding the mechanism by which CTX and acrolein impact fertility is helpful in finding potential alternative or supplemental treatment options.

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BACKGROUND: Untreated Perthes disease can lead to osteoarthritis by the fourth decade. The treatment is conservative for children <6 years, operative within the age group of 6-9 years. Late onset Perthes, older than 9 years or more, are notorious with the aggressive course with poor outcome. However, literature do not come to a consensus between conservative and operative management. This study evaluates the clinical and radiological outcome of varus derotation osteotomy (VDRO) in Perthes disease presenting late with age 8 years or more. MATERIALS AND METHODS: 15 children (13 males and 2 females) with the mean age of 9.4 years belonging to modified Elizabethtown classification Stage IB, IIA, IIB treated with open wedge VDRO between 2008 and 2014 were included in this study. Seven patients (46.67%) were of >10 years of age at presentation. All patients had limitation of abduction and internal rotation. Eight patients (53.33%) had pain at the hip and 12 patients (80%) had limp. Mean time between diagnosis and corrective surgery was 3 weeks. RESULTS: The evaluation was done using caput index (CI) and epiphyseal quotient (EQ) and articulotrochanteric distance radiologically, range of motion and Harris Hip Score for clinical outcome. All the measurements were carried out on pre- and postoperative X-rays after 3 years followup and compared with the contralateral normal hip. After a mean followup period of 3.4 years, we noted the statistically significant difference between pre- and postoperative values. We noted that all (100%) children in Stage IB, IIA and 50% children in Stage IIB achieved satisfactory results. There was a significant change (P = 0.000) in CI among all the patients after surgery. The final EQ after 3 years of VDRO was 0.606 and was significant (P = 0.0000). CONCLUSION: In our opinion, based on the encouraging short term radiological and clinical outcomes, VDRO may be regarded as a treatment procedure for late presenting Perthes disease in stage IB, IIA, IIB.

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In humans and other animals, behavioural variation in learning has been associated with variation in neural features like morphology and myelination. By contrast, it is essentially unknown whether cognitive performance scales with electrophysiological properties of individual neurons. Birdsong learning offers a rich system to investigate this topic as song acquisition is similar to human language learning. Here, we address the interface between behavioural learning and neurophysiology in a cohort of wild-caught, hand-reared songbirds (swamp sparrows, Melospiza georgiana). We report the discovery in the forebrain HVC of sensorimotor 'bridge' neurons that simultaneously and selectively represent two critical learning-related schemas: the bird's own song, and the specific tutor model from which that song was copied. Furthermore, the prevalence and response properties of bridge neurons correlate with learning ability - males that copied tutor songs more accurately had more bridge neurons. Our results are consistent with the hypothesis that accurate imitative learning depends on a successful bridge, within single cortical neurons, between the representation of learning models and their sensorimotor copies. Whether such bridge neurons are a necessary mechanism for accurate learning or an outcome of learning accuracy is unknown at this stage, but can now be addressed in future developmental studies.

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PURPOSE: Pro-inflammatory cytokines such as Interleukin-17A (IL17A) and Interleukin-32 (IL32), known to enhance natural killer and T cell responses, are also elevated in human malignancies and linked to poor clinical outcomes. To address this paradox, we evaluated relation between IL17A and IL32 expression and other inflammation- and T cell response-associated genes in breast tumors. METHODS: TaqMan-based gene expression analysis was carried out in seventy-eight breast tumors. The association between IL17A and IL32 transcript levels and T cell response genes, ER status as well as lymph node status was also examined in breast tumors from TCGA dataset. RESULTS: IL17A expression was detected in 32.7% ER-positive and 84.6% ER-negative tumors, with higher expression in the latter group (26.2 vs 7.1-fold, p < 0.01). ER-negative tumors also showed higher expression of IL32 as opposed to ER-positive tumors (8.7 vs 2.5-fold, p < 0.01). Expression of both IL17A and IL32 genes positively correlated with CCL5, GNLY, TBX21, IL21 and IL23 transcript levels (p < 0.01). Amongst ER-positive tumors, higher IL32 expression significantly correlated with lymph node metastases (p < 0.05). Conversely, in ER-negative subtype, high IL17A and IL32 expression was seen in patients with negative lymph node status (p < 0.05). Tumors with high IL32 and IL17A expression showed higher expression of TH1 response genes studied, an observation validated by similar analysis in the TCGA breast tumors (n=1041). Of note, these tumors were characterized by low expression of a potentially immunosuppressive isoform of IL32 (IL32γ). CONCLUSION: These results suggest that high expression of both IL17A and IL32 leads to enhancement of T cell responses. Our study, thus, provides basis for the emergence of strong T cell responses in an inflammatory milieu that have been shown to be associated with better prognosis in ER-negative breast cancer.

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PURPOSE: Tibial fracture is the most common long bone fracture. Distal third tibial fractures are challenging though open reduction and plating can result in anatomical reduction and rigid fixation. This paper aimed to evaluate and compare the results of medial and lateral locking compression plates for distal third tibial fractures. METHODS: This prospective clinical study involved 36 patients with distal tibial fractures admitted in Department of Orthopaedics, Sawai Mansingh Medical College & Affiliated Hospital, Jaipur, India, from June 2011 to May 2012, including 29 closed fractures and 7 open fractures at the mean age of 38.9 years. Thirty-six patients were divided equally into two groups based on treatment method, including medial plating group (18 patients) and lateral plating group (18 patients). They were followed up for at least 5 months after discharge. The functional outcomes were evaluated using Tenny and Wiss clinical assessment criteria. RESULTS: Malunion was found in 3 cases of medial plating group and in 1 case of lateral plating group. In the medial plating group, there were 5 cases of superficial infections, 1 deep infection, 1 nonunion and 3 wound dehiscence. In the lateral plating group, there was 1 case of superficial infections, 1 deep infection and 1 nonunion. In the lateral plating group, 4 patients reported feeling the plates and screws but none of them asked to remove the hardware. In the medial plating group, 9 patients reported symptomatic hardware problems and 7 asked to remove the hardware. The number of cases graded as excellent/good/fair was 1/8/7 in the medial plating group and 3/7/7 in the lateral plating group respectively. In the medial plating group, the final range of motion was 17.2° in ankle dorsiflexion and 30.7° in ankle plantar flexion. In the lateral plating group, the final range of motion was 19° in ankle dorsiflexion and 34.2° in ankle plantar flexion. CONCLUSION: Lateral plating of distal tibia is safe and feasible, which can provide biological fixation and prevent the soft tissue complications associated with medial plating.

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Open fractures with bone loss are a common occurrence following high energy trauma. But usually the bone fragments are lost on the roadside and are not usable. We report a patient who was involved in a head-on collision between two wheelers and presented with a bone fragment embedded in his thigh. Radiological survey revealed no bony injury in that patient. Another patient, who presented at the same time, sustained a segmental fracture of shaft femur and was found to have lost a bone fragment that was similar to the one found in previous patient. CT scan with 3D reconstruction revealed this missing fragment to be the same as that found in previous patient. Both patients had a history of head-on collision while travelling on a two-wheeler. Present case report throws some highlights on the probable mechanism of injury.

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