Establishment and validation of a 2D primary gill cell culture of the sevenband grouper (Hyporthodus septemfasciatus).

A 2D primary gill cell culture system of the sevenband grouper (Hyporthodus septemfasciatus) was established to validate the pathogenesis of nervous necrosis virus (NNV) as observed in previous studies. This system, developed using the double-seeded insert (DSI) technique, yielded confluent cell layers. Upon challenge with NNV in a setup containing both autoclaved salt water and L15 media in the apical compartment, viral replication akin to that anticipated based on previous studies was observed. Consequently, we advocate for the utilization of primary gill cell culture as a viable alternative to conventional methodologies for investigating host pathogen interactions.

Digital analysis of the prostate tumor microenvironment with high-order chromogenic multiplexing.

As our understanding of the tumor microenvironment grows, the pathology field is increasingly utilizing multianalyte diagnostic assays to understand important characteristics of tumor growth. In clinical settings, brightfield chromogenic assays represent the gold-standard and have developed significant trust as the first-line diagnostic method. However, conventional brightfield tests have been limited to low-order assays that are visually interrogated. We have developed a hybrid method of brightfield chromogenic multiplexing that overcomes these limitations and enables high-order multiplex assays. However, how compatible high-order brightfield multiplexed images are with advanced analytical algorithms has not been extensively evaluated. In the present study, we address this gap by developing a novel 6-marker prostate cancer assay that targets diverse aspects of the tumor microenvironment such as prostate-specific biomarkers (PSMA and p504s), immune biomarkers (CD8 and PD-L1), a prognostic biomarker (Ki-67), as well as an adjunctive diagnostic biomarker (basal cell cocktail) and apply the assay to 143 differentially graded adenocarcinoma prostate tissues. The tissues were then imaged on our spectroscopic multiplexing imaging platform and mined for proteomic and spatial features that were correlated with cancer presence and disease grade. Extracted features were used to train a UMAP model that differentiated healthy from cancerous tissue with an accuracy of 89% and identified clusters of cells based on cancer grade. For spatial analysis, cell-to-cell distances were calculated for all biomarkers and differences between healthy and adenocarcinoma tissues were studied. We report that p504s positive cells were at least 2× closer to cells expressing PD-L1, CD8, Ki-67, and basal cell in adenocarcinoma tissues relative to the healthy control tissues. These findings offer a powerful insight to understand the fingerprint of the prostate tumor microenvironment and indicate that high-order chromogenic multiplexing is compatible with digital analysis. Thus, the presented chromogenic multiplexing system combines the clinical applicability of brightfield assays with the emerging diagnostic power of high-order multiplexing in a digital pathology friendly format that is well-suited for translational studies to better understand mechanisms of tumor development and growth.

Malignant Peripheral Nerve Sheath Tumor of the Hand: A Case Report.

An uncommon and aggressive type of soft tissue sarcoma that develops from peripheral nerves is known as a malignant peripheral nerve sheath tumor (MPNST). It is typically associated with neurofibromatosis type 1. Hence, this case report presents a case of a 42-year-old woman with a mass over the palmar aspect of her right hand that had been slowly growing over the previous year. Complete resection of the tumor with groin flap was performed followed by adjuvant radiotherapy. Over the last year, the patient has been monitored on an outpatient basis without displaying any signs of a local recurrence.

Spontaneous Intestinal Perforation in Neonates Involving the Cecum: A Case Report.

Neonatal spontaneous colonic perforation in term neonates is a rare phenomenon, with caecal perforation being seldom reported. Therefore, this case report presents a rare case of spontaneous caecal perforation in a term neonate who presented with vomiting and abdominal distension on day two of life. On exploration, a single large full­thickness cecum perforation was noted. Histopathologic samples were negative for necrotizing enterocolitis and Hirschsprung's disease. Clinical awareness of this rare entity could help in preventing delays in imaging and prompt surgical management.

Monitoring Cardiovascular Problems in Heart Patients Using Machine Learning.

The World Health Organization reports that heart disease is the most common cause of death globally, accounting for 17.9 million fatalities annually. The fundamentals of a cure, it is thought, are important symptoms and recognition of the illness. Traditional techniques are facing many challenges, ranging from delayed or unnecessary treatment to incorrect diagnoses, which can affect treatment progress, increase the bill, and give the disease more time to spread and harm the patient's body. Such errors could be avoided and minimized by employing ML and AI techniques. Many significant efforts have been made in recent years to increase computer-aided diagnosis and detection applications, which is a rapidly growing area of research. Machine learning algorithms are especially important in CAD, which is used to detect patterns in medical data sources and make nontrivial predictions to assist doctors and clinicians in making timely decisions. This study aims to develop multiple methods for machine learning using the UCI set of data based on individuals' medical attributes to aid in the early detection of cardiovascular disease. Various machine learning techniques are used to evaluate and review the results of the UCI machine learning heart disease dataset. The proposed algorithms had the highest accuracy, with the random forest classifier achieving 96.72% and the extreme gradient boost achieving 95.08%. This will assist the doctor in taking appropriate actions. The proposed technology will only be able to determine whether or not a person has a heart issue. The severity of heart disease cannot be determined using this method.

Long-term changes in thickness, live/dead bacterial ratio, and mineral content in biofilm on ceramic and stainless steel orthodontic attachments.

PURPOSE: Fixed orthodontic appliances induce biofilm deposition, which harbors a microbial population harmful to the periodontal health of the individual. The present study evaluated the changes in thickness, live/dead bacterial ratio, and mineral content in dental biofilm over 6 months in patients with either stainless steel or ceramic orthodontic attachments. METHODS: Eighty patients who require fixed orthodontic appliance treatment with first premolar extraction for correcting their malocclusion were selected and bonded with either stainless steel or ceramic orthodontic attachments on the buccal side. The attached buttons were retrieved at different periods-1 week, 1 month, 3 months, and 6 months. They were stained and visualized through confocal microscopy to detect biofilm thickness and the ratio of live/dead bacteria. X­ray diffraction was used to identify the presence of calcium and phosphorous. RESULTS: Ceramic attachments showed a greater increase in biofilm thickness in comparison to stainless steel attachments except in the initial 1­week evaluation. A higher live/dead bacterial ratio was observed in stainless steel attachments than in their ceramic counterparts at all four evaluation periods. Both stainless steel and ceramic surfaces exhibited the presence of mineral deposition (calcium and phosphorous) at all periods. CONCLUSIONS: More biofilm adhesion was observed over ceramic surfaces than over stainless steel orthodontic attachments. Stainless steel attachments exhibited biofilm with a higher live/dead bacterial ratio than their ceramic counterparts at all evaluation periods. The presence of calcium and phosphorous in the adhered biofilm, pointing toward its calcification process, was identified.

A Giant Epidermal Cyst in the Gluteal Region: A Case Report.

Epidermal cysts are the most commonly occurring cysts in the subcutaneous plane and are usually asymptomatic, small, and slow growing. When the size of an epidermal cyst is greater than 5 cm, it is considered a giant epidermal cyst. Common etiologies include sun-damaged skin and acne vulgaris, and they can appear anywhere in the body, but preferably over the face, neck, and trunk. Unusual sites include the breast, penis, spleen, bones, subungual region, palms, soles, and buttocks. In this report, we presented the case of a 31-year-old female having a large, painless, gradually progressing swelling for two years in the left gluteal region, which was insidious in onset and slow-growing. The patient eventually described a discomfort that made it impossible for her to sit for long hours or sleep in a supine position. Clinical examination revealed circumscribed mass over the left gluteal region from which diagnosis of giant lipoma was confirmed, but due to its large size and involvement of the whole left buttock, we considered it important to perform an ultrasound examination to reinforce our diagnosis, which revealed a large cystic mass in the left gluteal subcutaneous plane that was excised. As a definitive management, surgery was performed with excision of the swelling, which was extracted in toto and was identified as a cyst, and on histopathological examination, the cyst wall was lined by stratified squamous epithelium. Hence, this case report highlights a rare case of a giant epidermal cyst in the gluteal region.

Temperature dependent cellular, and epigenetic regulatory mechanisms underlying the antiviral immunity in sevenband grouper to nervous necrosis virus infection.

Changes in the thermal optima of fish impacts changes in the physiology and immune response associated with infections. The present study showed that at suboptimal temperatures (17 °C), the host tries to evade viral infection by downregulating the inflammatory response through enhanced neuronal protection. There was significantly less abundance of IgM + B cells in the 17 °C group compared to that in the 25 °C group. An increased macrophage population (Iba1+) during the survival phase in fish challenged at 25 °C demonstrated inflammation. Optimal temperature challenge activated virus-induced senescence in brain cells, demonstrated with a heterochromatin-associated H3K9me3 histone mark. There was an abundant expression of anti-inflammatory cytokines in the brain of fish at the suboptimal challenge. Besides the cytokines, the expression of BDNF was significantly higher in the suboptimally challenged group, suggesting that its neuronal protection activity following NNV infection is mediated through TGFß. The suboptimal challenge resulted in H3k9ac displaying transcriptional competency, activation of trained immunity H3K4me3, and enrichment of H3 histone-lysine-4 monomethylation (H3K4me1), resulting in a robust re-stimulatory immune response. The observations from the H4 modifications showed that besides H4K12ac and H4K20m3, all the assayed modifications were significantly higher in suboptimal convalescent fishes. The suboptimally challenged fish acquired more methylation along cytosine residues than the optimally infected fish. Together, these observations suggest that optimal temperature results in an immune priming effect, whereas the protection enabled in suboptimal convalescent fishes is operated through epigenetically controlled trained immune functions.

Alginate as a Promising Biopolymer in Drug Delivery and Wound Healing: A Review of the State-of-the-Art.

Biopolymeric nanoparticulate systems hold favorable carrier properties for active delivery. The enhancement in the research interest in alginate formulations in biomedical and pharmaceutical research, owing to its biodegradable, biocompatible, and bioadhesive characteristics, reiterates its future use as an efficient drug delivery matrix. Alginates, obtained from natural sources, are the colloidal polysaccharide group, which are water-soluble, non-toxic, and non-irritant. These are linear copolymeric blocks of α-(1→4)-linked l-guluronic acid (G) and ß-(1→4)-linked d-mannuronic acid (M) residues. Owing to the monosaccharide sequencing and the enzymatically governed reactions, alginates are well-known as an essential bio-polymer group for multifarious biomedical implementations. Additionally, alginate's bio-adhesive property makes it significant in the pharmaceutical industry. Alginate has shown immense potential in wound healing and drug delivery applications to date because its gel-forming ability maintains the structural resemblance to the extracellular matrices in tissues and can be altered to perform numerous crucial functions. The initial section of this review will deliver a perception of the extraction source and alginate's remarkable properties. Furthermore, we have aspired to discuss the current literature on alginate utilization as a biopolymeric carrier for drug delivery through numerous administration routes. Finally, the latest investigations on alginate composite utilization in wound healing are addressed.

Poly(N-isopropylacrylamide)-Based Hydrogels for Biomedical Applications: A Review of the State-of-the-Art.

A prominent research topic in contemporary advanced functional materials science is the production of smart materials based on polymers that may independently adjust their physical and/or chemical characteristics when subjected to external stimuli. Smart hydrogels based on poly(N-isopropylacrylamide) (PNIPAM) demonstrate distinct thermoresponsive features close to a lower critical solution temperature (LCST) that enhance their capability in various biomedical applications such as drug delivery, tissue engineering, and wound dressings. Nevertheless, they have intrinsic shortcomings such as poor mechanical properties, limited loading capacity of actives, and poor biodegradability. Formulation of PNIPAM with diverse functional constituents to develop hydrogel composites is an efficient scheme to overcome these defects, which can significantly help for practicable application. This review reports on the latest developments in functional PNIPAM-based smart hydrogels for various biomedical applications. The first section describes the properties of PNIPAM-based hydrogels, followed by potential applications in diverse fields. Ultimately, this review summarizes the challenges and opportunities in this emerging area of research and development concerning this fascinating polymer-based system deep-rooted in chemistry and material science.

NLRC3 attenuates antiviral immunity and activates inflammasome responses in primary grouper brain cells following nervous necrosis virus infection.

NLRC3 is identified as a unique regulatory NLR involved in the modulation of cellular processes and inflammatory responses. In this study, a novel Nod like receptor C3 (NLRC3) was functionally characterized from seven band grouper in the context of nervous necrosis virus infection. The grouper NLRC3 is highly conserved and homologous with other vertebrate proteins with a NACHT domain and a C-terminal leucine-rich repeat (LRR) domain and an N-terminal CARD domain. Quantitative gene expression analysis revealed the highest mRNA levels of NLRC3 were in the brain and gill followed by the spleen and kidney following NNV infection. Overexpression of NLRC3 augmented the NNV replication kinetics in primary grouper brain cells. NLRC3 attenuated the interferon responses in the cells following NNV infection by impacting the TRAF6/NF-κB activity and exhibited reduced IFN sensitivity, ISRE promoter activity, and IFN pathway gene expression. In contrast, NLRC3 expression positively regulated the inflammasome response and pro-inflammatory gene expression during NNV infection. NLRC3 negatively regulates the PI3K-mTOR axis and activated the cellular autophagic response. Delineating the complexity of NLRC3 regulation of immune response in the primary grouper brain cells following NNV infection suggests that the protein acts as a virally manipulated host factor that negatively regulated the antiviral immune response to augment the NNV replication.

Tufts Dental Database: A Multimodal Panoramic X-Ray Dataset for Benchmarking Diagnostic Systems.

The application of Artificial Intelligence in dental healthcare has a very promising role due to the abundance of imagery and non-imagery-based clinical data. Expert analysis of dental radiographs can provide crucial information for clinical diagnosis and treatment. In recent years, Convolutional Neural Networks have achieved the highest accuracy in various benchmarks, including analyzing dental X-ray images to improve clinical care quality. The Tufts Dental Database, a new X-ray panoramic radiography image dataset, has been presented in this paper. This dataset consists of 1000 panoramic dental radiography images with expert labeling of abnormalities and teeth. The classification of radiography images was performed based on five different levels: anatomical location, peripheral characteristics, radiodensity, effects on the surrounding structure, and the abnormality category. This first-of-its-kind multimodal dataset also includes the radiologist's expertise captured in the form of eye-tracking and think-aloud protocol. The contributions of this work are 1) publicly available dataset that can help researchers to incorporate human expertise into AI and achieve more robust and accurate abnormality detection; 2) a benchmark performance analysis for various state-of-the-art systems for dental radiograph image enhancement and image segmentation using deep learning; 3) an in-depth review of various panoramic dental image datasets, along with segmentation and detection systems. The release of this dataset aims to propel the development of AI-powered automated abnormality detection and classification in dental panoramic radiographs, enhance tooth segmentation algorithms, and the ability to distill the radiologist's expertise into AI.

Nanoparticle-Based Drug Delivery System: The Magic Bullet for the Treatment of Chronic Pulmonary Diseases.

Chronic pulmonary diseases encompass different persistent and lethal diseases, including chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), cystic fibrosis (CF), asthma, and lung cancers that affect millions of people globally. Traditional pharmacotherapeutic treatment approaches (i.e., bronchodilators, corticosteroids, chemotherapeutics, peptide-based agents, etc.) are not satisfactory to cure or impede diseases. With the advent of nanotechnology, drug delivery to an intended site is still difficult, but the nanoparticle's physicochemical properties can accomplish targeted therapeutic delivery. Based on their surface, size, density, and physical-chemical properties, nanoparticles have demonstrated enhanced pharmacokinetics of actives, achieving the spotlight in the drug delivery research field. In this review, the authors have highlighted different nanoparticle-based therapeutic delivery approaches to treat chronic pulmonary diseases along with the preparation techniques. The authors have remarked the nanosuspension delivery via nebulization and dry powder carrier is further effective in the lung delivery system since the particles released from these systems are innumerable to composite nanoparticles. The authors have also outlined the inhaled particle's toxicity, patented nanoparticle-based pulmonary formulations, and commercial pulmonary drug delivery devices (PDD) in other sections. Recently advanced formulations employing nanoparticles as therapeutic carriers for the efficient treatment of chronic pulmonary diseases are also canvassed.

Morphological alterations, activity, mRNA fold changes, and aging changes before and after orthodontic force application in young and adult human-derived periodontal ligament cells.

BACKGROUND: The response of periodontal ligament cells (PDLC) from adult subjects in comparison to those obtained from younger ones to mechanical forces has been a matter of interest recently because of induced senescent changes. This study evaluated and compared cell surface changes and activity, integrin beta 1, and ß-actin mRNA fold changes as well as klotho protein secretion capabilities of PDLC from young and adult donors before and after subjecting to orthodontic forces. METHODS: A total of 40 subjects with bimaxillary dentoalveolar protrusion requiring extraction of first premolars for orthodontic treatment were selected and divided into two groups. Force ranging from 80 to 90 g was applied to maxillary first premolars and extraction was carried out at two different time periods-pre-treatment (control group) and 28 days after force application (experimental group). Periodontal ligament was obtained, and cell surface changes and activity were observed with atomic force microscopy (AFM) and fluorescent tagging. mRNA fold change of integrin beta-1 and ß-actin mRNA, as well as beta-galactosidase assay, was performed, and levels of klotho protein were evaluated. RESULTS: AFM nanoindentation and fluorescent tagging indicated increased surface morphological changes in younger cells compared to adult ones. We observed a decrease in integrin beta 1 but an increase in ß-actin mRNA levels in PDLC obtained from younger subjects compared to adults, while an increase was observed in SA-ß-GAL from adult cells. The level of klotho protein was lower in adult cells in comparison to younger ones. LIMITATIONS: Large sample studies are required to find out a variation in aging characteristics between young and adult PDLC. CONCLUSIONS: The study observed significant differences between PDLC obtained from younger and adult subjects in response to orthodontic force application.

Effect of Non-Newtonian Dynamics on the Clearance of Mucus From Bifurcating Lung Airway Models.

Mucus hypersecretion is a common pathophysiological manifestation of several obstructive airway diseases in which the mucociliary clearance is impaired, and the airflow generated by a cough or a forced expiratory maneuver called the huff is primarily responsible for clearing mucus. This airflow driven clearance of mucus is a complex process that is affected by the mucus rheology, airflow rate, airway geometry, and gravity. This study examines the role of mucus rheology in the transport and distribution of mucus in idealized 3D airway geometries. The complex air-mucus interface was tracked by the volume-of-fluid (VOF) model, and the turbulence in the core airflow was modeled using the k-ω shear stress transport (SST) model. Mucus was modeled as a shear-thinning liquid by using a power-law model. The computational model was validated using in vitro experimental data available in the literature. Gravity-dominated eccentric core-annular flow was observed with the core biased toward the outer wall in the inclined daughter branches of the bifurcation models, which transitions into concentric core-annular flow in the trachea. The increase in tangential shear at the interface due to the secondary flow structures developed in the flow divider location resulted in a region of enhanced mucus clearance with reduced mucus layer thickness. Secondary flow developed due to the curvature in the airway geometry resulted in a local redistribution of mucus that reduced the eccentricity. The accumulation of mucus around the carinal ridges and the regions with reduced clearance are sites with the potential for microbial growth.

A Rare Case of Parvovirus B19 Infection Induced Paroxysmal Cold Hemoglobinuria in an Adult Female.

Paroxysmal cold hemoglobinuria (PCH) is a rare form of autoimmune hemolytic anemia (AIHA). PCH occurs in acute and chronic forms. The main risk factors for PCH include viral infections, vaccination, and syphilis. PCH presentations are common in the pediatric population. The occurrence of PCH following parvovirus B19 infection in adults is rare. We report a case of a 23-year-old female who presented with giddiness, fatigue, greying of vision, and presyncope for four days, on subsequent evaluation was found to have evidence of hemolysis and bone marrow suppression. Parvoviral intranuclear inclusions were detected in bone marrow biopsy and parvoviral B19 IgM antibody was detected. Donath Landsteiner antibody test was also positive. Hence a diagnosis of PCH secondary to parvovirus B19 infection was made. She was started on pulse dose steroids and intravenous immunoglobulin (IVIG) and showed significant improvement.

An Unusual Presentation of Adult-Onset Still's Disease as Hemophagocytic Lymphohistiocytosis in a Male Patient.

Hemophagocytic lymphohistiocytosis (HLH) is an aggressive and potentially fatal condition characterized by immune activation and multi-organ dysfunction. HLH can be inherited in an autosomal recessive fashion, but can also be secondary to infections, malignancy, immunosuppression, and autoimmune conditions. Adult-onset Still's disease (AOSD) is an autoimmune disorder that can predispose patients to HLH. AOSD, similar to other autoimmune conditions, is more common in females than males. However, the occurrence of AOSD in males and subsequent predisposition to HLH is rarely reported. We report the case of a 23-year-old male patient who presented with fever, joint pain, and rash for 20 days. On evaluation, he fulfilled the diagnostic criteria for AOSD and HLH, and a diagnosis of HLH secondary to AOSD was made. He was treated with pulse dose steroids and gradually tapered. AOSD and HLH have overlapping clinical and laboratory features and hence their co-occurrence poses diagnostic challenges. The mortality rate of HLH is high and hence prompt initiation of treatment is of utmost importance.

A Comprehensive Database for Benchmarking Imaging Systems.

Cross-modality face recognition is an emerging topic due to the wide-spread usage of different sensors in day-to-day life applications. The development of face recognition systems relies greatly on existing databases for evaluation and obtaining training examples for data-hungry machine learning algorithms. However, currently, there is no publicly available face database that includes more than two modalities for the same subject. In this work, we introduce the Tufts Face Database that includes images acquired in various modalities: photograph images, thermal images, near infrared images, a recorded video, a computerized facial sketch, and 3D images of each volunteer's face. An Institutional Research Board protocol was obtained and images were collected from students, staff, faculty, and their family members at Tufts University. The database includes over 10,000 images from 113 individuals from more than 15 different countries, various gender identities, ages, and ethnic backgrounds. The contributions of this work are: 1) Detailed description of the content and acquisition procedure for images in the Tufts Face Database; 2) The Tufts Face Database is publicly available to researchers worldwide, which will allow assessment and creation of more robust, consistent, and adaptable recognition algorithms; 3) A comprehensive, up-to-date review on face recognition systems and face datasets.

Initial Evaluation of Computer-Assisted Radiologic Assessment for Renal Mass Edge Detection as an Indication of Tumor Roughness to Predict Renal Cancer Subtypes.

OBJECTIVE: To develop software to assess the potential aggressiveness of an incidentally detected renal mass using images. METHODS: Thirty randomly selected patients who underwent nephrectomy for renal cell carcinoma (RCC) had their images independently reviewed by engineers. Tumor "Roughness" was based on image algorithm of tumor topographic features visualized on computed tomography (CT) scans. Univariant and multivariant statistical analyses are utilized for analysis. RESULTS: We investigated 30 subjects that underwent partial or radical nephrectomy. After excluding poor image-rendered images, 27 patients remained (benign cyst = 1, oncocytoma = 2, clear cell RCC = 15, papillary RCC = 7, and chromophobe RCC = 2). The mean roughness score for each mass is 1.18, 1.16, 1.27, 1.52, and 1.56 units, respectively (p < 0.004). Renal masses were correlated with tumor roughness (Pearson's, p=0.02). However, tumor size itself was larger in benign tumors (p=0.1). Linear regression analysis noted that the roughness score is the most influential on the model with all other demographics being equal including tumor size (p=0.003). CONCLUSION: Using basic CT imaging software, tumor topography ("roughness") can be quantified and correlated with histologies such as RCC subtype and could lead to determining aggressiveness of small renal masses.