Histology Imaging Research Articles

VEGFC Overexpression in Kidney Progenitor Cells Is a Model of Renal Lymphangiectasia.

Lymphangiogenesis is believed to be a protective response in the setting of multiple forms of kidney injury and mitigates the progression of interstitial fibrosis. To augment this protective response, promoting kidney lymphangiogenesis is being investigated as a potential treatment to slow the progression of kidney disease. As injury-related lymphangiogenesis is driven by signaling from the receptor VEGFR3 (vascular endothelial growth factor receptor 3) in response to the cognate growth factor VEGF (vascular endothelial growth factor)-C released by tubular epithelial cells, this signaling pathway is a candidate for future kidney therapeutics. However, the consequences to kidney development and function to targeting this signaling pathway remain poorly defined. We generated a new mouse model expressing Vegfc under regulation of the nephron progenitor Six2Cre driver strain (Six2Vegfc). Mice underwent a detailed phenotypic evaluation. Whole kidneys were processed for histology and 3-dimensional imaging. Six2Vegfc mice had reduced body weight and kidney function compared with littermate controls. Six2Vegfc kidneys demonstrated large peripelvic fluid-filled lesions with distortion of the pelvicalcyceal system which progressed in severity with age. Three-dimensional imaging showed a 3-fold increase in total cortical vascular density. Histology confirmed a substantial increase in LYVE1+ (lymphatic vessel endothelial hyaluronan receptor-1)/PDPN+ (podoplanin)/VEGFR3+ lymphatic capillaries extending alongside EMCN+ (endomucin) peritubular capillaries. There was no change in EMCN+ peritubular capillary density. Kidney lymphatic density was robustly increased in the Six2Vegfc mice. There were no changes in peritubular blood capillary density despite these endothelial cells also expressing VEGFR3. The model resulted in malformation of the lymphatic hilar plexus, resulting in severe hydronephrosis that resembled a human condition termed renal lymphangiectasia. This study defines the vascular consequences of augmenting VEGFC signaling during kidney development and provides new insight into human renal lymphatic malformations.

Large depth-of-field ultraviolet-visible photoacoustic histologic and microvascular imaging in vivo

Photoacoustic microscopy (PAM) can image diverse biological microstructures by harnessing characteristic optical absorption spectra of intrinsic nonfluorescent biomolecules. We incorporate ultraviolet (UV) and visible pulsed lasers into a PAM to develop an UV-visible PAM for label-free histologic and microvascular imaging, where the cell nuclei and blood vessels are specifically captured at high contrast relying on strong optical absorption of DNA/RNA and hemoglobin at 266 and 532 nm wavelengths, respectively. Moreover, two diffractive optical elements operating at UV and visible spectra are designed for engineering the excitation beams, significantly enlarging depths of field (DOFs > 200 μm). The UV-visible PAM demonstrates combined capabilities of dual imaging contrast, elongated DOFs, and micrometer-scale lateral resolution for delineating the spatial microarchitectures of both cell nuclei and blood vessels that are at different depth locations in the biological specimens with uneven surfaces. Longitudinal monitoring of the trauma is performed in mouse ear in vivo. Potentially, our UV-visible PAM could offer comprehensive histologic and microvascular information in a broad range of biomedical studies.

Intraoperative Label-Free Tissue Diagnostics using a Stimulated Raman Histology Imaging System with Artificial Intelligence: An Initial Experience

Intraoperative Label-Free Tissue Diagnostics using a Stimulated Raman Histology Imaging System with Artificial Intelligence: An Initial Experience

Seq-Scope: repurposing Illumina sequencing flow cells for high-resolution spatial transcriptomics.

Spatial transcriptomics technologies aim to advance gene expression studies by profiling the entire transcriptome with intact spatial information from a single histological slide. However, the application of spatial transcriptomics is limited by low resolution, limited transcript coverage, complex procedures, poor scalability and high costs of initial setup and/or individual experiments. Seq-Scope repurposes the Illumina sequencing platform for high-resolution, high-content spatial transcriptome analysis, overcoming these limitations. It offers submicrometer resolution, high capture efficiency, rapid turnaround time and precise annotation of histopathology at a much lower cost than commercial alternatives. This protocol details the implementation of Seq-Scope with an Illumina NovaSeq 6000 sequencing flow cell, allowing the profiling of multiple tissue sections in an area of 7 mm × 7 mm or larger. We describe the preparation of a fresh-frozen tissue section for both histological imaging and sequencing library preparation and provide a streamlined computational pipeline with comprehensive instructions to integrate histological and transcriptomic data for high-resolution spatial analysis. This includes the use of conventional software tools for single-cell and spatial analysis, as well as our recently developed segmentation-free method for analyzing spatial data at submicrometer resolution. Aside from array production and sequencing, which can be done in batches, tissue processing, library preparation and running the computational pipeline can be completed within 3 days by researchers with experience in molecular biology, histology and basic Unix skills. Given its adaptability across various biological tissues, Seq-Scope establishes itself as an invaluable tool for researchers in molecular biology and histology.

Applying Deep-Learning Algorithm Interpreting Kidney, Ureter, and Bladder (KUB) X-Rays to Detect Colon Cancer.

Early screening is crucial in reducing the mortality of colorectal cancer (CRC). Current screening methods, including fecal occult blood tests (FOBT) and colonoscopy, are primarily limited by low patient compliance and the invasive nature of the procedures. Several advanced imaging techniques such as computed tomography (CT) and histological imaging have been integrated with artificial intelligence (AI) to enhance the detection of CRC. There are still limitations because of the challenges associated with image acquisition and the cost. Kidney, ureter, and bladder (KUB) radiograph which is inexpensive and widely used for abdominal assessments in emergency settings and shows potential for detecting CRC when enhanced using advanced techniques. This study aimed to develop a deep learning model (DLM) to detect CRC using KUB radiographs. This retrospective study was conducted using data from the Tri-Service General Hospital (TSGH) between January 2011 and December 2020, including patients with at least one KUB radiograph. Patients were divided into development (n = 28,055), tuning (n = 11,234), and internal validation (n = 16,875) sets. An additional 15,876 patients were collected from a community hospital as the external validation set. A 121-layer DenseNet convolutional network was trained to classify KUB images for CRC detection. The model performance was evaluated using receiver operating characteristic curves, with sensitivity, specificity, and area under the curve (AUC) as metrics. The AUC, sensitivity, and specificity of the DLM in the internal and external validation sets achieved 0.738, 61.3%, and 74.4%, as well as 0.656, 47.7%, and 72.9%, respectively. The model performed better for high-grade CRC, with AUCs of 0.744 and 0.674 in the internal and external sets, respectively. Stratified analysis showed superior performance in females aged 55-64 with high-grade cancers. AI-positive predictions were associated with a higher long-term risk of all-cause mortality in both validation cohorts. AI-enhanced KUB X-ray analysis can enhance CRC screening coverage and effectiveness, providing a cost-effective alternative to traditional methods. Further prospective studies are necessary to validate these findings and fully integrate this technology into clinical practice.

Rationale and study design of the international randomized control trial MYocarditis THerapy with Steroids: the MYTHS trial

Abstract Background Immunosuppression is generally suggested in patients with specific subtypes of acute myocarditis, such as eosinophilic myocarditis or giant cell myocarditis or associated with systemic autoimmune disorders. In contrast, no specific treatment for most acute myocarditis complicated by acute heart failure (HF) or cardiogenic shock is recommended beyond the management of acute HF and supportive therapy. Nevertheless, large registries commonly reported using corticosteroids in severe forms without data supporting this strategy. Purpose To assess the safety and efficacy of high-dose pulse intravenous (IV) corticosteroid therapy to treat patients with complicated/fulminant acute myocarditis based on clinical suspicion. Methods An international pragmatic investigator-initiated single-blinded randomized controlled trial that screens patients between 18 and 70 years admitted to the hospital for suspected acute myocarditis (cardiac symptoms onset within three weeks) complicated by acute HF/cardiogenic shock and left ventricular (LV) ejection fraction<41% without significant LV dilation is ongoing. Patients are randomized in a 1:1 ratio to pulsed IV methylprednisolone (1 g) in saline solution (250 mL) daily for three days or placebo (saline without methylprednisolone) on top of standard therapy and maximal supportive care. A final diagnosis of myocarditis is confirmed by histology or cardiac magnetic resonance imaging. A population of 288 patients has been planned to assess a reduction in the probability of reaching the primary endpoint from 25% in the placebo arm to 12% in the methylprednisolone arm with a power of 0.80 (Figure). The main analysis will be conducted on an intention-to-treat basis. Results The primary efficacy endpoint of the trial is defined as the time from randomization to the first event occurring within 6 months, including all-cause death, heart transplantation, or long-term LV assist device implant, or need for an upgrading of the temporary mechanical circulatory support (t-MCS), or ventricular tachycardia (VT)/fibrillation (VF) treated with direct current (DC) shock, or first rehospitalization due to HF or VT/VF, or advanced atrioventricular block. The safety endpoints include a composite of potential complications that can occur, including serious infections, acute gastrointestinal bleeding, and other complications that can be related to corticosteroid therapy or prolonged stay on t-MCS. There are 32 active centers, and 48 patients have been randomized since December 2021. The trial is expected to be completed by the end of 2028. Conclusion The results of the MYTHS trial can define the role of IV methylprednisolone in the initial treatment of patients with complicated forms of acute myocarditis that currently have an unacceptably high mortality rate.

Long-range enhancement for fluorescence and Raman spectroscopy using Ag nanoislands protected with column-structured silica overlayer

We demonstrate long-range enhancement of fluorescence and Raman scattering using a dense random array of Ag nanoislands (AgNIs) coated with column-structured silica (CSS) overlayer of over 100 nm thickness, namely, remote plasmonic-like enhancement (RPE). The CSS layer provides physical and chemical protection, reducing the impact between analyte molecules and metal nanostructures. RPE plates are fabricated with high productivity using sputtering and chemical immersion in gold(I)/halide solution. The RPE plate significantly enhances Raman scattering and fluorescence, even without proximity between analyte molecules and metal nanostructures. The maximum enhancement factors are 107-fold for Raman scattering and 102-fold for fluorescence. RPE is successfully applied to enhance fluorescence biosensing of intracellular signalling dynamics in HeLa cells and Raman histological imaging of oesophagus tissues. Our findings present an interesting deviation from the conventional near-field enhancement theory, as they cannot be readily explained within its framework. However, based on the phenomenological aspects we have demonstrated, the observed enhancement is likely associated with the remote resonant coupling between the localised surface plasmon of AgNIs and the molecular transition dipole of the analyte, facilitated through the CSS structure. Although further investigation is warranted to fully understand the underlying mechanisms, the RPE plate offers practical advantages, such as high productivity and biocompatibility, making it a valuable tool for biosensing and biomolecular analysis in chemistry, biology, and medicine. We anticipate that RPE will advance as a versatile analytical tool for enhanced biosensing using Raman and fluorescence analysis in various biological contexts.

Ultrastructural evaluation of adverse effects on dentine formation from systemic fluoride application in an experimental mouse model.

Fluoride is widely used in dentistry for its caries prevention. To reduce dental caries, the optimal fluoride concentration of public water supplies in the United States is 0.7 ppm. However, excessive systemic fluoride consumption can lead to dental/enamel fluorosis. Numerous studies have explored the effects of fluoride on enamel and enamel-forming cells. However, research on systemic fluoride's impact on dentine is limited, particularly the effect of fluoride on the structure of the dentine-pulp complex. Therefore, this study aimed to identify how excessive fluoride affects dentine microstructure using an experimental mouse model. C57BL6/J male mice (6-9 weeks old) were randomized into four groups (Fluoride at 0, 50, 100, or 125 ppm in drinking water) (n = 4/group). Mice were provided water adlibitum for 6 weeks along with fluoride-free food. Thereafter, mandibular incisors were analysed. Enamel phenotypes were evaluated using light microscopy and quantitative light-induced fluorescence (QLF) to measure fluorosis levels. Dentine morphology was evaluated using micro-CT, scanning electron microscopy (SEM), SEM-EDX (energy-dispersive X-ray), microhardness test and histological imaging. Data were analysed using one-way ANOVA with Dunnett's multiple comparisons as a post hoc test and the Kruskal-Wallis test with Dunn's multiple comparisons post hoc test (p < .05). Mice treated with fluoride at 50-125 ppm developed enamel hypoplasia in their erupting incisors and micro-CT imaging revealed that fluoride 125 ppm caused external resorption of the growing incisor. Dentine mineral density, dentine volume decreased compared with the 0 ppm control, while pulp volume increased compared with the 0 ppm control group. SEM showed wider predentine layer and abnormalities in calcified matrix vesicles derived from odontoblasts in fluoride 100 and 125 ppm groups. Vickers microhardness of dentine significantly decreased in the high-dose group. Fluoride-induced dentine hypoplasia in a dose-dependent manner. Histological evaluation showed excessive fluoride 125 ppm induced micro abscess formation and inflammatory cell infiltration. Fluoride induced dentine dysplasia with a dentine microstructure resembling hypophosphatasia. High doses of systemic fluoride can cause dentine dysplasia. Both three-dimensional and microstructural analyses showed the structural, chemical and mechanical changes in the dentine and the mineralized tissue components, along with external resorption and pulp inflammation.

Effect of a novel cannulation device for vascular access on AVF maturation in a goat carotid-jugular fistula model.

The Ark is a 3-D printed titanium device designed to be implanted around the draining vein of an arteriovenous fistula (AVF) to facilitate vascular access. The purpose of this study was to assess AVF maturation after Ark implantation in a large animal model. End-to-side AVFs were created between the carotid artery and jugular vein in nine pygmy goats that included three control (AVF only) and six experimental (AVF and Ark device) animals. For experimental animals, an Ark device was implanted approximately 10 cm downstream of the anastomosis at the time of AVF creation. Postoperative ultrasounds and cannulations of the jugular vein fistula were performed over 12 months. At the conclusion of the study, the AVF was ligated and Ark devices along with a segment of the arterialized vein and surrounding tissues were explanted for gross and histological assessment. The control and experimental Ark groups exhibited increased dilation and flow as well as diminished depth underscoring the parallel developments in vascular attributes and AVF maturation between the two groups. Gross pathology, histology, and micro-CT imaging revealed intact endothelium, mature tissue integration throughout the porous Ark device, and no underlying stenosis. No adverse events such as foreign body reaction, skin or vessel erosion were identified. The study showed maturation without stenosis of the fistula in all animals. This study confirmed that the Ark device functions as a scaffold around the access vein, allows fistula maturation, and can be consistently cannulated without infiltrations over a 12-month period in a large animal model.

7703 Adrenal Cortical Carcinoma With Vascular Involvement - Complex Case Requiring Life-Saving Surgery With Bovine Pericardium- IVC Reconstruction

Abstract Disclosure: V. mehta: None. M. Renzu: None. K.N. Pereira: None. S. Sampath: None. S. Sruthi: None. E.B. Ruby: None. Adrenocortical carcinomas (ACCs) are rare endocrine tumors that exhibit unusual biological characteristics. Roughly 66% of cases are functional and have excessive hormone release causing classic symptoms of hypercortisolism, including plethora, diabetes, muscle weakness/atrophy, virilization, and osteoporosis. Few present with vague signs due to mass effects, including but not limited to abdominal/flank pain, fullness, or early satiety; the rest are incidental findings. We present a case of a 55-year-old female with no medical history presented following weeks of progressive abdominal pain with abdominal wall erythema, leg swelling, and dyspnea along with hirsutism, leukocytosis, elevated BNP, hematuria concerning for abdominal wall cellulitis and new-onset CHF. Initial CT chest revealed incompletely visualized incidental adrenal mass. Further investigations with CT abdomen and lab work revealed a left adrenal mass suspicious for ACC, extending into the left renal vein and IVC, elevated urine metanephrines and normetanephrine, and elevated cortisol levels with low ACTH. The dexamethasone suppression test confirmed inadequate cortisol suppression. Patient underwent aggressive surgery with exploratory laparotomy revealing tumor thrombus in left renal vein extending into Inferior vena cava (IVC). A left adrenalectomy, nephrectomy, and IVC reconstruction using a Bovine Pericardial Patch (BPP), which is majorly used for arterial reconstruction, was performed. Pathology results indicated ACC, measuring 13.5 cm, with extension into renal vein and vascular invasion, staged as pT4pN0 cM0 without evidence of deficient mismatch repair protein expression. ACCs are rare and profoundly aggressive malignancies with an incidence rate of 0.7–2.0 per million per year with bimodal age distribution. Disease peaks before the age of five and in the fourth to fifth decade of life, with increased incidence in females. Diagnosis cannot be conclusively established by a single histological technique, imaging, hormonal work-up, or immunohistochemical labeling, but surgical excision is the gold standard. Complete surgical resection is the only possible approach for curing or improving survival chances in patients with adrenal mass/metastasis. A retrospective case study of 36 venous reconstructions using BPP showed few complications like thrombosis in 14% (versus 11% in autologous venous repair), 11% reoperated, 3-4% developed partial thrombus and hematoma. Our case exemplifies the successful management of challenging ACC with vascular involvement using BPP, which is mostly used for arterial repairs and is now expanding to venous repairs. Outcomes stress the ongoing need for research and collaborative efforts to refine and standardize the ideal approach to ACC patients with IVC thrombosis and the use of BPP in venous repair. Presentation: 6/1/2024

5-Azacytidine treatment inhibits the development of lung cancer models via epigenetic reprogramming and activation of cellular pathways with anti-tumor activity.

Non-small cell lung cancer (NSCLC) treatment is challenged by late detection and limited therapeutic options. Aberrant DNA methylation, a common epigenetic alteration in NSCLC, offers new therapeutic avenues. This study aims to evaluate the combined effects of 5-Azacytidine (5-Aza), an epigenetic modifier, and ionizing radiation (IR) on NSCLC, exploring the underlying molecular mechanisms and therapeutic potential. In this study, we examined the effects of 5-Aza combined with IR in both in vitro and in vivo models of NSCLC. Five human NSCLC cell lines were treated with 5-Aza and IR. Cell viability, colony formation, wound healing, and transwell migration assays were performed to assess treatment effects. Microarray and qPCR analyses were conducted to identify gene expression changes. Additionally, subcutaneous and orthotopic xenograft models were used to evaluate the treatment's efficacy in vivo. Treatment with 5-Aza and IR resulted in significant reductions in cell viability, colony formation, and migration in NSCLC cell lines. Microarray analysis revealed significant changes in gene expression, including the upregulation of apoptosis-related genes and the downregulation of cell proliferation-related genes. In vivo studies demonstrated a notable reduction in tumor growth and metastasis in both subcutaneous and orthotopic NSCLC models following 5-Aza and IR treatment. Histological and bioluminescent imaging confirmed the therapeutic effects of the combined treatment. The combination of 5-Aza and IR shows promise as an effective treatment for NSCLC, enhancing apoptosis and reducing tumor growth through epigenetic modulation.

Repeated intrathecal injections of peripheral nerve-derived stem cell spheroids improve outcomes in a rat model of traumatic brain injury

BackgroundTraumatic brain injury (TBI) is a major cause of disability and mortality worldwide. However, existing treatments still face numerous clinical challenges. Building on our prior research showing peripheral nerve-derived stem cell (PNSC) spheroids with Schwann cell-like phenotypes can secrete neurotrophic factors to aid in neural tissue regeneration, we hypothesized that repeated intrathecal injections of PNSC spheroids would improve the delivery of neurotrophic factors, thereby facilitating the restoration of neurological function and brain tissue repair post-TBI.MethodsWe generated PNSC spheroids from human peripheral nerve tissue using suspension culture techniques. These spheroids were characterized using flow cytometry, immunofluorescence, and reverse-transcription polymerase chain reaction. The conditioned media were evaluated in SH-SY5Y and RAW264.7 cell lines to assess their effects on neurogenesis and inflammation. To simulate TBI, we established a controlled cortical impact (CCI) model in rats. The animals were administered intrathecal injections of PNSC spheroids on three occasions, with each injection spaced at a 3-day interval. Recovery of sensory and motor function was assessed using the modified neurological severity score (mNSS) and rotarod tests, while histological (hematoxylin and eosin, Luxol fast blue staining) and T2-weighted magnetic resonance imaging analyses, alongside immunofluorescence, were conducted to evaluate the recovery of neural structures and pathophysiology.ResultsPNSC spheroids expressed high levels of Schwann cell markers and neurotrophic factors, such as neurotrophin-3 and Ephrin B3. Their conditioned medium was found to promote neurite outgrowth, reduce reactive oxygen species-mediated cell death and inflammation, and influence M1-M2 macrophage polarization. In the CCI rat model, rats receiving repeated triple intrathecal injections of PNSC spheroids showed significant improvements in sensory and motor function, with considerable neural tissue recovery in damaged areas. Notably, this treatment promoted nerve regeneration, axon regrowth, and remyelination. It also reduced glial scar formation and inflammation, while encouraging angiogenesis.ConclusionOur findings suggest that repeated intrathecal injections of PNSC spheroids can significantly enhance neural recovery after TBI. This effect is mediated by the diverse neurotrophic factors secreted by PNSC spheroids. Thus, the strategy of combining therapeutic cell delivery with multiple intrathecal injections holds promise as a novel clinical treatment for TBI recovery.

Multiscale and multimodal evaluation of autosomal dominant polycystic kidney disease development

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most prevalent kidney genetic disorder, producing structural abnormalities and impaired function. This research investigates its evolution on mouse models, utilizing a combination of histology imaging, Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) to evaluate its progression thoroughly. ADPKD has been induced in mice via PKD2 gene knockout, followed by image acquisition at different stages. Histology data provides two-dimensional details, like the cystic area ratio, whereas CT and MRI facilitate three-dimensional temporal monitoring. Our approach allows to quantify the affected tissue at different disease stages through multiple quantitative metrics. A pivotal point is shown at approximately ten weeks after induction, marked by a swift acceleration in disease advancement, and leading to a notable increase in cyst formation. This multimodal strategy augments our comprehension of ADPKD dynamics and suggests the possibility of employing higher-resolution imaging in the future for more accurate volumetric analyses.

A novel m-xylylene-diamine/glucose based-supramolecular eutectogels with tissue clearing for three dimensional histological imaging

Hydrogel-based tissue clearing technologies have shown significant promise for deep-tissue imaging and subcellular-level optical 3D reconstruction of whole organs. This study proposes a novel approach utilizing a deep eutectic solvent (DES) formulated with glucose and m-xylylene-diamine (MXDA) to create a highly efficient tissue-clearing hydrogel system named the passive hydrogel clearing system (PHCS). PHCS achieved efficient tissue clearing through a single-step tissue gelation process. The resulting hydrogel-tissue complex exhibited thermoreversible properties, transitioning into a sol state upon heating and vice versa upon cooling. Notably, PHCS enabled media embedding, facilitating immunofluorescence histopathology. Additionally, the system demonstrated compatibility with various fluorescent probes, particularly lipophilic dyes. Our study successfully employed PHCS for the reconstruction of vascular structures within the intestine, enabling the generation of a 3D pathology model. These findings suggest that PHCS is a promising novel method for fabricating hydrogels for tissue clearing and holds great potential for application as a mounting medium for morphological imaging.

Stimulation of fracture mineralization by salt-inducible kinase inhibitors.

Over 6.8 million fractures occur annually in the US, with 10% experiencing delayed- or non-union. Anabolic therapeutics like PTH analogs stimulate fracture repair, and small molecule salt inducible kinase (SIK) inhibitors mimic PTH action. This study tests whether the SIK inhibitor YKL-05-099 accelerates fracture callus osteogenesis. 126 female mice underwent femoral shaft pinning and midshaft fracture, receiving daily injections of PBS, YKL-05-099, or PTH. Callus tissues were analyzed via RT-qPCR, histology, single-cell RNA-seq, and μCT imaging. Biomechanical testing evaluated tissue rigidity. A hydrogel-based delivery system for PTH and siRNAs targeting SIK2/SIK3 was developed and tested. YKL-05-099 and PTH-treated mice showed higher mineralized callus volume fraction and improved structural rigidity. RNA-seq indicated YKL-05-099 increased osteoblast subsets and reduced chondrocyte precursors. Hydrogel-released siRNAs maintained target knockdown, accelerating callus mineralization. YKL-05-099 enhances fracture repair, supporting selective SIK inhibitors' development for clinical use. Hydrogel-based siRNA delivery offers targeted localized treatment at fracture sites.

Cystic splenic lesions: a sonographic approach-retrospective study in 111 patients.

To describe and categorize splenic cystic-appearing lesions (S-CAL) with B-mode and contrast-enhanced ultrasound (CEUS). This retrospective study investigated S-CALs in 111 patients between 2003 and 2023 in an interdisciplinary ultrasound center with B-mode ultrasound, color Doppler sonography (CDS), and CEUS. S-CAL was characterized by echogenicity, diameter, and shape, as well as additional features like septation, calcification, or wall thickening, and CDS/CEUS were used to identify perfusion. Histological examination or imaging follow-up was necessary to determine the nature of S-CAL. Moreover, 'S-CAL with risk' was defined, necessitating further procedures. Four types (0-III) of S-CALs were defined based on ultrasound parameters. Fisher's exact test was used to compare non-parametric data. S-CAL of 111 patients (58 female, 53 men-average age: 58.6 years) was examined. Final diagnoses were: splenic cyst (n = 64, 57.7%); splenic abscess (n = 10, 9.0%); intrasplenic pseudoaneurysm (n = 10, 9.0%); splenic metastasis (n = 10, 9.0%); splenic infarction (n = 6, 5.4%); splenic hematoma (n = 4, 3.6%); other (n = 7, 6.3%). S-CAL groupings were type 0 (n = 11, 9.9%), type I (n = 33, 29.7%), type II (n = 24, 21.6%), and type III (n = 43, 38.7%). 'S-CAL with risk' was diagnosed in n = 41 (36.9%). Malignant S-CAL was only seen in type II (n = 2, 8.2%) and type III (n = 9, 20.9%) (p < 0.001). 'S-CALs with risk' were found more frequently in type 0 (n = 11, 100%), type II (n = 16, 66.7%) and type III (n = 13, 30.2%) than in type I (n = 1, 3%) (p < 0.001). B-mode ultrasound, CDS, and CEUS are useful to further characterize and follow-up S-CAL and identify 'S-CAL with risk', requiring further procedures. Ultrasound imaging is valuable for the detection, categorization, and monitoring of cystic-appearing lesions of the spleen, as well as for the identification of those with risk. An S-CAL may introduce uncertainty in clinical practice as imaging-based risk stratification is missing. B-mode and CEUS, along with the clinical context and follow-up, assist in characterizing and identifying 'S-CAL with risk'. S-CALs encompass various lesions, including simple cysts, metastases, abscesses, and intrasplenic pseudoaneurysms.

Focal point technology: Controlling treatment depth and pattern of skin injury by a novel highly focused laser

Selective photothermolysis has limitations in efficacy and safety for dermal targets. We describe a novel concept using scanned focused laser microbeams for precise control of dermal depth and pattern of injury, using a 1550nm laser that generates an array of conical thermal zones while minimizing injury to the epidermis. To characterize the conical thermal zones invivo and determine safe starting parameters to transition to a second phase to explore potential clinical indications. A focused toroidal (ring) laser beam was delivered through a cold sapphire window, sparing epidermal injury in a central zone. Pulse energy, lesion depth, density, and energy delivery were titrated in exvivo human skin and subsequently on the backs of 21 human subjects. Histology showed microscale patterns of thermal injury, which varied predictably with laser parameters. Time-course healing through histology and skin surface imaging demonstrated the ability of the device to deliver high energies without sequelae. Clinical data are currently being collected to further explore the safety and efficacy of the device. The 1550nm laser with focal point technology enables precise control of lesion depth while simultaneously sparing a large portion of the epidermis, lowering the risk of adverse effects.

Ouroboros: cross-linking protein expression perturbations and cancer histology imaging with generative-predictive modeling.

Imagine if we could simultaneously predict spatial protein expression in tissues from their routine Hematoxylin and Eosin (H&E) stained images, and create tissue images given protein expression profiles thus enabling virtual simulations of how protein expression alterations impact histology in complex diseases like cancer. Such an approach could lead to more informed diagnostic and therapeutic decisions for precision medicine at lower costs and shorter turnaround times, more detailed insights into underlying disease pathology as well as improvement in predictive and generative performance. In this study, we investigate the intricate correlation between protein expressions obtained from Hyperion mass cytometry and histopathological microstructures in conventional H&E stained glioblastoma (GBM) samples, unveiling morphological patterns and cellular-level spatial alterations associated with protein expression changes. To model these complex relationships, we propose a novel generative-predictive framework called Ouroboros for producing H&E images from protein expressions and simultaneously predicting protein expressions from H&E images. Our comprehensive sample-independent validation over 9920 tissue spots from 4 GBM samples encompassing visual image analysis, quantitative analysis, subspace alignment and perturbation experiments shows that the proposed generative-predictive approach offers significant improvements in predicting protein expression from images in comparison to baseline methods as well as accurate generation of virtual GBM sample images. This proof of concept study can contribute to advancing our understanding of histological responses to protein expression perturbations and lays the foundations for further developments in this area. Implementation and associated data for the proposed approach are available at the URL: https://github.com/Srijay/Ouroboros.

An efficient and cost-effective methods framework for performing biomechanical analysis on cadaveric skeletal muscle

IntroductionThe structure and arrangement of skeletal muscle fibers is the primary determinant of muscle function. While analysis of morphologic parameters to elucidate muscle structure-function relationships dates to the 17th century, considerable variability in methodology and technique to quantify these relationships exists within the literature. Additionally, applications to assess the impact of non-typical musculoskeletal morphologies on structure-function relationships are limited. This study aims to assemble and present a practical, step-by-step framework of combined methods and techniques for efficiently analyzing biomechanical impacts of cadaveric skeletal muscles. MethodsExisting skeletal muscle biomechanical formulas and experimentally determined parameters for typical fast-acting skeletal muscle were identified in the literature. The methods framework was assembled as a stepwise protocol that includes mathematical formulas, referenced accepted values, optional steps, section breaks, suggested techniques, important notes and footnotes, and materials needed for collecting necessary measurements. Proof of concept was achieved with primary histological data and imaging, along with references to examples where the methods have been successfully applied. ResultsThe assembled framework presents an ordered process for measuring skeletal muscle parameters, calculating the maximal isometric force of a skeletal muscle, and applying that data to understand musculoskeletal mechanics. Histology data, imaging, and a summary of studies that have successfully applied the framework methods provide visual aids and validation for the methods. ConclusionsThis study presents an efficient and convenient framework of combined methods and techniques for investigators to evaluate the biomechanical impact of skeletal muscles. The practical use of this framework should optimize project efficiency and spending, increase study rigor, and minimize procedural variation across different studies. This report may serve as a foundational resource for researchers studying cadaveric muscle biomechanics, and its use especially adds translational clinical value to case analyses of non-typical musculoskeletal morphologies.

Design of a clinical study to demonstrate the therapeutic benefit of another wound treatment product

Due to new legal requirements, a patient-relevant benefit for other wound treatment products (sPW) must be demonstrated through clinical studies if reimbursement at the expense of the statutory health insurance is sought in the non-inpatient sector. An interdisciplinary group with expertise in clinical wound care has developed general recommendations for the design of suitable studies. In addition to regulatory documents, previous studies that have already been recognized as proof of benefit in other areas served as a basis. Randomized controlled trials that cover at least the most common types of chronic wounds (arterial, venous, diabetic or pressure sore) are recommended as the best method for gathering evidence. Despite the heterogeneous etiology of chronic wounds, the results should also be transferable to other wound types. The test intervention does not usually consist of the sPW alone, but of a combined wound treatment that follows a treatment plan that is as clearly defined as possible. In the comparison group, all wound treatment options (besides the sPW) must also be available and used according to a similar predefined treatment plan. Depending on the intended purpose and treatment goal, complete wound closure should, if possible, be recorded as the cardinal - although not always as the primary - endpoint. In justified cases, e.g. in the case of intermediate use as part of phase-appropriate wound therapy for chronic wounds, a significant reduction in the wound area can also be considered for benefit assessment. Quality of life (e.g. pain) can also justify a benefit and can therefore be recorded as a primary outcome parameter in clinical trials. The duration of the clinical trial should be adapted to the central endpoints, the medical or nursing goal of wound care and the intended purpose of the sPW. A benefit does not always arise from microbiological, physiological, laboratory or histological parameters or imaging findings.