Diagnostic potential of blood plasma longitudinal viscosity measured using Brillouin light scattering.

Blood plasma viscosity (PV) is an established biomarker for numerous diseases. Measurement of the shear PV using conventional rheological techniques is, however, time consuming and requires significant plasma volumes. Here, we show that Brillouin light scattering (BLS) and angle-resolved spectroscopy measurements of the longitudinal PV from microliter-sized plasma volumes can serve as a proxy for the shear PV measured using conventional viscometers. This is not trivial given the distinct frequency regime probed and the longitudinal viscosity, a combination of the shear and bulk viscosity, representing a unique material property on account of the latter. We demonstrate this for plasma from healthy persons and patients suffering from different severities of COVID-19 (CoV), which has been associated with an increased shear PV. We further show that the additional information contained in the BLS-measured effective longitudinal PV and its temperature scaling can provide unique insight into the chemical constituents and physical properties of plasma that can be of diagnostic value. In particular, we find that changes in the effective longitudinal viscosity are consistent with an increased suspension concentration in CoV patient samples at elevated temperatures that is correlated with disease severity and progression. This is supported by results from rapid BLS spatial-mapping, angle-resolved BLS measurements, changes in the elastic scattering, and anomalies in the temperature scaling of the shear viscosity. Finally, we introduce a compact BLS probe to rapidly perform measurements in plastic transport tubes. Our results open a broad avenue for PV diagnostics based on the high-frequency effective longitudinal PV and show that BLS can provide a means for its implementation.

Arterio-venous Anastomoses of the Sucquet-Hoyer Type: Complexity and Distribution in the Human Dermis.

Our study aims at providing detailed information on numbers, form, and spatial distribution of arterio-venous anastomoses of the Sucquet-Hoyer type in the dermis of the nail bed, nail fold corner, thumb pad, arm, nose, glabella, lip, and ear. It further aims at providing a system, which relies on objective morphologic criteria for classifying Sucquet-Hoyer canals (SHCs). Using high-resolution episcopic microscopy (HREM), digital volume data of eight samples of each skin region were produced. Virtual three-dimensional (3D) models of the dermally located SHCs were created, and their 3D tortuosity (τ) values were determined. Dermal SHCs were identified in all 24 finger samples and in 1 lip sample. Beneath a field of 2 × 2 mm2, an average of four were located in the nail bed, three in the dermis of the thumb pad, and one in the dermis of the nail fold corner. Only a single dermal SHC was found in one lip sample. No SHCs were observed in the dermis of the other samples. The τ values of the SHCs ranged from 1.11 to 10. Building on these values, a classification system was designed, which distinguishes four SHC classes. The dermal distribution of the SHCs of different classes was similar in all specimens.

Successful evaluation of a new image-based parameter for the diagnosis of carpal tunnel syndrome: ultrasound assessment of longitudinal median nerve gliding in patients, healthy volunteers, and cadavers.

BACKGROUND: Reduced longitudinal median nerve gliding is a new promising diagnostic feature in carpal tunnel syndrome (CTS). However, the complexity of existing ultrasound analysis protocols undermines the application in routine clinical practice. AIM: To provide a simple method for assessing longitudinal gliding with ultrasound, without the need for post-hoc image analysis. DESIGN: 1) Retrospective cohort study, validation by external blinded reviewers; 2) proof of concept in body donors. SETTING: 1) Outpatient clinic; 2) anatomy department. POPULATION: The population included 48 patients with idiopathic CTS diagnosed by electrodiagnostic testing and ultrasound, as well as 15 healthy controls. Twelve, non-frozen, non-embalmed body donors were enrolled. METHODS: Longitudinal gliding of the median nerve in the carpal tunnel was visualized in all patients with idiopathic CTS and healthy controls. All ultrasound videos were pseudonymized, equipped with a scale, and randomized. Videos were analyzed by four independent radiologists, all blinded to clinical characteristics. The endpoint was gliding rated as millimeters. Validity of the technique was tested by using speckle tracking software, and in body donors, directly measuring nerve excursion in situ, simultaneously to ultrasound. RESULTS: Gliding differed significantly between controls and patients with CTS, decreasing with incremental CTS severity. A cut-off value of 3.5 mm to identify patients with CTS, yielded 93.8% sensitivity and 93.3% specificity. Intraclass correlation coefficient among senior author and raters was 0.798 (95% CI 0.513 to 0.900, P<0.001), indicating good reliability. Speckle tracking and especially direct validation in body donors correlated well with ultrasound findings. CONCLUSIONS: First, longitudinal median nerve gliding can reliably be assessed using this simple technique without the need for complicated procedures. Second, a decrease in gliding was found with progressive severity of CTS. Reproducibility for measured distances is good among raters. CLINICAL REHABILITATION IMPACT: An easy to apply sonography parameter would bolster the diagnostic ability of specialists in physical medicine and rehabilitation in daily routine.

Three-dimensional structural and metric characterisation of cardioids.

Exact three-dimensional (3D) structural information of developing organoids is key for optimising organoid generation and for studying experimental outcomes in organoid models. We set up a 3D imaging technique and studied complexly arranged native and experimentally challenged cardioids of two stages of remodelling. The imaging technique we employed is S-HREM (Scanning High Resolution Episcopic Microscopy), a variant of HREM, which captures multiple images of subsequently exposed surfaces of resin blocks and automatically combines them to large sized digital volume data of voxels sizes below 1 µm3. We provide precise volumetric information of the examined specimens and their single components and comparisons between stages in terms of volume and micro- and macroanatomic structure. We describe the 3D arrangement and lining of different types of cavities and their changes between day 10 and day 14 and map the various cell types to their precise spatial and structural environment. Exemplarily, we conducted semiautomatic counts of nuclei. In cryo-injured cardioids, we examined the extension and composition of the injured areas. Our results demonstrate the high quality and the great potential of digital volume data produced with S-HREM. It also provides sound metric and structural information, which assists production of native and experimentally challenged left ventricle cardioids and interpretation of their structural remodelling.

Effects of Tulp4 deficiency on murine embryonic development and adult phenotype.

Genetically engineered mouse models have the potential to unravel fundamental biological processes and provide mechanistic insights into the pathogenesis of human diseases. We have previously observed that germline genetic variation at the TULP4 locus influences clinical characteristics in patients with myeloproliferative neoplasms. To elucidate the role of TULP4 in pathological and physiological processes in vivo, we generated a Tulp4 knockout mouse model. Systemic Tulp4 deficiency exerted a strong impact on embryonic development in both Tulp4 homozygous null (Tulp4-/-) and heterozygous (Tulp4+/-) knockout mice, the former exhibiting perinatal lethality. High-resolution episcopic microscopy (HREM) of day 14.5 embryos allowed for the identification of multiple developmental defects in Tulp4-/- mice, including severe heart defects. Moreover, in Tulp4+/- embryos HREM revealed abnormalities of several organ systems, which per se do not affect prenatal or postnatal survival. In adult Tulp4+/- mice, extensive examinations of hematopoietic and cardiovascular features, involving histopathological surveys of multiple tissues as well as blood counts and immunophenotyping, did not provide evidence for anomalies as observed in corresponding embryos. Finally, evaluating a potential obesity-related phenotype as reported for other TULP family members revealed a trend for increased body weight of Tulp4+/- mice. RESEARCH HIGHLIGHTS: To study the role of the TULP4 gene in vivo, we generated a Tulp4 knockout mouse model. Correlative analyses involving HREM revealed a strong impact of Tulp4 deficiency on murine embryonic development.

Anatomical evaluation of a novel echocardiography based tricuspid valve classification in 60 hearts from body donors.

OBJECTIVES: This study aimed to provide comprehensive morphological descriptions of the morphology of the tricuspid valve and to evaluate if a novel echocardiography-based tricuspid valve nomenclature can also be understood anatomically. METHODS: Tricuspid valves of 60 non-embalmed human body donors without a medical history of pathologies or macroscopic malformations of the heart were included. Length, height and surface area of leaflets were measured. The valves were morphologically classified according to a novel echocardiography-based classification, in which 6 types are distinguished: classic 3-leaflet configuration, bicuspid valves, valves with 1 leaflet split into 2 scallops or leaflets and valves with 2 leaflets divided into 2 scallops or leaflets. RESULTS: We found a true 3-leaflet configuration in only 19 (31.7%) of valves. Five (8.3%) had a 2-leaflet configuration with a fused anterior and posterior leaflet. Of those, 3 had a divided septal leaflet. Four valves (6.7%) had a divided anterior leaflet, 17 (28.3%) had a divided posterior leaflet, 6 (10%) had a divided septal leaflet and 9 (15.0%) had 2 leaflets divided. Overall, 39 (65%) of valves have at least 1 leaflet that is divided. In 22 (36.7%) specimens, the leaflet was divided into true leaflets, and in 17 (28.3%) specimens, the leaflet was divided into scallops. In addition, we could identify 9 (15%) valves having 1 leaflet divided not only in 2 but 3 scallops or leaflets. CONCLUSIONS: This study provides further anatomical insight for the significant variability in the morphology of the tricuspid valve. By updating the understanding of its morphological characteristics, this study equips clinicians with valuable insights to effectively advance surgical and interventional treatment of tricuspid valves.

Revisiting the microsurgical anatomy of the sagittal stratum and surgical implications: fiber microdissection and tractography study.

OBJECTIVE: The term "sagittal stratum" was coined by Heinrich Sachs in 1892 to define a parasagittally oriented white matter layer at the temporo-occipital cortex. Although this term has been widely used for more than 100 years, the description, classification, borders, and involved fibers of the structure vary among authors and remain imprecise. Through fiber microdissection and tractography, the authors aimed to define the sagittal stratum and resolve the uncertainty by revealing the relationship of this structure to other cerebral white matter pathways and the orientation of fibers in it. METHODS: Twenty postmortem human cerebral hemispheres were prepared according to Klingler's method. Fiber dissections were performed under a surgical microscope and with microsurgical techniques. The results of dissection at each step were photographed with 2D and 3D imaging techniques, and 3D photogrammetry techniques were used to create a 360° model. Diffusion tensor imaging and 7T high-resolution MRI were used to confirm the findings. RESULTS: This study revisited the 3D organization of white matter tracts in the sagittal stratum through fiber microdissection and tractography. The microneuroanatomical structure of the sagittal stratum and its special organization with fibers from all three fiber systems are demonstrated. The authors' findings revealed that the sagittal stratum has two layers consisting of four different fiber tracts. Its external layer consists of a long association fiber and a commissural fiber, while its internal layer consists of intertwined projection fibers, including temporo-parieto-occipitopontine fibers and the posterior thalamic peduncle. Detailed microdissection also showed the location of the posterior thalamic peduncle in the most medial site of all posterior hemispheric projection fibers. CONCLUSIONS: The structure of the sagittal stratum is distinctive in that it contains all three main fiber systems: association, commissural, and projection. Because of its expansive location in the temporal and occipital lobes, it can be damaged by most neurosurgical pathologies and procedures. The authors emphasize the significance of preserving the sagittal stratum during surgical interventions while also challenging the notion of a "silent" brain, suggesting that the current inability to fully comprehend cerebral function contributes to this misconception. Detailed knowledge of the complex white matter anatomy of the sagittal stratum can guide neurosurgeons in surgical planning and the selection of appropriate surgical approaches with intraoperative orientation for safe surgery and less comorbidity.

Characterization, number, and spatial organization of nerve fibers in the human cervical vagus nerve and its superior cardiac branch.

BACKGROUND: Electrical stimulation of the vagus nerve (VN) is a therapy for epilepsy, obesity, depression, and heart diseases. However, whole nerve stimulation leads to side effects. We examined the neuroanatomy of the mid-cervical segment of the human VN and its superior cardiac branch to gain insight into the side effects of VN stimulation and aid in developing targeted stimulation strategies. METHODS: Nerve specimens were harvested from eight human body donors, then subjected to immunofluorescence and semiautomated quantification to determine the signature, quantity, and spatial distribution of different axonal categories. RESULTS: The right and left cervical VN (cVN) contained a total of 25,489 ± 2781 and 23,286 ± 3164 fibers, respectively. Two-thirds of the fibers were unmyelinated and one-third were myelinated. About three-quarters of the fibers in the right and left cVN were sensory (73.9 ± 7.5 % versus 72.4 ± 5.6 %), while 13.2 ± 1.8 % versus 13.3 ± 3.0 % were special visceromotor and parasympathetic, and 13 ± 5.9 % versus 14.3 ± 4.0 % were sympathetic. Special visceromotor and parasympathetic fibers formed clusters. The superior cardiac branches comprised parasympathetic, vagal sensory, and sympathetic fibers with the left cardiac branch containing more sympathetic fibers than the right (62.7 ± 5.4 % versus 19.8 ± 13.3 %), and 50 % of the left branch contained sensory and sympathetic fibers only. CONCLUSION: The study indicates that selective stimulation of vagal sensory and motor fibers is possible. However, it also highlights the potential risk of activating sympathetic fibers in the superior cardiac branch, especially on the left side.