Cone beam computed tomography and cross-sectional anatomy of the region of the fetlock in the horse (Equus caballus).

This study aimed to delineate the detailed anatomy of the metacarpophalangeal (MCP) and metatarsophalangeal (MTP) joints in healthy horses using cone beam computed tomography (CBCT). The fetlock region of 15 cadaveric forelimbs and 14 cadaveric hindlimbs from nine adult horses without orthopaedic disease underwent CBCT scanning. Additionally, arthrography CBCT scans were conducted following intra-articular injection of a radiopaque contrast medium containing blue epoxy resin dye. Subsequently, limbs were frozen and sectioned to visualize anatomical structures in sectional planes corresponding to selected CBCT images. CBCT proved suitable for detailed visualization of the bony components of the fetlock region. Furthermore, the common digital extensor tendon, superficial and deep digital flexor tendons, suspensory ligament, and straight and oblique sesamoidean ligaments were identifiable on CBCT images. However, certain ligaments, such as the collateral sesamoidean ligaments and intersesamoidean ligaments, were not clearly identified. The hyaline cartilage of the MCP and MTP joint facets was assessable on the post-contrast sequence. In cases where a radiographic or ultrasound examination cannot provide a definitive diagnosis and determine the extent of disease, CBCT can provide additional valuable data on the equine MCP and MTP joint. The images obtained in this study can serve as a reference for CBCT examination of the equine MCP and MTP joint.

3D Slicer open-source software plug-in for vector-based angle calculation of canine hind limb alignment in computed tomographic images.

BACKGROUND: Severe and complex angular limb deformities in dogs require accurate morphological assessment using diagnostic imaging to achieve successful orthopedic surgery. Computed tomography (CT) is commonly used to overcome projection errors in two-dimensional angular measurements of dog hindlimb alignment. Three-dimensional volume rendering (VR) techniques permit virtual positioning and variable projection, but the final CT-image that defines the projection plane for angular measurements remains two-dimensional. OBJECTIVE: We wanted to develop a true three-dimensional open-source technique to measure the alignments of the hind limbs of dogs in CT scanners. METHODS: We developed an open-source 3D Slicer plug-in, to perform angular measurements using vector calculations in three-dimensional space. In 113 CT-scans of canine pelvic limbs, femoral torsion, femoral varus, femorotibial rotation, tibial torsion, tibial varus and tibiotalar rotation angles were calculated and compared to an already validated technique using VoXim®. RESULTS: Reference points were identified and measurements were possible in the 113 acquisitions. The greatest difference between the two techniques was 1.4° at only one tibial torsion angle. Mean values for all Bland-Altman plots did not show significant differences and were less than 0.07° for all comparisons. DISCUSSION: Based on these results we considered angular measurements of canine hind limb alignment in CT scans using the 3D Slicer extension program sufficiently accurate for clinical orthopedic and surgical purposes in veterinary medicine. CONCLUSION: With our open-source 3D Slicer extension software, we provide a free accessible tool for veterinary orthopedic surgeons and thus we hope to improve angular measurements in CT-scans of canine hind limb deformities through true three-dimensionality.

Microvascular permeability and texture analysis of bone marrow in diabetic rabbits with critical limb ischemia based on dynamic contrast-enhanced magnetic resonance imaging.

BACKGROUND: Early on in the development of diabetes, skeletal muscles can exhibit microarchitectural changes that can be detected using texture analysis (TA) based on volume transfer constant (Ktrans) maps. Nevertheless, there have been few studies and thus we evaluated microvascular permeability and the TA of the bone marrow in diabetics with critical limb ischemia (CLI). METHODS: Eighteen male rabbits were randomly assigned equally into an operation group with hindlimb ischemia and diabetes, a sham-operated group with diabetes only, and a control group. Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) was performed on all rabbits at predetermined intervals (1, 5, 10, 15, 20, and 25 days post-surgery). The pharmacokinetic model was used to generate the permeability parameters, while the textural parameters were derived from the Ktrans map. Data analysis methods included the independent sample t-test, Mann-Whitney U test, repeated-measures analysis of variance, and Pearson correlation tests. RESULTS: The Ktrans values reached a minimum on day 1 after ischemia induction, then gradually recovered, but remained lower than those of the sham-operated group. The volume fraction only showed a significant difference between the operation group and the sham-operated group on day 5 post-surgery, but not in the extravascular extracellular space volume fraction at all time points. A significantly reduced Ktrans on day 1, a decreased number of bone trabeculae (Tb.N), and the area of bone trabeculae (Tb.Ar), and an increased microvessel density on day 25 in the operation group compared with the sham-operated group were observed. At each time point, there was a discernible difference between the two groups in the mean value, mean of positive pixels, and sumAverage. CONCLUSIONS: The early stages of diabetic bone marrow with CLI can be evaluated by DCE-MRI for microvascular permeability. Texture analysis based on DCE-MRI could act as an imaging discriminator and new radiological analysis tool for critical limb ischemia in diabetes mellitus.

Detecting medial patellar luxation with ensemble deep convolutional neural network based on a single rear view image of the hindlimb.

Medial patellar luxation (MPL) is a common orthopedic disease in dogs, which predisposes elderly and small-breed dogs. Unlike in humans, diagnosis in the early course of the disease is challenging because symptoms and joint-pain expression in canines are vague. Herein, we introduced a deep-learning system to diagnose MPL using a single rear-view hindlimb image. We believe that this is the first attempt to build a deep-learning system to diagnose MPL based on image analysis. Notably, 7689 images were collected from 2653 dogs in 30 private animal clinics between July 2021 and July 2022. Model performance was compared with ResNet50, VGG16, VGG19, Inception-V3, and veterinarian performance. For performance comparison, a professional veterinarian with > 10 years of experience selected images of 25 normal dogs and 25 dogs with MPL. The proposed model showed the highest performance, with 92.5% accuracy, whereas human experts showed an average accuracy of 55.2%. Therefore, our model can diagnose MPL using only a single rear-view hindlimb image. Furthermore, to solve the image uncertainty caused by the input image noise, we used a one-class SVM and ensemble learning methods to ensure model robustness. Our study will help diagnose MPL in clinical settings using a single rear-view hindlimb image.

Medial patellar luxation grade IV according to the Singleton system can be classified into skeletally immature and skeletally matured types.

OBJECTIVE: To determine the signalment and musculoskeletal morphology of small-breed dogs affected by medial patellar luxation (MPL) grade IV based on the age of the CT scan. ANIMALS: 40 small-breed dogs (54 limbs) with MPL grade IV. PROCEDURES: Dogs that had undergone corrective surgery for MPL grade IV and had performed CT of the hind limb before surgery were included. Signalment (age, body weight, sex, laterality, and breed) and concomitant cranial cruciate ligament rupture (CrCLR) were recorded. Femoral inclination angle, anatomical lateral distal femoral angle (aLDFA), femoral torsion angle, quadriceps muscle length to femoral length ratio (QML/FL), and patellar ligament length to patellar length were obtained by CT images. The dogs were categorized into 2 groups based on their age at the time of the CT scan, the skeletally immature group and the skeletally matured group. Signalment and group were included in the multiple regression analysis to determine the factors associated with each measurement parameter. A logistic regression analysis was conducted to determine the risk of CrCL concomitant with age. RESULTS: The multiple regression model demonstrated that the group was associated with the value of aLDFA and QML/FL. aLDFA was higher, and QML/FL was lower in group SI than in group SM. CrCLR was present in 5/54 limbs (9.2%), with a mean age of 70.8 months and it was associated with increasing age. CLINICAL RELEVANCE: In Singleton's classification, dogs classified as grade IV can be categorized into 2 groups based on musculoskeletal morphology and pathophysiology: the skeletally immature and skeletally matured types.

Comparison of Hindlimb Conformation in Cats with and without Medial Patellar Luxation.

OBJECTIVES: Medial patellar luxation (MPL) is the most common developmental cause of hindlimb lameness in cats. The association between femoral and tibial conformation and MPL measured on computed tomography (CT) has not been reported in cats. The aims were to report femoral and tibial conformation in cats with and without MPL and to report normal femoral and tibial angles. METHODS: Angle of inclination of femoral neck (AI), anatomical lateral distal femoral angle (aLDFA), femoral trochanteric angle (FCT), angle of anteversion of femoral neck (AA), distal and proximal anteversion angle (DAA/PAA), overall tibial valgus (TV), tibial torsion (TT), tibial tuberosity displacement (TTD) and trochlear depth:patellar thickness ratio (T:P) were measured by three observers on CT of cats with and without MPL. Comparisons were made between groups. Inter-observer intraclass correlation coefficient (ICC) was calculated. RESULTS: Sixteen cats were recruited: eight control and eight with MPL. The aLDFA, PAA, TT, TTD and T:P were significantly less in cats with high-grade MPL. The AI, FCT, AA, DAA and TV were not significantly different. A high correlation was shown with inter-observer ICC in 33.33% and good correlation in 26.67% when comparing measurements between observers. CLINICAL SIGNIFICANCE: This study suggests that cats with high-grade MPL have decreased TT, TTD and T:P and may require tibial tuberosity transposition and femoral trochleoplasty. The PAA, TT and aLDFA were decreased, although clinical significance may vary and these cats may not require correctional osteotomies. Results should be interpreted with caution as high/good levels of inter-observer ICC occurred in less than two-thirds of cases between observers.

Ultrasonographic Evaluation of the Suspensory Ligament in Quarter Horses Used for Cutting.

Ultrasound remains a mainstay in proximal suspensory ligament (PSL) evaluation. Despite recent improvements facilitating earlier diagnosis/treatment, needs exist for (1) characterization of normal ultrasonographic PSL cross-sectional area (CSA) in specific breeds/disciplines, (2) improved detection of early/subtle changes in the PSL, and (3) an understanding of suspensory ligament (SL) morphology change from work-related stress. The objectives of this study were to establish normal PSL CSA in Quarter Horses (QH) used for cutting via angle contrast ultrasonography, ultrasonographically monitor SL morphology as horses progress through training, and assess the impact of PSL disease on performance (Lifetime Earnings - $LTE). One hundred ten 2-year-old cutting horses had PSL ultrasound examination on all limbs, and examinations were repeated 2 years later on 51 remaining horses. PSL CSA at select locations was compared between time points and entire examinations subjectively graded by an equine radiologist. $LTE was obtained from the National Cutting Horse Association for each horse and analyzed by abnormality number, location, and development over time. A total of 267 PSL ultrasound examinations across both times points were measured and graded. Normal CSA was established by zone for forelimbs and hindlimbs. The prevalence of abnormal limbs increased between the 2-year-old and 4-year-old years, forelimb abnormalities were statistically more prevalent than hindlimb abnormalities at both time points, and osseous attachment abnormalities were statistically more prevalent than ligament abnormalities at both times points. No significant differences in PSL CSA change over time were seen in any group and there was no significant difference in $LTE among or between groups.

Anatomic, computed tomographic, and ultrasonographic assessment of the lymph nodes in presumed healthy adult cats: the abdomen, pelvis, and hindlimb.

BACKGROUND: The computed tomography (CT) and ultrasonography (US) features of lymph nodes of the abdomen, pelvis, and hindlimb in healthy cats are poorly described in the current literature. A prospective anatomic and reference interval study was therefore performed. The lymph nodes of six feline cadavers were identified, and dimensions were measured (length, width, and height). The lymph nodes from 30 healthy adult cats were identified and measured using CT (pre- and postcontrast) and US. The identification and dimensions of the separate lymph nodes were compared between imaging techniques and the anatomic study. RESULTS: The identification of lymph nodes was most frequent in CT, and the dimensions were overall larger than those identified and measured in US and the anatomic study. The caudal epigastric and sacral lymph nodes were not identified in the anatomic study. The ischiatic, lumbar aortic, internal iliac, and caudal epigastric lymph nodes were not visualized in US. The height presented the main statistical differences among techniques. The lymph nodes were mainly homogeneous in pre- and postcontrast CT and US images. Some lymph nodes showed a hyperattenuating periphery with a hypoattenuating center (on pre- and postcontrast images) and a hypo-/isoechoic periphery with a hyperechoic center, representing the hilar fat. The lymph nodes were commonly elongated and rounded except for the jejunal lymph nodes, which had an irregular shape. CONCLUSIONS: The assessment of most of the abdominal, pelvic, and hindlimb lymph nodes in the cat is feasible using CT and US, with CT performing best. Factors like the amount of adipose tissue and contrast administration subjectively improved the lymph node visualization and assessment. The measurements and features reported are proposed as reference values.

Microcomputed tomography versus plethysmometer and electronic caliper in the measurements of lymphedema in the hindlimb of mice.

Lymphedema affects 20% of women diagnosed with breast cancer. It is a pathology with no known cure. Animal models are essential to explore possible treatments to understand and potentially cure lymphedema. The rodent hindlimb lymphedema model is one of the most widely used. Different modalities have been used to measure lymphedema in the hindlimb of mice, and these are generally poorly assessed in terms of the interrater agreement; thus, there could be a risk of measuring bias and poor reproducibility. We examined the interrater agreement of µCT-scans, electronic caliper thickness of the paw and plethysmometer in the measurement of lymphedema in the hindlimb of mice. Three independent raters assessed 24 C57BL6 mice using these three modalities four times (week 1, 2, 4 and 8) with a total of 96 samples. The mean interrater differences were then calculated. The interrater agreement was highest in the µCT-scans, with an extremely low risk of measurement bias. The interrater agreement in the plethysmometer and electronic caliper was comparable with a low to moderate risk of measurement bias. The µCT-scanner should be used whenever possible. The electronic caliper should only be used if there is no µCT-scanner available. The plethysmometer should not be used in rodents of this size.

Computed tomography venography description of the normal anatomy of the canine hindlimb venous system.

This study was performed to provide a description of the normal anatomy of the canine hindlimb veins using helical CT images. The studies of 30 dogs that underwent CT venography with a 64-slice helical CT scanner were retrospectively reviewed. The dogs were positioned in a head-first prone or head-first supine body position. A topogram CT scan was performed from the pelvic limbs cranial to the iliac wings to the tips of the paws. Bolus-tracking software was used for the dual-phase angiogram, and contrast medium was administered in a cephalic vein. The venous phase was scanned after a delay time of a few seconds after the arterial phase in a caudocranial direction. Three-dimensional images were generated using maximum intensity projections and volume rendering technique. Representative images of three of the 30 dogs were selected and anatomic structures labelled.

High-Resolution Three-Dimensional Imaging of the Footpad Vasculature in a Murine Hindlimb Gangrene Model.

Peripheral arterial disease (PAD) is a significant cause of morbidity resulting from chronic exposure to atherosclerotic risk factors. Patients suffering from its most severe form, chronic limb-threatening ischemia (CLTI), face substantial impairments to daily living, including chronic pain, limited walking distance without pain, and nonhealing wounds. Preclinical models have been developed in various animals to study PAD, but mouse hindlimb ischemia remains the most widely used. There can be significant variation in response to ischemic insult in these models depending on the mouse strain used and the site, number, and means of arterial disruption. This protocol describes a unique method combining femoral artery and vein electrocoagulation with the administration of a nitric oxide synthase (NOS) inhibitor to reliably induce footpad gangrene in Friend Virus B (FVB) mice that resembles the tissue loss of CLTI. While traditional means of assessing reperfusion such as laser Doppler perfusion imaging (LDPI) are still recommended, intracardiac perfusion of the lipophilic dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) is used to label the vasculature. Subsequent whole-mount confocal laser scanning microscopy allows for high-resolution, three-dimensional (3D) reconstruction of footpad vascular networks that complements traditional means of assessing reperfusion in hindlimb ischemia models.

Contrast-Enhanced Ultrasound Reveals Partial Perfusion Recovery After Hindlimb Ischemia as Opposed to Full Recovery by Laser Doppler Perfusion Imaging.

Mouse models are critical in developing new therapeutic approaches to treat peripheral arterial disease (PAD). Despite decades of research and numerous clinical trials, the efficacy of available therapies is limited. This may suggest shortcomings in our current animal models and/or methods of assessment. We evaluated perfusion measurement methods in a mouse model of PAD by comparing laser Doppler perfusion imaging (LDPI, the most common technique), contrast-enhanced ultrasound (CEUS, an emerging technique) and fluorescent microspheres (conventional standard). Mice undergoing a femoral artery ligation were assessed by LDPI and CEUS at baseline and 1, 4, 7, 14, 28, 60, 90 and 150 d post-surgery to evaluate perfusion recovery in the ischemic hindlimb. Fourteen days after surgery, additional mice were measured with fluorescent microspheres, LDPI, and CEUS. LDPI and CEUS resulted in broadly similar trends of perfusion recovery until 7 d post-surgery. However, by day 14, LDPI indicated full recovery of perfusion, whereas CEUS indicated ∼50% recovery, which failed to improve even after 5 mo. In agreement with the CEUS results, fluorescent microspheres at day 14 post-surgery confirmed that perfusion recovery was incomplete. Histopathology and photoacoustic microscopy provided further evidence of sustained vascular abnormalities.

SPHARM-PDM based image preprocessing pipeline for quantitative morphometric analysis (QMA) for in situ joint assessment in rabbit and rat models.

The accessibility of quantitative measurements of joint morphometry depends on appropriate tibial alignment and volume of interest (VOI) selection of joint compartments; often a challenging and time-consuming manual task. In this work, we developed a novel automatic, efficient, and model-invariant image preprocessing pipeline that allows for highly reproducible 3D quantitative morphometric analysis (QMA) of the joint. The pipeline addresses the problem by deploying two modules: an alignment module and a subdivision module. Alignment is achieved by representing the tibia in its basic form using lower degree spherical harmonic basis functions and aligning using principal component analysis. The second module subdivides the joint into lateral and medial VOIs via a watershedding approach based on persistence homology. Multiple repeated micro-computed tomography scans of small (rat) and medium (rabbit) animal knees were processed using the pipeline to demonstrate model invariance. Existing QMA was performed to evaluate the pipeline's ability to generate reproducible measurements. Intraclass correlation coefficient and mean-normalised root-mean-squared error of more than 0.75 and lower than 9.5%, respectively, were achieved for joint centre of mass, joint contact area under virtual loading, joint space width, and joint space volume. Processing time and technical requirements were reduced compared to manual processing in previous studies.

Hind foot drumming: Volumetric micro-computed tomography investigation of the hind limb musculature of three African mole-rat species (Bathyergidae).

Several species of African mole-rats use seismic signalling by means of hind foot drumming for communication. The present study aimed to create three-dimensional reconstructions and compare volumetric measurements of 27 muscles of the hind limb of two drumming (Georychus capensis and Bathyergus suillus) and one non-drumming (Cryptomys hottentotus natalensis) species of African mole-rats. Diffusible iodine contrast-enhanced micro-computed tomography (diceCT) scans were performed on six specimens per species. Manual segmentation of the scans using VGMAX Studio imaging software allowed for individual muscles to be separated while automatically determining the volume of each muscle. The volume of the individual muscles was expressed as a percentage of the total hind limb volume and statistically compared between species. Subsequently, three-dimensional reconstructions of these muscles were created. Musculus gracilis anticus had a significantly larger percentage of the total hind limb muscle volume in both drumming species compared to the non-drumming C. h. natalensis. Furthermore, several hip and knee extensors, namely mm. gluteus superficialis, semimembranosus, gluteofemoralis, rectus femoris and vastus lateralis, had significantly larger muscle volume percentages in the two drumming species (G. capensis and B. suillus) compared to the non-drumming species. While not statistically significant, G. capensis had larger muscle volume percentages in several key hip and knee extensors compared to B. suillus. Additionally, G capensis had the largest summed percentage of the total hind limb volume in the hip flexor, hip extensor, knee extensor and ankle plantar flexor muscle groups in all the three species. This could be indicative of whole muscle hypertrophy in these muscles due to fast eccentric contractions that occur during hind foot drumming. However, significantly larger muscle volume percentages were observed in the scratch digging B. suillus compared to the other two chisel tooth digging species. Moreover, while not statistically significant, B. suillus had larger muscle volume percentages in several hip extensor and knee flexor muscles compared to G. capensis (except for m. vastus lateralis). These differences could be due to the large relative size of this species but could also be influenced by the scratch digging strategy employed by B. suillus. Therefore, while the action of hind foot drumming seems to influence certain key muscle volumes, digging strategy and body size may also play a role.

Sample Preparation for Computed Tomography-based Three-dimensional Visualization of Murine Hind-limb Vessels.

Blood vessels are complex networks with tree-like structures, and vascular networks are essential for maintaining both circulation and maintaining organ function. Clarifying the mechanism of blood vessel formation is therefore extremely useful for elucidating developmental processes and pathological mechanisms. Murine hind-limb vessels are often used as a model for physiological and pathological angiogenesis. Evaluation is mainly performed via a two-dimensional method using tissue sections. However, methods for evaluating three-dimensional (3D) vascular morphology are particularly limited. This paper introduces a method for visualizing murine hind-limbs using computed tomography (CT). Radiation-opaque resin is injected through the descending aorta, and whole vessels are filled with dye. By adjusting the time of dye injection, arterial-specific filling is also possible, and samples can be obtained with any micro-X-ray CT device. This contrast method provides a basic technique for the 3D evaluation of murine blood vessels in the lower extremities. Furthermore, this method can be used to visualize all blood vessels below the diaphragm and evaluate blood vessels in the abdominal organs.

Effects of Femorotibial and Tibiotarsal Joint Ranges of Motion on the Gastrocnemius Muscle-Tendon Working Length in the Canine Cadaveric Limb.

OBJECTIVE: This study aimed to describe changes in the length of the gastrocnemius muscle-tendon unit between the medial fabella and the calcaneus (MF-C) at different femorotibial joint (FTJ) and tibiotarsal joint (TTJ) angle combinations in the canine limb. STUDY DESIGN: This is an ex vivo kinematic study. ANIMALS: Canine cadaveric pelvic limbs (n = 11). PROCEDURES: One cadaveric pelvic limb was dissected to identify the anatomical origin and insertion of the gastrocnemius muscle-tendon unit for the radiographic landmark determination. The radiographic landmarks of the FTJ and TTJ were identified by lateral view radiographs. A series of lateral radiographs were evaluated to measure the linear MF-C at all combinations of the FTJ and TTJ angles. A mathematical model was defined for MF-C. RESULTS: The MF-C is associated with the FTJ and TTJ by a linear trigonometric relationship. Higher rate of increase in the MF-C was observed when the FTJ was > 135 degrees and the TTJ was < 125 degrees, compared with when the FTJ was <135 degrees and the TTJ was >125 degrees in all groups. CONCLUSIONS: Thirty degrees of free range of motion of the TTJ between 140 and 170 degrees did not elongate MF-C in this cadaveric limb study. CLINICAL RELEVANCE: Some degrees of free range of motion may be possible in the TTJ during tendon healing based on our cadaveric study without causing excessive change in the MF-C, although this concept should not be applied until isometric contractions of muscles are clearly understood. The relationship between the MF-C and tension at tenorrhaphy should be evaluated.

Zinc-containing Mohs' paste affects blood flow and angiogenesis suppression.

PURPOSE: Mohs' paste, which is composed of zinc chloride and zinc oxide starch, is used for hemostasis of superficial malignancy in the clinical setting. We investigated the concentration of intramuscular zinc in mice after Mohs' paste application and evaluated its relationship with angiogenesis from the perspective of blood flow levels within 24 h. METHODS: Male C57BL/6JJmsSlc mice were administered single dose of Mohs' paste at 25%, 50%, and 75% after unilateral hind limb surgery, and glycerin, a viscosity modifier, was administered to the control group (0%). Hind limb blood flow levels were measured with a laser Doppler perfusion imaging system (n = 6). The amounts of intramuscular zinc and vascular endothelial growth factor-A (VEGF-A) expression were analyzed using inductively coupled plasma mass spectrometry (ICP-MS) and western blotting, respectively (n = 5 or 3). RESULTS: Blood flow levels were significantly decreased in the 50% group after 8 h, and significantly decreased in the 25% and 50% groups after 24 h. Intramuscular zinc was significantly increased in the 50% and 75% groups after 8 h. Western blotting showed that VEGF-A levels were significantly increased in the 25% and 50% groups after 8 h. Based on analytical experiments and biological investigation, we predicated the pharmacological effect of Mohs' paste and found over 50% of it is critical in the blood flow and angiogenesis suppression after more than 8 h of its application. CONCLUSIONS: The results suggest that the mechanism of blood flow suppression is independent of VEGF-A levels and might suppress future angiogenesis. Our findings support that of previous studies, in which Mohs' paste was expected to induce hemostasis and suppress angiogenesis. It is an excellent ointment that facilitates hemostasis by suppressing blood flow regardless of angiogenesis, and may be apt for situations where hemostasis is required in the clinical setting.

Radiographical Survey of Osteochondrodysplasia in Scottish Fold Cats caused by the TRPV4 gene variant.

The unique appearance of Scottish Fold cats is caused by a single gene variant in TRPV4, which impacts the development of cartilage. This results in the ears folding forward and variable effects on articular cartilage and bone. While some find this appearance desirable, early work demonstrated that homozygous cats with two copies of this variant develop severe radiographic consequences. Subsequent breeding programs have mated heterozygous cats with straight-eared cats to ensure an equal mix of heterozygous (fold) and wild-type (nonfolded) offspring, in the hope of raising healthy cats. More recent radiological surveys suggest that these heterozygous cats may also have medical problems consisting of deformed distal extremities in the worst cases and accelerated onset of osteoarthritis. However, these previous studies were undermined by selection biases, lack of controls, unblinded assessment and lack of known genotypes. Our aim was to determine if heterozygous cats exhibit radiological abnormalities when controlling for these limitations. Specifically, DNA and radiographs were acquired for 22 Scottish Fold cats. Four reviewers, blinded to the ear phenotype, assessed the lateral radiographs. Genotyping showed that all 10 folded-ear cats were heterozygous, and none of the straight-ear cats (n = 12) had the abnormal TRPV4 variant. Although each reviewer, on average, gave a numerically worse 'severity score' to folded-ear cats relative to straight-ear cats, the images in heterozygous cats showed much milder radiological signs than previously published. This study provides additional information to be considered in the complicated debate as to whether cats with the TRPV4 variant should be bred for folded ears given the potential comorbidities.

Au-Doped Ag2 Te Quantum Dots with Bright NIR-IIb Fluorescence for In Situ Monitoring of Angiogenesis and Arteriogenesis in a Hindlimb Ischemic Model.

Fluorescence located in 1500-1700 nm (denoted as the near-infrared IIb window, NIR-IIb) has drawn great interest for bioimaging, owing to its ultrahigh tissue penetration depth and spatiotemporal resolution. Therefore, NIR-IIb fluorescent probes with high photoluminescence quantum yield (PLQY) and stability along with high biocompatibility are urgently pursued. Herein, a novel NIR-IIb fluorescent probe of Au-doped Ag2 Te (Au:Ag2 Te) quantum dots (QDs) is developed via a facile cation exchange method. The Au dopant concentration in the Ag2 Te QDs is tunable from 0% to 10% by controlling the ratio of supplied Au precursor to Ag2 Te QDs, resulting in a wide range of PL emission in the NIR-IIb window and a much-enhanced PL intensity. After surface modification, the Au:Ag2 Te QDs possess bright NIR-IIb emission, high colloidal stability and photostability, and decent biocompatibility. Further, in vivo monitoring of the process of angiogenesis and arteriogenesis in an ischemic hindlimb is successfully performed.

Laser Doppler Perfusion Imaging in the Mouse Hindlimb.

Blood flow recovery is a critical outcome measure after experimental hindlimb ischemia or ischemia-reperfusion. Laser Doppler perfusion imaging (LDPI) is a common, noninvasive, repeatable method for assessing blood flow recovery. The technique calculates overall blood flow in the sampled tissue from the Doppler shift in frequency caused when a laser hits moving red blood cells. Measurements are expressed in arbitrary perfusion units, so the contralateral non-intervened upon leg is usually used to help control measurements. Measurement depth is in the range of 0.3-1 mm; for hindlimb ischemia, this means that dermal perfusion is assessed. Dermal perfusion is dependent on several factors-most importantly skin temperature and anesthetic agent, which must be carefully controlled to result in reliable readings. Furthermore, hair and skin pigmentation can alter the ability of the laser to either reach or penetrate to the dermis. This article demonstrates the technique of LDPI in the mouse hindlimb.