Loss of Neuropilin2a/b or Sema3fa alters olfactory sensory axon dynamics and protoglomerular targeting

Background Olfactory Sensory Neuron (OSN) axons project from the zebrafish olfactory epithelium to reproducible intermediate target locations in the olfactory bulb called protoglomeruli at early stages in development. Two classes of OSNs expressing either OMP or TRPC2 exclusively target distinct, complementary protoglomeruli. Using RNAseq, we identified axon guidance receptors nrp2a and nrp2b, and their ligand sema3fa, as potential guidance factors that are differentially expressed between these two classes of OSNs. Methods To investigate their role in OSN axon guidance, we assessed the protoglomerular targeting fidelity of OSNs labeled by OMP:RFP and TRPC2:Venus transgenes in nrp2a, nrp2b, or sema3fa mutants. We used double mutant and genetic interaction experiments to interrogate the relationship between the three genes. We used live time-lapse imaging to compare the dynamic behaviors of OSN growth cones during protoglomerular targeting in heterozygous and mutant larvae. Results The fidelity of protoglomerular targeting of TRPC2-class OSNs is degraded in nrp2a, nrp2b, or sema3fa mutants, as axons misproject into OMP-specific protoglomeruli and other ectopic locations in the bulb. These misprojections are further enhanced in nrp2a;nrp2b double mutants suggesting that nrp2s work at least partially in parallel in the same guidance process. Results from genetic interaction experiments are consistent with sema3fa acting in the same biological pathway as both nrp2a and nrp2b. Live time-lapse imaging was used to examine the dynamic behavior of TRPC2-class growth cones in nrp2a mutants compared to heterozygous siblings. Some TRPC2-class growth cones ectopically enter the dorsal-medial region of the bulb in both groups, but in fully mutant embryos, they are less likely to correct the error through retraction. The same result was observed when TRPC2-class growth cone behavior was compared between sema3fa heterozygous and sema3fa mutant larvae. Conclusions Our results suggest that nrp2a and nrp2b expressed in TRPC2-class OSNs help prevent their mixing with axon projections in OMP-specific protoglomeruli, and further, that sema3fa helps to exclude TRPC2-class axons by repulsion from the dorsal-medial bulb.

Discrepancy in the Distribution Patterns of Subchondral Bone Density Across the Ankle Joint After Medial Opening-Wedge and Lateral Closing-Wedge High Tibial Osteotomy

Background: High tibial osteotomy (HTO) changes the alignment and dynamics of the ankle joint; however, differences in the stress distribution of the ankle joint after opening-wedge HTO (OWHTO) and closing-wedge HTO (CWHTO) are not understood. It is believed that subchondral bone density of the articular surface reflects the pattern of cumulative stress distribution across the joint surface. Purpose: To clarify the effects of OWHTO and CWHTO on the distribution patterns of subchondral bone density across the ankle joint using computed tomography (CT)–osteoabsorptiometry. Study Design: Cohort study; Level of evidence, 4. Methods: Radiographic and CT data of 18 cases who underwent OWHTO (OW group), 12 cases who underwent CWHTO (CW group), and 11 cases with unilateral anterior cruciate ligament injury serving as controls were retrospectively reviewed. The subchondral bone density of the distal tibia was assessed in the 3 groups using CT-osteoabsorptiometry. The distal tibial surface of the ankle joint was divided into 4 parts in the coronal direction, and the percentage of the high-density area (%HDA) to each subregion was compared before and after HTO. Results: Preoperatively, comparing %HDA among the 3 groups, there were no significant differences in any regions. In the OW group, postoperative %HDA in the most medial region was significantly increased compared with preoperative %HDA (49.3% to 53.0%; P = .011), and postoperative %HDA in the most lateral region was significantly decreased (21.4% to 17.2%; P = .003). On the other hand, in the CW group, postoperative %HDA in the most medial region was significantly decreased (55.7% to 35.7%; P = .001), and %HDA in the second lateral region was significantly increased (23.6% to 29.2%; P = .002). Conclusion: The ankle distribution pattern of subchondral bone density shifted significantly medially after OWHTO without fibular osteotomy, whereas the distribution pattern shifted laterally after CWHTO with fibular osteotomy. When the OWHTO is performed for patients with medial ankle osteoarthritis, surgeons should pay attention to potential postoperative progression of ankle osteoarthritis due to medial shift of the stress distribution in the ankle joint.

Micro-computed tomography analysis of the lumbar pedicle wall

Background Although the pedicle is routinely used as a surgical fixation site, the pedicle wall bone area fraction (bone area per unit area) and its distribution at the isthmus of the pedicle remain unknown. The bone area fraction at the pedicle isthmus is an important factor contributing to the strength of pedicle screw constructs. This study investigates the lumbar pedicle wall microstructure based on micro-computed tomography. Methods Six fresh-frozen cadaveric lumbar spines were analyzed. Left and right pedicles of each vertebra from L1 to L5 were resected for micro-computed tomography scanning. Data was analyzed with custom-written software to determine regional variation in pedicle wall bone area fraction. The pedicular cross-section was divided into four regions: lateral, medial, cranial, and caudal. The mean bone area fraction values for each region were calculated for all lumbar spine levels. Results The lateral region showed lower bone area fraction than the medial region at all spinal levels. Bone area fraction in the medial region was the highest at all levels except for L4, and the median values were 99.8% (95.9–100%). There were significant differences between the lateral region and the caudal region at L1, L2 and L3, but none at L4 and L5. The bone area fraction in the lateral region was less than 64% at all spinal levels and that in the caudal region was less than 67% at the L4 and L5 levels. Conclusions This study provides initial detailed data on the lumbar pedicle wall microstructure based on micro-computed tomography. These findings may explain why there is a higher incidence of pedicle screw breach in the pedicle lateral and caudal walls.

Characterization of Developmental Changes in Spontaneous Electrical Activity of Medial Superior Olivary Neurons Before Hearing Onset With a Combination of Injectable and Volatile Anesthesia

In this work the impact of two widely used anesthetics on the electrical activity of auditory brainstem neurons was studied during postnatal development. Spontaneous electrical activity in neonate rats of either sex was analyzed through a ventral craniotomy in mechanically ventilated pups to carry out patch clamp and multi-electrode electrophysiology recordings in the medial region of the superior olivary complex (SOC) between birth (postnatal day 0, P0) and P12. Recordings were obtained in pups anesthetized with the injectable mix of ketamine/xylazine (K/X mix), with the volatile anesthetic isoflurane (ISO), or in pups anesthetized with K/X mix that were also exposed to ISO. The results of patch clamp recordings demonstrate for the first time that olivary and periolivary neurons in the medial region of the SOC fire bursts of action potentials. The results of multielectrode recordings suggest that the firing pattern of single units recorded in K/X mix is similar to that recorded in ISO anesthetized rat pups. Taken together, the results of this study provide a framework to use injectable and volatile anesthetics for future studies to obtain functional information on the activity of medial superior olivary neurons in vivo.

Temperature gradients in domestic cats over seven-years-old: descriptive analysis

ABSTRACT: The temperature gradients are dynamic and noninvasive monitoring techniques that provide information on peripheral blood flow and have been related to the prognosis of patients with circulatory shock. This study evaluated 47 elderly domestic cats’ temperature gradients, and we measured central (rectal) and peripheral (palmar, plantar and medial region of the radio) temperatures. Values found in this study are compatible with studies in young felines and differ from dogs and humans. The mean gradients found were 7.5°C for the central-peripheral; 5.6°C for the peripheral-environmental; 2.7°C for the skin-diff; and 0°C for the member-diff and the variables age and gender do not seem to influence these measurements. To the authors’ knowledge, there is no description of temperature gradients in elderly domestic cats, so this study pretends to clarify the vasoconstriction response in this group of animals.

Partial rupture of the thigh adductor muscle in a dog: case report

ABSTRACT Muscle injuries are often reported in humans, but uncommon in dogs. The etiology is degenerative or traumatic, and traumatic is more common in athletes. The diagnosis is obtained by a combination of orthopedic physical examination and imaging diagnosis, such as ultrasonography. Therapy aims to optimize healing and avoid complications. The present study reports a case of partial rupture of thigh adductor muscle in a dog presenting acute lameness of the right pelvic limb. Swelling and pain in the medial region of the thigh was noticed. Ultrasound examination confirmed partial rupture of the thigh adductor muscle. The treatment included non-steroid anti-inflammatory, warm compresses, and rest. Ultrasound examinations were useful in this case to evaluate the progression of the lesion, which was rapid and satisfactory.

Multigene analysis of the catfish genus Trichomycterus and description of a new South American trichomycterine genus (Siluriformes, Trichomycteridae)

Trichomycterus comprises about 170 valid species, but its monophyly has been challenged in the last decades. Bayesian Inference and Maximum Likelihood analyses comprehending mitochondrial genes COI and CYTB and nuclear genes GLYT, MYH6 and RAG2 from 71 Trichomycterinae terminal taxa and eight outgroups were performed. The analyses highly supports a clade containing Trichomycterusnigricans, the type species of the genus, and several other congeners endemic to eastern and northeastern Brazil, herein considered as the genus Trichomycterus, the sister clade the southern Brazil and adjacent areas clade; the latter clade comprises two subclades, one comprising species of the genus Scleronema and another comprising species previously placed in Trichomycterus, herein described as a new genus. Cambevagen. n. is distinguished from all other trichomycterines by the presence of a bony flap on the channel of the maxillo-dentary ligament, the interopercle shorter than the opercle, a deep constriction on the basal portion of the antero-dorsal arm of the quadrate, absence of teeth in the coronoid process of the dentary, the maxilla shorter than the premaxilla, the cranial fontanel extending from the the medial posterior of frontal to the medial region of supraoccipital, and absence of the postorbital process of the sphenotic-prootic-pterosphenoid.

Biomechanical behaviour of a French femoral component with thin cement mantle

Objective Cement thickness of at least 2 mm is generally associated with more favorable results for the femoral component in cemented hip arthroplasty. However, French-designed stems have shown favorable outcomes even with thin cement mantle. The biomechanical behaviors of a French stem, Charnley-Marcel-Kerboull (CMK) and cement were researched in this study. Methods Six polished CMK stems were implanted into a composite femur, and one million times dynamic loading tests were performed. Stem subsidence and the compressive force at the bone-cement interface were measured. Tantalum ball (ball) migration in the cement was analyzed by micro CT Results The cement thickness of 95 % of the proximal and middle region was less than 2.5 mm. A small amount of stem subsidence was observed even with collar contact. The greatest compressive force was observed at the proximal medial region and significant positive correlation was observed between stem subsidence and compressive force. 9 of 11 balls in the medial region moved to the horizontal direction more than that of the perpendicular direction. The amount of ball movement distance in the perpendicular direction was 59 to 83% of the stem subsidence, which was thought to be slip in the cement of the stem. No cement defect and no cement breakage were seen. Conclusion Thin cement in CMK stems produced effective hoop stress without excessive stem and cement subsidence. Polished CMK stem may work like force-closed fixation in short-term experiment. Cite this article: Y. Numata, A. Kaneuji, L. Kerboull, E. Takahashi, T. Ichiseki, K. Fukui, J. Tsujioka, N. Kawahara. Biomechanical behaviour of a French femoral component with thin cement mantle: The ‘French paradox’ may not be a paradox after all. Bone Joint Res 2018;7:485–493. DOI: 10.1302/2046-3758.77.BJR-2017-0288.R2.

Wnt controls the medial–lateral subdivision of the Drosophila head

In insects, the subdivision of the head into a lateral region, harbouring the compound eyes (CEs), and a dorsal (medial) region, where the ocelli localize, is conserved. This organization might have been already present in the insects' euarthropodan ancestors. In Drosophila, the Wnt-1 homologue wingless (wg) plays a major role in the genetic subdivision of the head. To analyse specifically the role of wg signalling in the development of the dorsal head, we attenuated this pathway specifically in this region by genetic means. We find that loss of wg signalling transforms the dorsal/medial head into lateral head structures, including the development of ectopic CEs . Our genetic analysis further suggests that wg signalling organizes the dorsal head medial–lateral axis by controlling, at least in part, the expression domains of the transcription factors Otd and Ey/Pax6.

2010 Use it but still lose it: Exploring age-related changes in skeletal stem cell location and activation in response to physical stimulation

OBJECTIVES/SPECIFIC AIMS: Our goal is to assess age-related changes in osteogenic stem cell populations of bone tissue. We hypothesize that aging mice have reduced osteogenic capacity in response to physical stimulation due to aging-associated decline in osteoprogenitor cell number and their proliferative capacity. METHODS/STUDY POPULATION: Mechanical loading: The NYU School of Medicine Institutional Animal Care and Use Committee approved all procedures. The response of tibial periosteal cells to physical stimulation or mechanical loading was assessed in 16-week-old adult (n=6) and aged 78-week-old female (n=4) mice subjected to 4 consecutive days of strain-matched axial compressive loading (1400 μm, 120 cycles, 2 Hz). Whole Mount Staining: Baseline periosteal cell numbers and nuclear morphology were assessed by whole bone DAPI staining of the antero-medial region of the tibiae in adult and aged mice (n=6). Immunohistochemistry: Tibiae were fixed in 4% PFA, decalcified in 19% EDTA, OCT-embedded, and thickly sectioned (150 μm) at midshaft. Sca1+, Prrx1+, and Ki67+cell numbers were quantified by simultaneous fluorescent immunohistochemical staining from loaded and nonloaded contralateral tibiae. Nonimmune species specific serum served as negative controls. Imaging: 3D image datasets of the periosteum at the antero-medial region of the tibial midshaft were acquired by multi-photon and confocal microscopy. Quantification of Sca1+, Prrx1+, and Ki67+ cells was carried out using Particle Analysis software (ImageJ) and Imaris 7.4.2 Surface Rendering Statistics functions. Cell number was normalized to periosteal area (~0.04 mm2). A Student t-test determined significance at p<0.05. RESULTS/ANTICIPATED RESULTS: At baseline, aged periosteal cell nuclei (DAPI+) area (14% decrease, p<0.0001), nuclei number, and Prrx1+ cell number (22% decrease) was significantly lower compared with adult mice. In loaded adult mice, Prrx1+but not Sca1+cell number increased significantly (35%, p=0.0115). Proliferating Sca1+(top panel) and Prrx1+(top panel) cells also increased with loading, 62%, p=0.0253 and 115%, p=0.0004, respectively, in adult but not aged mice. The percentage of Prrx1+ cells undergoing proliferation (co-expressing Ki67+) in the total Prrx1+ cell population increased significantly with loading (bottom panel). Aged mice did not exhibit significant differences in loaded versus nonloaded controls for all other outcomes. Our data suggest fundamental changes in periosteal cell morphology, number and response to mechanical loading with aging. The significant increase in total Prrx1+ cell number and the number of Prrx1+ cells undergoing proliferation with loading in adult mice, suggest that the Prrx1+ cell population expands through proliferation. In fact, loading resulted in a 2-fold increase in the percentage of Prrx1+ preosteogenic cells undergoing proliferation. Accordingly, the significant age-related decrease in Prrx1+ cells may explain, in part, the attenuation of load-induced bone formation in aged mice. Loading resulted in greater numbers of proliferating Sca1+ cells (the more primitive cell) in adult mice, though this represented only a small percentage (<10%) of the total Sca1+ population. Mechanical loading expands the Prrx1+ pre-osteogenic cell population, but not the more primitive Sca1+ population. However, this load-induced osteogenic effect in the periosteum is not observed in aged mice, which may explain age-related diminishment of load-induced bone formation. DISCUSSION/SIGNIFICANCE OF IMPACT: Mechanical loading presents an inexpensive treatment for increasing bone mass and bone strength, but may be insufficient to prevent or reverse age-related bone loss due to reduced numbers of osteogenic progenitors in the periosteum. Therapeutic approaches targeting the osteogenic capacity of periosteal cells will be required to address declining mechanoresponsiveness with age.